Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Os delfinídeos são os cetáceos mais ecologicamente diversos, ocorrendo numa ampla faixa de latitudes, em águas oceânicas e costeiras, incluindo regiões estuarinas e dulcícolas. O cenário taxonômico é especialmente confuso no gênero Tursiops, uma vez que grande parte das formas tem sido sinonimizadas na espécie Tursiopstruncatus. No entanto, estudos recentes sugerem que o gênero Tursiops seja polifilético. O golfinho-nariz-de-garrafa,T.truncatus, ocorre tanto em águas costeiras quanto oceânicas, em todas as regiões tropicais e temperadas. A espécie T. truncatus é tida como polimórfica e tal característica a torna alvo de acirradas discussões acerca do que são variações regionais ou diferentes entidades taxonômicas. O objetivo do presente estudo foi analisar a variabilidade morfológica de T. truncatus em distintas regiões oceânicas, buscando fornecer informações que permitam embasar os argumentos para futuras discussões taxonômicas que envolvem o gênero. Para isso, foi feita análise de Morfometria Geométrica em 2-D de crânios em vistas dorsal e lateral de espécimes que ocorrem nos oceanos Pacífico Norte Oriental, Atlântico Norte Ocidental, Atlântico Sul Ocidental, Atlântico Norte Oriental, Atlântico Sul Oriental e Índico. Foram encontradas diferenças significativas em todo o material analisado, incluindo diferenças entre exemplares reconhecidos como T. gephyreus e T. truncatus na costa Atlântica da América do Sul. As variações cranianas encontradas possuem relação com o tipo de ambiente em que os diferentes grupos ocorrem e podem estar relacionadas com a forma de forrageio, captura de presa e ao sistema de ecolocalização. Além disso, as variações na costa Atlântica da América do Sul podem ser explicadas pelo possível reconhecimento de duas espécies nessa região / Delphinids are the most ecologically diverse cetaceans, occurring in a wide range of latitudes in pelagic and coastal waters, including estuarine and freshwater regions. The taxonomy is especially confusing on genus Tursiops, since most forms has been synonymized in the specie Tursiopstruncatus. However, recent studies suggest that Tursiops is polyphyletic. The bottlenose dolphinT.truncatus habit coastal and pelagic waters in all tropical and temperate regions.Tursiopstruncatus is considered polymorphic and this characteristic makes the target of bitter taxonomic discussions about what are regional variations or different taxonomic entities. The objective was to analyze the cranial morphological variability of T. truncatus in different ocean regions to provide arguments for taxonomic discussions involving this genus. Two dimensional Geometric Morphometric analysiswas performed in skulls on dorsal and lateral views of specimens occurring in the Pacific North East, North West Atlantic, South West Atlantic, East North Atlantic, East South Atlantic and Indian oceans. There were significant differences between all groups, including differences between specimens recognizedasT. gephyreus and T. truncatus in the Atlantic coast of South America. The skull variationsare related to the type of environment in which different groups occur and may be relating to the foraging, prey capture and echolocation system. In addition, variations along the Atlantic coast of South America can be explained by the possible recognition of two species in this region

Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Os delfinídeos são os cetáceos mais ecologicamente diversos, ocorrendo numa ampla faixa de latitudes, em águas oceânicas e costeiras, incluindo regiões estuarinas e dulcícolas. O cenário taxonômico é especialmente confuso no gênero Tursiops, uma vez que grande parte das formas tem sido sinonimizadas na espécie Tursiopstruncatus. No entanto, estudos recentes sugerem que o gênero Tursiops seja polifilético. O golfinho-nariz-de-garrafa,T.truncatus, ocorre tanto em águas costeiras quanto oceânicas, em todas as regiões tropicais e temperadas. A espécie T. truncatus é tida como polimórfica e tal característica a torna alvo de acirradas discussões acerca do que são variações regionais ou diferentes entidades taxonômicas. O objetivo do presente estudo foi analisar a variabilidade morfológica de T. truncatus em distintas regiões oceânicas, buscando fornecer informações que permitam embasar os argumentos para futuras discussões taxonômicas que envolvem o gênero. Para isso, foi feita análise de Morfometria Geométrica em 2-D de crânios em vistas dorsal e lateral de espécimes que ocorrem nos oceanos Pacífico Norte Oriental, Atlântico Norte Ocidental, Atlântico Sul Ocidental, Atlântico Norte Oriental, Atlântico Sul Oriental e Índico. Foram encontradas diferenças significativas em todo o material analisado, incluindo diferenças entre exemplares reconhecidos como T. gephyreus e T. truncatus na costa Atlântica da América do Sul. As variações cranianas encontradas possuem relação com o tipo de ambiente em que os diferentes grupos ocorrem e podem estar relacionadas com a forma de forrageio, captura de presa e ao sistema de ecolocalização. Além disso, as variações na costa Atlântica da América do Sul podem ser explicadas pelo possível reconhecimento de duas espécies nessa região / Delphinids are the most ecologically diverse cetaceans, occurring in a wide range of latitudes in pelagic and coastal waters, including estuarine and freshwater regions. The taxonomy is