Optimization of chimeric HIV-1 virus-like particle production in a baculovirus-insect cell expression system

A baculovirus-insect cell expression system potentially provides the means to produce prophylactic HIV-1 virus-like particle (VLP) vaccines inexpensively and in large quantities. However, the system must be optimized to maximize yields and increase process efficiency. In this study, we optimized the production of two novel, chimeric HIV-1 VLP vaccine candidates (GagRT and GagTN) in insect cells. This was done by monitoring the effects of four specific factors on VLP expression: these were insect cell line, cell density, multiplicity of infection (MOI), and infection time. The use of western blots, Gag p24 ELISA, and four-factorial ANOVA allowed the determination of the most favorable conditions for chimeric VLP production, as well as which factors affected VLP expression most significantly. Both VLP vaccine candidates favored similar optimal conditions, demonstrating higher yields of VLPs when produced in the Trichoplusia ni Pro insect cell line, at a cell density of 1 × 106 cells/mL, and an infection time of 96 h post infection. It was found that cell density and infection time were major influencing factors, but that MOI did not affect VLP expression significantly. This work provides a potentially valuable guideline for HIV-1 protein vaccine optimization, as well as for general optimization of a baculovirus-based expression system to produce complex recombinant proteins. © 2009 American Institute of Chemical Engineers.

Spatial and temporal analysis of Barmah Forest virus disease in Queensland, Australia

Barmah Forest virus (BFV) disease is one of the most widespread mosquito-borne diseases in Australia. The number of outbreaks and the incidence rate of BFV in Australia have attracted growing concerns about the spatio-temporal complexity and underlying risk factors of BFV disease. A large number of notifications has been recorded continuously in Queensland since 1992. Yet, little is known about the spatial and temporal characteristics of the disease. I aim to use notification data to better understand the effects of climatic, demographic, socio-economic and ecological risk factors on the spatial epidemiology of BFV disease transmission, develop predictive risk models and forecast future disease risks under climate change scenarios. Computerised data files of daily notifications of BFV disease and climatic variables in Queensland during 1992-2008 were obtained from Queensland Health and Australian Bureau of Meteorology, respectively. Projections on climate data for years 2025, 2050 and 2100 were obtained from Council of Scientific Industrial Research Organisation. Data on socio-economic, demographic and ecological factors were also obtained from relevant government departments as follows: 1) socio-economic and demographic data from Australian Bureau of Statistics; 2) wetlands data from Department of Environment and Resource Management and 3) tidal readings from Queensland Department of Transport and Main roads. Disease notifications were geocoded and spatial and temporal patterns of disease were investigated using geostatistics. Visualisation of BFV disease incidence rates through mapping reveals the presence of substantial spatio-temporal variation at statistical local areas (SLA) over time. Results reveal high incidence rates of BFV disease along coastal areas compared to the whole area of Queensland. A Mantel-Haenszel Chi-square analysis for trend reveals a statistically significant relationship between BFV disease incidence rates and age groups (ƒÓ2 = 7587, p<0.01). Semi-variogram analysis and smoothed maps created from interpolation techniques indicate that the pattern of spatial autocorrelation was not homogeneous across the state. A cluster analysis was used to detect the hot spots/clusters of BFV disease at a SLA level. Most likely spatial and space-time clusters are detected at the same locations across coastal Queensland (p<0.05). The study demonstrates heterogeneity of disease risk at a SLA level and reveals the spatial and temporal clustering of BFV disease in Queensland. Discriminant analysis was employed to establish a link between wetland classes, climate zones and BFV disease. This is because the importance of wetlands in the transmission of BFV disease remains unclear. The multivariable discriminant modelling analyses demonstrate that wetland types of saline 1, riverine and saline tidal influence were the most significant risk factors for BFV disease in all climate and buffer zones, while lacustrine, palustrine, estuarine and saline 2 and saline 3 wetlands were less important. The model accuracies were 76%, 98% and 100% for BFV risk in subtropical, tropical and temperate climate zones, respectively. This study demonstrates that BFV disease risk varied with wetland class and climate zone. The study suggests that wetlands may act as potential breeding habitats for BFV vectors. Multivariable spatial regression models were applied to assess the impact of spatial climatic, socio-economic and tidal factors on the BFV disease in Queensland. Spatial regression models were developed to account for spatial effects. Spatial regression models generated superior estimates over a traditional regression model. In the spatial regression models, BFV disease incidence shows an inverse relationship with minimum temperature, low tide and distance to coast, and positive relationship with rainfall in coastal areas whereas in whole Queensland the disease shows an inverse relationship with minimum temperature and high tide and positive relationship with rainfall. This study determines the most significant spatial risk factors for BFV disease across Queensland. Empirical models were developed to forecast the future risk of BFV disease outbreaks in coastal Queensland using existing climatic, socio-economic and tidal conditions under climate change scenarios. Logistic regression models were developed using BFV disease outbreak data for the existing period (2000-2008). The most parsimonious model had high sensitivity, specificity and accuracy and this model was used to estimate and forecast BFV disease outbreaks for years 2025, 2050 and 2100 under climate change scenarios for Australia. Important contributions arising from this research are that: (i) it is innovative to identify high-risk coastal areas by creating buffers based on grid-centroid and the use of fine-grained spatial units, i.e., mesh blocks; (ii) a spatial regression method was used to account for spatial dependence and heterogeneity of data in the study area; (iii) it determined a range of potential spatial risk factors for BFV disease; and (iv) it predicted the future risk of BFV disease outbreaks under climate change scenarios in Queensland, Australia. In conclusion, the thesis demonstrates that the distribution of BFV disease exhibits a distinct spatial and temporal variation. Such variation is influenced by a range of spatial risk factors including climatic, demographic, socio-economic, ecological and tidal variables. The thesis demonstrates that spatial regression method can be applied to better understand the transmission dynamics of BFV disease and its risk factors. The research findings show that disease notification data can be integrated with multi-factorial risk factor data to develop build-up models and forecast future potential disease risks under climate change scenarios. This thesis may have implications in BFV disease control and prevention programs in Queensland.

