Design and construction of an in-plant activation cassette for transgene expression and recombinant protein production in plants

Virus-based transgene expression systems have become particularly valuable for recombinant protein production in plants. The dual-module in-plant activation (INPACT) expression platform consists of a uniquely designed split-gene cassette incorporating the cis replication elements of Tobacco yellow dwarf geminivirus (TYDV) and an ethanol-inducible activation cassette encoding the TYDV Rep and RepA replication-associated proteins. The INPACT system is essentially tailored for recombinant protein production in stably transformed plants and provides both inducible and high-level transient transgene expression with the potential to be adapted to diverse crop species. The construction of a novel split-gene cassette, the inducible nature of the system and the ability to amplify transgene expression via rolling-circle replication differentiates this system from other DNA- and RNA-based virus vector systems used for stable or transient recombinant protein production in plants. Here we provide a detailed protocol describing the design and construction of a split-gene INPACT cassette, and we highlight factors that may influence optimal activation and amplification of gene expression in transgenic plants. By using Nicotiana tabacum, the protocol takes 6-9 months to complete, and recombinant proteins expressed using INPACT can accumulate to up to 10% of the leaf total soluble protein.

An instrument to assess the obesogenic environment of child care centers

OBJECTIVES To describe protocol and interobserver agreements of an instrument to evaluate nutrition and physical activity environments at child care. METHODS Interobserver data were collected from 9 child care centers, through direct observation and document review (17 observer pairs). RESULTS Mean agreement between observer pairs was 87.26% and 79.29% for the observation and document review, respectively. Items with lower agreement were primarily staff behavior, counting across the day/week, and policy classifications. CONCLUSIONS Although some revisions are required, the interobserver agreement for the environment and policy assessment and observation (EPAO instrument) appears to be quite good for assessing the nutrition and physical activity environment of child care centers.

Mortality following hip arthroplasty—inappropriate use of National Joint Registry (NJR) data

Mortality following hip arthroplasty is affected by a large number of confounding variables each of which must be considered to enable valid interpretation. Relevant variables available from the 2011 NJR data set were included in the Cox model. Mortality rates in hip arthroplasty patients were lower than in the age-matched population across all hip types. Age at surgery, ASA grade, diagnosis, gender, provider type, hip type and lead surgeon grade all had a significant effect on mortality. Schemper's statistic showed that only 18.98% of the variation in mortality was explained by the variables available in the NJR data set. It is inappropriate to use NJR data to study an outcome affected by a multitude of confounding variables when these cannot be adequately accounted for in the available data set.

“Best interests” and withholding and withdrawing life-sustaining treatment from an adult who lacks capacity in the parens patriae jurisdiction

Disputes about withholding and withdrawing life-sustaining treatment are increasingly coming before Australian Supreme Courts. Such cases are generally heard in the parens patriae jurisdiction where the test applied is what is in the patient’s “best interests”. However, the application of the “best interests” test, and its meaning, remains unclear in this context. To shed light on this emerging body of jurisprudence, this article analyses the Australian superior court decisions that consider an adult’s best interests in the context of decisions about life-sustaining treatment. We identify a number of themes from the current body of cases and consider how these themes may guide future decision-making. After then considering the law in the United Kingdom, we suggest an approach for assessing best interests that could be adopted by Australian Supreme Courts. We argue that the suggested approach will lead to a more structured and systematic decision-making process that better promotes the best interests of the patient.

Assessing the role of spatial correlations during collective cell spreading

Spreading cell fronts are essential features of development, repair and disease processes. Many mathematical models used to describe the motion of cell fronts, such as Fisher’s equation, invoke a mean–field assumption which implies that there is no spatial structure, such as cell clustering, present. Here, we examine the presence of spatial structure using a combination of in vitro circular barrier assays, discrete random walk simulations and pair correlation functions. In particular, we analyse discrete simulation data using pair correlation functions to show that spatial structure can form in a spreading population of cells either through sufficiently strong cell–to–cell adhesion or sufficiently rapid cell proliferation. We analyse images from a circular barrier assay describing the spreading of a population of MM127 melanoma cells using the same pair correlation functions. Our results indicate that the spreading melanoma cell populations remain very close to spatially uniform, suggesting that the strength of cell–to–cell adhesion and the rate of cell proliferation are both sufficiently small so as not to induce any spatial patterning in the spreading populations.

