Evaluation of methods for cultivating limbal mesenchymal stromal cells

Background: Mesenchymal stromal cells (MSC) with similar properties to bone marrow derived mesenchymal stromal cells (BM-MSC) have recently been grown from the limbus of the human cornea. We presently contribute to this novel area of research by evaluating methods for culturing human limbal MSC (L-MSC). Methods: Four basic strategies are compared: serum-supplemented medium (10% foetal bovine serum; FBS), standard serum-free medium supplemented with B-27, epidermal growth factor, and fibroblast growth factor 2, or one of two commercial serum-free media including Defined Keratinocyte Serum Free Medium (Invitrogen), and MesenCult-XF (Stem Cell Technologies). The phenotype of resulting cultures was examined using photography, flow cytometry (for CD34, CD45, CD73, CD90, CD105, CD141, CD271), immunocytochemistry (α-sma), differentiation assays (osteogenesis, adipogenesis, chrondrogenesis), and co-culture experiments with human limbal epithelial (HLE) cells. Results: While all techniques supported to varying degrees establishment of cultures, sustained growth and serial propagation was only achieved in 10% FBS medium or MesenCult-XF medium. Cultures established in 10% FBS medium were 70-80% CD34-/CD45-/CD90+/CD73+/CD105+, approximately 25% α-sma+, and displayed multi-potency. Cultures established in MesenCult-XF were >95% CD34-/CD45-/CD90+/CD73+/CD105+, 40% CD141+, rarely expressed α-sma, and displayed multi-potency. L-MSC supported growth of HLE cells, with the largest epithelial islands being observed in the presence of MesenCult-XF-grown L-MSC. All HLE cultures supported by L-MSC widely expressed the progenitor cell marker ∆Np63, along with the corneal differentiation marker cytokeratin 3. Conclusions: We conclude that MesenCult-XF® is a superior culture system for L-MSC, but further studies are required to explore the significance of CD141 expression in these cells.

An investigation of gaseous and particulate emissions from compression ignition engines operated with alternative fuels, injection technologies, and combustion strategies

Whilst the compression ignition (CI) engine exhibits many design advantages relative to its spark ignition engine counterpart; such as: high thermal efficiency, fuel economy and low carbon monoxide and hydrocarbon emissions, the issue of Diesel Particulate Matter (DPM) emissions continues to be an unresolved problem for the CI engine. Primarily, this thesis investigates a range of DPM mitigation strategies such as alternative fuels, injection technologies and combustion strategies conducted with a view to determine their impact on the physico-chemical properties of DPM emissions, and consequently to shed light on their likely human health impacts. Regulated gaseous emissions, Nitric oxide (NO), Carbon monoxide (CO), and Hydrocarbons (HCs), were measured in all experimental campaigns, although the major focus in this research program was on particulate emissions...

Towards an integration of the theory of planned behaviour and cognitive behavioural strategies : an example from a school-based injury prevention programme

Adolescent risk-taking behavior has potentially serious injury consequences and school-based behavior change programs provide potential for reducing such harm. A well-designed program is likely to be theory-based and ecologically valid however it is rare that the operationalisation process of theories is described. The aim of this paper is to outline how the Theory of Planned Behavior and Cognitive Behavioral Therapy informed intervention design in a school setting. Teacher interviews provided insights into strategies that might be implemented within the curriculum and provided detail used to operationalise theory constructs. Benefits and challenges in applying both theories are described with examples from an injury prevention program, Skills for Preventing Injury in Youth.

Agronomic and economic evaluation of irrigation strategies on cotton lint yield in Australia

