Investigation of highly regulated promoters for the production of recombinant proteins in plants

Plants have been identified as promising expression systems for the commercial production of recombinant proteins. Plant-based protein production or “biofarming” offers a number of advantages over traditional expression systems in terms of scale of production, the capacity for post-translation processing, providing a product free of contaminants and cost effectiveness. A number of pharmaceutically important and commercially valuable proteins, such as antibodies, biopharmaceuticals and industrial enzymes are currently being produced in plant expression systems. However, several challenges still remain to improve recombinant protein yield with no ill effect on the host plant. The ability for transgenic plants to produce foreign proteins at commercially viable levels can be directly related to the level and cell specificity of the selected promoter driving the transgene. The accumulation of recombinant proteins may be controlled by a tissue-specific, developmentally-regulated or chemically-inducible promoter such that expression of recombinant proteins can be spatially- or temporally- controlled. The strict control of gene expression is particularly useful for proteins that are considered toxic and whose expression is likely to have a detrimental effect on plant growth. To date, the most commonly used promoter in plant biotechnology is the cauliflower mosaic virus (CaMV) 35S promoter which is used to drive strong, constitutive transgene expression in most organs of transgenic plants. Of particular interest to researchers in the Centre for Tropical Crops and Biocommodities at QUT are tissue-specific promoters for the accumulation of foreign proteins in the roots, seeds and fruit of various plant species, including tobacco, banana and sugarcane. Therefore this Masters project aimed to isolate and characterise root- and seed-specific promoters for the control of genes encoding recombinant proteins in plant-based expression systems. Additionally, the effects of matching cognate terminators with their respective gene promoters were assessed. The Arabidopsis root promoters ARSK1 and EIR1 were selected from the literature based on their reported limited root expression profiles. Both promoters were analysed using the PlantCARE database to identify putative motifs or cis-acting elements that may be associated with this activity. A number of motifs were identified in the ARSK1 promoter region including, WUN (wound-inducible), MBS (MYB binding site), Skn-1, and a RY core element (seed-specific) and in the EIR1 promoter region including, Skn-1 (seed-specific), Box-W1 (fungal elicitor), Aux-RR core (auxin response) and ABRE (ABA response). However, no previously reported root-specific cis-acting elements were observed in either promoter region. To confirm root specificity, both promoters, and truncated versions, were fused to the GUS reporter gene and the expression cassette introduced into Arabidopsis via Agrobacterium-mediated transformation. Despite the reported tissue-specific nature of these promoters, both upstream regulatory regions directed constitutive GUS expression in all transgenic plants. Further, similar levels of GUS expression from the ARSK1 promoter were directed by the control CaMV 35S promoter. The truncated version of the EIR1 promoter (1.2 Kb) showed some differences in the level of GUS expression compared to the 2.2 Kb promoter. Therefore, this suggests an enhancer element is contained in the 2.2 Kb upstream region that increases transgene expression. The Arabidopsis seed-specific genes ATS1 and ATS3 were selected from the literature based on their seed-specific expression profiles and gene expression confirmed in this study as seed-specific by RT-PCR analysis. The selected promoter regions were analysed using the PlantCARE database in order to identify any putative cis elements. The seed-specific motifs GCN4 and Skn-1 were identified in both promoter regions that are associated with elevated expression levels in the endosperm. Additionaly, the seed-specific RY element and the ABRE were located in the ATS1 promoter. Both promoters were fused to the GUS reporter gene and used to transform Arabidopsis plants. GUS expression from the putative promoters was consitutive in all transgenic Arabidopsis tissue tested. Importantly, the positive control FAE1 seed-specific promoter also directed constitutive GUS expression throughout transgenic Arabidopsis plants. The constitutive nature seen in all of the promoters used in this study was not anticipated. While variations in promoter activity can be caused by a number of influencing factors, the variation in promoter activity observed here would imply a major contributing factor common to all plant expression cassettes tested. All promoter constructs generated in this study were based on the binary vector pCAMBIA2300. This vector contains the plant selection gene (NPTII) under the transcriptional control of the duplicated CaMV 35S promoter. This CaMV 35S promoter contains two enhancer domains that confer strong, constitutive expression of the selection gene and is located immediately upstream of the promoter-GUS fusion. During the course of this project, Yoo et al. (2005) reported that transgene expression is significantly affected when the expression cassette is located on the same T-DNA as the 35S enhancer. It was concluded, the trans-acting effects of the enhancer activate and control transgene expression causing irregular expression patterns. This phenomenon seems the most plausible reason for the constitutive expression profiles observed with the root- and seed-specific promoters assessed in this study. The expression from some promoters can be influenced by their cognate terminator sequences. Therefore, the Arabidopsis ARSK1, EIR1, ATS1 and ATS3 terminator sequences were isolated and incorporated into expression cassettes containing the GUS reporter gene under the control of their cognate promoters. Again, unrestricted GUS activity was displayed throughout transgenic plants transformed with these reporter gene fusions. As previously discussed constitutive GUS expression was most likely due to the trans-acting effect of the upstream CaMV 35S promoter in the selection cassette located on the same T-DNA. The results obtained in this study make it impossible to assess the influence matching terminators with their cognate promoters have on transgene expression profiles. The obvious future direction of research continuing from this study would be to transform pBIN-based promoter-GUS fusions (ie. constructs containing no CaMV 35S promoter driving the plant selection gene) into Arabidopsis in order to determine the true tissue specificity of these promoters and evaluate the effects of their cognate 3’ terminator sequences. Further, promoter truncations based around the cis-elements identified here may assist in determining whether these motifs are in fact involved in the overall activity of the promoter.

