A new plasmid display technology for the in vitro selection of functional phenotype–genotype linked proteins

Background: Display technologies which allow peptides or proteins to be physically associated with the encoding DNA are central to procedures which involve screening of protein libraries in vitro for new or altered function. Here we describe a new system designed specifically for the display of libraries of diverse, functional proteins which utilises the DNA binding protein nuclear factor κB (NF-κB) p50 to establish a phenotype–genotype link between the displayed protein and the encoding gene. Results: A range of model fusion proteins to either the amino- or carboxy-terminus of NF-κB p50 have been constructed and shown to retain the picomolar affinity and DNA specificity of wild-type NF-κB p50. Through use of an optimal combination of binding buffer and DNA target sequence, the half-life of p50–DNA complexes could be increased to over 47 h, enabling the competitive selection of a variety of protein–plasmid complexes with enrichment factors of up to 6000-fold per round. The p50-based plasmid display system was used to enrich a maltose binding protein complex to homogeneity in only three rounds from a binary mixture with a starting ratio of 1:108 and to enrich to near homogeneity a single functional protein from a phenotype–genotype linked Escherichia coli genomic library using in vitro functional selections. Conclusions: A new display technology is described which addresses the challenge of functional protein display. The results demonstrate that plasmid display is sufficiently sensitive to select a functional protein from large libraries and that it therefore represents a useful addition to the repertoire of display technologies.

Life cycle analysis for sustainability assessment of road projects

Road infrastructure has been considered as one of the most expensive and extensive infrastructure assets of the built environment globally. This asset also impacts the natural environment significantly during different phases of life e.g. construction, use, maintenance and end-of-life. The growing emphasis for sustainable development to meet the needs of future generations requires mitigation of the environmental impacts of road infrastructure during all phases of life e.g. construction, operation and end-of-life disposal (as required). Life-cycle analysis (LCA), a method of quantification of all stages of life, has recently been studied to explore all the environmental components of road projects due to limitations of generic environmental assessments. The LCA ensures collection and assessment of the inputs and outputs relating to any potential environmental factor of any system throughout its life. However, absence of a defined system boundary covering all potential environmental components restricts the findings of the current LCA studies. A review of the relevant published LCA studies has identified that environmental components such as rolling resistance of pavement, effect of solar radiation on pavement(albedo), traffic congestion during construction, and roadway lighting & signals are not considered by most of the studies. These components have potentially higher weightings for environment damage than several commonly considered components such as materials, transportation and equipment. This paper presents the findings of literature review, and suggests a system boundary model for LCA study of road infrastructure projects covering potential environmental components.

Validation of a parent-proxy quality of life questionnaire for paediatric chronic cough (PC-QOL)

Background Quality of life (QOL) measures are an important patient-relevant outcome measure for clinical studies. Currently there is no fully validated cough-specific QOL measure for paediatrics. The objective of this study was to validate a cough-specific QOL questionnaire for paediatric use. Method 43 children (28 males, 15 females; median age 29 months, IQR 20–41 months) newly referred for chronic cough participated. One parent of each child completed the 27-item Parent Cough-Specific QOL questionnaire (PC-QOL), and the generic child (Pediatric QOL Inventory 4.0 (PedsQL)) and parent QOL questionnaires (SF-12) and two cough-related measures (visual analogue score and verbal category descriptive score) on two occasions separated by 2–3 weeks. Cough counts were also objectively measured on both occasions. Results Internal consistency for both the domains and total PC-QOL at both test times was excellent (Cronbach alpha range 0.70–0.97). Evidence for repeatability and criterion validity was established, with significant correlations over time and significant relationships with the cough measures. The PC-QOL was sensitive to change across the test times and these changes were significantly related to changes in cough measures (PC-QOL with: verbal category descriptive score, rs=−0.37, p=0.016; visual analogue score, rs=−0.47, p=0.003). Significant correlations of the difference scores for the social domain of the PC-QOL and the domain and total scores of the PedsQL were also noted (rs=0.46, p=0.034). Conclusion The PC-QOL is a reliable and valid outcome measure that assesses QOL related to childhood cough at a given time point and measures changes in cough-specific QOL over time.

