Evaluation of DNA enzymes targeted against the RNA component of human telomerase

Glioblastoma Multiforme (GBM) is a highly malignant form of brain cancer for which there is currently no effective cure. Consequently, developing new therapies and elucidating effective targets is crucial for this fatal disease. In recent years, DNA enzymes, deoxyribonucleic acid molecules with enzymatic activity, have emerged. In the same manner as ribozymes, DNA enzymes are able to effect cleavage of RNA in a sequence-specific manner, and operate with catalytic efficiency. In this study, two DNA enzymes were designed to target the template region of human telomerase RNA (hTR), utilising the 10-23 and 8-17 catalytic motifs elucidated by Santoro and Joyce (1997). Telomerase is an RNA-dependent DNA polymerase, which stabilises telomere lengths by adding hexameric repeats (TTAGGG in humans) to chromosome termini, thus preventing the telomere shortening that usually occurs during mitotic cell division. Telomerase activity, whilst absent in normal somatic tissues, is present in almost 90% of all tumours. Thus, there is speculation that telomerase may be the much sought universal target for therapeutic intervention in cancer. In vitro cleavage assays showed both DNA enzymes to be catalytically competent. Unmodified phosphodiester (PO) backbone DNA enzymes were rapidly degraded in the presence of serum, with a half-life of 10 minutes. The common approach of introducing phosphorothioate (PS) linkages was used in an effort to overcome this instability. As a result of concurrent activity and stability studies on the DNA enzymes with various numbers of PS linkages, the DNA enzymes with a PO core and PS arms were chosen for use in further cell work. The cleavage activity of both was shown to be specific and affected by temperature, pH, MgCI2 concentration and enzyme concentration. Both DNA enzyme motifs reduced telomerase activity in cell lysates, as assessed by the telomerase repeat amplification protocol (TRAP) with an IC50 of 100nM. DNA enzymes being polyanionic molecules do not readily cross biological barriers. Cellular association of naked DNA enzyme was inefficient at less than 2%. Cellular delivery of the DNA enzymes was effectively improved using commercial cationic lipid formulations. However, the lipid-mediated delivery of DNA enzymes to U87-MG cells over a 4-hour period did not significantly inhibit cell proliferation compared to controls. This is possibly due to an expected lag period between the inhibition of telomere maintenance and cell death. Therefore, biodegradable polymer microspheres were investigated as a potential delivery option for prolonged and sustained delivery. In vitro release profiles showed that after an initial burst, sustained release of DNA enzymes was observed over 35 days. Finally, the efficacy and specificity of the DNA enzymes were demonstrated in a luciferase based reporter assay. Specific inhibition of luciferase expression was displayed at 10nM. Thus DNA enzymes have potential against endogenous cellular targets.

Development and trialling of a behavioural intervention for patients with atrial fibrillation initiating oral anticoagulation:the 'treat' study

