Cloning, expression and purification of a glycosylated form of the DNA-binding protein Dps from Salmonella enterica Typhimurium

Dps, found in many eubacterial and archaebacterial species, appears to protect cells from oxidative stress and/or nutrient-limited environment. Dps has been shown to accumulate during the stationary phase, to bind to DNA non-specifically, and to form a crystalline structure that compacts and protects the chromosome. Our previous results have indicated that Dps is glycosylated at least for a certain period of the bacterial cell physiology and this glycosylation is thought to be orchestrated by some factors not yet understood, explaining our difficulties in standardizing the Dps purification process. In the present work, the open reading frame of the dps gene, together with all the upstream regulatory elements, were cloned into a PCR cloning vector. As a result, the expression of dps was also controlled by the plasmid system introduced in the bacterial cell. The gene was then over-expressed regardless of the growth phase of the culture and a glycosylated fraction was purified to homogeneity by lectin-immobilized chromatography assay. Unlike the high level expression of Dps in Salmonella cells, less than 1% of the recombinant protein was purified by affinity chromatography using jacalin column. Sequencing and mass spectrometry data confirmed the identity of the dps gene and the protein, respectively. In spite of the low level of purification of the jacalin-binding Dps, this work shall aid further investigations into the mechanism of Dps glycosylation. (C) 2008 Elsevier Inc. All rights reserved.

The surface accessibility of the glycine receptor M2-M3 loop is increased in the channel open state

Mutations in the extracellular M2-M3 loop of the glycine receptor (GlyR) alpha1 subunit have been shown previously to affect channel gating. In this study, the substituted cysteine accessibility method was used to investigate whether a structural rearrangement of the M2-M3 loop accompanies GlyR activation. All residues from R271C to V277C were covalently modified by both positively charged methanethiosulfonate ethyltrimethylammonium (MTSET) and negatively charged methanethiosulfonate ethylsulfonate (MTSES), implying that these residues form an irregular surface loop. The MTSET modification rate of all residues from R271C to K276C was faster in the glycine-bound state than in the unliganded state. MTSES modification of A272C, L274C, and V277C was also faster in the glycine-bound state. These results demonstrate that the surface accessibility of the M2-M3 loop is increased as the channel transitions from the closed to the open state, implying that either the loop itself or an overlying domain moves during channel activation.

Open Sea

Combined media on paper. 42" x 87"

The relationships between technology and open education in the development of a resilient higher education

The place of technology in the development of coherent educational responses to environmental and socio-economic disruption is here placed under scrutiny. One emerging area of interest is the role of technology in addressing more complex learning futures, and more especially in facilitating individual and social resilience, or the ability to manage and overcome disruption. However, the extent to which higher education practitioners can utilise technology to this end is framed by their approaches to the curriculum, and the socio-cultural practices within which they are located. This paper discusses how open education might enable learners to engage with uncertainty through social action within a form of higher education that is more resilient to economic, environmental and energy-related disruption. It asks whether open higher education can be (re)claimed by users and communities within specific contexts and curricula, in order to engage with an uncertain world.

An Ontology for Open Rubric Exchange on the Web

While the Internet has given educators access to a steady supply of Open Educational Resources, the educational rubrics commonly shared on the Web are generally in the form of static, non-semantic presentational documents or in the proprietary data structures of commercial content and learning management systems.With the advent of Semantic Web Standards, producers of online resources have a new framework to support the open exchange of software-readable datasets. Despite these advances, the state of the art of digital representation of rubrics as sharable documents has not progressed.This paper proposes an ontological model for digital rubrics. This model is built upon the Semantic Web Standards of the World Wide Web Consortium (W3C), principally the Resource Description Framework (RDF) and Web Ontology Language (OWL).