What are the implications of peri-urban agriculture on food security in Australian cities?

Background: Periurban agriculture refers to agricultural practice occurring in areas with mixed rural and urban features. It is responsible 25% of the total gross value of economic production in Australia, despite only comprising 3% of the land used for agriculture. As populations grows and cities expand, they are constantly absorbing surrounding fringe areas, thus creating a new fringe, further from the city causing the periurban region to constantly shift outwards. Periurban regions are fundamental in the provision of fresh food to city populations and residential (and industrial) expansion taking over agricultural land has been noted as a major worldwide concern. Another major concern around the increase in urbanisation and resultant decrease in periurban agriculture is its potential effect on food security. Food security is the availability or access to nutritionally-adequate, culturally-relevant and safe foods in culturally-appropriate ways. Thus food insecurity occurs when access to or availability of these foods is compromised. There is an important level of connectedness between food security and food production and a decrease in periurban agriculture may have adverse effects on food security. A decrease in local, seasonal produce may result in a decrease in the availability of products and an increase in cost, as food must travel greater distances, incurring extra costs present at the consumer level. Currently, few Australian studies exist examining the change in periurban agriculture over time. Such information may prove useful for future health policy and interventions as well as infrastructure planning. The aim of this study is to investigate changes in periurban agriculture among capital cities of Australia. Methods: We compared data pertaining to selected commodities from the Australian Bureau of Statistics 2000-01 and 2005 -2006 Agricultural Census. This survey is distributed online or via mail on a five-yearly basis to approximately 175,000 Agricultural business to ascertain information on a range of factors, such as types of crops, livestock and land preparation practices. For the purpose of this study we compared the land being used for total crops, and cereal , oil seed, legume, fruit and vegetable crops separately. Data was analysed using repeated measures anova in spss. Results: Overall, total area available for crops in urbanised areas of Australia increased slightly by 1.8%. However, Sydney, Melbourne, Adelaide and Perth experienced decreases in the area available for fruit crops by 11%, 5%,and 4% respectively. Furthermore, Brisbane and Perth experienced decreases in land available for vegetable crops by 28% and 14% respectively. Finally, Sydney, Adelaide and Perth experienced decreases in land available for cereal crops by 10 – 79%. Conclusions: These findings suggest that population increases and consequent urban sprawl may be resulting in a decrease in peri-urban agriculture, specifically for several core food groups including fruit, breads and grain based foods. In doing so, access to or availability of these foods may be limited, and the cost of these foods is likely to increase, which may compromise food insecurity for certain sub-groups of the population.