End of Life Decision Making and Palliative Care

1. An emergency department attendance represents an opportunity to set goals for care during the attendance and beyond. 2. End of life discussions and advance care planning assist early decision-making about treatment goals and end of life care. 3. Knowledge of the law assists decision-making at the end of life. 4. Not all dying patients require the skill set of a palliative care specialist but every dying patient will benefit from a palliative approach. 5. Palliative care does not preclude active treatment where the intent is understood by patient and family. 6. Failure to diagnose dying can compromise patient care. 7. The emergency department should foster close relationships with local specialist palliative care providers to improve and ensure timely access for patients and families and so that emergency staff have access to the knowledge and skills provided.

Asian international student attrition among Australian universities : are wellness programs the answer?

Education is one of Australia’s largest service-based exports. International students comprise approximately 24% of enrolments at Australian universities (Sawir, Marginson, Deumert, Nyland, & Ramia, 2008); and approximately 80% of these students are from the Asian region (Australian Federal Government, 2012). The financial cost of international student attrition for universities is significant. The Australian Federal Government Department of Education, Science and Training reports the attrition rates for first-year international undergraduate students ranged between 4% and 22.5% across all Australian Universities (2013). Academic, psychological, and sociocultural adjustments to a new environment can be challenging for international students. This process manifests from various stressors such as communication difficulties, adjustment to a new teaching style, new cultural norms and pressure on academic performance. These stressors result in an often overwhelming attempt to integrate and function effectively, and can consequently affect a student’s ability to meet academic requirements. The relationship between a student’s ability to successfully complete a higher education program is consistently related to a range of academic and non-academic factors. The role of specific Australian higher education institutions is vital in facilitating the continued education of Asian International students. Initiatives targeting an enhancement of modifiable lifestyle factors may have the potential to enhance a student’s ability to effectively and successfully transition into a lifestyle that facilitates their ability to adjust to the requirements of Australian universities. One possibility is the prospect of providing wellness programming, coaching and education targeting lifestyle behaviours for acculturation.

Cutting the Gordian knot of futility : a case for law reform on unilateral withholding and withdrawal of potentially life-sustaining treatment

In this paper, we propose law reform with respect to the unilateral withholding or withdrawal of potentially life-sustaining treatment in Australia and New Zealand. That is, where a doctor withholds or withdraws potentially life-sustaining treatment without consent from a patient or a patient’s substitute decision-maker (where the patient lacks capacity), or authorisation from a court or tribunal, or by operation of a statute or justifiable government or institutional policy. Our proposal is grounded in the core values that do (or should) underpin a regulatory framework on an issue such as this; these values are drawn from existing commitments made by Australia and New Zealand through legislation, the common law, and conventions and treaties. It is also grounded in a critical review of the law on unilateral withholding and withdrawal as well as the legal context within which this issue sits in Australasia. We argue that the current law is inconsistent with the core values and develop a proposal for a legal response to this issue that more closely aligns with the core values it is supposed to serve.

Health Law in Australia [Second Edition]

Health Law in Australia is the country’s leading text in this area and was the first book to deal with health law on a comprehensive national basis. In this important field that continues to give rise to challenges for society Health Law in Australia takes a logical, structured approach to explain the breadth of this area of law across all Australian jurisdictions. By covering all the major areas in this diverse field, Health Law in Australia enhances the understanding of the discipline as a whole. Beginning with an exploration of the general principles of health law, including chapters on “Negligence”, “Children and Consent to Medical Treatment”, and “Medical Confidentiality and Patient Privacy”, the book goes on to consider beginning-of-life and end-of-life issues before concluding with chapters on emerging areas in health law, such as biotechnology, genetic technologies and medical research. The contributing authors are national leaders who are specialists in these areas of health law and who can share with readers the results of their research. Health Law in Australia has been written for both legal and health audiences and is essential reading for undergraduate and postgraduate students, researchers and scholars in the disciplines of law, health and medicine, as well as health and legal practitioners, government departments and bodies in the health area, and private health providers.

