Polyalanine repeat polymorphism in RUNX2 is associated with site-specific fracture in post-menopausal females

Runt related transcription factor 2 (RUNX2) is a key regulator of osteoblast differentiation. Several variations within RUNX2 have been found to be associated with significant changes in BMD, which is a major risk factor for fracture. In this study we report that an 18bp deletion within the polyalanine tract (17A>11A) of RUNX2 is significantly associated with fracture. Carriers of the 11A allele were found to be nearly twice as likely to have sustained fracture. Within the fracture category, there was a significant tendency of 11A carriers to present with fractures of bones of intramembranous origin compared to bones of endochondral origin (p=0.005). In a population of random subjects, the 11A allele was associated with decreased levels of serum collagen cross links (CTx, p=0.01), suggesting decreased bone turnover. The transactivation function of the 11A allele was quantitatively decreased. Interestingly, we found no effect of the 11A allele on BMD at multiple skeletal sites, although these were not the sites where a relationship with fracture was most evident. These findings suggest that the 11A allele is a biologically relevant polymorphism that influences serum CTx and confers enhanced fracture risk in a site-selective manner related to intramembranous bone ossification.

Probabilistic assessment of herbicide runoff from Japanese rice paddies: The effects of local meteorological conditions and site-specific water management

Uncertainty assessments of herbicide losses from rice paddies in Japan associated with local meteorological conditions and water management practices were performed using a pesticide fate and transport model, PCPF-1, under the Monte Carlo (MC) simulation scheme. First, MC simulations were conducted for five different cities with a prescribed water management scenario and a 10-year meteorological dataset of each city. The effectiveness of water management was observed regarding the reduction of pesticide runoff. However, a greater potential of pesticide runoff remained in Western Japan. Secondly, an extended analysis was attempted to evaluate the effects of local water management and meteorological conditions between the Chikugo River basin and the Sakura River basin using uncertainty inputs processed from observed water management data. The results showed that because of more severe rainfall events, significant pesticide runoff occurred in the Chikugo River basin even when appropriate irrigation practices were implemented. © Pesticide Science Society of Japan.

Comparative analysis of FimB and FimE recombinase activity

FimB and FimE are site-specific recombinases, part of the λ integrase family, and invert a 314 bp DNA switch that controls the expression of type 1 fimbriae in Escherichia coli. FimB and FimE differ in their activity towards the fim switch, with FimB catalysing inversion in both directions in comparison to the higher-frequency but unidirectional on-to-off recombination catalysed by FimE. Previous work has demonstrated that FimB, but not FimE, recombination is completely inhibited in vitro and in vivo by a regulator, PapB, expressed from a distinct fimbrial locus. The aim of this work was to investigate differences between FimB and FimE activity by exploiting the differential inhibition demonstrated by PapB. The research focused on genetic changes to the fim switch that alter recombinase binding and its structural context. FimB and FimE still recombined a switch in which the majority of fimS DNA was replaced with a larger region of non-fim DNA. This demonstrated a minimal requirement for FimB and FimE recombination of the Fim binding sites and associated inverted repeats. With the original leucine-responsive regulatory protein (Lrp) and integration host factor (IHF)-dependent structure removed, PapB was now able to inhibit both recombinases. The relative affinities of FimB and FimE were determined for the four ‘half sites’. This analysis, along with the effect of extensive swaps and duplications of the half sites on recombination frequency, demonstrated that FimB recruitment and therefore subsequent activity was dependent on a single half site and its context, whereas FimE recombination was less stringent, being able to interact initially with two half sites with equally high affinity. While increasing FimB recombination frequencies failed to overcome PapB repression, mutations made in recombinase binding sites resulted in inhibition of FimE recombination by PapB. Overall, the data support a model in which the recombinases differ in loading order and co-operative interactions. PapB exploits this difference and FimE becomes susceptible when its normal loading is restricted or changed.

