Sample preparation is the most critical step in proteomics as it directly affects the subset of proteins and peptides that can be reliably identified and quantified. Although a variety of efficient and reproducible sample preparation strategies have been developed, their applicability and efficacy depends much on the biological sample. Here, three approaches were evaluated for the human milk and plasma proteomes. Protein extracts were digested either in an ultrafiltration unit (filter-aided sample preparation, FASP) or in-solution (ISD). ISD samples were desalted by solid-phase extraction prior to nRPC-ESI-MS/MS. Additionally, milk and plasma samples were directly digested by FASP without prior protein precipitation. Each strategy provided inherent advantages and disadvantages for milk and plasma. FASP appeared to be the most time efficient procedure with a low miscleavage rate when used for a biological sample aliquot, but quantitation was less reproducible. A prior protein precipitation step improved the quantitation by FASP due to significantly higher peak areas for plasma and a much better reproducibility for milk. Moreover, the miscleavage rate for milk, the identification rate for plasma, and the carbamidomethylation efficiency were improved. In contrast, ISD of both milk and plasma resulted in higher miscleavage rates and is therefore less suitable for targeted proteomics.

Most hospitality institutions have increasingly moved classes online but are concerned about migrating classes and instructional content online. The concern is most Web-based models are designed to deliver the acquisition of knowledge but lack the ability to transform that knowledge into applied career skills for practical use in the industry. The purpose of this study was to test a new Web-based instructional model. The model supported delivering both the acquisition and application of knowledge. Educators, researchers, and practitioners can utilize the new model to enhance the application of career skills and enhance organizational objectives by providing just-in-time training. The new Web-based instructional model can be delivered through multiple platforms including computers, electronic devices, wireless devices and mobile devices. The application of knowledge was delivered through experiential role-play exercises delivered live to the comparison group and virtual, inside Second Life, to the treatment group. An Analysis of Co-Variance (ANCOVA) revealed a significant difference between groups with higher application scores for the students who received the role-play live compared to virtual. In addition, an analysis was conducted to explore factors to consider when examining the cost effectiveness of Web-based instructional content. Factors determined to be important were developmental costs, delivery costs, and reusability of the Web-based instruction.

Made available in DSpace on 2016-04-28T18:24:03Z (GMT). No. of bitstreams: 1 Maria Izabel Rodrigues Tognato.pdf: 1935155 bytes, checksum: d5b9e676eb6c002973db4dea833c1014 (MD5) Previous issue date: 2009-05-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This thesis aimed to contribute to the understanding of English Language teacher s work from Elementary and High School, from Public School, in the interior of Paraná State, in Brazil, through the analysis and interpretation of (re)configurations constructed on educational acting in texts produced by teachers from the state school, about this situation of work. This way, we intend to show how the educational work is (re)configured by its protagonists in texts produced by themselves about their professional activity. To develop this study, we adopt theoretical-methodical presuppositions of the sociodiscursive interactionism (ISD) (BRONCKART, 1999/2003/2007, 2004, 2006, 2008), an interdisciplinary area from Psychology, which attributes a fundamental role in the human development to the acting and the language. Considering the work as a way of acting and a factor which leads to the development and, more specifically, considering the educational work, which is institutionally regulated and that has as an intention to form the citizen, we propose to understand the connections between the language, a constitutive aspect of the human being, and the work, a central point for the understanding of the current society. This research investigates, more specifically, the connections between language and the teacher s work considering the different levels through which this work is constituted. We also assume theoretical aspects from the Sciences of the Work as the Ergonomics of the Activity (AMIGUES, 2004; SAUJAT, 2002, 2004) e, more necessarily, the Clinic of the Activity, based on the Psychology of Work (CLOT, 1999/2006; FAÏTA, 2002, 2004, 2005). This inquiry was carried through the procedure of data collection of the instruction to the double, following the perspective of the Psychology of the Work, Clinic of the Activity and Ergonomics of the Activity, taking as a basis the studies developed by Oddone et al. (1981). This procedure could allow the teacher to search for