Development of zonal cartilage constructs : effects of chondrocyte subpopulation, compressive stimulation, and culture models

Articular cartilage is a highly resilient tissue located at the ends of long bones. It has a zonal structure, which has functional significance in load-bearing. Cartilage does not spontaneously heal itself when damaged, and untreated cartilage lesions or age-related wear often lead to osteoarthritis (OA). OA is a degenerative condition that is highly prevalent, age-associated, and significantly affects patient mobility and quality of life. There is no cure for OA, and patients usually resort to replacing the biological joint with an artificial prosthesis. An alternative approach is to dynamically regenerate damaged or diseased cartilage through cartilage tissue engineering, where cells, materials, and stimuli are combined to form new cartilage. However, despite extensive research, major limitations remain that have prevented the wide-spread application of tissue-engineered cartilage. Critically, there is a dearth of information on whether autologous chondrocytes obtained from OA patients can be used to successfully generate cartilage tissues with structural hierarchy typically found in normal articular cartilage. I aim to address these limitations in this thesis by showing that chondrocyte subpopulations isolated from macroscopically normal areas of the cartilage can be used to engineer stratified cartilage tissues and that compressive loading plays an important role in zone-dependent biosynthesis of these chondrocytes. I first demonstrate that chondrocyte subpopulations from the superficial (S) and middle/deep (MD) zones of OA cartilage are responsive to compressive stimulation in vitro, and that the effect of compression on construct quality is zone-dependent. I also show that compressive stimulation can influence pericelluar matrix production, matrix metalloproteinase secretion, and cytokine expression in zonal chondrocytes in an alginate hydrogel model. Subsequently, I focus on recreating the zonal structure by forming layered constructs using the alginate-released chondrocyte (ARC) method either with or without polymeric scaffolds. Resulting zonal ARC constructs had hyaline morphology, and expressed cartilage matrix molecules such as proteoglycans and collagen type II in both scaffold-free and scaffold-based approaches. Overall, my findings demonstrate that chondrocyte subpopulations obtained from OA joints respond sensitively to compressive stimulation, and are able to form cartilaginous constructs with stratified organization similar to native cartilage using the scaffold-free and scaffold-based ARC technique. The ultimate goal in tissue engineering is to help provide improved treatment options for patients suffering from debilitating conditions such as OA. Further investigations in developing functional cartilage replacement tissues using autologous chondrocytes will bring us a step closer to improving the quality of life for millions of OA patients worldwide.

Why do policy decision-makers opt for command and control environmental regulation? An economic analysis with special reference to Sri Lanka

This chapter examines why policy decision-makers opt for command and control environmental regulation despite the availability of a plethora of market-based instruments which are more efficient and cost-effective. Interestingly, Sri Lanka has adopted a wholly command and control system, during both the pre and post liberalisation economic policies. This chapter first examines the merits and demerits of command and control and market-based approaches and then looks at Sri Lanka’s extensive environmental regulatory framework. The chapter then examines the likely reasons as to why the country has gone down the path of inflexible regulatory measures and has become entrenched in them. The various hypotheses are discussed and empirical evidence is provided. The chapter also discusses the consequences of an environmentally slack economy and policy implications stemming from adopting a wholly regulatory approach. The chapter concludes with a discussion of the main results.

Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. We therefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor-3alpha mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration.

Circulating tumour cells, their role in metastasis and their clinical utility in lung cancer

