An investigation of partnership working in the implementation and delivery of the 14-19 diploma

D. EdIn this thesis I investigated partnership working in the development and implementation of the 14-19 diploma in the North-East of England. The research encompassed the period from the implementation of the 14-19 diploma in 2008 to its withdrawal in 2013 by the Conservative-Liberal coalition government. The focus of my research was whether partnership was happening and how partnership working was viewed and undertaken by those involved. I used a mixed-methods approach to examine data from participants across North-East England, including diploma practitioners, learners, and parents to gain their perspective on diploma design and implementation. My methodology comprised electronic questionnaires, semi-structured interviews, and a personal notebook supported by secondary research. I collected and analysed qualitative and quantitative data to investigate my two research questions: Was partnership working taking place in the development and delivery of the diploma in the North-East of England? What did ‘partnership’ mean to the stakeholders involved in the development and delivery of the diploma? I analysed the data in relation to these questions which allowed me to identify and discuss further themes from within the responses, including; competition, collaboration, and the specific nature of partnership working in education. I then considered these themes in terms of their effect on partnership working and how they related to theory explored in the literature review. From the data analysis I identified a new concept in relation to partnership working in education. I have contributed to the understanding of the development and implementation of the diploma in the region, to the understanding of partnership working in 14-19 education, and to wider partnership working knowledge. The results were particularly useful for exploring issues of competition and collaboration, and understanding how partnership work was viewed and used by practitioners, learners and parents involved with the 14-19 diploma

Urban society and the English Revolution : the archaeology of the new Jerusalem

PhD ThesisThe English Revolution has long been a defining subject of English historiography, with a large and varied literature that reflects continuing engagement with the central themes of civil conflict, and deep-rooted social, political and religious change. By contrast, this period has failed to catch the imagination of archaeologists. This research seeks to understand the world of the English Revolution through its material expression in English towns. Identifying the material expressions of the period is central to developing an archaeological understanding of the period. The clearest material expressions are found, in the fortifications that were built to protect towns, the destruction that was wrought on towns and in the reconstruction of the material world of English towns. Towns, like any other artefact, have their meanings. These meanings are multivalent and ever shifting, defined by the interaction of their material fabric and those who experience it. As these meanings change over time, they can be traced through the structures and artefacts of the town, and through the myths and legends that accrete on them. Understanding the interactions of material, myth and memory allows archaeologists to understand the true meaning of the urban built environment to generate a deeper and more nuanced understanding of the nature of the English urban culture of the period. Towns were fundamental to the English imagination as much as they were economically, politically or socially important. The English Revolution sits at the heart of the accepted conception of historical archaeology, but has been curiously neglected by historical archaeologists. The cultural conflict of this period embodies the themes that are central to historical archaeology, and nowhere is this more apparent than in urban culture

The contribution of plant cells and their enzymes to the biochemical changes which take place in the ensilage process

A growth-chamber is described in which attempts have been made to grow microbe-free timothy grass from which silage can be made without chance of contamination. It was expected that useful information about the chemical changes due to plant cells in the ensilage process would result from analyses of the products.The attempts were only partially successful; sterile silage was produced in one case only but some tentative conclusions could be drawn from the results of other experiments in which only a very restricted microflora was presentAs a result of plant cell metabolism the protein breakdown, production of volatile base and possibly of alcohol occurred to an extent comparable to that observed in normal silage. No significant increase in lactic or volatile acids took place but a definite increase in succinic acid was noted although no quantitative data were obtained. Hydrogen, which is normally evolved by grass silage in tubes, does not arise as a result of plant cell activity.New methods are presented for the quantitative estimation of volatile base and alcohol in silage and qualitative methods of analysis are described for amino-acids, volatile acids and other carboxylic acids.It is considered that the growth-chamber can be further improved in design and with the technique described should prove of use in other similar studies to those reported and also in other plant physiological investigations

Liquid Hydrogen Pressurization Tests Final Test Report, 29 May 1963 - 27 Aug. 1964

Liquid hydrogen pressurization tests for application to heat exchanger design in Saturn missil

