Novel Pentameric Structure of the Diarrhea-Inducing Region of the Rotavirus Enterotoxigenic Protein NSP4

A novel pentameric structure which differs from the previously reported tetrameric form of the diarrhea-inducing region of the rotavirus enterotoxin NSP4 is reported here. A significant feature of this pentameric form is the absence of the calcium ion located in the core region of the tetrameric structures. The lysis of cells, the crystallization of the region spanning residues 95 to 146 of NSP4 (NSP4(95-146)) of strain ST3 (ST3: NSP4(95-146)) at acidic pH, and comparative studies of the recombinant purified peptide under different conditions by size-exclusion chromatography (SEC) and of the crystal structures suggested pH-, Ca(2+)-, and protein concentration-dependent oligomeric transitions in the peptide. Since the NSP4(95-146) mutant lacks the N-terminal amphipathic domain (AD) and most of the C-terminal flexible region (FR), to demonstrate that the pentameric transition is not a consequence of the lack of the N- and C-terminal regions, glutaraldehyde cross-linking of the Delta N72 and Delta N94 mutant proteins, which contain or lack the AD, respectively, but possess the complete C-terminal FR, was carried out. The results indicate the presence of pentamers in preparations of these longer mutants. Detailed SEC analyses of Delta N94 prepared under different conditions, however, revealed protein concentration-dependent but metal ion-and pH-independent pentamer accumulation at high concentrations which dissociated into tetramers and lower oligomers at low protein concentrations. While calcium appeared to stabilize the tetramer, magnesium in particular stabilized the dimer. Delta N72 existed primarily in the multimeric form under all conditions. These findings of a calcium-free NSP4 pentamer and its concentration-dependent and largely calcium-independent oligomeric transitions open up a new dimension in an understanding of the structural basis of its multitude of functions.

A new pentameric structure of rotavirus NSP4 revealed by molecular replacement

The region spanning residues 95-146 of the rotavirus nonstructural protein NSP4 from the asymptomatic human strain ST3 has been purified and crystallized and diffraction data have been collected to a resolution of 2.6 angstrom. Several attempts to solve the structure by the molecular-replacement method using the available tetrameric structures of this domain were unsuccessful despite a sequence identity of 73% to the already known structures. A more systematic approach with a dimer as the search model led to an unexpected pentameric structure using the program Phaser. The various steps involved in arriving at this molecular-replacement solution, which unravelled a case of subtle variation between different oligomeric states unknown at the time of solving the structure, are presented in this paper.

Severe diffraction anisotropy, rotational pseudosymmetry and twinning complicate the refinement of a pentameric coiled-coil structure of NSP4 of rotavirus

The crystal structure of the region spanning residues 95-146 of the rotavirus nonstructural protein NSP4 from the asymptomatic human strain ST3 was determined at a resolution of 2.5 angstrom. Severe diffraction anisotropy, rotational pseudo-symmetry and twinning complicated the refinement of this structure. A systematic explanation confirming the crystal pathologies and describing how the structure was successfully refined is given in this report.

La crisis de 1930 y las políticas del New Deal : un examen desde la economía y las instituciones

Resumen: La presente investigación consiste en un análisis, desde la perspectiva de la economía y las instituciones, de las políticas del New Deal, implementado por Franklin D. Roosevelt en los Estados Unidos durante la crisis económica de los años 30. Para realizar dicho análisis se presentan los elementos principales de la crisis de 1930, luego se realiza un estudio de las políticas económicas aplicadas en el contexto de un marco de gran innovación institucional, y concluye evaluando las políticas expuestas, tanto desde el enfoque de sus resultados macroeconómicos como de su desempeño institucional. De esta manera el presente documento incluye un diagnóstico histórico del contexto norteamericano en aquel entonces y resalta los aspectos que motivaron a dicha administración a la aplicación de New Deal. Finalmente, se realiza un análisis del impacto macroeconómico de las políticas aplicadas. La conclusión sintetiza las principales enseñanzas de la experiencia desde la perspectiva de política económica y del diseño de la regulación, para concluir con algunas reflexiones acerca de su posible implementación en la actualidad.

Obasan, by Joy Kogawa, and Alias Grace, by Margaret Atwood: fictional representations of Canadian identity and History

Esta dissertação investiga de que maneiras a representação do sujeito canadense pode ser encontrada em dois romances representativos da literatura canadense contemporânea: Obasan, de Joy Kogawa, e Alias Grace, de Margaret Atwood. Esta investigação também demonstra que a busca pela definição da identidade canadense tem sido tema constante e relevante da cultura deste país. A indefinição quanto ao que significa ser canadense também tem permeado a literatura canadense ao longo dos séculos, notadamente desde o século XIX. A fim de observar a representação literária da busca pela definição da identidade canadense, esta investigação aborda os conceitos relativos à representação de grupos subalternos tradicionalmente silenciados. A análise comparativa dos romances citados contempla a relação entre memória e trauma autobiográficos, assim como as semelhanças narrativas entre ficção e história. Esta investigação também verifica de que maneiras a literatura pós-moderna emprega documentação oficial, relatos históricos e dados (auto) biográficos a serviço da reescrita da história através da metaficção historiográfica

News from Academy Bay

A spate of Galapagos books. The phenomenal El Nino. The Galapagos on television. Galapagos tourism. The CDRS Director in the Soviet Union. Saving the Hawaiian Petrel. Wild dogs and land iguanas. Franklin Delano Roosevelt and the Galapagos Islands. More about those bloodthirsty "vampire finches". Auf Wiedersehen, Friedemann! Harold Jefferson Coolidge at eighty. Events and visits at the Darwin Research Station.

