Evaluation of novel Streptococcus pyogenes vaccine candidates incorporating multiple conserved sequences from the C-repeat region of the M-protein

A major challenge for Streptococcus pyogenes vaccine development is the identification of epitopes that confer protection from infection by multiple S. pyogenes M-types. Here we have identified and characterised the distribution of common variant sequences from individual repeat units of the C-repeat region (CRR) of M-proteins representing 77 different M-types. Three polyvalent fusion vaccine candidates (SV1, SV2 and SV3) incorporating the most common variants were subsequently expressed and purified, and demonstrated to be alpha-helical by Circular Dichroism (CD), a secondary conformational characteristic of the CRR in the M-protein. Antibodies raised against each of these constructs recognise M-proteins that vary in their CRR, and bind to the surface of multiple S. pyogenes isolates. Antibodies raised against SV1, containing five variant sequences, also kill heterologous S. pyogenes isolates in in vitro bactericidal assays. Further structural characterisation of this construct demonstrated the conformation of SV1 was stable at different pHs, and thermal unfolding of SV1 a reversible process. Our findings demonstrate that linkage of multiple variant sequences into a single recombinant construct overcomes the need to embed the variant sequences in foreign helix promoting flanking sequences for conformational stability, and demonstrates the viability of the polyvalent candidates as global S. pyogenes vaccine candidates.

Performance testing of road bridge deck containing flat rail wagons

A road bridge containing disused flatbed rail wagons as the primary deck superstructure was performance tested in a low volume, high axle load traffic road in Queensland, Australia; some key results are presented in this paper. A fully laden truck of total weight 28.88 % of the serviceability design load prescribed in the Australian bridge code was used; its wheel positions were accurately captured using a high speed camera and synchronised with the real‐time deflections and strains measured at the critical members of the flat rail wagons. The strains remained well below the yield and narrated the existence of composite action between the reinforced concrete slab pavement and the wagon deck. A three dimensional grillage model was developed and calibrated using the test data, which established the structural adequacy of the rail wagons and the positive contribution of the reinforced concrete slab pavement to resist high axle traffic loads on a single lane bridge in the low volume roads network.

Smart Measures for Sustainable Outcomes: Integrated Considerations for Sustainable Refurbishment of Office Buildings

The increasing stock of aging office buildings will see a significant growth in retrofitting projects in Australian capital cities. Stakeholders of refitting works will also need to take on the sustainability challenge and realize tangible outcomes through project delivery. Traditionally, decision making for aged buildings, when facing the alternatives, is typically economically driven and on ad hoc basis. This leads to the tendency to either delay refitting for as long as possible thus causing building conditions to deteriorate, or simply demolish and rebuild with unjust financial burden. The technologies involved are often limited to typical strip-clean and repartition with dry walls and office cubicles. Changing business operational patterns, the efficiency of office space, and the demand on improved workplace environment, will need more innovative and intelligent approaches to refurbishing office buildings. For example, such projects may need to respond to political, social, environmental and financial implications. There is a need for the total consideration of buildings structural assessment, modeling of operating and maintenance costs, new architectural and engineering designs that maximise the utility of the existing structure and resulting productivity improvement, specific construction management procedures including procurement methods, work flow and scheduling and occupational health and safety. Recycling potential and conformance to codes may be other major issues. This paper introduces examples of Australian research projects which provided a more holistic approach to the decision making of refurbishing office space, using appropriate building technologies and products, assessment of residual service life, floor space optimisation and project procurement in order to bring about sustainable outcomes. The paper also discusses a specific case study on critical factors that influence key building components for these projects and issues for integrated decision support when dealing with the refurbishment, and indeed the “re-life”, of office buildings.

Greener and leaner : unleashing capacity of railroad concrete ties via limit states concept

New knowledge has raised a concern about the cost-ineffective design methods and the true performance of railroad prestressed concrete ties. Because of previous knowledge deficiencies, railway civil and track engineers have been aware of the conservative design methods for structural components in any railway track that rely on allowable stresses and material strength reductions. In particular, railway sleeper (or railroad tie) is an important component of railway tracks and is commonly made of prestressed concrete. The existing code for designing such components makes use of the permissible stress design concept, whereas the fiber stresses over cross sections at initial and final stages are limited by some empirical values. It is believed that the concrete ties complying with the permissible stress concept possess unduly untapped fracture toughness, based on a number of proven experiments and field data. Collaborative research run by the Australian Cooperative Research Centre for Railway Engineering and Technologies (Rail CRC) was initiated to ascertain the reserved capacity of Australian railway prestressed concrete ties that were designed using the existing design code. The findings have led to the development of a new limit-states design concept. This paper highlights the conventional and the new limit-states design philosophies and their implication to both the railway community and the public. © 2011 American Society of Civil Engineers.

