Single-stranded DNA binding proteins (SSB) bind to single-stranded DNA (ssDNA) that is generated by molecular machines such as helicases and polymerases. SSBs play crucial roles in DNA translation, replication and repair and their importance is demonstrated by their inclusion across all domains of life. The homotetrameric E. coli SSB and the heterotrimeric human RPA demonstrate how SSBs can vary structurally, but all fulfil their roles by employing oligonucleotide/oligosaccharide binding (OB) folds. Nucleofilaments of SSB proteins bound to ssDNA sequester the ssDNA strands, and in doing so protect exposed bases, keep the ssDNA in conformations favoured by other proteins that metabolise DNA and also recruit other proteins to bind to ssDNA. This thesis focuses on the SSB from the archaeon S. solfataricus (SsoSSB), and has found SsoSSB to be a monomer that binds cooperatively to ssDNA with a binding site size of 4-5 nucleotides. Tagging ssDNA and SsoSSB with fluorescent labels allowed the real time observation of single molecule interactions during the initial nucleation event and subsequent binding of an adjacent SsoSSB monomer. This was achieved by interpreting fluorescent traces that have recorded combinations of FRET, protein induced fluorescent enhancement (PIFE) and quenching events. This novel analysis gave precise measurements of the dynamics of the first and second monomers binding to ssDNA, which allowed affinity and cooperativity constants to be quantified for this important molecular process. SsoSSB was also found to have a similar affinity for RNA, demonstrating a promiscuity not found in other SSBs and suggesting further roles for SsoSSB in the cell - possibly exploiting its capacity to protect nucleic acids from degradation. The extreme temperatures that S. solfataricus experiences and the strength of the interaction with ssDNA and RNA make exploring the application of SsoSSB for industrial uses an interesting prospect; and its rare monomeric structure provides an opportunity to investigate the action of OB folds in a more isolated environment than in higher order structures.

A simple rule for the optimal global price of carbon is presented, which captures the geo-physical, economic, and ethical drivers of climate policy as well as the effect of uncertainty about future growth of consumption. There is also a discussion of the optimal carbon budget and the amount of unburnable carbon and stranded fossil fuel reserves and a back-on-the-envelope expression are given for calculating these. It is also shown how one can derive the end of the carbon era and peak warming. This simple arithmetic for determining climate policy is meant to complement the simulations of large-scale integrated assessment model, and to give analytical understanding of the key determinants of climate policy. The simple rules perform very well in a full integrated assessment model. It is also shown how to take account of a 2 °C upper limit on global warming. Steady increases in the efficiency of labour do not affect the optimal price of carbon or the safe carbon budget, but do postpone the carbon-free era. / Series: Ecological Economic Papers

Thesis (M.Ing. (Development and Management))--North-West University, Potchefstroom Campus, 2007.

The ability to determine nucleic acid sequences is one ofthe most important platforms for the detailed study ofbiological systems. Pyrosequencing technology is a relativelynovel DNA sequencing technique with multifaceted uniquecharacteristics, adjustable to different strategies, formatsand instrumentations. The aims of this thesis were to improvethe chemistry of the Pyrosequencing technique for increasedread-length, enhance the general sequence quality and improvethe sequencing performance for challenging templates. Improvedchemistry would enable Pyrosequencing technique to be used fornumerous applications with inherent advantages in accuracy,flexibility and parallel processing. Pyrosequencing technology, at its advent, was restricted tosequencing short stretches of DNA. The major limiting factorwas presence of an isomer of dATPaS, a substitute for thenatural dATP, which inhibited enzyme activity in thePyrosequencing chemistry. By removing this non-functionalnucleotide, we were able to achieve DNA read-lengths of up toone hundred bases, which has been a substantial accomplishmentfor performance of different applications. Furthermore, the useof a new polymerase, called Sequenase, has enabled sequencingof homopolymeric T-regions, which are challenging for thetraditional Klenow polymerase. Sequenase has markedly madepossible sequencing of such templates with synchronizedextension. The improved read-length and chemistry has enabledadditional applications, which were not possible previously.DNA sequencing is the gold standard method for microbial andvial typing. We have utilized Pyrosequencing technology foraccurate typing ofhuman papillomaviruses, and bacterial andfungal identification with promising results. Furthermore, DNA sequencing technologies are not capable oftyping of a sample harboring a multitude of species/types orunspecific amplification products. We have addressed theproblem of multiple infections/variants present in a clinicalsample by a new versatile method. The multiple sequencingprimer method is suited for detection and typing of samplesharboring different clinically important types/species(multiple infections) and unspecific amplifications, whicheliminates the need for nested PCR, stringent PCR conditionsand cloning. Furthermore, the method has proved to be usefulfor samples containing subdominant types/species, and sampleswith low PCR yield, which avoids reperforming unsuccessfulPCRs. We also introduce the sequence pattern recognition whenthere is a plurality of genotypes in the sample, whichfacilitates typing of more than one target DNA in the sample.Moreover, target specific sequencing primers could be easilytailored and adapted according to the desired applications orclinical settings based on regional prevalence ofmicroorganisms and viruses. Pyrosequencing technology has also been used forclone-checking by using preprogrammed nucleotide additionorder, EST sequencing and SNP analysis, yielding accurate andreliable results. <b>Keywords:</b>apyrase, bacterial identification, dATPaS, ESTsequencing, fungal identification, human papillomavirus (HPV),microbial and viral typing, multiple sequencing primer method,Pyrosequencing technology, Sequenase, single-strandedDNA-binding protein (SSB), SNP analysis