especially confusing on genus Tursiops, since most forms has been synonymized in the specie Tursiopstruncatus. However, recent studies suggest that Tursiops is polyphyletic. The bottlenose dolphinT.truncatus habit coastal and pelagic waters in all tropical and temperate regions.Tursiopstruncatus is considered polymorphic and this characteristic makes the target of bitter taxonomic discussions about what are regional variations or different taxonomic entities. The objective was to analyze the cranial morphological variability of T. truncatus in different ocean regions to provide arguments for taxonomic discussions involving this genus. Two dimensional Geometric Morphometric analysiswas performed in skulls on dorsal and lateral views of specimens occurring in the Pacific North East, North West Atlantic, South West Atlantic, East North Atlantic, East South Atlantic and Indian oceans. There were significant differences between all groups, including differences between specimens recognizedasT. gephyreus and T. truncatus in the Atlantic coast of South America. The skull variationsare related to the type of environment in which different groups occur and may be relating to the foraging, prey capture and echolocation system. In addition, variations along the Atlantic coast of South America can be explained by the possible recognition of two species in this region

Dentro da biologia evolutiva, uma das questões centrais é a compreensão de como os processos evolutivos, em particular a seleção e o acaso (deriva genética) moldaram a diversidade obervada nos organismos. Dentro deste contexto, a utilização de abordagens como a integração morfológica e a genética quantitativa nos fornecem poderosas ferramentas. Enquanto a primeira descreve os padrões de relação entre caracteres e testa hipóteses sobre as relações de desenvolvimento e/ou funções subjacentes, a segunda possui ferramentas para investigar as forças que podem ter gerado os fenótipos atuais. Utilizando como modelo de estudo os gêneros de marsupiais da Ordem Didelphimorphia, analisei a evolução morfológica craniana unindo estas duas linhas de pesquisa. Central a genética quantitativa está a matriz de covariância genética (G) que descreve a porção da variância que é efetivamente herdada (no sentido de transmissão de valor entre gerações) e conseqüentemente serve como substrato à seleção. Apesar de ter sido desenvolvida inicialmente para estudos em escala microevolutiva, o arsenal matemático da genética quantitativa pode ser estendida a escalas macroevolutivas caso a matriz G permaneça relativamente estável. Entretanto, como a estimativa de uma matriz G requer um número grande de espécimes aparentados e com genealogia conhecida, utilizei ao longo deste trabalho sua correspondente fenotípica (P). Desta forma, no primeiro capítulo demonstrei a similaridade das matrizes de covariância e correlação P dos marsupiais Didelphimorphia. Em contrapartida, as magnitudes de integração morfológica, que medem o grau de associação entre os caracteres, se mostraram mais variáveis dentro do grupo. A história evolutiva (filogenia) e as distâncias morfológicas entre os gêneros não parecem ter tido influência sobre os padrões nem as magnitudes, já que não se correlacionaram significativamente a eles. No segundo capítulo, estendi estas análises para os gêneros ao longo do seu desenvolvimento e novamente os padrões se mostraram semelhantes, tanto quando comparei a ontogenia de um único gênero, quando analisei diferentes gêneros para diferentes classes etárias. Em contrapartida, as magnitudes de integração se mostraram mais variáveis, com uma tendência a diminuírem com o aumento da idade. Por conta destes resultados, comparei as matrizes de correlação empíricas com matrizes teóricas que expressam hipóteses de modularidade baseadas no desenvolvimento/função compartilhada pelas regiões do crânio. Testei para similaridade das duas regiões principais (face e neurocrânio), cinco sub-regiões (base e abóboda craniana, oral, nasal e zigomático), uma matriz 198 de integração somática (caracteres neurais vs. faciais) e uma de integração total, concatenando as cinco sub-regiões. Foram encontradas somente correlações significativas para as regiões da face e suas sub-regiões nasal e oral. Apesar das variações nas magnitudes de integração observadas nestes dois primeiros capítulos, as respostas evolutivas obtidas para estes gêneros foram todas muito similares. Este resultado, associado a constatação de que grande parte da variação entre os espécimes é devida ao tamanho, ou variações de forma associadas a ele (alometria), me levaram a analisar as conseqüências evolutivas associadas a estas variações alométricas. Assim, no terceiro capítulo analisei as direções de resposta à seleção simulada antes e após a remoção do tamanho. Variações alométricas exercem forte influência sobre estes crânios tornando-os muito integrados morfologicamente, ou seja, a percepção dos módulos que os compõem é muito difícil (baixa modularidade). Isto se reflete em uma maior variação ao longo da linha de menor resistência evolutiva (que resume o eixo de maior variação disponível entre os espécimes), já que os módulos não podem responder a seleção de forma independente (alta integração morfológica). Em marsupiais, esta linha é está alinhada na direção de variações de tamanho. Desta forma, não importa em que direção a seleção esteja atuando, as respostas evolutivas