Cotton is one of the most important irrigated crops in subtropical Australia. In recent years, cotton production has been severely affected by the worst drought in recorded history, with the 2007–08 growing season recording the lowest average cotton yield in 30 years. The use of a crop simulation model to simulate the long-term temporal distribution of cotton yields under different levels of irrigation and the marginal value for each unit of water applied is important in determining the economic feasibility of current irrigation practices. The objectives of this study were to: (i) evaluate the CROPGRO-Cotton simulation model for studying crop growth under deficit irrigation scenarios across ten locations in New South Wales (NSW) and Queensland (Qld); (ii) evaluate agronomic and economic responses to water inputs across the ten locations; and (iii) determine the economically optimal irrigation level. The CROPGRO-Cotton simulation model was evaluated using 2 years of experimental data collected at Kingsthorpe, Qld. The model was further evaluated using data from nine locations between northern NSW and southern Qld. Long-term simulations were based on the prevalent furrowirrigation practice of refilling the soil profile when the plant -available soil water content is<50%. The model closely estimated lint yield for all locations evaluated. Our results showed that the amounts of water needed to maximise profit and maximise yield are different, which has economic and environmental implications. Irrigation needed to maximise profits varied with both agronomic and economic factors, which can be quite variable with season and location. Therefore, better tools and information that consider the agronomic and economic implications of irrigation decisions need to be developed and made available to growers.

Sustainable water provision : challenges, alternative strategies and sources in the ERA of climate change

As a result of rapid urbanisation, population growth, changes in lifestyle, pollution and the impacts of climate change, water provision has become a critical challenge for planners and policy-makers. In the wake of increasingly difficult water provision and drought, the notion that freshwater is a finite and vulnerable resource is increasingly being realised. Many city administrations around the world are struggling to provide water security for their residents to maintain lifestyle and economic growth. This chapter reviews the global challenge of providing freshwater to sustain lifestyles and economic growth, and the contributing challenges of climate change, urbanisation, population growth and problems in rainfall distribution. The chapter proceeds to evaluate major alternatives to current water sources such as conservation, recycling and reclamation, and desalination. Integrated water resource management is briefly looked at to explore its role in complementing water provision. A comparative study on alternative resources is undertaken to evaluate their strengths, weaknesses, opportunities and constraints, and the results are discussed.

Evaluation of the cost-effectiveness of strategies claiming to reduce the risk of surgical site infections following primary total hip arthroplasty

Background Total hip arthroplasty (THA) is a commonly performed procedure and numbers are increasing with ageing populations. One of the most serious complications in THA are surgical site infections (SSIs), caused by pathogens entering the wound during the procedure. SSIs are associated with a substantial burden for health services, increased mortality and reduced functional outcomes in patients. Numerous approaches to preventing these infections exist but there is no gold standard in practice and the cost-effectiveness of alternate strategies is largely unknown. Objectives The aim of this project was to evaluate the cost-effectiveness of strategies claiming to reduce deep surgical site infections following total hip arthroplasty in Australia. The objectives were: 1. Identification of competing strategies or combinations of strategies that are clinically relevant to the control of SSI related to hip arthroplasty 2. Evidence synthesis and pooling of results to assess the volume and quality of evidence claiming to reduce the risk of SSI following total hip arthroplasty 3. Construction of an economic decision model incorporating cost and health outcomes for each of the identified strategies 4. Quantification of the effect of uncertainty in the model 5. Assessment of the value of perfect information among model parameters to inform future data collection Methods The literature relating to SSI in THA was reviewed, in particular to establish definitions of these concepts, understand mechanisms of aetiology and microbiology, risk factors, diagnosis and consequences as well as to give an overview of existing infection prevention measures. Published economic evaluations on this topic were also reviewed and limitations for Australian decision-makers identified. A Markov state-transition model was developed for the Australian context and subsequently validated by clinicians. The model was designed to capture key events related to deep SSI occurring within the first 12 months following primary THA. Relevant infection prevention measures were selected by reviewing clinical guideline recommendations combined with expert elicitation. Strategies selected for evaluation were the routine use of pre-operative antibiotic prophylaxis (AP) versus no use of antibiotic prophylaxis (No AP) or in combination with antibiotic-impregnated cement (AP & ABC) or laminar air operating rooms (AP & LOR). The best available evidence for clinical effect size and utility parameters was harvested from the medical literature using reproducible methods. Queensland hospital data were extracted to inform patients’ transitions between model health states and related costs captured in assigned treatment codes. Costs related to infection prevention were derived from reliable hospital records and expert opinion. Uncertainty of model input parameters was explored in probabilistic sensitivity analyses and scenario analyses and the value of perfect information was estimated. Results The cost-effectiveness analysis was performed from a health services perspective using a hypothetical cohort of 30,000 THA patients aged 65 years. The baseline rate of deep SSI was 0.96% within one year of a primary THA. The routine use of antibiotic prophylaxis (AP) was highly cost-effective and resulted in cost savings of over $1.6m whilst generating an extra 163 QALYs (without consideration of uncertainty). Deterministic and probabilistic analysis (considering uncertainty) identified antibiotic prophylaxis combined with antibiotic-impregnated cement (AP & ABC) to be the most cost-effective strategy. Using AP & ABC generated the highest net monetary benefit (NMB) and an incremental $3.1m NMB compared to only using antibiotic prophylaxis. There was a very low error probability that this strategy might not have the largest NMB (<5%). Not using antibiotic prophylaxis (No AP) or using both antibiotic prophylaxis combined with laminar air operating rooms (AP & LOR) resulted in worse health outcomes and higher costs. Sensitivity analyses showed that the model was sensitive to the initial cohort starting age and the additional costs of ABC but the best strategy did not change, even for extreme values. The cost-effectiveness improved for a higher proportion of cemented primary THAs and higher baseline rates of deep SSI. The value of perfect information indicated that no additional research is required to support the model conclusions. Conclusions Preventing deep SSI with antibiotic prophylaxis and antibiotic-impregnated cement has shown to improve health outcomes among hospitalised patients, save lives and enhance resource allocation. By implementing a more beneficial infection control strategy, scarce health care resources can be used more efficiently to the benefit of all members of society. The results of this project provide Australian policy makers with key information about how to efficiently manage risks of infection in THA.

