Development of high specific strength components, with internal cellular structure, for several applications

Dissertação de mestrado em Engenharia Mecânica

Analysis of a boundary protein delimiting chromatin domains in yeast

SUMMARY The expression state of a eukaryotic gene depends in part on its location in the chromosome. This position effect results from the organization of eukaryotic genomes into discrete functional domains, defined by local differences in chromatin structure. The expression of genes within each domain appears to be defined and maintained by the concerted action of regulatory elements such as promoters, enhancers, silencers and locus control regions. Individual domains may be bordered by boundary elements that separate regions of permissive and silent chromatin. When located next to chromosomal elements such as telomeres, genes can be subjected to epigenetic silencing. In yeast, this is mediated by the propagation of the SIR proteins from telomeres towards more centromeric regions. Particular transcription factors can protect downstream genes from silencing when tethered between the gene and the telomere, and they may thus act as chromatin domain boundaries. Here we have studied one of these transcription factors, CTF-1, that binds directly histone H3. A deletion mutagenesis localized the barrier activity to CTF-1 histone-binding domain. A saturating point mutagenesis of this domain identified several amino-acid substitutions that similarly inhibited the boundary and histone-binding activities. Chromatin immunoprecipitation experiments indicated that the barrier protein efficiently prevents the spreading of SIR proteins, and that it separates domains of hypoacetylated and hyperacetylated histones. Together, these results suggest a mechanism by which proteins such as CTF-1 may interact directly with histone H3 to prevent the propagation of a silent chromatin structure, thereby defining boundaries of permissive and silent chromatin domains. RESUME L'expression des gènes eucaryotes dépend en partie de leur localisation sur les chromosomes. Cet effet de position résulte de l'organisation des génomes eucaryotes en domaines fonctionnels, définis par des changements locaux au niveau de la structure de la chromatine. Dans chacun de ces domaines, l'expression des gènes est définie et maintenue par l'action concertée de différents éléments régulateurs tels que les promoteurs, les amplificateurs, les silenceurs et les locus control régions. Ces domaines peuvent être entourés par des éléments barrière, séparant les régions de chromatine répressive des régions permissive pour l'expression des gènes. Lorsqu'ils se situent à proximité d'éléments chromosomiques comme les telomères, les gènes peuvent être réprimés de manière épigénétique. Chez la levure, cette répression est établie par la propagation des protéines SIR depuis les télomères vers les régions centromériques. Certains facteurs de transcription peuvent empêcher la répression d'un gène, lorsqu'ils sont placés entre ce gène et le télomère. Nous avons étudié un de ces facteurs, CTF-1, qui a la particularité de lier directement l'histone H3. La délétion de certaines parties de CTF-1 a permis de déterminer que la région responsable de l'activité barrière correspond au domaine d'interaction avec H3. Plusieurs mutations points effectuées dans ce domaine inhibent à la fois l'activité barrière et la capacité de lier H3. Des expériences d'immuno-précipitation de la chromatine indiquent que la protéine barrière CTF-1 prévient efficacement la propagation des protéines SIR et sépare des domaines contenant des histones hypo-acétylées de ceux constitués d'histones hyper-acétylées. Ces résultats suggèrent que CTF-1 interagit directement avec l'histone H3 pour empêcher la propagation de la chromatine répressive, délimitant ainsi des domaines de chromatine permissive et des domaines de chromatine silencieuse.

Structure et évolution thermique de la croûte supérieure des chaînes de subduction : approches thermologique et modélisation numérique, dans les North Cascades (U.S.A) et de la zone de faille Motagua (Guatemala)