Contribution to Asterisk Open Source Project

With this final master thesis we are going to contribute to the Asterisk open source project. Asterisk is an open source project that started with the main objective of develop an IP telephony platform, completely based on Software (so not hardware dependent) and under an open license like GPL. This project was started on 1999 by the software engineer Mark Spencer at Digium. The main motivation of that open source project was that the telecommunications sector is lack of open solutions, and most of the available solutions are based on proprietary standards, which are close and not compatible between them. Behind the Asterisk project there is a company, Digum, which is the project leading since the project was originated in its laboratories. This company has some of its employees fully dedicated to contribute to the Asterisk project, and also provide the whole infrastructure required by the open source project. But the business of Digium isn't based on licensing of products due to the open source nature of Asterisk, but it's based on offering services around Asteriskand designing and selling some hardware components to be used with Asterisk. The Asterisk project has grown up a lot since its birth, offering in its latest versions advanced functionalities for managing calls and compatibility with some hardware that previously was exclusive of proprietary solutions. Due to that, Asterisk is becoming a serious alternative to all these proprietaries solutions because it has reached a level of maturity that makes it very stable. In addition, as it is open source, it can be fully customized to a givenrequirement, which could be impossible with the proprietaries solutions. Due to the bigness that is reaching the project, every day there are more companies which develop value added software for telephony platforms, that are seriously evaluating the option of make their software fully compatible withAsterisk platforms. All these factors make Asterisk being a consolidated project but in constant evolution, trying to offer all those functionalities offered by proprietaries solutions. This final master thesis will be divided mainly in two blocks totally complementaries. In the first block we will analyze Asterisk as an open source project and Asterisk as a telephony platform (PBX). As a result of this analysis we will generate a document, written in English because it is Asterisk project's official language, which could be used by future contributors as an starting point on joining Asterisk. On the second block we will proceed with a development contribution to the Asterisk project. We will have several options in the form that we do the contribution, such as solving bugs, developing new functionalities or start an Asterisk satellite project. The type of contribution will depend on the needs of the project on that moment.

Open Educational Resources: an opportunity for virtual learning institutions

The Universitat Oberta de Catalunya (UOC, Open University of Catalonia) is involved inseveral research projects and educational activities related to the use of Open Educational Resources (OER). Some of the discussed issues in the concept of OER are research issues which are being tackled in two EC projects (OLCOS and SELF). Besides the research part, the UOC aims at developing a virtual centre for analysing and promoting the concept of OERin Europe in the sector of Higher and Further Education. The objectives are to makeinformation and learning services available to provide university management staff,eLearning support centres, faculty and learners with practical information required to create, share and re-use such interoperable digital content, tools and licensing schemes. In the realisation of these objectives, the main activities are the following: to provide organisationaland individual e-learning end-users with orientation; to develop perspectives and useful recommendations in the form of a medium-term Roadmap 2010 for OER in Higher and Further Education in Europe; to offer practical information and support services about how to create, share and re-use open educational content by means of tutorials, guidelines, best practices, and specimen of exemplary open e-learning content; to establish a larger group ofcommitted experts throughout Europe and other continents who not only share theirexpertise but also steer networking, workshops, and clustering efforts; and to foster and support a community of practice in open e-learning content know-how and experiences.

Towards Innovative Virtual Learning in Vocational Teacher Education: Narratives as a Form of Meaningful Learning

http://www.eurodl.org/.

Open Environmental Information Upon Disclosure Request in China: The Paradox of Legal Mobilization

Taking a realist view that law is one form of politics, this dissertation studies the roles of citizens and organizations in mobilizing the law to request government agencies to disclose environmental information in China, and during this process, how the socio-legal field interacts with the political-legal sphere, and what changes have been brought about during their interactions. This work takes a socio-legal approach and applies methodologies of social science and legal analysis. It aims to understand the paradox of why and how citizens and entities have been invoking the law to access environmental information despite the fact that various obstacles exist and the effectiveness of the new mechanism of environmental information disclosure still remains low. The study is largely based on the 28 cases and eight surveys of environmental information disclosure requests collected by the author. The cases and surveys analysed in this dissertation all occurred between May 2008, when the OGI Regulations and the OEI Measures came into effect, and August 2012 when the case collection was completed. The findings of this study have shown that by invoking the rules of law made by the authorities to demand government agencies disclosing environmental information, the public, including citizens, organizations, law firms, and the media, have strategically created a repercussive pressure upon the authorities to act according to the law. While it is a top-down process that has established the mechanism of open government information in China, it is indeed the bottom-up activism of the public that makes it work. Citizens and organizations’ use of legal tactics to push government agencies to disclose environmental information have formed not only an end of accessing the information but more a means of making government agencies accountable to their legal obligations. Law has thus played a pivotal role in enabling citizen participation in the political process. Against the current situation in China that political campaigns, or politicization, from general election to collective actions, especially contentious actions, are still restrained or even repressed by the government, legal mobilization, or judicialization, that citizens and organizations use legal tactics to demand their rights and push government agencies to enforce the law, become de facto an alternative of political participation. During this process, legal actions have helped to strengthen the civil society, make government agencies act according to law, push back the political boundaries, and induce changes in the relationship between the state and the public. In the field of environmental information disclosure, citizens and organizations have formed a bottom-up social activism, though limited in scope, using the language of law, creating progressive social, legal and political changes. This study emphasizes that it is partial and incomplete to understand China’s transition only from the top-down policy-making and government administration; it is also important to observe it from the bottom-up perspective that in a realistic view law can be part of politics and legal mobilization, even when utterly apolitical, can help to achieve political aims as well. This study of legal mobilization in the field of environmental information disclosure also helps us to better understand the function of law: law is not only a tool for the authorities to regulate and control, but inevitably also a weapon for the public to demand government agencies to work towards their obligations stipulated by the laws issued by themselves.

