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Introdução - A perda de estrutura dental devido às lesões cervicais não cariosas (LCNC) é um evento clínico recorrente e amplamente discutido na literatura. Os fatores etiológicos relacionados a esse tipo de lesão são: abrasão, erosão e abfração. Segundo vários autores, a particular disposição das estruturas dentinárias torna-se ainda mais interessante e curiosa quando se observa a dentina nas regiões com LCNC. Objetivos - A proposta deste trabalho foi estudar, comparativamente, em microscópio eletrônico de varredura (M.E.V.), a microestrutura dentinária de LCNC, a partir de dentes extraídos e armazenados sob condições controladas, considerando-se o método de preparação dos espécimes. Também se propôs a comparar o conteúdo intratubular da dentina com LCNC com sua contraparte lingual, sem lesão, e observar se a presença de estruturas intratubulares (EIT) tem relação com a ocorrência de hiperestesia dentinária (HD). Materiais e Métodos - Utilizaram-se as superfícies vestibulares e linguais homólogas de 14 dentes humanos, portadores de LCNC, extraídos por razões terapêuticas, e imediatamente fixados em Karnovsky. Os dentes foram divididos em 3 grupos, cada um deles subdivido em dois subgrupos, de acordo com o tratamento dos espécimes. Amostras da região da lesão e suas contrapartes linguais foram, então, pós-fixadas segundo protocolo apropriado para M.E.V. Tanto a superfície da lesão, quanto as estruturas dentinárias subjacentes foram estudadas obedecendo à seguinte seqüência: Grupo I - amostras da lesão seccionadas com disco, subdivididas em: Grupo IA - limpeza com ultra-som por 15 minutos; Grupo IB - limpeza com ultra-som por 15 minutos, seguida de condicionamento com H3PO4 a 5%, por 15 segundos. Grupos II - amostras fraturadas, subdivididas em: Grupo IIA - limpeza com ultra-som por 30 minutos; Grupo IIB - apenas condicionamento com H3PO4 a 5%, por 15 segundos; e Grupo III - amostras fraturadas e assim subdivididas: Grupo IIIA - controle, sem qualquer tipo de tratamento; Grupo IIIB - imersão dos espécimes em NaOCl a 2%, por 5 minutos. Os espécimes foram desidratados, submetidos a secagem em ponto crítico e metalizados com ouro para análise em M.E.V. Resultados - 1 - De maneira geral, todos os métodos interferem, em maior ou menor proporção, com a natureza dos espécimes. Na superfície da lesão, o método que melhor permitiu a visualização das estruturas dentinárias foi o ultra-som seguido do condicionamento ácido (Grupo IIA). Entretanto, é o procedimento que mais altera a estrutura da dentina. Na dentina abaixo da lesão, o método de observação mais apropriado foi o da simples fratura controle (Grupo IIIA). 2 - Tanto a dentina correspondente à região da lesão, quanto à da região não lesionada lingual, apresentaram conteúdos intratubulares semelhantes. 3 ? Não foram notadas diferenças estruturais nas lesões hipersensíveis em comparação com aquelas não-sensíveis. Conclusões - Os métodos de preparação dos espécimes tendem, em maior ou menor extensão, a alterar a natureza do que está sendo observado. Não se pode afirmar, à luz da metodologia empregada, que a existência de EIT é ocorrência particular da dentina em LCNC. Aparentemente, as EITs ocorrem tanto na dentina que apresenta HD como naquela não sensível. / Introduction - The lost of dental structure because of non carious cervical lesion is one of the most important clinical event discussed in dental literature. The most important etiologic factors related to this type of lesions are: abrasion, erosion and abfraction. According to many authors, this particular lost of dental structures became more interesting and curious when the dentin in regions with NCCL is study microscopically. Objectives - The purpose of this investigation was to study, comparatively, in scanning electronic microscopy (S.E.M.), the dental microstructure of NCCL and lesion-dentin dentin surfaces. The null hypothesis were the following: 1- the methods used for specimens preparations lead to different changes on dentin microstructures; 2- there is no difference between NCCL dentin microstructure and that from lesion-free dentin in the same tooth; 3- ITS can be found both in