Investing in the future : analysis of TxDOT 1547 Over-Axle Weight Tolerance Permits

The trucking industry has played a significant role in the economic growth in Texas by transporting and distributing commodities using commercial motor vehicles. The Texas Department of Transportation (TxDOT), however, has recognized that the large number of overweight trucks operating on the state highway system has resulted in the deterioration of pavement condition. In addition, the permit fee to carry higher loads above legal limits is much lower than the cost to treat the increase in pavement damage. The primary purpose of the research presented in this paper is to investigate current TxDOT overweight permit structures to support pavement management. The research team analyzed the TxDOT “1547” Over-axle Weight Tolerance Permit structure to support an increase in the fee structure, bringing it more in line with the actual pavement damage. The analysis showed that the revised overweight permit structure could provide an additional $9.3 million annually for pavement maintenance needs by increasing current permit fees. These results were supported by the 2030 Committee for recommendation to the Texas Transportation Commission and consideration by the State Legislature [1]. The research team recommends conducting further research to identify methods for working cooperatively with the trucking industry to develop improved methods for assessing weight damage relationships and developing more effective and accurate means for assessing overweight permit fees.

Young adults’ accounts of scientific knowledge when responding to a television news report of contested science

This paper reports one aspect of a study of 28 young adults (18–26 years) engaging with the uncertain (contested) science of a television news report about recent research into mobile phone health risks. The aim of the study was to examine these young people’s ‘accounts of scientific knowledge’ in this context. Seven groups of friends responded to the news report, initially in focus group discussions. Later in semi-structured interviews they elaborated their understanding of the nature of science through their explanations of the scientists’ disagreement and described their mobile phone safety risk assessments. This paper presents their accounts in terms of their views of the nature of science and their concept understanding. Discussions were audio-recorded then analysed by coding the talk in terms of issues raised, which were grouped into themes and interpreted in terms of a moderate social constructionist theoretical framing. In this context, most participants expressed a ‘common sense’ view of the nature of science, describing it as an atheoretical, technical procedure of scientists testing their personal opinions on the issue, subject to the influence of funding sponsors. The roles of theory and data interpretation were largely ignored. It is argued that the nature of science understanding is crucial to engagement with contemporary socioscientific issues, particularly the roles of argumentation, theory, data interpretation, and the distinction of science from common sense. Implications for school science relate primarily to nature of science teaching and the inclusion of socioscientific issues in school science curricula. Future research directions are considered.

China and the "Responsibility to Protect" (R2P) : the implications of the Libyan intervention

The emerging principle of a “responsibility to protect” (R2P) presents a direct challenge to China's traditional emphasis on the twin principles of non-intervention in the domestic affairs of other states and non-use of military force. This paper considers the evolution of China‟s relationship with R2P over the past ten years. In particular, it examines how China engaged with R2P during the recent Libyan crisis, and considers what impact this conflict may have first, on Chinese attitudes to R2P, and second, on the future development and implementation of the doctrine itself. This paper argues that China‟s decision to allow the passage of Security Council resolution 1973, authorising force in Libya, was shaped by an unusual set of pragmatic considerations, and should not be viewed as evidence of a dramatic shift in Chinese attitudes towards R2P. More broadly, controversy over the scope of NATO's military action in Libya has raised questions about R2P‟s legitimacy, which has contributed to a lack of timely international action in Syria and Yemen. In the short term at least, this post-Libya backlash against R2P is likely to constrain the Security Council‟s ability to respond decisively to other civilian protection situations.

Defining the role of phytoene synthase in carotenoid accumulation of high provitamin A bananas