Creating an open-books implementation framework for a maintenance-related value-based life-cycle model

This study is a qualitative action research by its nature with elements of personal design in the form of a tangible model implementation framework construction. Utilized empirical data has been gathered via two questionnaires in relation to the arranged four workshop events with twelve individual participants. Five of them represented maintenance customers, three maintenance service providers and four equipment providers respectively. Further, there are two main research objectives in proportion to the two complementary focusing areas of this thesis. Firstly, the value-based life-cycle model, which first version has already been developed prior to this thesis, requires updating in order to increase its real-life applicability as an inter-firm decision-making tool in industrial maintenance. This first research objective is fulfilled by improving appearance, intelligibility and usability of the above-mentioned model. In addition, certain new features are also added. The workshop participants from the collaborating companies were reasonably pleased with made changes, although further attention will be required in future on the model’s intelligibility in particular as main results, charts and values were all reckoned as slightly hard to understand. Moreover, upgraded model’s appearance and added new features satisfied them the most. Secondly and more importantly, the premises of the model’s possible inter-firm implementation process need to be considered. This second research objective is delivered in two consecutive steps. At first, a bipartite open-books supported implementation framework is created and its different characteristics discussed in theory. Afterwards, the prerequisites and the pitfalls of increasing inter-organizational information transparency are studied in empirical context. One of the main findings was that the organizations are not yet prepared for network-wide information disclosure as dyadic collaboration was favored instead. However, they would be willing to share information bilaterally at least. Another major result was that the present state of companies’ cost accounting systems will definitely need implementation-wise enhancing in future since accurate and sufficiently detailed maintenance data is not available. Further, it will also be crucial to create supporting and mutually agreed network infrastructure. There are hardly any collaborative models, methods or tools currently in usage. Lastly, the essential questions about mutual trust and predominant purchasing strategies are cooperation-wise important. If inter-organizational activities are expanded, a more relational approach should be favored in this regard. Mutual trust was also recognized as a significant cooperation factor, but it is hard to measure in reality.

