The redoxomics of PTEN: walking a fine line between damage and signaling:mass spectrometry-based approaches to study the effect of oxidation on PTEN function, structure, and protein-protein interactions

The research described in this PhD thesis focuses on proteomics approaches to study the effect of oxidation on the modification status and protein-protein interactions of PTEN, a redox-sensitive phosphatase involved in a number of cellular processes including metabolism, apoptosis, cell proliferation, and survival. While direct evidence of a redox regulation of PTEN and its downstream signaling has been reported, the effect of cellular oxidative stress or direct PTEN oxidation on PTEN structure and interactome is still poorly defined. In a first study, GST-tagged PTEN was directly oxidized over a range of hypochlorous acid (HOCl) concentration, assayed for phosphatase activity, and oxidative post-translational modifications (oxPTMs) were quantified using LC-MS/MS-based label-free methods. In a second study, GSTtagged PTEN was prepared in a reduced and reversibly H2O2-oxidized form, immobilized on a resin support and incubated with HCT116 cell lysate to capture PTEN interacting proteins, which were analyzed by LC-MS/MS and comparatively quantified using label-free methods. In parallel experiments, HCT116 cells transfected with a GFP-tagged PTEN were treated with H2O2 and PTENinteracting proteins immunoprecipitated using standard methods. Several high abundance HOCl-induced oxPTMs were mapped, including those taking place at amino acids known to be important for PTEN phosphatase activity and protein-protein interactions, such as Met35, Tyr155, Tyr240 and Tyr315. A PTEN redox interactome was also characterized, which identified a number of PTEN-interacting proteins that vary with the reversible inactivation of PTEN caused by H2O2 oxidation. These included new PTEN interactors as well as the redox proteins peroxiredoxin-1 (Prdx1) and thioredoxin (Trx), which are known to be involved in the recycling of PTEN active site following H2O2-induced reversible inactivation. The results suggest that the oxidative modification of PTEN causes functional alterations in PTEN structure and interactome, with fundamental implications for the PTEN signaling role in many cellular processes, such as those involved in the pathophysiology of disease and ageing.

Brain oscillations and connectivity in autism spectrum disorders (ASD):new approaches to methodology, measurement and modelling

Although atypical social behaviour remains a key characterisation of ASD, the presence ofsensory and perceptual abnormalities has been given a more central role in recentclassification changes. An understanding of the origins of such aberrations could thus prove afruitful focus for ASD research. Early neurocognitive models of ASD suggested that thestudy of high frequency activity in the brain as a measure of cortical connectivity mightprovide the key to understanding the neural correlates of sensory and perceptual deviations inASD. As our review shows, the findings from subsequent research have been inconsistent,with a lack of agreement about the nature of any high frequency disturbances in ASD brains.Based on the application of new techniques using more sophisticated measures of brainsynchronisation, direction of information flow, and invoking the coupling between high andlow frequency bands, we propose a framework which could reconcile apparently conflictingfindings in this area and would be consistent both with emerging neurocognitive models ofautism and with the heterogeneity of the condition.

A novel approach to modelling and exploiting uncertainty in stochastic control systems

We consider an inversion-based neurocontroller for solving control problems of uncertain nonlinear systems. Classical approaches do not use uncertainty information in the neural network models. In this paper we show how we can exploit knowledge of this uncertainty to our advantage by developing a novel robust inverse control method. Simulations on a nonlinear uncertain second order system illustrate the approach.

Positive psychology approaches to public services leadership: An introduction to strengths-based leadership

To readers of the popular press, the words ‘positive psychology’ may conjure up images of happiness gurus and people having their feet massaged, their heads resting peacefully on pink, fluffy clouds. But in this article, our aim is to demonstrate how the new science of positive psychology speaks powerfully to - and has much to contribute to - the development of leadership and the practices and processes of organisations, whether in the public or private sectors. Much of our work is concerned with the applications of this new field, and particularly with building strengths-based organisations. A key pillar of this work is around enabling strengths-based leadership, and provides our focus for this article.