How to tell the difference between production and consumption : a case study in Doctor Who fandom

A television series is tagged with the label "cult" by the media, advertisers, and network executives when it is considered edgy or offbeat, when it appeals to nostalgia, or when it is considered emblematic of a particular subculture. By these criteria, almost any series could be described as cult. Yet certain programs exert an uncanny power over their fans, encouraging them to immerse themselves within a fictional world.In Cult Television leading scholars examine such shows as The X-Files; The Avengers; Doctor Who, Babylon Five; Star Trek; Xena, Warrior Princess; and Buffy the Vampire Slayer to determine the defining characteristics of cult television and map the contours of this phenomenon within the larger scope of popular culture.Contributors: Karen Backstein; David A. Black, Seton Hall U; Mary Hammond, Open U; Nathan Hunt, U of Nottingham; Mark Jancovich; Petra Kuppers, Bryant College; Philippe Le Guern, U of Angers, France; Alan McKee; Toby Miller, New York U; Jeffrey Sconce, Northwestern U; Eva ViethSara Gwenllian-Jones is a lecturer in television and digital media at Cardiff University and co-editor of Intensities: The Journal of Cult Media.Roberta E. Pearson is a reader in media and cultural studies at Cardiff University. She is the author of the forthcoming book Small Screen, Big Universe: Star Trek and Television.

Solar powered UAV for fire prevention and planning

This project aims to develop a methodology for designing and conducting a systems engineering analysis to build and fly continuously, day and night, propelled uniquely by solar energy for one week with a 0.25Kg payload consuming 0.5 watt without fuel or pollution. An airplane able to fly autonomously for many days could find many applications. Including coastal or border surveillance, atmospherical and weather research and prediction, environmental, forestry, agricultural, and oceanic monitoring, imaging for the media and real-estate industries, etc. Additional advantages of solar airplanes are their low cost and the simplicity with which they can be launched. For example, in the case of potential forest fire risks during a warm and dry period, swarms of solar airplanes, easily launched with the hand, could efficiently monitor a large surface, reporting rapidly any fire starts. This would allow a fast intervention and thus reduce the cost of such disaster, in terms of human and material losses. At higher dimension, solar HALE platforms are expected to play a major role as communication relays and could replace advantageously satellites in a near future.