Essays in open economy macroeconomics with borrowing frictions

Cette thèse comporte trois essais en macroéconomie en économie ouverte et commerce international. Je considère tour à tour les questions suivantes: sous quelles conditions est-il optimal pour un pays de former une union économique? (essai 1); l'augmentation de la dispersion transversale des avoirs extérieurs nets des pays est-elle compatible avec une dispersion relativement stable des taux d'investissement? (essai 2); le risque de perte de marché à l'exportation du fait de l'existence des zones de commerce préférentiel joue t-il un rôle dans la décision des pays exclus de négocier des accords commerciaux à leur tour? (essai 3). Le premier essai examine les conditions d'optimalité d'une union économique. Il s'intéresse à une motivation particulière: le partage du risque lié aux fluctuations du revenu. Dans la situation initiale, les pays ont très peu d'opportunités pour partager le risque à cause des frictions: les marchés financiers internationaux sont incomplets et il n'y pas de mécanisme pour faire respecter les contrats de crédit entre pays. Dans ce contexte, une union économique apparait comme un arrangement qui pallie à ces frictions entre les pays membres seulement. Cependant, l'union dans son ensemble continue de faire face à ces frictions lorsqu'elle échange avec le reste du monde. L'arbitrage clé dans le modèle est le suivant. D'un coté, l'intégration économique permet un meilleur partage du risque entre pays membres et la possibilité pour le partenaire pauvre d'utiliser la ligne de crédit du partenaire riche en cas de besoin. De l'autre coté, l'union peut faire face à une limite de crédit plus restrictive parce que résilier la dette extérieure est moins coûteux pour les membres l'union. De plus, le fait que le partenaire pauvre peut utiliser la limite de crédit du partenaire riche génère une externalité négative pour ce dernier qui se retrouve plus fréquemment contraint au niveau des marchés internationaux des capitaux. En conformité avec les faits observés sur l'intégration économique, le modèle prédit que les unions économiques sont relativement peu fréquentes, sont plus susceptibles d'être créées parmi des pays homogènes, et généralement riches. Le deuxième essai porte sur la dispersion des avoirs extérieurs nets et la relation avec la dispersion des taux d'investissement. Au cours des récentes décennies, la dispersion croissante des déséquilibres extérieurs et les niveaux record atteints par certaines grandes économies ont reçu une attention considérable. On pourrait attribuer ce phénomène à une réduction des barrières aux mouvements internationaux des capitaux. Mais dans ce cas, il est légitime de s'attendre à une augmentation de la dispersion au niveau des taux d'investissement; ceci, parce que le financement des besoins en investissements constitue une raison fondamentale pour laquelle les pays échangent les capitaux. Les données indiquent cependant que la dispersion des taux d'investissement est restée relativement stable au cours des récentes décennies. Pour réconcilier ces faits, je construis un modèle d'équilibre général dynamique et stochastique où les pays sont hétérogènes en raison des chocs idiosyncratiques à leurs niveaux de productivité totale des facteurs. Au niveau des marchés internationaux des capitaux, le menu des actifs disponibles est restreint à une obligation sans risque et il n'y a pas de mécanisme pour faire respecter les contrats de crédit entre pays. A tout moment, un pays peut choisir de résilier sa dette extérieure sous peine d'exclusion financière et d'un coût direct. Ce coût direct reflète les canaux autres que l'exclusion financière à travers lesquels les pays en défaut sont pénalisés. Lorsque le modèle est calibré pour reproduire l'évolution de la dispersion transversale des avoirs extérieurs nets, il produit une dispersion relativement stable des taux d'investissement. La raison principale est que les incitations que les pays ont à investir sont liées à la productivité. Avec l'intégration financière, même si les opportunités d'emprunt se sont multipliées, les incitations à investir n'ont pas beaucoup changé. Ce qui permet de générer une dispersion accrue de la position des avoirs extérieurs nets des pays avec une dispersion relativement stable des taux d'investissement. Le troisième essai analyse un aspect de l'interdépendance dans la formation des accords commerciaux préférentiels: j'examine empiriquement si le risque de diversion des exportations en faveur des pays membres des zones de commerce préférentiel est un facteur déterminant dans la décision des pays exclus de ces accords de négocier un accord à leur tour. Je construis un indicateur qui mesure le potentiel de diversion des exportations auquel font face les pays et estime un modèle probit de formation des zones de commerce préférentiel créées entre 1961 et 2005. Les résultats confirment que les pays confrontés à un plus grand potentiel de détournement des échanges sont plus susceptibles de former une zone de commerce préférentiel à leur tour.

University of Southampton Module Evaluation Form

This is the full Module Evaluation Form adopted by the University of Southampton. The latest editable file can be downloaded from the Learning and Teaching Enhancement Unit (LATEU) of the University. Included in this resource is the online version of the form for use in Blackboard, WebCT and other virtual learning environments. If you are using Blackboard, you are advised to use Internet Explorer version 6 or higher. Save the Blackboard zip archive to a local drive. Do not rename the file name. Go to the destination course area in Blackboard, open the "Control Panel" and then start the "Survey Manager" (in the "Assessment" group). Use the "Import" command to upload the zip archive. Once this is completed, rename the evaluation form which can then be added to any content area within the course using the dropdown "Add Survey" command.

Evaluation of drug physical form during granulation, tabletting and storage

An active pharmaceutical ingredient (API) was found to dissociate from the highly crystalline hydrochloride form to the amorphous free base form, with consequent alterations to tablet properties. Here, a wet granulation manufacturing process has been investigated using in situ Fourier transform (FT)-Raman spectroscopic analyses of granules and tablets prepared with different granulating fluids and under different manufacturing conditions. Dosage form stability under a range of storage stresses was also investigated. Despite the spectral similarities between the two drug forms, low levels of API dissociation could be quantified in the tablets; the technique allowed discrimination of around 4% of the API content as the amorphous free base (i.e. less than 1% of the tablet compression weight). API dissociation was shown to be promoted by extended exposure to moisture. Aqueous granulating fluids and manufacturing delays between granulation and drying stages and storage of the tablets in open conditions at 40◦C/75% relative humidity (RH) led to dissociation. In contrast, non-aqueous granulating fluids, with no delay in processing and storage of the tablets in either sealed containers or at lower temperature/humidity prevented detectable dissociation. It is concluded that appropriate manufacturing process and storage conditions for the finished product involved minimising exposure to moisture of the API. Analysis of the drug using FT-Raman spectroscopy allowed rapid optimisation of the process whilst offering quantitative molecular information concerning the dissociation of the drug salt to the amorphous free base form.