Characterisation and application of a bovine U6 promoter for expression of short hairpin RNAs

Background
The use of small interfering RNA (siRNA) molecules in animals to achieve double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful method of sequence-specific gene knockdown. As DNA-based expression of short hairpin RNA (shRNA) for RNAi may offer some advantages over chemical and in vitro synthesised siRNA, a number of vectors for expression of shRNA have been developed. These often feature polymerase III (pol. III) promoters of either mouse or human origin.
Results
To develop a shRNA expression vector specifically for bovine RNAi applications, we identified and characterised a novel bovine U6 small nuclear RNA (snRNA) promoter from bovine sequence data. This promoter is the putative bovine homologue of the human U6-8 snRNA promoter, and features a number of functional sequence elements that are characteristic of these types of pol. III promoters. A PCR based cloning strategy was used to incorporate this promoter sequence into plasmid vectors along with shRNA sequences for RNAi. The promoter was then used to express shRNAs, which resulted in the efficient knockdown of an exogenous reporter gene and an endogenous bovine gene.
Conclusion
We have mined data from the bovine genome sequencing project to identify a functional bovine U6 promoter and used the promoter sequence to construct a shRNA expression vector. The use of this native bovine promoter in shRNA expression is an important component of our future development of RNAi therapeutic and transgenic applications in bovine species.

Wastewater irrigation: the state of play

As demand for fresh water intensifies, wastewater is frequently being seen as a valuable resource. Furthermore, wise reuse of wastewater alleviates concerns attendant with its discharge to the environment. Globally, around 20 million ha of land are irrigated with wastewater, and this is likely to increase markedly during the next few decades as water stress intensifies. In 1995, around 2.3 billion people lived in water-stressed river basins and this could increase to 3.5 billion by 2025. We review the current status of wastewater irrigation by providing an overview of the extent of the practice throughout the world and through synthesizing the current understanding of factors influencing sustainable wastewater irrigation. A theme that emerges is that wastewater irrigation is not only more common in water-stressed regions such as the Near East, but the rationale for the practice also tends to differ between the developing and developed worlds. In developing nations, the prime drivers are livelihood dependence and food security, whereas environmental agendas appear to hold greater sway in the developed world. The following were identified as areas requiring greater understanding for the long-term sustainability of wastewater irrigation: (i) accumulation of bioavailable forms of heavy metals in soils, (ii) environmental fate of organics in wastewater-irrigated soils, (iii) influence of reuse schemes on catchment hydrology, including transport of salt loads, (iv) risk models for helminth infections (pertinent to developing nations), (v) microbiological contamination risks for aquifers and surface waters, (vi) transfer efficiencies of chemical contaminants from soil to plants, (vii) health effects of chronic exposure to chemical contaminants, and (viii) strategies for engaging the public.

Reforming Australian public sector accounting : an episode of institutional thinking

In recent years in Australia, accounting reforms have been developed which have resulted in the application of commercial systems of accounting to diverse public sector organisations. The reforms, which include the requirement to recognise infrastructure and heritage resources as assets in financial reports, endorse financial notions of accountability and performance that have been traditionally applied within private sector, profit-seeking organisations. Such notions are applied to a range of public sector organisations for the first time, even though the primary missions or objectives of many of these organisations are social, rather than financial in orientation. This critical, interpretative case study, set within the context of not-for-profit public museums, seeks to enhance an understanding of public sector accounting change based on these unique social organisations. The study examines three aspects of the reforms, namely, their development, their promotion and their defence. This examination is undertaken using the ideas contained in Mary Douglas’ (1986) How Institutions Think as the key theoretical construct. The supplementary perspectives of problematisation and epistemic communities are used to assist in applying the primary theoretical construct by explaining how, and by whom, these reforms were advocated and implemented in this specific instance. The study shows how the interpretation and application of the statements comprising the conceptual framework have shaped the development, promotion and defence of detailed standards developed for specific public sector organisations. In doing so, the study addresses two key research questions: (1) How were financial notions of accountability and performance of Australian public sector organisations constructed during the period 1976-2001 and articulated in the CF, once its development began, within this reform period? (2) How were these notions and other concepts of financial reporting outlined in the CF interpreted and applied in the (i) development; (ii) promotion; and (iii) defence of detailed accounting standards for not-for-profit public museums in Australia during the period under investigation? The study demonstrates that the concepts of financial reporting outlined in the conceptual framework were used by a relatively small group of technical experts located in influential positions in accounting regulation and in other fields to justify the application of accrual accounting within diverse public sector organisations. During the period examined, only certain questions were posed and certain issues considered and many problems associated with the implementation of the reforms were not considered. Accordingly, a key finding of the study is that each aspect of the reform period was guided and constrained by institutional thinking. In addition, the study shows how the framework's content can be used to permit equally well-argued, but conflicting, accounting policies to be adopted and defended for the same items, indicating the framework to be of only limited value as a technical tool. This leads to another key finding of the study, namely, that the framework is best understood as a political tool, serving a crucial role in enabling accrual accounting reforms to be developed, promoted and defended within the public sector. Thus, the study seeks to offer an enhanced understanding of the nature and determinants of accounting change, and accordingly, it broadens an understanding of the use of the conceptual framework, as an institution, in developing, promoting and defending changes to accounting practice.