elements that supply subsidies for a critical analysis of theoretical-methodical presuppositions present at his speech, helping him to perform with a greater scientific ability on educational acting seen as work. The speeches of the investigated teachers had been analyzed according to the methodological procedures of the sociodiscursive interactionism (BRONCKART, 1999/2003/2007, 2004, 2006, 2008), considering the vision of education as a work , inspired by the Ergonomics of the Activity and the Clinic of the Activity for the interpretation of the results. We present, in this research, a synthesis of the pressupositions we assume and some questions such as: a) the work; b) educational work; c) work of English Language teacher; considering its connections in the society contemporary and with the language, beyond the c) questions about the procedure of data collection and analysis as well. Finally, we conclude the texts produced through the procedure of the instructions to the double, by the worker, can offer rich subsidies for the understanding of his situation of work. In the case of the educational work, we can not only understand what the professor carries through as what prevents his professional practice (CLOT, 1999/2006). Thus, understanding the constitutive and representative aspects of this work in a better way, it is possible to be more prepared to know how to deal with the conflicts and the choices in search for alternative solutions / Esta pesquisa teve como objetivo principal contribuir para a compreensão do trabalho do professor de Língua Inglesa de Ensino Médio e Fundamental de Escola Pública, no interior do Estado do Paraná. Para isso, desenvolvemos uma análise lingüístico-discursiva de um texto co-produzido por um professor da rede estadual de ensino em colaboração com o pesquisador e fizemos o levantamento das (re)configurações construídas sobre o agir educacional, no e por esse texto. Para chegarmos a nosso objetivo, adotamos pressupostos teórico-metodológicos do interacionismo sociodiscursivo (ISD) (principalmente em BRONCKART, 1999/2003/2007, 2004, 2006, 2008), vertente interdisciplinar da Psicologia, que atribui ao agir e à linguagem um papel fundamental no desenvolvimento humano. Além disso, assumimos aportes teóricos da Ergonomia da Atividade Francesa (AMIGUES, 2004; SAUJAT, 2002, 2004) e também da Clínica da Atividade que se fundamenta na Psicologia do Trabalho (cf. CLOT, 1999/2006; FAÏTA, 2002, 2004, 2005; ROGER, 2007; SCHELLER, 2003). A opção por esses pressupostos teórico-metodológicos de origens diversas justifica-se na medida em que todos esses autores tomam a teoria vigotskiana sobre o desenvolvimento e a concepção de linguagem de Voloshínov/Bakhtin como base de seus estudos. De um lado, o ISD nos permite tomar a linguagem como foco de nossos estudos e analisar os textos de forma bem instrumentada, pois é nos e pelos textos que se constroem as (re)configurações sobre o agir humano e, portanto, também sobre o trabalho. De outro lado, os aportes da Ergonomia da Atividade e da Clínica da Atividade, pelo maior número de pesquisas que focalizam as situações de trabalho, nos permitem construir uma visão mais ampliada sobre o trabalho docente, em suas diferentes dimensões e elementos constituintes. Além disso, é nessas disciplinas que encontramos mais suporte para a idéia de que a descrição do próprio agir, de suas dificuldades e das soluções encontradas, pelos próprios trabalhadores e, no caso, pelo professor, pode nos apontar, de modo mais claro, as diferentes formas de se (re)configurarem os elementos constitutivos desse trabalho, suas relações e suas diferentes dimensões. A nosso ver, isso permite que essas (re)configurações sejam postas em debate, aumentando o grau de possibilidades do agir do trabalhador e, assim, colaborando para seu desenvolvimento e de seu métier, considerando-se o trabalho como uma forma de agir, que tanto pode levar ao desenvolvimento quanto à amputação do poder de agir do trabalhador. A coleta de dados foi realizada por meio do procedimento das Instruções ao Sósia, em que instruções sobre como agir no trabalho são fornecidas à pesquisadora pelo professor participante da pesquisa em uma situação hipotética de substituição, co-produzindo-se o texto que foi por nós analisado no nível organizacional, enunciativo e semântico, conforme proposta de Machado (no prelo). Nele pudemos identificar diferentes figuras do agir , distribuídas em três temporalidades distintas: em momentos anteriores às aulas ministradas, no momento das aulas e em momentos posteriores a elas. Assim, o trabalho docente focalizado se mostra como um agir criativo e auto-prescritivo de preparação de aulas, de atividades e avaliações, como um agir prescritivo para o comportamento e as atividades dos alunos, como um agir organizador do meio ambiente e social; como um agir instrumentalizado, com o uso de diferentes instrumentos, dentre os quais o uso da língua portuguesa como instrumento mediador do ensino de Inglês, e, finalmente, como um agir avaliador das atividades desenvolvidas pelos alunos, envolvendo a correção de trabalhos e testes