Cloning, expression, characterisation and mutational analysis of l-aspartate oxidase from Pseudomonas putida

L-Amino acid oxidases (LAAOs) are useful catalysts for the deracemisation of racemic amino acid sub-strates when combined with abiotic reductants. The gene nadB encoding the L-aspartate amino acid oxidase from Pseudomonas putida (PpLASPO) has been cloned and expressed in E. coli. The purified PpLASPO enzyme displayed a K M for l-aspartic acid of 2.26 mM and a k cat = 10.6 s −1 , with lower activity also displayed towards L-asparagine, for which pronounced substrate inhibition was also observed. The pH optimum of the enzyme was recorded at pH 7.4. The enzyme was stable for 60 min at up to 40 • C, but rapid losses in activity were observed at 50 • C. A mutational analysis of the enzyme, based on its sequence homology with the LASPO from E. coli of known structure, appeared to confirm roles in substrate binding or catalysis for residues His244, His351, Arg386 and Arg290 and also for Thr259 and Gln242. The high activity of the enzyme, and its promiscuous acceptance of both L-asparagine and L-glutamate as substrates, if with low activity, suggests that PpLASPO may provide a good model enzyme for evolution studies towards AAOs of altered or improved properties in the future.

Experimental and numerical analysis of the relationship between indoor and outdoor airborne particles in an operating room

This work investigated the impact of the HVAC filtration system and indoor particle sources on the relationship between indoor and outdoor airborne particle size and concentrations in an operating room. Filters with efficiency between 65% and 99.97% were used in the investigation and indoor and outdoor particle size and concentrations were measured. A balance mass model was used for the simulation of the impact of the surgical team, deposition rate, HVAC exhaust and air change rates on indoor particle concentration. The experimental results showed that high efficiency filters would not be expected to decrease the risk associated with indoor particles larger than approximately 1 µm in size because normal filters are relatively efficient for these large particles. A good fraction of outdoor particles were removed by deposition on the HVAC system surfaces and this deposition increased with particle size. For particles of 0.3-0.5 µm in diameter, particle reduction was about 23%, while for particles >10 µm the loss was about 78%. The modelling results showed that depending on the type of filter used, the surgical team generated between 93-99% of total particles, while the outdoor air contributed only 1-6%.

Molecular and cellular analysis of basement membrane invasion by human breast cancer cells in Matrigel-based in vitro assays

In vitro analyses of basement membrane invasiveness employing Matrigel (a murine tumor extract rich in basement membrane components) have been performed on human breast cancer model systems. Constitutive invasiveness of different human breast cancer (HBC) cell lines has been examined as well as regulation by steroid hormones, growth factors, and oncogenes. Carcinoma cells exhibiting a mesenchymal-like phenotype (vimentin expression, lack of cell border associated uvomorulin) show dramatically increased motility, invasiveness, and metastatic potential in nude mice. These findings support the hypothesis that epithelial to mesenchymal transition (EMT)-like events may be instrumental in the metastatic progression of human breast cancer. The MCF-7 subline MCF-7ADR appears to have undergone such a transition. The importance of such a transition may be reflected in the emergence of vimentin expression as an indicator of poor prognosis in HBC. Matrix degradation and laminin recognition are highlighted as potential targets for antimetastatic therapy, and analyses of laminin attachment and the matrix metalloproteinase (MMP) family in HBC cell lines are summarized. Matrigel-based assays have proved useful in the study of the molecular mechanisms of basement membrane invasiveness, their regulation in HBC cells, and their potential as targets for antimetastatic therapy.

An integrated modeling and risk analysis tool for mine water interactions

Poor mine water management can lead to corporate, environmental and social risks. These risks become more pronounced as mining operations move into areas of water scarcity and/or increase climatic variability while also managing increased demand, lower ore grades and increased strip ratios. Therefore, it is vital that mine sites better understand these risks in order to implement management practices to address them. Systems models provide an effective approach to understand complex networks, particularly across multiple scales. Previous work has represented mine water interactions using systems model on a mine site scale. Here, we expand on that work by present an integrated tool that uses a systems modeling approach to represent mine water interactions on a site and regional scale and then analyses the risks associated with events stemming from those interactions. A case study is presented to represent three indicative corporate, environmental and social risks associated with a mine site that exists in a water scarce region. The tool is generic and flexible, and can be used in many scenarios to provide significant potential utility to the mining industry.

