Application of an appearance-based intervention to improve sun protection outcomes of outdoor workers in Queensland, Australia

Outdoor workers are exposed to high levels of ultraviolet radiation (UVR) and may thus be at greater risk to experience UVR-related health effects such as skin cancer, sun burn, and cataracts. A number of intervention trials (n=14) have aimed to improve outdoor workers’ work-related sun protection cognitions and behaviours. Only one study however has reported the use of UV-photography as part of a multi-component intervention. This study was performed in the USA and showed long-term (12 months) improvements in work-related sun protection behaviours. Intervention effects of the other studies have varied greatly, depending on the population studied, intervention applied, and measurement of effect. Previous studies have not assessed whether: - Interventions are similarly effective for workers in stringent and less stringent policy organisations; - Policy effect is translated into workers’ leisure time protection; - Implemented interventions are effective in the long-term; - The facial UV-photograph technique is effective in Australian male outdoor workers without a large additional intervention package, and; - Such interventions will also affect workers’ leisure time sun-related cognitions and behaviours. Therefore, the present Protection of Outdoor Workers from Environmental Radiation [POWER]-study aimed to fill these gaps and had the objectives of: a) assessing outdoor workers’ sun-related cognitions and behaviours at work and during leisure time in stringent and less stringent sun protection policy environments; b) assessing the effect of an appearance-based intervention on workers’ risk perceptions, intentions and behaviours over time; c) assessing whether the intervention was equally effective within the two policy settings; and d) assessing the immediate post-intervention effect. Effectiveness was described in terms of changes in sun-related risk perceptions and intentions (as these factors were shown to be main precursors of behaviour change in many health promotion theories) and behaviour. The study purposefully selected and recruited two organisations with a large outdoor worker contingent in Queensland, Australia within a 40 kilometre radius of Brisbane. The two organisations differed in the stringency of implementation and reinforcement of their organisational sun protection policy. Data were collected from 154 male predominantly Australian born outdoor workers with an average age of 37 years and predominantly medium to fair skin (83%). Sun-related cognitions and behaviours of workers were assessed using self-report questionnaires at baseline and six to twelve months later. Variation in follow-up time was due to a time difference in the recruitment of the two organisations. Participants within each organisation were assigned to an intervention or control group. The intervention group participants received a one-off personalised Skin Cancer Risk Assessment Tool [SCRAT]-letter and a facial UV-photograph with detailed verbal information. This was followed by an immediate post-intervention questionnaire within three months of the start of the study. The control group only received the baseline and follow-up questionnaire. Data were analysed using a variety of techniques including: descriptive analyses, parametric and non-parametric tests, and generalised estimating equations. A 15% proportional difference observed was deemed of clinical significance, with the addition of reported statistical significance (p<0.05) where applicable. Objective 1: Assess and compare the current sun-related risk perceptions, intentions, behaviours, and policy awareness of outdoor workers in stringent and less stringent sun protection policy settings. Workers within the two organisations (stringent n=89 and less stringent n=65) were similar in their knowledge about skin cancer, self efficacy, attitudes, and social norms regarding sun protection at work and during leisure time. Participants were predominantly in favour of sun protection. Results highlighted that compared to workers in a less stringent policy organisation working for an organisation with stringent sun protection policies and practices resulted in more desirable sun protection intentions (less willing to tan p=0.03) ; actual behaviours at work (sufficient use of upper and lower body protection, headgear, and sunglasses (p<0.001 for all comparisons), and greater policy awareness (awareness of repercussions if Personal Protective Equipment (PPE) was not used, p<0.001)). However the effect of the work-related sun protection policy was found not to extend to leisure time sun protection. Objective 2: Compare changes in sun-related risk perceptions, intentions, and behaviours between the intervention and control group. The effect of the intervention was minimal and mainly resulted in a clinically significant reduction in work-related self-perceived risk of developing skin cancer in the intervention compared to the control group (16% and 32% for intervention and control group, respectively estimated their risk higher compared to other outdoor workers: , p=0.11). No other clinical significant effects were observed at 12 months follow-up. Objective 3: Assess whether the intervention was equally effective in the stringent sun protection policy organisation and the less stringent sun protection policy organisation. The appearance-based intervention resulted in a clinically significant improvement in the stringent policy intervention group participants’ intention to protect from the sun at work (workplace*time interaction, p=0.01). In