Rolling in the aisles : a comparative study of male and female grocery shopper typologies

The purpose of this paper is to segment male and female grocery shoppers based on store and product attribute evaluations. A rich profile for each segment is developed. Gender comparisons are operationalised and these developed contemporary shopper typologies are further contrasted against earlier works. Data of 560 grocery shoppers was attained by a survey questionnaire. Factor analysis, cluster analysis and ANOVA were employed to develop specific segments of shoppers. Four distinct cohorts of male shoppers and three cohorts of female shoppers emerge from the data of eight constructs, measured by 46 items. One new shopper type, not found in earlier typology literature, emerged from this research. This shopper presented as a young, well educated, at the commencement of their career and family lifecycle, attracted by a strong value offer and willingness to share the family food shopping responsibilities. This research makes a contribution to segmentation literature and grocery retail practice in several ways. It presents the first retail typology of male supermarket shoppers, employing a cluster analysis technique. Comparisons between male and female grocery shopping typologies are accordingly facilitated. The research provides insights into the modern family food shopping behaviour of men; a channel in which men are now recognised as equal contributors. Research outcomes encourage supermarket retailers to implement targeted marketing and rationalized operational strategies that deliver on attributes of importance. Finally, this research provides the basis for further cross-cultural, cross-contextual comparative studies.

A coupled SPH-DEM model for fluid and solid mechanics of apple parenchyma cells during drying

A coupled SPH-DEM based two-dimensional (2-D) micro-scale single cell model is developed to predict basic cell-level shrinkage effects of apple parenchyma cells during air drying. In this newly developed drying model, Smoothed Particle Hydrodynamics (SPH) is used to model the low Reynolds Number fluid motions of the cell protoplasm, and a Discrete Element Method (DEM) is employed to simulate the polymer-like cell wall. Simulations results reasonably agree with published experimental drying results on cellular shrinkage properties such as cellular area, diameter and perimeter. These preliminary results indicate that the model is effective for the modelling and simulation of apple parenchyma cells during air drying.

A particle-based micromechanics approach to simulate structural changes of plant cells during drying

This paper is concerned with applying a particle-based approach to simulate the micro-level cellular structural changes of plant cells during drying. The objective of the investigation was to relate the micro-level structural properties such as cell area, diameter and perimeter to the change of moisture content of the cell. Model assumes a simplified cell which consists of two basic components, cell wall and cell fluid. The cell fluid is assumed to be a Newtonian fluid with higher viscosity compared to water and cell wall is assumed to be a visco-elastic solid boundary located around the cell fluid. Cell fluid is modelled with Smoothed Particle Hydrodynamics (SPH) technique and for the cell wall; a Discrete Element Method (DEM) is used. The developed model is two-dimensional, but accounts for three-dimensional physical properties of real plant cells. Drying phenomena is simulated as fluid mass reductions and the model is used to predict the above mentioned structural properties as a function of cell fluid mass. Model predictions are found to be in fairly good agreement with experimental data in literature and the particle-based approach is demonstrated to be suitable for numerical studies of drying related structural deformations. Also a sensitivity analysis is included to demonstrate the influence of key model parameters to model predictions.

Numerical simulation of red blood cells' deformation using SPH method

A numerical simulation method for the Red Blood Cells’ (RBC) deformation is presented in this study. The two-dimensional RBC membrane is modeled by the spring network, where the elastic stretch/compression energy and the bending energy are considered with the constraint of constant RBC surface area. Smoothed Particle Hydrodynamics (SPH) method is used to solve the Navier-Stokes equation coupled with the Plasma-RBC membrane and Cytoplasm- RBC membrane interaction. To verify the method, the motion of a single RBC is simulated in Poiseuille flow and compared with the results reported earlier. Typical motion and deformation mechanism of the RBC is observed.

