Com o propósito de discorrer em que medida a telenovela brasileira se aproxima do docudrama ao desenvolver narrativas híbridas entre a ficção e a realidade, este trabalho investiga a origem e os mecanismos de funcionamento desse gênero ainda pouco explorado pela comunidade científica. Partindo de uma periodização, já proposta e consolidada em outras investigações, avançamos na hipótese de que a telenovela brasileira, na contemporaneidade, acelerou o processo de hibridação do formato, iniciado desde sua origem, ao combinar e mesclar linguagens e formatos de diversas ordens. O docudrama é um gênero audiovisual híbrido composto por convenções formais do documentário com o drama, mais especificamente o melodrama. A partir da proposta de leitura de documentários, formulada pelo cineasta francês Roger Odin, que nos permite identificar, no nível da imagem, uma estrutura estilística típica dos documentários, pensamos numa proposta de leitura específica de docudramas. Através da análise de programas como Linha Direta e Por Toda Minha Vida, identificamos características comuns ao gênero: uso intensivo de referentes (fotografias, imagens de arquivo, depoimentos) misturados com a dramatização de fatos reais. Tal qual um fait divers, uma espécie de crônica adaptada ao romance-folhetim e que fez grande sucesso entre os leitores no século XIX, os docudramas apresentam temáticas que priorizam tragédias, crimes, acidentes e assuntos de repercussão popular. Com base nessas razões, a pesquisa segue para a telenovela brasileira para verificar em que medida esse formato de ficção seriada aproxima-se do docudrama ao trazer fatos e pessoas \"reais\" para a ficção. Esta investigação parte da premissa de que a telenovela brasileira encontra-se num período que denominamos de \"novelas híbridas\" que apresenta narrativas em sintonia com o período histórico, político, social e econômico pautados por um processo de globalização em curso nas quais as fronteiras comerciais e culturais tornam-se mais tênues. Nesse contexto, a telenovela brasileira, reconhecida nacional e internacionalmente, pela qualidade técnica e de criação, viaja pelo mundo representando a cultura do país e divulgando seu modo de produção dessa modalidade de ficção seriada que, cada vez mais, cria histórias baseadas em fatos reais na contramão da ficção: a antificção. / With the purpose to discuss the extent to which Brazilian telenovela approaches is close to docudrama while developing fiction and reality hybrid narratives, this paper investigates the origin and the mechanisms of this genre still largely unexplored by the scientific community. Starting with a timeline, already proposed and consolidated in other investigations, we further hypothesized that the Brazilian telenovela, in contemporary times, accelerated the hybridization of the format, launched from its origin to combine and merge languages and formats of various orders. The docudrama is an audiovisual hybrid genre composed of formal conventions of the documentary with the drama, specifically the melodrama. From the reading of the proposed documentary, made by French filmmaker Roger Odin, which allows us to identify the level of the image, a stylistic structure typical of documentaries, we proposed a specific reading of docudramas. Through the analysis of shows such as Linha Direta and Por Toda Minha Vida, characteristics common to the genre: the intensive use of referents (photographs, archival images, testimonies) mixed with the dramatization of actual events were identified. Like a fait divers, a kind of chronicle adapted to paper-novel and that was a big hit among readers in the nineteenth century, the docudramas have themes that prioritize tragedies, crimes, accidents and issues of popular repercussion. Based on these reasons, the search goes to the Brazilian telenovela to see to what extent the format of serial fiction approaches the docudrama to bring facts and real people into fiction. This research assumes that the Brazilian telenovela is set in a period we call \"hybrid telenovelas\" that present narratives in line with the historical, political, social and economic period guided by an ongoing globalization process in which commercial and cultural borders become more tenuous. In this context the Brazilian telenovela, recognized nationally and internationally for the quality and technical creation, travels the world representing the country\'s culture and spreading its mode of production of this type of fiction serial that increasingly creates stories based on real facts counter to the fiction: the anti-fiction.