Cette thèse cible l'étude de la structure thermique de la croûte supérieure (<10km) dans les arcs magmatiques continentaux, et son influence sur l'enregistrement thermochronologique de leur exhumation et de leur évolution topographique. Nous portons notre regard sur deux chaînes de montagne appartenant aux Cordillères Américaines : Les Cascades Nord (USA) et la zone de faille Motagua (Guatemala). L'approche utilisée est axée sur la thermochronologie (U-Th-Sm)/He sur apatite et zircon, couplée avec la modélisation numérique de la structure thermique de la croûte. Nous mettons en évidence la variabilité à la fois spatiale et temporelle du gradient géothermique, et attirons l'attention du lecteur sur l'importance de prendre en compte la multitude des processus géologiques perturbant la structure thermique dans les chaînes de type cordillère, c'est à dire formées lors de la subduction océanique sous un continent.Une nouvelle approche est ainsi développée pour étudier et contraindre la perturbation thermique autour des chambres magmatiques. Deux profiles âge-elevation (U-Th-Sm)/He sur apatite et zircon, ont été collectées 7 km au sud du batholithe de Chilliwack, Cascades Nord. Les résultats montrent une variabilité spatiale et temporelle du gradient géothermique lors de l'emplacement magmatique qui peut être contrainte et séparé de l'exhumation. Durant l'emplacement de l'intrusion, la perturbation thermique y atteint un état d'équilibre (-80-100 °C/km) qui est fonction du flux de magma et de ia distance à la source du magma, puis rejoint 40 °C/km à la fin du processus d'emplacement magmatique.Quelques nouvelles données (U-Th)/He, replacées dans une compilation des données existantes dans les Cascades Nord, indiquent une vitesse d'exhumation constante (-100 m/Ma) dans le temps et l'espace entre 35 Ma et 2 Ma, associée à un soulèvement uniforme de la chaîne contrôlé par l'emplacement de magma dans la croûte durant toute l'activité de l'arc. Par contre, après ~2 Ma, le versant humide de la chaîne est affecté par une accélération des taux d'exhumation, jusqu'à 3 km de croûte y sont érodés. Les glaciations ont un triple effet sur l'érosion de cette chaîne: (1) augmentation des vitesses d'érosion, d'exhumation et de soulèvement la où les précipitations sont suffisantes, (2) limitation de l'altitude contrôlé par la position de Γ Ε LA, (3) élargissement du versant humide et contraction du versant aride de la chaîne.Les modifications des réseaux de drainage sont des processus de surface souvent sous-estimés au profil d'événements climatiques ou tectoniques. Nous proposons une nouvelle approche couplant une analyse géomorphologique, des données thermochronologiques de basse température ((U-Th-Sm)/He sur apatite et zircon), et l'utilisation de modélisation numérique thermo-cinématique pour les mettre en évidence et les dater; nous testons cette approche sur la gorge de la Skagit river dans les North Cascades.De nouvelles données (U-Th)/He sur zircons, complétant les données existantes, montrent que le déplacement horizontal le long de la faille transformante continentale Motagua, la limite des plaques Caraïbe/Amérique du Nord, a juxtaposé un bloc froid, le bloc Maya (s.s.), contre un bloque chaud, le bloc Chortis (s.s.) originellement en position d'arc. En plus de donner des gammes d'âges thermochronologiques très différents des deux côtés de la faille, le déplacement horizontal rapide (~2 cm/a) a produit un fort échange thermique latéral, résultant en un réchauffement du côté froid et un refroidissement du côté chaud de la zone de faille de Motagua.Enfin des données (U-Th-Sm)/He sur apatite témoignent d'un refroidissement Oligocène enregistré uniquement dans la croûte supérieure de la bordure nord de la zone de faille Motagua. Nous tenterons ultérieurement de reproduire ce découplage vertical de la structure thermique par la modélisation de la formation d'un bassin transtensif et de circulation de fluides le long de la faille de Motagua. - This thesis focuses on the influence of the dynamic thermal structure of the upper crust (<10km) on the thermochronologic record of the exhumational and topographic history of magmatic continental arcs. Two mountain belts from the American Cordillera are studied: the North Cascades (USA) and the Motagua fault zone (Guatemala). I use a combined approach coupling apatite and zircon (U-Th-Sm}/He thermochronology and thermo- kinematic numerical modelling. This study highlights the temporal and spatial variability of the geothermal gradient and the importance to take into account the different geological processes that perturb the thermal structure of Cordilleran-type mountain belts (i.e. mountain belts related to oceanic subduction underneath a continent}.We integrate apatite and zircon (U-Th)/He data with numerical thermo-kinematic models to study the relative effects of magmatic and surface processes on the thermal evolution of the crust and cooling patterns in the Cenozoic North Cascades arc (Washington State, USA). Two age-elevation profiles that are located 7 km south of the well-studied Chiliiwack intrusions shows that spatial and temporal variability in geothermal gradients linked to magma emplacement can be contrained and separated from exhumation processes. During Chiliiwack batholith emplacement at -35-20 Ma, the geothermal gradient of the country rocks increased to a very high steady-state value (80-100°C/km), which is likely a function of magma flux and the distance from the magma source area. Including temporally varying geothermal gradients in the analysis allows quantifying the thermal perturbation around magmatic intrusions and retrieving a relatively simple denudation history from the data.The synthesis of new and previously published (U-Th)/He data reveals that denudation of the Northern Cascades is spatially and temporally constant at -100 m/Ma between ~32 and ~2 Ma, which likely reflects uplift due to magmatic crustal thickening since the initiation of the Cenozoic stage of the continental magmatic arc. In contrast, the humid flank of the North Cascades is affected by a ten-fold acceleration in exhumation rate at ~2 Ma, which we interpret as forced by the initiation of glaciations; around 3 km of crust have been eroded since that time. Glaciations have three distinct effects on the dynamics of this mountain range: (1) they increase erosion, exhumation and uplift rates where precipitation rates are sufficient to drive efficient glacial erosion; (2) they efficiently limit the elevation of the range; (3) they lead to widening of the humid flank and contraction of the arid flank of the belt.Drainage reorganizations constitute an important agent of landscape evolution that is often underestimated to the benefit of tectonic or climatic events. We propose a new method that integrates geomorphology, low-temperature thermochronometry (apatite and zircon {U-Th-Sm)/He), and 3D numerical thermal-kinematic modelling to detect and date drainage instability producing recent gorge incision, and apply this approach to the Skagit River Gorge, North Cascades.Two zircon (U-Th)/He age-elevation profiles sampled on both sides of the Motagua Fault Zone (MFZ), the boundary between the North American and the Caribbean plates, combined with published thermochronological data show that strike-slip displacement has juxtaposed the cold Maya block (s.s.) against the hot, arc derived, Chortis block (s.s ), producing different age patterns on both sides of the fault and short-wavelength lateral thermal exchange, resulting in recent heating of the cool side and cooling of the hot side of the MFZ.Finally, an apatite (U-Th-Sm)/He age-elevation profile records rapid cooling at -35 Ma localized only in the upper crust along the northern side of the Motagua fault zone. We will try to reproduce these data by modeling the thermal perturbation resulting from the formation of a transtensional basin and of fluid flow activity along a crustal- scale strike-slip fault.

