6 resultados para Tear film stability

em Brock University, Canada


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This thesis examines the Midnight Express phenomenon focusing on the film's reception by audiences in Europe, North America, and Turkey between 1978-2003. Using and enhancing the "historical materialist approach" to film reception developed by Janet Staiger, the thesis considers the historical determinants of the film's nationally and culturally differential readings in different periods and of the transformations in those readings. The thesis argues that while Midnight Express was most likely read in the late 1970s as an attempt to reaffirm American social identity by projecting Turks as an instance of the negative Other, there has been an important shift in the reception of the film in the West during the 1990s due to the changes in the discursive contexts in which the film has been circulating. One does not observe any specific reference to Turkish prisons as a part of the issue of human rights violations in Turkey in the initial reception of the film by European and American critics, whereas these issues appear to be important constituents of a particular reception of the film in the West in the present. The thesis explains this shift by pointing to the constitution of a particular discourse on human rights violations in Turkey after 1980, and especially throughout the 1990s, which has become a part of the discursive repertoires of the Western audience. Therefore, the thesis argues that today, Midnight Express functions as a more legitimate political statement about Turkey in the eyes of some Western audiences than it had been in the 1970s. On the other hand, parallel to the increasing desire of Turkey to connect itself to the West, particularly to become a member of the European Union, one observes an immense increase in the belief in and defense against the negative effects of Midnight Express on Turkey's international representation since the 1990s. The historical and current discourses that audiences, both in Turkey and abroad, bring into play suggest that these audiences engage with Midnight Express by assuming or denying not only the subject positions constructed by the film text but also certain history-specific extra-filmic subject positions produced by other social and discursive formations.

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This thesis attempts to understand representations of death in contemporary popular film within a framework that posits mortality as a category of particular social and political importance for the way we understand both individual subjectivity and social responsibility in the postmodern cultural moment. It addresses concerns over the social organizing categories of time and space, and performs a sustained consideration of predominant themes related to the popular representation of death, such as contingency, existential.meaning, and temporal finitude. Death consciousness and social consciousness are shown to be not just intertwined, but also vitally dependent on one another, and the analyses undertaken are ultimately aimed at making these intersections explicit in order • l to think through their potential implications for challenging consumer capitalist hegemony and envisioning the possibility of progressive social change through the lens of our mortality.

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The ease of production and manipulation has made plasmid DNA a prime target for its use in gene transfer technologies such as gene therapy and DNA vaccines. The major drawback of plasmid however is its stability within mammalian cells. Plasmid DNA is usually lost by cellular mechanisms or as a result of mitosis by simple dilution. This study set out to search for mammalian genomic DNA sequences that would enhance the stability of plasmid DNA in mammalian cells.Creating a plasmid based genomic DNA library, we were able to screen the human genome by transfecting the library into Human Embryonic Kidney (HEK 293) Cells. Cells that contained plasmid DNA were selected, using G418 for 14 days. The resulting population was then screened for the presence of biologically active plasmid DNA using the process of transformation as a detector.A commercially available plasmid DNA isolation kit was modified to extract plasmid DNA from mammalian cells. The standardized protocol had a detection limit of -0.6 plasmids per cell in one million cells. This allowed for the detection of 45 plasmids that were maintained for 32 days in the HEK 293 cells. Sequencing of selected inserts revealed a significantly higher thymine content in comparison to the human genome. Sequences with high A/T content have been associated with Scaffold/Matrix Attachment Region (S/MAR) sequences in mammalian cells. Therefore, association with the nuclear matrix might be required for the stability of plasmids in mammalian cells.

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Film sequels are a pervasive part of film consumption practices and have become an important part of the decision making process for Hollywood studios and producers. This thesis indicates that sequels are not homogenous groups of films, as they are often considered, but offer a variety of story construction and utilize a variety of production methods. Three types of blockbuster sequel sets are identified and discussed in this thesis. The Traditional Blockbuster Sequel Set, as exemplified by Back to the Future (1985, 1989, 1990) films, is the most conventional type of sequel set and capitalizes on the winning formula of the first film in the franchise. The MultiMedia Sequel Set, such as The Matrix (1999,2003) trilogy, allows the user/viewer to experience and consume the story as well as the world of the film through many different media. The Lord a/ the Rings (2001, 2002, 2003) set of films is an illustration of The Saga Sequel Set where plot lines are continuous over the entire franchise thus allowing the viewer to see the entire set as a unified work. The thesis also demonstrates how the blockbuster sequel sets, such as the Pirates a/ the Caribbean (2003, 2006, 2007) franchise, restructure the production process of the Hollywood film industry.