com a finalidade de retornar aos alunos um feedback sobre suas dificuldades de aprendizagem. Os resultados das análises revelam ainda que a (re)configuração do agir docente mostra seu trabalho de modo genérico, sem especificação dos procedimentos particulares de ensino em sala de aula. Os protagonistas postos em cena na atividade linguageira que se desenvolve durante o procedimento de instrução ao sósia são apenas o professor e a pesquisadora, mas muitos outros aparecem quando se trata da tematização da atividade docente: os alunos, seus pais, a supervisão, os professores da escola, outros professores em geral, o que mostra, assim, um trabalho em que a interação é um dos elementos mais fortes. Com isso, nossa pesquisa aponta para a necessidade de se desenvolver mais estudos, investigações e pesquisas sobre o trabalho do professor, pelo uso desse procedimento

Tutoring is one of the most effective instruction methods. Computer as an Intelligent Tutor is an area of research since many decades. Technology advancement in Information and Communication Technology (ICT) can be used in developing Web – based Intelligent Tutoring System (WITS), which provides individualized tutoring at the same time to large number of students geographically distributed. Intelligent Tutoring System requires knowledge representation of expert, student and instructional strategy. While web technology promises many attractive features to build web based ITS, it would still be a challenge to represent knowledge objects that are scalable, reusable and platform independent. It is required to derive generalized knowledge representation framework which can be used in developing WITS for many courses. This research work proposes an instruction System Design (ISD) model based framework in development of WITS for Control Systems. ADDIE model is selected in development of WITS. Front end analysis is conducted to identify the learning goals of a course. Proposed research work presents a Bloom - Vincenti framework for preparing learning objectives for engineering courses. Problem Based Learning (PBL) is selected as instruction strategy. Then it presents an ontology based knowledge representation framework for expert module, tutoring module, and student module. Ontology for expert module is proposed on the course structure, instruction system, instruction material ontology, and Bloom – Vincenti Taxonomy. Ontology for student module is also proposed on course structure and Bloom – Vincenti Taxonomy. Tutoring module consists of ontology about the facts of the instruction material and rule base based on the categories of engineering knowledge (Vincenti) and cognitive skill (Bloom’s Taxonomy). Proposed way of knowledge representation supports scalability, and reusability. Prototype Web – based Intelligent Tutoring System for first level course on Control Systems is developed. JAVA technology used in development of Web – based Intelligent Tutoring System (WITS), makes WITS platform independent. Web – based Intelligent Tutoring System for Control Systems is deployed at laboratory level and its efficacy is tested for first two modules of a course.

The purpose of this study was to determine teachers’ perceptions of the relationship of the Katy Management of Automated Curriculum (KMAC) system developed by Katy ISD in Katy, Texas, on planning for learning, delivery of instruction and evaluation of student learning in the classroom. KMAC is a customized, proprietary networked technology curriculum management system created for online access to curriculum and the creation and sharing of lesson plans. Data was collected from 635 teachers district-wide through an online survey. This data was used to determine whether there were differences between/among teachers and teacher leaders and between/among elementary, junior high and high school teachers in their perceived impact of the KMAC on planning for learning, delivery of instruction and evaluation of student learning. Regarding planning for learning, teachers were found to have a moderately positive perception of KMAC with teacher leaders being slightly more positive. In addition, statistically significant differences were found between grade levels with elementary teachers more positive than secondary teachers. Regarding delivery of instruction, teacher leaders again perceived a more positive relationship with KMAC than the teacher non-leaders. Statistically significant differences were also found between elementary and junior high, elementary and high school and between junior high and high school teachers, with elementary teachers being the most positive. Teachers were the least positive toward KMAC and the evaluation of student learning. While a statistically significant relationship was found in relationship to the grade level taught and evaluation, this area was admittedly weaker than the other two areas in district development and teachers’ perceptions. While the position of teacher leader seemed to impact the results in all categories, the grade level taught was found to have the greatest statistical impact on the teacher perceptions.