Transcription factor CTF1 acts as a chromatin domain boundary that shields human telomeric genes from silencing.

Telomeres are associated with chromatin-mediated silencing of genes in their vicinity. However, how epigenetic markers mediate mammalian telomeric silencing and whether specific proteins may counteract this effect are not known. We evaluated the ability of CTF1, a DNA- and histone-binding transcription factor, to prevent transgene silencing at human telomeres. CTF1 was found to protect a gene from silencing when its DNA-binding sites were interposed between the gene and the telomeric extremity, while it did not affect a gene adjacent to the telomere. Protein fusions containing the CTF1 histone-binding domain displayed similar activities, while mutants impaired in their ability to interact with the histone did not. Chromatin immunoprecipitation indicated the propagation of a hypoacetylated histone structure to various extents depending on the telomere. The CTF1 fusion protein was found to recruit the H2A.Z histone variant at the telomeric locus and to restore high histone acetylation levels to the insulated telomeric transgene. Histone lysine trimethylations were also increased on the insulated transgene, indicating that these modifications may mediate expression rather than silencing at human telomeres. Overall, these results indicate that transcription factors can act to delimit chromatin domain boundaries at mammalian telomeres, thereby blocking the propagation of a silent chromatin structure.

Models that include supercoiling of topological domains reproduce several known features of interphase chromosomes.

