El presente trabajo de tesis estudia diferentes mutantes zeste1 y el efecto que esta mutación tiene sobre la regulación de la transcripción en diversos genes. En primer lugar se estudiaron los machos de las cepas M115 y RM115 cuya característica principal, además de ser portadores de la mutación z1, es la inserción de un elemento FB-NOF en el tercer intrón del gen CG32795, que forma con white un par de genes head-to-tail. Para estudiar el efecto de la inserción de FB-NOF en un entorno z1, fue necesario en primer lugar caracterizar el gen CG32795, a nivel de secuencia del mRNA y de predicción de la posible función de la proteína codificada. Se encontraron diversas variantes de splicing, tanto para su extremo 5' como para el 3', parecidas pero diferentes de las variantes predichas anteriormente. Conociendo más en profundidad este gen, nos propusimos estudiar los posibles efectos de la inserción de FB-NOF respecto a la expresión de los genes w y CG32795. Los resultados nos llevaron a un estudio más detallado de la expresión de w en diferentes partes del cuerpo, teniendo en cuenta la especificidad de tejido que la interacción zeste-white presenta. Así, demostramos que el gen w no se ve afectado por la inserción de FB-NOF en su extremo 3' y que las diferencias de expresión observadas son debidas a la duplicación del Zeste Binding Site de w en un entorno z1. Sin embargo, la inserción de FB-NOF en el tercer intrón de CG32795 si modifica la expresión de este gen. La inserción no es sólo responsable de la alteración de la expresión de CG32795 sino también la reordenación que se produce en la cepa RM115 eliminando la copia de w original y dejando la que se duplicó en M115.La segunda parte de esta tesis estudia las hembras mutantes z1. Analizamos la expresión de dos genes con un ZBS (w y dpp) y observamos como se reduce la expresión de dichos genes en las hembras z1. Además, estudiando la expresión del gen CG32795 constatamos que el efecto de la interacción zeste-white es local, sin afectar a CG32795 aunque su extremo 5' se encuentra a tan solo 700bp del extremo 3' del gen w. La reducción de la expresión podría ser debida a alteraciones en la estructura de la cromatina, puesto que los agregados de Zeste en los ZBS son los responsables de reclutar el complejo remodelador de la cromatina BRM, facilitando así la transcripción. Mediante un ensayo de nucleasa micrococal detectamos algunas modificaciones en el posicionamiento de nucleosomas en los ZBS de w y dpp, siendo el posicionamiento en las hembras z1 más estricto. Si el complejo BRM es el responsable de estas diferencias, los patrones de metilación también podrían verse alterados, pues muchos factores asociados a los complejos SWI/SNF se han relacionado con actividades de regulación de la expresión mediante modificación de los mismos. Mediante el ensayo de modificación del DNA por bisulfito sódico estudiamos los patrones de metilación de cinco regiones. Sorprendentemente, el ZBS de w y de dpp, así como las regiones próximas a ellos, se encontraban hipometilados en las hembras z1. El desconocimiento general sobre la metilación en D. melanogaster nos hizo plantear cuales pueden ser las secuencias diana de la metilación en esta especie. Así realizamos un estudio estadístico sobre cuales son las dos bases anteriores y posteriores a las Cs metiladas en nuestras secuencias. Se obtuvieron diferencias en las frecuencias de cada posición y se estableció como secuencia más frecuente para ser metilada ApDp5mCpDpD. Aún así, es una secuencia muy degenerada, y se hacen necesarios estudios más profundos y amplios para confirmarla. / The present PhD thesis studies several zeste1 mutants and this mutation effect over the transcriptional regulation of some genes. The first part is based on the study of the males of M115 and RM115 strains, mainly characterized by the z1 mutation and the insertion of a FB-NOF element in the third intron of the CG32795 gene, known to form a head-to-tail gene pair with white. To study the FB-NOF insertion effect on a z1 background firstly we need to characterize the CG32795, its mRNA sequence and the codified protein predicted structure and function. Several splicing variants were found, not only for its 5' end but also for its 3'end, similar but different from the ones been predicted previously. Once obtained this information we proceeded to study the effects of the FB-NOF insertion over the w and the CG32795 gene expression. The results lead us to a deeper study of the w expression in different body segments, taking into