<p>The ability to determine nucleic acid sequences is one ofthe most important platforms for the detailed study ofbiological systems. Pyrosequencing technology is a relativelynovel DNA sequencing technique with multifaceted uniquecharacteristics, adjustable to different strategies, formatsand instrumentations. The aims of this thesis were to improvethe chemistry of the Pyrosequencing technique for increasedread-length, enhance the general sequence quality and improvethe sequencing performance for challenging templates. Improvedchemistry would enable Pyrosequencing technique to be used fornumerous applications with inherent advantages in accuracy,flexibility and parallel processing.</p><p>Pyrosequencing technology, at its advent, was restricted tosequencing short stretches of DNA. The major limiting factorwas presence of an isomer of dATPaS, a substitute for thenatural dATP, which inhibited enzyme activity in thePyrosequencing chemistry. By removing this non-functionalnucleotide, we were able to achieve DNA read-lengths of up toone hundred bases, which has been a substantial accomplishmentfor performance of different applications. Furthermore, the useof a new polymerase, called Sequenase, has enabled sequencingof homopolymeric T-regions, which are challenging for thetraditional Klenow polymerase. Sequenase has markedly madepossible sequencing of such templates with synchronizedextension.</p><p>The improved read-length and chemistry has enabledadditional applications, which were not possible previously.DNA sequencing is the gold standard method for microbial andvial typing. We have utilized Pyrosequencing technology foraccurate typing ofhuman papillomaviruses, and bacterial andfungal identification with promising results.</p><p>Furthermore, DNA sequencing technologies are not capable oftyping of a sample harboring a multitude of species/types orunspecific amplification products. We have addressed theproblem of multiple infections/variants present in a clinicalsample by a new versatile method. The multiple sequencingprimer method is suited for detection and typing of samplesharboring different clinically important types/species(multiple infections) and unspecific amplifications, whicheliminates the need for nested PCR, stringent PCR conditionsand cloning. Furthermore, the method has proved to be usefulfor samples containing subdominant types/species, and sampleswith low PCR yield, which avoids reperforming unsuccessfulPCRs. We also introduce the sequence pattern recognition whenthere is a plurality of genotypes in the sample, whichfacilitates typing of more than one target DNA in the sample.Moreover, target specific sequencing primers could be easilytailored and adapted according to the desired applications orclinical settings based on regional prevalence ofmicroorganisms and viruses.</p><p>Pyrosequencing technology has also been used forclone-checking by using preprogrammed nucleotide additionorder, EST sequencing and SNP analysis, yielding accurate andreliable results.</p><p><b>Keywords:</b>apyrase, bacterial identification, dATPaS, ESTsequencing, fungal identification, human papillomavirus (HPV),microbial and viral typing, multiple sequencing primer method,Pyrosequencing technology, Sequenase, single-strandedDNA-binding protein (SSB), SNP analysis</p>

