A 96-well plate fluorescence assay for assessment of cellular permeability and active efflux in Salmonella enterica serovar Typhimurium and Escherichia coli

Multiply antibiotic-resistant (MAR) mutants of Escherichia coli and Salmonella enterica are characterized by reduced susceptibility to several unrelated antibiotics, biocides and other xenobiotics. Porin loss and/or active efflux have been identified as a key mechanisms of MAR. A single rapid test was developed for MAR. The intracellular accumulation of the fluorescent probe Hoechst (H) 33342 (bisbenzimide) by MAR mutants and those with defined disruptions in efflux pump and porin genes was determined in 96-well plate format. The accumulation of H33342 was significantly (P < 0.0001) reduced in MAR mutants of S. enterica serovar Typhimurium (n = 4) and E. coli (n = 3) by 41 +/- 8% and 17.3 +/- 7.2%, respectively, compared with their parental strains, which was reversed by the transmembrane proton gradient-collapsing agent carbonyl cyanide-m-chlorophenyl hydrazone (CCCP) and the efflux pump inhibitor phenylalanine-arginine-beta-naphthylamide (PA beta N). The accumulation of H33342 was significantly reduced in mutants of Salmonella Typhimurium with defined disruptions in genes encoding the porins OmpC, OmpF, OmpX and OmpW, but increased in those with disruptions in efflux pump components TolC, AcrB and AcrF. Reduced accumulation of H33342 in three other MAR mutants of Salmonella Typhimurium correlated with the expression of porin and efflux pump proteins. The intracellular accumulation of H33342 provided a sensitive and specific test for MAR that is cheap and relatively rapid. Differential sensitivity to CCCP and PA beta N provided a further means to phenotypically identify MAR mutants and the role of active efflux in each strain.

Multiple active levels in Bi12TiO20 crystals

The existence of multiple active levels in a photorefractive Bi12TiO20 crystal is here investigated at 514.5nm wavelength. We carry out two-wave mixing experiments using symmetrically incident beams of equal intensities. A large amplitude fast phase modulation in one of the beams reduces the fringes visibility and improves the detection of the generated frequency modulated signals in both (R and S) output directions. Diffraction efficiencies of the phase (photorefractive) and the absorption (photochromic) gratings are quantitatively computed as functions of the grating period. Results show that the absorption grating has two distinct components: one associated to the photorefractive trap density modulation and another related to local light-induced effects between different levels. The photorefractive grating was also investigated at 633nm and 594nm (besides 514.5nm) and a significant quenching of the photorefractive effect was observed at these wavelengths.

Active control in flexible plates with piezoelectric actuators using linear matrix inequalities

The study of algorithms for active vibrations control in flexible structures became an area of enormous interest, mainly due to the countless demands of an optimal performance of mechanical systems as aircraft, aerospace and automotive structures. Smart structures, formed by a structure base, coupled with piezoelectric actuators and sensor are capable to guarantee the conditions demanded through the application of several types of controllers. The actuator/sensor materials are composed by piezoelectric ceramic (PZT - Lead Zirconate Titanate), commonly used as distributed actuators, and piezoelectric plastic films (PVDF-PolyVinyliDeno Floride), highly indicated for distributed sensors. The design process of such system encompasses three main phases: structural design; optimal placement of sensor/actuator (PVDF and PZT); and controller design. Consequently, for optimal design purposes, the structure, the sensor/actuator placement and the controller have to be considered simultaneously. This article addresses the optimal placement of actuators and sensors for design of controller for vibration attenuation in a flexible plate. Techniques involving linear matrix inequalities (LMI) to solve the Riccati's equation are used. The controller's gain is calculated using the linear quadratic regulator (LQR). The major advantage of LMI design is to enable specifications such as stability degree requirements, decay rate, input force limitation in the actuators and output peak bounder. It is also possible to assume that the model parameters involve uncertainties. LMI is a very useful tool for problems with constraints, where the parameters vary in a range of values. Once formulated in terms of LMI a problem can be solved efficiently by convex optimization algorithms.