Evolving approaches to sovereignty in the French Pacific

This contribution explores notions of sovereignty in the three French territories of the South Pacific: French Polynesia, New Caledonia, and Wallis and Futuna. It also analyses the key nuances and challenges of the transition from aspirations of 'independence' to those of 'shared sovereignty'. From protectorates or colonies to overseas territories (Territoires d'Outre-Mer), these three territories have experienced specific and customised statuses with various degrees of autonomy, all underscoring a fine line between autonomy and sovereignty. Indeed, 'sovereignty' has today become much more synonymous with the concept of 'self-government' or 'large autonomy', as the current situation in French Polynesia demonstrates. Meanwhile, New Caledonia is one step closer to 'full sovereignty', since its actual status includes provisions for a referendum on self-determination between 2014 and 2018. The claim for independence, a characteristic of Kanak and Maohi movements, has become more pragmatically focused, to the extent that considering sovereignty 'in free-association' with France is now a perfectly conceivable option. © 2012 Copyright Taylor and Francis Group, LLC.

Approaches to sharing knowledge in group problem structuring

Problem-structuring techniques are an integral aspect of 'Soft-OR'. SSM, SAST, Strategic Choice, and JOURNEY Making, all depend for their success on a group developing a shared view of a problem through some form of explicit modelling. The negotiated problem structure becomes the basis for problem resolution. Implicit to this process is an assumption that members of the group share and build their knowledge about the problem domain. This paper explores the extent to which this assumption is reasonable. The research is based on detailed records from the use of JOURNEY Making, where it has used special purpose Group Support software to aid the group problem structuring. This software continuously tracks the contributions of each member of the group and thus the extent to which they appear to be 'connecting' and augmenting their own knowledge with that of other members of the group. Software records of problem resolution in real organisational settings are used to explore the sharing of knowledge among senior managers. These explorations suggest a typology of knowledge sharing. The implications of this typology for problem structuring and an agenda for future research are considered.

Approaches to issues of job regulation. A study at branch level in a white collar, public sector trade union. Available in 2 volumes.

This study examines the internal dynamics of white collar trade union branches in the public sector. The effects of a number of internal and external factors on branch patterns of action are evaluated. For the purposes of the study branch action is taken to be the approach to issues of job regulation, as expressed along the five dimensions of dependence on the outside trade union, focus in issues adopted, initiation of issues, intensity of action in issue pursuit and representativeness. The setting chosen for the study is four branches drawn from the same geographical area of the National and Local Government Officers Association. Branches were selected to give a variety in industry settings while controlling for the potentially influential variables of branch size, density of trade union membership and possession of exclusive representational rights in the employing organisation. Identical methods of data collection were used for each branch. The principal findings of the study are that the framework of national agreements and industry collective bargaining structures are strongly related to the industrial relations climate in the employing organisation and the structures of representation within the branch. Where agreements and collective bargaining structures formally restrict branch job regulation roles, there is a degree of devolution of bargaining authority from branch level negotiators to autonomous shop stewards at workplace level. In these circumstances industrial relations climate is characterised by a degree of informality in relationships between management and trade union activists. In turn, industrial relations climate and representative structures together with actor attitudes, have strong effects on all dimensions of approach to issues of job regulation.

The investigation of vegetation change using remote sensing to detect and monitor migration of landfill gas

Decomposition of domestic wastes in an anaerobic environment results in the production of landfill gas. Public concern about landfill disposal and particularly the production of landfill gas has been heightened over the past decade. This has been due in large to the increased quantities of gas being generated as a result of modern disposal techniques, and also to their increasing effect on modern urban developments. In order to avert diasters, effective means of preventing gas migration are required. This, in turn requires accurate detection and monitoring of gas in the subsurface. Point sampling techniques have many drawbacks, and accurate measurement of gas is difficult. Some of the disadvantages of these techniques could be overcome by assessing the impact of gas on biological systems. This research explores the effects of landfill gas on plants, and hence on the spectral response of vegetation canopies. Examination of the landfill gas/vegetation relationship is covered, both by review of the literature and statistical analysis of field data. The work showed that, although vegetation health was related to landfill gas, it was not possible to define a simple correlation. In the landfill environment, contribution from other variables, such as soil characteristics, frequently confused the relationship. Two sites are investigated in detail, the sites contrasting in terms of the data available, site conditions, and the degree of damage to vegetation. Gas migration at the Panshanger site was dominantly upwards, affecting crops being grown on the landfill cap. The injury was expressed as an overall decline in plant health. Discriminant analysis was used to account for the variations in plant health, and hence the differences in spectral response of the crop canopy, using a combination of soil and gas variables. Damage to both woodland and crops at the Ware site was severe, and could be easily related to the presence of gas. Air photographs, aerial video, and airborne thematic mapper data were used to identify damage to vegetation, and relate this to soil type. The utility of different sensors for this type of application is assessed, and possible improvements that could lead to more widespread use are identified. The situations in which remote sensing data could be combined with ground survey are identified. In addition, a possible methodology for integrating the two approaches is suggested.