Background: Over the relatively short period of its evolutionary history, Boechera (Brassicaceae) has undergone rapid radiation that has produced 70+ morphologically distinct, sexual diploids. However, reproductive isolation has moved more slowly than morphological divergence in this group and the diploids appear to hybridize frequently where they coexist. Boechera duchesnensis appears to be the result of hybridization between its putative parents Boechera thompsonii and Boechera formosa. Objectives: The objectives of this study are to (i) analyze and document genetic diversity patterns in the population structure, - including allelic and heterozygosity frequencies - of B. thompsonii and B. formosa in concert with their geographic distribution to determine clustering relationships within these populations, (ii) confirm and expand the morphological characteristics of B. thompsonii and B. formosa, as initially proposed in the literature, including pollen and trichome structure using Scanning Electron Microscopy (SEM) to confirm ploidy level and to determine whether both putative parent species share morphological characteristics with their apomictic diploid offspring, and (iii) use genetic and morphologic evidence to show that B. thompsoii and B. formosa are, in fact, the parents of B. duchesnensis by comparing the genetic diversity patterns, population structure, and morphological characteristics of B. duchesnensis, to those of its proposed putative parents (B. thompsonii and B. formosa) and to confirm that B. duchesnensis shares characteristics of Boechera. Methods: Microsatellite data from 14 loci previously identified in Boechera were used to reexamine the current classifications and taxonomic foundations of three Boechera spp. GenAlEx 6.501 (Peakall and Smouse, 2006, 2012) was used to analyze genetic population structures of two divergent sexual diploids in the genus Boechera: B. thompsonii and B. formosa and to later compare those with the population structure of B. duchesnensis. Geographicaldata were plotted using ArcGIS 10.1 (Esri, 2012) to map heterozygosity distribution. Cluster analysis was run with STRUCTURE 2.3.3 (Pritchard et al., 2000; Falush et al., 2003, 2007) and distribution of allelic diversity and heterozygosity was subsequently compared within each taxon and correlated with geographic distribution characteristics. Resultant data were then compared with B. duchesnensis data to document genetic diversity patterns, population structure, and morphological characteristics. Key Results: Analysis of genetic diversity patterns, allelic distribution of the populations, and heterozygosity of B. thompsonii and B. formosa across their geographic range identified four genetically distinct clusters within B. thompsonii, and one genetically distinct cluster in B. formosa. Allelic frequencies in all four discrete population clusters of B. thompsonii and in one discrete population cluster of B. formosa were close to values found in species on the decline. Reproductive isolation, genetic variability, and allelic frequencies were determined, specimen elevations reported, and morphological characteristics reported in the literature were confirmed and expanded. A codominant genetic analysis performed for 14 different loci for B. duchesnensis against those of its parents showed that B. duchesnensis inherits alleles from both putative parents and confirms B. thompsonii and B. formosa as the parents of B. duchesnensis. Observed levels of heterozygosity of B. thompsonii and B. formosa were lower than expected levels and lower than those of other outcrossing diploids. The mean overall observed heterozygosities for each cluster were determined and documented by geographic location. A substantially higher level of observed heterozygosity in B. duchesnensis (Ho = 0.908) consistent with genetic fixation of a heterozygote and apomixis, supports hybridization as a speciation mechanism and apomixis as a mode of reproduction accounting for genotypic and phenotypic diversity. Morphological characteristics, especially those of pollen and trichomes were confirmed, expanded, and documented with SEM imagery. Discussion: This study provides an analysis of the genetic diversity patterns inherent in the population structure, allelic frequencies, allelic variation among individuals of the rare sexual diploids B. thompsonii, B. formosa, and the apomictic diploid B. duchesnensis in correlation with their geographic distribution. There is an implication of a reproductive barrier, within populations of the same species, that contributes to genetic isolation between clusters. I analyze the tendency of reduced heterozygosity to lead to genetic fixation, reproductive isolation, and how the heightened heterozygosity supports the classification of B. duchesnensis as an apomict. Assessing potential populations that might exist based on similar characteristics could possibly provide inferences about where future research might find similar examples of this hybridization. Reproductive isolation is hypothesized to limit gene flow between identified clusters of B. thompsonii and B. formosa exacerbating low observed heterozygosity levels and low allelic frequency levels. Population studies and cluster analysis have implications for offering future conservation strategies for both taxa.