Investigation of the intersection of constraints model of motion perception

The direction and speed of motion of a one-dimensional (1-D) stimulus, such as a grating, presented within a circular aperture is ambiguous. This ambiguity, referred to as the Aperture Problem (Fennema & Thompson, 1979) results from (i) the inability to detect motion parallel to grating orientation, and (ii) the occlusion of border information, such as the ‘ends’ of the grating, by the surface forming the aperture, Adelson and Movshon's (1982) intcrsection-of-constraints (IOC) model of motion perception describes a two-stage method of disambiguating the motion of 1-D moving stimuli (e.g., gratings) to produce unambiguous motion of two-dimensional (2-D) objects (e.g., plaid patterns) made up of several 1-D components. Specifically, in the IOC model ambiguous 1-D motions extracted by Stage 1 component-selective mechanisms are integrated by Stage 2 pattern-selective mechanisms to produce unambiguous 2-D motion signals. ‘Integration’ in the context of the IOC model involves determining the single motion vector (i.e., combination of direction and speed) which is consistent with the I-D components of a 2-D object. Since the IOC model assumes that 2-D objects undergo pure translation (i.e., without distortion, rotation, etc.), the motion vector consistent with all 1-D components describes the motion of the 2-D object itself. Adelson and Movshon (1982) propose that neural implementation of the computation underlying the IOC model is reflected in the perception of coherent 2-D plaid motion reported when two separately-moving ‘component’ gratings are superimposed. Using these plaid patterns the present thesis assesses the IOC model in terms of its ability to account for the perception of 2-D motion in a variety of circumstances. In the first series of experiments it is argued that the unambiguous motion perceived for a single grating presented within a rectangular aperture (i.e., the Barberpole illusion; Wallach, 1976) reflects application of the IOC computation to the moving 1-D grating and the stationary boundary of the aperture. While contrary to the assumption which underlies the IOC model (viz., that integration occurs between moving 1-D stimuli), evidence consistent with the involvement of the IOC computation in mediating the Barberpole illusion (in which there is only one moving stimulus) is obtained by measuring plaid coherence as a function of aperture shape. It is found that rectangular apertures which bias perceived component motions in directions consistent with plaid direction facilitate plaid coherence, while rectangular apertures which bias perceived component motions in directions inconsistent with plaid direction disrupt plaid coherence. In the second series of experiments, perceived directions of motion of type I symmetrical, type I asymmetrical, and type II plaids are measured with the aim of investigating the deviations in plaid directions reported by Ferrera and Wilson (1990) and Yo and Wilson (1992). Perceived directions of both asymmetrical and type II plaids are shown to deviate away from lOC-predicted directions and towards mean component direction. Furthermore, the magnitude of these deviations is being proportional to the difference between lOC-predicted plaid direction and mean component direction. On the basis of these directional deviations, modification to the IOC model is proposed. In the modified IOC model it is argued that plaid perception involves (i) the activity of Stage 2 pattern-selective mechanisms (and the Stage 1 component-selective mechanisms which input into these pattern-selective mechanisms) involved in implementing the IOC computation, and (ii) component-selective mechanisms which influence plaid perception directly, and ‘extraneously’ to the IOC computation. In the third series of experiments the validity of this modified IOC model, as well as the validity of alternative one-stage models of plaid perception are assessed in relation to perceived directions of plaid-induced MAEs as a function of both plaid direction and mean component direction. It is found that plaid-induced MAEs are shifted away from directions opposite to lOC-predicted plaid direction towards the direction opposite to mean component direction. This pattern of results is taken to be consistent with the modified IOC model which predicts the activity, and adaptation both of mechanisms signalling plaid direction (via implementation of the IOC computation), and ‘extraneous-type’ component-selective mechanisms signalling component directions. Alternative one-stage models which predict the adaptation of only mechanisms signalling plaid direction (the feature-tracking model), or the adaptation only of mechanisms signalling component directions (the distribution-of-activity model), cannot account for the directions of plaid-induced MAEs reported. The ability of the modified IOC model to account for the perceived directions of (i) gratings in rectangular apertures, (ii) various types of plaid in circular apertures, and (iii) directions of plaid-induced MAEs, is interpreted as supporting the proposition that human motion perception is based on a parallel and distributed process involving Stage 2 pattern-selective mechanisms (and the Stage 1 component-selective mechanisms which input into these mechanisms) taken to implement the IOC computation, and component-selective mechanisms taken to provide an 'extraneous' direct contribution to motion perception.