A coupled SPH/FEM analysis of portable water filled barriers

Portable water filled barriers (PWFB) are semi-rigid roadside barriers which have the potential to display good crash attenuation characteristics at low and moderate impact speeds. The traditional mesh based numerical methods alone fail to simulate this type of impact with precision, stability and efficiency. This paper proposes to develop an advanced simulation model based on the combination of Smoothed Particles Hydrodynamics (SPH), a meshless method, and finite element method (FEM) for fluid-structure analysis using the commercially available software package LS-Dyna. The interaction between SPH particles and FEA elements is studied in this paper. Two methods of element setup at the element boundary were investigated. The response of the impacted barrier and fluid inside were analysed and compared. The system response and lagging were observed and reported in this paper. It was demonstrated that coupled SPH/FEM can be used in full scale PWFB modelling application. This will aid the research in determining the best initial setup to couple FEA and SPH in road safety barrier for impact response and safety analysis in the future.

Power of QTL detection using association tests with family controls

The power of testing for a population-wide association between a biallelic quantitative trait locus and a linked biallelic marker locus is predicted both empirically and deterministically for several tests. The tests were based on the analysis of variance (ANOVA) and on a number of transmission disequilibrium tests (TDT). Deterministic power predictions made use of family information, and were functions of population parameters including linkage disequilibrium, allele frequencies, and recombination rate. Deterministic power predictions were very close to the empirical power from simulations in all scenarios considered in this study. The different TDTs had very similar power, intermediate between one-way and nested ANOVAs. One-way ANOVA was the only test that was not robust against spurious disequilibrium. Our general framework for predicting power deterministically can be used to predict power in other association tests. Deterministic power calculations are a powerful tool for researchers to plan and evaluate experiments and obviate the need for elaborate simulation studies.

Caring efficacy and job satisfaction in Australian registered nurses

Purpose: The purpose of the study was to examine relationships between socio-demographic variables, job satisfaction and nurses’ caring experiences in a registered nurse population, as measured by the caring efficacy scale (CES) which was developed from Bandura’s social cognitive theory and Watson’s transpersonal caring theory. Methods: A cross-sectional survey was undertaken of nurses representing a variety of nursing specialties. A stratified random sample of registered nurses, who were members of a professional nursing organisation, was invited to participate in this study. Descriptive analyses, correlation analyses, one- way ANOVA tests, simple linear regression and multivariable analyses were conducted to examine if any relationships existed between these variables. Results: There were a total of 639 respondents to the national survey. The respondents (100%) showed positive perceived CES scores and 80.8% showed positive job satisfaction scores. Correlation analysis found age, years experience as a registered nurse and years in current job, all positively correlated with each other, (r >0.40: p < 0.001). CES scores were found to be positively correlated with age, years of experience as a registered nurse (r>0.1: p < 0.001) and job satisfaction (r>0.1: p < 0.001). An ANOVA found significant positive relationships between CES scores and age (p=0.05). Conclusion: Results from this study have identified that relationships between age, years of experience, job satisfaction and the perceived caring experiences of nurses’ exist. Organisational leaders may develop strategies for professional development and orientation programmes that enhance the caring experiences of nurses to provide quality patient care. The development of programmes that provide role modelling, emotional support or use verbal persuasion are needed where encouragement is required for nurses to master new skills. This may also improve job satisfaction and retention of nurses in the workplace in the current economically focussed healthcare system.