Endometriosis is one of the most common gynecological diseases as it is thought to affect up to 15% of the female population. Characterized by the growth and proliferation of endometrial tissue outside of the uterine cavity, it is a complex condition with varying degrees of severity and can affect multiple regions of the body with symptoms ranging from a total lack of symptoms to debilitating pain and infertility. The most accepted theory of how endometriosis initiates is that of retrograde menstruation; however, approximately 90% of women with unobstructed fallopian tubes are thought to have some menstrual debris in the peritoneal cavity. Therefore, this theory does not explain in full why endometriosis occurs in some but not all women who experience retrograde bleeding. Genetic factors are thought to play a major role in the pathogenesis of endometriosis as women with a family history are 5 to 10 times more likely to develop the disease. The goal of this study was to determine if common chromosomal aberrations in the form of additions, deletions, or regions of loss of heterozygosity that may contribute to the establishment or progression of the disease are present in a population of endometriosis patients. DNA was isolated from the peripheral blood of endometriosis patients and endometriosis tissue biopsies, and it was analyzed using oligonucleotide based array comparative genomic hybridization. The results suggest that an addition on chromosome 17p13.3 may play a role in the biological mechanisms involved in endometriosis as it was identified in 75% of the DNA samples obtained from the peripheral blood and 100% of the DNA samples obtained from the tissue biopsies. This chromosomal imbalance is of particular interest as it is located in a region that harbors the tumor suppressor gene, hypermethylated in cancer-1 (HIC-1), whose aberrant expression has been reported in multiple cancers. Endometriosis has long been thought of as a benign disease despite its malignant characteristics, and individuals with endometriosis have been demonstrated to have an increased chance of developing ovarian cancer. This was the first study to examine the DNA from endometriosis patients using oligonucleotide based array comparative genomic hybridization to investigate genetic abnormalities in endometriosis. The findings may provide a novel target for future therapeutic options as well as indicate a link between endometriosis and cancer that has not been previously reported.

The objective of this study was to identify genes that are differentially regulated during diapause when compared with nondiapausing pupae in Sarcophaga crassipalpis. The results of a Suppression Subtractive Hybridization procedure was used to indentify genes that are differentially regulated in both diapause and nondiapausing states while suppressing genes that are common to both states. Randomly picked colonies from both subtractive libraries were isolated and the inserts sequenced. The sequences were analyzed using the bioinformatics tools NCBI, BlastX, Clustal W, etc. Out of 384 clones, 59 genes were found to be upregulated during diapause and 37 genes were found to be upregulated during a nondiapause pupal stage, no genes were found to be expressed commonly in both the diapause and nondiapause constructed libraries.

The oral pathogen Streptococcus mutans is a known etiological agent for dental root decay and coronal caries. It has been hypothesized, but not yet proven, that S. mutans expression of virulence genes in dental plaque may be influenced by its interaction with co-aggregating partners, notably Fusobacterium nucleatum and Actinomyces viscosus. Investigation of the suitability of mixed cultures of S. mutans with F. nucleatum versus S. mutans with A. viscosus proved that A. viscosus was a better target in the present laboratory setting. Furthermore, A. viscosus, a causative agent of mandible osteomyelitis and endocarditis, has been shown to have direct interaction ability with S. mutans. DNA microarray analysis was used in the present study to investigate the influence of co-aggregation with A. viscosus on the expression of S. mutans genes. Microarrays have been used successfully in the analysis of differential gene expression in S. mutans as a function of culture conditions, such as in biofilms versus planktonic states. This technology however, has not yet been applied to the analysis of homologous versus heterologous cultures. The present study was conducted in order to identify potential problems associated with the application of microarray analysis to mixed cultures. The data obtained encourage the further testing of microarrays for the analysis of heterologous cultures of oral bacteria.

Biological invasions are a global threat to aquatic biodiversity. Of particular concern are invasive freshwater fishes because they have high establishment rates, and introductions can result in the displacement and extirpation of native species through a range of processes including competition and hybridization. Though it is well known that invasive freshwater fishes commonly spread following introduction events, little is known about how fast and far they may move. Additionally, observations of hybridization involving invasive stream fishes have been linked to elevated turbidity; however, the extent to which impaired water clarity influences reproductive isolation among invasive and native species remains poorly understood. To better understand how invasive freshwater fishes disperse, and how turbidity affects reproductive isolation between native and non-native species, I carried out a series of three related studies. First, I evaluated genetic variation across the native and invasive ranges of red shiner (Cyprinella lutrensis), throughout the United States. Second, I characterized genetic variation and clinal stability across a hybrid swarm involving native blacktail shiner (Cyprinella venusta stigmatura) and invasive red shiner in the Upper Coosa River Basin (UCRB), USA. Third, I examined whether turbidity influences pre-mating social interactions between invasive red shiner and native blacktail shiner. MtDNA haplotypes from native range populations of red shiner form four divergent lineages and suggest that introduced populations in the western and eastern US originate from dissimilar genetic lineages. I also recovered a previously undescribed lineage of Cyprinella that has been cryptically introduced into the western US. Examination of the hybrid swarm in the UCRB revealed that the proportion of hybrids increased between 2005 and 2011, and that the hybrid swarm is continuing to expand both upstream and downstream. Under turbid conditions, I found that pre-mating social interactions increased, and that native blacktail shiner females are especially likely to interact with invasive red shiner males. Localized control or removal may be effective in managing non-native red shiner; further monitoring, however, is needed to help identify additional factors contributing to hybrid swarm movement. Furthermore, integrating knowledge of species behavior into management planning could help deter the further establishment and spread of invasive red shiner. / acase@tulane.edu

