Refining animal models in fracture research: Seeking consensus in optimising both animal welfare and scientific validity for appropriate biomedical use

Background In an attempt to establish some consensus on the proper use and design of experimental animal models in musculoskeletal research, AOVET (the veterinary specialty group of the AO Foundation) in concert with the AO Research Institute (ARI), and the European Academy for the Study of Scientific and Technological Advance, convened a group of musculoskeletal researchers, veterinarians, legal experts, and ethicists to discuss, in a frank and open forum, the use of animals in musculoskeletal research. Methods The group narrowed the field to fracture research. The consensus opinion resulting from this workshop can be summarized as follows: Results & Conclusion Anaesthesia and pain management protocols for research animals should follow standard protocols applied in clinical work for the species involved. This will improve morbidity and mortality outcomes. A database should be established to facilitate selection of anaesthesia and pain management protocols for specific experimental surgical procedures and adopted as an International Standard (IS) according to animal species selected. A list of 10 golden rules and requirements for conduction of animal experiments in musculoskeletal research was drawn up comprising 1) Intelligent study designs to receive appropriate answers; 2) Minimal complication rates (5 to max. 10%); 3) Defined end-points for both welfare and scientific outputs analogous to quality assessment (QA) audit of protocols in GLP studies; 4) Sufficient details for materials and methods applied; 5) Potentially confounding variables (genetic background, seasonal, hormonal, size, histological, and biomechanical differences); 6) Post-operative management with emphasis on analgesia and follow-up examinations; 7) Study protocols to satisfy criteria established for a "justified animal study"; 8) Surgical expertise to conduct surgery on animals; 9) Pilot studies as a critical part of model validation and powering of the definitive study design; 10) Criteria for funding agencies to include requirements related to animal experiments as part of the overall scientific proposal review protocols. Such agencies are also encouraged to seriously consider and adopt the recommendations described here when awarding funds for specific projects. Specific new requirements and mandates related both to improving the welfare and scientific rigour of animal-based research models are urgently needed as part of international harmonization of standards.

Destination positioning opportunities using personal values : elicited through the Repertory Test with Laddering Analysis

While in many travel situations there is an almost limitless range of available destinations, travellers will usually only actively consider two to six in their decision set. One of the greatest challenges facing destination marketers is positioning their destination, against the myriad of competing places that offer similar features, into consumer decision sets. Since positioning requires a narrow focus, marketing communications must present a succinct and meaningful proposition, the selection of which is often problematic for destination marketing organisations (DMO), which deal with a diverse and often eclectic range of attributes in addition to self-interested and demanding stakeholders who have interests in different market segments. This paper reports the application of two qualitative techniques used to explore the range of cognitive attributes, consequences and personal values that represent potential positioning opportunities in the context of short break holidays. The Repertory Test is an effective technique for understanding the salient attributes used by a traveller to differentiate destinations, and Laddering Analysis enables the researcher to explore the smaller set of consequences and personal values guiding such decision making. A key finding of the research was that while individuals might vary in their repertoire of salient attributes, there was a commonality of shared consequences and values. This has important implications for DMOs, since a brand positioning theme that is based on a value will subsume multiple and diverse attributes. It is posited that such a theme will appeal to a broader range of travellers, as well as appease a greater number of destination stakeholders, than would an attribute based theme.

CDKN2A mutation in a non-FAMMM kindred with cancers at multiple sites results in a functionally abnormal protein

