Monitoring healing progression and characterizing the mechanical environment in preclinical models for bone tissue engineering

The treatment of large segmental bone defects remains a significant clinical challenge. Due to limitations surrounding the use of bone grafts, tissue-engineered constructs for the repair of large bone defects could offer an alternative. Before translation of any newly developed tissue engineering (TE) approach to the clinic, efficacy of the treatment must be shown in a validated preclinical large animal model. Currently, biomechanical testing, histology, and microcomputed tomography are performed to assess the quality and quantity of the regenerated bone. However, in vivo monitoring of the progression of healing is seldom performed, which could reveal important information regarding time to restoration of mechanical function and acceleration of regeneration. Furthermore, since the mechanical environment is known to influence bone regeneration, and limb loading of the animals can poorly be controlled, characterizing activity and load history could provide the ability to explain variability in the acquired data sets and potentially outliers based on abnormal loading. Many approaches have been devised to monitor the progression of healing and characterize the mechanical environment in fracture healing studies. In this article, we review previous methods and share results of recent work of our group toward developing and implementing a comprehensive biomechanical monitoring system to study bone regeneration in preclinical TE studies.

25th Anniversary of the Linked Life Insurance Commemorative Medal

Obverse: Inscription. Reverse: The life insurance emblem, a house with family inside, shekel coin on the roof of the house.

Plasma polymer and biomolecule modification of 3-D scaffolds for tissue engineering

Plasma polymerization was used to coat a melt electrospun polycaprolactone scaffold to improve cell attachment and organization. Plasma polymerization was performed using an amine containing monomer, allylamine, which then allowed for the subsequent immobilization of biomolecules i.e. heparin and fibroblast growth factor-2. The stability of the plasma polymerized amine-coating was demonstrated by X-ray photoelectron spectroscopy analysis and imaging time-of-flight secondary ion mass spectrometry revealed that a uniform plasma amine-coating was deposited throughout the scaffold. Based upon comparison with controls it was evident that the combination scaffold aided cell ingress and the formation of distinct fibroblast and keratinocyte layers.

What does built environment research have to do with risk mitigation, resilience and disaster recovery?

One could argue that the nature of our housing stock is a key determining factor in the ability of our citizens to manage risk, be resilient to various natural and human events, and to recover from these events. Recent research has been examining current challenges posed by our housing stock and exploring potential solutions from a range of perspectives. The aim of this paper is to discuss key findings from recent built environment research in Australia to initiate cross-sectorial discussion and debate about the implications and opportunities for other sectors such as emergency management and insurance. Three recent building research projects are discussed: - Heat waves The impact of heat waves on houses and occupants, and proposed changes to building regulations, air conditioning standards and building design, to reduce risks associated with heat waves. - Net zero energy homes Exploration of the potential benefits of a strategic optimization of building quality, energy and water efficiency, and household or community level distributed energy and water services for disaster management and recovery. - Building information Mapping of the flow of information about residential buildings, and the potential for national or regional building files (in a similar manner to personal medical records) to assist all parties to make more informed decisions that impact on housing sustainability and community resilience. The paper discusses how sustainability, environmental performance and resilience are inter-related, and can be supported by building files. It concludes with a call for increased cross-sectorial collaboration to explore opportunities for a whole-of-systems approach to our built environment that addresses a range of economic and environmental challenges as well as disaster and emergency management.

Accounting for risk: The advent of capped conveyancing title insurance

Title insurance companies originating from America, have, in the past 15 years become part of the Australian conveyancing landscape. However for most residential freehold owners, their activities would be a mystery. A purchaser does not routinely obtain title insurance, with the companies presently focussing on servicing the mortgagee sector. While the lack of penetration in the residential purchaser market may be attributed to the consumer’s lack of knowledge, evidence from Ontario and New Zealand illustrates that title insurance is likely to become an additional cost in the conveyancing process in Australia. In this article we highlight the reasons why, and demonstrate how title insurers have, by working with the legal profession been able to subtly move the risk of responsibility for compensation for loss, (at least in the first instance) from the state to the insurer, but with the added benefit for the state and the conveyancing agents that the cost of the insurance is ultimately borne by the consumer. In New Zealand this development is being accelerated by the introduction of capped conveyancing title insurance. Whether title insurance will become part of the conveyancing process is no longer the relevant question for Australia, (it undoubtedly will), but the unknown issue is just how title insurance companies will work with conveyancing agents to infiltrate the market, and what response this infiltration will have in terms of the state’s view as to their continued role in the provision of assurance. We suggest that developments from New Zealand in relation to capped conveyancing insurance are likely to be replicated in Australia in the near future, and that the state’s role in providing an assurance fund will continue, though the state may seek to expand the areas in which the right to compensation is restricted.

