The development of Lactococcus lactis as an antimicrobial agent

Non-pathogenic lactic acid bacteria are economically important Gram-positive bacteria used extensively in the food industry. Due to their “generally regarded as safe” status, certain species from the genera Lactobacillus and Lactococcus are also considered desirable as candidates for the production and secretion of recombinant proteins, particular those with therapeutic applications. The hypothesis examined by this thesis is that Lactococcus lactis can be modified to be an effective antimicrobial agent. Therefore, the aims of this thesis were to investigate the optimisation of the expression, secretion and/or activities of potential heterologous antimicrobial proteins by the model lactic acid bacterium, Lactococcus lactis subsp. cremoris MG1363. L. lactis strains were engineered to express and secrete the recombinant CyuC surface protein from Lactobacillus reuteri BR11, and a fusion protein consisting of CyuC and lysostaphin using the Sep promoter and secretion signal. CyuC has been characterised as a cystine-binding protein, but has also been demonstrated to have fibronectin binding activity. Lysostaphin is a bacteriolytic enzyme with specific activity against the Gram-positive pathogen, Staphylococcus aureus. These modified L. lactis strains were then investigated to see if they had the ability to inhibit the adhesion of S. aureus to host extracellular matrix (ECM) proteins. It was observed that the cell extracts of the L. lactis strain with the vector only (pGhost9:ISS1) was able to inhibit the adhesion of S. aureus to fibronectin, whilst the cell extracts of the L. lactis strain expressing lysostaphin was able to inhibit adhesion to keratin. Finally, this thesis has identified specific lactococcal genes that affect the secretion of lysostaphin through the use of random transposon mutagenesis. Ten mutants with higher lysostaphin activity contained insertions in four different genes encoding: (i) an uncharacterised putative transmembrane protein (llmg_0609), (ii) an enzyme catalysing the first step in peptidoglycan biosynthesis (murA2), (iii) a homolog of the oxidative defence regulator (trmA), and (iv) an uncharacterised putative enzyme involved in ubiquinone biosynthesis (llmg_2148). The higher lysostaphin activity observed in these mutants was found to be due to higher amounts of lysostaphin being secreted. The findings of this thesis contribute to the development of this organism as an antimicrobial agent and also to our understanding of L. lactis genetics.

Safety in occupational driving : development of a driver behaviour scale for the workplace context

International statistics indicate that occupational, or work-related driving, crashes are the most common cause of workplace injury, death, and absence from work. The majority of research examining unsafe driver behavior in the workplace has relied on general road safety questionnaires. However, past research has failed to consider the organizational context in the use of these questionnaires, and as such, there is ambiguity in the dimensions constituting occupational driving. Using a theoretical model developed by Hockey (1993, 1997), this article proposes and validates a new scale of occupational driver behavior. This scale incorporates four dimensions of driver behavior that are influenced by demanding workplace conditions; speeding, rule violation, inattention, and driving while tired. Following a content validation process, three samples of occupational drivers in Australia were used to assess the scale. Data from the first sample (n=145) were used to reduce the number of scale items and provide an assessment of the factorial validity of the scale. Data from the second sample (n=645) were then used to confirm the factor structure and psychometric properties of the scale including reliability and construct validity. Finally, data from the third sample (n=248) were used to establish criterion validity. The results indicated that the scale is a reliable and valid measure of occupational driver behavior.

Development of a 3-dimensional human skin equivalent wound model for investigating novel wound healing therapies

Numerous difficulties are associated with the conduct of preclinical studies related to skin and wound repair. Use of small animal models such as rodents is not optimal because of their physiological differences to human skin and mode of wound healing. Although pigs have previously been used because of their human-like mode of healing, the expense and logistics related to their use also renders them suboptimal. In view of this, alternatives are urgently required to advance the field. The experiments reported herein were aimed at developing and validating a simple, reproducible, three-dimensional ex vivo de-epidermised dermis human skin equivalent wound model for the preclinical evaluation of novel wound therapies. Having established that the human skin equivalent wound model does in fact “heal," we tested the effect of two novel wound healing therapies. We also examined the utility of the model for studies exploring the mechanisms underpinning these therapies. Taken together the data demonstrate that these new models will have wide-spread application for the generation of fundamental new information on wound healing processes and also hold potential in facilitating preclinical optimization of dosage, duration of therapies, and treatment strategies prior to clinical trials.