serão usualmente na direção desta linha de menor resistência e conseqüentemente de tamanho. Uma vez que o tamanho é removido, as magnitudes de integração diminuem e a modularidade do crânio aumenta. Por conseguinte, ele passa a ser capaz de responder à seleção em uma gama maior de direções porque aumenta também a relativa independência destes módulos em relação aos demais. Como estes resultados indicam que variações entre os Didelphimorphia são direcionadas a variações no tamanho, devido justamente às restrições impostas pela linha de menor resistência evolutiva, analisei as trajetórias ontegenéticas de dois gêneros irmãos, Didelphis e Philander, separando os efeitos da forma e tamanho nesta diversificação. Para tanto empreguei análises de morfometria tradicional, morfométrica e análises de coeficientes alométricos que apontaram para uma maior diferenciação no tamanho entre eles. Apesar da similaridade entre as trajetórias ontogenéticas destes dois gêneros, espécimes de Didelphis nascem maiores e crescem por mais tempo, como indicado pela sua trajetória mais longa. / One of the central goals in evolutionary biology is related to how evolutionary processes, mainly natural selection and genetic drift, shaped living organisms. The combined use of Morphologial Integration and Quantitative Genetics give us powerful tools to accomplish this goal. Morphological Integration is concerned of how characters are related, as well as their underlying genetics/developmental relationship, while Quantitative Genetics have methodologies designed to explore the phenotypic forces underlying diversity among organisms. Using Didelphimorphia marsupials as a study group, I combined these two approaches to study its morphologic cranial diversification. In Quantitative Genetics, the genetic additive covariance matrix (G) resume the genetic variation underlying resemblance among relatives, which is the portion of the variance responsive to selection. Initially developed to microevolutionary scale studies, it can be extended to a macroevolutionary scale if it remains relatively similar in that time scale. However, as G matrix estimations require a huge number of related specimens with known genealogy, I used its phenotypic (P) counterpart which was more easily obtained. In the first chapter I showed high similarities among Didelphimorphia marsupials covariance and correlation P matrices. On the other hand, integration magnitudes (which measure the average correlation among traits) vary among taxa. Neither phylogeny nor morphologic distances showed any association with the similarity in patterns and magnitudes of integration. In the second chapter, I did these same analyses, but throughout genera ontogeny. Again, there was a high similarity among taxa in patterns of integration, both when I analyzed the ontogeny for each genus separately or against each other (at different age class). Morphological integration magnitudes showed the same variation obtained for adults, with a tendency to decrease at older ages. Taking these results into account, I compared the phenotypic correlation matrices to theoretical matrices, based on hypotheses of shared developmental and functional units. I searched for modularity in the two main skull regions (face and neurocranium) and five sub regions (cranium base and vault, face, nasal, and oral). I also looked for modularity concerning somatic development (Neurocranium vs. Face) and total modularity, as the 200 summation of the five sub-regions. Only Face and its sub-regions nasal and oral, showed significant correlations to the phenotypic genera matrices. Despite integration magnitude differences, all evolutionary responses produced by taxa were highly similar. These results, combined with a huge size variation (or size related variation - allometry) across taxa, lead me to search for the evolutionary consequences due to size variation. In the third chapter, I compared evolutionary response directions produce by each genera matrices before and after size removal under a random selection simulation. Allometry strongly affect these skulls, turning them into highly integrated structures with lower modularity (skull modules are not easily distinguished). Because of this, modules cannot evolve relatively independent of other modules and evolutionary responses will strongly affect the whole cranium. This is related to the variation along the lines of least evolutionary resistance. This line is the multivariate direction of greatest genetic or phenotypic variation (the combination of a suite of traits that displays the maximum within-population variance). In Didelphimorphia marsupials, this line is aligned with size variation and regardless the selection direction, evolutionary change is usually aligned to this least resistance line. The removal of size variation diminish the magnitude of integration while increases modularity. Consequently, skulls become able to respond to selection in more directions as modules become relatively more independent of each other. In the last chapter I compared size and shape differences between ontogenetic trajectories of two sister genera Didelphis and Philander. Using traditional and geometric morphometric analysis plus allometric coefficient analysis, I could show that bigger differences between them are size related. Despite similarities, ontogenetic trajectory in Didelphis is longer, leading to bigger specimens.