Mission drift or strategic shift? Group formation strategies within MEPs

Microenterprise development programs (MEPs) have been recognised as a valuable way to help the poor engage in micro-businesses (Green et al., 2006; Vargas, 2000), presenting a way out of poverty (Choudhury et al., 2008; Strier, 2010). Concerns have been raised however, that the benefits of MEPs often don’t reach the extremely poor (Jones et al., 2004; Midgley, 2008; Mosley and Hulme, 1998; Nawaz, 2010; Pritchett, 2006). Balancing reach of these programs with depth is a challenging task. Targeting as many poor people as possible often results in MEPs focusing on the upper or middle poor, overlooking the most challenging group. As such, MEPs have been criticised for mission drift – losing sight of the organisation’s core purpose; assisting those more likely to succeed.

The effect of silicate ions on proliferation, osteogenic differentiation and cell signalling pathways (WNT and SHH) of bone marrow stromal cells

Silicon (Si) is a trace element, which plays an important role in human bone growth. Si has been incorporated into biomaterials for bone regeneration in order to improve their osteogenic potential, both in vitro and in vivo. Little is known, however, as to how Si ions elicit their biological response on bone-forming cells. The aim of this study was to investigate the effect of Si ions on the proliferation, differentiation, bone-related gene expression and cell signalling pathways of bone marrow stromal cells (BMSCs) by comparing the BMSC responses to different concentrations of NaCl and Na2SiO3, while taking into account and excluding the effect of Na ions. Our study showed that Si ions at a concentration of 0.625 mM significantly enhanced the proliferation, mineralization nodule formation, bone-related gene expression (OCN, OPN and ALP) and bone matrix proteins (ALP and OPN) of BMSCs. Furthermore, Si ions at 0.625 mM could counteract the effect of the WNT inhibitor (W.I.) cardamonin on the osteogenic genes expression, (OPN, OCN and ALP), WNT and SHH signalling pathway-related genes in BMSCs. These results suggest that Si ions by themselves play an important role in regulating the proliferation and osteogenic differentiation of BMSCs, with the involvement of WNT and SHH signalling pathways. Our study provides evidence to explain possible molecular mechanisms whereby Si ions released from Si-containing biomaterials can acquire enhanced bioactivity at desired concentration.

Strategies for the prevention of cervical cancer by human papillomavirus vaccination

As cervical cancer is causally associated with 14 high-risk types of human papillomavirus (HPV), a successful HPV vaccine will have a major impact on this disease. Although some persistent HPV infections progress to cervical cancer, host immunity is generally able to clear most HPV infections. Both cell-mediated and antibody responses have been implicated in influencing the susceptibility, persistence or clearance of genital HPV infection. There have been two clinical trials that show that vaccines based on virus-like particles (VLPs) made from the major capsid protein, L1, are able to type specifically protect against cervical intra-epithelial neoplasia and infection. However, there is no evidence that even a mixed VLP vaccine will protect against types not included in the vaccine, and a major challenge that remains is how to engineer protection across a broader spectrum of viruses. Strategies for production of HPV vaccines using different vaccine vectors and different production systems are also reviewed. © 2005 Elsevier Ltd. All rights reserved.