Purpose: Atrial fibrillation (AF) is the most common heart arrhythmia and is associated with an increased risk of stroke. Stroke risk is commonly treated with oral anticoagulation (OAC) with a narrow therapeutic range (INR 2.0 to 3.0); which is poorly controlled in practice. Barriers to adherence include poor knowledge, and inaccurate perceptions surrounding illness and medications. Trial registration: ISRCTN93952605. Systematic review: Seven trials of educational, self-monitoring and decision aid interventions were included in a systematic review. Pooled analysis suggested education OR, 95% CI 7.89 (5.54-10.24) and self monitoring OR (95% CI) 5.47(2.55-8.39) significantly improve TTR; whereas decision aids are no more effective in reducing decision conflict than usual care, OR (95% CI) -0.10 (-0.17 to -0.02). Intervention development: The intervention was theoretically-driven (utilising the common sense and beliefs about medication models) and developed with expert patient feedback. Described using behavioural change techniques, the one-off group session included an educational booklet, ‘expert-patient’ focussed DVD, and worksheet. Methods: Ninety seven warfarin-naïve AF patients were randomised to receive the intervention (n=43), or usual care (n=54). The primary endpoint was time within therapeutic range (TTR), secondary endpoints included knowledge, quality of life (AF-QoL-18), beliefs about medication (BMQ), illness perceptions (IPQ-B), and anxiety and depression (HADS). Results: Intervention group had significantly higher TTR than usual care (78.5% vs. 66.7%; p=0.01). Knowledge changed significantly across time (F (3, 47) = 6.4; p<0.01), but not between groups (F (1, 47) = 3.3; p = 0.07). At six months knowledge predicted TTR (r=0.245; p=0.04). Illness concern negatively correlated with TTR (r= - 0.199; p=0.05). General Harm scores at one month predicted TTR (F (1, 72) = 4.08; p=0.048). There were significant differences in emotional representations (F (3, 49) = 3.3 (3, 49); p= 0.03), anxiety (F (3, 46) = 25.2; p<0.01) and depression (F (3, 46) = 37.7; p<0.01) across time. Conclusion: A theory-driven educational intervention can improve TTR in AF patients and potentially reduce the risk of adverse clinical outcomes. Improving education provision for AF patients is essential to ensure efficacious treatment.

General introduction

Knowledge is of crucial, and growing importance in social, political and economic relations in modern society. The range and variety of available knowledge dramatically enlarges the available options of social action. Featuring essays from philosophers who have investigated the foundations of knowledge, and addressing different forms of knowledge in society such as common sense and practical knowledge, this collection also discusses the role of knowledge in economic process and gives attention to the role of expert knowledge in political decision making.

Knowledge:the foundations of knowledge

Knowledge is of crucial, and growing importance in social, political and economic relations in modern society. The range and variety of available knowledge dramatically enlarges the available options of social action. This five volume collection brings together a broad array of contributions from a variety of disciplines. Featuring essays from philosophers who have investigated the foundations of knowledge, and addressing different forms of knowledge in society such as common sense and practical knowledge, this collection also discusses the role of knowledge in economic process and gives attention to the role of expert knowledge in political decision making. Including a collection of articles from the sociology of knowledge and science, the set also provides a new introduction by the editors, making it a unique and invaluable research resource for both student and scholar.

Knowledge:politics and knowledge

Knowledge is of crucial, and growing importance in social, political and economic relations in modern society. The range and variety of available knowledge dramatically enlarges the available options of social action. This five volume collection brings together a broad array of contributions from a variety of disciplines. Featuring essays from philosophers who have investigated the foundations of knowledge, and addressing different forms of knowledge in society such as common sense and practical knowledge, this collection also discusses the role of knowledge in economic process and gives attention to the role of expert knowledge in political decision making. Including a collection of articles from the sociology of knowledge and science, the set also provides a new introduction by the editors, making it a unique and invaluable research resource for both student and scholar.

Knowledge

Knowledge is of crucial, and growing importance in social, political and economic relations in modern society. The range and variety of available knowledge dramatically enlarges the available options of social action. This five volume collection brings together a broad array of contributions from a variety of disciplines. Featuring essays from philosophers who have investigated the foundations of knowledge, and addressing different forms of knowledge in society such as common sense and practical knowledge, this collection also discusses the role of knowledge in economic process and gives attention to the role of expert knowledge in political decision making. Including a collection of articles from the sociology of knowledge and science, the set also provides a new introduction by the editors, making it a unique and invaluable research resource for both student and scholar.