A expansão craniana com o uso de molas tem demonstrado eficácia no tratamento das anormalidades craniofaciais, tais como as craniossinostoses. A ação expansora exercida pelas molas tem sido observada tanto quando colocadas entre as margens parietais dos ossos do crânio, como quando lateralmente à sutura sagital, principalmente nas escafocefalias. No presente estudo foi criado um modelo experimental com coelhos, e feita uma avaliação descritiva do comportamento da calota craniana e das suturas sob ação de molas. Foram utilizados 13 coelhos Nova Zelândia com quatro semanas de vida, divididos em quatro grupos: grupo I, foram implantados no crânio marcadores de amálgama para controle; no grupo II, marcadores de amálgama e osteotomia da sutura sagital; no grupo III, marcadores de amálgama, osteotomia da sutura sagital e colocação de uma mola expansora na região interparietal e, no grupo IV, marcadores de amálgama, craniotomia parassagital linear com colocação da mola. Os animais foram sacrificados com duas, quatro, oito e doze semanas. Foi realizado controle radiológico com avaliação do afastamento dos marcadores de amálgama, da variação dos ângulos cefalométricos e das medidas da base do crânio, bem como um estudo histopatológico da região de colocação das molas. Nos grupos com o uso de molas a separação dos bordos da craniotomia foi maior do que naqueles sem a utilização de mola. Houve ossificação em todos os grupos, com maior rapidez no grupo II. O crescimento ósseo deu-se a partir dos bordos e da profundidade. Não foram observadas diferenças significativas no padrão histopatológico da regeneração óssea entre os grupos com colocação de mola na região interparietal e parassagital. O modelo experimental com coelhos se mostrou adequado às análises propostas pelo estudo. Concluiu-se que houve osteogênese por distração nos grupos III e IV e que apresentaram uma expansão craniana semelhantes. / Spring-mediated skull expansion has proven to be effective in the treatment of craniofacial abnormalities, such as craniosynostosis. The use of springs in cranial expansion has been studied both in the sagittal and in parasagittal regions, especially in scaphocephaly. A rabbit model was used in the present study to analyze the effects of springs on the cranial vault and sutures. Thirteen 4-week-old New Zealand rabbits were used and divided into 4 groups: group I, amalgam markers were used as control; in group II, amalgam markers and osteotomy of the sagittal suture; in group III, amalgam markers and osteotomy of the sagittal suture with implant of an expansible spring in the interparietal region and in group IV, amalgam markers and linear parasagittal craniectomy with springs. Animals were sacrificed after 2, 4 and 12 weeks. Radiological control with assessment of the amalgam markers, variation of cephalometric angles and cranial base measurements, as well as the histopathological analysis of the spring implant area were carried out. In the groups using springs distraction of the craniectomy borders was greater than in those that did not use springs. Ossification was observed in all of the groups and was faster in group II. Bone growth started from the borders and depth. There were no significant differences in the histopathological pattern of bone regeneration between the groups with spring implant in the interparietal and parasagittal region. The rabbit model proved to be adequate for the analysis proposed by the study. It was concluded that there was osteogenesis caused by distraction in groups III and IV, with similar skull expansion rates.