addition to a reduction in their willingness to tan both at work (will tan at baseline: 17% and 61%, p=0.06, at follow-up: 54% and 33%, p=0.07, stringent and less stringent policy intervention group respectively. The workplace*time interaction was significant p<0.001) and during leisure time (will tan at baseline: 42% and 78%, p=0.01, at follow-up: 50% and 63%, p=0.43, stringent and less stringent policy intervention group respectively. The workplace*time interaction was significant p=0.01) over the course of the study compared to the less stringent policy intervention group. However, no changes in actual sun protection behaviours were found. Objective 4: Examine the effect of the intervention on level of alarm and concern regarding the health of the skin as well as sun protection behaviours in both organisations. The immediate post-intervention results showed that the stringent policy organisation participants indicated to be less alarmed (p=0.04) and concerned (p<0.01) about the health of their skin and less likely to show the facial UV-photograph to others (family p=0.03) compared to the less stringent policy participants. A clinically significantly larger proportion of participants from the stringent policy organisation reported they worried more about skin cancer (65%) and skin freckling (43%) compared to those in the less stringent policy organisation (46%,and 23% respectively , after seeing the UV-photograph). In summary the results of this study suggest that the having a stringent work-related sun protection policy was significantly related to for work-time sun protection practices, but did not extend to leisure time sun protection. This could reflect the insufficient level of sun protection found in the general Australian population during leisure time. Alternatively, reactance caused by being restricted in personal decisions through work-time policy could have contributed to lower leisure time sun protection. Finally, other factors could have also contributed to the less than optimal leisure time sun protection behaviours reported, such as unmeasured personal or cultural barriers. All these factors combined may have lead to reduced willingness to take proper preventive action during leisure time exposure. The intervention did not result in any measurable difference between the intervention and control groups in sun protection behaviours in this population, potentially due to the long lag time between the implementation of the intervention and assessment at 12-months follow-up. In addition, high levels of sun protection behaviours were found at baseline (ceiling effect) which left little room for improvement. Further, this study did not assess sunscreen use, which was the predominant behaviour assessed in previous effective appearance-based interventions trials. Additionally, previous trials were mainly conducted in female populations, whilst the POWER-study’s population was all male. The observed immediate post-intervention result could be due to more emphasis being placed on sun protection and risks related to sun exposure in the stringent policy organisation. Therefore participants from the stringent policy organisation could have been more aware of harmful effects of UVR and hence, by knowing that they usually protect adequately, not be as alarmed or concerned as the participants from the less stringent policy organisation. In conclusion, a facial UV-photograph and SCRAT-letter information alone may not achieve large changes in sun-related cognitions and behaviour, especially of assessed 6-12 months after the intervention was implemented and in workers who are already quite well protected. Differences found between workers in the present study appear to be more attributable to organisational policy. However, against a background of organisational policy, this intervention may be a useful addition to sun-related workplace health and safety programs. The study findings have been interpreted while respecting a number of limitations. These have included non-random allocation of participants due to pre-organised allocation of participants to study group in one organisation and difficulty in separating participants from either study group. Due to the transient nature of the outdoor worker population, only 105 of 154 workers available at baseline could be reached for follow-up. (attrition rate=32%). In addition the discrepancy in the time to follow-up assessment between the two organisations was a limitation of the current study. Given the caveats of this research, the following recommendations were made for future research: - Consensus should be reached to define "outdoor worker" in terms of time spent outside at work as well as in the way sun protection behaviours are measured and reported. - Future studies should implement and assess the value of the facial UV-photographs in a wide range of outdoor worker organisations and countries. - More timely and frequent follow-up assessments should be implemented in intervention studies to determine the intervention effect and to identify the best timing of booster sessions to optimise results. - Future research should continue to aim to target outdoor workers’ leisure time cognitions and behaviours and improve these if possible. Overall, policy appears to be an important factor in workers’ compliance with work-time use of sun protection. Given the evidence generated by this research, organisations employing outdoor workers should consider stringent implementation and reinforcement of a sun protection policy. Finally, more research is needed to improve ways to generate desirable behaviour in this population during leisure time.

