Optimization of human papillomavirus type 16 (HPV-16) L1 expression in plants: Comparison of the suitability of different HPV-16 L1 gene variants and different cell-compartment localization

Virus-like particle-based vaccines for high-risk human papillomaviruses (HPVs) appear to have great promise; however, cell culture-derived vaccines will probably be very expensive. The optimization of expression of different codon-optimized versions of the HPV-16 L1 capsid protein gene in plants has been explored by means of transient expression from a novel suite of Agrobacterium tumefaciens binary expression vectors, which allow targeting of recombinant protein to the cytoplasm, endoplasmic reticulum (ER) or chloroplasts. A gene resynthesized to reflect human codon usage expresses better than the native gene, which expresses better than a plant-optimized gene. Moreover, chloroplast localization allows significantly higher levels of accumulation of L1 protein than does cytoplasmic localization, whilst ER retention was least successful. High levels of L1 (>17% total soluble protein) could be produced via transient expression: the protein assembled into higher-order structures visible by electron microscopy, and a concentrated extract was highly immunogenic in mice after subcutaneous injection and elicited high-titre neutralizing antibodies. Transgenic tobacco plants expressing a human codon-optimized gene linked to a chloroplast-targeting signal expressed L1 at levels up to 11% of the total soluble protein. These are the highest levels of HPV L1 expression reported for plants: these results, and the excellent immunogenicity of the product, significantly improve the prospects of making a conventional HPV vaccine by this means. © 2007 SGM.

Expression of HIV-1 antigens in plants as potential subunit vaccines

Background Human immunodeficiency virus type 1 (HIV-1) has infected more than 40 million people worldwide, mainly in sub-Saharan Africa. The high prevalence of HIV-1 subtype C in southern Africa necessitates the development of cheap, effective vaccines. One means of production is the use of plants, for which a number of different techniques have been successfully developed. HIV-1 Pr55Gag is a promising HIV-1 vaccine candidate: we compared the expression of this and a truncated Gag (p17/p24) and the p24 capsid subunit in Nicotiana spp. using transgenic plants and transient expression via Agrobacterium tumefaciens and recombinant tobamovirus vectors. We also investigated the influence of subcellular localisation of recombinant protein to the chloroplast and the endoplasmic reticulum (ER) on protein yield. We partially purified a selected vaccine candidate and tested its stimulation of a humoral and cellular immune response in mice. Results Both transient and transgenic expression of the HIV antigens were successful, although expression of Pr55Gag was low in all systems; however, the Agrobacterium-mediated transient expression of p24 and p17/p24 yielded best, to more than 1 mg p24/kg fresh weight. Chloroplast targeted protein levels were highest in transient and transgenic expression of p24 and p17/p24. The transiently-expressed p17/p24 was not immunogenic in mice as a homologous vaccine, but it significantly boosted a humoral and T cell immune response primed by a gag DNA vaccine, pTHGagC. Conclusion Transient agroinfiltration was best for expression of all of the recombinant proteins tested, and p24 and p17/p24 were expressed at much higher levels than Pr55Gag. Our results highlight the usefulness of plastid signal peptides in enhancing the production of recombinant proteins meant for use as vaccines. The p17/p24 protein effectively boosted T cell and humoral responses in mice primed by the DNA vaccine pTHGagC, showing that this plant-produced protein has potential for use as a vaccine.

High level protein expression in plants through the use of a novel autonomously replicating geminivirus shuttle vector

We constructed a novel autonomously replicating gene expression shuttle vector, with the aim of developing a system for transiently expressing proteins at levels useful for commercial production of vaccines and other proteins in plants. The vector, pRIC, is based on the mild strain of the geminivirus Bean yellow dwarf virus (BeYDV-m) and is replicationally released into plant cells from a recombinant Agrobacterium tumefaciens Ti plasmid. pRIC differs from most other geminivirus-based vectors in that the BeYDV replication-associated elements were included in cis rather than from a co-transfected plasmid, while the BeYDV capsid protein (CP) and movement protein (MP) genes were replaced by an antigen encoding transgene expression cassette derived from the non-replicating A. tumefaciens vector, pTRAc. We tested vector efficacy in Nicotiana benthamiana by comparing transient cytoplasmic expression between pRIC and pTRAc constructs encoding either enhanced green fluorescent protein (EGFP) or the subunit vaccine antigens, human papillomavirus subtype 16 (HPV-16) major CP L1 and human immunodeficiency virus subtype C p24 antigen. The pRIC constructs were amplified in planta by up to two orders of magnitude by replication, while 50% more HPV-16 L1 and three- to seven-fold more EGFP and HIV-1 p24 were expressed from pRIC than from pTRAc. Vector replication was shown to be correlated with increased protein expression. We anticipate that this new high-yielding plant expression vector will contribute towards the development of a viable plant production platform for vaccine candidates and other pharmaceuticals. © 2009 Blackwell Publishing Ltd.

