abstract: Writing scientific explanations is increasingly important, and today's students must have the ability to navigate the writing process to create a persuasive scientific explanation. One aspect of the writing process is receiving feedback before submitting a final draft. This study examined whether middle school students benefit more in the writing process from receiving peer feedback or teacher feedback on rough drafts of scientific explanations. The study also looked at whether males and females reacted differently to the treatment groups. And it examined if content knowledge and the written scientific explanations were correlated. The study looked at 38 sixth and seventh-grade students throughout a 7-week earth science unit on earth systems. The unit had six lessons. One lesson introduced the students to writing scientific explanations, and the other five were inquiry-based content lessons. They wrote four scientific explanations throughout the unit of study and received feedback on all four rough drafts. The sixth-graders received teacher feedback on each explanation and the seventh-graders received peer-feedback after learning how to give constructive feedback. The students also took a multiple-choice pretest/posttest to evaluate content knowledge. The analyses showed that there was no significant difference between the group receiving peer feedback and the group receiving teacher feedback on the final drafts of the scientific explanations. There was, however, a significant effect of practice on the scores of the scientific explanations. Students wrote significantly better with each subsequent scientific explanation. There was no significant difference between males and females based on the treatment they received. There was a significant correlation between the gain in pretest to posttest scores and the scientific explanations and a significant correlation between the posttest scores and the scientific explanations. Content knowledge and written scientific explanations are related. Students who wrote scientific explanations had significant gains in content knowledge. / Dissertation/Thesis / M.A. Curriculum and Instruction 2011

This study aims to find the answers to how an online interactive video editing tool for teachers to use would be designed. To find out the answers to this, students studying to become teachers and experienced teachers were interviewed and used for observations and usability testing of a prototype. In total there were 27 unique data gathering situations with 11 unique participants. The five teacher students who were participating were all teacher students at Linnaeus University in Växjö. The six experienced teachers have been teaching for many years and are currently lecturing teachers about new technology that can be used in the classroom. The result from interviews, observations and literature search contributed to a list of requirements which in turn became a prototype. What has been discovered is that teachers need a tool which is easy to use with interactions and functions such as adding clickable annotations to clips and creating playlists which will help teachers plan ahead and save time during lectures.

miRNAs are small regulatory RNAs, which function as post-transcriptional mRNA regulators. They direct ribonucleoprotein complexes to cognate mRNA to repress them by translational inhibition and degradation. miRNAs regulate thousands of mRNAs in mammals and have been recognized as regulatory factors in most cellular and developmental processes. Dysregulation of the miRNA pathway can lead to severe defects and diseases. Interestingly, a unique situation exists in mouse oocytes, where all the miRNA pathway components are present, yet the pathway is dispensable and nonfunctional, the molecular foundation of this phenomenon and its significance still remain unclear. In spite of the pronounced effects of the miRNA pathway in gene regulation in somatic cells, study strategies of the pathway bare limitations. Current methods for studying the activity of the miRNA pathway employ corelative studies (such as NGS) or reporter assays, which have relatively low throughput and are prone to artifacts. Here, I present design and development of a new strategy for directly monitor global miRNA pathway activity and integrity in near physiological conditions in living cells, which could also be employed in vivo for studies of mouse oocytes. The strategy is based on fluorescently tagged endogenous proteins of the...

abstract: A novel clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) tool for simultaneous gene editing and regulation was designed and tested. This study used the CRISPR-associated protein 9 (Cas9) endonuclease in complex with a 14-nucleotide (nt) guide RNA (gRNA) to repress a gene of interest using the Krüppel associated box (KRAB) domain, while also performing a separate gene modification using a 20-nt gRNA targeted to a reporter vector. DNA Ligase IV (LIGIV) was chosen as the target for gene repression, given its role in nonhomologous end joining, a common DNA repair process that competes with the more precise homology-directed repair (HDR). To test for gene editing, a 20-nt gRNA was designed to target a disrupted enhanced green fluorescent protein (EGFP) gene present in a reporter vector. After the gRNA introduced a double-stranded break, cells attempted to repair the cut site via HDR using a DNA template within the reporter vector. In the event of successful gene editing, the EGFP sequence was restored to a functional state and green fluorescence was detectable by flow cytometry. To achieve gene repression, a 14-nt gRNA was designed to target LIGIV. The gRNA included a com protein recruitment domain, which recruited a Com-KRAB fusion protein to facilitate gene repression via chromatin modification of LIGIV. Quantitative polymerase chain reaction was used to quantify repression. This study expanded upon earlier advancements, offering a novel and versatile approach to genetic modification and transcriptional regulation using CRISPR/Cas9. The overall results show that both gene editing and repression were occurring, thereby providing support for a novel CRISPR/Cas system capable of simultaneous gene modification and regulation. Such a system may enhance the genome engineering capabilities of researchers, benefit disease research, and improve the precision with which gene editing is performed. / Dissertation/Thesis / Masters Thesis Molecular and Cellular Biology 2018

