943 resultados para Green Fluorescent Protein


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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.

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Precise protein quantification is essential in clinical dietetics, particularly in the management of renal, burn and malnourished patients. The EP-10 was developed to expedite the estimation of dietary protein for nutritional assessment and recommendation. The main objective of this study was to compare the validity and efficacy of the EP-10 with the American Dietetic Association’s “Exchange List for Meal Planning” (ADA-7g) in quantifying dietary protein intake, against computerised nutrient analysis (CNA). Protein intake of 197 food records kept by healthy adult subjects in Singapore was determined thrice using three different methods – (1) EP-10, (2) ADA-7g and (3) CNA using SERVE program (Version 4.0). Assessments using the EP-10 and ADA-7g were performed by two assessors in a blind crossover manner while a third assessor performed the CNA. All assessors were blind to each other’s results. Time taken to assess a subsample (n=165) using the EP-10 and ADA-7g was also recorded. Mean difference in protein intake quantification when compared to the CNA was statistically non-significant for the EP-10 (1.4 ± 16.3 g, P = .239) and statistically significant for the ADA-7g (-2.2 ± 15.6 g, P = .046). Both the EP-10 and ADA-7g had clinically acceptable agreement with the CNA as determined via Bland-Altman plots, although it was found that EP-10 had a tendency to overestimate with protein intakes above 150 g. The EP-10 required significantly less time for protein intake quantification than the ADA-7g (mean time of 65 ± 36 seconds vs. 111 ± 40 seconds, P < .001). The EP-10 and ADA-7g are valid clinical tools for protein intake quantification in an Asian context, with EP-10 being more time efficient. However, a dietician’s discretion is needed when the EP-10 is used on protein intakes above 150g.

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Buildings are one of the most significant infrastructures in modern societies. The construction and operation of modern buildings consume a considerable amount of energy and materials, therefore contribute significantly to the climate change process. In order to reduce the environmental impact of buildings, various green building rating tools have been developed. In this paper, energy uses of the building sector in Australia and over the world are first reviewed. This is then followed by discussions on the development and scopes of various green building rating tools, with a particular focus on the Green Star rating scheme developed in Australia. It is shown that Green Star has significant implications on almost every aspect of the design of HVAC systems, including the selection of air handling and distribution systems, fluid handling systems, refrigeration systems, heat rejection systems and building control systems.

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This paper investigates energy saving potential of commercial building by living wall and green façade system using Envelope Thermal Transfer Value (ETTV) equation in Sub-tropical climate of Australia. Energy saving of four commercial buildings was quantified by applying living wall and green façade system to the west facing wall. A field experimental facility, from which temperature data of living wall system was collected, was used to quantify wall temperatures and heat gain under controlled conditions. The experimental parameters were accumulated with extensive data of existing commercial building to quantify energy saving. Based on temperature data of living wall system comprised of Australian native plants, equivalent temperature of living wall system has been computed. Then, shading coefficient of plants in green façade system has been included in mathematical equation and in graphical analysis. To minimize the air-conditioned load of commercial building, therefore to minimize the heat gain of commercial building, an analysis of building heat gain reduction by living wall and green façade system has been performed. Overall, cooling energy performance of commercial building before and after living wall and green façade system application has been examined. The quantified energy saving showed that only living wall system on opaque part of west facing wall can save 8-13 % of cooling energy consumption where as only green façade system on opaque part of west facing wall can save 9.5-18% cooling energy consumption of commercial building. Again, green façade system on fenestration system on west facing wall can save 28-35 % of cooling energy consumption where as combination of both living wall on opaque part of west facing wall and green façade on fenestration system on west facing wall can save 35-40% cooling energy consumption of commercial building in sub-tropical climate of Australia.