Vitamin A deficiency (VAD) is a serious problem in developing countries, affecting approximately 127 million children of preschool age and 7.2 million pregnant women each year. However, this deficiency is readily treated and prevented through adequate nutrition. This can potentially be achieved through genetically engineered biofortification of staple food crops to enhance provitamin A (pVA) carotenoid content. Bananas are the fourth most important food crop with an annual production of 100 million tonnes and are widely consumed in areas affected by VAD. However, the fruit pVA content of most widely consumed banana cultivars is low (~ 0.2 to 0.5 ìg/g dry weight). This includes cultivars such as the East African highland banana (EAHB), the staple crop in countries such as Uganda, where annual banana consumption is approximately 250 kg per person. This fact, in addition to the agronomic properties of staple banana cultivars such as vegetative reproduction and continuous cropping, make bananas an ideal target for pVA enhancement through genetic engineering. Interestingly, there are banana varieties known with high fruit pVA content (up to 27.8 ìg/g dry weight), although they are not widely consumed due to factors such as cultural preference and availability. The genes involved in carotenoid accumulation during banana fruit ripening have not been well studied and an understanding of the molecular basis for the differential capacity of bananas to accumulate carotenoids may impact on the effective production of genetically engineered high pVA bananas. The production of phytoene by the enzyme phytoene synthase (PSY) has been shown to be an important rate limiting determinant of pVA accumulation in crop systems such as maize and rice. Manipulation of this gene in rice has been used successfully to produce Golden Rice, which exhibits higher seed endosperm pVA levels than wild type plants. Therefore, it was hypothesised that differences between high and low pVA accumulating bananas could be due either to differences in PSY enzyme activity or factors regulating the expression of the psy gene. Therefore, the aim of this thesis was to investigate the role of PSY in accumulation of pVA in banana fruit of representative high (Asupina) and low (Cavendish) pVA banana cultivars by comparing the nucleic acid and encoded amino acid sequences of the banana psy genes, in vivo enzyme activity of PSY in rice callus and expression of PSY through analysis of promoter activity and mRNA levels. Initially, partial sequences of the psy coding region from five banana cultivars were obtained using reverse transcriptase (RT)-PCR with degenerate primers designed to conserved amino acids in the coding region of available psy sequences from other plants. Based on phylogenetic analysis and comparison to maize psy sequences, it was found that in banana, psy occurs as a gene family of at least three members (psy1, psy2a and psy2b). Subsequent analysis of the complete coding regions of these genes from Asupina and Cavendish suggested that they were all capable of producing functional proteins due to high conservation in the catalytic domain. However, inability to obtain the complete mRNA sequences of Cavendish psy2a, and isolation of two non-functional Cavendish psy2a coding region variants, suggested that psy2a expression may be impaired in Cavendish. Sequence analysis indicated that these Cavendish psy2a coding region variants may have resulted from alternate splicing. Evidence of alternate splicing was also observed in one Asupina psy1 coding region variant, which was predicted to produce a functional PSY1 isoform. The complete mRNA sequence of the psy2b coding regions could not be isolated from either cultivar. Interestingly, psy1 was cloned predominantly from leaf while psy2 was obtained preferentially from fruit, suggesting some level of tissue-specific expression. The Asupina and Cavendish psy1 and psy2a coding regions were subsequently expressed in rice callus and the activity of the enzymes compared in vivo through visual observation and quantitative measurement of carotenoid accumulation. The maize B73 psy1 coding region was included as a positive control. After several weeks on selection, regenerating calli showed a range of colours from white to dark orange representing various levels of carotenoid accumulation. These results confirmed that the banana psy coding regions were all capable of producing functional enzymes. No statistically significant differences in levels of activity were observed between banana PSYs, suggesting that differences in PSY activity were not responsible for differences in the fruit pVA content of Asupina and Cavendish. The psy1 and psy2a promoter sequences were isolated from Asupina and Cavendish gDNA using a PCR-based genome walking strategy. Interestingly, three Cavendish psy2a promoter clones of different sizes, representing possible allelic variants, were identified while only single promoter sequences were obtained for the other Asupina and Cavendish psy genes. Bioinformatic analysis of these sequences identified motifs that were previously characterised in the Arabidopsis psy promoter. Notably, an ATCTA motif associated with basal expression in Arabidopsis was identified in all promoters with the exception of two of the Cavendish psy2a promoter clones (Cpsy2apr2 and Cpsy2apr3). G1 and G2 motifs, linked to light-regulated responses in Arabidopsis, appeared to be differentially distributed between psy1 and psy2a promoters. In the untranscribed regulatory regions, the G1 motifs were found only in psy1 promoters, while the G2 motifs were found only in psy2a. Interestingly, both ATCTA and G2 motifs were identified in the 5’ UTRs of Asupina and Cavendish psy1. Consistent with other monocot promoters, introns were present in the Asupina and Cavendish psy1 5’ UTRs, while none were observed in the psy2a 5’ UTRs. Promoters were cloned into expression constructs, driving the â-glucuronidase (GUS) reporter gene. Transient expression of the Asupina and Cavendish psy1 and psy2a promoters in both Cavendish embryogenic cells and Cavendish fruit demonstrated that all promoters were active, except Cpsy2apr2 and Cpsy2apr3. The functional Cavendish psy2a promoter (Cpsy2apr1) appeared to have activity similar to the Asupina psy2a promoter. The activities of the Asupina and Cavendish psy1 promoters were similar to each other, and comparable to those of the functional psy2a promoters. Semi-quantitative PCR analysis of Asupina and Cavendish psy1 and psy2a transcripts showed that psy2a levels were high in green fruit and decreased during ripening, reinforcing the hypothesis that fruit pVA levels were largely dependent on levels of psy2a expression. Additionally, semi-quantitative PCR using intron-spanning primers indicated that high levels of unprocessed psy2a and psy2b mRNA were present in the ripe fruit of Cavendish but not in Asupina. This raised the possibility that differences in intron processing may influence pVA accumulation in Asupina and Cavendish. In this study the role of PSY in banana pVA accumulation was analysed at a number of different levels. Both mRNA accumulation and promoter activity of psy genes studied were very similar between Asupina and Cavendish. However, in several experiments there was evidence of cryptic or alternate splicing that differed in Cavendish compared to Asupina, although these differences were not conclusively linked to the differences in fruit pVA accumulation between Asupina and Cavendish. Therefore, other carotenoid biosynthetic genes or regulatory mechanisms may be involved in determining pVA levels in these cultivars. This study has contributed to an increased understanding of the role of PSY in the production of pVA carotenoids in banana fruit, corroborating the importance of this enzyme in regulating carotenoid production. Ultimately, this work may serve to inform future research into pVA accumulation in important crop varieties such as the EAHB and the discovery of avenues to improve such crops through genetic modification.