Understanding the structure of interphase chromosomes is essential to elucidate regulatory mechanisms of gene expression. During recent years, high-throughput DNA sequencing expanded the power of chromosome conformation capture (3C) methods that provide information about reciprocal spatial proximity of chromosomal loci. Since 2012, it is known that entire chromatin in interphase chromosomes is organized into regions with strongly increased frequency of internal contacts. These regions, with the average size of ∼1 Mb, were named topological domains. More recent studies demonstrated presence of unconstrained supercoiling in interphase chromosomes. Using Brownian dynamics simulations, we show here that by including supercoiling into models of topological domains one can reproduce and thus provide possible explanations of several experimentally observed characteristics of interphase chromosomes, such as their complex contact maps.

Boundary between Soil and Saprolite in Alisols in the South of Brazil

Despite numerous studies conducted on the lower limit of soil and its contact with saprolite layers, a great deal of work is left to standardize identification and annotation of these variables in the field. In shallow soils, the appropriately noting these limits or contacts is essential for determining their behavior and potential use. The aims of this study were to identify and define the field contact and/or transition zone between soil and saprolite in profiles of an Alisol derived from fine sandstone and siltstone/claystone in subtropical southern Brazil and to subsequently validate the field observations through a multivariate analysis of laboratory analytical data. In the six Alisol profiles evaluated, the sequence of horizons found was A, Bt, C, and Cr, where C was considered part of the soil due to its pedogenetic structure, and Cr was considered saprolite due to its rock structure. The morphological properties that were determined in the field and that were different between the B and C horizons and the Cr layer were color, structure, texture, and fragments of saprolite. According to the test of means, the properties that support the inclusion of the C horizon as part of the soil are sand, clay, water-dispersible clay, silt/clay ratio, macroporosity, total porosity, resistance to penetration, cation exchange capacity, Fe extracted by DCB, Al, H+Al, and cation exchange capacity of clay. The properties that support the C horizon as a transition zone are silt, Ca, total organic C, and Fe extracted by ammonium oxalate. Discriminant analysis indicated differences among the three horizons evaluated.

Fatigue analysis of brackets with and without scallop for bridge and other deck structure using effective notch stress.

Due to functional requirement of a structural detail brackets with and without scallop are frequently used in bridges, decks, ships and offshore structure. Scallops are designed to serve as passage way for fluids, to reduce weld length and plate distortions. Moreover, scallops are used to avoid intersection of two or more welds for the fact that there is the presence of inventible inherent initial crack except for full penetrated weld and the formation of multi-axial stress state at the weld intersection. Welding all around the scallop corner increase the possibility of brittle fracture even for the case the bracket is not loaded by primary load. Avoiding of scallop will establish an initial crack in the corner if bracket is welded by fillet welds. If the two weld run pass had crossed, this would have given a 3D residual stress situation. Therefore the presences and absence of scallop necessitates the 3D FEA fatigue resistance of both types of brackets using effective notch stress approach ( ). FEMAP 10.1 with NX NASTRAN was used for the 3D FEA. The first and main objective of this research was to investigate and compare the fatigue resistance of brackets with and without scallop. The secondary goal was the fatigue design of scallops in case they cannot be avoided for some reason. The fatigue resistance for both types of brackets was determined based on approach using 1 mm fictitiously rounded radius based on IIW recommendation. Identical geometrical, boundary and loading conditions were used for the determination and comparison of fatigue resistance of both types of brackets using linear 3D FEA. Moreover the size effect of bracket length was also studied using 2D SHELL element FEA. In the case of brackets with scallop the flange plate weld toe at the corner of the scallop was found to exhibit the highest and made the flange plate weld toe critical for fatigue failure. Whereas weld root and weld toe at the weld intersections were the highly stressed location for brackets without scallop. Thus weld toe for brackets with scallop, and weld root and weld toe for brackets without scallop were found to be the critical area for fatigue failure. Employing identical parameters on both types of brackets, brackets without scallop had the highest except for full penetrated weld. Furthermore the fatigue resistance of brackets without scallop was highly affected by the lack of weld penetration length and it was found out that decreased as the weld penetration was increased. Despite the fact that the very presence of scallop reduces the stiffness and also same time induce stress concentration, based on the 3D FEA it is worth concluding that using scallop provided better fatigue resistance when both types of brackets were fillet welded. However brackets without scallop had the highest fatigue resistance when full penetration weld was used. This thesis also showed that weld toe for brackets with scallop was the only highly stressed area unlike brackets without scallop in which both weld toe and weld root were the critical locations for fatigue failure when different types of boundary conditions were used. Weld throat thickness, plate thickness, scallop radius, lack of weld penetration length, boundary condition and weld quality affected the fatigue resistance of both types of brackets. And as a result, bracket design procedure, especially welding quality and post weld treatment techniques significantly affect the fatigue resistance of both type of brackets.