account the zeste-white interaction tissue specificity. Thus, we proved that the w gene expression is not affected by the FB-NOF insertion downstream of its 3' end. The differences observed in the w expression levels were due to the duplication of the w Zeste Binding Site in a z1 background. On the contrary, the FB-NOF insertion not only does modify CG32795 expression, but also mediates the reordenation produced in the RM115 strain being responsible of the w original copy lost and leaving alone the w copy duplicated in the M115 strain.The second part studies the z1 female mutants. We analyzed the expression of two genes possessing a ZBS (w and dpp). The results obtained showed an expression reduction for both genes in the z1 females. Moreover, through the study of the CG32795 gene expression we showed that the zeste-white interaction effect is local, not having any effect on CG32795 expression even though its 5' end is located at only 700bp from the w gene 3' end. Knowing that the Zeste aggregates in the ZBS recruit the chromatin remodelling complex BRM, facilitating thus transcription, we thought that the expression reduction might occur due to chromatin structure alterations. A micrococal nuclease assay was performed and some modifications in nucleosome positioning were found in w and dpp ZBS, being the positioning in the z1 females stricter. Were the BRM complex responsible of those differences, the DNA methylation patterns might also be changed, since many SWI/SNF associated factors have been related to expression regulation duties through DNA methylation patterns modification. A bisulphite modification assay was performed, studying the DNA methylation pattern of five different regions. Surprisingly, w and dpp ZBS and the regions surrounding them were hypomethylated in the z1 females. The little information available about DNA methylation in D. melanogaster encouraged us to study the DNA methylation sites in this specie. Our sequences were statistically analyzed including the two bases before and after the methylated cytosine. The results showed differences in the each nucleotide frequency in each position. The preferently methylated "consensus sequence": ApDp5mCpDpD. However, it is a much degenerated sequence and deeper and broader studies are required to confirm it.

The objective of this research was to design, assemble, and demonstrate the initial performance of a fluidized bed chemical vapor deposition (FB-CVD) system capable of producing thin, uniform zirconium nitride (ZrN) coatings (1 to 10 micrometers thick) on uranium-molybdenum (UMo) particulate fuel. Plate-type fuel with U-xMo (x = 3 to 10 wt.%) particle fuel dispersed in an aluminum matrix is under development at Idaho National Laboratory (INL) for the Reduced Enrichment for Research and Test Reactors (RERTR) program. Initial irradiation tests performed at INL in the Advanced Test Reactor (ATR) indicate an interaction layer forms between the fuel microspheres and the matrix at relatively high power levels. These power levels induce higher temperatures which enables uranium diffusion into the aluminum during irradiation, eventually causing fuel plate failure. The objective of this work was to create a process to mitigate the fuel/matrix interaction by forming a thin barrier coating on the surface of the U-xMo microspheres before incorporation into the dispersion fuel plate matrix. One of the main challenges in performance of the FB-CVD system was the effective fluidization of a powder whose physical characteristics (size, density) are continuously changing. To address this, two types of fluidized bed reaction vessels were designed and improved over the course of this research: a spouted fluidized bed and an inverted fluidized bed. Both reaction vessels utilized tetrakis(dimethylamino)zirconium (TDMAZ) and ammonia gas as precursors at atmospheric pressure. Tungsten wires and zirconia-silica (ZrO2-SiO2) microspheres were used as the substrates for the coating experiments. The substrate temperature and precursor gas flow were manipulated as the process variables. The FB-CVD system was successful in forming zirconium based coatings on surrogate microspheres with elevated levels of chemical impurities. At atmospheric pressure, coatings of thicknesses ranging from 0.5 micrometers to 1.5 micrometers were produced between temperatures of 250 degrees C and 350 degrees C. The deposited coatings were characterized using scanning electron microscopy, energy dispersive spectroscopy and wavelength dispersive spectroscopy.