Priority evaluation in transportation policies and programmes

This work is concerned with the development of techniques for the evaluation of large-scale highway schemes with particular reference to the assessment of their costs and benefits in the context of the current transport planning (T.P.P.) process. It has been carried out in close cooperation with West Midlands County Council, although its application and results are applicable elsewhere. The background to highway evaluation and its development in recent years has been described and the emergence of a number of deficiencies in current planning practise noted. One deficiency in particular stood out, that stemming from inadequate methods of scheme generation and the research has concentrated upon improving this stage of appraisal, to ensure that subsequent stages of design, assessment and implementation are based upon a consistent and responsive foundation. Deficiencies of scheme evaluation were found to stem from inadequate development of appraisal methodologies suffering from difficulties of valuation, measurement and aggregation of the disparate variables that characterise highway evaluation. A failure to respond to local policy priorities was also noted. A 'problem' rather than 'goals' based approach to scheme generation was taken, as it represented the current and foreseeable resource allocation context more realistically. A review of techniques with potential for highway problem based scheme generation, which would work within a series of practical and theoretical constraints were assessed and that of multivariate analysis, and classical factor analysis in particular, was selected, because it offerred considerable application to the difficulties of valuation, measurement and aggregation that existed. Computer programs were written to adapt classical factor analysis to the requirements of T.P.P. highway evaluation, using it to derive a limited number of factors which described the extensive quantity of highway problem data. From this, a series of composite problem scores for 1979 were derived for a case study area of south Birmingham, based upon the factorial solutions, and used to assess highway sites in terms of local policy issues. The methodology was assessed in the light of its ability to describe highway problems in both aggregate and disaggregate terms, to guide scheme design, coordinate with current scheme evaluation methods, and in general to improve upon current appraisal. Analysis of the results was both in subjective, 'common-sense' terms and using statistical methods to assess the changes in problem definition, distribution and priorities that emerged. Overall, the technique was found to improve upon current scheme generation methods in all respects and in particular in overcoming the problems of valuation, measurement and aggregation without recourse to unsubstantiated and questionable assumptions. A number of deficiencies which remained have been outlined and a series of research priorities described which need to be reviewed in the light of current and future evaluation needs.

“I thought I’d be safe there”:pre-empting blame in the talk of women reporting rape

One routine “common sense” means of explaining sexual violence is the ideologically facilitated tendency to blame the victim, and previous research has identified patterns of victim-blaming in the talk of perpetrators of rape, and also in that of the professionals who deal with rape in their day-to-day work. This article focuses on the discursive resources drawn on in police interviews by rape victims themselves as they attempt to account for their own behaviour in relation to the attack. It identifies and describes points within interviewees’ talk where they produce “accounts” (Potter and Wetherell, 1987), and considers what these tell us about the participants’ shared understanding of what is relevant to the on-going talk. Occasions when there is evidence of a mis-match in the understanding of the participants will also be discussed. The analyses illustrate that for the accounts of interviewees to be heard as relevant, a number of prevalent and problematic themes of victim-blaming must be assumed. Interviewees anticipate and pre-empt implications that various aspects of their own behaviour contributed to their attack, and interviewers vary in the level of skill they display at negotiating these shared understandings.

Developing a complex educational–behavioural intervention:the TREAT intervention for patients with atrial fibrillation

This article describes the theoretical and pragmatic development of a patient-centred intervention for patients with atrial fibrillation (AF). Theoretical models (Common Sense Model, Necessity-Concerns Framework), clinical frameworks, and AF patient feedback contributed to the design of a one-off hour-long behaviour-change intervention package. Intervention materials consisted of a DVD, educational booklet, diary and worksheet, which were patient-centred and easy to administer. The intervention was evaluated within a randomised controlled trial. Several “active theoretical ingredients” were identified (for e.g., where patients believed their medication was less harmful they spent more time within the therapeutic range (TTR), with general harm scores predicting TTR at 6 months). Allowing for social comparison and adopting behaviour change techniques enabled accurate patient understanding of their condition and medication. The process of developing the intervention using theory-derived content and evaluation tools allowed a greater understanding of the mechanisms by which this intervention was successful. Alleviating concerns about treatment medication by educating patients can help to improve adherence. This process of intervention development could be adopted for a range of chronic illnesses and treatments. Critical elements should include the use of: (1) clinical guidelines; (2) appropriate theoretical models; (3) patient input; and (4) appropriate evaluation tools.