US Federal Government entrepreneurship : new enterprise structures

Purpose: The purpose of this paper is to guide the formation and to determine the structure of new governmental entrepreneurial ventures based on the nature of the public goods and the need for an entrepreneurial orientation. Design/methodology/research: This paper is conceptual and is based on reviews of appropriate organizational structure and entrepreneurship, cases studies, and the authors' experiences. Findings: Public or quasi-public entities may need to change their organizational structure in order to act more entrepreneurially and to be more effective in accomplishing their missions. Propositions are raised to guide the development of new public or private enterprises and provide the basis for future research. Research limitations/implications: This paper is conceptual and needs to be tested empirically. Though other levels of government and countries were included, a major focus is on the US federal government. Originality/value: This is the first published research on the topic of new enterprise government structures based on the nature of the goods and the requisite entrepreneurial orientation. It will help governmental and quasi-governmental organizations in developing efficient and effective organizational structures.

Integrating complementary and alternative therapies into psychological practice : A qualitative analysis

Although complementary and alternative therapies (CATs) are utilised widely for treating psychological disorders, little research has examined psychologists' beliefs about integrating CAT into psychological practice. Six practicing psychologists and six psychology students were interviewed about their CAT integration beliefs, in particular integrating CAT into clinical practice via recommending CATs, offering referrals to CAT practitioners, or undertaking training to utilise CATs within psychological practice. Guided broadly from a theory of planned behaviour perspective, participants raised a number of costs and benefits, discussed referent groups who would influence their decisions, and suggested motivators and barriers for integration. A number of additional themes were raised, including risks, such as the possibility of litigation and the need for clear Society guidelines, as most participants were unclear about what constitutes appropriate practice. Identifying these themes serves as an important initial step to informing discussion and policy for this emerging practice issue within psychology.

Move it, use it : university-school collaboration for learning to teach physical education in primary schools

Reports of increasing numbers of obese Australian children and adolescents have raised the alarm to be proactive in reducing this so called epidemic. It has evoked a call for greater emphasis on teaching physical education in schools, as a measure for attaining fitness not only with obese students but for all students. This paper emphasises how preservice teachers need to be a key target for implementing physical education (PE) reform in schools, as many primary teachers will be generalists and may not be confident enough to implement PE effectively. Through a review of existing literature, teaching practices essential for the effective promotion and implementation of PE were identified under six broad categories: personal-professional skills development, addressing system requirements, pedagogical practices, managing student behaviour, providing feedback to students, and reflecting on practice. Subsequently, the development of these practices in preservice teachers is considered in the context of a university-school collaboration where preservice teachers taught physical education to primary school students for one day per week over a four week period. These authentic teaching experiences provided the preservice teachers with vital opportunities to put theory into practice and interact with “real-world” students. Self-evaluative data from 38 of these preservice teachers, in the form of a five-part Likert scale survey and extended response survey, demonstrated that they were able to develop the majority of the essential teaching practices identified by literature. In particular, the preservice teachers developed self efficacy, enthusiasm, and motivation for teaching PE, facets which are often found to be lacking in generalist primary teachers and yet are essential if children’s perceptions and habits regarding physical activity are to be changed.