When 'listeners can't talk' : comparing active listening in opening sequences of telephone and online counselling

While there is growing use of online counselling, little is known about its interactional organisation and how it compares to telephone counselling. This is despite past research suggesting that both counsellors and clients report the impact of the different modalities on the presentation and management of the counselling interaction. This paper compares the interactional affordances of telephone and online web counselling in opening sequences on Kids Help Line, a 24-hour Australian counselling service for children and young people up to the age of 25. We examine two ways that counsellors show active listening through response tokens and formulations. The analysis describes how counsellors’ use of minimal response tokens facilitate the clients’ problem presentation and are used in the management of turn taking and sequence organisation. For example, counsellors use the response token Mm hm to show that they understand that the client’s unit of talk to is not yet complete, and to affirm or invite the client to continue speaking. Formulations in phone and web counselling are another way that counsellors display active listening to re-present stretches of the clients’ preceding talk. In phone and web counselling, however, the respective modalities can complicate matters of turn transition and sequence organisation. By examining actual phone and online counselling sessions, this paper offers empirical demonstrations of the interactional affordances of phone and online counselling, and shows how the institutional practice of active listening is accomplished across different counselling modalities

ATR-FTIR measurement of biomass components in phosphonium ionic liquids

Qualitative and quantitative measurements of biomass components dissolved in the phosphonium ionic liquids (ILs), trihexyltetradecylphosphonium chloride ([P66614]Cl) and tributylmethylphosphonium methylsulphate ([P4441]MeSO 4), are obtained using attenuated total reflectance-FTIR. Absorption bands related to cellulose, hemicelluloses, and lignin dissolution monitored in situ in biomass-IL mixtures indicate lignin dissolution in both ILs and some holocellulose dissolution in the hydrophilic [P4441]MeSO 4. The kinetics of lignin dissolution reported here indicate that while dissolution in the hydrophobic IL [P66614]Cl appears to follow an accepted mechanism of acid catalyzed -aryl ether cleavage, dissolution in the hydrophilic IL [P4441]MeSO 4 does not appear to follow this mechanism and may not be followed by condensation reactions (initiated by reactive ketones). The measurement of lignin dissolution in phosphonium ILs based on absorbance at 1510 cm 1 has demonstrated utility. When coupled with the gravimetric Klason lignin method, ATR-FTIR study of reaction mixtures can lead to a better understanding of the delignification process. © 2012 Copyright Taylor and Francis Group, LLC.

Clustering composite SaaS components in cloud computing using a grouping genetic algorithm

Recently, Software as a Service (SaaS) in Cloud computing, has become more and more significant among software users and providers. To offer a SaaS with flexible functions at a low cost, SaaS providers have focused on the decomposition of the SaaS functionalities, or known as composite SaaS. This approach has introduced new challenges in SaaS resource management in data centres. One of the challenges is managing the resources allocated to the composite SaaS. Due to the dynamic environment of a Cloud data centre, resources that have been initially allocated to SaaS components may be overloaded or wasted. As such, reconfiguration for the components’ placement is triggered to maintain the performance of the composite SaaS. However, existing approaches often ignore the communication or dependencies between SaaS components in their implementation. In a composite SaaS, it is important to include these elements, as they will directly affect the performance of the SaaS. This paper will propose a Grouping Genetic Algorithm (GGA) for multiple composite SaaS application component clustering in Cloud computing that will address this gap. To the best of our knowledge, this is the first attempt to handle multiple composite SaaS reconfiguration placement in a dynamic Cloud environment. The experimental results demonstrate the feasibility and the scalability of the GGA.

The neglected dimension of community liveability : impact on social connectedness and active ageing in higher density accommodation