Status of tailings dumps : let D5s go working in the past?

It was decided in the De Beers v Ataqua Mining (Pty) Ltd that ''tailings dumps'' created by mining companies before the Mineral and Petroleum Resources Development Act, 28 of 2002 ("the MPRDA") came into operation are not governed by its provisions because such dumps are not "residue stockpiles" or "residue deposits" for purposes of the MPRDA. Ownership of tailings dumps is determined by the common law principles of accession. Ownership of a movable dump has to be transferred by one of the recognised forms of delivery of movables. Processing of these dumps will, however, still be subject to compliance with South African environmental, health and safety laws in general. It is submitted that mine dumps or tailings dumps created upon the exercise of "old order mining rights" before the commencement of the MPRDA and even after commencement of the MPRDA until eventual termination of the "old order mining rights" are not subject to the extensive, mining, environmental, empowerment provisions of the MPRDA. Termination of "old order mining rights" takes place upon: (i) refusal of an application for conversion of a mining right during (or even after) the period of transition, (ii) conversion into and registration of new order mining rights during (or even after) the period of transition or (iii) termination of unconverted "old order mining rights" on 30 April 2009. To the extent that this decision has made it possible to embark on a shorter and less cumbersome route in the reprocessing and eventual disappearance of most tailings dumps, it is to be welcomed from an economical, environmental, job creation and aesthetic perspective. Proposed amendments to the MPRDA to undo the impact of the De Beers decision should be carefully considered against these mentioned benefits and a possible finding that it may amount to an expropriation without compensation.

Looking to Sweden in order to reconstruct Australia

The most prominent interest in the 'Swedish model' in Australian political history came after a high-level union delegation visit in 1986 produced a major report titled Australia Reconstructed, which had the nominal endorsement of the Labor Government then in office. However, at this very time, the characteristics of the Swedish policy approach which were most admired by the visiting Australian unionists were undergoing important changes, to which they paid little attention but which critics from the Right strongly emphasized in response.

The Australian labour movement interest in Sweden in the 1980s had some distinctive features. It focused on manufacturing industry and skills training policy, reflecting priorities of the main participating unionists. It was also particularly concerned with industrial democracy and work design, as a result of important earlier links between Swedish, Norwegian and Australian industrial relations scholars and practitioners which were forged from the late 1960s. Increased interest in Sweden partly arose from the search for a new political vision by particular elements of the Australian Left following their disillusionment with the Soviet Union after the 1968 Prague Spring.

This article presents the results of interviews with participants and extensive archival research to provide new information and perspective on Scandinavian influences on the Australian labour movement; and the political background of the main people involved.

It also analyzes how discussion of the 'Swedish model' receded in Australia following the economic setbacks of the early 1990s amid a perception that 'the model' had collapsed, but how policy interest in the continuing evident achievements of Sweden and the other Nordic nations has gradually re-emerged in Australia since then, though in a somewhat different way to before.