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Endonuclease G (EndoG) is a well conserved mitochondrial nuclease with dual lethal and vital roles in the cell. It non-specifically cleaves endogenous DNA following apoptosis induction, but is also active in non-apoptotic cells for mitochondrial DNA (mtDNA) replication and may also be important for replication, repair and recombination of genomic DNA. The aim of our study was to examine whether EndoG exerts similar activities on exogenous DNA substrates such as plasmid DNA (pDNA) and viral DNA vectors, considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus (a cationic liposome transfection reagent), targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. To investigate possible effects of EndoG on viral DNA vectors, we constructed and evaluated AdsiEndoG, a first generation adenovirus (Ad5 ΔE1) vector encoding a shRNA directed against EndoG mRNA, along with appropriate Ad5 ΔE1 controls. Infection of HeLa cells with AdsiEndoG at a multiplicity of infection (MOI) of 10 p.f.u./cell resulted in an early cell proliferation defect, absent from cells infected at equivalent MOI with control Ad5 ΔE1 vectors. Replication of Ad5 ΔE1 DNA was detected for all vectors, but AdsiEndoG DNA accumulated to levels that were 50 fold higher than initially, four days after infection, compared to 14 fold for the next highest control Ad5 ΔE1 vector. Deregulation of the cell cycle by EndoG depletion, which is characterized by an accumulation of cells in the G2/M transition, is the most likely reason for the observed cell proliferation defect. The enhanced replication of AdsiEndoG is consistent with this conclusion, as Ad5 ΔE1 DNA replication is intimately related to cell cycling and prolongation or delay in G2/M greatly enhances this process. Furthermore, infection of HeLa with AdsiEndoG at MOI of 50 p.f.u./cell resulted in an almost complete disappearance of viable, adherent tumour cells from culture, whereas almost a third of the cells were still adherent after infection with control Ad5 ΔE1 vectors, relative to the non-infected control. Therefore, targeting of EndoG by RNAi is a viable strategy for improving the oncolytic properties of first generation adenovirus vectors. In addition, AdsiEndoG-mediated knockdown of EndoG reduced homologous recombination between pDNA substrates in HeLa cells. The effect was modest but, nevertheless demonstrated that the proposed role of EndoG in homologous recombination of cellular DNA also extends to exogenous DNA substrates.

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The current qualitative study examined an adapted version of the psychoeducational program, Teaching Adolescents to Think and Act Responsibly: The EQUIP Approach (DiBiase, Gibbs, Potter, & Blount, 2012). The adapted version, referred to as the EQUIP – Narrative Filmmaking Program, was implemented as a means of character education. The purpose of this study was three-fold: 1) to examine how the EQUIP – Narrative Film-making Program influenced student’s thoughts, feelings, and behaviours; 2) to explore the students’ and the teacher’s perception of their experience with the program; and 3) to assess whether or not the integrated EQUIP – Narrative Film-making Program addressed the goals of Ontario’s character education initiative. Purposive sampling was used to select one typical Grade 9 Exploring Technologies class, consisting of 15 boys from a Catholic board of education in the southern Ontario region. The EQUIP – Narrative Film-making Program required students to create moral narrative films that first portrayed a set of self-centered cognitive distortions, with follow-up portrayals of behavioural modifications. Before, during, and after intervention questionnaires were administered to the students and teacher. The student questionnaires invited responses to a set of cognitive distortion vignettes. In addition, data was collected through student and teacher interviews, and researcher observation protocol reports. Initially the data was coded according to an a priori set of themes that were further analyzed according to emotion and values coding methods. The results indicated that while each student was unique in his thoughts, feelings, and behavioural responses to the cognitive distortion vignettes after completing the EQUIP program, the overall trends showed students had a more positive attitude, with a decreased proclivity for antisocial behaviour and self-serving cognitive distortion portrayed in the vignettes. Overall, the teacher and students’ learning experiences were mainly positive and the program met the learning expectations of Ontario’s character education initiative. Based on these results of the present study, it is recommended that the EQUIP – Narrative Film-making Program be further evaluated through quantitative research and longitudinal study.