Resisting the final word:challenging stale media and policy representations of students’ performative technological encounters in university education

This article explores powerful, constraining representations of encounters between digital technologies and the bodies of students and teachers, using corpus-based Critical Discourse Analysis (CDA). It discusses examples from a corpus of UK Higher Education (HE) policy documents, and considers how confronting such documents may strengthen arguments from educators against narrow representations of an automatically enhanced learning. Examples reveal that a promise of enhanced ‘student experience’ through information and communication technologies internalizes the ideological constructs of technology and policy makers, to reinforce a primary logic of exchange value. The identified dominant discursive patterns are closely linked to the Californian ideology. By exposing these texts, they provide a form of ‘linguistic resistance’ for educators to disrupt powerful processes that serve the interests of a neoliberal social imaginary. To mine this current crisis of education, the authors introduce productive links between a Networked Learning approach and a posthumanist perspective. The Networked Learning approach emphasises conscious choices between political alternatives, which in turn could help us reconsider ways we write about digital technologies in policy. Then, based on the works of Haraway, Hayles, and Wark, a posthumanist perspective places human digital learning encounters at the juncture of non-humans and politics. Connections between the Networked Learning approach and the posthumanist perspective are necessary in order to replace a discourse of (mis)representations with a more performative view towards the digital human body, which then becomes situated at the centre of teaching and learning. In practice, however, establishing these connections is much more complex than resorting to the typically straightforward common sense discourse encountered in the Critical Discourse Analysis, and this may yet limit practical applications of this research in policy making.

Measuring the impact of illness perception on NHS audiology service usage in presbycusis patients – a feasibility study

Objective: Coping behaviour in adult hearing loss is still not well understood. Despite the high prevalence of hearing loss in those over 65, many people do not seek help for hearing loss. The common sense model of illness perceptions suggests that illness perceptions are a strong predictor of adapted coping behaviours, including help-seeking and take-up of treatments. This study aimed to determine the feasibility of using the brief illness perceptions questionnaire (bIPQ) to measure the impact of illness perception in predicting usage of NHS audiology services. Study design: Twenty-four volunteers were recruited from a standard NHS audiology outpatient clinic and illness perception was measured using the bIPQ. Two different recruitment strategies were explored and compared in terms of recruitment and retention rates. Comprehensibility of the questionnaire was assessed by Think Aloud Analysis in a subset of participants, while possible risks and burdens were monitored in structured telephone interviews. Results: The questionnaire is a comprehensive and quick tool to measure individual illness perception at minimal cost. We suggested minor adaptations of three questionnaire items to increase comprehension. Participants preferred to complete the questionnaire after their appointment at the clinic facilities rather than at home prior to their hearing assessment appointment. There were no identified risks or burdens to participants in this study. Conclusions: This approach met our criteria for feasibility. Understanding the impact of illness perception on patients’ coping behaviour in presbycusis could improve treatment outcomes and increase patient satisfaction, while promoting a more efficient and individualized audiology service.

Whole life performance assessment: critical success factors

Whole life costing (WLC) has become the best practice in construction procurement and it is likely to be a major issue in predicting whole life costs of a construction project accurately. However, different expectations from different organizations throughout a project's life and the lack of data, monitoring targets, and long-term interest for many key players are obstacles to be overcome if WLC is to be implemented. A questionnaire survey was undertaken to investigate a set of ten common factors and 188 individual factors. These were grouped into eight critical categories (project scope, time, cost, quality, contract/administration, human resource, risk, and health and safety) by project phase, as perceived by the clients, contractors and subcontractors in order to identify critical success factors for whole life performance assessment (WLPA). Using a relative importance index, the top ten critical factors for each category, from the perspective of project participants, were analyzed and ranked. Their agreement on those categories and factors were analyzed using Spearman's rank correlation. All participants identify “Type of Project” as the most common critical factor in the eight categories for WLPA. Using the relative index ranking technique and weighted average methods, it was found that the most critical individual factors in each category were: “clarity of contract” (scope); “fixed construction period” (time); “precise project budget estimate” (cost); “material quality” (quality); “mutual/trusting relationships” (contract/administration); “leadership/team management” (human resource); and “management of work safety on site” (health and safety). There was relatively a high agreement on these categories among all participants. Obviously, with 80 critical factors of WLPA, there is a stronger positive relationship between client and contactor rather than contractor and subcontractor, client and subcontractor. Putting these critical factors into a criteria matrix can facilitate an initial framework of WLPA in order to aid decision making in the public sector in South Korea for evaluation/selection process of a construction project at the bid stage.