Feeling the life : a look into the visual culture of life scientists

In order to deal with human biological problems, life scientists have started investigating artificial ways of generating tissues and growing cells ? leading to the evolution of tissue engineering. In this paper we explore visualization practices of life scientists working within the domain of tissue engineering. We carried out a small scale ethnographic exploration with 8 scientists and explored that the real value of scientists' experiments (and simulations), reasoning and collaborative processes go beyond their end results. We observed that these scientists' three-dimensional reasoning, corporeal knowledge and intimacy with biological objects and tools play a vital role in overall success.

Women on the Heritage Register : the case of Nellie McCredie and Uanda House

Uanda house is of historical importance to Queensland both in terms of its architectural design and its social history. Uanda is a low set, single story house built in 1928, located in the inner city Brisbane suburb of Wilston. Architecturally, the house has a number of features that distinguish it from the surrounding bungalow influenced inter-war houses. The house has been described as a Queensland style house with neo-Georgian influences. Historically, it is associated with the entry of women into the profession of architecture in Queensland. Uanda is the only remaining intact work of architect/draftswoman Nellie McCredie and one of a very few examples of works by pioneering women architects in Queensland. The house was entered into the Queensland Heritage Register, in 2000, after an appeal against Brisbane City Council’s refusal of an application to demolish the house was disputed in the Queensland Planning and Environment court in 1998/1999. In the court’s report, Judge Robin QC, DCJ, stated that, “The importance of preserving women's history and heritage, often previously marginalised or lost, is now accepted at government level, recognising that role models are vital for bringing new generations of women into the professions and public life.” While acknowledging women’s contribution to the profession of architecture is an important endeavour, it also has the potential to isolate women architects as separate to a mainstream history of architecture. As Julie Willis writes, it can imply an atypical, feminine style of architecture. What is the impact or potential implications of recognising heritage buildings designed by women architects? The Judge also highlights the absence of a recorded history of unique Brisbane houses and questions the authority of the heritage register. This research looks at these points of difference through a case study of the Uanda house. The paper will investigate the processes of adding the house to the heritage register, the court case and existing research on Nellie McCredie and Uanda House.

A new plasmid display technology for the in vitro selection of functional phenotype-genotype linked proteins

Background Display technologies which allow peptides or proteins to be physically associated with the encoding DNA are central to procedures which involve screening of protein libraries in vitro for new or altered function. Here we describe a new system designed specifically for the display of libraries of diverse, functional proteins which utilises the DNA binding protein nuclear factor κB (NF-κB) p50 to establish a phenotype-genotype link between the displayed protein and the encoding gene. Results A range of model fusion proteins to either the amino- or carboxy-terminus of NF-κB p50 have been constructed and shown to retain the picomolar affinity and DNA specificity of wild-type NF-κB p50. Through use of an optimal combination of binding buffer and DNA target sequence, the half-life of p50-DNA complexes could be increased to over 47 h, enabling the competitive selection of a variety of protein-plasmid complexes with enrichment factors of up to 6000-fold per round. The p50-based plasmid display system was used to enrich a maltose binding protein complex to homogeneity in only three rounds from a binary mixture with a starting ratio of 1:108 and to enrich to near homogeneity a single functional protein from a phenotype-genotype linked Escherichia coli genomic library using in vitro functional selections. Conclusions A new display technology is described which addresses the challenge of functional protein display. The results demonstrate that plasmid display is sufficiently sensitive to select a functional protein from large libraries and that it therefore represents a useful addition to the repertoire of display technologies.