Health law : scope, sources and forces

• For the purposes of this chapter, “health law” encapsulates regulation of the medical and health professions, the administration of health services and the maintenance of public health to the extent that it is connected to the provision of health services. • There are diverging views as to whether health law can be regarded as a discrete “area of law”. • Health law draws on other areas of law such as tort law, criminal law and family law. It also draws upon other disciplines, most notably medical and health ethics. • Social and economic forces have influenced the development and direction of health law, and these forces may become even more influential in the future. • The increasingly globalised world has implications for Australia's health systems and raises questions and creates commitments in respect of the international community. • Technological developments, including in respect of treatment, diagnosis and information management, create ongoing challenges for health law. • Patient rights, human rights and consumerism are increasingly key drivers in the development of health law. • Health law is significant to contemporary Australian society because of the gravity of the topics that fall within its ambit, its social relevance to so many aspects of human existence and endeavour, the important role it plays in protecting the vulnerable, and the extent to which it engages with fundamental principles of justice.

Adults who lack capacity : substitute decision-making

• Mechanisms to facilitate consent to healthcare for adults who lack capacity are necessary to ensure that these adults can lawfully receive appropriate medical treatment when needed. • In Australia, the common law plays only a limited role in this context, through its recognition of advance directives and through the parens patriae jurisdiction of superior courts. • Substitute decision-making for adults who lack capacity is facilitated primarily by guardianship and other related legislation. This legislation, which has been enacted in all Australian States and Territories, permits a range of decision-makers to make different types of healthcare decisions. • Substitute decision-makers can be appointed by the adult or by a guardianship or other tribunal. Where there is no appointed decision-maker, legislation generally empowers those close to the adult to make the relevant decision. Most Australian jurisdictions have also provided for statutory advance directives. • For the most serious of decisions, such as non-therapeutic sterilisations, consent can only be provided by a tribunal. Other decisions can generally be made by a range of substitute decision-makers. Some treatment, such as very minor treatment or that which is needed in an emergency, can be provided without consent. • Guardianship legislation generally establishes a set of principles and/or other criteria to guide healthcare decisions. Mechanisms have also been established to resolve disputes as to who is the appropriate decision-maker and how a decision should be made.

Withholding and withdrawing life-sustaining medical treatment

• At common law, a competent adult can refuse life-sustaining medical treatment, either contemporaneously or through an advance directive which will operate at a later time when the adult’s capacity is lost. • Legislation in most Australian jurisdictions also provides for a competent adult to complete an advance directive that refuses life-sustaining medical treatment. • At common law, a court exercising its parens patriae jurisdiction can consent to, or authorise, the withdrawal or withholding of life-sustaining medical treatment from an adult or child who lacks capacity if that is in the best interests of the person. A court may also declare that the withholding or withdrawal of treatment is lawful. • Guardianship legislation in all jurisdictions allows a substitute decision-maker, in an appropriate case, to refuse life-sustaining medical treatment for an adult who lacks capacity. • In terms of children, a parent may refuse life-sustaining medical treatment for his or her child if it is in the child’s best interests. • While a refusal of life-sustaining medical treatment by a competent child may be valid, this decision can be overturned by a court. • At common law and generally under guardianship statutes, demand for futile treatment need not be complied with by doctors.