Increasing enrollment of students and declining availability of qualified and experienced faculty are leading to increased assessment loads of the existing faculty. Moreover, the assessment techniques are changing drastically due to the ever-increasing demand of new knowledge and abilities from the students. The tools offered by information technology can now be effectively used in enhancing the productivity of a teacher. This thesis proposes a mechanism for creating both summative and formative assessment instruments for a course in an engineering program. The assessment instruments will vary widely in nature depending on the subject. With increasing prevalence of digital devices in all walks of life a first level knowledge of digital systems is considered necessary for all engineers especially under electrical and computer engineering curricula. The first level course ‘Basics of Digital Systems’ is chosen for developing a framework of computer assisted evaluation. Creation of assessment instruments is best done in the context of an instructional system design (ISD) model. ADDIE, a generic model is chosen for the study. Bloom’s classification of levels of cognition, Vincenti’s categorization of engineering knowledge, and ‘Gronlund 2-level’ method for writing the learning objectives are integrated to create a ‘Bloom-Vincenti–Gronlund’(BVG)framework for preparing the learning objectives/assessment instruments. Developing tools for evaluation of performance of students in the assessment tests requires consideration of many issues: analysis of problems and their solution methods, errors normally committed by students, grading preferences of the instructor and feedback to students. A set of tools are developed that are able to evaluate the truth tables, state tables, excitation tables, timing diagram and VHDL codes. The developed tools are validated. The submission of the assignment and the integration of all the tools for evaluation will be more effective if they can be integrated in a learning management system (LMS). ‘MOODLE’, an open source LMS, is identified for the integration of the tools. The developed tools execute the files submitted by the students, evaluate them, and provide feedback to the students. In summary, the thesis addressed some key issues related to “assessment and evaluation of students’ performance” and proposed an integrated computer assisted system for the evaluation of students’ performance in the course ‘Basics of Digital Systems’.

A substantial amount of IT projects fail or stay challenged in meeting their targets based on project schedule, budget and system requirements.  Leading global surveys in the last decade barely indicate any improvement in the statistical performance of IT projects in spite of the fact that a lot of effort has been taken in the past to identify and fix those critical factors on which the success and failure of IT projects are generally based on. The poor trend and the underperformance of IT projects still continue. Different researches based on different approaches identify different critical factors but to a very large extent people and processes are blamed. This thesis is under taken to understand the impact of Information Systems (IS) processes and methodologies in the success and failure of IT projects. Major challenges within IS processes, such as their initial identification, implementation and organizational process awareness are identified through qualitative research methods. ‘Identification and selection of IS process and methodologies’ is identified as a major challenge within IT projects and is discussed in detail as, why and what difficulties organizations face in the process of  selecting processes and methodologies on a given IT project. A couple of methodology selection frameworks to overcome these difficulties are presented along side with their critical review and improvement. We come to a conclusion that there is a growing need for more efficient and competent methodology selection frameworks as the ones which exist today are barely complete and efficient and if they do then only to a limited and partial degree.