Made available in DSpace on 2015-05-07T14:46:39Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1183621 bytes, checksum: b37fc0d4ae2e17bcfe0d2e1da4b88821 (MD5) Previous issue date: 2012-03-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This study aims to understand the production of Brazilian documentaries of today from one of its technical processes: the montage. We want this work to study the relationship between this stage of production and the construction of the narrative, since it is the montage process that the raw material takes shape, it is composed of images recorded for the film or image file, giving rise to the audiovisual work finalized. Undergirded by the main theories of montage and documentary filmmaking, we analyze how is the construction of their own voice in each of the films that comprise the corpus of this thesis. In the first case, we observed the relationship between the montage and construction of their own voice of the film based on the spoken word in the form of interviews and voice-over narration from the example of O fim e o princípio (2005), by Eduardo Coutinho. We take the device used in this film to apply the concepts of mounting evidence and modes of representation of Nichols (2009). In the second case we turn to the expressive use of montage features in the construction of a narrative content more poetic, this time analyzing the film Nós que aqui estamos por vós esperamos (1998), by Marcelo Masagão, where we observe the role of montage in process of "reframing" of archive images that make up the whole movie (BERNARDET, 1999). Thus, starting from two distinct forms of montage, we sought to understand the most comprehensive way possible to mount as the place of construction of the narrative in an attempt to unite the technique to the theory of media in order to give our contribution to the studies applied to contemporary documentary . / Neste estudo buscamos compreender a produção dos documentários brasileiros dos dias atuais a partir de um dos seus processos técnicos: a montagem. Queremos neste trabalho estudar a relação existente entre esta etapa da produção e a construção da narrativa, uma vez que é na montagem que o material bruto ganha forma, seja ele composto por imagens gravadas para o filme ou por imagens de arquivo, dando origem à obra audiovisual finalizada. Embasados pelas principais teorias da montagem e do cinema documentário, analisamos como se dá a construção da voz própria em cada um dos filmes que compõem o corpus desta dissertação. No primeiro caso, observamos a relação entre a montagem e a construção da voz própria do filme com base na palavra dita sob a forma de entrevistas e narração em voz over, a partir do exemplo de O fim e o princípio (2005), de Eduardo Coutinho. Aproveitamos o dispositivo adotado neste filme para aplicarmos os conceitos de montagem de evidência e modos de representação de Nichols (2009). No segundo caso nos atemos ao uso expressivo dos recursos de montagem na construção de uma narrativa de teor mais poético, desta vez analisando o filme Nós que aqui estamos por vós esperamos (1998), de Marcelo Masagão, onde observamos o papel da montagem no processo de ressignificação das imagens de arquivo que compõem todo o filme (BERNARDET, 1999). Assim, partindo de duas formas distintas de montagem, buscamos compreender da maneira mais abrangente possível a montagem enquanto lugar de construção da narrativa, em uma tentativa de unir a técnica à teoria do audiovisual no intuito de dar nossa contribuição para os estudos aplicados ao documentário contemporâneo.