Entering the era of post production the role visual effects will play in documentary

Creating an acceptance of Visual Effects (VFX) as an effective non-fiction communication tool has the potential to significantly boost return on investment for filmmakers producing documentary. Obtaining this acceptance does not necessarily mean rethinking the way documentary is defined, however, the need to address negative perceptions presently dominant within the production industry does exist; specifically, the misguided judgement that use of sequences which include visual effects discredits a filmmaker's attempt to represent reality. After completing a documentary utilising a traditional model of production for methodology, the question of how to increase this film's marketability is then examined by testing the specific assertion that Visual Effects is capable of increasing the level of appeal inherent within the documentary genre. Whilst this area of research is speculative, qualifying Visual Effects as an acceptable communication tool in non-fiction narratives will allow the documentary sector to benefit from increased production capabilities.

Planning and evaluation

Introduction: The core business of public health is to protect and promote health in the population. Public health planning is the means to maximise these aspirations. Health professionals develop plans to address contemporary health priorities as the evidence about changing patterns of mortality and morbidity is presented. Officials are also alert to international trends in patterns of disease that have the potential to affect the health of Australians. Integrated planning and preparation is currently underway involving all emergency health services, hospitals and population health units to ensure Australia's quick and efficient response to any major infectious disease outbreak, such as avian influenza (bird flu). Public health planning for the preparations for the Sydney Olympics and Paralympic Games in 2000 took almost three years. ‘Its major components included increased surveillance of communicable disease; presentations to sentinel emergency departments; medical encounters at Olympic venues; cruise ship surveillance; environmental and food safety inspections; bioterrorism surveillance and global epidemic intelligence’ (Jorm et al 2003, 102). In other words, the public health plan was developed to ensure food safety, hospital capacity, safe crowd control, protection against infectious diseases, and an integrated emergency and disaster plan. We have national and state plans for vaccinating children against infectious diseases in childhood; plans to promote dental health for children in schools; and screening programs for cervical, breast and prostate cancer. An effective public health response to a change in the distribution of morbidity and mortality requires planning. All levels of government plan for the public’s health. Local governments (councils) ensure healthy local environments to protect the public’s health. They plan parks for recreation, construct traffic-calming devices near schools to prevent childhood accidents, build shade structures and walking paths, and even embed drafts/chess squares in tables for people to sit and play. Environmental Health officers ensure food safety in restaurants and measure water quality. These public health measures attempt to promote the quality of life of residents. Australian and state governments produce plans that protect and promote health through various policy and program initiatives and innovations. To be effective, program plans need to be evaluated. However, building an integrated evaluation plan into a program plan is often forgotten, as planning and evaluation are seen as two distinct entities. Consequently, it is virtually impossible to measure, with any confidence, the extent to which a program has achieved its goals and objectives. This chapter introduces you to the concepts of public health program planning and evaluation. Case studies and reflection questions are presented to illustrate key points. As various authors use different terminology to describe the same concepts/actions of planning and evaluation, the glossary at the back of this book will help you to clarify the terms used in this chapter.

Emergency department nurses' understanding and experiences of implementing discharge planning

Aim: This paper is a report of a study conducted to describe emergency department nurses' understanding and experiences of implementing discharge planning. ---------- Background: Discharge planning in the emergency department is an important issue because of increased healthcare costs and greater emphasis on continuity of care. When executed as a collaborative process involving a multi-disciplinary team with the patient and family, discharge planning provides continuity of care for patients, less demand on hospitals, improvement in community services and in the services of other healthcare organizations. ---------- Method: The qualitative approach of phenomenography was used in this study. Thirty-two emergency department nurses were recruited between July and September 2005. Semi-structured interviews were conducted. ---------- Findings: From interviewees' descriptions of implementing discharge planning, six categories were established: implementing discharge planning as 'getting rid of my patients', completing routines, being involved in patient education, professionally accountable practice, autonomous practice and demonstrating professional emergency department nursing care. The referential meaning of implementing discharge planning 'in the outcome space' was the professional commitment to emergency department provision of effective discharge services. ---------- Conclusion: The results of this research contribute to knowledge of emergency department nurses' experience in the implementation of the discharge planning process. Key requirements for the provision of manageable discharge services both in Taiwan and worldwide highlighted by this study include adequate workloads, sufficient time, clear policies and standards of discharge planning and enhancement of professional commitment.