Application of XRF and correspondence analysis to provenance studies of coastal and inland archaeological pottery from the Mngeni river area, South Africa

XRF spectrometry was applied to provenance studies of Iron Age pottery specimens that originated from the Mngeni river area in South Africa. Ten transition metals (Sc to Zn) mere determined in 107 potsherds, excavated from four different sites. The data were subjected to a computerized mathematical technique (correspondence analysis), which was used to group the samples according to the similarity of their elemental distributions. The groupings were interpreted in terms of social or cultural interaction between the sites. (C) 1997 by John Wiley & Sons, Ltd.

Factors influencing robustness and effectiveness of conditional random fields in active learning frameworks

Active learning approaches reduce the annotation cost required by traditional supervised approaches to reach the same effectiveness by actively selecting informative instances during the learning phase. However, effectiveness and robustness of the learnt models are influenced by a number of factors. In this paper we investigate the factors that affect the effectiveness, more specifically in terms of stability and robustness, of active learning models built using conditional random fields (CRFs) for information extraction applications. Stability, defined as a small variation of performance when small variation of the training data or a small variation of the parameters occur, is a major issue for machine learning models, but even more so in the active learning framework which aims to minimise the amount of training data required. The factors we investigate are a) the choice of incremental vs. standard active learning, b) the feature set used as a representation of the text (i.e., morphological features, syntactic features, or semantic features) and c) Gaussian prior variance as one of the important CRFs parameters. Our empirical findings show that incremental learning and the Gaussian prior variance lead to more stable and robust models across iterations. Our study also demonstrates that orthographical, morphological and contextual features as a group of basic features play an important role in learning effective models across all iterations.

Brighter lights on Capital Hill? Speculations upon federal framing and policy analysis of Indigenous issues

Postwar Australian social policy has occurred within neoliberal, social-conservative and social democratic ideational frameworks. Recent perceptions vary from concern about high levels of public spending, through disquiet about cultural change, to fear that government inaction is ignoring community needs and creating fractious and unhealthy social conditions. this paper examines these alternate ideological influences as they could affect Indigenous Australians with a focus on the values and approaches that might lead logically to desirable outcomes. effective policy requires clarity and compatibility between government thinking and the social values of Indigenous people. At issue is how the objectives of policy for Indigenous citizens might be determined.

Lifecycle optimization and tradeoff analysis among competing objectives for a road corridor

While the philosophical motivation behind Civil Infrastructure Management Systems is to achieve optimal level of service at a minimum cost, the allocation of scarce resources among competing alternatives is still a matter of debate. It appears to be widely accepted that results from tradeoff analysis can be measured by the degree of accomplishment of the objectives. Road management systems not only deal with different asset types but also with conflicting objectives. This paper presents a case study of lifecycle optimization with tradeoff analysis for a road corridor in New Brunswick. Objectives of the study included condition of bridge and roads and road safety. A road safety index was created based on potential for improvement. Road condition was based on roughness, rutting and cracking. Initial results show lack of sustainability in bridge performance. Therefore, bridges where broken by components: deck, superstructure and substructure. Visual inspections, in addition to construction age of each bridge, were combined to generate a surrogate apparent age. Two life cycle analysis were conducted; one aimed to minimize overall cost while achieving sustainable results and another one purely for optimization. -used to identify required levels of budget. Such analyses were used to identify the minimum required budget and to demonstrate that with the same amount of money it was possible to achieve better levels of performance. Dominance and performance driven criteria were combined to identify and select an optimal result. It was found that achievement of optimally sustained results is conditioned by the availability of treatments for all asset classes at across their life spans. For the case study a disaggregated bridge condition index was introduced to the original algorithm to attempt to achieve sustainability in all bridges components, however lack of early stage treatments for substructures produce declining trends for such a component.