Edição semidiplomática de corpus selecionado a partir do códice ?Ofícios do Vice-Rei para o Governador da Capitania 1793/98?, visando a fornecer fonte fidedigna para futuros estudos filológicos, lingüísticos, históricos e culturais do período. Trata-se também de um estudo mais detalhado desse códice catarinense que contextualiza historicamente os documentos que o compõem, assim como aborda aspectos paleográficos, codicológicos e diplomáticos com o propósito de responder a questionamentos e hipóteses surgidos durante a edição acerca de sua composição, criação e origem. Este trabalho descreve codicológica e paleograficamente o códice em geral, apresenta uma demonstração de diferentes punhos e procura levantar traços de natureza fonológica de um manuscrito dentro do códice. Consta também deste trabalho uma tentativa de identificação tipológica mais adequada através da composição de um verbete descritivo do corpus. / Semidiplomatic edition of the selected corpus from the codex ?Oficios do Vice Rei para o Governador da Capitania 1793/98? (Memorandums from the Vice-king to the Governor of the Province 1793/98?) seeking to provide a trustworthy source to further philological, linguistical, historical and cultural studies of such period. It is also a thorough study of this Catarinense (from Santa Catarina) codex that put its documents into a historical perspective. It touches its paleographic, codicological and diplomatic aspects aiming to address questionings and hypotheses raised during the editing process concerning its composition, how it was assembled, and its origin. This work describes the codex general paleography and codicology. It presents a demonstration of different handwritings and attempts to raise traces of phonological aspects from the codex. This project also proposes a more adequate typological identification of the documents, through descriptive summaries of each memo.

A produção de proteínas terapêuticas recombinantes compreende moléculas complexas e de alto valor agregado, incluindo a enzima glucocerebrosidase (GCase). Sua deficiência resulta na Doença de Gaucher, passível de tratamento por meio da terapia de reposição enzimática. A forma ativa da GCase recombinante usada na terapia apresenta resíduos terminais de manose expostos no seu perfil de glicosilação. Perfil este que espera-se ser reproduzido por meio da construção de uma linhagem de Pichia pastoris com um padrão de glicosilação humanizado, por meio da deleção de dois genes envolvidos no sistema de glicosilação da levedura: alg3 e och1, responsáveis pela posterior hiper-manosilação característica desse organismo. Assim, a expressão da GCase será usada como modelo no desenvolvimento desta linhagem de Pichia pastoris que permita a expressão de glicoproteínas com um perfil humanizado específico de glicosilação. Além da produção da linhagem mutante pela técnica de CRISPR/Cas9, propomos a construção de duas linhagens controle: uma expressando a proteína GCase para análise do seu padrão selvagem de glicosilação em P. pastoris e outra expressando a proteína Cas9 de Streptoccocus pyogenes (SpCas9). A linhagem P. pastoris/GCase foi construída testando-se duas sequências sinal de secreção diferentes: fosfatase alcalina (PHO1) e albumina humana (Alb). Resultados de western blot mostraram a GCase no lisado celular e baixos níveis de proteína secretada no sobrenadante de cultura, sendo mais expresso na linhagem contendo a sequência PHO1. A linhagem P. pastoris/SpCas9 foi construída e a enzima SpCas9 foi detectada via western blot no lisado celular após indução com metanol. Para a produção da linhagem com padrão de glicosilação humanizado propôs-se a deleção dos genes alg3 e och1 e a inserção, pela via de reparo por recombinação homóloga (HDR), de marcas de resistência aos antibióticos higromicina ou canamicina. Para tal, propusemos a construção de dois vetores finais de expressão do sistema CRISPR/Cas9 em P. pastoris, cada um contendo a enzima SpCas9 e os RNAs guia (gRNAs) para deleção do gene alg3 ou och1, e também a construção de dois fragmentos para HDR contendo o gene de resistência ao antibiótico flanqueado por regiões de 1Kb de homologia com a região de deleção do gene alg3 ou och1. A construção dos