Discharge planning in the emergency department : a comprehensive approach

Discharge planning has become increasingly important, with current trends toward shorter hospital stays, increased health care costs, and more community-based health services. Effective discharge planning ensures the safety and ongoing care for patients,1 and it also benefits health care providers and organizations. It results in shorter hospital stays, fewer readmissions, higher access rates to post-hospitalization services, greater patient satisfaction with the discharge, and improved quality of life and continuity of care.[2] and [3] All acute care patients and their caregivers require some degree of preparation for discharge home—education about their health status, risks, and treatment; help setting health goals and maintaining a good level of self-care; information about community resources; and follow-up appointments and referrals to appropriate community health providers. Inadequate preparation exposes the patient to unnecessary risks of recurrence or complications of the acute complaint, neglect of nonacute comorbidities, mismanagement and side effects of medication, disruption of family and social life, emotional distress, and financial loss.[2], [3] and [4] The result may be re-presentation to the emergency department. It is noteworthy that up to 18% of ED presentations are revisits within 72 hours of the original visit5; many of these are considered preventable.6 It is a primary responsibility of nurses to ensure that patients return to the community adequately prepared and with appropriate support in place. Up to 65% of ED patients are discharged home from the emergency department,7 and the characteristics of the emergency department and its patient population make the provision of a high standard of discharge planning uniquely difficult. In addition, discharge planning is neglected in contemporary emergency nursing—there are no monographs devoted to the subject, and there is little published research. In this article 3 issues are explored: the importance of emergency nurses’ participation in the discharge-planning process, impediments to their participation; and strategies to improve discharge planning in the emergency department.

Alternative film production

Typically a film producer expects the director and actors to 'do their job' within a scheduled timeframe. Rather than expecting the creative principals to just deliver, a production model can be tailored to help this creative team produce successful outcomes. This research paper contrasts alternative production models with a traditional (or standard) production and presents possibilities for producers to emphasise the collaborative potential for their production.

Beyond the fashionable : strategic planning for critical information literacy education

The challenge for all educators is to fuse the learning of information literacy to an academic education in such a way that the outcome is systematic and sustainable learning for students. This challenge can be answered through long-term commitment to information literacy education bound to organisation-wide, renewable strategic planning and driven through systemic reform. This chapter seeks to explore the two sides of reforming information literacy education in an academic environment. Specifically, it will examine how one Australian university has undertaken the implementation of a rigorous strategic, systemic approach to information literacy learning and teaching.

Towards compatible and integrated airport and regional land use planning in Australia

Over the past decade privatised capital city airports in Australia have planned developed a range of non aviation commercial and retail land uses on airport land. Many surrounding municipalities consider this development in conflict with existing regional land use planning. Conversely airport operators are alarmed at continued urban consolidation and encroachment of incompatible regional development. Land use planning within and surrounding Australian capital city airports does not support compatible and integrated land use. It is currently a fragmented process due to: 1) current legislative and policy frameworks; 2) competing stakeholder priorities and interests; and 3) inadequate coordination and disjointed decision-making. This paper will examine privatised Australian airport development and consider three case studies to detail the context of airport and regional land use planning. A series of stakeholder workshops have served to inform the procedural dynamics and relationships between airport and regional decision-making. This exploratory research will assist in informing the knowledge gaps between aviation, airport development and broader urban land use policy. This paper will provide recommendations to enhance approaches to land use planning for airports and adjacent metropolitan regions in Australia and overseas.

Planning for survival

When the global financial crisis has its own acronym (the “GFC”), you know its going to be around for a while. This article looks at some of the risk/opportunity assessments you should take into consideration if you are planning for your property development business to still be around when the GFC is a thing of the past.