Circulating tumour cells (CTCs) have attracted much recent interest in cancer research as a potential biomarker and as a means of studying the process of metastasis. It has long been understood that metastasis is a hallmark of malignancy, and conceptual theories on the basis of metastasis from the nineteenth century foretold the existence of a tumour "seed" which is capable of establishing discrete tumours in the "soil" of distant organs. This prescient "seed and soil" hypothesis accurately predicted the existence of CTCs; microscopic tumour fragments in the blood, at least some of which are capable of forming metastases. However, it is only in recent years that reliable, reproducible methods of CTC detection and analysis have been developed. To date, the majority of studies have employed the CellSearch™ system (Veridex LLC), which is an immunomagnetic purification method. Other promising techniques include microfluidic filters, isolation of tumour cells by size using microporous polycarbonate filters and flow cytometry-based approaches. While many challenges still exist, the detection of CTCs in blood is becoming increasingly feasible, giving rise to some tantalizing questions about the use of CTCs as a potential biomarker. CTC enumeration has been used to guide prognosis in patients with metastatic disease, and to act as a surrogate marker for disease response during therapy. Other possible uses for CTC detection include prognostication in early stage patients, identifying patients requiring adjuvant therapy, or in surveillance, for the detection of relapsing disease. Another exciting possible use for CTC detection assays is the molecular and genetic characterization of CTCs to act as a "liquid biopsy" representative of the primary tumour. Indeed it has already been demonstrated that it is possible to detect HER2, KRAS and EGFR mutation status in breast, colon and lung cancer CTCs respectively. In the course of this review, we shall discuss the biology of CTCs and their role in metastagenesis, the most commonly used techniques for their detection and the evidence to date of their clinical utility, with particular reference to lung cancer.

Finding additional semantic entity information for search engines

Entity-oriented search has become an essential component of modern search engines. It focuses on retrieving a list of entities or information about the specific entities instead of documents. In this paper, we study the problem of finding entity related information, referred to as attribute-value pairs, that play a significant role in searching target entities. We propose a novel decomposition framework combining reduced relations and the discriminative model, Conditional Random Field (CRF), for automatically finding entity-related attribute-value pairs from free text documents. This decomposition framework allows us to locate potential text fragments and identify the hidden semantics, in the form of attribute-value pairs for user queries. Empirical analysis shows that the decomposition framework outperforms pattern-based approaches due to its capability of effective integration of syntactic and semantic features.

Non-intrusive continuous user behaviour analysis using computerized systems

There are different ways to authenticate humans, which is an essential prerequisite for access control. The authentication process can be subdivided into three categories that rely on something someone i) knows (e.g. password), and/or ii) has (e.g. smart card), and/or iii) is (biometric features). Besides classical attacks on password solutions and the risk that identity-related objects can be stolen, traditional biometric solutions have their own disadvantages such as the requirement of expensive devices, risk of stolen bio-templates etc. Moreover, existing approaches provide the authentication process usually performed only once initially. Non-intrusive and continuous monitoring of user activities emerges as promising solution in hardening authentication process: iii-2) how so. behaves. In recent years various keystroke dynamic behavior-based approaches were published that are able to authenticate humans based on their typing behavior. The majority focuses on so-called static text approaches, where users are requested to type a previously defined text. Relatively few techniques are based on free text approaches that allow a transparent monitoring of user activities and provide continuous verification. Unfortunately only few solutions are deployable in application environments under realistic conditions. Unsolved problems are for instance scalability problems, high response times and error rates. The aim of this work is the development of behavioral-based verification solutions. Our main requirement is to deploy these solutions under realistic conditions within existing environments in order to enable a transparent and free text based continuous verification of active users with low error rates and response times.

Optimally increasing secure connectivity in multihop wireless ad hoc networks

We consider the problem of how to maximize secure connectivity of multi-hop wireless ad hoc networks after deployment. Two approaches, based on graph augmentation problems with nonlinear edge costs, are formulated. The first one is based on establishing a secret key using only the links that are already secured by secret keys. This problem is in NP-hard and does not accept polynomial time approximation scheme PTAS since minimum cutsets to be augmented do not admit constant costs. The second one is based of increasing the power level between a pair of nodes that has a secret key to enable them physically connect. This problem can be formulated as the optimal key establishment problem with interference constraints with bi-objectives: (i) maximizing the concurrent key establishment flow, (ii) minimizing the cost. We show that both problems are NP-hard and MAX-SNP (i.e., it is NP-hard to approximate them within a factor of 1 + e for e > 0 ) with a reduction to MAX3SAT problem. Thus, we design and implement a fully distributed algorithm for authenticated key establishment in wireless sensor networks where each sensor knows only its one- hop neighborhood. Our witness based approaches find witnesses in multi-hop neighborhood to authenticate the key establishment between two sensor nodes which do not share a key and which are not connected through a secure path.

