Plasmid DNA (pDNA) vectors are the current conventional technology driving therapeutic gene transfer, whether for use toward mal/nonfunctional gene replacement, DNA vaccination, or production of therapeutic proteins in mammalian cells. However, the conventional pDNA vector suffers from several safety and efficiency limitations: 1) it imparts adverse immune responses to bacterial sequences required for maintenance and amplification in prokaryotes; 2) its bioavailability can be compromised due to size; and 3) it may be genotoxic due to its potential to integrate into the host chromosome and yield an oncogenic event. In this study we have constructed a robust in vivo bacterial platform for the production of bacterial sequence-free linear covalently closed (LCC) DNA vectors, termed DNA Ministrings, through the manipulation and application of bacteriophage-encoded recombination systems. Phage N15 and PY54 lysogenize their bacterial hosts as a linear plasmid with covalently closed ends (LCC plasmid). LCC morphology is conferred by the phage-encoded telomerase via a single cleaving-joining reaction of the perfect palindrome target site. This system was exploited to generate DNA Ministring vectors, encoding only the gene(s) of interest and necessary complementary eukaryotic expression/enhancement genetic elements that are devoid of unwanted bacterial sequences and are linearized through a single in vivo enzymatic reaction. The tel and telN prokaryotic telomerase (protelomerase) genes were amplified from PY54 and N15 lysates, respectively, and cloned into a bacterial vector that expresses the gene under control of the temperature sensitive bacteriophage ?? CI857 repressor that confers conditional expression from ?? pL/pR promoters. This regulatory circuit was integrated into a RecA+ lacZ+ E. coli K-12 strain via homologous recombination, where successful recombinants were disrupted for the lacZ gene. Recombinant cells are capable of conditional expression of the phage-derived telomerase enzymes by shifting the temperature to >37 ??C. Phage P1-derived Cre recombinase was applied as a positive control, since its functionality in generating DNA minicircle vectors has been previously shown. A multi-purpose 342 bp target site termed Super Sequence (SS) that possesses the Cre, Flp, Tel, and TelN target sites in addition to two flanking SV40 enhancer sequences was cloned into two different sites of a GFP expression eukaryotic pDNA vector. The amplification of this DNA vector through telN / tel or cre expressing Recombinant E. coli cells (R-cells) generated bacterial sequence-depleted (LCC) DNA Ministring and (CCC) Minicircle vectors, respectively, as evidenced by digestion patterns of the purified vector. Transfection efficiency of these vectors was assessed in rapidly dividing human ovarian cancer and in relatively slowly dividing human embryonic kidney cell lines. In vitro experiments with DNA Ministrings in human cells lines resulted in significantly higher transfection efficiency, bioavailability, and cytoplasmic diffusion levels compared to the parental plasmid precursor and isogenic DNA Minicircle counterparts. The safety of the LCC DNA vector conformation, with respect to insertional genotoxicity, was assessed by forcing LCC pDNA vectors into bacterial and human genomic DNA. The integration of LCC DNA into bacterial and human host genomic DNA resulted in chromosomal DNA disruptions at site of integration, loss of genome stability, and subsequent cell death. LCC integration-induced apoptotic cell death and natural elimination of the integrant from human cell population improves the safety profile of DNA Ministrings by eliminating integrants following the potential genotoxic side effects of undesired vector integration into the host genome.