Neuromuscular imbalances in the track sprinter

Athletic coaching can involve observation of a motor control task and then proposing guidance to an athlete about how the task performance can be developed. Coaches can identify the technique elements that seem to hinder performance and then provide instruction. Recently, a variety of training methods were proposed to enhance sprint performance, however a number of authors have identified these methods as characterised by low scientific evaluation or support (Brown & Vescovi, 2012; Jones, Bezodis, & Thompson, 2009). This article will outline a scientifically robust neuromuscular theory underlying poor movement techniques that may be visible when coaches observe sprint performance. The goal of this article is to inform the sprint coach of a method to identify and correct suboptimal biomechanics to enhance athletic performance.

Can you describe him for me? A technique for semantic person search in video

From a law enforcement standpoint, the ability to search for a person matching a semantic description (i.e. 1.8m tall, red shirt, jeans) is highly desirable. While a significant research effort has focused on person re-detection (the task of identifying a previously observed individual in surveillance video), these techniques require descriptors to be built from existing image or video observations. As such, person re-detection techniques are not suited to situations where footage of the person of interest is not readily available, such as a witness reporting a recent crime. In this paper, we present a novel framework that is able to search for a person based on a semantic description. The proposed approach uses size and colour cues, and does not require a person detection routine to locate people in the scene, improving utility in crowded conditions. The proposed approach is demonstrated with a new database that will be made available to the research community, and we show that the proposed technique is able to correctly localise a person in a video based on a simple semantic description.

Nanoparticles in European cities and associated health impacts

Atmospheric nanoparticles are one of those pollutants currently unregulated through ambient air quality standards. The aim of this chapter is to assess the environmental and health impacts of atmospheric nanoparticles in European environments. The chapter begins with the conventional information on the origin of atmospheric nanoparticles, followed by their physical and chemical characteristics. A brief overview of recently published review articles on this topic is then presented to guide those readers interested in exploring any specific aspect of nanoparticles in greater detail. A further section reports a summary of recently published studies on atmospheric nanoparticles in European cities. This covers a total of about 45 sampling locations in 30 different cities within 15 European countries for quantifying levels of roadside and urban background particle number concentrations (PNCs). Average PNCs at roadside and urban background sites were found to be 3.82±3.25 ×104 cm–3 and 1.63±0.82 ×104 cm–3, respectively, giving a roadside to background PNC ratio of ~2.4. Engineered nanoparticles are one of the key emerging categories of airborne nanoparticles, especially for the indoor environments. Their ambient concentrations may increase in future due to widespread use of nanotechnology integrated products. Evaluation of their sources and probable impacts on air quality and human health are briefly discussed in the following section. Respiratory deposition doses received by the public exposed to roadside PNCs in numerous European locations are then estimated. These were found to be in the 1.17–7.56 1010 h–1 range over the studied roadside European locations. The following section discusses the potential framework for airborne nanoparticle regulations in Europe and, in addition, the existing control measures to limit nanoparticle emissions at source. The chapter finally concludes with a synthesis of the topic areas covered and highlights important areas for further work.