Practical Advice for Entrepreneurs

I am interested in the psychology of entrepreneurship—how entrepreneurs think, decide to act, and feel. I recently realized that while my publications in academic journals have implications for entrepreneurs, those implications have remained relatively hidden in the text of the articles and hidden in articles published in journals largely inaccessible to those involved in the entrepreneurial process. This book is designed to bring the practical implications of my research to the forefront. I decided to take a different approach with this book and not write it for a publisher. I did this because I wanted the ideas to be freely available: (1) I wanted those interested in practical advice for entrepreneurs to be able to freely download, distribute, and use this information (I only ask that the content be properly cited), (2) I wanted to release the chapters independently and make chapters available as they are finished, and; (3) I wanted this work to be a dialogue rather than a one-way conversation—I hope readers email me feedback (positive and negative) so that I can use this information to revise the book. In producing the journal articles underpinning this book, I have had the pleasure of working with many talented and wonderful colleagues—they are cited at the end of each chapter. I hope you find some of the advice in this book useful.

Teacher organizational citizenship behaviours and job efficacy : implications for student quality of school life

The present study investigated the impact of teachers' organizational citizenship behaviours (OCBs) on student quality of school life (SQSL) via the indirect effect of job efficacy. A measure of teacher OCBs was developed, tapping one dimension of individual-focused OCB (OCBI – student-directed behaviour) and two dimensions of organization-focused OCB (OCBO – civic virtue and professional development). In line with previous research suggesting that OCBs may enhance job efficacy, as well as studies demonstrating the positive effects of teacher efficacy on student outcomes, we expected an indirect relationship between teachers OCBs and SQSL via teachers' job efficacy. Hypotheses were tested in a multi-level design in which 170 teachers and their students (N=3,057) completed questionnaires. A significant proportion of variance in SQSL was attributable to classroom factors. Analyses revealed that the civic virtue and professional development behaviours of teachers were positively related to their job efficacy. The job efficacy of teachers also had a positive impact on all five indicators of SQSL. In regards to professional development, job efficacy acted as an indirect variable in the prediction of four student outcomes (i.e., general satisfaction, student–teacher relations, achievement, and opportunity) and fully mediated the direct negative effect on psychological distress.

Life transitions and changing physical activity patterns in young women

Background Physical activity (PA) patterns are likely to change in young adulthood in line with changes in lifestyle that occur in the transition from adolescence to adulthood. The aim of this study was to ascertain whether key life events experienced by young women in their early twenties are associated with increasing levels of inactivity. Methods This was a 4-year follow-up of 7281 participants (aged 18 to 23 years at baseline) in the Australian Longitudinal Study of Women’s Health, with self-reported measures of PA, life events, body mass index (BMI), and sociodemographic variables. Results The cross-sectional data indicated no change in PA between baseline (57% “active”) and follow-up (56% “active”). However, for almost 40% of the sample, PA category changed between baseline and follow-up, with approximately 20% of the women changing from being “active” to “inactive,” and another 20% changing from being “inactive” to “active.” After adjustment for age, other sociodemographic variables, BMI, and PA at baseline, women who reported getting married, having a first or subsequent child, or beginning paid work were more likely to be inactive at follow-up than those who did not report these events. Conclusions The results suggest that life events such as getting married, having children, and starting work are associated with decreased levels of PA in young adult women. Strategies are needed to promote maintenance of activity at the time when most women experience these key life-stage transitions.