The CDKN2A gene encodes p16 (CDKN2A), a cell-cycle inhibitor protein which prevents inappropriate cell cycling and, hence, proliferation. Germ-line mutations in CDKN2A predispose to the familial atypical multiple-mole melanoma (FAMMM) syndrome but also have been seen in rare families in which only 1 or 2 individuals are affected by cutaneous malignant melanoma (CMM). We therefore sequenced exons 1alpha and 2 of CDKN2A using lymphocyte DNA isolated from index cases from 67 families with cancers at multiple sites, where the patterns of cancer did not resemble those attributable to known genes such as hMLH1, hMLH2, BRCA1, BRCA2, TP53 or other cancer susceptibility genes. We found one mutation, a mis-sense mutation resulting in a methionine to isoleucine change at codon 53 (M531) of exon 2. The individual tested had developed 2 CMMs but had no dysplastic nevi and lacked a family history of dysplastic nevi or CMM. Other family members had been diagnosed with oral cancer (2 persons), bladder cancer (1 person) and possibly gall-bladder cancer. While this mutation has been reported in Australian and North American melanoma kindreds, we did not observe it in 618 chromosomes from Scottish and Canadian controls. Functional studies revealed that the CDKN2A variant carrying the M531 change was unable to bind effectively to CDK4, showing that this mutation is of pathological significance. Our results have confirmed that CDKN2A mutations are not limited to FAMMM kindreds but also demonstrate that multi-site cancer families without melanoma are very unlikely to contain CDKN2A mutations.

p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

p53 is the central member of a critical tumor suppressor pathway in virtually all tumor types, where it is silenced mainly by missense mutations. In melanoma, p53 predominantly remains wild type, thus its role has been neglected. To study the effect of p53 on melanocyte function and melanomagenesis, we crossed the 'high-p53'Mdm4+/- mouse to the well-established TP-ras0/+ murine melanoma progression model. After treatment with the carcinogen dimethylbenzanthracene (DMBA), TP-ras0/+ mice on the Mdm4+/- background developed fewer tumors with a delay in the age of onset of melanomas compared to TP-ras0/+ mice. Furthermore, we observed a dramatic decrease in tumor growth, lack of metastasis with increased survival of TP-ras0/+: Mdm4+/- mice. Thus, p53 effectively prevented the conversion of small benign tumors to malignant and metastatic melanoma. p53 activation in cultured primary melanocyte and melanoma cell lines using Nutlin-3, a specific Mdm2 antagonist, supported these findings. Moreover, global gene expression and network analysis of Nutlin-3-treated primary human melanocytes indicated that cell cycle regulation through the p21WAF1/CIP1 signaling network may be the key anti-melanomagenic activity of p53.

Profluorescent nitroxides as sensitive probes of oxidative change and free radical reactions

This paper presents a review on the use of tethered nitroxide–fluorophore molecules as probes of oxidative change and free radical generation and reaction. The proximity of the nitroxide free radical to the fluorophore suppresses the normal fluorescence emission process. Nitroxide free radical scavenging, metabolism or redox chemistry return the system to its natural fluorescent state and so these tethered nitroxide–fluorophore molecules are described as being profluorescent. A survey of profluorescent nitroxides found in the literature is provided as well as background on the mechanism of action and applications of these compounds as fluorometric probes within the fields of biological, materials and environmental sciences.

Evolving noisy oscillatory dynamics in genetic regulatory networks

We introduce a genetic programming (GP) approach for evolving genetic networks that demonstrate desired dynamics when simulated as a discrete stochastic process. Our representation of genetic networks is based on a biochemical reaction model including key elements such as transcription, translation and post-translational modifications. The stochastic, reaction-based GP system is similar but not identical with algorithmic chemistries. We evolved genetic networks with noisy oscillatory dynamics. The results show the practicality of evolving particular dynamics in gene regulatory networks when modelled with intrinsic noise.

Juvenile justice and truancy legislation : The move towards parental responsibilisation in Queensland

Much current Queensland media rhetoric, government policy and legislation on truancy and youth justice appears to be based on ideas of responsibilisation – of sheeting responsibility for children’s behaviour back onto their parents. This article examines the evidence of parental responsibility provisions in juvenile justice and truancy legislation in Queensland and the drivers behind this approach. It considers recent legislative initiatives as part of an international trend toward making parents ‘responsible’ for the wrongs of their children. It identifies the parental responsibility rhetoric appearing in recent ministerial statements and associated media reports. It then asks the questions – are these legislative provisions being enforced? And if so, are they successful? Are they simply adding to the administrative burdens placed on teachers and schools, and the socioeconomic burdens placed on already disadvantaged parents? Parental responsibility provisions have been discussed at length in the context of juvenile offending and research suggests that punishing parents for the acts of their children does not decrease delinquency. The paper asks how, as a society, we intend to evaluate these punitive measures against parents?