Numerical solution of stress intensity factors of multiple cracks in great number with eigen COD boundary integral equations

Based on the eigen crack opening displacement (COD) boundary integral equations, a newly developed computational approach is proposed for the analysis of multiple crack problems. The eigen COD particularly refers to a crack in an infinite domain under fictitious traction acting on the crack surface. With the concept of eigen COD, the multiple cracks in great number can be solved by using the conventional displacement discontinuity boundary integral equations in an iterative fashion with a small size of system matrix. The interactions among cracks are dealt with by two parts according to the distances of cracks to the current crack. The strong effects of cracks in adjacent group are treated with the aid of the local Eshelby matrix derived from the traction BIEs in discrete form. While the relatively week effects of cracks in far-field group are treated in the iteration procedures. Numerical examples are provided for the stress intensity factors of multiple cracks, up to several thousands in number, with the proposed approach. By comparing with the analytical solutions in the literature as well as solutions of the dual boundary integral equations, the effectiveness and the efficiencies of the proposed approach are verified.

The relationship between Bayesian motor unit number estimation and histological measurements of motor neurons in wild-type and SOD1 G93A mice

Objective: To assess the relationship between Bayesian MUNE and histological motor neuron counts in wild-type mice and in an animal model of ALS. Methods: We performed Bayesian MUNE paired with histological counts of motor neurons in the lumbar spinal cord of wild-type mice and transgenic SOD1 G93A mice that show progressive weakness over time. We evaluated the number of acetylcholine endplates that were innervated by a presynaptic nerve. Results: In wild-type mice, the motor unit number in the gastrocnemius muscle estimated by Bayesian MUNE was approximately half the number of motor neurons in the region of the spinal cord that contains the cell bodies of the motor neurons supplying the hindlimb crural flexor muscles. In SOD1 G93A mice, motor neuron numbers declined over time. This was associated with motor endplate denervation at the end-stage of disease. Conclusion: The number of motor neurons in the spinal cord of wild-type mice is proportional to the number of motor units estimated by Bayesian MUNE. In SOD1 G93A mice, there is a lower number of estimated motor units compared to the number of spinal cord motor neurons at the end-stage of disease, and this is associated with disruption of the neuromuscular junction. Significance: Our finding that the Bayesian MUNE method gives estimates of motor unit numbers that are proportional to the numbers of motor neurons in the spinal cord supports the clinical use of Bayesian MUNE in monitoring motor unit loss in ALS patients. © 2012 International Federation of Clinical Neurophysiology.

Coordinated tuning of DFIG-based wind turbines and batteries using bacteria foraging technique for maintaining constant grid power output

This paper proposes the use of battery energy storage (BES) system for the grid-connected doubly fed induction generator (DFIG). The BES would help in storing/releasing additional power in case of higher/lower wind speed to maintain constant grid power. The DC link capacitor is replaced with the BES system in a DFIG-based wind turbine to achieve the above-mentioned goal. The control scheme is modified and the co-ordinated tuning of the associated controllers to enhance the damping of the oscillatory modes is presented using bacterial foraging technique. The results from eigenvalue analysis and the time domain simulation studies are presented to elucidate the effectiveness of the BES systems in maintaining the grid stability under normal operation.

Semi-parametric modelling of ultrafine particle number concentration

Quantifying spatial and/or temporal trends in environmental modelling data requires that measurements be taken at multiple sites. The number of sites and duration of measurement at each site must be balanced against costs of equipment and availability of trained staff. The split panel design comprises short measurement campaigns at multiple locations and continuous monitoring at reference sites [2]. Here we present a modelling approach for a spatio-temporal model of ultrafine particle number concentration (PNC) recorded according to a split panel design. The model describes the temporal trends and background levels at each site. The data were measured as part of the “Ultrafine Particles from Transport Emissions and Child Health” (UPTECH) project which aims to link air quality measurements, child health outcomes and a questionnaire on the child’s history and demographics. The UPTECH project involves measuring aerosol and particle counts and local meteorology at each of 25 primary schools for two weeks and at three long term monitoring stations, and health outcomes for a cohort of students at each school [3].