Perceptions of visibility and conspicuity of biomotion clothing configurations for road workers at road work sites

Aims: This study determined whether the visibility benefits of positioning retroreflective strips in biological motion configurations were evident at real world road worker sites. Methods: 20 visually normal drivers (M=40.3 years) participated in this study that was conducted at two road work sites (one suburban and one freeway) on two separate nights. At each site, four road workers walked in place wearing one of four different clothing options: a) standard road worker night vest, b) standard night vest plus retroreflective strips on thighs, c) standard night vest plus retroreflective strips on ankles and knees, d) standard night vest plus retroreflective strips on eight moveable joints (full biomotion). Participants seated in stationary vehicles at three different distances (80m, 160m, 240m) rated the relative conspicuity of the four road workers using a series of a standardized visibility and ranking scales. Results: Adding retroreflective strips in the full biomotion configuration to the standard night vest significantly (p<0.001) enhanced perceptions of road worker visibility compared to the standard vest alone, or in combination with thigh retroreflective markings. These visibility benefits were evident at all distances and at both sites. Retroreflective markings at the ankles and knees also provided visibility benefits compared to the standard vest, however, the full biomotion configuration was significantly better than all of the other configurations. Conclusions: These data provide the first evidence that the benefits of biomotion retroreflective markings that have been previously demonstrated under laboratory and closed- and open-road conditions are also evident at real work sites.

Communicating with the coroner : how religion, culture, and family concerns may influence autopsy decision making

Based on coronial data gathered in the state of Queensland in 2004, this article reviews how a change in legislation may have impacted autopsy decision making by coroners. More specifically, the authors evaluated whether the requirement that coronial autopsy orders specify the level of invasiveness of an autopsy to be performed by a pathologist was affected by the further requirement that coroners take into consideration a known religion, culture, and/or raised family concern before making such an order. Preliminary data reveal that the cultural status of the deceased did not affect coronial autopsy decision making. However, a known religion with a proscription against autopsy and a raised family concern appeared to be taken into account by coroners when making autopsy decisions and tended to decrease the invasiveness of the autopsy ordered from a full internal examination to either a partial internal examination or an external-only examination of the body. The impact of these findings is briefly discussed.

‘Show Me Your Wiki and I’ll Show you Mine’: Using Online Interactive Media to Improve Academic Writing and Research in a Public Health Under-Graduate Cohort

The number of Internet users in Australia has been steadily increasing, with over 10.9 million people currently subscribed to an internet provider (ABS, 2011). Over the past year, the most avid users of the Internet were 15 – 24 year olds, with approximately 95% accessing the internet on a regular basis (ABS, Social Trends, 2011). While the internet has been described as fundamental to higher education students, social and leisure internet tools are also increasingly being used by these students to generate and maintain their social and professional networks and interactions (Duffy & Bruns 2006). Rapid technological advancements have enabled greater and faster access to information for learning and education (Hemmi et al, 2009; Glassman and Kang, 2011). As such, we sought to integrate interactive, online social media into the assessment profile of a Public Health undergraduate cohort at the Queensland University of Technology (QUT). The aim of this exercise was to engage students to both develop and showcase their research on a range of complex, contemporary health issues within the online forum of Wikispaces (http://www.wikispaces.com/) for review and critique by their peers. We applied Bandura’s Social Learning Theory (SLT) to analyse the interactive processes from which students developed deeper and more sustained learning, and via which their overall academic writing standards were raised. This paper outlines the assessment task, and the students’ feedback on their learning outcomes in relation to the Attentional, Retentional, Motor Reproduction, and Motivational Processes outlined by Bandura in SLT. We conceptualise the findings in a theoretical model, and discuss the implications for this approach within the broader tertiary environment.

Concerns about the rollout of broadband : a legal consideration of Australia’s NBN greenfield policy

This paper examines the Australian federal government’s proposal that developers take the primary role for deploying the National Broadband Network (‘NBN’) in greenfield estates. It identifies issues facing the NBN’s implementation and concerns raised by industry. A failure to address these concerns may lessen industry support as well as adversely impact on consumers as NBN implementation costs are passed onto them. The author identifies the need for NBN legislation to clearly establish what is a ‘greenfield estate’; how and when exemptions from implementation obligations will apply; and that NBN services must be treated the same as any other utility service.