Monitoring android for collaborative anomaly detection : a first architectural draft

Our daily lives become more and more dependent upon smartphones due to their increased capabilities. Smartphones are used in various ways, e.g. for payment systems or assisting the lives of elderly or disabled people. Security threats for these devices become more and more dangerous since there is still a lack of proper security tools for protection. Android emerges as an open smartphone platform which allows modification even on operating system level and where third-party developers first time have the opportunity to develop kernel-based low-level security tools. Android quickly gained its popularity among smartphone developers and even beyond since it bases on Java on top of "open" Linux in comparison to former proprietary platforms which have very restrictive SDKs and corresponding APIs. Symbian OS, holding the greatest market share among all smartphone OSs, was even closing critical APIs to common developers and introduced application certification. This was done since this OS was the main target for smartphone malwares in the past. In fact, more than 290 malwares designed for Symbian OS appeared from July 2004 to July 2008. Android, in turn, promises to be completely open source. Together with the Linux-based smartphone OS OpenMoko, open smartphone platforms may attract malware writers for creating malicious applications endangering the critical smartphone applications and owners privacy. Since signature-based approaches mainly detect known malwares, anomaly-based approaches can be a valuable addition to these systems. They base on mathematical algorithms processing data that describe the state of a certain device. For gaining this data, a monitoring client is needed that has to extract usable information (features) from the monitored system. Our approach follows a dual system for analyzing these features. On the one hand, functionality for on-device light-weight detection is provided. But since most algorithms are resource exhaustive, remote feature analysis is provided on the other hand. Having this dual system enables event-based detection that can react to the current detection need. In our ongoing research we aim to investigates the feasibility of light-weight on-device detection for certain occasions. On other occasions, whenever significant changes are detected on the device, the system can trigger remote detection with heavy-weight algorithms for better detection results. In the absence of the server respectively as a supplementary approach, we also consider a collaborative scenario. Here, mobile devices sharing a common objective are enabled by a collaboration module to share information, such as intrusion detection data and results. This is based on an ad-hoc network mode that can be provided by a WiFi or Bluetooth adapter nearly every smartphone possesses.

I know, but I don’t care : how awareness of Queensland’s drug driving testing methods impact upon perceptions of deterrence and offending behaviours

An increasing body of research is highlighting the involvement of illicit drugs in many road fatalities. Deterrence theory has been a core conceptual framework underpinning traffic enforcement as well as interventions designed to reduce road fatalities. Essentially the effectiveness of deterrence-based approaches is predicated on perceptions of certainty, severity, and swiftness of apprehension. However, much less is known about how the awareness of legal sanctions can impact upon the effectiveness of deterrence mechanisms and whether promoting such detection methods can increase the deterrent effect. Nevertheless, the implicit assumption is that individuals aware of the legal sanctions will be more deterred. This study seeks to explore how awareness of the testing method impacts upon the effectiveness of deterrence-based interventions and intentions to drug drive again in the future. In total, 161 participants who reported drug driving in the previous six months took part in the current study. The results show that awareness of testing had a small effect upon increasing perceptions of the certainty of apprehension and severity of punishment. However, awareness was not a significant predictor of intentions to drug drive again in the future. Importantly, higher levels of drug use were a significant predictor of intentions to drug drive in the future. Whilst awareness does have a small effect on deterrence variables, the influence of levels of drug use seems to reduce any deterrent effect.