Laboratory-directed evolution as a tool for anticipating insecticide resistance

The evolution of insecticide resistance provides a eukaryotic model system for studying enzyme evolution. Understanding the molecular basis of insecticide resistance can assist both the development of new methods to combat resistance and the anticipation of future resistance. Three insect species have independently evolved catalytic organophosphate (OP) insecticide resistance through a single active-site mutation (Gly\u3eAsp) in the αE7 enzyme1-3. To explore the evolutionary potential of αE7, we subjected αE7 from the blowfly Lucilia cuprina to nine rounds of mutation and selection, resulting in a \u3e1000-fold increase in OP-hydrolase activity and a kcat / KM \u3e 106 M-1 min-1. Kinetic and structural analysis of the evolutionary trajectory revealed the molecular basis for the increase in catalytic efficiency. Mutations occurring in the early stages of the trajectory enrich the productive side chain conformation of the key aspartic acid residue, while mutations in later stages remodel the binding pocket. Remarkably, mutations appearing in the later rounds yielded larger improvements in catalytic efficiency compared to initial mutations, indicating that the initial Gly\u3eAsp mutation represents only a fraction of the αE7 evolutionary potential. Worryingly, this suggests that the Gly\u3eAsp could be the first of many steps toward efficient OP-insecticide detoxification. Please click Additional Files below to see the full abstract

The evolution of multiple active site configurations in a designed enzyme

Developments in computational chemistry, bioinformatics, and laboratory evolution have facilitated the de novo design and catalytic optimization of enzymes. Besides creating useful catalysts, the generation and iterative improvement of designed enzymes can provide valuable insight into the interplay between the many phenomena that have been suggested to contribute to catalysis. In this work, we follow changes in conformational sampling, electrostatic preorganization, and quantum tunneling along the evolutionary trajectory of a designed Kemp eliminase. We observe that in the Kemp Eliminase KE07, instability of the designed active site leads to the emergence of two additional active site configurations. Evolutionary conformational selection then gradually stabilizes the most efficient configuration, leading to an improved enzyme. This work exemplifies the link between conformational plasticity and evolvability and demonstrates that residues remote from the active sites of enzymes play crucial roles in controlling and shaping the active site for efficient catalysis

Activity-based E3 ligase profiling uncovers an E3 ligase with esterification activity

Ubiquitination is initiated by transfer of ubiquitin (Ub) from a ubiquitin-activating enzyme (E1) to a ubiquitin-conjugating enzyme (E2), producing a covalently linked intermediate (E2-Ub)(1). Ubiquitin ligases (E3s) of the 'really interesting new gene' (RING) class recruit E2-Ub via their RING domain and then mediate direct transfer of ubiquitin to substrates(2). By contrast, 'homologous to E6-AP carboxy terminus' (HECT) E3 ligases undergo a catalytic cysteine-dependent transthiolation reaction with E2-Ub, forming a covalent E3-Ub intermediate(3,4). Additionally, RING-between-RING (RBR) E3 ligases have a canonical RING domain that is linked to an ancillary domain. This ancillary domain contains a catalytic cysteine that enables a hybrid RING-HECT mechanism(5). Ubiquitination is typically considered a post-translational modification of lysine residues, as there are no known human E3 ligases with non-lysine activity. Here we perform activity-based protein profiling of HECT or RBR-like E3 ligases and identify the neuron-associated E3 ligase MYCBP2 (also known as PHR1) as the apparent single member of a class of RING-linked E3 ligase with esterification activity and intrinsic selectivity for threonine over serine. MYCBP2 contains two essential catalytic cysteine residues that relay ubiquitin to its substrate via thioester intermediates. Crystallographic characterization of this class of E3 ligase, which we designate RING-Cys-relay (RCR), provides insights into its mechanism and threonine selectivity. These findings implicate non-lysine ubiquitination in cellular regulation of higher eukaryotes and suggest that E3 enzymes have an unappreciated mechanistic diversity

An Atypical E3 Ligase Module in UBR4 Mediates Destabilization of N-degron Substrates

UBR4 is an E3 ligase (E3) of the N-degron pathway and is involved in neurodevelopment, age-associated muscular atrophy and cancer progression. The location and mechanistic classification of the E3 module within the 600 kDa protein UBR4 remains unknown. Herein, we identify and characterize, at a biochemical and structural level, a distinct E3 module within human UBR4 consisting of a novel “hemiRING” zinc finger, a helical-rich UBR Zinc-finger Interacting (UZI) subdomain, and a predicted backside interacting N-terminal helix. A structure of an E2 conjugating enzyme (E2)-E3 complex provides atomic level insight into the exquisite specificity of the hemiRING towards the E2s UBE2A/B. The UZI subdomain can be considered a component of the E3 module as it has a modest activating effect on the ubiquitin loaded E2 (E2∼Ub), which is complemented by the intrinsically high lysine reactivity of UBE2A. These findings reveal the mechanistic underpinnings of a neuronal N-degron E3 ligase, its specific recruitment of UBE2A, and highlight the underappreciated architectural diversity of cross-brace domains associated with ubiquitin E3 activity.<br/