O sistema complemento parece ter surgido com o aparecimento de C3 e fator B (FB), os únicos componentes encontrados nos organismos mais primitivos, o que sugere que a via alternativa seja a mais antiga. Fragmentos de cDNA codificantes para FB (Lox-FB) e C3 (Lox-C3) foram sintetizados a partir do RNA total isolado da glândula de veneno de Loxosceles laeta e amplificados por técnicas de RACE-PCR. Lox-FB apresenta uma organização de domínios clássica, composta por domínios CCP, vWFA e de serino protease. Os aminoácidos envolvidos na ligação ao C3b são conservados, no entanto, a tríade catalítica clássica não foi encontrada. Lox-C3 apresenta uma configuração de domínios similar a do C3 humano, contendo dois sítios putativos de processamento: um entre as cadeias α e γ, e outro entre as cadeias α e β, indicando que Lox-C3 seja composto por três cadeias. As análises filogenéticas indicaram que Lox-C3 e Lox-FB são mais próximos evolutivamente aos componentes equivalentes da aranha Hasarius adansoni, com valores de identidade de 53% e 43%, respectivamente. / The complement system seems to have arisen with the appearance of C3 and factor B (FB), the only components found in the most primitive organisms, suggesting that the alternative pathway is the oldest. cDNA fragments coding for the complement factor B (Lox-FB) and C3 (Lox-C3) were synthesized from total RNA Loxosceles laeta venom gland and amplified using RACE-PCR techniques. Lox-FB has a classical domain organization in mosaic, composed by CCPs, vWFA and serine protease domains. The amino acids involved in binding to C3b are conserved, however, the classical calatytic triad was not found. Lox-C3 presents a similar configuration of domains to C3 human and has two putative processing sites: the first one is located between α and γ chains and other between α and β chains, indicating that Lox-C3 is composed by three chains. The phylogenetic analyses indicated that Lox-C3 and Lox-FB are evolutionary closer to the equivalent components of Hasarius adansoni, with identity values of 53% and 43%, respectively.

O sistema complemento parece ter surgido com o aparecimento de C3 e fator B (FB), os únicos componentes encontrados nos organismos mais primitivos, o que sugere que a via alternativa seja a mais antiga. Fragmentos de cDNA codificantes para FB (Lox-FB) e C3 (Lox-C3) foram sintetizados a partir do RNA total isolado da glândula de veneno de Loxosceles laeta e amplificados por técnicas de RACE-PCR. Lox-FB apresenta uma organização de domínios clássica, composta por domínios CCP, vWFA e de serino protease. Os aminoácidos envolvidos na ligação ao C3b são conservados, no entanto, a tríade catalítica clássica não foi encontrada. Lox-C3 apresenta uma configuração de domínios similar a do C3 humano, contendo dois sítios putativos de processamento: um entre as cadeias α e γ, e outro entre as cadeias α e β, indicando que Lox-C3 seja composto por três cadeias. As análises filogenéticas indicaram que Lox-C3 e Lox-FB são mais próximos evolutivamente aos componentes equivalentes da aranha Hasarius adansoni, com valores de identidade de 53% e 43%, respectivamente. / The complement system seems to have arisen with the appearance of C3 and factor B (FB), the only components found in the most primitive organisms, suggesting that the alternative pathway is the oldest. cDNA fragments coding for the complement factor B (Lox-FB) and C3 (Lox-C3) were synthesized from total RNA Loxosceles laeta venom gland and amplified using RACE-PCR techniques. Lox-FB has a classical domain organization in mosaic, composed by CCPs, vWFA and serine protease domains. The amino acids involved in binding to C3b are conserved, however, the classical calatytic triad was not found. Lox-C3 presents a similar configuration of domains to C3 human and has two putative processing sites: the first one is located between α and γ chains and other between α and β chains, indicating that Lox-C3 is composed by three chains. The phylogenetic analyses indicated that Lox-C3 and Lox-FB are evolutionary closer to the equivalent components of Hasarius adansoni, with identity values of 53% and 43%, respectively.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