An assessment of scup (Stenotomus chrysops) and black sea bass (Centropristas striata) discards in the directed otter trawl fisheries in the Mid-Atlantic Bight

This study was undertaken to re-assess the level of scup (Stenotomus chrysops) discards by weight and to evaluate the effect of various codend mesh sizes on the level of scup discards in the winter-trawl scup fishery. Scup discards were high in directed scup tows regardless of codend mesh — typically one to five times the weight of landings. The weight of scup discards in the present study did not differ significantly from that recorded in scup-targeted tows in the NMFS observer database. Most discards were required as such by the 22.86 cm TL (total length) fish-size limit for catches. Mesh sizes ≤12.7 cm, including the current legal mesh size (11.43 cm) did not adequately filter out scup smaller than 22.86 cm. The median length of scup discards was about 19.83 cm TL. Lowering the legal size for scup from 22.86 to 19.83 cm TL would greatly reduce discard mortality. Scup discards were a small fraction (0.4%) of black sea bass (Centropristis striata) landings in blacksea-bass−targeted tows. The black sea bass fishery is currently regulated under the small-mesh fishery gearrestricted area plan in which fishing is prohibited in some areas to reduce scup mortality. Our study found no evidence to support the efficacy of this management approach. The expectations that discarding would increase disproportionately as the trip limit (limit [in kilograms] on catch for a species) was reached towards the end of the trip and that discards would increase when the trip limit was reduced from 4536 kg to 454 kg at the end of the directed fishing season were not supported. Trip limits did not significantly affect discard mortality.

Bottom simulating reflector and gas seepage in Okinawa Trough: Evidence of gas hydrate in an active back-arc basin

To look for gas hydrate, 22 multi-channel and 3 single-channel seismic lines on the East China Sea (ECS) shelf slope and at the bottom of the Okinawa Trough were examined. It was found that there was indeed bottom simulating reflector (BSR) occurrence, but it is very rare. Besides several BSRs, a gas seepage was also found. As shown by the data, both the BSR and gas seepage are all related with local geological structures, such as mud diapir, anticline, and fault-controlled graben-like structure. However, similar structural "anomalies" are quite common in the tectonically very active Okinawa Trough region, but very few of them have developed BSR or gas seepage. The article points out that the main reason is probably the low concentration of organic carbon of the sediment in this area. It was speculated that the rare occurrence of gas hydrates in this region is governed by structure-controlled fluid flow. Numerous faults and fractures form a network of high-permeability channels in the sediment and highly fractured igneous basement to allow fluid circulation and ventilation. Fluid flow in this tectonic environment is driven primarily by thermal buoyancy and takes place on a wide range of spatial scales. The fluid flow may play two roles to facilitate hydrate formation: to help gather enough methane into a small area and to modulate the thermal regime.

Becoming a CEO: an exploration of the theory and practice of effective organisational leadership

This Thesis is an exploration of potential enhancement in effectiveness, personally, professionally and organisationally through the use of Theory as an Apparatus of Thought. Enhanced effectiveness was sought by the practitioner (Subject), while in transition to becoming Chief Executive of his organization. The introduction outlines the content and the structure of the University College Cork DBA. Essay One outlines what Theory is, what Adult Mental Development is and an exploration of Theories held in the Authors past professional practice. Immunity to change is also reflected on. Essay Two looks at the construct of the key Theories used in the Thesis. Prof. Robert Kegan’s Theory of Adult Mental Development was used to aid the generation of insight. The other key Theories used were The Theory of The Business, Theory of the Co‐operative and a Theory of Organisational Leadership. Essay Three explores the application of the key Theories in a professional setting. The findings of the Thesis were that the subject was capable of dealing with increased environmental complexity and uncertainty by using Theory as an Apparatus of Thought, which in turn enhanced personal, professional and organisational effectiveness. This was achieved by becoming more aware of the Theories held by the practitioner, the experiences from the application of those Theories, which then led to greater insight. The author also found that a detailed understanding of the Theory of the Business and a Theory of Leadership would support any new CEO in the challenging early part of their tenure.