Auditing design and innovation programmes within a multinational insurance provider

The financial services industry is key to the success of the Australian economy, however even established businesses need to continually push to innovate. At present, design is becoming widely accepted as means to innovate and many companies currently have design and innovation programmes. The goal of these programmes is to foster a design capability amongst staff members. However, little is known as to the impact of the innovation capabilities such design programmes claim to instil. Therefore, this paper seeks to investigate how a financial services firm is currently attempting to build a design capability amongst staff members. Using a case study approach the research team audited four design capability programmes currently being deployed by the case firm. The study highlights the strengths and weaknesses of current efforts by the firm and current barriers to achieving design capability. The implications of this paper include key insights for both academics and industry.

Milking Diatoms for Sustainable Energy: Biochemical Engineering versus Gasoline-Secreting Diatom Solar Panels

In the face of increasing CO2 emissions from conventional energy (gasoline), and the anticipated scarcity of Crude oil, a worldwide effort is underway for cost-effective renewable alternative energy sources. Here, we review a simple line of reasoning: (a) geologists claim that Much crude oil comes from diatoms; (b) diatoms do indeed make oil; (c) agriculturists Claim that diatoms could make 10-200 times as much oil per hectare as oil seeds; and (d) therefore, sustainable energy could be made from diatoms. In this communication, we propose ways of harvesting oil from diatoms, using biochemical engineering and also a new solar panel approach that utilizes genomically modifiable aspects of diatom biology, offering the prospect of ``milking'' diatoms for Sustainable energy by altering them to actively secrete oil products. Secretion by and milking of diatoms may provide a way around the puzzle of how to make algae that both grow quickly and have a very high oil content.

Directors and officers insurance – Recent critical issues in Australia and New Zealand

Directors and Officers Liability Insurance (“D&O insurance”) has grown and evolved rapidly over the past 80 years to assume an important position in most corporations’ corporate governance and risk management strategies. This article focuses upon certain topical matters of particular concern to directors and officers including the availability of defence costs where a D&O policy is subject to a statutory charge; the commercial desirability of stand-alone “A-side” coverage, being the cover provided directly to directors and officers for loss resulting from claims made against them for wrongful acts; the impact of fraud and/or dishonesty upon D&O cover; and disclosure of the nature and extent of D&O cover to the directors and officers themselves and to third parties – in the latter case such access frequently being necessary to determine the economic viability of pursuing a proposed action against a company and its directors and officers.

Engineering the pentose phosphate pathway of Saccharomyces cerevisiae for production of ethanol and xylitol

The baker s yeast Saccharomyces cerevisiae has a long tradition in alcohol production from D-glucose of e.g. starch. However, without genetic modifications it is unable to utilise the 5-carbon sugars D-xylose and L arabinose present in plant biomass. In this study, one key metabolic step of the catabolic D-xylose pathway in recombinant D-xylose-utilising S. cerevisiae strains was studied. This step, carried out by xylulokinase (XK), was shown to be rate-limiting, because overexpression of the xylulokinase-encoding gene XKS1 increased both the specific ethanol production rate and the yield from D xylose. In addition, less of the unwanted side product xylitol was produced. Recombinant D-xylose-utilizing S. cerevisiae strains have been constructed by expressing the genes coding for the first two enzymes of the pathway, D-xylose reductase (XR) and xylitol dehydrogenase (XDH) from the D-xylose-utilising yeast Pichia stipitis. In this study, the ability of endogenous genes of S. cerevisiae to enable D-xylose utilisation was evaluated. Overexpression of the GRE3 gene coding for an unspecific aldose reductase and the ScXYL2 gene coding for a xylitol dehydrogenase homologue enabled growth on D-xylose in aerobic conditions. However, the strain with GRE3 and ScXYL2 had a lower growth rate and accumulated more xylitol compared to the strain with the corresponding enzymes from P. stipitis. Use of the strictly NADPH-dependent Gre3p instead of the P. stipitis XR able to utilise both NADH and NADPH leads to a more severe redox imbalance. In a S. cerevisiae strain not engineered for D-xylose utilisation the presence of D-xylose increased xylitol dehydrogenase activity and the expression of the genes SOR1 or SOR2 coding for sorbitol dehydrogenase. Thus, D-xylose utilisation by S. cerevisiae with activities encoded by ScXYL2 or possibly SOR1 or SOR2, and GRE3 is feasible, but requires efficient redox balance engineering. Compared to D-xylose, D-glucose is a cheap and readily available substrate and thus an attractive alternative for xylitol manufacture. In this study, the pentose phosphate pathway (PPP) of S. cerevisiae was engineered for production of xylitol from D-glucose. Xylitol was formed from D-xylulose 5-phosphate in strains lacking transketolase activity and expressing the gene coding for XDH from P. stipitis. In addition to xylitol, ribitol, D-ribose and D-ribulose were also formed. Deletion of the xylulokinase-encoding gene increased xylitol production, whereas the expression of DOG1 coding for sugar phosphate phosphatase increased ribitol, D-ribose and D-ribulose production. Strains lacking phosphoglucose isomerase (Pgi1p) activity were shown to produce 5 carbon compounds through PPP when DOG1 was overexpressed. Expression of genes encoding glyceraldehyde 3-phosphate dehydrogenase of Bacillus subtilis, GapB, or NAD-dependent glutamate dehydrogenase Gdh2p of S. cerevisiae, altered the cellular redox balance and enhanced growth of pgi1 strains on D glucose, but co-expression with DOG1 reduced growth on higher D-glucose concentrations. Strains lacking both transketolase and phosphoglucose isomerase activities tolerated only low D-glucose concentrations, but the yield of 5-carbon sugars and sugar alcohols on D-glucose was about 50% (w/w).