A comparative knowledge-based urban development analysis : Vancouver, Melbourne and Manchester vs. Boston

Purpose - The purpose of this paper is to introduce a knowledge-based urban development assessment framework, which has been constructed in order to evaluate and assist in the (re)formulation of local and regional policy frameworks and applications necessary in knowledge city transformations. Design/methodology/approach - The research reported in this paper follows a methodological approach that includes a thorough review of the literature, development of an assessment framework in order to inform policy-making by accurately evaluating knowledge-based development levels of cities, and application of this framework in a comparative study - Boston, Vancouver, Melbourne and Manchester. Originality/value - The paper, with its assessment framework, demonstrates an innovative way of examining the knowledge-based development capacity of cities by scrutinising their economic, socio-cultural, enviro-urban and institutional development mechanisms and capabilities. Practical implications - The paper introduces a framework developed to assess the knowledge-based development levels of cities; presents some of the generic indicators used to evaluate knowledge-based development performance of cities; demonstrates how a city can benchmark its development level against that of other cities, and; provides insights for achieving a more sustainable and knowledge-based development.

Developing managers as researchers using a learning cohort approach

This paper describes the design process and curriculum for a learning cohort of eight managers who came from public and private providers of vocational education and training. While the authors found no discussion on developing research knowledge and skills of managers using learning cohorts, the general learning cohorts literature provided a number of recommendations for learning cohort design. The initial stages of the learning cohort were evaluated. The results highlighted the importance of clarifying the psychological contract and its use in self-selection, supported the recommendations in the literature of the significance of the careful design and implementation of an initial residential workshop and also found support for further residential workshops of a similar design. The attendance of the cohort members in tow faculty wide core research units drew mixed comments.

The development and validation of two complementary measures of drivers' hazard perception ability

Hazard perception in driving involves a number of different processes. This paper reports the development of two measures designed to separate these processes. A Hazard Perception Test was developed to measure how quickly drivers could anticipate hazards overall, incorporating detection, trajectory prediction, and hazard classification judgements. A Hazard Change Detection Task was developed to measure how quickly drivers can detect a hazard in a static image regardless of whether they consider it hazardous or not. For the Hazard Perception Test, young novices were slower than mid-age experienced drivers, consistent with differences in crash risk, and test performance correlated with scores in pre-existing Hazard Perception Tests. For drivers aged 65 and over, scores on the Hazard Perception Test declined with age and correlated with both contrast sensitivity and a Useful Field of View measure. For the Hazard Change Detection Task, novices responded quicker than the experienced drivers, contrary to crash risk trends, and test performance did not correlate with measures of overall hazard perception. However for drivers aged 65 and over, test performance declined with age and correlated with both hazard perception and Useful Field of View. Overall we concluded that there was support for the validity of the Hazard Perception Test for all ages but the Hazard Change Detection Task might only be appropriate for use with older drivers.

Executive function effects and numerical development in children : behavioural and ERP evidence from a numerical Stroop paradigm

Most research on numerical development in children is behavioural, focusing on accuracy and response time in different problem formats. However, Temple and Posner (1998) used ERPs and the numerical distance task with 5-year-olds to show that the development of numerical representations is difficult to disentangle from the development of the executive components of response organization and execution. Here we use the numerical Stroop paradigm (NSP) and ERPs to study possible executive interference in numerical processing tasks in 6–8-year-old children. In the NSP, the numerical magnitude of the digits is task-relevant and the physical size of the digits is task-irrelevant. We show that younger children are highly susceptible to interference from irrelevant physical information such as digit size, but that access to the numerical representation is almost as fast in young children as in adults. We argue that the developmental trajectories for executive function and numerical processing may act together to determine numerical development in young children.

Autonomous underwater vehicles: development and implementation of time and energy efficient trajectories

Autonomous underwater vehicles (AUVs) are increasingly used, both in military and civilian applications. These vehicles are limited mainly by the intelligence we give them and the life of their batteries. Research is active to extend vehicle autonomy in both aspects. Our intent is to give the vehicle the ability to adapt its behavior under different mission scenarios (emergency maneuvers versus long duration monitoring). This involves a search for optimal trajectories minimizing time, energy or a combination of both. Despite some success stories in AUV control, optimal control is still a very underdeveloped area. Adaptive control research has contributed to cost minimization problems, but vehicle design has been the driving force for advancement in optimal control research. We look to advance the development of optimal control theory by expanding the motions along which AUVs travel. Traditionally, AUVs have taken the role of performing the long data gathering mission in the open ocean with little to no interaction with their surroundings, MacIver et al. (2004). The AUV is used to find the shipwreck, and the remotely operated vehicle (ROV) handles the exploration up close. AUV mission profiles of this sort are best suited through the use of a torpedo shaped AUV, Bertram and Alvarez (2006), since straight lines and minimal (0 deg - 30 deg) angular displacements are all that are necessary to perform the transects and grid lines for these applications. However, the torpedo shape AUV lacks the ability to perform low-speed maneuvers in cluttered environments, such as autonomous exploration close to the seabed and around obstacles, MacIver et al. (2004). Thus, we consider an agile vehicle capable of movement in six degrees of freedom without any preference of direction.