Reasoning about trust, entrepreneurial services, and the virtual team

This Portfolio of Exploration (PoE) tracks a transformative learning developmental journey that is directed at changing meaning making structures and mental models within an innovation practice. The explicit purpose of the Portfolio is to develop new and different perspectives that enable the handling of new and more complex phenomena through self transformation and increased emotional intelligence development. The Portfolio provides a response to the question: ‘What are the key determinants that enable a Virtual Team (VT) to flourish where flourishing means developing and delivering on the firm’s innovative imperatives?’ Furthermore, the PoE is structured as an investigation into how higher order meaning making promotes ‘entrepreneurial services’ within an intra-firm virtual team, with a secondary aim to identify how reasoning about trust influence KGPs to exchange knowledge. I have developed a framework which specifically focuses on the effectiveness of any firms’ Virtual Team (VT) through transforming the meaning making of the VT participants. I hypothesized it is the way KGPs make meaning (reasoning about trust) which differentiates the firm as a growing firm in the sense of Penrosean resources: ‘inducement to expand and a limit of expansion’ (1959). Reasoning about trust is used as a higher order meaning-making concept in line with Kegan’s (1994) conception of complex meaning making, which is the combining of ideas and data in ways that transform meaning and implicates participants to find new ways of knowledge generation. Simply, it is the VT participants who develop higher order meaning making that hold the capabilities to transform the firm from within, providing a unique competitive advantage that enables the firm to grow.

Explorations in a meaning making system: Kerry as an entrepreneurial platform and the equine sector as a cluster

This Portfolio is about the changes that can be supported and achieved through transformational education that impacts on personal, professional and organisational levels. Having lived through an era of tremendous change over the second half of the twentieth century and into the twenty-first the author has a great drawing board to contemplate in the context of professional career experience as an engineer. The ability to engage in ‘subject-object’ separation is the means by which Kegan (1994, 2009) explains that transformation takes place and the Essays in this Portfolio aim to support and bring about such change. Exploration of aspects of ‘Kerry’ is the material selected to both challenge support change in the way of knowing from being subject to certain information and knowledge that to being able to consider it more objectively. The task of being able to distance judgement about the economy and economic development of Kerry was facilitated by various readings around of a number of key thinkers including Kegan, Drucker, Porter and Penrose. The central themes of Kerry or the potential for economic development are built into each Essay. Essay One focuses on reflections of Kerry life - on Kerry people within and without Kerry - and events as they affected understandings of how people related to and worked with one another. These reflections formed the basis for transformational goals identified which required a shift from an engineering mindset to encompass an economics-based view. In Essay Two knowledge of economic concepts is developed by exploring the writings of Drucker, Penrose, and Porter with pertinence to considering economic development generally, and for Kerry in particular in the form of an ‘entrepreneurial platform’. The concepts and theories were the basis of explorations presented in Essays Three and Four. Essay Three focuses on Kerry’s potential for economic development give its current economic profile and includes results from interviews with selected businesses. Essay Four is an exercise in the application of Porter’s ‘Cluster’ concept to the equine sector.

Inquiring into perspective transformation in university education

My Portfolio of Exploration tackles the difficult question as to whether adult mental development can be accelerated and if so how. Rooted in constructive-developmental ideas, adult mental development is explained as an evolutionary unfolding of human capability. Going beyond this I look at the possibility of advancing development as transformational growth in adulthood in the belief that a broader perspective leads to increased effectiveness in professional life. Initially I explored my own meaning making, to make sense of my experiences, knowledge, relationships and my own motivations. This exploration has provided me with a ‘developmental bridge’ between my current way of knowing and a new more enlightened way. I have come to view my way of making meaning in the world as an evolving and progressive sequence of emotional and cognitive development. Through the formation of new stretching experiences, increased self - awareness and reflection my previous perspective has been overtaken by a more complex form of being aware of myself, others and the world. I refer to this process of growth as transformation. As part of my own transformational work I have conducted an inquiry into transformational growth and learning in the early academic life of university undergraduates. The result shows how accelerated adult mental development can be achieved in an academic environment ably preparing students for the workplace. This new model of education is part of a truly unique and exciting model signalling ground-breaking change for the undergraduate experience. The overhaul of a traditional BA degree in Economics into a world-class transformational programme is discussed through-out my Portfolio. Central to my broadening awareness is the challenge and nurturing required to awaken the student’s ‘internal authority’ . This involves stimulating students to take ownership for their own thinking, steering them away from the passivity and complacency of thinking through the minds of others. In doing so, the ultimate aim of renewing the BA is to narrow the developmental ‘mismatch’ which exists for m any college students between them and the world of work, by encouraging and inviting them to take on the challenge of thinking independently. Mindfulness, awareness, and personal authority are treated with reverence throughout the exploration as I consider them core parts of the students engaging with development. Engagement is construed as an active and open-minded process of awareness involving planning and reviewing one’s own goals and performance, engaging in constructive feedback, reflection and new action. I conclude with a view that the journey of adult mental development is relentless and that undergraduate education represents a crucial beginning. The value and relevance of transformational education rooted in developmental principles provides a significant opportunity in advancing development and perspectives at the start of adult life.