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This paper investigates cooling energy performance of commercial building before and after green roof and living wall application based on integrated building heat gain model developed from Overall Thermal Transfer Value (OTTV) of building wall and steady state heat transfer process of roof in sub-tropical climate. Using the modelled equation and eQUEST energy simulation tool, commercial building envelope parameters and relevant heat gain parameters have been accumulated to analyse the heat gain and cooling energy consumption of commercial building. Real life commercial building envelope and air-conditioned load data for the sub-tropical climate zone have been collected and compared with the modelled analysis. Relevant temperature data required for living wall and green roof analysis have been collected from experimental setup comprised of both green roof and west facing living wall. Then, Commercial building heat flux and cooling energy performance before and after green roof and living wall application have been scrutinized.

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This report discusses findings of a case study into "Green Buildings" undertaken as a part of the retrospective analysis component of Sustainable Built Environment National Research Centre (SBEnrc) Project 2.7 Leveraging R&D investment for the Australian Built Environment. The Western Australian Government (WAG) has taken a leadership role for a number of decades in developing more environmentally responsive buildings. In the past decade, considerable initiatives have been introduced to contribute to: (i) greening the stock of government buildings; and (ii) providing leadership in the development of other non-residential buildings developed commercially. This role has been informed by global, national and internal initiatives and research in this area. This case study investigates: (i) the nature of this leadership; and (ii) the role of R&D policy development; and (iii) the dissemination and impact of outcomes in the broader industry. This case study should be read in conjunction with Part 1 of this suite of reports.

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Purpose With an increasingly ageing population and widespread acceptance of the need for sustainable development in Australia, the demand for green retirement villages is increasing. This paper aims to identify the critical issues to be considered by developers and practitioners when embarking on their first green residential retirement project in Australia. Design/methodology/approach In view of the lack of adequate historical data for quantitative analysis, a case study approach is employed to examine the successful delivery of green retirement villages. Face-to-face interviews and document analysis were conducted for data collection. Findings The findings of the study indicate that one of the major obstacles to the provision of affordable green retirement villages is the higher initial costs involved. However, positive aspects were identified, the most significant of which relate to: the innovative design of site and floor plans; adoption of thermally efficient building materials; orientation of windows; installation of water harvesting and recycling systems, water conservation fittings and appliances; and waste management during the construction stage. With the adoption of these measures, it is believed that sustainable retirement development can be achieved without significant additional capital costs. Practical implications The research findings serve as a guide for developers in decision making throughout the project life-cycle when introducing green features into the provision of affordable retirement accommodation. Originality/value This paper provides insights into the means by which affordable green residential retirement projects for aged people can be successfully completed.

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There has been substantial interest within the Australian sugar industry in product diversification as a means to reduce its exposure to fluctuating raw sugar prices and in order to increase its commercial viability. In particular, the industry is looking at fibrous residues from sugarcane harvesting (trash) and from sugarcane milling (bagasse) for cogeneration and the production of biocommodities, as these are complementary to the core process of sugar production. A means of producing surplus residue (biomass) is to process whole sugarcane crop. In this paper, the composition of different juices derived from different harvesting methods, viz. burnt cane with all trash extracted (BE), green cane with half of the trash extracted (GE), and green cane (whole sugarcane crop) with trash unextracted (GU), were investigated and the results and comparison presented. The determination of electrical conductivity, inorganic composition, and organic acids indicate that both GU and GE cane juice contain a higher proportion of soluble inorganic ions and ionisable organic acids, compared to BE cane juice. It is important to note that there are considerably higher levels of Na ions and citric acid, but relatively low P levels in the GU samples. A higher level of reducing sugars was analysed in the GU samples than the BE samples due to the higher proportion of impurities found naturally in sugarcane tops and leaves. The purity of the first expressed juice (FEJ) of GU cane was on average higher than that of FEJ of BE cane. Results also show that GU juices appear to contain higher levels of proteins and polysaccharides, with no significant difference in starch levels.