Human papillomavirus vaccines in plants

Human papillomaviruses are the etiological agents of cervical cancer, one of the two most prevalent cancers in women in developing countries. Currently available prophylactic vaccines are based on the L1 major capsid protein, which forms virus-like particles when expressed in yeast and insect cell lines. Despite their recognized efficacy, there are significant shortcomings: the vaccines are expensive, include only two oncogenic virus types, are delivered via intramuscular injection and require a cold chain. Plant expression systems may provide ways of overcoming some of these problems, in particular the expense. In this article, we report recent promising advances in the production of prophylactic and therapeutic vaccines against human papillomavirus by expression of the relevant antigens in plants, and discuss future prospects for the use of such vaccines. © 2010 Expert Reviews Ltd.

Riding the tide of biopharming in Africa: Considerations for risk assessment

In the past few years, plant biotechnology has gone beyond traditional agricultural production of food, feed and fibre, and moved to address more complex contemporary health, social and industrial challenges. The new era involves production of novel pharmaceutical products, speciality and fine chemicals, phytoremediation and production of renewable energy resources to replace non-renewable fossil fuels. Plants have been shown to provide a genuine and low-cost alternative production system for high-value products. Currently, the principal plant-made products include antibodies, feed additives, vaccine antigens and hormones for human and animal health, and industrial proteins. Despite the unique advantages of scalability, cost and product safety, issues of politics, environmental impact, regulation and socioeconomics still limit the adoption of biopharmaceuticals, especially in the developing world. Plant-based production systems have further complicated biosafety, gene flow and environmental impact assessments with generally genetically modified plants, topics that are already partially understood. This article provides a background to biopharming, highlighting basic considerations for risk assessment and regulation in developing countries, with an emphasis on plant-based vaccine production in South Africa.

Food insecurity among adults residing in disadvantaged urban areas : potential health and dietary consequences

Objective: Food insecurity is the limited or uncertain availability or access to nutritionally-adequate, culturally-appropriate and safe foods. Food insecurity may result in inadequate dietary intakes, overweight or obesity and the development of chronic disease. Internationally, few studies have focused on the range of potential health outcomes related to food insecurity among adults residing in disadvantaged locations and no such Australian studies exist. The objective of this study was to investigate associations between food insecurity, socio-demographic and health factors and dietary intakes among adults residing in disadvantaged urban areas. Design: Data were collected by mail survey (n= 505, 53% response rate), which ascertained information about food security status, demographic characteristics (such as age, gender, household income, education) fruit and vegetable intakes, take-away and meat consumption, general health, depression and chronic disease. Setting: Disadvantaged suburbs of Brisbane city, Australia, 2009. Subjects: Individuals aged ≥ 20 years. Results: Approximately one-in-four households (25%) were food insecure. Food insecurity was associated with lower household income, poorer general health, increased healthcare utilisation and depression. These associations remained after adjustment for age, gender and household income. Conclusion: Food insecurity is prevalent in urbanised disadvantaged areas in developed countries such as Australia. Low-income households are at high risk of experiencing food insecurity. Food insecurity may result in significant health burdens among the population, and this may be concentrated in socioeconomically-disadvantaged suburbs.

Associations between socioeconomic position, food insecurity and BMI

Introduction: Food insecurity is a social determinant of health and is defined as limited ability to access sufficient amounts of nutritionally adequate or safe food for a healthy and active life. Food insecurity is associated with poor health status and the exacerbation of other health inequalities. This study examined whether an association existed between 1) socioeconomic position (SEP) and food insecurity and 2) food insecurity and weight status. Methods: Data from the 1995 National Nutrition Survey was analysed. A random sample of households (n = 13 858) were asked about dietary habits and food choices. Information about gender, age, BMI, waist circumference, household income and whether the household had run out of money to purchase food in the previous 12 months was obtained and analysed using chi-square and logistic regression. Results: Income was significantly associated with food insecurity; households with lower income were at higher risk of food insecurity. Lower income males were nine times more likely to experience food insecurity and lower income females were three times more likely to experience food insecurity than their higher income counterparts. Food insecurity was significantly associated with body mass index (BMI) among women but not men. Women experiencing food insecurity were at higher risk of overweight/obesity according to BMI and waist circumference measures. Conclusion: Evidence suggests that low income households are at higher risk of food insecurity and women who are food insecure are at higher risk of being overweight or obese. Food insecurity may mediate the association between SEP and BMI.