Ce travail de thèse vise à mieux décrire les mécanismes de lésions de la tête humaine en situation de choc en optimisant le modèle par éléments finis de la tête humaine de Strasbourg (SUFEHM) en termes de modélisation mécanique du crâne et du cerveau grâce à de nouvelles données expérimentales et de techniques récentes d’imagerie médicales. Une première étape a consisté à améliorer la loi de comportement de la boîte crânienne, valider son comportement en regards d’éléments expérimentaux sur cadavres et proposer un MEF capable de reproduire fidèlement la fracture crânienne. La deuxième partie consiste en la prise en compte pour la première fois de l’anisotropie dans les simulations par EF d’accidents réels en utilisant l’Imagerie du Tenseur de Diffusion. Après implémentation, une phase de validation a été entreprise afin de démontrer l’apport de l’anisotropie de la matière cérébrale dans un MEF. Enfin 125 accidents réels ont été reproduits avec le SUFEHM ainsi amélioré. Une étude statistique sur les paramètres mécaniques calculés a permis de proposer des limites de tolérances en termes de fracture crânienne et de lésions neurologiques en s’intéressant tout particulièrement à l’élongation axonale maximale admissible, nouvelle métrique proposée. / The principal objective of this study is to enhance the existing finite element head model. A composite material model for skull, taking into account damage is implemented in the Strasbourg University Finite Element Head Model in order to enhance the existing skull mechanical constitutive law. The skull behavior is validated in terms of fracture patterns and contact forces by reconstructing 15 experimental cases in collaboration with Medical College of Wisconsin. The new skull model is capable of reproducing skull fracture precisely. The composite skull model is validated not only for maximum forces, but also for lateral impact against actual force time curves from PMHS for the first time. This study also proposes the implementation of fractional anisotropy and axonal fiber orientation from Diffusion Tensor Imaging of 12 healthy patients into an existing human FE head model to develop a more realistic brain model with advanced constitutive laws. Further, the brain behavior was validated in terms of brain strain against experimental data. A reasonable agreement was observed between the simulation and experimental data. Results showed the feasibility of integrating axonal direction information into FE analysis and established the context of computation of axonal elongation in case of head trauma. A total 125 reconstructions were done by using the new advanced FEHM and the axonal strain was found to be the pertinent parameter to predict DAI.

Apparue à la fin des années 1990, la tractographie utilise le signal diffusion de l’imagerie par résonance magnétique (IRM) pour détecter l’orientation préférentielle des molécules d’eau et reconstruire l’architecture des tissus biologiques, notamment celle des fibres blanches intra cérébrales. Cette technique a suscité l’engouement de la communauté scientifique en permettant, pour la première fois, l’étude in vivo non invasive des structures anatomiques, et en particulier celle du cerveau. Néanmoins, la description de la trajectoire des fibres blanches reste imprécise dans les zones de croisement de fibres et pour les faisceaux de petite taille comme les nerfs crâniens. De multiples méthodes sont développées aux différents étapes d’acquisition et de post-traitement afin d’améliorer la résolution spatiale et angulaire et augmenter la précision de la reconstruction des fibres. En pratique clinique, la détection de la trajectoire des nerfs crâniens déplacés au contact des tumeurs de la base du crâne pourrait apporter une aide significative dans la stratégie chirurgicale et améliorer le résultat fonctionnel pour les patients.Après avoir rappelé les notions fondamentales nécessaires à la compréhension de chaque étape de la tractographie, nous en présentons l’ « état de l’art » dans le cas particulier des nerfs crâniens. A partir de 21 études de la littérature scientifique, nous détaillons tous les paramètres d’acquisition et de tracking, les algorithmes de reconstruction, le design des régions d’intérêt et le filtrage. Puis, nous développons notre propre pipeline de tractographie et montrons son impact sur la prise en charge chirurgicale à travers une série de 62 cas de tumeurs variées de la base du crâne et 2 vignettes cliniques illustratives. Enfin, nous proposons une nouvelle approche, la full-tractography, avec une utilisation potentielle en routine clinique, notamment lors du planning pré-chirurgical / Tractography is a recent imaging tool that used the diffusion signal from the magnetic resonance imaging (MRI) to detect the preferential orientation of water molecules within the tissues and particularly along white fibers of the brain. This technique has caught the attention of the scientific community describing non-invasively the in vivo white matter architecture. Nonetheless, its application to fiber crossing areas or to small-scale structures, such as cranial nerves, remains inaccurate. New methods are being developed for both the acquisition and post-processing steps to provide a higher angular and spatial resolution imaging, and improve the reconstruction of fibers. In the clinical setting, the detection of the trajectory of the cranial nerves displaced by skull base tumors could be a relevant asset for the surgical strategy and the functional outcome. After reminding the basics to understand each step involved in tractography, we present the current state-of-the-art for application to cranial nerves. From 21 selected studies, we report all parameters of acquisition and tracking, algorithms of reconstruction, the design of the regions of interest, and filtering. We then develop our tractography pipeline and show its value for the surgical management through a 62 case series of various skull base tumors and two clinical images. Finally, we propose a new full-tractography approach that could be implemented in routine, notably for presurgical planning

