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Clonagem e expressão da proteína do core do vírus da hepatite C para o desenvolvimento de métodos aplicados ao diagnóstico viralSantos, Sandra Antonia Tagliavini [UNESP] 08 December 2006 (has links) (PDF)
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santos_sat_dr_araiq.pdf: 1210753 bytes, checksum: 2451e920ac425ba2bd7cc04b099878bb (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente trabalho refere-se ao desenvolvimento de duas metodologias: imobilização da proteína do core do vírus da Hepatite C (VHC) em matriz híbrida siloxano-poli (propileno óxido) como suporte sólido para ELISA e um imunossensor amperométrico para detecção de anticorpos anti-VHC. Para atingir este objetivo, o RNA-VHC extraído de amostras de soro (genótipo 1b) foi submetido à técnica de RT-PCR e posterior amplificação da seqüência de 408pb do core do VHC. Este produto foi clonado em vetor pET42a. O vetor recombinante foi introduzido em bactérias da linhagem BL21 (DES). Após o cultivo das colônias, a indução foi realizada em concentração final de 0,4mM de IPTG. As bactérias foram lisadas e as frações solúvel e insolúvel, analisadas em gel de poliacrilamida 15%, mostrando uma banda aparente de 44kDa, tamanho esperado da proteína recombinante fusionada a GST. A proteína recombinante do core foi purificada e confirmada por imunodetecção utilizando soro positivo para VHC e apresentou ausência de reatividade cruzada com amostras positivas para outras doenças infecciosas. Na primeira metodologia, a proteína do core foi imobilizada em matriz híbrida siloxano-poli (propileno óxido) preparada por sol-gel como suporte sólido em teste de ELISA para a detecção de anticorpos anti-VHC. As condições adequadas para o estabelecimento desta técnica envolveram 1,25ng da proteína/disco, conjugado com peroxidase na diluição 1:10000 e diluição do soro de 1:40. Este procedimento foi comparado ao ELISA convencional. O desenvolvimento desta matriz híbrida para imunodetecção mostrou bom desempenho, reprodutibilidade e simplicidade durante a síntese, sendo vantajoso para aplicação comercial. / The present work reports the development of two methodologies: hepatitis C vírus core protein immobilization into hybrid matrix siloxane-polypropyleneglycol prepared by sol-gel process used as solid phase in ELISA and an amperometric immunosensor for detection of antibodies anti-VHC. Toward to achieve this aim, the HCV RNA from serum (genotype 1b) was submitted to RT-PCR techniqueand subsequent amplification of the HCV core 408pb. This product was cloned into pET42a vector. The recombinant plasmid was transformed into BL21 (DES) cell line strain. Cell cultures were grown and induced with final concentration of 0,4mM of IPTG. After induction, the cell were harvest and the soluble and insoluble fractions were analyzed by polyacrilamide gel 15% showing a band with an approximate molecular weight of 44kDa, expected size for this GST-fused recombinant protein. The recombinant protein was purified and confirmed by immunological detection using HCV positive serum and showed absence of cross reactivity with positive samples for others infectious diseases. In the first methodology, the core protein immobilization into hybrid matrix siloxane-polypropyleneglycol prepared by sol-gel process was used as solid phase in ELISA for detection of antibodies anti-VHC antibody. 1,25ng protein per disc, a peroxidase conjugate diluition of 1:10000 and a serum dilution of 1:40 were adequate for the establishment of the procedure. This procedure performance of the siloxane-polypropyleneglycol discs as a matrix for immnunodetection, showed easy synthesis, good performance and reproducibility for commercial application. The second consisted on the immobilization of core protein into hybrid matrix siloxane-polypropyleneglycol prepared by sol-gel process and deposited on the pencil graphite electrode surface by dip-coating process for development of an amperometric immunosensor.
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The Relationship Between Family Recreation and Relatedness in Children and Their FamiliesBerrett, Kristen 06 December 2010 (has links)
The purpose of this study was to provide insight into the factors contributing to relatedness in children, specifically family recreation, and to look at relatedness between family members (family relatedness), and whether or not family recreation makes a contribution to this construct. Two instruments were used to collect data for this study. The Family Leisure Activity Profile (FLAP) measured family recreation involvement and satisfaction, and the Activity-Feelings States Scales (AFS) measured child and family relatedness. The sample included elementary school children between the ages of 6 years and 12 years (n1=405), and the parents of those children (n2=405). The data showed satisfaction with family recreation activities was a significant predictor of children's relatedness with their peers. The data also showed involvement in balance family recreation activities was a significant predictor of a family's relatedness with one another.