Effect of different surface roughnesses on a turbulent boundary layer

The classical treatment of rough wall turbulent boundary layers consists in determining the effect the roughness has on the mean velocity profile. This effect is usually described in terms of the roughness function delta U+. The general implication is that different roughness geometries with the same delta U+ will have similar turbulence characteristics, at least at a sufficient distance from the roughness elements. Measurements over two different surface geometries (a mesh roughness and spanwise circular rods regularly spaced in the streamwise direction) with nominally the same delta U+ indicate significant differences in the Reynolds stresses, especially those involving the wall-normal velocity fluctuation, over the outer region. The differences are such that the Reynolds stress anisotropy is smaller over the mesh roughness than the rod roughness. The Reynolds stress anisotropy is largest for a smooth wall. The small-scale anisotropy and interniittency exhibit much smaller differences when the Taylor microscale Reynolds number and the Kolmogorov-normalized mean shear are nominally the same. There is nonetheless evidence that the small-scale structure over the three-dimensional mesh roughness conforms more closely with isotropy than that over the rod-roughened and smooth walls.

Structure et interactions de bulles d'espace-temps en relativité générale

Nous analysons des bulles d'espace-temps d'épaisseur finie en relativité générale. Les conditions d'énergie sont utilisées afin d'obtenir un ensemble de critères permettant de restreindre la structure du bord de la bulle. Dans le cas des bulles statiques et à symétrie sphérique, nous obtenons quatre inégalités différentielles équivalentes aux trois conditions d'énergie les plus communes. Nous montrons qu'elles sont équivalentes à un ensemble de deux inégalités différentielles simples lorsque le potentiel gravitationnel effectif a une forme particulière. Nous paramétrons alors l'espace-temps de manière à rendre la vérification de ces inégalités plus simple lorsqu'il sera question de bulles d'espace-temps. Nous traitons en particulier quatre formes de bulles, toutes caractérisées par un extérieur de type Schwarzschild de Sitter. Nous montrons que notre méthode donne les bons résultats lorsque la limite où l'épaisseur de la bulle tend vers zéro est prise. Nous terminons par un traitement succinct du problème d'une onde gravitationnelle se propageant dans un nuage de bulles d'espace-temps.

La structure de Jordan des matrices de transfert des modèles de boucles et la relation avec les hamiltoniens XXZ