hypersensitive and non-sensitive dentin. Materials and Method - Dentin samples from buccal and lingual surfaces of 14 extracted human teeth, with NCCL on buccal surface, were used. After extraction the teeth were immediately fixed with Karnovsky. Teeth were than, divided in 2 subgroups, in accordance with specimen\'s treatment. Samples of region of lesion and their lingual counterpart were, then, post fixed according to appropriated protocol for SEM. The following groups were studied: Group I - specimens sectioned with diamond disc and subdivided in: Group IA- specimens cleaned in ultrasonic bath for 15 minutes; Group IB - cleaned in ultrasonic bath 15 minutes, followed by etching with 5%, H3PO4, for 15 seconds. Group II - samples were fractured along the lesions axis and subdivided in: Group IIA - ultrasonic bath for 30 minutes; Group IIB -etching with 5%, H3PO4, for 15 seconds. Group III - samples also fractured and subdivided in: Group IIIA (control) - without any kind of treatment; Group IIIB - immersion of the specimens in NaOCl to 2%, for 5 minutes. The specimens were dehydrated, critical point dried and coated with gold for in SEM analysis. Results - 1- On general, all of methods interfere, in larger or lesser proportion, with the nature of the specimens. On the surface of the lesions, the method that better allowed the visualization and recognition of dentine structures was the ultrasound followed by acid conditioning (Group IB). However, this was the procedure that introduced the greatest changes on dentine structures. In dentine under of the lesion, the best method of specimens preparation was the control group (Group IIIA). 2- Both buccal NCCL and lesion-free lingual surfaces presented similar ITS. 3- No structural differences were found in dentin from hypersensitive and non-sensitive lesions. Conclusions - The methods used for specimens tend to modify the characteristics and relationship of dentin structures. The hypothesis that ITS are specific from NCCL could not be confirmed under the light of method used. Apparently, the ITS can be found in both hypersensitive and non-hypersensitive dentin.

Introdução - A perda de estrutura dental devido às lesões cervicais não cariosas (LCNC) é um evento clínico recorrente e amplamente discutido na literatura. Os fatores etiológicos relacionados a esse tipo de lesão são: abrasão, erosão e abfração. Segundo vários autores, a particular disposição das estruturas dentinárias torna-se ainda mais interessante e curiosa quando se observa a dentina nas regiões com LCNC. Objetivos - A proposta deste trabalho foi estudar, comparativamente, em microscópio eletrônico de varredura (M.E.V.), a microestrutura dentinária de LCNC, a partir de dentes extraídos e armazenados sob condições controladas, considerando-se o método de preparação dos espécimes. Também se propôs a comparar o conteúdo intratubular da dentina com LCNC com sua contraparte lingual, sem lesão, e observar se a presença de estruturas intratubulares (EIT) tem relação com a ocorrência de hiperestesia dentinária (HD). Materiais e Métodos - Utilizaram-se as superfícies vestibulares e linguais homólogas de 14 dentes humanos, portadores de LCNC, extraídos por razões terapêuticas, e imediatamente fixados em Karnovsky. Os dentes foram divididos em 3 grupos, cada um deles subdivido em dois subgrupos, de acordo com o tratamento dos espécimes. Amostras da região da lesão e suas contrapartes linguais foram, então, pós-fixadas segundo protocolo apropriado para M.E.V. Tanto a superfície da lesão, quanto as estruturas dentinárias subjacentes foram estudadas obedecendo à seguinte seqüência: Grupo I - amostras da lesão seccionadas com disco, subdivididas em: Grupo IA - limpeza com ultra-som por 15 minutos; Grupo IB - limpeza com ultra-som por 15 minutos, seguida de condicionamento com H3PO4 a 5%, por 15 segundos. Grupos II - amostras fraturadas, subdivididas em: Grupo IIA - limpeza com ultra-som por 30 minutos; Grupo IIB - apenas condicionamento com H3PO4 a 5%, por 15 segundos; e Grupo III - amostras fraturadas e assim subdivididas: Grupo IIIA - controle, sem qualquer tipo de tratamento; Grupo IIIB - imersão dos espécimes em NaOCl a 2%, por 5 minutos. Os espécimes foram desidratados, submetidos a secagem em ponto crítico e metalizados com ouro para análise em M.E.V. Resultados - 1 - De maneira geral, todos os métodos interferem, em maior ou menor proporção, com a natureza dos espécimes. Na superfície da lesão, o método que melhor permitiu a visualização das estruturas dentinárias foi o ultra-som seguido do condicionamento ácido (Grupo IIA). Entretanto, é o procedimento que mais altera a estrutura da dentina. Na dentina abaixo da lesão, o método de observação mais apropriado foi o da simples fratura controle (Grupo IIIA). 