Doctor of Philosophy / Benign tumours grow by expanding and displacing the surrounding tissues, while malignant tumours replace and destroy the surrounding tissues by invasion. Although there is extensive literature on mechanisms of tumour invasion and metastasis, with an emphasis on angiogenesis, adhesion, degradation of the extracellular matrix and migration, an important question not clearly addressed by the literature, but nonetheless approached in this thesis, is that of the fate of normal cells during tissue replacement by migrating invasive malignant cells. Earlier work in the laboratory where this PhD candidature was carried out, investigated the effect of osteosarcoma cells on endothelium. In contrast to the expected angiogenic effect of malignant cells for endothelium, it was found that the human osteosarcoma cell line (SAOS-2) induced apoptosis in human umbilical vein endothelial cells (HUVEC) in contact dependent manner (McEwen et al., 2003). It was suggested that apoptosis of endothelium by malignant tumour cells may facilitate tumour invasion and metastasis (McEwen et al., 2003), and one of the aims of the current study was to extend these findings to include human gingival fibroblasts (HGF) and human umbilical artery smooth muscle cells (HUASMC). The major finding of this thesis was that SAOS-2 induced a reduction in the apparent cell culture density of HGF and HUASMC in a contact-dependent manner. The SW480 colorectal carcinoma cell line did not have any clear effect upon the apparent stromal cell culture density of either HGF or HUASMC, suggesting that the effect under investigation was tumour cell line specific. Surprisingly and in contrast to the similar effect reported for endothelium (Chen et al., 2005; McEwen et al., 2003), the effect of SAOS-2 upon HGF and HUASMC was not due to stromal cell apoptosis. Apoptosis was ruled out as a possible mechanism for the reduced apparent culture density under study, by using widely accepted methods which are dependent upon intermucleosomal fragmentation of DNA, the permeability of plasma membranes to dyes in advanced apoptosis and necrosis, phosphatidylserine translocation as well as inhibitor studies blocking both caspase dependent and independent pathways. While apoptosis was not demonstrated, the possibility emerged that reduced apparent stromal cell culture density reflected fusion events rather than the simple removal of cells as had been earlier reported for HUVEC (McEwen et al., 2003). This idea was supported by reduced SAOS-2 circularity in co-culture. Confocal microscopy of cells pre-labelled with fluorescent dyes further supported this idea, with dual-labelling as evidence of cell fusion. Although occasional homotypic fusion of stromal cells was seen, heterotypic fusion of stromal cells with SAOS-2 was much more prevalent. Time lapse microscopy was performed to further characteristic cell fusion in co-cultures, and revealed multiple transient fusions between SAOS-2 and HGF. To work towards determining the biological relevance of the key observation, two stable SAOS-2 GFP clones were generated for future planned studies using human gingival explants and nude mice. Importantly, the clones were similar to native SAOS-2 with regard to alkaline phosphatise expression and reducing apparent stromal cell culture density. Transient fusions between HGF and SAOS-2, may be a mechanism for cooption of stromal cells into the malignant process, facilitating tumour invasion. Additionally, heterocellular fusion of SAOS-2 with stromal cells may facilitate immune evasion, while it seems likely that despite the absence of an identical activity in SW480 cells, other malignant tumour cells may also express similar activity.