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Content Analysis of Affordances for Social and Emotional Competency Development in Third-Grade Core Reading ProgramsBrown, Lisa Trottier 01 May 2018 (has links)
The purpose of this content analysis study was to identify opportunities embedded in selected reading programs that facilitate the development of social and emotional competencies in the context of literacy instruction. These competencies have been shown to strengthen academic performance and help children learn skills that accelerate success inside and outside of the classroom. The content analysis examined three commonly used third-grade core reading programs to identify content that focuses on social and emotional competencies and instructional processes important in teaching social and emotional skills to children. The study also sought to identify content and instructional processes that might be strengthened in the curriculum to provide greater opportunities for social and emotional growth. As a part of the investigation of core reading programs, current market information was used to identify the most recent editions of the most widely used core reading programs in the U.S.
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SELF-ASSEMBLING OF NEUTRAL AND CHARGED NANOPARTICLES INTO CORE-SHELL NANOHYBRIDS THROUGH HETEROAGGREGATION WITH SIZE CONTROLUnknown Date (has links)
Core-shell nanohybrids have wide applications in pollutant degradation. In this study, core-shell nanohybrid was formed through heteroaggregation between neutral nanoparticles (i.e., hematite nanoparticles or HemNPs) and charged nanoparticles (i.e., carboxylated polystyrene nanoparticles or PSNPs). In the dispersant solution of 1 mM NaCl at pH 6.3, HemNPs were neutral and underwent favorable homoaggregation, whereas PSNPs were negatively charged and underwent no homoaggregation. When the two types of particles were mixed, homoaggregation of HemNPs and heteroaggregation between HemNPs and PSNPs took place simultaneously, forming HemNPs-PSNPs heteroaggregates. The transmission electron microscopy images of heteroaggregates show that HemNPs and PSNPs formed core-shell structure in which HemNPs were the cores and PSNPs were the shells. The size of the core-shell nanohybrids can be controlled by varying the concentration ratio of HemNPs to PSNPs. The increase of the size of charged nanoparticles resulted in larger nanohybrids. This new method has lower energy footprint than existing ones. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2019. / FAU Electronic Theses and Dissertations Collection
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Integrating the Common Core State Standards for Mathematics in a Secondary SchoolCampbell, Danielle Holmes 01 January 2017 (has links)
The problem that anchored this study was district leaders' and administrators' lack of clarity regarding teacher concerns about the Common Core State Standards for Mathematics (CCSSM) implementation in high school math. The purposes of the study were to (a) examine the perception of high school math teachers regarding the barriers for successfully implementing the CCSSM, and (b) to elicit recommendations for teacher preparation. Ely's theory of change was utilized to relate the entity of the CCSSM to this new shift in education. To better understand this phenomenon, 2 research questions accompanied this study. The research questions were geared to not only understanding teacher perceptions but also discovering strategies to assist educators with implementing the new CCSSM. Using purposeful sampling, 5 participants participated in this case study; the data collection components were an open-ended survey, interviews, and field notes. Data were analyzed by hand, using inductive reasoning and the process of coding to determine themes. The results indicated that teachers needed to know more about the standards and needed the time to gain this efficacy. Based on the themes of the study, a professional development was chosen to represent the project. Implications for positive social change are to bring awareness to teachers who are implementing the CCSSM in secondary schools, by ensuring teachers articulate consistent conversations with stakeholders, gain a form of self-efficacy, and think not only procedurally but conceptually to implement the standards. Teachers will acquire knowledge and skills to effectively educate students to become thinkers and problem solvers. This outcome will contribute to the development of college and career ready individuals.