El presente trabajo trata sobre dos temas coyunturalmente relacionados: el fenotipo de las cepas M115 y RM115 de Drosophila melanogaster y el elemento transponible FB-NOF.La cepa M115 de D. melanogaster presenta el fenotipo zeste1 ampliado a los machos. Este fenotipo se caracteriza por los ojos de color amarillo claro y, aunque normalmente se presenta en hembras pues es necesario el apareamiento de dos copias del gen white para producirse, puede observarse en machos portadores de duplicaciones de white. Es la conocida interacción zeste-white. M115 es una cepa inestable y se pueden encontrar individuos revertientes de forma regular. Un macho revertiente dio lugar a la cepa RM115. Este par de cepas son fenotípicamente parecidas a las descritas por la Dra. Rasmuson-Lestander, w+UZ y w+UR.Siguiendo los pasos de Rasmuson-Lestander en la caracterización de w+UZ y w+UR, identificamos la inserción de un elemento transponible FB-NOF en nuestras cepas M115 y RM115, pocas kilobases a 3' del gen white. Aunque el punto de inserción es exactamente el mismo que en las cepas de Rasmuson-Lestander, nuestros experimentos de Southern Blot demuestran que se trata de inserciones distintas, lo cual plantea si esa región podría ser un hot spot de integración para FB-NOF. Al llevar el análisis un paso más adelante, encontramos que las cepas de ojos amarillos, M115 y w+UZ, eran portadoras de una duplicación del gen white, que explica el fenotipo ampliado a los machos. La reversión en RM115, por su parte, consiste en la deleción de una de las dos copias del gen. Así pues, debemos descartar las hipótesis de Rasmuson-Lestander que apuntaban a la interferencia de FB-NOF en la interacción zeste-white.El elemento transponible FB-NOF es uno de los menos conocidos de D. melanogaster. Dio origen al grupo de transposones denominados foldback por su estructura modular repetitiva y se desconocen los detalles de su biología. En este trabajo proponemos una forma más sencilla de estructurar el elemento que facilita su estudio, sobretodo a partir de secuencias genómicas. Además, analizamos su distribución en el genoma de D. melanogaster, la relación entre las secuencias que lo integran y la expresión de su secuencia codificante. Los resultados de dicho análisis revelan que FB-NOF funciona como un solo transposón con un sesgo en sus preferencias de inserción y que se mantiene activo en la actualidad. / The subject of the present thesis focuses on two related issues: the phenotype of the mutant strains M115 and RM115 from Drosophila melanogaster and the transposable element FB-NOF.The D. melanogaster strain M115 presents an extended zeste1 phenotype to both males and females. This phenotype confers a pale yellow color in the eyes and, however it is usually seen only in females since it is necessary to have paired copies of the white gene, this phenotype can also be observed in males carrying a white duplication. This is known as the zeste-white interaction. M115 is an unstable strain and one can find revertant flies regularly. A revertant male established the so-called RM115 strain. These two strains are phenotipically similar to those described by Dr. Rasmuson-Lestander, w+UZ and w+UR.Following Rasmuson-Lestander's steps in the characterization of w+UZ and w+UR, we identified the insertion of an FB-NOF transposable element in M115 and RM115, just few kilobases downstream the white gene's coding region. Despite the insertion point is exactly the same in both sets of strains, ours and Rasmuson-Lestander's, our Southern blot analysis demonstrate that the two insertions are different. This suggests we could have found a hot spot for the FB-NOF integration. Taking our analysis a step further, we found that the yellow-eyed strains, M115 and w+UZ, carried a white gene duplication, which is the main cause for the male extended zeste1 phenotype. Reversion of the phenotype consists in deletion of one of the two copies of the gene. Therefore, we must discard Rasmuson-Lestander's hypothesis pointing to an FB-NOF interference in the zeste-white interaction as the main cause of the extended phenotype.The FB-NOF transposable element is one of the less known in D. melanogaster. It established the group called foldback transposons due to its modular and repetitive structure, and its biology still remains unknown. In this thesis, we propose a simplest way to arrange the element's sequence to make its study from genomic sequences easier. Moreover, we analyze its distribution in the D. melanogaster's genome, the relationship between the sequences that compose it and the expression of the coding regions within. The results from this analysis reveal that FB-NOF behaves as a single transposon, with a bias in its insertion preferences and it is still active at the present time.