Narratives of possession : reading for saga authorship

The aim of this thesis is to show how character analysis can be used to approach conceptions of saga authorship in medieval Iceland. The idea of possession is a metaphor that is adopted early in the thesis, and is used to describe Icelandic sagas as works in which traditional material is subtly interpreted by medieval authors. For example, we can say that if authors claim greater possession of the sagas, they interpret, and not merely record, the sagas' historical information. On the other hand, tradition holds onto its possession of the narrative whenever it is not possible for an author to develop his own creative and historical interests. The metaphor of possession also underpins the character analysis in the thesis, which is based on the idea that saga authors used characters as a vehicle by which to possess saga narratives and so develop their own historical interests. The idea of possession signals the kinds of problems of authorship study which are addressed here, in particular, the question of the authors' sense of saga writing as an act either of preservation or of creation. While, in that sense, the thesis represents an additional voice in a long-standing debate about the saga writers' relation to their source materials, I argue against a clear-cut distinction between creative and non-creative authors, and focus instead on the wide variation in authorial control over saga materials. This variation suggests that saga authorship is a multi-functional activity, or one which co-exists with tradition. Further, by emphasising characterisation as a method, I am adding to the weight of scholarship that seeks to understand the sagas in terms of their literary effects. The Introduction and chapter one lay out the theoretical scope of this thesis. My aim in these first two sections is to inform the reader of the type of critical questions that arise when authorship is approached in relation to characterisation, and to suggest an interpretive framework with which to approach these questions. In the Introduction this aim manifests as a brief discussion of the application of the term "authorship" to the medieval Icelandic corpus, a definition of the scope of this study, and an introduction to the connections, made throughout this thesis, between saga authors, the sagas' narrative style, and the style of characterisation in the sagas. Chapter one is a far more detailed discussion of our ability to make these connections. In particular, the chapter develops the definition of the analytical term "secondary authorship" that I introduce in order to delineate the type of characterisation that is of most interest in this thesis. "Secondary authorship" is a literary term that aims to sharpen our approach to saga authors' relationship to their characters by focusing on characters who make representations about the events of the saga. The term refers to any instance in which characters behave in a manner that resembles the creativity, interpretation, and understanding associated with authorship more generally. Character analysis cannot, however, be divorced from socio-historical approaches to the saga corpus. Most importantly, the sagas themselves are socio-historical representations that claim some degree of truth value. This claim that the sagas make by implication about their historicity is the starting point of a discussion of authorship in medieval Iceland. Therefore, at the beginning of chapter one I discuss some of the approaches to the social context of saga writing. This discussion serves as an introduction to both the culture of saga writing in medieval Iceland and to the nature of the sagas' historical perspective, and reflects my sense that literary interpretations of the sagas cannot be isolated from the historical discourses that frame them. The chapter also discusses possession, which, as I note above, is used alongside the concept of secondary authorship to describe the saga authors' relationship with the stories and characters of the past. At the close of chapter one, I offer a preliminary list the various functions of saga authorship, and give some examples of secondary authorship. From this point I am able to tie my argument about secondary authorship to specific examples from the sagas. Chapter two examines the effect of family obligations and domestic points of view in the depiction of characters' choices and conception of themselves. The examples that are given in that chapter - from Gisla saga Súrssonar and Íslendinga saga - are the first of a number of textual analyses that demonstrate the application of the concepts of secondary authorship and possession of saga narratives. The relationship between narratives about national and domestic matters shows how authorial creativity in the area of kinship obligation provides the basis for the saga's development of historical themes. Thus, the two major case studies given in chapter two tie authorial engagement with characters to the most influential social institution in early and medieval Iceland, the family. The remaining chapters represent similar attempts to relate authorial possession of saga characters to central socio-historical themes in the sagas, such as the settlement process in early Iceland and its influence on the development of regional political life (chapter three). Likewise, the strong authorial interest in an Icelander's journey to Norway in Heimskringla is presented as evidence of the author's use of a saga character to express an Icelandic interpretation of Norwegian history and to promote a sense that Iceland shared the ownership of regal history with Norway (chapter four). In that authorial engagement with the Icelander abroad, we witness saga characterisation being used as a basis for historical interpretation and the means by which foreign traditions and influence, not least the narratives of royal lives and of the Christianisation, are claimed as part of medieval Icelanders' self-conception. While saga authors observe the conventions of saga narration, characters are often subtly positioned as the authors' interpretive mirrors, especially clear than when they act as secondary authors. Nowhere is this more apparent than in Brennu- Njáls saga, which contains many characters who voice the author's claim to interpret the past. Even Hrútr Herjólfsson, through his remarkable perception of events and his conspicuous comments about them, acts as a secondary author by enabling the author to emphasise the importance of the disposition of characters. In Laxdœla saga and Þorgils saga ok Hafliða, authorial interest in characters' perception is matched by the thematising of learning, from the inception of knowledge as prophecy or advice to complete understanding by saga characters (chapter six). In Þorgils saga skarða, a character's inner development from an excessively ambitious and politically ruthless youth to a Christian leader killed by his kinsman allows the author to shape a political life into a lesson about leadership and the community's ability to moderate and contain the behaviour of extraordinary individuals. The portrayal draws on methods of characterisation that we can identify in Grettis saga Ásmundarson, Fóstbrœðra saga, and Orkneyinga saga. A comparison of the characterisation of figures with intense political or military ambitions suggests that saga authors were interested in the community's ability to balance their strength and ability with a degree of social moderation. The discussion of these sagas shows that character study can be used to analyse how the saga authors added their own voice to the voices passed down to medieval Icelanders in traditional narratives. Authorial engagement with characters allowed inherited traditions about early Norway and Iceland and records of thirteenth century events to be transformed into sophisticated historical works with highly creative elements. Through secondary authorship, saga authors took joint-possession of narratives and contested the power of tradition in setting the interpretive framework of a saga.