Depression as a predictor of work resumption following myocardial infarction (MI): a review of recent research evidence

Background Depression often coexists with myocardial infarction (MI) and has been found to impede recovery through reduced functioning in key areas of life such as work. In an era of improved survival rates and extended working lives, we review whether depression remains a predictor of poorer work outcomes following MI by systematically reviewing literature from the past 15 years.

Methods Articles were identified using medical, health, occupational and social science databases, including PubMed, OVID, Medline, Proquest, CINAHL plus, CCOHS, SCOPUS, Web of Knowledge, and the following pre-determined criteria were applied: (i) collection of depression measures (as distinct from 'psychological distress') and work status at baseline, (ii) examination and statistical analysis of predictors of work outcomes, (iii) inclusion of cohorts with patients exhibiting symptoms consistent with Acute Coronary Syndrome (ACS), (iv) follow-up of work-specific and depression specific outcomes at minimum 6 months, (v) published in English over the past 15 years. Results from included articles were then evaluated for quality and analysed by comparing effect size.

Results Of the 12 articles meeting criteria, depression significantly predicted reduced likelihood of return to work (RTW) in the majority of studies (n = 7). Further, there was a trend suggesting that increased depression severity was associated with poorer RTW outcomes 6 to 12 months after a cardiac event. Other common significant predictors of RTW were age and patient perceptions of their illness and work performance.

Conclusion Depression is a predictor of work resumption post-MI. As work is a major component of Quality of Life (QOL), this finding has clinical, social, public health and economic implications in the modern era. Targeted depression interventions could facilitate RTW post-MI.

Improving load forecasting accuracy through combination of best forecasts

Neural network (NN) models have been widely used in the literature for short-term load forecasting. Their popularity is mainly due to their excellent learning and approximation capability. However, their forecasting performance significantly depends on several factors including initializing parameters, training algorithm, and NN structure. To minimize negative effects of these factors, this paper proposes a practically simple, yet effective and an efficient method to combine forecasts generated by NN models. The proposed method includes three main phases: (i) training NNs with different structures, (ii) selecting best NN models based on their forecasting performance for a validation set, and (iii) combination of forecasts for selected best NNs. Forecast combination is performed through calculating the mean of forecasts generated by best NN models. The performance of the proposed method is examined using real world data set. Comparative studies demonstrate that the accuracy of combined forecasts is significantly superior to those obtained from individual NN models.

Social work and faith-based agencies in Sweden and Australia

The roles which faith-based agencies play in social work provision vary between countries. This article provides an overview of social work provision by the Church of Sweden in Sweden and the Catholic Church in Australia and explores how different relationships between faith-based organizations and professional social work practice have emerged in different countries. The article concludes with questions about the role of faith-based agencies which readers can reflect upon in their own contexts.

Six million in Melbourne or a network of sustainable midi-cities? A thought experiment

By 2050, it is projected that Melbourne will have a population of between 5.6 and 6.4 million (DPCD, 2012), an increase of nearly 50% above its current level. Despite Melbourne's status as the world's most liveable city, a recent survey found that Australians in general found smaller cities are better places to live and bring up families (Perkins, 2013). The Grattan Institute's report entitled "The Cities We Need" was "an invitation to a conversation" about our future cities (Kelly, 2010:5). One idea not canvassed in the report was that of decentralization to accommodate Melbourne's projected growth. In its discussion paper, "Let's Talk about the Future", the Victorian State Government proposes that Melbourne become a 'polycentric city' linked to its regional cities (DPCD, 2012). While growth in the present regional cities is acknowledged, the possibility that these and other new regional cities could absorb the future population projected for Melbourne is not considered, nor that these regional cities could be transformed into 'sustainable cities'. This paper explores the idea that a network of smaller 'midi-cities, based on the sustainable city concept of Sweden, might provide a better alternative to concentrated growth in one city. Fifteen new cities of 150,000 would be required to absorb the projected extra 2.3 million Victorian residents. The paper analyses the energy, food, water and land requirements of a typical sustainable city. The new cities would require approximately 12% of the State's land area for food and energy supply, as well as the built environment.