Background and Aim: „CI-Wizard“ is a new, three-dimensional software planning tool for cochlear implant surgery with manual and semi-automatic algorithms to visualize anatomical risk structures of the lateral skull base preoperatively. Primary endpoints of the study represented the evaluation of the CI Wizards usability, accuracy, subjectively perceived and objectively measured time in clinical practice. Materials and Methods: In a period from January 2014 to March 2015, n=36 participants were included in this study. These members were divided into three groups of equal number (n=12), but different level of experience. Senior doctors and consultants (group 1), residents (group 2) and medical students (group 3) segmented twelve different CT-scan data sets of the CI Wizard (four per participant). In total, n=144 data sets were collected. The usability of the CI Wizard was measured by the given questionnaire with an interval rating scale. The Jaccard coefficient (JT) was used to evaluate the accuracy of the anatomical structures segmented. The subjectively-perceived time was measured with an interval rating scale in the questionnaire and was compared with the objectively mean measured time (time interact). Results: Across all three groups, the usability of the CI Wizard has been assessed between 1 ('very good') and 2 ('with small defects'). Subjectively, the time was stated as 'appropriate' by questionnaire. Objective measurements of the required duration revealed averages of n=9.8 minutes for creating a target view. Concerning the accuracy, semi automatic anatomical structures such as the external acoustic canal (JT=0.90), the tympanic cavity (JT=0.87), the ossicles (JT=0.63), the cochlea (JT=0.66) and the semicircular canals (JT=0.61) reached high Jaccard values, which describes a great match of the segmented structures between the partcipants and the gold standard. Facial nerve (JT=0.39) and round window (JT=0.37) reached lower Jaccard values. Very little overlap tendency was found for the chorda tympani (JT=0.11). Conclusion: This software program represents a further important step in the development of pre-operative planning tools in cochlear implant surgery. The study revealed a high level of satisfaction in the usability. The subjectively required time was considered as „appropriate“ and the objectively mean measured time was n=9.8 minutes short enough, so that a clinical application seems realistic. Particularly for semi-automatically segmented structures, it represented a good accuracy. For purely manual segmented structures, further improvements are desirable. Finally, this program also provides a good learning tool for medical students and residents to become familiar with the anatomy of the lateral skull base.:1 Einführung 1 1.1 Cochlea Implantate (CI) 1 1.2 Bildverarbeitungsprogamme zur präoperativen Planung eines Cochlea Implantates 2 1.2.1 Segmentierung in der Medizin 2 1.2.2 CI-Wizard 4 1.3 Fragebogen 6 1.4 Zielsetzung 6 1.5 Zusammenfassung der Ergebnisse 7 2 Publikation 8 3 Zusammenfassung 19 4 Literaturverzeichnis 23 Anlagen 26 Beitrag der Promovendin bei geteilter Erstautorenschaft 34 Erklärung über die eigenständige Abfassung der Arbeit 35 Lebenslauf 36 Danksagung 37

Bones are essential vertebrate structures that arise during embryonic development from mesenchymal condensations. In the skull vault, bones arise from condensations above the eye, which later expand to cover the brain. Contrarily to most bones in the axial skeleton, which develop through an intermediate cartilage template, the skull vault develops from the direct differentiation of mesenchymal progenitors into bone cells, or osteoblasts. The molecular details of this process have however remained elusive, since systems allowing the dynamical observation of cells while they undergobone differentiation have been lacking. Recent data in the Tabler lab, acquired by imaging skull caps ex vivo, has uncovered that differentiation takes place during expansion stages, and that a progressive wave of differentiation underlies the presence of intermediate states between progenitors and osteoblasts. In this thesis, we focused on the cranial mesenchyme to study the differentiation trajectory of skull bone progenitors at single cell resolution. By harnessing single cell RNA-Sequencing, we elucidated the heterogeneity of cells in the cranial mesenchyme, describing in molecular terms meningeal, dermal, osteoblastic and progenitor populations. By modelling dynamics from single cell RNA-Seq data, we inferred that a population expressing intermediate levels of cartilage specific genes, such as Col2a1, underlies differentiation towards dermal, meningeal and bone fate. Using single cell resolution in situ RNA hybridisation, we mapped the anatomical location of progenitors in a layer between the meninges and the bone. To better understand the relationship between the phenotype of these progenitors and differentiated cartilage, we examined the presence of cartilage-like ECM in the tissue. Finally, we asked whether similar progenitors may be present in other intramembranous bones outside the skull by re-applying these tools on the clavicle. Taken together, our data lead us to hypothesise a mechanism for differentiation of the cranial mesenchyme, which can explain previous phenotypes reported in the literature and supports a role for cartilage-like cells in intramembranous ossification.