Does food insecurity contribute to socioeconomic inequalities in BMI?

Introduction: Food insecurity is a social determinant of health and is defined as limited ability to access sufficient amounts of nutritionally adequate or safe food for a healthy and active life. Food insecurity is associated with poor health status and the exacerbation of other health inequalities. This study examined whether an association existed between 1) socioeconomic position (SEP) and food insecurity and 2) food insecurity and weight status. Methods: Data from the 1995 National Nutrition Survey was analysed. A random sample of households (n = 13 858) were asked about dietary habits and food choices. Information about gender, age, BMI, waist circumference, household income and whether the household had run out of money to purchase food in the previous 12 months was obtained and analysed using chi-square and logistic regression. Results: Income was significantly associated with food insecurity; households with lower income were at higher risk of food insecurity. Lower income males were nine times more likely to experience food insecurity and lower income females were three times more likely to experience food insecurity than their higher income counterparts. Food insecurity was significantly associated with body mass index (BMI) among women but not men. Women experiencing food insecurity were at higher risk of overweight/obesity according to BMI and waist circumference measures. Conclusion: Evidence suggests that low income households are at higher risk of food insecurity and women who are food insecure are at higher risk of being overweight or obese. Food insecurity may mediate the association between SEP and BMI.

Potential dietary consquences of food insecurity among Australian adults residing in urbanised disadvantaged areas

Introduction: Food insecurity is the limited/uncertain availability, access to or ability to acquire nutritionally-adequate, culturallyrelevant and safe foods. Adults suffering from food insecurity are at risk of inadequate nutrient intakes or, paradoxically, overweight/ obesity and the development of chronic disease. Despite the global financial crisis and rising costs of living, there are few studies investigating the potential dietary consequences of food insecurity among the Australian population. This study examined whether food insecurity was associated with weight status and poorer intakes of fruits, vegetable and takeaway foods among adults residing in socioeconomically-disadvantaged urbanised areas. Methods: In this cross-sectional study, a random sample of residents (n=1000) were selected from the most disadvantaged suburbs of Brisbane city (response rate 51%). Data were collected by postal questionnaire which ascertained information on sociodemographic information, household food security status, height, weight, fruit and vegetable intakes and takeaway consumption. Data were analysed using chi-square and logistic regression. Results: The overall prevalence of food insecurity was 31%. Food insecurity was not associated with weight status among men or women. Associations between food security status and potential dietary consequences differed for men and women. Among women, food security was not associated with intakes of fruit, vegetable or takeaway consumption. Contrastingly, among men food security was associated with vegetable intakes and consumption of takeaway food: men reporting food insecurity had lower intakes of vegetables and were more likely to consume takeaway foods compared to those that were food secure. Conclusion: Food security is an important public health issue in Australia and has potential dietary consequences that may adversely affect the health of food-insecure groups, most notably men residing in food-insecure households.

Potential health and dietary consequences of food insecurity among Australian adults residing in urbanised disadvantaged areas

Purpose: Food insecurity is the limited/uncertain availability or ability to acquire nutritionally-adequate, culturally-relevant and safe foods. Adults suffering from food insecurity are at risk of inadequate nutrient intakes or, paradoxically, overweight/obesity and the development of chronic disease. Despite the global financial crisis and rising costs of living, few studies have investigated the potential dietary and health consequences of food insecurity among the Australian population. This study examined whether food insecurity was associated with health behaviours and dietary intakes among adults residing in socioeconomically-disadvantaged urbanised areas. Methods: In this cross-sectional study, a random sample of residents (n = 1000) were selected from the most disadvantaged suburbs of Brisbane city (response rate 51%). Data were collected by postal questionnaire which ascertained information on socio-demographic information, household food security, height, weight, frequency of healthcare utilisation, presence of chronic disease and intakes of fruit, vegetables and take-away. Data were analysed using logistic regression. Results/Findings: The prevalence of food insecurity was 25%. Those reporting food insecurity were two-to-three times more likely to have seen a general practitioner or been hospitalised within the previous 6 months. Furthermore, food insecurity was associated with a three-to-six-fold increase in the likelihood of experiencing depression. Food insecurity was associated with higher intakes of some take-away foods, however was not significantly associated with weight status or intakes of fruits or vegetables among this disadvantaged sample. Conclusion: Food insecurity has potential adverse health consequences that may result in significant health burdens among the population, and this may be concentrated in socioeconomically-disadvantaged suburbs.