vetores e fragmentos para HDR foram inicialmente feitas por meio de técnicas de clonagem clássica. No entanto, apesar de inúmeras tentativas, resultados de PCR e sequenciamento mostraram o insucesso das construções. Partiu-se então para a técnica de Gibson Assembly®, através da qual os dois fragmentos para HDR foram construídos. Porém, os vetores de expressão contendo SpCas9 e os gRNAs ainda apresentam dificuldades na sua construção. Esforços ainda estão sendo feitos para a construção dos vetores e consequente tentativa de estabelecimento das linhagens mutantes. O sucesso no estabelecimento de um sistema de expressão de proteínas heterólogas com este padrão de glicosilação humano específico permitirá a obtenção e possível comercialização da GCase em sua forma terapêutica. Além disso, permitirá possíveis edições genômicas futuras para um padrão de maior complexidade de glicosilação humanizado, criando uma plataforma nacional para produção de outras glicoproteínas terapêuticas de interesse biotecnológico. / The production of therapeutic recombinant protein comprises complex and high valued molecules, including the glucocerebrosidase enzyme (GCase). Its deficiency results in Gaucher Disease, susceptible of treatment by enzymatic replacement therapy. The active form of recombinant GCase employed in therapy presents exposed terminal mannose residues in its glycosylation pattern. We hope to reproduce such pattern by constructing a Pichia pastoris strain with a specific human glycosylation pattern through the deletion of two genes involved in yeast glycosylation system, alg3 and och1, responsible for the final hyper-mannosylation characteristic of this organism. Therefore, the expression of GCase will be a case model for the development of the recombinant Pichia pastoris strain that could allow the expression of glycoproteins with a specific humanized glycosylation profile. Despite the establishment of the mutant strain using the CRISPR/Cas9 technique, we propose the construction of two control strains: one expressing the GCase protein for analysis of its wild type glycosylation pattern and another one expressing the Cas9 protein from Streptoccocus pyogenes (SpCas9). The P. pastoris/GCase strain was constructed testing two different secretion signal sequences: alkaline fosfatase (PHO1) and human albumin (Alb). Western blot results have shown GCase in cell lysate and in low expression levels in culture supernatant, being more expressed in the strain containing the PHO1 signal sequence. P. pastoris/SpCas9 strain was constructed and SpCas9 enzyme was detected via western blot in cell lysate after the induction with methanol. To produce the strain with the humanized glycosylation pattern, the deletion of alg3 and och1 genes was proposed along with the insertion, by homology directed repair pathway (HDR), of hygromycin and kanamycin antibiotics resistance marks. In order to do so, we have proposed the construction of two final expression vectors of the CRISPR/Cas9 system in P. pastoris, each one containing SpCas9 enzyme and the guide RNAs (gRNAs) for deletion of alg3 or och1, and also the construction of two fragments for HDR containing the antibiotics resistance gene flanked by 1Kb regions of homology with the deleted regions of alg3 or och1. Vectors and HDR fragments constructions were initially performed using classic cloning techniques. However, despite numerous tries, PCR and sequencing results have shown the failure of the constructions. Then, we moved on to the Gibson Assembly® technique, through which the two HDR fragments were built. Still, the expression vectors containing SpCas9 and the gRNAs presented difficulties in its assembly. Efforts continue to be made to successfully construct the remaining vectors and to establish the mutant lineage. Success in the establishment of a heterologous protein expression system with specific human glycosylation pattern will allow the obtainment and possible commercialization of the therapeutic form of GCase. Furthermore, it will also allow possible future genomic editing to a high complexity human glycosylation pattern, creating a national platform for the production of other therapeutic glycoproteins of biotechnological interest.