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To what extent do core self-evaluations and coping style influence the perception of job insecurity?Låstad, Lena January 2010 (has links)
<p>Over the last few decades, increased flexibility and lack of stability in employment has made job insecurity a work stressor that keeps affecting more and more employees. This study investigates to what extent core self-evaluations influence the appraisal of job insecurity in a Swedish sample of white-collar workers (N=425). By applying the Preacher & Hayes’ macro for multiple mediation the study also tests if there is a mediating effect of coping style on the relation between core self-evaluations and job insecurity. The results show that core self-evaluations have predictive validity in relation to job insecurity. Core self-evaluations are also associated with task-based coping style. However, no mediating effect of coping style was found on the relation between core self-evaluations and job insecurity.</p> / Stockholm Stress Center / Job insecurity from a gender perspective
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Resurshantering i Dual-core klusterGustafsson, Johan, Lingbrand, Mikael January 2008 (has links)
<p>Med den nya generationen processorer där vi har flera cpu-kärnor på ett chip, så ökas prestandan genom parallell exekvering. I denna rapport presenterar vi en omvärldsstudie om allmän multiprocessorteori där vi går igenom olika tekniker för både hårdvara och mjukvara. Vi har även utfört empiriska tester på ett datorkluster, där vi har testat de två olika programmen Fluent och CFX, som utför CFD beräkningar. För varje program så har tre modeller använts för simuleringar med varierande antal beräkningsnoder. Vi har undersökt vad som är mest lönsamt, att använda en eller båda CPU-kärnorna vid de olika simuleringarna. För att testa detta har vi kört simuleringar där vi har kört med en respektive två cpu-kärnor på beräkningsnoderna. Under simuleringarna har vi samlat in mätvärden som nätverk, minne och cpu-belastning för alla noder samt exekveringstider. Dessa värden har sedan sammanställts där vi ser att ju större en modell är desto mer lönar det sig att köra med en cpu-kärna. I endast ett av våra tester har det visat sig lönsamt att använda båda cpu-kärnorna. En formel har sedan utarbetats för att påvisa skillnaderna mellan olika antal processer med en respektive två cpu-kärnor per nod. Denna formel kan appliceras för att räkna ut den totala kostnaden per simulering med hjälp av årskostnaden för de noder och licenser som används.</p>
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Region Type Checking for Core-JavaChin, Wei Ngan, Qin, Shengchao, Rinard, Martin C. 01 1900 (has links)
Region-based memory management offers several important advantages over garbage-collected heap, including real-time performance, better data locality and efficient use of limited memory. The concept of regions was first introduced for a call-by-value functional language by Tofte and Talpin, and has since been advocated for imperative and object-oriented languages. Scope memory, a lexical variant of regions, is now a core feature in a recent proposal on Real-Time Specification for Java (RTSJ). In this paper, we propose a region-based memory management system for a core subset of Java. Our region type analysis can completely prevent dangling references and thus is ready to cater for the no-dangling requirement in RTSJ. Our system also supports modular compilation, which is an important feature for Java, but was missing in recent related work. / Singapore-MIT Alliance (SMA)
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Core self-evaluations and job insecurity of employees in a government organisation / Maryka Annelize MareeMaree, Maryka Annelize January 2004 (has links)
Thesis (M.A. (Industrial Psychology))--North-West University, Vaal Triangle Campus, 2005.
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Environmental Influences On Rapid Intensity Changes In Tropical Cyclones - A Case StudyLowag, Alexander 01 January 2008 (has links)
Hurricane Bret underwent a rapid intensification (RI) and subsequent weakening between 1200 UTC August 21 and 1200 UTC August 22, 1999, before it made landfall as a category 3 hurricane on the Texas coast 12 h later. Its minimum sea-level pressure dropped 35 hPa from 979 to 944 hPa within 24 h. During this period, aircraft of the National Oceanographic and Atmospheric Administration (NOAA) flew several research missions that sampled the environment and inner core of the storm. These data sets combined with gridded data from the National Centers for Environmental Prediction's (NCEP) Global Model and the National Center for Atmospheric Research (NCAR) reanalyses are used to document the atmospheric and oceanic environments of the tropical cyclone (TC) as well as their relation to the observed structural and intensity changes. Bret's RI was linked to movement over a warm ocean eddy and high sea surface temperatures (SSTs) in the Gulf of Mexico coupled with a simultaneous decrease in vertical wind shear. SSTs at the beginning of the storm?s RI were approximately 29 degrees Celcius and steadily increased to 30 degrees Celcius as it moved northward. The vertical wind shear relaxed to less than 10 kt during this time. Mean values of oceanic heat content (OHC) beneath the storm were about 20 % higher at the beginning of the RI period than 6 h before. Cooling of near-coastal shelf waters (to between 25 and 26 degrees Celcius) by pre-storm mixing combined with an increase in vertical wind shear were responsible for the weakening of the storm. The available observations suggested that intrusion of dry air into the circulation core did not contribute to the intensity evolution. In order to quantitatively describe the influence of environmental conditions on the intensity forecast, sensitivity studies with the Statistical Hurricane Intensity Prediction Scheme (SHIPS) model were conducted. Four different cases with modified vertical wind shear and/or SSTs were studied. Differences between all cases were relatively small due to the model design, but much cooler prescribed SSTs resulted in the greatest intensity changes. Model runs with idealized environmental conditions demonstrated the model?s general lack of capability to forecast RIs and also stressed the need of more accurate SST observations in the coastal shelf regions when predicting the intensity of landfalling TCs.
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