The influence of oxygenated organic aerosols (OOA) and its volatile organic content on the oxidative potential of diesel particulate mater

Airborne particulate matter pollution is of concern for a number of reasons and has been widely recognised as an important risk factor to human health. A number of toxicological and epidemiological studies reported negative health effects on both respiratory and cardiovascular system. Despite the availability of a huge body of research, the underlying toxicological mechanisms by which particles induce adverse health effects are not yet entirely understood. The production of reactive oxygen species (ROS) has been shown to induce oxidative stress, which is proposed as a mechanism for many of the adverse health outcomes associated with exposure to particulate matter (PM). Therefore, it is crucial to introduce a technique that will allow rapid and routine screenings of the oxidative potential of PM.

History behind the IVF egg: Are women's comprehensive histories associated with the number of eggs collected, or fertilised normally, from repeated IVF cycles? [abstract]

Most studies of in vitro fertilisation (IVF) outcomes use cycle-based data and fail to account for women who use repeated IVF cycles. The objective of this study was to examine the association between the number of eggs collected (EC) and the percentage fertilised normally, and women’s self-reported medical, personal and social histories. This study involved a crosssectional survey of infertile women (aged 27-46 years) recruited from four privately-owned fertility clinics located in major cities of Australia. Regression modeling was used to estimate the mean EC and mean percentage of eggs fertilised normally: adjusted for age at EC. Appropriate statistical methods were used to take account of repeated IVF cycles by the same women. Among 121 participants who returned the survey and completed 286 IVF cycles, the mean age at EC was 35.2 years (SD 4.5). Women’s age at EC was strongly associated with the number of EC: <30 years, 11.7 EC; 30.0-< 35 years, 10.6 EC; 35.0-<40.0 years, 7.3 EC; 40.0+ years, 8.1 EC; p<.0001. Prolonged use of oral contraceptives was associated with lower numbers of EC: never used, 14.6 EC; 0-2 years, 11.7 EC; 3-5 years, 8.5 EC; 6þ years, 8.2 EC; p=.04. Polycystic ovary syndrome (PCOS) was associated with more EC: have PCOS, 11.5 EC; no, 8.3 EC; p=.01. Occupational exposures may be detrimental to normal fertilisation: professional roles, 58.8%; trade and service roles, 51.8%; manual and other roles, 63.3%; p=.02. In conclusion, women’s age remains the most significant characteristic associated with EC but not the percentage of eggs fertilised normally.

Optimal image sample size for corneal nerve morphometry

Purpose Arbitrary numbers of corneal confocal microscopy images have been used for analysis of corneal subbasal nerve parameters under the implicit assumption that these are a representative sample of the central corneal nerve plexus. The purpose of this study is to present a technique for quantifying the number of random central corneal images required to achieve an acceptable level of accuracy in the measurement of corneal nerve fiber length and branch density. Methods Every possible combination of 2 to 16 images (where 16 was deemed the true mean) of the central corneal subbasal nerve plexus, not overlapping by more than 20%, were assessed for nerve fiber length and branch density in 20 subjects with type 2 diabetes and varying degrees of functional nerve deficit. Mean ratios were calculated to allow comparisons between and within subjects. Results In assessing nerve branch density, eight randomly chosen images not overlapping by more than 20% produced an average that was within 30% of the true mean 95% of the time. A similar sampling strategy of five images was 13% within the true mean 80% of the time for corneal nerve fiber length. Conclusions The “sample combination analysis” presented here can be used to determine the sample size required for a desired level of accuracy of quantification of corneal subbasal nerve parameters. This technique may have applications in other biological sampling studies.

Performance evaluation of onsite sewage treatment : results of soil investigations