Implementation of pressure ulcer prevention best practice recommendations in acute care: an observational study

Pressure ulcers are a common but preventable problem in hospitals. Implementation of best practice guideline recommendations can prevent ulcers from occurring. This 9-year cohort study reports prevalence data from point prevalence surveys during the observation period, and three practice metrics to assess implementation of best practice guideline recommendations: (i) nurse compliance with use of a validated pressure ulcer risk assessment and intervention checklist; (ii) accuracy of risk assessment scoring in usual-care nurses and experienced injury prevention nurses; and (iii) use of pressure ulcer prevention strategies. The prevalence of hospital-acquired pressure ulcers decreased following implementation of an evidence-based prevention programme from 12·6% (2 years preprogramme implementation) to 2·6% (6 years postprogramme implementation) (P < 0·001). Audits between 2003 and 2011 of 4368 patient medical records identified compliance with pressure ulcer prevention documentation according to best practice guidelines was high (>84%). A sample of 270 patients formed the sample for the study of risk assessment scoring accuracy and use of prevention strategies. It was found usual-care nurses under-estimated patients' risk of pressure ulcer development and under-utilised prevention strategies compared with experienced injury prevention nurses. Despite a significant reduction in prevalence of hospital-acquired pressure ulcers and high documentation compliance, use of prevention strategies could further be improved to achieve better patient outcomes. Barriers to the use of prevention strategies by nurses in the acute hospital setting require further examination. This study provides important insights into the knowledge translation of pressure ulcer prevention best practice guideline recommendations at The Northern Hospital.

Roles of additives in the trihexyl(tetradecyl) phosphonium chloride ionic liquid electrolyte for primary mg-air cells

As reported previously, water saturated trihexyl(tetradecyl)phosphonium chloride ([P6,6,6,14][Cl]) ionic liquid (IL) is a promising electrolyte for magnesium-air batteries. The added water plays an important role in enabling high rate and high efficiency Mg dissolution while stabilizing the Mg interphase. In this work, the role of the water was investigated by replacement with other additives such as toluene and tetrahydrofuran to specifically target the assumed roles of water, namely: (i) enhancement of transport properties; (ii) complexation and stabilization of the Mg anode; (iii) provision of active protons for the cathodic reaction. Discharge tests show that ethylene glycol supports comparable performance to that provided by water. Examination of the viscosity and conductivity of different [P6,6,6,14][Cl]/additive mixtures indicates that a simple consideration of solution characteristics cannot explain the observed trends. Rather, other factors, such as the presence of active protons and/or oxygen-donor groups, are also key features for the development of IL electrolytes for practical magnesium-air cells. Finally, the presence of ethylene glycol in the electrolyte results in a complex gel on the Mg interface, similar to that found in the presence of water. This may also play a role in enabling stable discharge of the Mg anode. © 2014 The Electrochemical Society.

Long-distance endozoochorous dispersal of submerged macrophyte seeds by migratory waterbirds in northern Europe - a critical review of possibilities and limitations

We review whether migratory Anatidae, i.e., swans, geese and ducks, could be acting as vectors for dispersal of Zostera, Ruppia and Potamogeton propagules by endozoochory (carrying seeds in their guts). We list six prerequisites that must all be fulfilled, if successful dispersal should occur. Several Anatidae species feed on these macrophytes, and undertake rapid long-distance movements, making dispersal possible. We identify four problems, which in combination leads us to conclude that long-distance dispersal events are likely to be rare. (i) Most long-distance movements are out of phase with the reproductive efforts of the plants, and if birds arrive at sites when plants still bear seeds, they are likely to depart well after seed stocks have been depleted. (ii) Seed transport by birds will usually be uni-directional, from north to south on autumn migrations. (iii) Most of the gut contents of migratory birds are likely to have been discarded within 300 km of departure. (iv) In many cases, birds will arrive in habitats seriously different from those they departed, i.e., any seeds carried along will have low chances of surviving in their new site. We suggest that northbound dispersal by endozoochory can only occur during spring if waterbirds feed on seeds that have not been depleted and remained frozen down or buried in sediments, or during moult- or post-moult migrations. Moult migration takes place in summer in phase with the reproductive efforts of the plants. Also epizoochorous dispersal (external attachment) is subject to restrictions i, ii and iv.