‘Expected satiety’ changes hunger and fullness in the inter-meal interval

Previously, we have shown that foods differ markedly in the satiety that they are expected to confer (compared calorie-for-calorie). In the present study we tested the hypothesis that ‘expected satiety’ plays a causal role in the satiety that is experienced after a food has been consumed. Before lunch, participants (N = 32) were shown the ingredients of a fruit smoothie. Half were shown a small portion of fruit and half were shown a large portion. Participants then assessed the expected satiety of the smoothie and provided appetite ratings, before, and for three hours after its consumption. As anticipated, expected satiety was significantly higher in the ‘large portion’ condition. Moreover, and consistent with our hypothesis, participants reported significantly less hunger and significantly greater fullness in the large portion condition. Importantly, this effect endured throughout the test period (for three hours). Together, these findings confirm previous reports indicating that beliefs and expectations can have marked effects on satiety and they show that this effect can persist well into the inter-meal interval. Potential explanations are discussed, including the prospect that satiety is moderated by memories of expected satiety that are encoded around the time that a meal is consumed.

Parental responsibility and Australian legislative attempts to curb truancy

Statistical and anecdotal evidence suggests that truancy is a significant problem for Australian schools. This paper considers the efficacy of legislative attempts to curb truancy, focussing in particular on the Queensland experience. Both Queensland legislation and the Commonwealth Improving School Enrolment and Attendance Through Welfare reform Measure (SEAM) pilot program are explained and evaluated. The paper considers in particular the utility of parental responsibility strategies as a response to truancy - under the Education (General Provisions) Act 12006 (Queensland) parents of persistent truants may be prosecuted and fined; under the SEAM initiative parents may have their social security payments suspended. Despite the availability of these seemingly draconian penalties, there is a reluctance, in practice, to hold parents accountable. The paper attempts to explain this reluctance and asks whether parental responsibility legislation can deliver a solution to truancy.

Vision based control for fixed wing UAVs inspecting locally linear infrastructure using skid-to-turn maneuvers

The following paper proposes a novel application of Skid-to-Turn maneuvers for fixed wing Unmanned Aerial Vehicles (UAVs) inspecting locally linear infrastructure. Fixed wing UAVs, following the design of manned aircraft, traditionally employ Bank-to-Turn maneuvers to change heading and thus direction of travel. Commonly overlooked is the effect these maneuvers have on downward facing body fixed sensors, which as a result of bank, point away from the feature during turns. By adopting Skid-to-Turn maneuvers, the aircraft is able change heading whilst maintaining wings level flight, thus allowing body fixed sensors to maintain a downward facing orientation. Eliminating roll also helps to improve data quality, as sensors are no longer subjected to the swinging motion induced as they pivot about an axis perpendicular to their line of sight. Traditional tracking controllers that apply an indirect approach of capturing ground based data by flying directly overhead can also see the feature off center due to steady state pitch and roll required to stay on course. An Image Based Visual Servo controller is developed to address this issue, allowing features to be directly tracked within the image plane. Performance of the proposed controller is tested against that of a Bank-to-Turn tracking controller driven by GPS derived cross track error in a simulation environment developed to simulate the field of view of a body fixed camera.

Path planning using surface shape and ground properties

This paper presents a path planning technique for ground vehicles that accounts for the dynamics of the vehicle, the topography of the terrain and the wheel/ground interaction properties such as friction. The first two properties can be estimated using well known sensors and techniques, but the third is not often estimated even though it has a significant effect on the motion of a high-speed vehicle. We introduce a technique which allows the estimation of wheel slip from which frictional parameters can be inferred. We present simulation results which show the importance of modelling topography and ground properties and experimental results which show how ground properties can be estimated along a 350m outdoor traverse.