Background The onsite treatment of sewage and effluent disposal is widely prevalent in rural and urban fringe areas due to the general unavailability of reticulated wastewater collection systems. Despite the low technology of the systems, failure is common and in many cases leading to adverse public health and environmental consequences. It is important therefore that careful consideration is given to the design and location of onsite sewage treatment systems. This requires an understanding of the factors that influence treatment performance. The use of subsurface absorption systems is the most common form of effluent disposal for onsite sewage treatment, particularly for septic tanks. Also, in the case of septic tanks, a subsurface disposal system is generally an integral component of the sewage treatment process. Site specific factors play a key role in the onsite treatment of sewage. The project The primary aims of the research project were: • to relate treatment performance of onsite sewage treatment systems to soil conditions at site; • to evaluate current research relating to onsite sewage treatment; and, • to identify key issues where currently there is a lack of relevant research. These tasks were undertaken with the objective of facilitating the development of performance based planning and management strategies for onsite sewage treatment. The primary focus of this research project has been on septic tanks. By implication, the investigation has been confined to subsurface soil absorption systems. The design and treatment processes taking place within the septic tank chamber itself did not form a part of the investigation. Five broad categories of soil types prevalent in the Brisbane region have been considered in this project. The number of systems investigated was based on the proportionate area of urban development within the Brisbane region located on each of the different soil types. In the initial phase of the investigation, the majority of the systems evaluated were septic tanks. However, a small number of aerobic wastewater treatment systems (AWTS) were also included. The primary aim was to compare the effluent quality of systems employing different generic treatment processes. It is important to note that the number of each different type of system investigated was relatively small. Consequently, this does not permit a statistical analysis to be undertaken of the results obtained for comparing different systems. This is an important issue considering the large number of soil physico-chemical parameters and landscape factors that can influence treatment performance and their wide variability. The report This report is the last in a series of three reports focussing on the performance evaluation of onsite treatment of sewage. The research project was initiated at the request of the Brisbane City Council. The project component discussed in the current report outlines the detailed soil investigations undertaken at a selected number of sites. In the initial field sampling, a number of soil chemical properties were assessed as indicators to investigate the extent of effluent flow and to help understand what soil factors renovate the applied effluent. The soil profile attributes, especially texture, structure and moisture regime were examined more in an engineering sense to determine the effect of movement of water into and through the soil. It is important to note that it is not only the physical characteristics, but also the chemical characteristics of the soil as well as landscape factors play a key role in the effluent renovation process. In order to understand the complex processes taking place in a subsurface effluent disposal area, influential parameters were identified using soil chemical concepts. Accordingly, the primary focus of this final phase of the research project was to identify linkages between various soil chemical parameters and landscape patterns and their contribution to the effluent renovation process. The research outcomes will contribute to the development of robust criteria for evaluating the performance of subsurface effluent disposal systems. The outcomes The key findings from the soil investigations undertaken are: • Effluent renovation is primarily undertaken by a combination of various soil physico-chemical parameters and landscape factors, thereby making the effluent renovation processes strongly site dependent. • Decisions regarding site suitability for effluent disposal should not be based purely in terms of the soil type. A number of other factors such as the site location in the catena, the drainage characteristics and other physical and chemical characteristics, also exert a strong influence on site suitability. • Sites, which are difficult to characterise in terms of suitability for effluent disposal, will require a detailed soil physical and chemical analysis to be undertaken to a minimum depth of at least 1.2 m. • The Ca:Mg ratio and Exchangeable Sodium Percentage are important parameters in soil suitability assessment. A Ca:Mg ratio of less than 0.5 would generally indicate a high ESP. This in turn would mean that Na and possibly Mg are the dominant exchangeable cations, leading to probable clay dispersion. • A Ca:Mg ratio greater than 0.5 would generally indicate a low ESP in the profile, which in turn indicates increased soil stability. • In higher clay percentage soils, low ESP can have a significant effect. • The presence of high exchangeable Na can be counteracted by the presence of swelling clays, and an exchange complex co-dominated by exchangeable Ca and exchangeable Mg. This aids absorption of cations at depth, thereby reducing the likelihood of dispersion. • Salt is continually added to the soil by the effluent and problems may arise if the added salts accumulate to a concentration that is harmful to the soil structure. Under such conditions, good drainage is essential in order to allow continuous movement of water and salt through the profile. Therefore, for a site to be sustainable, it would have a maximum application rate of effluent. This would be dependent on subsurface characteristics and the surface area available for effluent disposal. • The dosing regime for effluent disposal can play a significant role in the prevention of salt accumulation in the case of poorly draining sites. Though intermittent dosing was not considered satisfactory for the removal of the clogging mat layer, it has positive attributes in the context of removal of accumulated salts in the soil.