Learning for life:educational provision at Cadbury, 1831-1981

Cadbury showed concern for the welfare of its labour force in a variety of ways and not least in the provision of educational and educative-recreational facilities. The firm regarded the education of employees as having a positive effect on the efficiency of the business at the same time as being of benefit to the individual, the local community and the nation. The life-long education of people was seen as essential for personal fulfilament, social improvement, economic competitiveness and the proper functioning of democratic procedures. The educational system built up at Cadbury, and the philosophy on which it was founded, acquired both a domestic and international reputation. Its main components were the day continuation education of juniors; the Bournville Works Evening Institute; vocational and non-vocational scholarships; emphasis on the primary importance of general education as a basis for life, work and technical training; stress on equality of educational opportunity for females; and leisure and sporting amenities which the firm felt to be educative in the sense that they contributed to personal psychological and physical development and social skills. The system was primarily shaped and constructed in the first three decades of the twentieth century and went into decline and eventual demise in the 1960's and 1970's as a result of economic pressures, social changes, enhanced state arrangements for education, shifts in Cadbury management thinking and the merger with Schweppes in 1969.

Calcitonin and calcitonin receptor-like receptors:common themes with family B GPCRs?

The calcitonin receptor (CTR) and calcitonin receptor-like receptor (CLR) are two of the 15 human family B (or Secretin-like) GPCRs. CTR and CLR are of considerable biological interest as their pharmacology is moulded by interactions with receptor activity-modifying proteins. They also have therapeutic relevance for many conditions, such as osteoporosis, diabetes, obesity, lymphatic insufficiency, migraine and cardiovascular disease. In light of recent advances in understanding ligand docking and receptor activation in both the family as a whole and in CLR and CTR specifically, this review reflects how applicable general family B GPCR themes are to these two idiosyncratic receptors. We review the main functional domains of the receptors; the N-terminal extracellular domain, the juxtamembrane domain and ligand interface, the transmembrane domain and the intracellular C-terminal domain. Structural and functional findings from the CLR and CTR along with other family B GPCRs are critically appraised to gain insight into how these domains may function. The ability for CTR and CLR to interact with receptor activity-modifying proteins adds another level of sophistication to these receptor systems but means careful consideration is needed when trying to apply generic GPCR principles. This review encapsulates current thinking in the realm of family B GPCR research by highlighting both conflicting and recurring themes and how such findings relate to two unusual but important receptors, CTR and CLR.

The measurement properties of the menorrhagia multi-attribute quality-of-life scale:a psychometric analysis

Menorrhagia, or heavy menstrual bleeding (HMB), is a common gynaecological condition. As the aim of treatment is to improve women's wellbeing and quality of life (QoL), it is necessary to have effective ways to measure this. This study investigated the reliability and validity of the menorrhagia multi-attribute scale (MMAS), a menorrhagia-specific QoL instrument. Participants (n = 431) completed the MMAS and a battery of other tests as part of the baseline assessment of the ECLIPSE (Effectiveness and Cost-effectiveness of Levonorgestrel-containing Intrauterine system in Primary care against Standard trEatment for menorrhagia) trial. Analyses of their responses suggest that the MMAS has good measurement properties and is therefore an appropriate condition-specific instrument to measure the outcome of treatment for HMB. © 2011 The Authors BJOG An International Journal of Obstetrics and Gynaecology © 2011 RCOG.