Guided chaos : path planning and control for a UAV-forced landing

In recent years, unmanned aerial vehicles (UAVs) have been widely used in combat, and their potential applications in civil and commercial roles are also receiving considerable attention by industry and the research community. There are numerous published reports of UAVs used in Earth science missions [1], fire-fighting [2], and border security [3] trials, with other speculative deployments, including applications in agriculture, communications, and traffic monitoring. However, none of these UAVs can demonstrate an equivalent level of safety to manned aircraft, particularly in the case of an engine failure, which would require an emergency or forced landing. This may be arguably the main factor that has prevented these UAV trials from becoming full-scale commercial operations, as well as restricted operations of civilian UAVs to only within segregated airspace.

Probabilistic world modeling for distributed team planning

This paper describes an application of decoupled probabilistic world modeling to achieve team planning. The research is based on the principle that the action selection mechanism of a member in a robot team can select an effective action if a global world model is available to all team members. In the real world, the sensors are imprecise, and are individual to each robot, hence providing each robot a partial and unique view about the environment. We address this problem by creating a probabilistic global view on each agent by combining the perceptual information from each robot. This probabilistic view forms the basis for selecting actions to achieve the team goal in a dynamic environment. Experiments have been carried out to investigate the effectiveness of this principle using custom-built robots for real world performance, in addition, to extensive simulation results. The results show an improvement in team effectiveness when using probabilistic world modeling based on perception sharing for team planning.

Health protection

Our understanding of how the environment can impact human health has evolved and expanded over the centuries, with concern and interest dating back to ancient times. For example, over 4000 years ago, a civilisation in northern India tried to protect the health of its citizens by constructing and positioning buildings according to strict building laws, by having bathrooms and drains, and by having paved streets with a sewerage system (Rosen 1993). In more recent times, the ‘industrial revolution’ played a dominant role in shaping the modern world, and with it the modern public health system. This era was signified by rapid progress in technology, the growth of transportation and the expansion of the market economy, which lead to the organisation of industry into a factory system. This meant that labour had to be brought to the factories and by the 1820s, poverty and social distress (including overcrowding and infrequent sewage and garbage disposal) was more widespread than ever. These circumstances, therefore, lead to the rise of the ‘sanitary revolution’ and the birth of modern public health (Rosen 1993). The sanitary revolution has also been described as constituting the beginning of the first wave of environmental concern, which continued until after World War 2 when major advances in engineering and chemistry substantially changed the face of industry, particularly the chemical sector. The second wave of environmental concern came in the mid to late 20th century and was dominated by the environmental or ecology movement. A landmark in this era was the 1962 publication of the book Silent Spring by Rachel Carson. This identified for the first time the dramatic effects on the ecosystem of the widespread use of the organochlorine pesticide, DDT. The third wave of environmental concern commenced in the 1980s and continues today. The accelerated rate of economic development, the substantial increase in the world population and the globalisation of trade have dramatically changed the production methods and demand for goods in both developed and developing countries. This has lead to the rise of ‘sustainable development’ as a key driver in environmental planning and economic development (Yassi et al 2001). The protection of health has, therefore, been a hallmark of human history and is the cornerstone of public health practice. This chapter introduces environmental health and how it is managed in Australia, including a discussion of the key generic management tools. A number of significant environmental health issues and how they are specifically managed are then discussed, and the chapter concludes by discussing sustainable development and its links with environmental health.

Contesting ‘sustainability’ in infrastructure planning : The debate of the public interest in Brisbane’s Boggo Road Busway development

Sustainable infrastructure demands that declared principles of sustainability are enacted in the processes of its implementation. However, a problem arises if the concept of sustainability is not thoroughly scrutinised in the planning process. The public interest could be undermined when the rhetoric of sustainability is used to substantiate a proposed plan. This chapter analyses the manifestation of sustainable development in the Boggo Road Busway Plan in Brisbane, Australia against the sustainability agenda set in the South East Queensland Regional and Transport Plans. Although the construction of the Busway was intended to improve public transport access in the region, its implementation drew significant environmental concerns. Local community groups contested the ‘sustainability’ concept deployed in Queensland’s infrastructure planning. Their challenges resulted in important concessions in the delivery of the Busway plan. This case demonstrates that principles of sustainable infrastructure should be measurable and that local communities be better informed in order to fulfil the public interest in regional planning.