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Today, a large proportion of the world's plenteous offshore natural gas resource are stranded, flared or re-injected due to constraints pertaining to its utilisation. The major constraint in the utilisation of this resource is linked to its properties, which makes it difficult to transport or store. Although the resource presents an excellent opportunity for the Gas-to-Liquid (GTL) technology (process for converting natural gas into high energy liquid fuels with qualities that surpass the most stringent current and future clean-fuel requirements), the further processing of this resource is still impeded by high cost of transportation. However, it is believed that the emerging Floating GTL concept could offer superb opportunities to bring such offshore stranded natural gas reserves to markets by converting the gas into high quality liquid fuels, at the production sites, before it is transported using conventional oil tankers or vessels. But the question is: can this venture be profitable or economically viable? In response, an Economic Model (the EV Model) to review the economic viability of the Floating GTL option was developed. Analyses on technical and economical aspects of the floating GTL application offshore are presented with case studies on Syntroleum's and Statoil's floating GTL designs. Profitability analyses were conducted using the EV model to evaluate economic parameters such as Net Present Value (NPV), Internal Rate of Return (IRR), Discounted PayBack Period (DPBP), Profitability index (PI), Break-Even Analysis (BEA) and Scale Economies for some assumed case scenarios involving both designs. In addition, sensitivity analyses were also carried out to find the most sensitive parameters which affect the viability of the floating GTL option. The economic analyses revealed that, a modest feedstock cost (~0 - $3/MSCF), high crude oil price (that stays above $30 per barrel) and reduction trend in capital expenditure (for stand alone Floating GTL plant) up to $20,00O/BPD or lower in the next few years, will open windows for the floating GTL concept. Finally, the energy policy needed to achieve the capitalisation of the plenteous offshore stranded gas resource via floating GTL is also discussed. / Thesis (M.Ing. (Development and Management))--North-West University, Potchefstroom Campus, 2007.

Diss. (sammanfattning) Umeå : Univ., 2008. / Härtill 4 uppsatser.

Orientador: Antonio Francisco Magnoni / Resumo: O objeto desta dissertação consiste de um game didático para utilização em sala de aula como objeto formal de ensino-aprendizagem para a disciplina de Literatura do Ensino Médio da educação básica escolar brasileira, a partir da adaptação interativa da obra “Macunaíma”, do escritor modernista Mário de Andrade, dando origem ao game “Na Trilha de Macunaíma”. Para tanto, esta pesquisa tem como objetivo constituir um repertório satisfatório acerca dos aspectos teóricos, técnicos e de todos os recursos necessários para a criação e o desenvolvimento de um GDD e de um Roteiro Interativo para games didáticos. Como objetivos específicos, temos em primeiro lugar a constituição dos parâmetros pedagógicos pertinentes ao conceito apresentado para definir e parametrizar um game didático, e em segundo lugar a construção do GDD do game detalhando especialmente os aspectos pertinentes e essenciais ao Ato I (que engloba os três primeiros capítulos e outras partes interativas da narrativa original) de “Na Trilha de Macunaíma”, bem como do Roteiro Interativo também do Ato I. A intenção é apresentar uma colaboração para a problemática acerca de quais as etapas e parâmetros essenciais da criação, desenvolvimento e constituição de um GDD e de um Roteiro Interativo adequados no que se refere à criação de games didáticos a partir narrativas complexas, não lineares e multirramificadas. Como metodologia, utilizamos a pesquisa bibliográfica para os capítulos em que se desenvolve a fundamentação teórica ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This research aims to produce and present a didactic game which might satisfactorily be used in the classroom as a formal object in school learning processes for students in High School, related to the subject of Literature in Brazilian basic education, from the interactive adaptation of "Macunaíma", a novel written by the modernist Mário de Andrade, which is here presented as the game "Na Trilha de Macunaíma" (Macunaíma’s Trail). Therefore, this research aims to provide a satisfactory repertoire on the theoretical, technical and all the necessary resources for the creation and development of a GDD and an Interactive Script for didactic games. As our specific objectives, first of all this research aims to produce the pedagogical parameters pertinent to the concept presented to define and parameterize a didactic game, and then the creation and development of the Game’s GDD detailing especially the pertinent and essential aspects to Act I (that includes the three first chapters and other interactive parts of the original narrative) of the game, as well as the Interactive Script also concerning the First Act. The intention is to present a collaboration on the problematic about which are the essential stages and parameters of the creation, development and Constitution of an appropriate GDD and Interactive Script regarding to the creation of didactic games from complex, non-linear and multi-stranded narratives. As a methodology, we have used bibliographical research for the chapte... (Complete abstract click electronic access below) / Mestre