Les modèles sur réseau comme ceux de la percolation, d’Ising et de Potts servent à décrire les transitions de phase en deux dimensions. La recherche de leur solution analytique passe par le calcul de la fonction de partition et la diagonalisation de matrices de transfert. Au point critique, ces modèles statistiques bidimensionnels sont invariants sous les transformations conformes et la construction de théories des champs conformes rationnelles, limites continues des modèles statistiques, permet un calcul de la fonction de partition au point critique. Plusieurs chercheurs pensent cependant que le paradigme des théories des champs conformes rationnelles peut être élargi pour inclure les modèles statistiques avec des matrices de transfert non diagonalisables. Ces modèles seraient alors décrits, dans la limite d’échelle, par des théories des champs logarithmiques et les représentations de l’algèbre de Virasoro intervenant dans la description des observables physiques seraient indécomposables. La matrice de transfert de boucles D_N(λ, u), un élément de l’algèbre de Temperley- Lieb, se manifeste dans les théories physiques à l’aide des représentations de connectivités ρ (link modules). L’espace vectoriel sur lequel agit cette représentation se décompose en secteurs étiquetés par un paramètre physique, le nombre d de défauts. L’action de cette représentation ne peut que diminuer ce nombre ou le laisser constant. La thèse est consacrée à l’identification de la structure de Jordan de D_N(λ, u) dans ces représentations. Le paramètre β = 2 cos λ = −(q + 1/q) fixe la théorie : β = 1 pour la percolation et √2 pour le modèle d’Ising, par exemple. Sur la géométrie du ruban, nous montrons que D_N(λ, u) possède les mêmes blocs de Jordan que F_N, son plus haut coefficient de Fourier. Nous étudions la non diagonalisabilité de F_N à l’aide des divergences de certaines composantes de ses vecteurs propres, qui apparaissent aux valeurs critiques de λ. Nous prouvons dans ρ(D_N(λ, u)) l’existence de cellules de Jordan intersectorielles, de rang 2 et couplant des secteurs d, d′ lorsque certaines contraintes sur λ, d, d′ et N sont satisfaites. Pour le modèle de polymères denses critique (β = 0) sur le ruban, les valeurs propres de ρ(D_N(λ, u)) étaient connues, mais les dégénérescences conjecturées. En construisant un isomorphisme entre les modules de connectivités et un sous-espace des modules de spins du modèle XXZ en q = i, nous prouvons cette conjecture. Nous montrons aussi que la restriction de l’hamiltonien de boucles à un secteur donné est diagonalisable et trouvons la forme de Jordan exacte de l’hamiltonien XX, non triviale pour N pair seulement. Enfin nous étudions la structure de Jordan de la matrice de transfert T_N(λ, ν) pour des conditions aux frontières périodiques. La matrice T_N(λ, ν) a des blocs de Jordan intrasectoriels et intersectoriels lorsque λ = πa/b, et a, b ∈ Z×. L’approche par F_N admet une généralisation qui permet de diagnostiquer des cellules intersectorielles dont le rang excède 2 dans certains cas et peut croître indéfiniment avec N. Pour les blocs de Jordan intrasectoriels, nous montrons que les représentations de connectivités sur le cylindre et celles du modèle XXZ sont isomorphes sauf pour certaines valeurs précises de q et du paramètre de torsion v. En utilisant le comportement de la transformation i_N^d dans un voisinage des valeurs critiques (q_c, v_c), nous construisons explicitement des vecteurs généralisés de Jordan de rang 2 et discutons l’existence de blocs de Jordan intrasectoriels de plus haut rang.

Dynamic characteristics of Atmospheric boundary Layer during different phases of Monsoon

Atmospheric Boundary layer (ABL) is the layer just above the earth surface and is influenced by the surface forcing within a short period of an hour or less. In this thesis, characteristics of the boundary layer over ocean, coastal and inland areas of the atmosphere, especially over the monsoon regime are thoroughly studied. The study of the coastal zone is important due to its high vulnerability mainly due to sea breeze circulation and associated changes in the atmospheric boundary layer. The major scientific problems addressed in this thesis are diurnal and seasonal variation of coastal meteorological properties, the characteristic difference in the ABL during active and weak monsoons, features of ABL over marine environment and the variation of the boundary layer structure over an inland station. The thesis describes the various features in the ABL associated with the active and weak monsoons and, the surface boundary layer properties associated with the active and weak epochs. The study provides knowledge on MABL and can be used as the estimated values of boundary layer parameters over the marine atmosphere and to know the values and variabilities of the ABL parameters such as surface wind, surface friction, drag coefficient, wind stress and wind stress curl.

PSWT Based Linear Predictive Coding and Development of a Parallel Multiple Subsequence Structure for DWT Computation

During 1990's the Wavelet Transform emerged as an important signal processing tool with potential applications in time-frequency analysis and non-stationary signal processing.Wavelets have gained popularity in broad range of disciplines like signal/image compression, medical diagnostics, boundary value problems, geophysical signal processing, statistical signal processing,pattern recognition,underwater acoustics etc.In 1993, G. Evangelista introduced the Pitch- synchronous Wavelet Transform, which is particularly suited for pseudo-periodic signal processing.The work presented in this thesis mainly concentrates on two interrelated topics in signal processing,viz. the Wavelet Transform based signal compression and the computation of Discrete Wavelet Transform. A new compression scheme is described in which the Pitch-Synchronous Wavelet Transform technique is combined with the popular linear Predictive Coding method for pseudo-periodic signal processing. Subsequently,A novel Parallel Multiple Subsequence structure is presented for the efficient computation of Wavelet Transform. Case studies also presented to highlight the potential applications.