2 - Tanto a dentina correspondente à região da lesão, quanto à da região não lesionada lingual, apresentaram conteúdos intratubulares semelhantes. 3 ? Não foram notadas diferenças estruturais nas lesões hipersensíveis em comparação com aquelas não-sensíveis. Conclusões - Os métodos de preparação dos espécimes tendem, em maior ou menor extensão, a alterar a natureza do que está sendo observado. Não se pode afirmar, à luz da metodologia empregada, que a existência de EIT é ocorrência particular da dentina em LCNC. Aparentemente, as EITs ocorrem tanto na dentina que apresenta HD como naquela não sensível. / Introduction - The lost of dental structure because of non carious cervical lesion is one of the most important clinical event discussed in dental literature. The most important etiologic factors related to this type of lesions are: abrasion, erosion and abfraction. According to many authors, this particular lost of dental structures became more interesting and curious when the dentin in regions with NCCL is study microscopically. Objectives - The purpose of this investigation was to study, comparatively, in scanning electronic microscopy (S.E.M.), the dental microstructure of NCCL and lesion-dentin dentin surfaces. The null hypothesis were the following: 1- the methods used for specimens preparations lead to different changes on dentin microstructures; 2- there is no difference between NCCL dentin microstructure and that from lesion-free dentin in the same tooth; 3- ITS can be found both in hypersensitive and non-sensitive dentin. Materials and Method - Dentin samples from buccal and lingual surfaces of 14 extracted human teeth, with NCCL on buccal surface, were used. After extraction the teeth were immediately fixed with Karnovsky. Teeth were than, divided in 2 subgroups, in accordance with specimen\'s treatment. Samples of region of lesion and their lingual counterpart were, then, post fixed according to appropriated protocol for SEM. The following groups were studied: Group I - specimens sectioned with diamond disc and subdivided in: Group IA- specimens cleaned in ultrasonic bath for 15 minutes; Group IB - cleaned in ultrasonic bath 15 minutes, followed by etching with 5%, H3PO4, for 15 seconds. Group II - samples were fractured along the lesions axis and subdivided in: Group IIA - ultrasonic bath for 30 minutes; Group IIB -etching with 5%, H3PO4, for 15 seconds. Group III - samples also fractured and subdivided in: Group IIIA (control) - without any kind of treatment; Group IIIB - immersion of the specimens in NaOCl to 2%, for 5 minutes. The specimens were dehydrated, critical point dried and coated with gold for in SEM analysis. Results - 1- On general, all of methods interfere, in larger or lesser proportion, with the nature of the specimens. On the surface of the lesions, the method that better allowed the visualization and recognition of dentine structures was the ultrasound followed by acid conditioning (Group IB). However, this was the procedure that introduced the greatest changes on dentine structures. In dentine under of the lesion, the best method of specimens preparation was the control group (Group IIIA). 2- Both buccal NCCL and lesion-free lingual surfaces presented similar ITS. 3- No structural differences were found in dentin from hypersensitive and non-sensitive lesions. Conclusions - The methods used for specimens tend to modify the characteristics and relationship of dentin structures. The hypothesis that ITS are specific from NCCL could not be confirmed under the light of method used. Apparently, the ITS can be found in both hypersensitive and non-hypersensitive dentin.