Esta dissertação realiza uma análise do livro A Máquina do Tempo, escrito por H.G. Wells em 1895, e sua transposição fílmica homônima realizada por George Pal em 1960 dentro do escopo do gênero ficção-científica. São analisadas as mudanças que foram realizadas no processo de transposição da literatura para o cinema. Este trabalho discute inicialmente as características do gênero ficção-científica na literatura, como o gênero se diferencia de outros gêneros como a literatura fantástica, por exemplo, e quais são os elementos que compõem a identidade do gênero ficção-científica. Em um segundo momento investigamos os problemas que o gênero ficção-científica apresenta quando transposto a mídia cinematográfica. Entre os fatores investigados estão como a verossimilhança funciona na literatura de ficção-científica e como o leitor se relaciona com ela. Também se analisou como o gênero utiliza o conhecimento científico para criar seus universos ficcionais, e as mudanças que os realizadores fílmicos às vezes precisam fazer para tornar o universo literário ficcional aceitável ao espectador em termos de verossimilhança, considerando que o que é verossímil na literatura não é necessariamente verossímil no cinema, especialmente na literatura de ficção-científica, um gênero que possui uma ligação íntima com a ciência e o progresso. Para finalizar essa pesquisa analisa a relação entre o gênero ficção-científica e o cinema no que diz respeito aos problemas que o gênero apresenta ao cinema em termos de tecnologia. Em outras palavras, o gênero literário ficção-científica traz aos realizadores fílmicos problemas e desafios que exigem a pesquisa e o desenvolvimento de novas tecnologias e técnicas visando resolvê-los. / The present work analyzes the book The Time Machine, written by H.G.Wells in 1895, as well as its transposition into a homonymous movie made by George Pal in 1960 as part of the science fiction genre. This work initially discusses the characteristics of the science fiction genre in literature, the way the genre differs from others such as the fantastic literature, for instance, and the fundamental elements of the science fiction genre. It then investigates the problems the science fiction genre presents when transposed to film media. Among the investigated factors are the problem of verisimilitude in science fiction literature and how the reader responds to it. This includes discussing the way the genre uses scientific knowledge to create its fictional universe, and the changes, in terms of verisimilitude, moviemakers need to consider when the fictional literary text is transferred into film. What sometimes is plausible in literature not always seems plausible in movies, especially in regard to science fiction literature, a genre that has a intimacy with science and progress. To conclude, this research analyzes the relation between the science fiction genre and film concerning the challenges the genre presents when transposed into another medium in terms of technology. These challenges require research and the development of new procedures for the cinematographic medium.

Esta dissertação realiza uma análise do livro A Máquina do Tempo, escrito por H.G. Wells em 1895, e sua transposição fílmica homônima realizada por George Pal em 1960 dentro do escopo do gênero ficção-científica. São analisadas as mudanças que foram realizadas no processo de transposição da literatura para o cinema. Este trabalho discute inicialmente as características do gênero ficção-científica na literatura, como o gênero se diferencia de outros gêneros como a literatura fantástica, por exemplo, e quais são os elementos que compõem a identidade do gênero ficção-científica. Em um segundo momento investigamos os problemas que o gênero ficção-científica apresenta quando transposto a mídia cinematográfica. Entre os fatores investigados estão como a verossimilhança funciona na literatura de ficção-científica e como o leitor se relaciona com ela. Também se analisou como o gênero utiliza o conhecimento científico para criar seus universos ficcionais, e as mudanças que os realizadores fílmicos às vezes precisam fazer para tornar o universo literário ficcional aceitável ao espectador em termos de verossimilhança, considerando que o que é verossímil na literatura não é necessariamente verossímil no cinema, especialmente na literatura de ficção-científica, um gênero que possui uma ligação íntima com a ciência e o progresso. Para finalizar essa pesquisa analisa a relação entre o gênero ficção-científica e o cinema no que diz respeito aos problemas que o gênero apresenta ao cinema em termos de tecnologia. Em outras palavras, o gênero literário ficção-científica traz aos realizadores fílmicos problemas e desafios que exigem a pesquisa e o desenvolvimento de novas tecnologias e técnicas visando resolvê-los. / The present work analyzes the book The Time Machine, written by H.G.Wells in 1895, as well as its transposition into a homonymous movie made by George Pal in 1960 as part of the science fiction genre. This work initially discusses the characteristics of the science fiction genre in literature, the way the genre differs from others such as the fantastic literature, for instance, and the fundamental elements of the science fiction genre. It then investigates the problems the science fiction genre presents when transposed to film media. Among the investigated factors are the problem of verisimilitude in science fiction literature and how the reader responds to it. This includes discussing the way the genre uses scientific knowledge to create its fictional universe, and the changes, in terms of verisimilitude, moviemakers need to consider when the fictional literary text is transferred into film. What sometimes is plausible in literature not always seems plausible in movies, especially in regard to science fiction literature, a genre that has a intimacy with science and progress. To conclude, this research analyzes the relation between the science fiction genre and film concerning the challenges the genre presents when transposed into another medium in terms of technology. These challenges require research and the development of new procedures for the cinematographic medium.