Structural characterization and distribution of zebrafish germ plasm during interphase and mitosis

In zebrafish, germ cells are responsible for transmitting the genetic information from one generation to the next. During the first cleavages of zebrafish embryonic development, a specialized part of the cytoplasm known as germ plasm, is responsible of committing four blastomeres to become the progenitors of all germ cells in the forming embryo. Much is known about how the germ plasm is spatially distributed in early stages of primordial germ cell development, a process described to be dependant on microtubules and actin. However, little is known about how the material is inherited after it reorganizes into a perinuclear location, or how is the symmetrical distribution regulated in order to ensure proper inheritance of the material by both daughter cells. It is also not clear whether there is a controlled mechanism that regulates the number of granules inherited by the daughter cells, or whether it is a random process. We describe the distribution of germ plasm material from 4hpf to 24hpf in zebrafish primordial germ cells using Vasa protein as marker. Vasa positive material appears to be conglomerate into 3 to 4 big spherical structures at 4hpf. While development progresses, these big structures become smaller perinuclear granules that reach a total number of approximately 30 at 24hpf. We investigated how this transformation occurs and how the minus-end microtubule dependent motor protein Dynein plays a role in this process. Additionally, we describe specific colocalization of microtubules and perinuclear granules during interphase and more interestingly, during all different stages of cell division. We show that distribution of granules follow what seems to be a regulated distribution: during cells division, daughter cells inherit an equal number of granules. We propose that due to the permanent colocalization of microtubular structures with germinal granules during interphase and cell division, a coordinated mechanism between these structures may ensure proper distribution of the material among daughter cells. Furthermore, we show that exposure to the microtubule-depolymerizing drug nocodazole leads to disassembly of the germ cell nuclear lamin matrix, chromatin condensation, and fusion of granules to a big conglomerate, revealing dependence of granular distribution on microtubules and proper nuclear structure.

Image segmentation integrating colour, texture and boundary information

La tesis se centra en la Visión por Computador y, más concretamente, en la segmentación de imágenes, la cual es una de las etapas básicas en el análisis de imágenes y consiste en la división de la imagen en un conjunto de regiones visualmente distintas y uniformes considerando su intensidad, color o textura. Se propone una estrategia basada en el uso complementario de la información de región y de frontera durante el proceso de segmentación, integración que permite paliar algunos de los problemas básicos de la segmentación tradicional. La información de frontera permite inicialmente identificar el número de regiones presentes en la imagen y colocar en el interior de cada una de ellas una semilla, con el objetivo de modelar estadísticamente las características de las regiones y definir de esta forma la información de región. Esta información, conjuntamente con la información de frontera, es utilizada en la definición de una función de energía que expresa las propiedades requeridas a la segmentación deseada: uniformidad en el interior de las regiones y contraste con las regiones vecinas en los límites. Un conjunto de regiones activas inician entonces su crecimiento, compitiendo por los píxeles de la imagen, con el objetivo de optimizar la función de energía o, en otras palabras, encontrar la segmentación que mejor se adecua a los requerimientos exprsados en dicha función. Finalmente, todo esta proceso ha sido considerado en una estructura piramidal, lo que nos permite refinar progresivamente el resultado de la segmentación y mejorar su coste computacional. La estrategia ha sido extendida al problema de segmentación de texturas, lo que implica algunas consideraciones básicas como el modelaje de las regiones a partir de un conjunto de características de textura y la extracción de la información de frontera cuando la textura es presente en la imagen. Finalmente, se ha llevado a cabo la extensión a la segmentación de imágenes teniendo en cuenta las propiedades de color y textura. En este sentido, el uso conjunto de técnicas no-paramétricas de estimación de la función de densidad para la descripción del color, y de características textuales basadas en la matriz de co-ocurrencia, ha sido propuesto para modelar adecuadamente y de forma completa las regiones de la imagen. La propuesta ha sido evaluada de forma objetiva y comparada con distintas técnicas de integración utilizando imágenes sintéticas. Además, se han incluido experimentos con imágenes reales con resultados muy positivos.