Creating new narratives through shared time and space : the performer/audience connection in multi-site dance events

Site-specific performance provides choices in audience experience via degrees of scale, proximity, levels of immersion and viewing perspectives. Beyond these choices, multi-site promenade events also form a connected audience/performer relationship in which moving together in time and space can produce a shared narrative and aesthetic sensibility of collective, yet individuated and shifting meanings. This paper interrogates this notion through audience/performer experiences in two separate multi-site, dance-led events. here/there/then/now occurred in four intimate sites within the Brisbane Powerhouse, providing a theatricalised platform for audiences to create linked narratives through open-ended and fragmented intertextuality. Accented Body, based on the concept of “the body as site and in site” and notions of connectivity, provided a more expansive platform for a similar, but heightened, shared engagement. Audiences traversed 6 outdoor and 2 indoor Brisbane sites moving to varying levels of a large complex. Eleven, predominantly interactive, screens provided links to other sites as well as to distributed presences in Seoul and London. The differentiation in scale and travel time between sites deepened the immersive experiences of audiences who reported transformative engagements with both site and architecture, accompanied by a sense of extended and yet quickened time.

Risk-based approach to the management of on-site wastewater treatment facilities

Background On-site wastewater treatment system (OWTS) siting, design and management has traditionally been based on site specific conditions with little regard to the surrounding environment or the cumulative effect of other systems in the environment. The general approach has been to apply the same framework of standards and regulations to all sites equally, regardless of the sensitivity, or lack thereof, to the receiving environment. Consequently, this has led to the continuing poor performance and failure of on-site systems, resulting in environmental and public health consequences. As a result, there is increasing realisation that more scientifically robust evaluations in regard to site assessment and the underlying ground conditions are needed. Risk-based approaches to on-site system siting, design and management are considered the most appropriate means of improvement to the current standards and codes for on-site wastewater treatment systems. The Project Research in relation to this project was undertaken within the Gold Coast City Council region, the major focus being the semi-urban, rural residential and hinterland areas of the city that are not serviced by centralised treatment systems. The Gold Coast has over 15,000 on-site systems in use, with approximately 66% being common septic tank-subsurface dispersal systems. A recent study evaluating the performance of these systems within the Gold Coast area showed approximately 90% were not meeting the specified guidelines for effluent treatment and dispersal. The main focus of this research was to incorporate strong scientific knowledge into an integrated risk assessment process to allow suitable management practices to be set in place to mitigate the inherent risks. To achieve this, research was undertaken focusing on three main aspects involved with the performance and management of OWTS. Firstly, an investigation into the suitability of soil for providing appropriate effluent renovation was conducted. This involved detailed soil investigations, laboratory analysis and the use of multivariate statistical methods for analysing soil information. The outcomes of these investigations were developed into a framework for assessing soil suitability for effluent renovation. This formed the basis for the assessment of OWTS siting and design risks employed in the developed risk framework. Secondly, an assessment of the environmental and public health risks was performed specifically related the release of contaminants from OWTS. This involved detailed groundwater and surface water sampling and analysis to assess the current and potential risks of contamination throughout the Gold Coast region. Additionally, the assessment of public health risk incorporated the use of bacterial source tracking methods to identify the different sources of fecal contamination within monitored regions. Antibiotic resistance pattern analysis was utilised to determine the extent of human faecal contamination, with the outcomes utilised for providing a more indicative public health assessment. Finally, the outcomes of both the soil suitability assessment and ground and surface water monitoring was utilised for the development of the integrated risk framework. The research outcomes achieved through this project enabled the primary research aims and objects to be accomplished. This in turn would enable Gold Coast City Council to provide more appropriate assessment and management guidelines based on robust scientific knowledge which will ultimately ensure that the potential environmental and public health impacts resulting from on-site wastewater treatment is minimised. As part of the implementation of suitable management strategies, a critical point monitoring program (CPM) was formulated. This entailed the identification of the key critical parameters that contribute to the characterised risks at monitored locations within the study area. The CPM will allow more direct procedures to be implemented, targeting the specific hazards at sensitive areas throughout Gold Coast region.