Submitted by JANAINA LEITE DE AZEVEDO null (janainalazevedo@faac.unesp.br) on 2018-01-31T00:43:27Z No. of bitstreams: 1 Dissertação_Janaina L. Azevedo_Biblioteca.pdf: 31729498 bytes, checksum: a5119b4f1d146fdcad53f7ded766d627 (MD5) / Approved for entry into archive by Maria Marlene Zaniboni null (zaniboni@bauru.unesp.br) on 2018-01-31T18:45:45Z (GMT) No. of bitstreams: 1 azevedo_jl_me_bauru.pdf: 31729498 bytes, checksum: a5119b4f1d146fdcad53f7ded766d627 (MD5) / Made available in DSpace on 2018-01-31T18:45:45Z (GMT). No. of bitstreams: 1 azevedo_jl_me_bauru.pdf: 31729498 bytes, checksum: a5119b4f1d146fdcad53f7ded766d627 (MD5) Previous issue date: 2017-03-13 / O objeto desta dissertação consiste de um game didático para utilização em sala de aula como objeto formal de ensino-aprendizagem para a disciplina de Literatura do Ensino Médio da educação básica escolar brasileira, a partir da adaptação interativa da obra “Macunaíma”, do escritor modernista Mário de Andrade, dando origem ao game “Na Trilha de Macunaíma”. Para tanto, esta pesquisa tem como objetivo constituir um repertório satisfatório acerca dos aspectos teóricos, técnicos e de todos os recursos necessários para a criação e o desenvolvimento de um GDD e de um Roteiro Interativo para games didáticos. Como objetivos específicos, temos em primeiro lugar a constituição dos parâmetros pedagógicos pertinentes ao conceito apresentado para definir e parametrizar um game didático, e em segundo lugar a construção do GDD do game detalhando especialmente os aspectos pertinentes e essenciais ao Ato I (que engloba os três primeiros capítulos e outras partes interativas da narrativa original) de “Na Trilha de Macunaíma”, bem como do Roteiro Interativo também do Ato I. A intenção é apresentar uma colaboração para a problemática acerca de quais as etapas e parâmetros essenciais da criação, desenvolvimento e constituição de um GDD e de um Roteiro Interativo adequados no que se refere à criação de games didáticos a partir narrativas complexas, não lineares e multirramificadas. Como metodologia, utilizamos a pesquisa bibliográfica para os capítulos em que se desenvolve a fundamentação teórica e em que apresentamos o estado da arte, dando preferência aos conteúdos próprios ao contexto da educação brasileira e da produção de games no Brasil. Para a criação e desenvolvimento do GDD e do Roteiro Interativo, a apresentação do produto é resultado da produção empírica calcada nos parâmetros levantados. Como resultados, este trabalho oferece tanto ao design de games quanto à educação escolar brasileira a possibilidade de promover a integração e a interdisciplinaridade entre a literatura e outras disciplinas associadas com a tecnologia, formalizando a disposição dos conteúdos pedagógicos pertinentes dentro dos parâmetros legais e pedagógicos necessários para que os games possam ser utilizados em sala de aula, bem repertórios teóricos e técnicos das áreas relacionadas ao processo de criação e desenvolvimento de games didáticos. / This research aims to produce and present a didactic game which might satisfactorily be used in the classroom as a formal object in school learning processes for students in High School, related to the subject of Literature in Brazilian basic education, from the interactive adaptation of "Macunaíma", a novel written by the modernist Mário de Andrade, which is here presented as the game "Na Trilha de Macunaíma" (Macunaíma’s Trail). Therefore, this research aims to provide a satisfactory repertoire on the theoretical, technical and all the necessary resources for the creation and development of a GDD and an Interactive Script for didactic games. As our specific objectives, first of all this research aims to produce the pedagogical parameters pertinent to the concept presented to define and parameterize a didactic game, and then the creation and development of the Game’s GDD detailing especially the pertinent and essential aspects to Act I (that includes the three first chapters and other interactive parts of the original narrative) of the game, as well as the Interactive Script also concerning the First Act. The intention is to present a collaboration on the problematic about which are the essential stages and parameters of the creation, development and Constitution of an appropriate GDD and Interactive Script regarding to the creation of didactic games from complex, non-linear and multi-stranded narratives. As a methodology, we have used bibliographical research for the chapters in which the theoretical basis is developed and in which we present the state of art, giving preference to the contents proper to the context of Brazilian education and the production of games in Brazil. For the creation and development of the GDD and the Interactive Script, the presentation of the product is a result of the empirical production based on the parameters raised. As results, this work offers both game design and Brazilian school education grounds in which the possibility of promoting integration and interdisciplinarity between literature and other disciplines associated with technology flourish, formalizing the provision of relevant pedagogical contents within the necessary legal and pedagogical parameters So that the games can be used in the classroom, as well as theoretical and technical repertoires of the areas related to the process of creation and development of didactic games.