Purpose This thesis is about liveability, place and ageing in the high density urban landscape of Brisbane, Australia. As with other major developed cities around the globe, Brisbane has adopted policies to increase urban residential densities to meet the main liveability and sustainability aim of decreasing car dependence and therefore pollution, as well as to minimise the loss of greenfield areas and habitats to developers. This objective hinges on urban neighbourhoods/communities being liveable places, which residents do not have to leave for everyday living. Community/neighbourhood liveability is an essential ingredient in healthy ageing in place and has a substantial impact upon the safety, independence and well-being of older adults. It is generally accepted that ageing in place is optimal for both older people and the state. The optimality of ageing in place generally assumes that there is a particular quality to environments or standard of liveability in which people successfully age in place. The aim of this thesis was to examine if there are particular environmental qualities or aspects of liveability that test optimality and to better understand the key liveability factors that contribute to successful ageing in place. Method A strength of this thesis is that it draws on two separate studies to address the research question of what makes high density liveable for older people. In Chapter 3, the two methods are identified and differentiated as Method 1 (used in Paper 1) and Method 2 (used in Papers 2, 3, 4 and 5). Method 1 involved qualitative interviews with 24 inner city high density Brisbane residents. The major strength of this thesis is the innovative methodology outlined in the thesis as Method 2. Method 2 involved a case study approach employing qualitative and quantitative methods. Qualitative data was collected using semi-structured, in-depth interviews and time-use diaries completed by participants during the week of tracking. The quantitative data was gathered using Global Positioning Systems for tracking and Geographical Information Systems for mapping and analysis of participants’ activities. The combination of quantitative and qualitative analysis captured both participants’ subjective perceptions of their neighbourhoods and their patterns of movement. This enhanced understanding of how neighbourhoods and communities function and of the various liveability dimensions that contribute to active ageing and ageing in place for older people living in high density environments. Both studies’ participants were inner-city high density residents of Brisbane. The study based on Method 1 drew on a wider age demographic than the study based on Method 2. Findings The five papers presented in this thesis by publication indicate a complex inter-relationship of the factors that make a place liveable. The first three papers identify what is comparable and different between the physical and social factors of high density communities/neighbourhoods. The last two papers explore relationships between social engagement and broader community variables such as infrastructure and the physical built environments that are risk or protective factors relevant to community liveability, active ageing and ageing in place in high density. The research highlights the importance of creating and/or maintaining a barrier-free environment and liveable community for ageing adults. Together, the papers promote liveability, social engagement and active ageing in high density neighbourhoods by identifying factors that constitute liveability and strategies that foster active ageing and ageing in place, social connections and well-being. Recommendations There is a strong need to offer more support for active ageing and ageing in place. While the data analyses of this research provide insight into the lived experience of high density residents, further research is warranted. Further qualitative and quantitative research is needed to explore in more depth, the urban experience and opinions of older people living in urban environments. In particular, more empirical research and theory-building is needed in order to expand understanding of the particular environmental qualities that enable successful ageing in place in our cities and to guide efforts aimed at meeting this objective. The results suggest that encouraging the presence of more inner city retail outlets, particularly services that are utilised frequently in people’s daily lives such as supermarkets, medical services and pharmacies, would potentially help ensure residents fully engage in their local community. The connectivity of streets, footpaths and their role in facilitating the reaching of destinations are well understood as an important dimension of liveability. To encourage uptake of sustainable transport, the built environment must provide easy, accessible connections between buildings, walkways, cycle paths and public transport nodes. Wider streets, given that they take more time to cross than narrow streets, tend to .compromise safety - especially for older people. Similarly, the width of footpaths, the level of buffering, the presence of trees, lighting, seating and design of and distance between pedestrian crossings significantly affects the pedestrian experience for older people and impacts upon their choice of transportation. High density neighbourhoods also require greater levels of street fixtures and furniture for everyday life to make places more useable and comfortable for regular use. The importance of making the public realm useful and habitable for older people cannot be over-emphasised. Originality/value While older people are attracted to high density settings, there has been little empirical evidence linking liveability satisfaction with older people’s use of urban neighbourhoods. The current study examined the relationships between community/neighbourhood liveability, place and ageing to better understand the implications for those adults who age in place. The five papers presented in this thesis add to the understanding of what high density liveable age-friendly communities/ neighbourhoods are and what makes them so for older Australians. Neighbourhood liveability for older people is about being able to age in place and remain active. Issues of ageing in Australia and other areas of the developed world will become more critical in the coming decades. Creating livable communities for all ages calls for partnerships across all levels of government agencies and among different sectors within communities. The increasing percentage of older people in the community will have increasing political influence and it will be a foolish government who ignores the needs of an older society.

Mobility and active ageing in suburban environments : findings from in-depth interviews and person-based GPS tracking

Background. Governments face a significant challenge to ensure that community environments meet the mobility needs of an ageing population. Therefore, it is critical to investigate the effect of suburban environments on the choice of transportation and its relation to participation and active ageing. Objective. This research explores if and how suburban environments impact older people’s mobility and their use of different modes of transport. Methods. Data derived from GPS tracking, travel diaries, brief questionnaires, and semistructured interviews were gathered from thirteen people aged from 56 to 87 years, living in low-density suburban environments in Brisbane, Australia. Results. The suburban environment influenced the choice of transportation and out-of-home mobility. Both walkability and public transportation (access and usability) impact older people’s transportation choices. Impracticality of active and public transportation within suburban environments creates car dependency in older age. Conclusion. Suburban environments often create barriers to mobility, which impedes older people’s engagement in their wider community and ability to actively age in place. Further research is needed to develop approaches towards age-friendly suburban environments which will encourage older people to remain active and engaged in older age.

People with ID as active participants in nursing research : a literature review.