The airport city : a new business model for airport development

The airport city concept has been embraced by many airports of different scales and in varied ways around the world. Airports everywhere have diversified their landside revenues with non-aviation commercial and industrial development in order to increase revenues and spread risk in the notoriously volatile aviation market. As intermodal hubs in a connected, globalised world, airports have evolved from transportation nodes into multi-faceted business enterprises. They have assumed a critical role as ‘transactional’ spaces in the global economy (Gottdiener 2001).

Contexts, hybrids and network governance : a comparison of three case-studies in infrastructure governance

While hybrid governance arrangements have been a major element of organisational architecture for some time, the contemporary operating environment has brought to the fore new conditions and expectations for the governance of entities that span conventional public sector departments, private firms and community organisations or groups. These conditions have resulted in a broader array of mixed governance configurations including Public Private Partnerships, alliances, and formal and informal collaborations. In some such arrangements, market based or ‘complete’ contractual relationships have been introduced to replace or supplement existing traditional ‘hierarchical’ and/or newer relational ‘network-oriented’ institutional associations. While there has been a greater reliance on collaborative or relational contracts as an underpinning institutional model, other modes of hierarchy and market may remain in operation. The success of these emergent hybrid forms has been mixed. There are examples of hybrids that have been well adopted, achieving the desired goals of efficiency, effectiveness and financial accountability; while others have experienced implementation problems which have undermined their results. This paper postulates that the cultural and institutional context within which hybrids operate may contribute to the implementation processes employed and the level of success attained. The paper explores hybrid arrangements through three cases of the use of inter-organisational arrangements in three different national contexts. Distilling the various elements of hybrids and the impact of institutional context will provide important insights for those charged with the responsibility for the formation and key infrastructure and public value development.

Attachment in relation to affect regulation and interpersonal functioning among substance use disorder inpatients

Attachment theory has been conceptualised as an affect regulation theory, proposing that attachment is associated with the expression and recognition of emotions as well as interpersonal functioning. Previous research has reported affect regulation difficulties in substance use disorders and addiction has been considered an attachment disorder. However, scarce empirical research exists on the relationship of attachment in relation to affect regulation and interpersonal functioning in those with substance use problems. Thus, the objective of the present study was to investigate potential associations between attachment, negative mood regulation (NMR) expectancies, fear of intimacy and self-differentiation in substance abusers. The revised adult attachment scale (RAAS), the NMR expectancies scale, the fear of intimacy scale and the differentiation of self inventory were administered to a sample of 100 substance use disorder inpatients. Attachment accounted for significant variance in NMR expectancies and was also a strong predictor of fear of intimacy. The predictive utility of attachment also extended to self-differentiation, suggesting that attachment was strongly related to overall self-differentiation score, Emotional reactivity, Emotional cut-off and I position. These findings support attachment theory suggesting that attachment is associated with and predicts affect regulation abilities and difficulties in interpersonal functioning in a sample of substance use disorder inpatients. The inclusion and assessment of attachment appears to be important in the development of treatment programmes for substance abusing individuals.

Use of acoustic emission technique for structural health monitoring of bridges

Bridges are valuable assets of every nation. They deteriorate with age and often are subjected to additional loads or different load patterns than originally designed for. These changes in loads can cause localized distress and may result in bridge failure if not corrected in time. Early detection of damage and appropriate retrofitting will aid in preventing bridge failures. Large amounts of money are spent in bridge maintenance all around the world. A need exists for a reliable technology capable of monitoring the structural health of bridges, thereby ensuring they operate safely and efficiently during the whole intended lives. Monitoring of bridges has been traditionally done by means of visual inspection. Visual inspection alone is not capable of locating and identifying all signs of damage, hence a variety of structural health monitoring (SHM) techniques is used regularly nowadays to monitor performance and to assess condition of bridges for early damage detection. Acoustic emission (AE) is one technique that is finding an increasing use in SHM applications of bridges all around the world. The chapter starts with a brief introduction to structural health monitoring and techniques commonly used for monitoring purposes. Acoustic emission technique, wave nature of AE phenomenon, previous applications and limitations and challenges in the use as a SHM technique are also discussed. Scope of the project and work carried out will be explained, followed by some recommendations of work planned in future.