Evaluation of implementation of a healthy food and drink supply strategy throughout the whole school environment in Queensland state schools, Australia

BACKGROUND/OBJECTIVES: This paper reports on the evaluation of the Smart Choices healthy food and drink supply strategy for Queensland schools (Smart Choices) implementation across the whole school environment in state government primary and secondary schools in Queensland, Australia. SUBJECTS/METHODS: Three concurrent surveys using different methods for each group of stakeholders that targeted all 1275 school Principals, all 1258 Parent and Citizens’ Associations (P&Cs) and a random sample of 526 tuckshop convenors throughout Queensland. Nine hundred and seventy-three Principals, 598 P&Cs and 513 tuckshop convenors participated with response rates of 78%, 48% and 98%, respectively. RESULTS: Nearly all Principals (97%), P&Cs (99%) and tuckshop convenors (97%) reported that their school tuckshop had implemented Smart Choices. The majority of Principals and P&Cs reported implementation, respectively, in: school breakfast programs (98 and 92%); vending machine stock (94 and 83%); vending machine advertising (85 and 84%); school events (87 and 88%); school sporting events (81 and 80%); sponsorship and advertising (93 and 84%); fundraising events (80 and 84%); and sporting clubs (73 and 75%). Implementation in curriculum activities, classroom rewards and class parties was reported, respectively, by 97%, 86% and 75% of Principals. Respondents also reported very high levels of understanding of Smart Choices and engagement of the school community. CONCLUSIONS: The results demonstrated that food supply interventions to promote nutrition across all domains of the school environment can be implemented successfully.

Measurement of structural changes to a food material during dehydration

Rates of dehydration/rehydration are important quality parameters for dried products. Theoretically, if there are no adverse effects on the integrity of the tissue structure, it should absorb water to the same moisture content of the initial product before drying.The purpose of this work is to semi-automate the process of detection of cell structure boundaries as a food is dehydrated and rehydrated. This will enable food materials researchers to quantify changes to material’s structure as these processes take place. Images of potato cells as they were dehydrated and rehydrated were taken using an electron microscope. Cell boundaries were detected using an image processing algorithm. Average cell area and perimeter at each stage of dehydration were calculated and plotted versus time. The results show that the algorithm can successfully identify cell boundaries.

Smoking status as a moderator of the association between food insecurity and decreased fruit and vegetable consumption

Food insecurity is the inadequate access to, or availability of, sufficient amounts of nutritious, culturally-appropriate and safe foods, or the inability to acquire such foods by socially acceptable means. Food insecurity has been shown to be associated with poor dietary intakes and poor health status. Recently, evidence has emerged suggesting increased rates of food insecurity among those with substance abuse problems, including those who smoke. This cross-sectional study investigates the potential moderating effect of smoking on the association between food insecurity and fruit and vegetable intakes among the Australian population, using regression analyses. Participants were adults 18 years and older participating in the 2004/05 National Health Survey (n = 19,500). Those from food insecure households were up to two-times more likely to report inadequate fruit and vegetable intakes compared to those who were food secure. Those who smoked were nearly six times more likely to report being food insecure, and up to three-times more likely to report inadequate fruit and vegetable intakes, compared to their non-smoking counterparts. Further analyses revealed a marked decline in the strength of the association between food insecurity and fruit consumption with the addition of smoking status into a regression model. These findings have important implications for the development of policy and interventions to address food insecurity, suggesting that those from food insecure households are less likely to comply with national dietary recommendations, and that this may in part be moderated by smoking status.

Associations exist between food insecurity, poor diet, chronic illness and obesity among an Australian population

Food insecurity is the limited availability of, or access to, sufficient amounts of nutritious, culturally-appropriate and safe foods, or the inability to access such foods by socially-acceptable means. Evidence from the United States and Canada suggests that food insecurity may be associated with poor dietary intakes, obesity, and chronic disease including depression and diabetes, thus constituting a significant public health issue. Currently, no existing studies have investigated the dietary and health factors associated with food insecurity among the general Australian population. The current study investigated the potential associations between food insecurity, diet and health among Australian adults (18 years and over) participating in the cross-sectional National Health Survey (n = 19,500). Data were analysed by logistic regression adjusting for sociodemographic covariates. Those from food insecure households were 50% less likely to consume the recommended number of servings of fruit, 60% more likely to report poor health and experienced a 6-fold increase in the risk of severe depressive disorders, compared to their food secure counterparts. Furthermore, food insecurity was associated with a 50% increase in the risk of being diagnosed with ‘high sugar levels’. Finally, women from food insecure households were 30% more likely to be obese compared to their food secure counterparts. These findings suggest that food insecurity may play an important role in preventing adherence with national dietary recommendations, and may increase the risk of obesity and chronic illness. This has important implications for both clinical practice, and the development of interventions and policy to address food insecurity.