Examining the effects of an eco-driving system on driver distraction

Driver distraction has recently been defined by Regan as "the diversion of attention away from activities critical for safe driving toward a competing activity, which may result in insufficient or no attention to activities critical for safe driving (Regan, Hallett & Gordon, 2011, p.1780)". One source of distraction is in-vehicle devices, even though they might provide other benefits, e.g. navigation systems. Currently, eco-driving systems have been growing rapidly in popularity. These systems send messages to drivers so that driving performance can be improved in terms of fuel efficiency. However, there remain unanswered questions about whether eco-driving systems endanger drivers by distracting them. In this research, the CARRS-Q advanced driving simulator was used in order to provide safety for participants and meanwhile simulate real world driving. The distraction effects of tasks involving three different in-vehicle systems were investigated: changing a CD, entering a five digit number as a part of navigation task and responding to an eco-driving task. Driving in these scenarios was compared with driving in the absence of these distractions, and while drivers engaged in critical manoeuvres. In order to account for practice effects, the same scenarios were duplicated on a second day. The three in-vehicle systems were not the exact facsimiles of any particular existing system, but were designed to have similar characteristics to those of system available. In general, the results show that drivers’ mental workloads are significantly higher in navigation and CD changing scenarios in comparison to the two other scenarios, which implies that these two tasks impose more visual/manual and cognitive demands. However, eco-driving mental workload is still high enough to be called marginally significant (p ~ .05) across manoeuvres. Similarly, event detection tasks show that drivers miss significantly more events in the navigation and CD changing scenarios in comparison to both the baseline and eco-driving scenario across manoeuvres. Analysis of the practice effect shows that drivers’ baseline scenario and navigation scenario exhibit significantly less demand on the second day. However, the number of missed events across manoeuvres confirmed that drivers can detect significantly more events on the second day for all scenarios. Distraction was also examined separately for five groups of manoeuvres (straight, lane changing, overtaking, braking for intersections and braking for roundabouts), in two locations for each condition. Repeated measures mixed ANOVA results show that reading an eco-driving message can potentially impair driving performance. When comparing the three in–vehicle distractions tested, attending to an eco-driving message is similar in effect to the CD changing task. The navigation task degraded driver performance much more than these other sources of distraction. In lane changing manoeuvres, drivers’ missed response counts degraded when they engaged in reading eco-driving messages at the first location. However, drivers’ event detection abilities deteriorated less at the second lane changing location. In baseline manoeuvres (driving straight), participants’ mean minimum speed degraded more in the CD changing scenario. Drivers’ lateral position shifted more in both CD changing and navigation tasks in comparison with both eco-driving and baseline scenarios, so they were more visually distracting. Participants were better at event detection in baseline manoeuvres in comparison with other manoeuvres. When approaching an intersection, the navigation task caused more events to be missed by participants, whereas eco-driving messages seemed to make drivers less distracted. The eco-driving message scenario was significantly less distracting than the navigation system scenario (fewer missed responses) when participants commenced braking for roundabouts. To sum up, in spite of the finding that two other in-vehicle tasks are more distracting than the eco-driving task, the results indicate that even reading a simple message while driving could potentially lead to missing an important event, especially when executing critical manoeuvres. This suggests that in-vehicle eco-driving systems have the potential to contribute to increased crash risk through distraction. However, there is some evidence of a practice effect which suggests that future research should focus on performance with habitual rather than novel tasks. It is recommended that eco-driving messages be delivered to drivers off-line when possible.

A leap into foodservice satisfaction in the private acute care setting

Monitoring foodservice satisfaction is a risk management strategy for malnutrition in the acute care sector, as low satisfaction may be associated with poor intake. This study aimed to investigate the relationship between age and foodservice satisfaction in the private acute care setting. Patient satisfaction was assessed using a validated tool, the Acute Care Hospital Foodservice Patient Satisfaction Questionnaire for data collected 2008–2010 (n = 779) at a private hospital, Brisbane. Age was grouped into three categories; <50 years, 51–70 years and >70 years. Fisher’s exact test assessed independence of categorical responses and age group; ANOVA or Kruskal–Wallis test was used for continuous variables. Dichotomised responses were analysed using logistic regression and odds ratios (95% confidence interval, p < 0.05). Overall foodservice satisfaction (5 point scale) was high (≥4 out of 5) and was independent of age group (p = 0.377). There was an increasing trend with age in mean satisfaction scores for individual dimensions of foodservice; food quality (p < 0.001), meal service quality (p < 0.001), staff service issues (p < 0.001) and physical environment (p < 0.001). A preference for being able to choose different sized meals (59.8% > 70 years vs 40.6% ≤50 years; p < 0.001) and response to ‘the foods are just the right temperature’ (55.3% >70 years vs 35.9% ≤50 years; p < 0.001) was dependent on age. For the food quality dimension, based on dichotomised responses (satisfied or not), the odds of satisfaction was higher for >70 years (OR = 5.0, 95% CI: 1.8–13.8; <50 years referent). These results suggest that dimensions of foodservice satisfaction are associated with age and can assist foodservices to meet varying generational expectations of clients.