The use of supply chain management to reduce delays : Malaysian public sector construction projects

Construction delays are a critical problem for Malaysian public sector projects. These delays have been blamed mainly on inefficient traditional construction practices that continue to dominate the current industry. This paper reports the progress to date of a Ph.D. research project aimed at developing a framework to utilize Supply Chain Management (SCM) tools to improve the time performance of Malaysian Government projects. The potential of SCM has been identified for public sector governance and its use in Malaysia is now being considered within the strategy of the Malaysian Construction Industry Master Plan (2006-2015). Encouraged by success in the UK, there is a cautious optimism concerning the successful application of SCM in Malaysia. This paper considers delay as a problem in Malaysian public sector projects, establishes the need to embrace SCM and then elucidates the need and strategies for the development of a delay reduction framework. A literature review, survey mechanism and structured interview schedule will be undertaken to achieve the research objectives. The final research outcome will be a framework that addresses root delay contributors (“pathogens”) and applies SCM tools for their mitigation.

Practical introduction to development of sage applications

Throughout this workshop session we have looked at various configurations of Sage as well as using the Sage UI to run Sage applications (e.g. the image viewer). More advanced usage of Sage has been demonstrated using a Sage compatible version of Paraview highlighting the potential of parallel rendering. The aim of this tutorial session is to give a practical introduction to developing visual content for a tiled display using the Sage libraries. After completing this tutorial you should have the basic tools required to develop your own custom Sage applications. This tutorial is designed for software developers and intermediate programming knowledge is assumed, along with some introductory OpenGL . You will be required to write small portions of C/C++ code to complete this worksheet. However if you do not feel comfortable writing code (or have never written in C or C++), we will be on hand throughout this session so feel free to ask for some help. We have a number of machines in this lab running a VNC client to a virtual machine running Fedora 12. You should all be able to log in with the username “escience”, and password “escience10”. Some of the commands in this worksheet require you to run them as the root user, so note the password as you may need to use it a few times. If you need to access the Internet, then use the username “qpsf01”, password “escience10”

Essays on irrigation development, farm production and unaccounted costs : theory and empirical evidence

A review of the literature related to issues involved in irrigation induced agricultural development (IIAD) reveals that: (1) the magnitude, sensitivity and distribution of social welfare of IIAD is not fully analysed; (2) the impacts of excessive pesticide use on farmers’ health are not adequately explained; (3) no analysis estimates the relationship between farm level efficiency and overuse of agro-chemical inputs under imperfect markets; and (4) the method of incorporating groundwater extraction costs is misleading. This PhD thesis investigates these issues by using primary data, along with secondary data from Sri Lanka. The overall findings of the thesis can be summarised as follows. First, the thesis demonstrates that Sri Lanka has gained a positive welfare change as a result of introducing new irrigation technology. The change in the consumer surplus is Rs.48,236 million, while the change in the producer surplus is Rs. 14,274 millions between 1970 and 2006. The results also show that the long run benefits and costs of IIAD depend critically on the magnitude of the expansion of the irrigated area, as well as the competition faced by traditional farmers (agricultural crowding out effects). The traditional sector’s ability to compete with the modern sector depends on productivity improvements, reducing production costs and future structural changes (spillover effects). Second, the thesis findings on pesticides used for agriculture show that, on average, a farmer incurs a cost of approximately Rs. 590 to 800 per month during a typical cultivation period due to exposure to pesticides. It is shown that the value of average loss in earnings per farmer for the ‘hospitalised’ sample is Rs. 475 per month, while it is approximately Rs. 345 per month for the ‘general’ farmers group during a typical cultivation season. However, the average willingness to pay (WTP) to avoid exposure to pesticides is approximately Rs. 950 and Rs. 620 for ‘hospitalised’ and ‘general’ farmers’ samples respectively. The estimated percentage contribution for WTP due to health costs, lost earnings, mitigating expenditure, and disutility are 29, 50, 5 and 16 per cent respectively for hospitalised farmers, while they are 32, 55, 8 and 5 per cent respectively for ‘general’ farmers. It is also shown that given market imperfections for most agricultural inputs, farmers are overusing pesticides with the expectation of higher future returns. This has led to an increase in inefficiency in farming practices which is not understood by the farmers. Third, it is found that various groundwater depletion studies in the economics literature have provided misleading optimal water extraction quantity levels. This is due to a failure to incorporate all production costs in the relevant models. It is only by incorporating quality changes to quantity deterioration, that it is possible to derive socially optimal levels. Empirical results clearly show that the benefits per hectare per month considering both the avoidance costs of deepening agro-wells by five feet from the existing average, as well as the avoidance costs of maintaining the water salinity level at 1.8 (mmhos/Cm), is approximately Rs. 4,350 for farmers in the Anuradhapura district and Rs. 5,600 for farmers in the Matale district.