Thesis (M.S.)--West Virginia University, 2007. / Title from document title page. Document formatted into pages; contains vii, 124 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 116-124).

Submitted by Katiane Souza (katyane.souza@gmail.com) on 2016-06-05T16:57:52Z No. of bitstreams: 1 arquivototal.pdf: 2970217 bytes, checksum: d54e4990a98e77a7f5d22ad2384ffdd3 (MD5) / Made available in DSpace on 2016-06-05T16:57:52Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2970217 bytes, checksum: d54e4990a98e77a7f5d22ad2384ffdd3 (MD5) Previous issue date: 2015-12-10 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The subtribe Laeliinae comprises about 2000 species in 38 genera, exclusively neotropical and is considered the third largest family of subtribe. Cytological features most species with basic chromosome number x = 20 and karyotype evolution mainly associated with polyploidy. This study aimed performed a comparative cytogenetic analysis in 42 species of Laeliinae subtribe based on banding fluorochromes with CMA/DAPI, and estimate the nuclear DNA content of many of these species through citmetria of flow. All species presented 2n = 40 except Cattleya nobilior with 2n = 42 and the polyploid C. elongata, C. crispata, Encyclia alboxanthina, E. jenischiana, E. seidelii, Laelia gouldiana and Prosthechea faresiana (2n = 80). We observed two blocks CMA+/DAPI terminals in all species. The DNA content of species ranged from 2C = 3.45 pg in Brassavola nodosa to 2C = 7.96 pg in C. guttata. The leaf tissues of the analyzed representatives presented endoreduplication cycles in most species. Our data suggest that although it occurs an apparent macrostructural stable karyotype (2n = 40) in the species of the subtribe Laeliinae as well as the genus Cattleya studied, they present a pattern of diversification of heterochromatin consistent with the phylogenetic clusters and identify possible sinapomorphies that allow better understanding of taxonomically complex species. / A subtribo Laeliinae compreende cerca de 2000 espécies distribuídas em 38 gêneros, exclusivamente neotropical, sendo considerada a terceira maior subtribo da família. Citologicamente apresenta a maioria das espécies com número cromossômico básico x = 20 e evolução cariotípica principalmente associada a poliploidia. O presente trabalho objetivou realizada uma análise citogenética comparativa em 42 espécies da subtribo Laeliinae com base em bandeamento com fluorocromos, CMA/DAPI, além de estimar o conteúdo de DNA nuclear de várias dessas espécies através de citometria de fluxo. Todas as espécies apresentaram 2n = 40, exceto Cattleya nobilior com 2n = 42 e os poliploides C. elongata, C. crispata, Encyclia alboxanthina, E. jenischiana, E. seidelii, Laelia gouldiana e Prosthechea faresiana (2n = 80). Foram observados pelo menos dois blocos CMA+/DAPI terminais em todas as espécies, além de bandas DAPI+/CMA– proximais ou terminais em várias outras espécies. O conteúdo de DNA das espécies variou de 2C = 3,45 pg em Brassavola nodosa até 2C = 7,96 pg em C. guttata, com ciclos de endoreduplicação na maioria das espécies. Nossos dados sugerem que embora ocorra uma aparente estabilidade cariotípica macroestrutural (2n = 40) nas espécies da subtribo Laeliinae e no gênero Cattleya como um todo, em geral, o padrão de diversificação da heterocromatina foi compatível com os agrupamentos filogenéticos, identificando possíveis sinapomorfias que permitem um melhor entendimento de espécies taxonomicamente complexas.