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Background: The term ‘green health promotion’ is given to health promotion underpinned by the principles of ecological health and sustainability. Green health promotion is supported philosophically by global health promotion documents such as the Ottawa Charter for Health Promotion (1986) and the ecological public health movement. Green health promotion in schools aims to practice the principles of ecological health and sustainability. Methods: An extended literature review revealed a paucity of literature about green health promotion in schools across disciplines. Literature about nurses and health promotion in schools is generally found in nursing publications. Literature about ecological sustainability in schools is mostly found in teaching publications. Results: This paper explores the nexus between nursing and health promotion, and teachers and ecological sustainability. Collaborative partnerships between health and education do not capitalise on programs such as Health Promoting Schools and the School Based Youth Health Nurse Program in Queensland, Australia. The authors consider how collaborative partnerships between health and education in schools can work towards green health promotion. Conclusion: Nursing’s approach to health promotion and education’s approach to ecological sustainability need to be aligned to enhance green health promotion in schools and promote a new generation of ‘tree huggers and hippies’.

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Ubiquitylation is a necessary step in the endocytosis and lysosomal trafficking of many plasma membrane proteins and can also influence protein trafficking in the biosynthetic pathway. Although a molecular understanding of ubiquitylation in these processes is beginning to emerge, very little is known about the role deubiquitylation may play. Fat Facets in mouse (FAM) is substrate-specific deubiquitylating enzyme highly expressed in epithelia where it interacts with its substrate, β-catenin. Here we show, in the polarized intestinal epithelial cell line T84, FAM localized to multiple points of protein trafficking. FAM interacted with β-catenin and E-cadherin in T84 cells but only in subconfluent cultures. FAM extensively colocalized with β-catenin in cytoplasmic puncta but not at sites of cell-cell contact as well as immunoprecipitating with β-catenin and E-cadherin from a higher molecular weight complex (~500 kDa). At confluence FAM neither colocalized with, nor immunoprecipitated, β-catenin or E-cadherin, which were predominantly in a larger molecular weight complex (~2 MDa) at the cell surface. Overexpression of FAM in MCF-7 epithelial cells resulted in increased β-catenin levels, which localized to the plasma membrane. Expression of E-cadherin in L-cell fibroblasts resulted in the relocalization of FAM from the Golgi to cytoplasmic puncta. These data strongly suggest that FAM associates with E-cadherin and β-catenin during trafficking to the plasma membrane.

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Only a few years ago there were only a handful of buildings in Australia, mainly leased by or from the Commonwealth Government to which a green lease might have application. Now with the passing of the Building Energy Efficiency Disclosure Act 2010 (Cth) all commercial office premises in excess of 2000 square metres have 12 months from 1 November 2010 to obtain a Building Energy Efficiency Certificate as part of Stage 1 of the Federal Government’s National Framework for Energy Efficiency This significant change has focused attention on changes required to the conditions of leases where the building has a NABERS rating. This article considers material from the United Kingdom, the United States and Canada where there are similar policy changes in play and makes suggestions as to how certain clauses of a standard lease of a commercial office block may be altered to meet this new regime.

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Public road authorities have a key responsibility in driving initiatives for reducing greenhouse gas (GHG) emissions in the road construction project lifecycle. A coherent and efficient chain of procurement processes and methods is needed to convert green policies into tangible actions that capture the potential for GHG reduction. Yet, many infrastructure clients lack developed methodologies regarding green procurement practices. Designing more efficient solutions for green procurement requires an evaluation of the current initiatives and stages of development. A mapping of the current GHG reduction initiatives in Australian public road procurement is presented in this paper. The study includes the five largest Australian state road authorities, which cover 94% of the total 817,089 km of Australian main roads (not local) and account for 96% of the total A$13 billion annual major road construction and maintenance expenditure. The state road authorities’ green procurement processes and tools are evaluated based on interviews and a review of documents. Altogether 12 people, comprising 1-3 people of each organisation, participated in the interviews and provided documents. An evaluation matrix was developed for mapping the findings across the lifecycle of road construction project delivery. The results show how Australian state road authorities drive decisions with an impact on GHG emissions on the strategic planning phase, project development phase, and project implementation phase. The road authorities demonstrate varying levels of advancement in their green procurement methodologies. Six major gaps in the current green procurement processes are identified and, respectively, six recommendations for future research and development are suggested. The greatest gaps remain in the project development phase, which has a critical role in fixing the project (GHG reduction) goals, identifying risks and opportunities, and selecting the contractor to deliver the project. Specifically, the role of mass-haul optimisation as a part of GHG minimisation was reviewed, and mass-haul management was found to be an underutilised element with GHG reduction potential.