Double effect and palliative care excuses

• The doctrine of double effect is an exception to the general rule that taking active steps that end life is unlawful. • The essence of the doctrine at common law is intention. • Hastening a patient’s death through palliative care will be lawful provided the primary intention is to relieve pain, and not cause death, even if that death is foreseen. • Some States have enacted legislative excuses that deal with the provision of palliative care. • These statutory excuses tend to be stricter than the common law as they impose other requirements in addition to having an appropriate intent, such as adherence to some level of recognised medical practice.

The French polisher and the unsentimental interior

In January 2011 a swollen Brisbane River broke its banks flooding riverside houses and buildings. The river’s water spread and rose up through storm water drains inundating some 20 000 houses in low-lying land. As the water receded those residents affected by the floods returned to their homes to assess the damage. While some people breathed a sigh of relief others were devastated by the overwhelming damage to their homes and personal belongings. Over the next few weeks the landscape of Brisbane was altered not merely by the mud and debris left by the torrent of water, but by the piles of domestic contents occupying Brisbane streets. Beds, toys, cabinets, plasterboard, tiles and household furniture lined curbsides waiting for collection. Later they would accumulate in public parks and sports centres to await disposal, momentarily creating an unsettling landscape of discarded domestic interiors. While most houses remained standing the heart breaking repercussions were evident in their interiority. Thousands of volunteers flocked to help those affected by the floods to purge the damage left by the water – removing wall and floor linings, discarding furniture and spoilt belongings. In her paper on Hurricane Katrina, Julieanna Preston wrote, ‘What anthropological evidence would we find as we followed their migration – heaps left by the side of the road, the physical weight overcoming the personal value…’ For many of the post flood restored homes and buildings entire interiors have been replaced, eradicating any trace of the significant event that disturbed them only months earlier. There were artifacts that would have survived the floods - furniture of solid timber – these were discarded and with them the patina that marked an important event in history. The patina is beyond technological reproducibility, and as Walter Benjamin writes, this being the whole premise of genuineness. It is the role of the French Polisher to maintain the true wear of the artefact for it is the patina that is most valuable in its ability to narrate the history of a piece. In 2012 two separate exhibitions in Brisbane will take place to display a selected collection of flood-damaged artefacts. This orchestrated way to commemorate the damage left by floods may be a method to compensate for the haste in which the damage was purged from the city. This need for exhibiting damaged artifacts illustrates Andreas Huyssen’s point that "…today memory is understood as a mode of re-presentation and as belonging to the present." This research looks at the dying trade of the French Polisher through conversations and a visual study of flood damaged furniture. The research also investigates the personal loss of artifacts through intimate stories shared by flood victims. This paper seeks to understand why so much was discarded and celebrate what remains.

Photophysical characterization of light-emitting poly(indenofluorene)s

Time-resolved photoluminescence spectroscopy experiments of three poly(2,8-indenofluorene) derivatives bearing different pendant groups are presented. A comparison of the photophysical properties of dilute solutions and thin films provides information on the chemical purity of the materials. The photophysical properties of poly(2,8-indenofluorene)s are correlated with the morphological characteristics of their corresponding films. Wide-angle X-ray scattering experiments reveal the order in these materials at the molecular level. The spectroscopic results confirm the positive impact of a new synthetic approach on the spectral purity of the poly(indenofluorene)s. It is concluded that complete side-chain substitution of the bridgehead carbon atoms C-6 and C-12 in the indenofluorene unit, prior to indenofluorene ring formation, reduces the probability of keto formation. Due to the intrinsic chemical purity of the arylated derivative, identification of a long-delayed spectral feature, other than the known keto band, is possible in the case of thin films. Controlled doping experiments on the arylated derivative with trace amounts of an indenofluorene-monoketone provide quantitative information on the rates of two major photophysical processes, namely, singlet photoluminescence emission and singlet photoluminescence quenching. These results allow the determination of the minimum keto concentration that can affect the intrinsic photophysical properties of this polymer. The data suggest that photoluminescence quenching operates in the doped films according to the Stern-Volmer formalism.