Au cours de ma thèse, je me suis intéressée à deux modifications épigénétiques dans les cellules humaines, la méthylation de l'ADN et les modifications d'histones, et à leur relation avec deux processus cellulaires fondamentaux : l'expression génique et la réplication de l'ADN, en accordant une attention particulière aux gènes hétéro-chromatiques. J'ai étudié ces relations à travers des projets distincts en utilisant le syndrome ICF (Immunodeficiency, Centromeric instability, Facial abnormalities) comme modèle commun. Causé par un défaut épigénétique constitutif (inactivation de la DNA méthyltransfèrase 3B), le syndrome ICF (OMIM #242860) représente une source exclusive d'informations sur le rôle des modifications épigénétiques chez l'Homme. Dans un premier projet, nous avons montré que les gènes hétérochromatiques sont soumis à une hypométhylation importante et échappent à la répression transcriptionnelle dans le syndrome ICF, avec la conservation des marques d'histones. Dans une deuxième étude, nous avons observé que dans les cellules ICF la réplication du génome était plus rapide, concomitant avec une réduction de la longueur de phase S. De plus, nous avons observé un timing de réplication avancé au niveau de deux loci hétérochromatiques chez les patients ICF analysés. En parallèle de ces deux projets, j'ai mené un troisième projet, plus appliqué, centré sur le cancer colorectal.Dans ce projet, nous avons étudié la fiabilité d'un nouveau biomarqueur épigénétique (hypométhylation des loci B melanoma antigen) dans la détection des lésions pré-cancéreuses et montré que l'hypométhylation des loci BAGE est un événement précoce de la transformation des cellules du colon et est fréquent dans les adénomes histologiquement avancés. / During my PhD I studied two epigenetic modifications that occur in human cells, DNA methylation and histone modifications, and their relationship with two fundamental cellular processes: gene expression and DNA replication, with a particular attention to heterochromatic genes. I investigated this relationship in distinct projects using ICF (Immunodeficiency, Centromeric instability, Facial anomalies) syndrome as a common model. ICF syndrome (OMIM ID #242860) represents an exclusive source of information on the role of epigenetic modifications in humans, being caused by a constitutive epigenetic defect (i.e. de novo DNA methyltransferase 3B mutations). In a first project we showed that heterochromatic genes undergo hypomethylation and escape from silencing in ICF syndrome, with preservation of histone marks. In a second study we observed that whole-genome DNA replication is faster in ICF cell lines, with a concomitant shortening of the S-phase length. Besides, we observed earlier replication timing at two heterochromatic loci. In addition to these two studies, I carried out a third more applicative project focused on colorectal cancer. In this project we investigated the reliability of a new epigenetic biomarker (hypomethylation of B melanoma antigen loci) in the detection of pre-cancerous lesions and showed that BAGE loci hypomethylation is an early event in colon transformation and is frequent in histologically advanced adenomas.

Age related macular degeneration (AMD) is one of the leading causes of blindness amongst the elderly. Macular pigment (MP) in the retina has been established to protect individuals against AMD. Improving levels of MP by diet or supplements is the constant quest of clinical practitioners and researchers, thus necessitating development of instruments capable of repeatable and reliable MP measurement. Cataract, a consequence of the rising opacity levels of the lens with age is one of the other major causes of blindness in the world. Mapcatsf, a LED-based microprocessor-controlled heterochromatic flicker photometer (HFP) using photopic vision is capable of measuring the levels of MP and the opacity of the lens in terms of optical density. Test-retest measurements conducted on 83 subjects were analyzed for repeatability in macular pigment optical density (MPOD) measurements. Reliability of the lens optical density (LOD) measurements were tested and compared with those obtained at absolute scotopic thresholds for 25 individuals. A supplement study with 32 individuals both in the young (50) age groups for 6 months further established Mapcatsf’s capacity to monitor changing levels of MP in individuals. As an overall outcome, high levels of repeatability and reliability were obtained in MPOD and LOD measurements establishing Mapcatsf as an instrument for use in clinical settings in the future.