In order to understand the relationship between feeding behavior and the evolution of mammalian skull form, it is essential to evaluate the impact of bite force over large regions of skull. There are about 1,100 bat species worldwide, which represent about 20% of all classified mammal species. Hence, a study in the evolution of bat skull form may provide general understanding of the overall evolution of skull form in mammals. These biomechanical studies are generally performed by first building solid Finite Element (FE) models of skull from micro CT scans. This process of building FE models from micro CT scans is both tedious and time consuming. Therefore a new approach is developed in this research project to build these FE models quickly and efficiently. I have used SolidWorks to build a parameterized, three dimensional surface CAD model of a skull of the short-tailed fruit bat, Carollia perspicillata, by using coordinate data from an STL model of the species. The overall shape of this model closely resembled that of solid model of C. perspiciallata constructed from micro CT scans. Finite element analyses of the solid and surface models yielded comparable results in terms of magnitude and distribution of von Mises stress and mechanical advantage. Using this parametric surface model, the FE plate or shell element models of different bat species were generated by varying two parameters, palate length and palate width. Parametric analyses were performed on these FE plate models of skulls and response surfaces of performance criteria: von Mises stress, strain energy and mechanical advantage were generated by varying the input parameters. After generating response surfaces, species of bats from the morphologically diverse family of New World leaf-nosed bats (Family Phyllostomidae) were overlain on these response surfaces to determine which portions of the performance design space (palate length X width) are and are not occupied. These plots serve as a foundation for understanding the affect of different performance criteria on the evolution of bat skull form.

In this dissertation, I examine the Sonoran Desert trilogy by Kathleen Alcalá through the lens of post-oppositional theory as developed by AnaLouise Keating. Moving beyond the use of post-oppositional theory to analyze non-fiction works, I apply this theory instead to the fiction of Kathleen Alcalá—whose work appears in such anthologies as The Norton Anthology of Latino Literature. Alcalá, though well published, is underrepresented in contemporary literary criticism, as can be seen by the only eight entries under her name in the MLA International Bibliography. Therefore, I have chosen her most significant fiction work, her trilogy about the Sonoran Desert, as the perfect text upon which to map post-oppositional theory. Through analysis of her three novels, I show that her work is an ideal example of post-oppositionality in action and that her characters act as post-oppositional revolutionaries and prophets within the pages of the text. The first chapter outlines the parameters of the project. In Chapter 2, I argue that post-oppositionality can be seen in Alcalá through gender bending, looking at the characters of Membrillo and Manzana, Corey, and Rosalinda. In Chapter 3, I argue that the characters of Estela, La Señorita, and Magdalena are enacting post-oppositionality through their transcendence of traditional women's roles in sexuality. In Chapter 4, I argue that the female characters of the novels act as revolutionaries through their political and social agency—reaching out to other characters through such work as educating and writing. In Chapters 5 and 6, I feature my interviews with Alcalá and Keating, who were generous enough to speak with me over Zoom during lockdown. Finally, in the conclusion chapter, Chapter 7, I examine how post-oppositionality in the novels prepares the reader for post-oppositional action in reality. Throughout all of these chapters, I rely on other theories and historiographies such as gender theory (Judith Butler, Foucault, West and Zimmerman, etc.), the history of women's sexuality, and the roles of women in nineteenth century Mexico (looking especially at the works of Nancy LaGreca and Anna Macías).