Molecular basis for enhancement of the meiotic DMC1 recombinase by RAD51 associated protein 1 (RAD51AP1)

Homologous recombination is needed for meiotic chromosome segregation, genome maintenance, and tumor suppression. RAD51AP1 (RAD51 associated protein 1) has been shown to interact with and enhance the recombinase activity of RAD51. Accordingly, genetic ablation of RAD51AP1 leads to enhanced sensitivity to and also chromosome aberrations upon DNA damage, demonstrating a role for RAD51AP1 in mitotic homologous recombination. Here we show physical association of RAD51AP1 with the meiosis-specific recombinase DMC1 and a stimulatory effect of RAD51AP1 on the DMC1-mediated D-loop reaction. Mechanistic studies have revealed that RAD51AP1 enhances the ability of the DMC1 presynaptic filament to capture the duplex-DNA partner and to assemble the synaptic complex, in which the recombining DNA strands are homologously aligned. We also provide evidence that functional cooperation is dependent on complex formation between DMC1 and RAD51AP1 and that distinct epitopes in RAD51AP1 mediate interactions with RAD51 and DMC1. Finally, we show that RAD51AP1 is expressed in mouse testes, and that RAD51AP1 foci colocalize with a subset of DMC1 foci in spermatocytes. These results suggest that RAD51AP1 also serves an important role in meiotic homologous recombination.

Identification of salivary N-glycoproteins and measurement of glycosylation site occupancy by boronate glycoprotein enrichment and liquid chromatography/electrospray ionization tandem mass spectrometry

RATIONALE Diseases including cancer and congenital disorders of glycosylation have been associated with changes in the site-specific extent of protein glycosylation. Saliva can be non-invasively sampled and is rich in glycoproteins, giving it the potential to be a useful biofluid for the discovery and detection of disease biomarkers associated with changes in glycosylation. METHODS Saliva was collected from healthy individuals and glycoproteins were enriched using phenylboronic acid based glycoprotein enrichment resin. Proteins were deglycosylated with peptide-N-glycosidase F and digested with AspN or trypsin. Desalted peptides and deglycosylated peptides were separated by reversed-phase liquid chromatography and detected with on-line electrospray ionization quadrupole-time-of-flight mass spectrometry using a 5600 TripleTof instrument. Site-specific glycosylation occupancy was semi-quantitatively determined from the abundance of deglycosylated and nonglycosylated versions of each given peptide. RESULTS Glycoprotein enrichment identified 67 independent glycosylation sites from 24 unique proteins, a 3.9-fold increase in the number of glycosylation sites identified. Enrichment of glycoproteins rather than glycopeptides allowed detection of both deglycosylated and nonglycosylated versions of each peptide, and thereby robust measurement of site-specific occupancy at 21 asparagines. Healthy individuals showed limited biological variability in occupancy, with partially modified sites having characteristics consistent with inefficient glycosylation by oligosaccharyltransferase. Inclusion of negative controls without enzymatic deglycosylation controlled for spontaneous chemical deamidation, and identified asparagines previously incorrectly annotated as glycosylated. CONCLUSIONS We developed a sample preparation and mass spectrometry detection strategy for rapid and efficient measurement of site-specific glycosylation occupancy on diverse salivary glycoproteins suitable for biomarker discovery and detection of changes in glycosylation occupancy in human disease.

Evoking poetics of memory through performing site: Naik Naik, a Malaysian Australian collaboration in the world heritage setting of Melaka

Memory, time and metaphor are central triggers for artists in exploring and shaping their creative work. This paper examines the place of artists as ‘memory-keepers’, and ‘memory-makers’, in particular through engagement with the time-based art of site-specific performance. Naik Naik (Ascent) was a multi-site performance project in the historic setting of Melaka, Malaysia, and is partially recaptured through the presence and voices of its collaborating artists. Distilled from moments recalled, this paper seeks to uncover the poetics of memory to emerge from the project; one steeped in metaphor rather than narrative. It elicits some of the complex and interdependent layers of experience revealed by the artists in Naik Naik; cultural, ancestral, historical, personal, instinctual and embodied memories connected to sound, smell, touch, sensation and light, in a spatiotemporal context for which site is the catalyst. The liminal nature of memory at the heart of Naik Naik, provides a shared experience of past and present and future, performatively interwoven.

here/there/then/now

here/there/then/now was a practice-led research project that brought together 10 independent artists in dance, music, theatre and visual/media arts to create a site-specific program within the walls of the Brisbane Powerhouse. The purpose was to explore how to best conceive flexible performance platforms, theatricalise site-specific work and engage new audiences through forms of promenade experience that could provide open choices on how and where to view it. The sold out season of 6 performances, which took place 14-19 May 2002, presented three discrete performance installations set in intimate parts of the building, each with their own aesthetic and communicative intention, culminating in a fourth in-theatre installation, where memories of the first three coalesced and were reinterrogated. Each site thereby investigated meaning-making via the moving body and its critical relationship with space and objects, in a dramatic re-contextualisation of traditional solo dance forms, now re-articulated through interdisciplinary practices. The benefit of this approach was the creation of a layered and multimodal experience that could be both shared and subsequently critiqued by performers and audience alike.