La douleur est un mécanisme essentiel pour la survie, mais les douleurs chroniques sont souvent invalidantes. Les douleurs de longue durée sont une des premières causes de consultation médicale, et affectent 20% de la population. Par contre, les traitements actuels sont souvent inefficaces au long cours, avec un ratio bénéfice/risque faible. Il est ainsi une priorité majeure la recherche des nouveaux agents thérapeutiques. Pour ce faire, la compréhension de réseaux impliqués dans la douleur est essentielle.Plusieurs études ont montré que Cav3.2, un canal calcique à bas seuil, pourrait être une cible analgésique prometteuse. En fait, Cav3.2 est exprimé tout le long de voies douloureuses, au niveau périphérique ainsi qu’au niveau central. Notamment, Cav3.2 est fortement exprimé dans la corne dorsale de la moelle épinière, qui constitue le premier relai d’intégration de l’information douloureuse. De plus, grâce à son activation aux potentiels proche du repos, Cav3.2 est bien placé pour réguler l’excitabilité neuronale et l’intégration synaptique. Au cours de cette thèse nous avons étudié la distribution de ce canal dans les interneurones de la lamina II de la corne dorsale. Nos résultats montrent que le canal est exprimé dans différentes types d’interneurones (~60%), majoritairement excitateurs, mais aussi inhibiteurs. De façon intéressante, tous les neurones PKCγ, impliqués dans l’allodynie mécanique, expriment le canal Cav3.2. Nous avons ensuite utilisé une approche virale pour marquer et étudier les propriétés électrophysiologiques des neurones exprimant Cav3.2, ainsi que pour réprimer de façon spécifiques le canal dans la partie lombaire de la moelle épinière. Nos résultats montrent que Cav3.2 participe dans les rebonds dépolarisants sous-liminaires, dans les propriétés des potentiels d’action et dans les profils d’activité électrique de ces neurones. Ainsi, Cav3.2 pourrait jouer un rôle important dans les états douloureux au niveau spinal, caractérisés par des changements dans l’excitabilité. / Acute pain is essential for survival, but chronic pain is often invalidating. Prolonged pain states are one of the first causes of medical consultation, and they are experienced by as much as 20% of the population. However, our current analgesics are not fully reliable in alleviating chronic pains, with low beneficial/risk ratio. Thus, finding new pharmacological agents are a primordial objective. A better understanding of pain networks is necessary.Several studies have suggested that the T-type calcium channel, Cav3.2, might be a promising target for analgesics. Indeed, Cav3.2 is expressed all along pain pathways, both within the peripheral nociceptive track and within the central nervous system. Interestingly, Cav3.2 is strongly express in the dorsal horn of the spinal cord, which is the first relay for pain processing. Furthermore, thanks to their activation near resting potentials, Cav3.2 channels fit well in regulating neuronal excitability and synaptic integration.In this thesis, we have analyzed Cav3.2 expression in dorsal horn interneurons. Our results show that Cav3.2 is expressed in different types of interneurons (~60%), mostly excitatory, but also inhibitory. Interestingly, all PKCγ neurons, involved in mechanical allodynia, expressed Cav3.2. We then used a viral approach to label and study electrophysiological properties of Cav3.2-expressing neurons, and also to specifically delete this channel in the lumbar spinal cord. Our results show that Cav3.2 is involved in subthreshold depolarizations, in the properties of the action potentials and in the firing patterns of these neurons. Thus, Cav3.2 might play an important role in pain states at the spinal cord level, characterized by neuronal excitability alterations.

Calcified teeth comprise the hardest vertebrate tissues (enamel and dentine) and they undoubtedly represent a key evolutionary advantage for their users. Nevertheless, teeth have been lost many times and they were often replaced by keratinised structures, such as rhamphoteca of birds or of turtle beaks, or labial tooth structures and scratching laminae of many aquatic vertebrates. This thesis is intended to analyse levels of similarities among keratinised teeth and structures of three phylogenetically distinct lineages of amniotes and to compare them to morphogenesis of true teeth. Detailed developmental and structural characteristics of oral apparatus possessing keratinous teeth of lampreys, Loricarid cathfishes and frog tadpoles were identified and distinguished. Keratinous structures of cathfishes include only horny projections from single cell surface (unculi), whereas in frog tadpoles both horny jaw sheet and unicellular labial teeth composed by cell columns were found; the lamprey oral apparatus consists of multicellular teeth and laminae instead. Disparate phenotypes of these keratinous structures can, however, developmentally derive from a general process of keratinization: morphological disparities arise by distinct gradual accretion of keratinous layers and collumns during development. My...

This thesis introduces lamina emergent mechanisms (LEMs) and presents components that can be used as building blocks to create LEMs capable of more complex motion. As the name suggests, lamina emergent mechanisms are fabricated out of planar materials (the lamina) but their motion is out of that plane (emergent). Lamina emergent mechanisms can provide benefits that include reduced manufacturing costs and minimal volume when in the planar state. The compact initial state of LEMs is beneficial in reducing shipping costs, especially in volume critical applications. LEMs also exhibit the potential benefits of compliant mechanisms, namely increased precision, reduced weight, reduced wear, and part count reduction. The LEM components presented in this thesis include flexible segments, fundamental mechanisms, and a new complaint joint, the lamina emergent torsional (LET) Joint. The flexible segments are developed through changes in geometry, boundary/loading conditions, and material. The fundamental mechanisms presented have motion similar to planar change-point four-bar and six-bar mechanisms, and spherical change-point mechanisms. The LET Joint is presented as a compliant joint suited for applications where large angular rotation is desired, but high off-axis stiffness is not as critical. The joint is introduced and the equations necessary for determining the force-deflection characteristics and stress are presented. Since the LET Joint can be fabricated from a single planar layer, it is well suited for macro and micro applications. Illustrative examples of the LET Joint are provided with devices fabricated from materials as diverse as steel, polypropylene, and polycrystalline silicon.