The long-term health consequences of child physical abuse, emotional abuse, and neglect : a systematic review and meta-analysis

Background Child sexual abuse is considered a modifiable risk factor for mental disorders across the life course. However the long-term consequences of other forms of child maltreatment have not yet been systematically examined. The aim of this study was to summarise the evidence relating to the possible relationship between child physical abuse, emotional abuse, and neglect, and subsequent mental and physical health outcomes. Methods and Findings A systematic review was conducted using the Medline, EMBASE, and PsycINFO electronic databases up to 26 June 2012. Published cohort, cross-sectional, and case-control studies that examined non-sexual child maltreatment as a risk factor for loss of health were included. All meta-analyses were based on quality-effects models. Out of 285 articles assessed for eligibility, 124 studies satisfied the pre-determined inclusion criteria for meta-analysis. Statistically significant associations were observed between physical abuse, emotional abuse, and neglect and depressive disorders (physical abuse [odds ratio (OR) = 1.54; 95% CI 1.16–2.04], emotional abuse [OR = 3.06; 95% CI 2.43–3.85], and neglect [OR = 2.11; 95% CI 1.61–2.77]); drug use (physical abuse [OR = 1.92; 95% CI 1.67–2.20], emotional abuse [OR = 1.41; 95% CI 1.11–1.79], and neglect [OR = 1.36; 95% CI 1.21–1.54]); suicide attempts (physical abuse [OR = 3.40; 95% CI 2.17–5.32], emotional abuse [OR = 3.37; 95% CI 2.44–4.67], and neglect [OR = 1.95; 95% CI 1.13–3.37]); and sexually transmitted infections and risky sexual behaviour (physical abuse [OR = 1.78; 95% CI 1.50–2.10], emotional abuse [OR = 1.75; 95% CI 1.49–2.04], and neglect [OR = 1.57; 95% CI 1.39–1.78]). Evidence for causality was assessed using Bradford Hill criteria. While suggestive evidence exists for a relationship between maltreatment and chronic diseases and lifestyle risk factors, more research is required to confirm these relationships. Conclusions This overview of the evidence suggests a causal relationship between non-sexual child maltreatment and a range of mental disorders, drug use, suicide attempts, sexually transmitted infections, and risky sexual behaviour. All forms of child maltreatment should be considered important risks to health with a sizeable impact on major contributors to the burden of disease in all parts of the world. The awareness of the serious long-term consequences of child maltreatment should encourage better identification of those at risk and the development of effective interventions to protect children from violence.

The role of human factors in led outdoor activity accidents: literature review and exploratory analysis

While the exact rate of incidence is unknown (due to the paucity of exposure data), it is acknowledged that safety compromising accidents and incidents occur in the led outdoor activity domain, and that they represent an important issue. Despite this, compared to other safety critical domains, very little is currently known about the key causal factors involved in such accidents and incidents. This report presents the findings derived from a review of the literature, the aim of which was to identify the Human Factors-related issues involved in accidents and incidents occurring in this area. In addition, to demonstrate the utility of systems-based, theoretically underpinned accident analysis methodologies for identifying the systemic and human contribution to accidents and incidents occurring in the led outdoor activity domain, three case-study accidents were analysed using two such approaches. In conclusion, the review identified a range of causal factors cited in the literature; however, it was noted that the majority of the research undertaken to date lacks theoretical underpinning and focuses mainly on instructor or activity leader causal factors, as opposed to the wider system failures involved. The accident analysis presented highlighted the utility of systems-based, theoretically underpinned accident analysis methodologies for analysing and learning from accidents and incidents in the led outdoor activity sector. In closing, the need for further research in the area is articulated, in particular focussing on the development of standardised and universally accepted accident and incident reporting systems and databases, the development of data driven, theoretically underpinned causal factor taxonomies, and the development and application of systems-based accident analysis methodologies.

Regional aviation and economic growth : cointegration and causality analysis in Australia

Air transport is a critical link to regional, rural and remote communities in Australia. Air services provide important economic and social benefits but very little research has been done on assessing the value of regional aviation. This research provides the first empirical evidence that there is short and long run causality between regional aviation and economic growth. The authors analysed 88 regional airports in Australia over a period of 1985–86 to 2010–11 to determine the catalytic impacts of regional air transport on regional economic growth. The analysis was conducted using annual data related to total airport passenger movements – for the level of airport activity, and real aggregate taxable income – to represent economic growth. A significant bi-directional relationship was established: airports have an impact on regional economic growth and the economy directly impacts regional air transport. The economic significance of regional air transport confirms the importance of the airport as infrastructure for regional councils and the need for them to maintain and develop local airports. Funding should be targeted at airports directly to support regional development.