Spatial variation of particle number concentration in school microscale environment

There is significant toxicological evidence of the effects of ultrafine particles (<100nm) on human health (WHO 2005). Studies show that the number concentration of particles has been associated with adverse human health effects (Englert 2004). This work is part of a major study called ‘Ultrafine Particles form Traffic Emissions and Children’s Health’ (UPTECH), which seeks to determine the effect of the exposure to traffic related ultrafine particles on children’s health in schools (http://www.ilaqh.qut.edu.au/Misc/UPT ECH%20Home.htm). Quantification of spatial variation of particle number concentration (PNC) in a microscale environment and identification of the main affecting parameters and their contribution levels are the main aims of this analysis.

Revision total hip replacement using the cement-in-cement technique for the acetabular component : technique and results for 60 hips

The technique of femoral cement-in-cement revision is well established, but there are no previous series reporting its use on the acetabular side at the time of revision total hip arthroplasty. We describe the surgical technique and report the outcome of 60 consecutive cement-in-cement revisions of the acetabular component at a mean follow-up of 8.5 years (range 5-12 years). All had a radiologically and clinically well fixed acetabular cement mantle at the time of revision. 29 patients died. No case was lost to follow-up. The 2 most common indications for acetabular revision were recurrent dislocation (77%) and to compliment a femoral revision (20%). There were 2 cases of aseptic cup loosening (3.3%) requiring re-revision. No other hip was clinically or radiologically loose (96.7%) at latest follow-up. One case was re-revised for infection, 4 for recurrent dislocation and 1 for disarticulation of a constrained component. At 5 years, the Kaplan-Meier survival rate was 100% for aseptic loosening and 92.2% (95% CI; 84.8-99.6%) with revision for all causes as the endpoint. These results support the use of the cement-in-cement revision technique in appropriate cases on the acetabular side. Theoretical advantages include preservation of bone stock, reduced operating time, reduced risk of complications and durable fixation.