Este trabalho apresenta a modelagem matemática e a simulação de potenciais de ação de unidades motoras de músculos de vertebrados visando a posterior simulação do eletromiograma. Para conseguir isso, inicialmente se fez uma compilação de dados existentes para a distribuição das fibras musculares (FBs) nas unidades motoras (MUs) de vários músculos, e as modelagens matemáticas descritos na literatura para o potencial de ação de uma FB (SFAP) e de uma MU (MUAP). Com base nos dados fisiológicos, primeiro se localizou as FBs em um músculo, por meio de uma aproximação de que as FBs estão rodeadas de outras seis no músculo. Para conseguir isto se construiram hexágonos concêntricos por MU, e posteriormente se localizou as FBs nas MUs, cobrindo uma faixa entre 75 e 2000 FBs, o que corresponde a músculos distais de mamíferos. Depois se fez uma aproximação para a distribuição de 170000 FBs nas 272 MUs da cabeça medial do músculo gastrocnêmio (MG) do gato, conseguindo numa primeira simulação localizar cerca de 70% das FBs para cada MU. Com esta localização das FBs no músculo baseados nos dados da literatura se aproximaram os retardos axonais por uma distribuição gaussiana, com média de 2 ms (gato) ou 10 ms (homem) e com desvio padrão de menos de 0,5 ms, desprezando o atraso axonal nas ramificações axonais, que foi estimado no máximo 29 vezes menor. Para a geração do SFAP trabalhou-se com dois modelos, um analítico, o qual resulta em simulações numéricas demoradas, e, outro numérico baseado na convolução da corrente com uma função peso. Para o modelo numérico dobrou-se imaginariamente o comprimento das FBs, para levar em conta o erro computacional de fim de fibra. O modelo numérico resultou em um tempo de simulação 30 vezes menor que o analítico. Adicionalmente, para simular a captação externa (i.e. na pele), fez-se uma aproximação para a função que modela os eletrodos de superfície de secção circular localizados a uma distância maior que 1,79 mm das FBs, mostrando um espectro similar ao reportado na literatura. Finalmente, os MUAPs obtidos resultavam com formas de onda e espectros similares ao descrito na literatura. Além disto, em certos casos, obtiveram-se MUAPs com indentações, seja localizando as junções neuromusculares em bandas da ordem de 1 mm de espessura, seja quando o tempo de atraso axonal foi considerado junto com a velocidade de condução da FB em função da raiz quadrada do diâmetro da FB. Foram feitas simulações para os MG e bíceps braquial do homem. Neste último caso, foram obtidos MUAPs similares aos captados para pessoas saludáveis, e foi observada a freqüência de disparos dos potenciais de ação do motoneurônio no espectro do MUAP. Quanto às formas dos agrupamentos das FBs em uma MU, não se obtiveram diferenças significativas para as FBs posicionadas homogênea e aleatoriamente, exceto uma ligeira variação nas amplitudes. No entanto, ocurreu uma mudança na faixa espectral, quando as FBs estavam concentradas. / This work presents the mathematical model and simulation of motor unit action potentials of vertebrate muscles aiming at after simulation of the electromyogram. To obtain this, initially, it was made a compilation of several data about the distribution of muscle fibers (FBs) in motor units (MUs) of many muscles, and the mathematical models of the action potential of a single FB (SFAP) and MU (MUAP), reported in previous works. On the basis of this physiological data, first, the FB was located in a muscle, using an approximation in which the FBs are encircled with other six FBs in the muscle. To reach this, concentric hexagons were constructed to build the surface of the MU, and later the FBs were situated in the MU, covering a range between 75 and 2000 FBs, corresponding to mammals extremity muscles. Later, a new approximation were was madein order to distribute the 170000 FBs in the 272 MUs of the medial head of muscle medialis gastrocnemius (MG) of the cat, reaching, in a first simulation, the localization of almost 70% of the FBs at each MU. With the FBs lalready allocated in the muscle, and based in data of previous works, their axonal delay were approximated by a gaussian distribution, with mean of 2 ms (cat) or 10 ms (man) and standard deviation of less than 0,5 ms, discarding the axonal delay in the axonal branching, that were estimated to affectup to 29 times less. To SFAP generation, two models were used, the first analytical, resulting in delayed numerical simulations, and the other based on convolution of the second derivate of the current with a weight function, where the length of the FBs was imaginarily duplicated, in order to consider the end fiber effect. Using this, a simulation time 30 times lesser than the analytical one was obtained. Additionally, so as to simulate the external recording (i.e. in the skin), it was made an approximation to the function that models the circular shape surface electrodes located at distances greater than 1,79 mm of the FBs, showing a similar spectrum reported. Finally, the waves and spectrum of the simulated MUAPs resulted similar to the ones reported in the literature. Beyond this, in certain cases, MUAPs were simulated with some tuned, either locating the neuromuscular junctions with thickness bands of 1 mm, or, when the axonal delay and the FB muscle fiber conduction velocity were considered as a function of the square root fiber diameter. This was simulated for MUAPs of MG and biceps brachii muscles of human beings, in the last case it has reached the waveforms and tuned found in heath subjects, and it was visualized the mean frequency of firing rate at the spectrum. In order to know how much affects grouping for the FBs to waves a MU, they were not found significant differences with FBs located homogeneously and randomly, except a little variation in the amplitude of the MUAP. However, they presented a change in the spectral bandwidth when the FBs are more concentrated.