Antalet monoklonala antikroppar som används i läkemedel ökar kraftigt. Dessa läkemedel är dyra och risken för förfalskning är stor. Behovet att utveckla en metod för snabb och precis identifiering av monoklonala antikroppar är därför brådskande. För identifiering utfördes analyser med Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-ToF-MS), Capillary Gel Electrophoresis (CGE) and Capillary Zone Electrophoresis (CZE) på nio monoklonala antikroppar. Fokuset var att undersöka huruvida signifikanta fysiokemiska egenskaper och unika aminosyrasekvenser var närvarande och kunde urskiljas. Olika analyser med MALDI-ToF-MS användes till att både separera de monoklonala antikropparna baserat på dess fysiokemiska egenskaper, och annotera aminosyrasekvenser innehållande nyckelfragment. Med metoderna baserade på kapillärelektrofores uppnåddes också separation. CZE föredras framför CGE då mängden data som erhålls från CZE är större och provberedningen är enklare. Sammanfattningsvis utformades ett protokoll för identifieringsprocessen, vilket inleds med MALDI-ToF-MS-analyser av monoklonala antikroppar på reducerad form mot kända referenser. Därefter är en hypotes formulerad utifrån vilka antikroppar som ser mest lika ut. Slutligen analyseras dessa med CZE för fastställning av den monoklonala antikroppens identitet. / The number of monoclonal antibodies used in pharmaceuticals is increasing sharply. These medicines are expensive, and the risk of counterfeiting is high. The need to develop a method for rapid and precise identification of monoclonal antibodies is therefore urgent. For identification, analyses were performed with Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-ToF-MS), Capillary Gel Electrophoresis (CGE) and Capillary Zone Electrophoresis (CZE) on nine monoclonal antibodies. The focus was to investigate whether significant physiochemical features and unique amino acid sequences were present and could be distinguished. Various analyses with MALDI-ToF-MS were used to both separate the monoclonal antibodies based on their physicochemical properties and annotate amino acid sequences containing key fragments. With the methods based on capillary electrophoresis, separation was also achieved. CZE is preferred over CGE as the amount of data obtained from CZE is greater and sample preparation is simpler. In summary, an identification process protocol was designed and is initiated with MALDI-ToF-MS analyses of reduced-form monoclonal antibodies against known references. A hypothesis is then formulated based on which antibodies look the most similar. Finally, these are analysed by CZE to determine the identity of the monoclonal antibody.

<p></p><p>This doctoral dissertation takes the reader through a journey where applied shear rheology and flow-velocimetry are used to understand the mesoscopic factors that control the flow behavior of three microstructured fluids. Three individual protocols that measure relative physical and mechanical properties of the flow are developed. Each protocol aims to advance the particular transformation of novel soft materials into a commercial product converging in the demonstration of the real the chemical, physical and thermodynamical factors that could potentially drive their successful transformation. </p> <p> </p> <p>First, this dissertation introduces the use of rotational and oscillatory shear rheometry to quantify the solvent evaporation effect on the flow behavior of polymer solutions used to fabricate isoporous asymmetric membranes. Three different A-B-C triblock copolymer were evaluated: polyisoprene-<i>b</i>-polystyrene-<i>b</i>-poly(4-vinylpyridine) (ISV); polyisoprene-<i>b</i>-polystyrene-<i>b</i>-poly(<i>N</i>,<i>N</i>-dimethylacrylamide) (ISD); and polyisoprene-<i>b</i>-polystyrene-<i>b</i>-poly(<i>tert</i>-butyl methacrylate) (ISB). The resulting evaporation-induced microstructure showed a solution viscosity and film viscoelasticity strongly dependent on the chemical structure of the triblock copolymer molecules. </p> <p> </p> <p>Furthermore, basic shear rheometry, flow birefringence, and advanced flow-velocimetry are used to deconvolute the flow-microstructure relationships of concentrated surfactant solutions. Sodium laureth sulfate in water (SLE₁S) was used to replicate spherical, worm-like, and hexagonally packed micelles and lamellar structures. Interesting findings demonstrated that regular features of flow curves, such as power-law shear thinning behavior, resulted from a wide variety of experimental artifacts that appeared when measuring microstructured fluids with shear rheometry.</p> <p> </p> <p>Finally, the successful integration of shear rheometry to calculate essential parameters to be used in a cost-effective visualization technique (still in development) used to calculate the dissolution time of polymers is addressed. The use of oscillatory rheometry successfully quantify the viscoelastic response of polyvinyl alcohol (PVA) solutions and identify formulations changes such as additive addition. The flow behavior of PVA solutions was correlated to dissolution behavior proving that the developed protocol has a high potential as a first screening tool.</p><br><p></p>