Communicating personal amnesty : a dismodernist approach to arts intersections between disability, personal stories and well-being

This thesis investigates the role of narrative devices in the process of improving an individual’s psychological and physiological experience of health and well-being using two methods of inquiry: a theoretical research project and a comparative analysis of two case studies. Through these two approaches the research examines how the health status of people experiencing disability can be re-positioned and re-designed to develop creative, narrative-based approaches to strengthen communication between the mainstream community and those marginalised by pathological, social and biological illness-centric policy. The theoretical section of the thesis examines two different, but complementary bodies of research: health and well-being, and narrative reconstruction. By invoking Antonovksy’s (1985a) theory of salutogenesis and Davis’s (2002) theory of dismodernism, the study examines the role of language and narrative in the defining of health in social, pathological and ableist spheres. The research positions health and well-being as disparate from historical and contemporary readings of illness and disability and presents literature to support the potential to improve health well-being through a creative re-narration of the experience of disability. The research examines the theoretical concepts of resilience, autonomy and social inclusion through a detailed examination of narratology and the amnesty narrative. The study links these theoretical approaches to a practical Arts-Health intersection program developed for the research project called Communicating Personal Amnesty. Through a comparative analysis of a Pilot Study and Major Case study, the research presents findings derived from theory-building participatory action research showing the efficacy of the program. The research provides a detailed analysis of key narrative structures through a variety of experimental methodological approaches to encourage an important dialogue between the creative components of the thesis and the more traditional health-based academic critique. The research is an example of emergent translational health methodologies, in disability studies.

Evaluating Communication for Development : A Framework for Social Change

Evaluating Communication for Development presents a comprehensive framework for evaluating communication for development (C4D). This framework combines the latest thinking from a number of fields in new ways. It critiques dominant instrumental, accountability-based approaches to development and evaluation and offers an alternative holistic, participatory, mixed methods approach based on systems and complexity thinking and other key concepts. It maintains a focus on power, gender and other differences and social norms. The authors have designed the framework as a way to focus on achieving sustainable social change and to continually improve and develop C4D initiatives. The benefits and rigour of this approach are supported by examples and case studies from a number of action research and evaluation capacity development projects undertaken by the authors over the past fifteen years. Building on current arguments within the fields of C4D and development, the authors reinforce the case for effective communication being a central and vital component of participatory forms of development, something that needs to be appreciated by decision makers. They also consider ways of increasing the effectiveness of evaluation capacity development from grassroots to management level in the development context, an issue of growing importance to improving the quality, effectiveness and utilisation of monitoring and evaluation studies in this field. The book includes a critical review of the key approaches, methodologies and methods that are considered effective for planning evaluation, assessing the outcomes of C4D, and engaging in continuous learning. This rigorous book is of immense theoretical and practical value to students, scholars, and professionals researching or working in development, communication and media, applied anthropology, and evaluation and program planning.

Evaluation in health outcomes research : linking theories, methodologies and practice in health promotion.

Health outcomes research has developed as a means of evaluating the effectiveness of health care interventions and as an approach to informing resource allocation. The use of a health outcomes approach in health promotion has made increasing demands on evaluation methodologies to demonstrate program effectiveness. However, criticism of the contribution of health promotion to outcomes research has made several assumptions about the use of qualitative methodologies and the content of program objectives largely derived from a biomedical approach. In contrast to the measurement of biomedical interventions in clinical health care, health promotion practice involves social phenomena, wide-reaching cultural, psychological, political and ideological problems and issues. The integration of methodologies of health promotion evaluation will inform further conceptualisation of the health outcomes approach with the differentiation of three types of outcomes: health development outcomes; social health outcomes; and biomedical health outcomes. It is concluded that this differentiation moves away from dualist concepts that advocate the replacement of goals and targets with regional and locally based approaches. Rather, the future direction for health promotion evaluation needs to employ a framework that elaborates multiple methodologies and approaches necessary for establishing what relationships exist between morbidity, mortality, health advancement and equity.