Multiple-layer Lamina Emergent Mechanisms (MLEMs) are mechanisms made from multiple sheets (lamina) of material with motion that emerges out of the fabrication plane. This study has shown that understanding how layers are used in existing products and in nature provides insight into how MLEMs can also use layers to achieve certain tasks. The multi-layered nature of MLEMs and the interactions between these layers are what enhance the capabilities of MLEMs and allow them to better meet design objectives. Layer separation is one objective for which MLEMs are well-suited. Layer separation can have a variety of applications and there are a number of different ways to design a MLEM to achieve this objective. Single-layer LEM and MLEM designs could greatly benefit from suitable actuation techniques; those that are consistent with the advantages of these mechanisms and could be incorporated into their design. This work presents shape memory alloys, piezoelectrics and dielectric elastomers as suitable ways of actuating LEMs and MLEMs. A number of novel MLEMs are presented throughout this thesis.

8 ABSTRACT Historically pig is allocated to a group of animals which use certain parts of their small intestine to acquire a fully developed primary B cell reperoire. Development of such primary repertoire is independent on the antigen presence and resembles the primary lymphopoietic activity of avian bursa of Fabricius. However, some findings concernig the pig's alignment in the above mentioned group suggest otherwise. This graduation thesis is focused on the investigtion of lymphocyte populations and subpopulations in the small intestine of germ-free and conventional piglets. The aim is to determine whether the percentage amounts of lymphocyte populations is dependent on the intestinal colonization. Using Flow Cytometry the significant differences between individual samples were assesed allowing us to conclude which parts of the small intestine could possibly be used for the development of B cell repertoire. Moreover, the status of isotype switching of B lymphocytes isolated from different intestinal parts was determined by the means of PCR analysis. Our data suggest that the small intestine colonization has a crucial role in development of all the main lymphocyte populations as well as some of their subpopulations. The greatest influence of colonization was observed concerning B lymphocytes and their...

Plants are exposed to various external stresses influencing physiology, anatomy, and morphology. Shape, geometry, and size of shoots and leaves are particularly affected. Among the latter, peltate leaves are not very common and so far, only few studies focused on their properties. In this case study, four Begonia species with different leaf shapes and petiole attachment points were analyzed regarding their leaf morphology, anatomy, and biomechanical properties. One to two plants per species were examined. In all four species, the petiole showed differently sized vascular bundles arranged in a peripheral ring and subepidermal collenchyma. These anatomical characteristics, low leaf dry mass, and low amount of lignified tissue in the petiole point toward turgor pressure as crucial for leaf stability. The petiole-lamina transition zone shows a different organization in leaves with a more central (peltate) and lateral petiole insertion. While in non-peltate leaves simple fiber branching is present, peltate leaves show a more complex reticulate fiber arrangement. Tensile and bending tests revealed similar structural Young’s moduli in all species for intercostal areas and venation, but differences in the petiole. The analysis of the leaves highlights the properties of petiole and the petiole-lamina transition zone that are needed to resist external stresses.

Sediment transport at the watershed scale in the Bluegrass Region of Kentucky is dominated by surface fine grained laminae, streambed, and streambank erosion; high instream sediment storage; and surface erosion processes. All these processes can be impacted by agricultural, urban, and suburban land-uses as well as hydrologic forcing. Understanding sediment transport processes at the watershed scale is a need for budgeting and controlling sediment pollution, and watershed modeling enables investigation of the cumulative effect of sediment processes and the parameters controlling these processes upon the entire sediment budget for a watershed. Sediment transport is being modeled by coupling the hydrologic model Hydrologic Simulations Program-FORTRAN (HSPF) with an in-house conceptually based hydraulic and sediment transport model. The total yield at the watershed outlet as well as the source fractions from surface fine grained lamina, streambed, and streambank sources; deposition; and biological generation within the streambed are predicted with the sediment transport model. Urbanization scenarios are then run on the calibrated model so as to predict the sediment budget for the South Elkhorn watershed for present and future conditions.