Eco-retrofitting with building integrated living systems

Building integrated living systems (BILS), such as green roofs and living walls, could mitigate many of the challenges presented by climate change and biodiversity protection. However, few if any such systems have been constructed, and current tools for evaluating them are limited, especially under Australian subtropical conditions. BILS are difficult to assess, because living systems interact with complex, changing and site-specific social and environmental conditions. Our past research in design for eco-services has confirmed the need for better means of assessing the ecological values of BILS - let alone better models for assessing their thermal and hydrological performance. To address this problem, a research project is being developed jointly by researchers at the Central Queensland University (CQ University) and the Queensland University of Technology (QUT), along with industry collaborators. A mathematical model under development at CQ University will be applied and tested to determine its potential for predicting their complex, dynamic behaviour in different contexts. However, the paper focuses on the work at QUT. The QUT school of design is generating designs for living walls and roofs that provide a range of ecosystem goods and services, or ‘eco-services’, for a variety of micro-climates and functional contexts. The research at QUT aims to develop appropriate designs, virtual prototypes and quantitative methods for assessing the potential multiple benefits of BILS in subtropical climates. It is anticipated that the CQ University model for predicting thermal behaviour of living systems will provide a platform for the integration of ecological criteria and indicators. QUT will also explore means to predict and measure the value of eco-services provided by the systems, which is still largely uncharted territory. This research is ultimately intended to facilitate the eco-retrofitting of cities to increase natural capital and urban resource security - an essential component of sustainability. The talk will present the latest range of multifunctional, eco-productive living walls, roofs and urban space frames and their eco-services.

The moment of exchange : risk, vulnerability, responsibility

Any cycle of production and exchange – be it economic, cultural or aesthetic – involves an element of risk. It involves uncertainty, unpredictability, and a potential for new insight and innovation (the boom) as well as blockages, crises and breakdown (the bust). In performance, the risks are plentiful – economic, political, social, physical and psychological. The risks people are willing to take depend on their position in the exchange (performer, producer, venue manager or spectator), and their aesthetic preferences. This paper considers the often uncertain, confronting or ‘risky’ moment of exchange between performer, spectator and culture in Live Art practices. Encompassing body art, autobiographical art, site-specific art and other sorts of performative intervention in the public sphere, Live Art eschews the artifice of theatre, breaking down barriers between art and life, artist and spectator, to speak back to the public sphere, and challenge assumptions about bodies, identities, memories, relationships and histories. In the process, Live Art frequently privileges an uncertain, confrontational or ‘risky’ mode of exchange between performer, spectator and culture, as a way of challenging power structures. This paper examines the moment of exchange in terms of risk, vulnerability, responsibility and ethics. Why the romance with ‘risky’ behaviours and exchanges? Who is really taking a risk? What risk? With whose permission (or lack thereof)? What potential does a ‘risky’ exchange hold to destabilise aesthetic, social or political norms? Where lies the fine line between subversive intervention in the public sphere and sheer self-indulgence? What are the social and ethical implications of a moment of exchange that puts bodies, beliefs or social boundaries at ‘risk’? In this paper, these questions are addressed with reference to historical and contemporary practices under the broadly defined banner of Live Art, from the early work of Abrovamic and Burden, through to contemporary Australian practitioners like Fiona McGregor.

Can't see the water for the trees

The work was both conceived and constructed in-situ within Gnombup Swamp a seasonal water body at Bremer Bay, Western Australia. The work interacts with site-specific conditions including wind patterns and a datum of seasonal water levels marks. The work is the result of collaboration between soil scientist Paula Deegan and Ian Weir. The installation was documented with a series of 30 still digital photographs, later animated in Microsoft Powerpoint.