Salicylic acid (SA) is an important plant hormone with a wide range of effects on plant growth and metabolism. Plants lacking SA exhibit enhanced susceptibility to pathogens. SA plays important signaling roles in resistance against biotrophic and hemi- biotrophic phytopathogens. It is synthesized in plastids along two pathways, one involving phenylalanine ammonia lyase (PAL) and the other isochorismate synthase (ICS). In Arabidopsis , during immune response most SA is synthesized through the ICS-dependent pathway, but clearly an ICS-independent pathway also exists. Several SA effector proteins have been identified and characterized which mediate downstream SA signaling. This includes SABP, a catalase, SABP2, a methyl salicylate esterase, SABP3, a carbonic anhydrase, NPR1 (nonexpressor of pathogenesis-related 1), NPR3 (a NPR1 paralog), and NPR4 (another NPR1 paralog). NPR3 and NPR4 regulate the turnover of NPR1, a process which plays a key role in activating defense gene expression. The role of SA in abiotic stress signaling is gradually becoming clearer. Various components of SA signaling in biotic stress also appear to impact abiotic stress signaling.

Au cours des dernières décennies, le Québec a connu une augmentation significative du nombre d’élèves ayant des défis particuliers dans les classes ordinaires du primaire. Cette diversification des profils d’apprentissage a complexifié l’enseignement, qui vise la réussite de tous les élèves indépendamment de leurs besoins spécifiques. Les pratiques pédagogiques et les ressources matérielles jouent un rôle crucial pour favoriser l’accès, la participation et la progression pour tous les élèves. Toutefois, les enseignants ne semblent pas suffisamment préparés, tant dans leur formation initiale que continue, pour mettre en place des pratiques dites accessibles dans la classe. Cette étude vise à mieux comprendre comment la formation courte s’appuyant sur le processus de planification universelle (PPU) peut soutenir les enseignants du primaire afin de planifier et mettre en œuvre la conception universelle de l’apprentissage (CUA) et les aides technologiques (AT) dans la classe. Plus spécifiquement, cette étude vise à concevoir un dispositif de formation à proposer aux enseignants, à documenter la réalisation des étapes du PPU par les enseignants et à décrire les apports et les limites du dispositif de formation quant au soutien apporté par ce dernier aux enseignants du primaire au Québec. À partir de la littérature recensée, un dispositif de formation a été élaboré et proposé à cinq enseignants du primaire. Les enseignants ont suivi une formation en ligne d’environ une heure de façon individuelle avant de participer à une rencontre de travail collectif. Les résultats de cette recherche, s’appuyant sur une méthodologie qualitative, montrent que le dispositif de formation courte offre un soutien prometteur aux enseignants. Celui-ci a contribué à changer les perceptions des enseignants en ce qui concerne les difficultés à apprendre orientant le regard vers l’identification des obstacles plutôt que des déficiences. De plus, le PPU a été utile pour soutenir la planification et la mise en œuvre de la CUA et des AT par les enseignants. Enfin, la collaboration, l’accompagnement et la conception des ressources de formation selon les principes de la CUA ont joué un rôle pour soutenir la compréhension et le développement d’habiletés au cours de cette formation. / Over the last few decades, Quebec has seen a significant increase in the number of students with special challenges in regular primary school classes. This diversification of learning profiles has complicated teaching, which aims at the success of all students regardless of their specific needs. Educational practices and material resources play a crucial role in promoting access, participation and progress for all students. However, teachers do not appear to be sufficiently prepared, both in their initial and continuing training, to implement so-called accessible practices in the classroom. This study aims to better understand how short training based on the Planning for all learners (PAL) can support primary school teachers in planning and implementing the Universal design of learning (UDL) and assistive technology (AT) in the classroom. More specifically, this study aims to design a training to be proposed to teachers, to document the achievement of the PAL stages by teachers and to describe the contributions and limits of the training system in terms of the support provided by the latter to primary school teachers in Quebec. Based on the literature collected, a training scheme was developed and proposed to five primary school teachers. Teachers took about an hour of online training individually before participating in a collective work meeting. The results of this research, based on a qualitative methodology, show that the short-term training system offers promising support to teachers. This has helped change teachers’ perceptions of learning difficulties by looking to identify obstacles rather than disabilities. Most of all, the PPU has been useful in supporting the planning and implementation of UDL and ATs by teachers. Finally, the collaboration, support and design of training resources in accordance with the UDL principles played a role in supporting understanding and skills development during this training.