Augmented Reality Process Modelling Tool

Video presented as part of BPM2011 demonstration(France). In this video we show a prototype BPMN process modelling tool which uses Augmented Reality techniques to increase the sense of immersion when editing a process model. The avatar represents a remotely logged in user, and facilitates greater insight into the editing actions of the collaborator than present 2D web-based approaches in collaborative process modelling. We modified the Second Life client to integrate the ARToolkit in order to support pattern-based AR.

Establishment and characterisation of an open mini-thoracotomy surgical approach to an ovine thoracic spine fusion model

Background A large animal model is required for assessment of minimally invasive, tissue engineering based approaches to thoracic spine fusion, with relevance to deformity correction surgery for human adolescent idiopathic scoliosis. Here we develop a novel open mini–thoracotomy approach in an ovine model of thoracic interbody fusion which allows assessment of various fusion constructs, with a focus on novel, tissue engineering based interventions. Methods The open mini-thoracotomy surgical approach was developed through a series of mock surgeries, and then applied in a live sheep study. Customized scaffolds were manufactured to conform with intervertebral disc space clearances required of the study. Twelve male Merino sheep aged 4 to 6 years and weighing 35 – 45 kg underwent the abovementioned procedure and were divided into two groups of six sheep at survival timelines of 6 and 12 months. Each sheep underwent a 3-level discectomy (T6/7, T8/9 and T10/11) with randomly allocated implantation of a different graft substitute at each of the three levels; (i) polycaprolactone (PCL) based scaffold plus 0.54μg rhBMP-2, (ii) PCL-based scaffold alone or (iii) autograft. The sheep were closely monitored post- operatively for signs of pain (i.e. gait abnormalities/ teeth gnawing/ social isolation). Fusion assessments were conducted post-sacrifice using Computed Tomography and hard-tissue histology. All scientific work was undertaken in accordance with the study protocol has been approved by the Institute's committee on animal research. Results. All twelve sheep were successfully operated on and reached the allotted survival timelines, thereby demonstrating the feasibility of the surgical procedure and post-operative care. There were no significant complications and during the post-operative period the animals did not exhibit marked signs of distress according to the described assessment criteria. Computed Tomographic scanning demonstrated higher fusion grades in the rhBMP-2 plus PCL-based scaffold group in comparison to either PCL-based scaffold alone or autograft. These results were supported by histological evaluation of the respective groups. Conclusion. This novel open mini-thoracotomy surgical approach to the ovine thoracic spine represents a safe surgical method which can reproducibly form the platform for research into various spine tissue engineered constructs (TEC) and their fusion promoting properties.

Program-level support for protecting an application from untrustworthy components

Many software applications extend their functionality by dynamically loading executable components into their allocated address space. Such components, exemplified by browser plugins and other software add-ons, not only enable reusability, but also promote programming simplicity, as they reside in the same address space as their host application, supporting easy sharing of complex data structures and pointers. However, such components are also often of unknown provenance and quality and may be riddled with accidental bugs or, in some cases, deliberately malicious code. Statistics show that such component failures account for a high percentage of software crashes and vulnerabilities. Enabling isolation of such fine-grained components is therefore necessary to increase the stability, security and resilience of computer programs. This thesis addresses this issue by showing how host applications can create isolation domains for individual components, while preserving the benefits of a single address space, via a new architecture for software isolation called LibVM. Towards this end, we define a specification which outlines the functional requirements for LibVM, identify the conditions under which these functional requirements can be met, define an abstract Application Programming Interface (API) that encompasses the general problem of isolating shared libraries, thus separating policy from mechanism, and prove its practicality with two concrete implementations based on hardware virtualization and system call interpositioning, respectively. The results demonstrate that hardware isolation minimises the difficulties encountered with software based approaches, while also reducing the size of the trusted computing base, thus increasing confidence in the solution’s correctness. This thesis concludes that, not only is it feasible to create such isolation domains for individual components, but that it should also be a fundamental operating system supported abstraction, which would lead to more stable and secure applications.