São muitas as parcerias no cinema, mas a que relaciona o diretor de cena ao diretor de fotografia é certamente a principal. A proposta desta dissertação de mestrado é analisar o estilo de fotografia criado por Eduard Tissé em três dos filmes que trabalhou com Eisenstein. A metodologia teórica da pesquisa baseia-se em Roland Barthes a partir da análise do artefato primeiro do cinema, que é o fotograma. As dimensões sociais ligadas ao materialismo dialético e suas aplicações ao filme, na forma como o espectador promove a leitura de sentidos, preocupavam o diretor soviético. Com isso evidencia-se o princípio conceitual de Eisenstein, desenvolvido pela célula de montagem, na visão da sua missão sócio-cultural em busca da fotografia do conflito. / There are many partnerships in the movie business, but that between director and director of photography is certainly the main one. The purpose of this dissertation is to analyze the style of photography created by Eduard Tissé in three of the movies he worked with Eisenstein. The theoretical research methodology is based on Roland Barthes from the analysis of the first film artifact: the frame. The social dimensions related to the dialectical materialism and its applications to the cinema - the way the audience reads the film sense - worried the soviet director. With that, the conceptual principle developed by Eisenstein is highlighted in search of the conflict cinematography.

A montagem cinematográfica foi um advento que mudou os rumos da história do audiovisual. O processo de edição e montagem possui uma importância ímpar na constituição da linguagem audiovisual. Modificada desde Méliès e aperfeiçoada ao longo do tempo, a montagem cinematográfica se tornou um dos pilares sobre os quais se sustentam os estudos de diversos autores dentro da história do Audiovisual, especificamente na história e teoria do cinema. Através da leitura de inúmeras dessas obras, em conjunto com a análise fílmica, podemos constatar que a manipulação do tempo e do espaço durante o processo de montagem de uma obra audiovisual permite a construção de múltiplos sentidos discursivos. Considerando essa relevância da montagem na construção da narrativa fílmica, esse projeto visa refletir sua importância discursiva por meio da análise de um recurso empregado constantemente na montagem: o faux raccord. A evidência do aparato e a adição de sentidos na etapa da pós-produção através da utilização do faux raccord faz com que esse elemento seja fulcral na análise da importância da montagem discursiva. Assim, essa pesquisa busca postular uma precisa definição para o termo, bem como situá-lo na produção teórica e audiovisual ao longo da história. Compreendendo seus usos e aplicações e averiguando de que modo esse elemento gesta nos filmes sentidos discursivos e propicia ao espectador a percepção dessa construção de sentido através da evidência do corte. / Film editing was a invention that changed the course of audiovisual\'s history. The process of editing has a unique importance in the creation of audiovisual language. Modified since Méliès and improved over time, film editing became one of the pillars in film studies, specifically in history and film theory. By reading many theoretical works, combined with film analysis, we note that the manipulation of time and space during the film editing process of an audiovisual work allows the construction of multiple discursive meanings. Considering the importance of editing in the construction of film narrative, this project is a reflection of discursive importance through the analysis of a resource constantly used in film editing: faux raccord. Therefore, this research seeks to postulate a precise definition for the term, and place it in the theoretical and audiovisual production throughout history. Understanding its uses and applications, and reflecting how this element generates discursive meanings in films and provides the spectator the perception of meaning construction through the evidence of the cut.

En 1967, sous l’influence du jeune cinéma européen, Hollywood accède à son tour à la modernité : le « Vieil Hollywood » laisse place à un « Nouvel Hollywood ». Ce mouvement de renouvellement du cinéma américain, symboliquement inauguré par Bonnie and Clyde d’Arthur Penn et Le lauréat de Mike Nichols, est, explique-t-on, le fait d’une génération nouvelle de cinéastes, parmi lesquels, outre Penn et Nichols, Francis Ford Coppola, Brian De Palma, George Lucas, William Friedkin,Michael Cimino, Martin Scorsese, Robert Altman, Bob Rafelson, Hal Ashby... Il est aussi le fait d’une génération nouvelle de monteurs, qui, en complicité avec leurs réalisateurs, éprouvèrent de nouvelles formes d’expressivité esthétique et narrative, sans craindre de contrevenir aux règles conventionnelles instituées par leurs aînés ; parfois même en réaction contre ces règles. On peut notamment citer, parmi ces fidèles et précieux collaborateurs, Dede Allen, Walter Murch, Sam O’Steen, Verna Fields, Alan Heim, Paul Hirsch, Robert C. Jones, Richard Marks, ou encore Ralph Rosenblum. Nous veillons à déterminer, dans cette thèse, la part de responsabilité, tant collective qu’individuelle, de cette nouvelle génération de monteurs dans le mouvement de renouvellement esthétique du cinéma américain. Et, au-delà des personnes, d’apprécier ce style de montage néohollywoodien, d’évaluer son importance dans la constitution du style neuf des films américains des années 1970, de même que l’étendue de ses innovations. / In 1967, under the influence of European art films, Hollywood underwent important changes in the course of the 1960s : « Old Hollywood » was taken over by a « New Hollywood ». This process of renewal of American cinema, which has been symbolically launched by Arthur Penn’s Bonnie and Clyde and Mike Nichols’ The Graduate, is due, it has been said, to the arrival of a new – mostly film-school educated – generation of filmmakers, such as Francis Ford Coppola, Brian De Palma,George Lucas, William Friedkin, Michael Cimino, Martin Scorsese, Robert Altman, Bob Rafelson or Hal Ashby. It is also due to the arrival of a new generation of film editors who did not fear to challenge the doctrine of classical Hollywood editing and break the established rules in order to take a chance and try new ways of telling a story. These films editors are, amongst others, Dede Allen, Walter Murch, Sam O’Steen, Verna Fields, Alan Heim, Paul Hirsch, Robert C. Jones, Richard Marks, or Ralph Rosenblum. Our thesis examines the share of responsability, on both a collective and individual level, of this new generation of film editors in this process of renewal of American cinema in the 1960s and 1970’s. Its goal is, more generally, to determine and appreciate this new editing style, realize its importance and its influence on the Hollywood Renaissance style, as well as the wide range of its innovations.