Optimisation of Banana streak virus (BSV) diagnostic assays

Bananas are one of the world�fs most important crops, serving as a staple food and an important source of income for millions of people in the subtropics. Pests and diseases are a major constraint to banana production. To prevent the spread of pests and disease, farmers are encouraged to use disease�] and insect�]free planting material obtained by micropropagation. This option, however, does not always exclude viruses and concern remains on the quality of planting material. Therefore, there is a demand for effective and reliable virus indexing procedures for tissue culture (TC) material. Reliable diagnostic tests are currently available for all of the economically important viruses of bananas with the exception of Banana streak viruses (BSV, Caulimoviridae, Badnavirus). Development of a reliable diagnostic test for BSV is complicated by the significant serological and genetic variation reported for BSV isolates, and the presence of endogenous BSV (eBSV). Current PCR�] and serological�]based diagnostic methods for BSV may not detect all species of BSV, and PCR�]based methods may give false positives because of the presence of eBSV. Rolling circle amplification (RCA) has been reported as a technique to detect BSV which can also discriminate between episomal and endogenous BSV sequences. However, the method is too expensive for large scale screening of samples in developing countries, and little information is available regarding its sensitivity. Therefore the development of reliable PCR�]based assays is still considered the most appropriate option for large scale screening of banana plants for BSV. This MSc project aimed to refine and optimise the protocols for BSV detection, with a particular focus on developing reliable PCR�]based diagnostics Initially, the appropriateness and reliability of PCR and RCA as diagnostic tests for BSV detection were assessed by testing 45 field samples of banana collected from nine districts in the Eastern region of Uganda in February 2010. This research was also aimed at investigating the diversity of BSV in eastern Uganda, identifying the BSV species present and characterising any new BSV species. Out of the 45 samples tested, 38 and 40 samples were considered positive by PCR and RCA, respectively. Six different species of BSV, namely Banana streak IM virus (BSIMV), Banana streak MY virus (BSMYV), Banana streak OL virus (BSOLV), Banana streak UA virus (BSUAV), Banana streak UL virus (BSULV), Banana streak UM virus (BSUMV), were detected by PCR and confirmed by RCA and sequencing. No new species were detected, but this was the first report of BSMYV in Uganda. Although RCA was demonstrated to be suitable for broad�]range detection of BSV, it proved time�]consuming and laborious for identification in field samples. Due to the disadvantages associated with RCA, attempts were made to develop a reliable PCR�]based assay for the specific detection of episomal BSOLV, Banana streak GF virus (BSGFV), BSMYV and BSIMV. For BSOLV and BSGFV, the integrated sequences exist in rearranged, repeated and partially inverted portions at their site of integration. Therefore, for these two viruses, primers sets were designed by mapping previously published sequences of their endogenous counterparts onto published sequences of the episomal genomes. For BSOLV, two primer sets were designed while, for BSGFV, a single primer set was designed. The episomalspecificity of these primer sets was assessed by testing 106 plant samples collected during surveys in Kenya and Uganda, and 33 leaf samples from a wide range of banana cultivars maintained in TC at the Maroochy Research Station of the Department of Employment, Economic Development and Innovation (DEEDI), Queensland. All of these samples had previously been tested for episomal BSV by RCA and for both BSOLV and BSGFV by PCR using published primer sets. The outcome from these analyses was that the newly designed primer sets for BSOLV and BSGFV were able to distinguish between episomal BSV and eBSV in most cultivars with some B�]genome component. In some samples, however, amplification was observed using the putative episomal�]specific primer sets where episomal BSV was not identified using RCA. This may reflect a difference in the sensitivity of PCR compared to RCA, or possibly the presence of an eBSV sequence of different conformation. Since the sequences of the respective eBSV for BSMYV and BSIMV in the M. balbisiana genome are not available, a series of random primer combinations were tested in an attempt to find potential episomal�]specific primer sets for BSMYV and BSIMV. Of an initial 20 primer combinations screened for BSMYV detection on a small number of control samples, 11 primers sets appeared to be episomal�]specific. However, subsequent testing of two of these primer combinations on a larger number of control samples resulted in some inconsistent results which will require further investigation. Testing of the 25 primer combinations for episomal�]specific detection of BSIMV on a number of control samples showed that none were able to discriminate between episomal and endogenous BSIMV. The final component of this research project was the development of an infectious clone of a BSV endemic in Australia, namely BSMYV. This was considered important to enable the generation of large amounts of diseased plant material needed for further research. A terminally redundant fragment (.1.3 �~ BSMYV genome) was cloned and transformed into Agrobacterium tumefaciens strain AGL1, and used to inoculate 12 healthy banana plants of the cultivars Cavendish (Williams) by three different methods. At 12 weeks post�]inoculation, (i) four of the five banana plants inoculated by corm injection showed characteristic BSV symptoms while the remaining plant was wilting/dying, (ii) three of the five banana plants inoculated by needle�]pricking of the stem showed BSV symptoms, one plant was symptomless while the remaining had died and (iii) both banana plants inoculated by leaf infiltration were symptomless. When banana leaf samples were tested for BSMYV by PCR and RCA, BSMYV was confirmed in all banana plants showing symptoms including those were wilting and/or dying. The results from this research have provided several avenues for further research. By completely sequencing all variants of eBSOLV and eBSGFV and fully sequencing the eBSIMV and eBSMYV regions, episomal BSV�]specific primer sets for all eBSVs could potentially be designed that could avoid all integrants of that particular BSV species. Furthermore, the development of an infectious BSV clone will enable large numbers of BSVinfected plants to be generated for the further testing of the sensitivity of RCA compared to other more established assays such as PCR. The development of infectious clones also opens the possibility for virus induced gene silencing studies in banana.