Protein oxidative modifications:new mass spectrometry approaches to detection in biological samples

In inflammatory diseases, release of oxidants leads to oxidative damage to proteins. The precise nature of oxidative damage to individual proteins depends on the oxidant involved. Chlorination and nitration are markers of modification by the myeloperoxidase-H2O2-Cl- system and nitric oxide-derived oxidants, respectively. Although these modifications can be detected by western blotting, currently no reliable method exists to identify the specific sites damage to individual proteins in complex mixtures such as clinical samples. We are developing novel LCMS2 and precursor ion scanning methods to address this. LC-MS2 allows separation of peptides and detection of mass changes in oxidized residues on fragmentation of the peptides. We have identified indicative fragment ions for chlorotyrosine, nitrotyrosine, hydroxytyrosine and hydroxytryptophan. A nano-LC/MS3 method involving the dissociation of immonium ions to give specific fragments for the oxidized residues has been developed to overcome the problem of false positives from ions isobaric to these immonium ions that exist in unmodified peptides. The approach has proved able to identify precise protein modifications in individual proteins and mixtures of proteins. An alternative methodology involves multiple reaction monitoring for precursors and fragment ions are specific to oxidized and chlorinated proteins, and this has been tested with human serum albumin. Our ultimate aim is to apply this methodology to the detection of oxidative post-translational modifications in clinical samples for disease diagnosis, monitoring the outcomes of therapy, and improved understanding of disease biochemistry.

Rewarding customer service? Using reward and recognition to deliver your customer service strategy

This report examines the role of reward and recognition in helping organisations deliver excellent customer service. It identifies the impact that reward and recognition practice has upon customer service and highlights which practices are more effective; shows the extent to which employees are satisfied with the rewards and recognition they receive; indicates those approaches to reward and recognition most associated with staff commitment; and looks at how other factors affect employee satisfaction with reward and recognition.

Social cognition in leader–follower relationship:applying insights from relationship science to understanding relationship-based approaches to leadership

Relationship-based approaches to leadership (e.g., Leader–Member Exchange theory) currently represent one of the most popular approaches to understanding workplace leadership. Although the concept of “relationship” is central to these approaches, generally this has not been well articulated and is often conceptualized simply in terms of relationship quality between the leader and the follower. In contrast, research in the wider relationship science domain provides a more detailed exposition of relationships and how they form and develop. We propose that research and methodology developed in relationship science (i.e., close relationships) can enhance understanding of the leader–follower relationship and therefore advance theory in this area. To address this issue, we organize our review in two areas. First, we examine how a social cognitive approach to close relationships can benefit an understanding of the leader–follower relationship (in terms of structure, content, and processes). Second, we show how the research designs and methodologies that have been developed in relationship science can be applied to understand better the leader–follower relationship. The cross-fertilization of research from the close relationships literature to understanding the leader–follower relationship provides new insights into leadership processes and potential avenues for further research. Copyright © 2013 John Wiley & Sons, Ltd.

Developing critical understanding in HRM students:using innovative teaching methods to encourage deep approaches to study

Purpose – This paper aims to focus on developing critical understanding in human resource management (HRM) students in Aston Business School, UK. The paper reveals that innovative teaching methods encourage deep approaches to study, an indicator of students reaching their own understanding of material and ideas. This improves student employability and satisfies employer need. Design/methodology/approach – Student response to two second year business modules, matched for high student approval rating, was collected through focus group discussion. One module was taught using EBL and the story method, whilst the other used traditional teaching methods. Transcripts were analysed and compared using the structure of the ASSIST measure. Findings – Critical understanding and transformative learning can be developed through the innovative teaching methods of enquiry-based learning (EBL) and the story method. Research limitations/implications – The limitation is that this is a single case study comparing and contrasting two business modules. The implication is that the study should be replicated and developed in different learning settings, so that there are multiple data sets to confirm the research finding. Practical implications – Future curriculum development, especially in terms of HE, still needs to encourage students and lecturers to understand more about the nature of knowledge and how to learn. The application of EBL and the story method is described in a module case study – “Strategy for Future Leaders”. Originality/value – This is a systematic and comparative study to improve understanding of how students and lecturers learn and of the context in which the learning takes place.