L’influence de la couleur dans les mécanismes perceptuels et attentionnels a été étudiée. Quatre couleurs (le rouge, le vert, le bleu et le jaune) ont été calibrées individuellement à travers la technique heterochromatic flicker photometry. Suivant cela, les participants ont déployé leur attention à une cible (un cercle de couleur avec une ligne orientée). Les données électrophysiologiques ont été enregistrées pendant que les sujets performaient la tâche de recherche visuelle, et les analyses ont été basées sur les potentiels évoqués (PÉs). Trois composantes des PÉs ont été examinées : la posterior contralateral positivity (Ppc); la N2pc, reflétant le déploiement de l’attention visuo-spatiale et la temporal and contralateral positivity (Ptc). Des conditions dans lesquelles la cible était bleu ou rouge, lorsque comparées à des conditions avec une cible jaune ou verte suscitaient une N2pc plus précoce. Une amplitude plus élevée est aussi observée pour les cibles rouges pour les composantes Ppc et Ptc, reflétant une sélectivité pré-attentionelle. Ces résultats suggèrent de la prudence dans l’interprétation de données comparant des cibles de différentes couleurs dans des tâches de PÉs, et ce même lorsque les couleurs sont équiluminantes. / We investigated how target color affected behavioral and electrophysiological results in a visual search task. Perceptual and attentional mechanisms were tracked using the N2pc component of the event-related potential and other lateralized components. Four colors (red, green, blue, or yellow) were calibrated for each participant for luminance through heterochromatic flicker photometry and equated to the luminance of grey distractors. Each visual display contained 10 circles, 1 colored and 9 grey, each of which contained an oriented line segment. The task required deploying attention to the colored circle, which was either in the left or right visual hemifield. Three lateralized ERP components relative to the side of the lateral colored circle were examined: a posterior contralateral positivity (Ppc) prior to N2pc, the N2pc, reflecting the deployment of visual spatial attention, and a temporal and contralateral positivity (Ptc) following N2pc. Red or blue stimuli, as compared to green or yellow, had an earlier N2pc. Both the Ppc and Ptc had higher amplitudes to red stimuli, suggesting particular selectivity for red. The results suggest that attention may be deployed to red and blue more quickly than to other colors and suggests special caution when designing ERP experiments involving stimuli in different colors, even when all colors are equiluminant.

L’influence de la couleur dans les mécanismes perceptuels et attentionnels a été étudiée. Quatre couleurs (le rouge, le vert, le bleu et le jaune) ont été calibrées individuellement à travers la technique heterochromatic flicker photometry. Suivant cela, les participants ont déployé leur attention à une cible (un cercle de couleur avec une ligne orientée). Les données électrophysiologiques ont été enregistrées pendant que les sujets performaient la tâche de recherche visuelle, et les analyses ont été basées sur les potentiels évoqués (PÉs). Trois composantes des PÉs ont été examinées : la posterior contralateral positivity (Ppc); la N2pc, reflétant le déploiement de l’attention visuo-spatiale et la temporal and contralateral positivity (Ptc). Des conditions dans lesquelles la cible était bleu ou rouge, lorsque comparées à des conditions avec une cible jaune ou verte suscitaient une N2pc plus précoce. Une amplitude plus élevée est aussi observée pour les cibles rouges pour les composantes Ppc et Ptc, reflétant une sélectivité pré-attentionelle. Ces résultats suggèrent de la prudence dans l’interprétation de données comparant des cibles de différentes couleurs dans des tâches de PÉs, et ce même lorsque les couleurs sont équiluminantes. / We investigated how target color affected behavioral and electrophysiological results in a visual search task. Perceptual and attentional mechanisms were tracked using the N2pc component of the event-related potential and other lateralized components. Four colors (red, green, blue, or yellow) were calibrated for each participant for luminance through heterochromatic flicker photometry and equated to the luminance of grey distractors. Each visual display contained 10 circles, 1 colored and 9 grey, each of which contained an oriented line segment. The task required deploying attention to the colored circle, which was either in the left or right visual hemifield. Three lateralized ERP components relative to the side of the lateral colored circle were examined: a posterior contralateral positivity (Ppc) prior to N2pc, the N2pc, reflecting the deployment of visual spatial attention, and a temporal and contralateral positivity (Ptc) following N2pc. Red or blue stimuli, as compared to green or yellow, had an earlier N2pc. Both the Ppc and Ptc had higher amplitudes to red stimuli, suggesting particular selectivity for red. The results suggest that attention may be deployed to red and blue more quickly than to other colors and suggests special caution when designing ERP experiments involving stimuli in different colors, even when all colors are equiluminant.