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Green building is building that the focus is to maximize the energy efficiency and resources used. While, retrofitting is the process of renovate or refurnish the existing building. Therefore by retrofit existing buildings that comply with green building requirement, it improves the environmental attributes of the buildings. In Malaysia, existing buildings and its communities contribute over 40% of green house gases to the environment. This paper describes a study that explores the potential to retrofit existing campus buildings that response to sustainable green building standard. A validation survey was carried out and the data collected was analysed using SPSS in order to confirm the significance of retrofitting Universiti Teknologi Malaysia (UTM) buildings toward green building initiative. The results show that all the twenty eight identified green elements recorded average index of higher than 3.5 which means that there is significant needs to retrofit the existing buildings to green buildings. This study concludes that it is urgently need for the campus to response to green building requirements in order to achieve higher energy effeciency and this can be done through effective etrofitting of existing buildings.

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Reciprocal interactions between Src family kinases (SFKs) and focal adhesion kinase (FAK) are critical during changes in cell attachment. Recently it has been recognized that another SFK substrate, CUB-domain-containing protein 1 (CDCP1), is differentially phosphorylated during these events. However, the molecular processes underlying SFK-mediated phosphorylation of CDCP1 are poorly understood. Here we identify a novel mechanism in which FAK tyrosine 861 and CDCP1-Tyr-734 compete as SFK substrates and demonstrate cellular settings in which SFKs switch between these sites. Our results show that stable CDCP1 expression induces robust SFK-mediated phosphorylation of CDCP1-Tyr-734 with concomitant loss of p-FAK-Tyr-861 in adherent HeLa cells. SFK substrate switching in these cells is dependent on the level of expression of CDCP1 and is also dependent on CDCP1-Tyr-734 but is independent of CDCP1-Tyr-743 and -Tyr-762. In HeLa CDCP1 cells, engagement of SFKs with CDCP1 is accompanied by an increase in phosphorylation of Src-Tyr-416 and a change in cell morphology to a fibroblastic appearance dependent on CDCP1-Tyr-734. SFK switching between FAK-Tyr-861 and CDCP1-Tyr-734 also occurs during changes in adhesion of colorectal cancer cell lines endogenously expressing these two proteins. Consistently, increased p-FAK-Tyr-861 levels and a more epithelial morphology are seen in colon cancer SW480 cells silenced for CDCP1. Unlike protein kinase Cδ, FAK does not appear to form a trimeric complex with Src and CDCP1. These data demonstrate novel aspects of the dynamics of SFK-mediated cell signaling that may be relevant during cancer progression.

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This paper reports on a current case study of green building initiatives implemented by the Western Australian government in the past decade. The intent is to provide a qualitative understanding of past R&D investments in the Australian built environment. The case method was selected to illustrate three sector-based investments, one of which is reported on here. The conceptual framework underpinning interview design and data analysis uses dynamic capability, absorptive capacity and open innovation theories to better understand the organisational environment in which these initiatives were implemented. Data has been thematically coded to criteria identified from the literature to illustrate organisational characteristics which may have contributed to dissemination and impact. The results will be combined with two further case studies (construction safety and digital modelling), to inform this research. This industry supported project will conclude by developing policy guidelines for future R&D investment in the built environment.