New and young firms Entrepreneurship policy and the role of government – evidence from the Federation of Small Businesses survey

Purpose – The purpose of this paper is to investigate whether new and young firms are different from older firms. This analysis is undertaken to explore general characteristics, use of external resources and growth orientation. Design/methodology/approach – Data from the 2008 UK Federation of Small Businesses survey provided 8,000 responses. Quantitative analysis identified significantly different characteristics of firms from 0-4, 4-9, 9-19 and 20+ years. Factor analysis was utilised to identify the advice sets, finance and public procurement customers of greatest interest, with ANOVA used to statistically compare firms in the identified age groups with different growth aspirations. Findings – The findings reveal key differences between new, young and older firms in terms of characteristics including business sector, owner/manager age, education/business experience, legal status, intellectual property and trading performance. New and young firms were more able to access beneficial resources in terms of finance and advice from several sources. New and young firms were also able to more easily access government and external finance, as well as government advice, but less able to access public procurement. Research limitations/implications – New and young firms are utilising external networks to access several resources for development purposes, and this differs for older firms. This suggests that a more explicit age-differentiated focus is required for government policies aimed at supporting firm growth. Originality/value – The study provides important baseline data for future quantitative and qualitative studies focused on the impact of firm age and government policy.

The effect of testing protocol on immature bovine thoracic spine segment stiffness

Introduction. In vitro spine biomechanical testing has been central to many advances in understanding the physiology and pathology of the human spine. Owing to the difficulty in obtaining sufficient numbers of human samples to conduct these studies, animal spines have been accepted as a substitute model. However, it is difficult to compare results from different studies, as they use different preparation, testing and data collection methods. The aim of this study was to identify the effect of repeated cyclic loading on bovine spine segment stiffness. It also aimed to quantify the effect of multiple freeze-thaw sequences, as many tests would be difficult to complete in a single session [1-3]. Materials and Methods. Thoracic spines from 6-8 week old calves were used. Each spine was dissected and divided into motion segments including levels T4-T11 (n=28). These were divided into two equal groups. Each segment was potted in polymethylemethacrylate. An Instron Biaxial materials testing machine with a custom made jig was used for testing. The segments were tested in flexion/extension, lateral bending and axial rotation at 37 degrees C and 100% humidity, using moment control to a maximum plus/minus 1.75 Nm with a loading rate of 0.3 Nm per second. Group (A) were tested with continuous repeated cyclic loading for 500 cycles with data recorded at cycles 3, 5, 10, 25, 100, 200, 300, 400 and 500. Group (B) were tested with 10 load cycles after each of 5 freeze thaw sequences. Data was collected from the tenth load cycle after each sequence. Statistical analysis of the data was performed using paired samples t-tests, ANOVA and generalized estimating equations. Results. The data were confirmed as having a normal distribution. 1. There were significant reductions in mean stiffness in flexion/extension (-20%; P=0.001) and lateral bending (-17%; P=0.009) over the 500 load cycles. However, there was no statistically significant change in axial rotation (P=0.152) 2. There was no statistically significant difference between mean stiffness over the five freeze-thaw sequences in flexion/extension (p=0.879) and axial rotation (p=0.07). However, there was a significant reduction in stiffness in lateral bending (-26%; p=0.007) Conclusion. Biomechanical testing of immature bovine spine motion segments requires careful interpretation. The effect of the number of load cycles as well as the number of freeze-thaw cycles on the stiffness of the motion segments depends on the axis of main movement.

A gender comparison of food shopper typologies

This research segments male and female food shoppers based on store and product attribute evaluations, enjoyment, planning, and shopping responsibility. A rich profile for each segment is developed from a sample of 580 primary food shoppers. Gender comparisons are operationalized and these developed food shopper typologies are contrasted against earlier works. Factor analysis, cluster analysis, and ANOVA were employed to develop specific segments of shoppers. This is the first study to identify specific groups of male food shoppers and contrast these to traditional female shoppers and earlier typographical works. The research provides the basis for further cross-cultural, cross-contextual comparative studies.