An engineering or behavioural approach? A study into employee’s perceptions regarding the effectiveness of occupational road safety initiatives

Background and Aims Considerable variation has been documented with fleet safety interventions’ abilities to create lasting behavioural change, and research has neglected to consider employees’ perceptions regarding the effectiveness of fleet interventions. This is a critical oversight as employees’ beliefs and acceptance levels (as well as the perceived organisational commitment to safety) can ultimately influence levels of effectiveness, and this study aimed to examine such perceptions in Australian fleet settings. Method 679 employees sourced from four Australian organisations completed a safety climate questionnaire as well as provided perspectives about the effectiveness of 35 different safety initiatives. Results Countermeasures that were perceived as most effective were a mix of human and engineering-based approaches: - (a) purchasing safer vehicles; - (b) investigating serious vehicle incidents, and; - (c) practical driver skills training. In contrast, least effective countermeasures were considered to be: - (a) signing a promise card; - (b) advertising a company’s phone number on the back of cars for complaints and compliments, and; - (c) communicating cost benefits of road safety to employees. No significant differences in employee perceptions were identified based on age, gender, employees’ self-reported crash involvement or employees’ self-reported traffic infringement history. Perceptions of safety climate were identified to be “moderate” but were not linked to self-reported crash or traffic infringement history. However, higher levels of safety climate were positively correlated with perceived effectiveness of some interventions. Conclusion Taken together, employees believed occupational road safety risks could best be managed by the employer by implementing a combination of engineering and human resource initiatives to enhance road safety. This paper will further outline the key findings in regards to practice as well as provide direction for future research.

Mu in vitro transposition applications in protein engineering

Transposons, mobile genetic elements that are ubiquitous in all living organisms have been used as tools in molecular biology for decades. They have the ability to move into discrete DNA locations with no apparent homology to the target site. The utility of transposons as molecular tools is based on their ability to integrate into various DNA sequences efficiently, producing extensive mutant clone libraries that can be used in various molecular biology applications. Bacteriophage Mu is one of the most useful transposons due to its well-characterized and simple in vitro transposition reaction. This study establishes the properties of the Mu in vitro transposition system as a versatile multipurpose tool in molecular biology. In addition, this study describes Mu-based applications for engineering proteins by random insertional transposon mutagenesis in order to study structure-function relationships in proteins. We initially characterized the properties of the minimal Mu in vitro transposition system. We showed that the Mu transposition system works efficiently and accurately and produces insertions into a wide spectrum of target sites in different DNA molecules. Then, we developed a pentapeptide insertion mutagenesis strategy for inserting random five amino acid cassettes into proteins. These protein variants can be used especially for screening important sites for protein-protein interactions. Also, the system may produce temperature-sensitive variants of the protein of interest. Furthermore, we developed an efficient screening system for high-resolution mapping of protein-protein interfaces with the pentapeptide insertion mutagenesis. This was accomplished by combining the mutagenesis with subsequent yeast two-hybrid screening and PCR-based genetic footprinting. This combination allows the analysis of the whole mutant library en masse, without the need for producing or isolating separate mutant clones, and the protein-protein interfaces can be determined at amino acid accuracy. The system was validated by analysing the interacting region of JFC1 with Rab8A, and we show that the interaction is mediated via the JFC1 Slp homology domain. In addition, we developed a procedure for the production of nested sets of N- and C-terminal deletion variants of proteins with the Mu system. These variants are useful in many functional studies of proteins, especially in mapping regions involved in protein-protein interactions. This methodology was validated by analysing the region in yeast Mso1 involved in an interaction with Sec1. The results of this study show that the Mu in vitro transposition system is versatile for various applicational purposes and can efficiently be adapted to random protein engineering applications for functional studies of proteins.

User centred design for engineering e-government web usability

E-government provides a platform for governments to implement web enabled services that facilitate communication between citizens and the government. However, technology driven design approach and limited understanding of citizens' requirements, have led to a number of critical usability problems on the government websites. Hitherto, there has been no systematic attempt to analyse the way in which theory of User Centred Design (UCD) can contribute to address the usability issues of government websites. This research seeks to fill this gap by synthesising perspectives drawn from the study of User Centred Design and examining them based on the empirical data derived from case study of the Scottish Executive website. The research employs a qualitative approach in the collection and analysis of data. The triangulated analysis of the findings reveals that e-government web designers take commercial development approach and focus only on technical implementations which lead to websites that do not meet citizens' expectations. The research identifies that e-government practitioners can overcome web usability issues by transferring the theory of UCD to practice.