Vaccination strategies for the prevention of cervical cancer

Infection with high-risk human papillomaviruses (HPVs) is an essential step in the multistep process leading to cervical cancer. There are approximately 120 different types of HPV identified: of these, 18 are high-risk types associated with cervical cancer, with HPV-16 being the dominant type in most parts of the world. The major capsid protein of papillomavirus, produced in a number of expression systems, self assembles to form virus-like particles. Virus-like particles are the basis of the first generation of HPV vaccines presently being tested in clinical trials. Virus-like particles are highly immunogenic and afford protection from infection both in animal models and in Phase IIb clinical trials. A number of Phase III trials are in progress to determine if the vaccine will protect against cervical disease and, in some cases, genital warts. However, it is predicted that these vaccines will be too expensive for the developing world, where they are desperately needed. Another problem is that they will be type specific. Novel approaches to the production of virus-like particles in plants, second-generation vaccine approaches including viral and bacterial vaccine vectors and DNA vaccines, as well as different routes of immunization, are also reviewed. © 2005 Future Drugs Ltd.

Preservice teachers’ observations of their mentors’ teaching strategies for differentiated learning

Tensions exist between teacher-centred and learner-centred approaches with constructivism as being favoured for learning in the 21st Century. There is little evidence of teaching strategies being used in the field for differentiating student learning. In addition, preservice teachers need to learn about teaching strategies for which observations of their mentor teachers can provide practical applications. This study explores 16 preservice teachers’ observations of their mentors’ teaching strategies over a four-week professional experience. They provided a minimum of five written observations during this period. Findings indicated that these preservice teachers observed their mentors’ practices and recorded four key teaching strategies used to differentiate learning, namely: (1) designating facilitators for students’ learning, including teacher, peers, parents, and support staff such as teachers aides, (2) managing student groups, (3) contexts for learning, and (4) using a range of teaching aids (visual, auditory, games) and resources. Preservice teachers’ observations of their mentor teachers indicated that they can commence at early stages for identifying teaching strategies and how they work for differentiating student learning.

Change-induced stressors and effective coping strategies in nursing

This study seeks to examine the causality of non-nursing and nursing stressors on the job satisfaction of nurses and how coping strategies have a mediating influence on this relationship in the context of sector-wide reform. To conceptualize the relationships, a mediation path model was developed. Data were collected at two time points using a self-completed online survey over a six monthly interval. During time 1, 306 Australian nurses completed the online survey. In the first wave (time 1), 306 Australian nurses completed the survey. In the second wave (time 2), matched survey data were collected from 119 nurses. The analysis showed a significant causal relationship between time 1 administrative and role stressors and an increase in nursing stress in time 2. A significant relationship was also identified between job specific context stressors and the adoption of effective coping strategies to deal with increased level of change-induced stress and strain and the likelihood of reporting higher level of job satisfaction in time 2. This study contributes by providing an integrated theoretical perspective on how stress affects retention that has so far been elusive. This is useful to researchers wanting to examine this phenomenon further and practitioners responsible for implementing change programs.

Scaffolds for growth factor delivery as applied to bone tissue engineering

There remains a substantial shortfall in treatment of severe skeletal injuries. The current gold standard of autologous bone grafting from the same patient, has many undesirable side effects associated such as donor site morbidity. Tissue engineering seeks to offer a solution to this problem. The primary requirements for tissue engineered scaffolds have already been well established, and many materials, such as polyesters, present themselves as potential candidates for bone defects; they have comparable structural features, but they often lack the required osteoconductivity to promote adequate bone regeneration. By combining these materials with biological growth factors; which promote the infiltration of cells into the scaffold as well as the differentiation into the specific cell and tissue type, it is possible to increase the formation of new bone. However cost and potential complications associated with growth factors means controlled release is an important consideration in the design of new bone tissue engineering strategies. This review will cover recent research in the area of encapsulation and release of growth factors within a variety of different polymeric scaffolds.