The importance of use and end-of-life phases to the life cycle greenhouse gas (GHG) emissions of concrete : a review

Global climate change is one of the most significant environmental impacts at the moment. One central issue for the building and construction industry to address global climate change is the development of credible carbon labelling schemes for building materials. Various carbon labelling schemes have been developed for concrete due to its high contribution to global greenhouse gas (GHG) emissions. However, as most carbon labelling schemes adopt cradle-to-gate as system boundary, the credibility of the eco-label information may not be satisfactory because recent studies show that the use and end-of-life phases can have a significant impact on the life cycle GHG emissions of concrete in terms of carbonation, maintenance and rehabilitation, other indirect emissions, and recycling activities. A comprehensive review on the life cycle assessment of concrete is presented to holistically examine the importance of use and end-of-life phases to the life cycle GHG quantification of concrete. The recent published ISO 14067: Carbon footprint of products – requirements and guidelines for quantification and communication also mandates the use of cradle-to-grave to provide publicly available eco-label information when the use and end-of-life phases of concrete can be appropriately simulated. With the support of Building Information Modelling (BIM) and other simulation technologies, the contribution of use and end-of-life phases to the life cycle GHG emissions of concrete should not be overlooked in future studies.

Discerning the timing and cause of historical mortality events in modern Porites from the Great Barrier Reef

The life history strategies of massive Porites corals make them a valuable resource not only as key providers of reef structure, but also as recorders of past environmental change. Yet recent documented evidence of an unprecedented increase in the frequency of mortality in Porites warrants investigation into the history of mortality and associated drivers. To achieve this, both an accurate chronology and an understanding of the life history strategies of Porites are necessary. Sixty-two individual Uranium–Thorium (U–Th) dates from 50 dead massive Porites colonies from the central inshore region of the Great Barrier Reef (GBR) revealed the timing of mortality to have occurred predominantly over two main periods from 1989.2 ± 4.1 to 2001.4 ± 4.1, and from 2006.4 ± 1.8 to 2008.4 ± 2.2 A.D., with a small number of colonies dating earlier. Overall, the peak ages of mortality are significantly correlated with maximum sea-surface temperature anomalies. Despite potential sampling bias, the frequency of mortality increased dramatically post-1980. These observations are similar to the results reported for the Southern South China Sea. High resolution measurements of Sr/Ca and Mg/Ca obtained from a well preserved sample that died in 1994.6 ± 2.3 revealed that the time of death occurred at the peak of sea surface temperatures (SST) during the austral summer. In contrast, Sr/Ca and Mg/Ca analysis in two colonies dated to 2006.9 ± 3.0 and 2008.3 ± 2.0, suggest that both died after the austral winter. An increase in Sr/Ca ratios and the presence of low Mg-calcite cements (as determined by SEM and elemental ratio analysis) in one of the colonies was attributed to stressful conditions that may have persisted for some time prior to mortality. For both colonies, however, the timing of mortality coincides with the 4th and 6th largest flood events reported for the Burdekin River in the past 60 years, implying that factors associated with terrestrial runoff may have been responsible for mortality. Our results show that a combination of U–Th and elemental ratio geochemistry can potentially be used to precisely and accurately determine the timing and season of mortality in modern massive Porites corals. For reefs where long-term monitoring data are absent, the ability to reconstruct historical events in coral communities may prove useful to reef managers by providing some baseline knowledge on disturbance history and associated drivers.