<p>A series of novel methods for detection of known sequence variants in DNA, in particular single nucleotide polymorphism, using padlock probes and rolling circle replication are presented. DNA probes that can be circularized – padlock probes – are ideal for rolling circle replication. Circularized, but not unreacted probes, can generate powerful signal amplification by allowing the reacted probes to template a rolling circle replication (RCR) reaction. However, when hybridized and ligated to a target DNA molecule with no nearby ends, the probes are bound to the target sequence, inhibiting the RCR reaction is. This problem can be solved by generating a branched DNA probe with two 3’ arms such that the probes may be circularized while leaving the second 3’ arm as a primer for the RCR reaction. We describe how T4 DNA ligase can be used for efficient construction of DNA molecules having one 5’ end but two distinct 3’ ends that extend from the 2’ and 3’ carbons of an internal nucleotide. An even stronger approach to circumvent the topological problem that can inhibit RCR is to restriction digest the template downstream of the padlock recognition site. By using Phi 29 DNA polymerase with efficient 3’ exonuclease and strand displacement activity, the template strand can then be used to prime the RCR reaction. The amplified molecule is contiguous with the target DNA, generating an anchored localized signal. The kinetics of the reaction was investigated by following the reaction in real-time using molecular beacon probes. Localized RCR signal were obtained on DNA arrays, allowing detection of as little as 104-105 spotted molecules, of either single- or double-stranded M13 DNA, in a model experiment. We have also established a serial rolling circle amplification procedure. By converting rolling circle products to a second and even third generation of padlock probes the signal was amplified thousand-fold per generation. This procedure provides sufficient sensitivity for detection of single-copy gene sequences in 50 ng of human genomic DNA, and large numbers of probes were amplified in parallel with excellent quantitative resolution.</p>

<p>The great difference in the level of chloroplast variation and its geographic structure among the three main species studied here demonstrates that forest species do not form a homogeneous group. Hazel shows a genetic structure similar to many other thermophilous species and this structure, in combination with fossil evidence, indicates that the post-glacial colonisation of most of Europe originated in a refugium in western France while the Balkan and Italy were colonised from a south-eastern refugium.</p><p>In sallow and silver birch the chloroplast DNA variation and its structure does not fit with a scenario of glacial restriction to southern refugia and survival at intermediate latitudes is suggested for both species. The chloroplast DNA variation in silver birch suggests the presence of one western and one eastern European post-glacial colonisation route and limited contribution of southern populations in the colonisation of the rest of Europe. Unique haplotypes by the Ural Mountains indicates the possibility of a separate glacial origin of these populations.</p><p>The study of chloroplast DNA in species closely related to sallow and silver birch indicate that extensive hybridisation and cytoplasmic gene flow occurs within both the Salix and Betula genera in Europe. The nuclear and chloroplast phylogenies of 14 Betula species were not in complete agreement with each other or with the classical division of the Betula genus into subgenera or sections. The phylogenetic structure implies that hybridisation has played a role in the evolution of the Betula genus.</p><p>This thesis focuses on the chloroplast DNA variation in three forest tree genera: Corylus, Betula and Salix. Chloroplast PCR-RFLP is used to evaluate the post-glacial history of hazel, Corylus avellana, silver birch, Betula pendula and sallow, Salix caprea and to explore the possibility of introgression in the Salix and Betula genera. In addition, the chloroplast matK gene, its flanking regions and the nuclear ADH gene were used to study the phylogenetic relationships within the Betula genus.</p>

<p>Speciation is the process in which reproductive barriers evolve between populations. In this thesis I examine how sexual signals contribute to the maintenance, reinforcement or breakdown of reproductive barriers.</p><p>Male pied flycatchers (<i>Ficedula hypoleuca</i>) and collared flycatchers (<i>F. albicollis</i>) differ in song and plumage traits. However, where the two species coexist, several pied flycatchers sing a song resembling the collared flycatcher (mixed song). Mixed song is not caused by introgression from the collared flycatcher but is due to heterospecific copying. Mixed song provokes aggressive behaviour in collared flycatcher males and leads to heterospecific pairing and maladaptive hybridization. </p><p>The species differences in song were found to be larger in an old than a young hybrid zone. This was due to a reduction in the frequency of mixed song in the pied flycatcher and a divergence in the song of the collared flycatcher. Apparently, mixed song causes maladaptive hybridization, which over time leads to reinforcement of reproductive barriers by a song divergence.</p><p>Previous studies have shown that a character displacement in male plumage traits reinforces species barriers. Hence both plumage and song divergence reduce the incidence of hybridization. The evolution of male plumage traits has been so rapid, or selection has been so strong that rapidly evolving molecular markers are unable to trace it.</p><p>Hybrid females mate with a male of the same species as their father. Previous studies have shown that females use male plumage traits controlled by genes linked to the sex chromosomes (the Z) in species recognition. An association between preference and a sex-linked trait through the paternal line may render reinforcement of reproductive barriers more likely.</p><p>In conclusion, sexual signals are affected by species interactions that cause breakdown or reinforcement of reproductive barriers.</p>