Aim: The movement from a medical model of disability to a more social model implies an imperative to include the views of people with ID in research regarding their care. Contemporary quality processes in improving care require consumer involvement at many levels and in doing so have shown better outcomes. A New Zealand research study is being undertaken utilizing focus groups with people with ID to understand their experiences during a psychiatric inpatient admission. The primary focus of this presentation will concern the literature review, undertaken as part of the study, of research in which people with ID have participated. Method: The literature review was conducted using a variety of electronic databases and search terms to identify studies with people with ID as active participants. Results: Only a few studies have been undertaken with people with ID as participants. While these studies demonstrate numerous benefits in including the voice of the person with ID this still remains absent from much of the research discourse. Conclusion: It is accepted and indeed advocated that people with ID have the same rights as others in regard to choosing whether or not they wish to participate in research. High response rates in the few identified studies indicated that people with ID are eager to be consulted. It is recommended that the unique needs of people with ID be taken into account in the research evidence base for future services.

Importance of sustainability-related cost components in highway infrastructure : perspective of stakeholders in Australia

Highway infrastructure development typically requires major capital input. Unless planned properly, such requirements can cause serious financial constraints for investors. The push for sustainability adds a new dimension to the complexity of evaluating highway projects. Finding environmentally and socially responsible solutions for highway construction will improve its potential for acceptance by the society and in many instances the infrastructure's life span. Even so, the prediction and determination of a project's long-term financial viability can be a precarious exercise. Existing studies in this area have not indicated details of how to identify and deal with costs incurred in pursuing sustainability measures in highway infrastructure. This paper provides insights into the major challenges of implementing sustainability in highway project development in terms of financial concerns and obligations. It discusses the results from recent research through a literature study and a questionnaire survey of key industry stakeholders involved in highway infrastructure development. The research identified critical cost components relating to sustainability measures based on perspectives of industry stakeholders. All stakeholders believe sustainability related costs are an integral part of the decision making. However, the importance rating of these costs is relative to each stakeholder's core business objectives. This will influence the way these cost components are dealt with during the evaluation of highway investment alternatives and financial implications. This research encourages positive thinking among the highway infrastructure practitioners about sustainability. It calls for the construction industry to maximise sustainability deliverables while ensuring financial viability over the life cycle of highway infrastructure projects.

The role of defects, or active states, in surface electrochemistry with particular reference to gold in neutral solution

Metastable, active, or nonequilibrium states due to the presence of abnormal structures and various types of defects are well known in metallurgy. The role of such states at gold surfaces in neutral aqueous media (an important electrode system in the microsensor area) was explored using cyclic voltammetry. It was demonstrated that, as postulated in earlier work from this laboratory, there is a close relationship between premonolayer oxidation, multilayer hydrous oxide reduction and electrocatalytic behaviour in the case of this and other metal electrode systems. Some of the most active, and therefore most important, entities at surfaces (e.g., metal adatoms) are not readily imageable or detectable by high resolution surface microscopy techniques. Cyclic voltammetry, however, provides significant, though not highly specific, information about such species. The main conclusion is that further practical and theoretical work on active states of metal surfaces is highly desirable as their behaviour is not simple and is of major importance in many electrocatalytic processes.

Synthesis of complementary RNA by RNA-dependent RNA polymerases in plant extracts is independent of an RNA primer

RNA-dependent RNA polymerase (RDR) activities were readily detected in extracts from cauliflower and broccoli florets, Arabidopsis thaliana (L.) Heynh callus tissue and broccoli nuclei. The synthesis of complementary RNA (cRNA) was independent of a RNA primer, whether or not the primer contained a 3′ terminal 2′-O-methyl group or was phosphorylated at the 5′ terminus. cRNA synthesis in plant extracts was not affected by loss-of-function mutations in the DICER-LIKE (DCL) proteins DCL2, DCL3, and DCL4, indicating that RDRs function independently of these DCL proteins. A loss-of-function mutation in RDR1, RDR2 or RDR6 did not significantly reduce the amount of cRNA synthesis. This indicates that these RDRs did not account for the bulk RDR activities in plant extracts, and suggest that either the individual RDRs each contribute a fraction of polymerase activity or another RDR(s) is predominant in the plant extract. © CSIRO 2008.

Environmental factors associated with active living in retirement village residents : findings from an exploratory qualitative enquiry

This exploratory enquiry employs qualitative methods to advance knowledge and understanding of physical environmental attributes related to active living among residents of Australian retirement villages. Six focus groups (n = 51 residents) were held and participants described how their current, and subsequently ideal, retirement village and neighborhood supported active lifestyles. Thematic analysis revealed three key environmental factors associated with active living: a positive social environment within the village; services and facilities provided in the village and wider neighborhood; and the presence of suitable pedestrian infrastructure. The unique discovery that environmental factors of both the retirement village and the surrounding neighborhood were associated with residents’ active living raises many questions for study. Findings informed the development of a survey instrument, and further understanding in this area has the potential to contribute to the design and siting practices of senior housing complexes within neighborhoods.