Este trabalho apresenta a modelagem matemática e a simulação de potenciais de ação de unidades motoras de músculos de vertebrados visando a posterior simulação do eletromiograma. Para conseguir isso, inicialmente se fez uma compilação de dados existentes para a distribuição das fibras musculares (FBs) nas unidades motoras (MUs) de vários músculos, e as modelagens matemáticas descritos na literatura para o potencial de ação de uma FB (SFAP) e de uma MU (MUAP). Com base nos dados fisiológicos, primeiro se localizou as FBs em um músculo, por meio de uma aproximação de que as FBs estão rodeadas de outras seis no músculo. Para conseguir isto se construiram hexágonos concêntricos por MU, e posteriormente se localizou as FBs nas MUs, cobrindo uma faixa entre 75 e 2000 FBs, o que corresponde a músculos distais de mamíferos. Depois se fez uma aproximação para a distribuição de 170000 FBs nas 272 MUs da cabeça medial do músculo gastrocnêmio (MG) do gato, conseguindo numa primeira simulação localizar cerca de 70% das FBs para cada MU. Com esta localização das FBs no músculo baseados nos dados da literatura se aproximaram os retardos axonais por uma distribuição gaussiana, com média de 2 ms (gato) ou 10 ms (homem) e com desvio padrão de menos de 0,5 ms, desprezando o atraso axonal nas ramificações axonais, que foi estimado no máximo 29 vezes menor. Para a geração do SFAP trabalhou-se com dois modelos, um analítico, o qual resulta em simulações numéricas demoradas, e, outro numérico baseado na convolução da corrente com uma função peso. Para o modelo numérico dobrou-se imaginariamente o comprimento das FBs, para levar em conta o erro computacional de fim de fibra. O modelo numérico resultou em um tempo de simulação 30 vezes menor que o analítico. Adicionalmente, para simular a captação externa (i.e. na pele), fez-se uma aproximação para a função que modela os eletrodos de superfície de secção circular localizados a uma distância maior que 1,79 mm das FBs, mostrando um espectro similar ao reportado na literatura. Finalmente, os MUAPs obtidos resultavam com formas de onda e espectros similares ao descrito na literatura. Além disto, em certos casos, obtiveram-se MUAPs com indentações, seja localizando as junções neuromusculares em bandas da ordem de 1 mm de espessura, seja quando o tempo de atraso axonal foi considerado junto com a velocidade de condução da FB em função da raiz quadrada do diâmetro da FB. Foram feitas simulações para os MG e bíceps braquial do homem. Neste último caso, foram obtidos MUAPs similares aos captados para pessoas saludáveis, e foi observada a freqüência de disparos dos potenciais de ação do motoneurônio no espectro do MUAP. Quanto às formas dos agrupamentos das FBs em uma MU, não se obtiveram diferenças significativas para as FBs posicionadas homogênea e aleatoriamente, exceto uma ligeira variação nas amplitudes. No entanto, ocurreu uma mudança na faixa espectral, quando as FBs estavam concentradas. / This work presents the mathematical model and simulation of motor unit action potentials of vertebrate muscles aiming at after simulation of the electromyogram. To obtain this, initially, it was made a compilation of several data about the distribution of muscle fibers (FBs) in motor units (MUs) of many muscles, and the mathematical models of the action potential of a single FB (SFAP) and MU (MUAP), reported in previous works. On the basis of this physiological data, first, the FB was located in a muscle, using an approximation in which the FBs are encircled with other six FBs in the muscle. To reach this, concentric hexagons were constructed to build the surface of the MU, and later the FBs were situated in the MU, covering a range between 75 and 2000 FBs, corresponding to mammals extremity muscles. Later, a new approximation were was madein order to distribute the 170000 FBs in the 272 MUs of the medial head of muscle medialis gastrocnemius (MG) of the cat, reaching, in a first simulation, the localization of almost 70% of the FBs at each MU. With the FBs lalready allocated in the muscle, and based in data of previous works, their axonal delay were approximated by a gaussian distribution, with mean of 2 ms (cat) or 10 ms (man) and standard deviation of less than 0,5 ms, discarding the axonal delay in the axonal branching, that were estimated to affectup to 29 times less. To SFAP generation, two models were used, the first analytical, resulting in delayed numerical simulations, and the other based on convolution of the second derivate of the current with a weight function, where the length of the FBs was imaginarily duplicated, in order to consider the end fiber effect. Using this, a simulation time 30 times lesser than the analytical one was obtained. Additionally, so as to simulate the external recording (i.e. in the skin), it was made an approximation to the function that models the circular shape surface electrodes located at distances greater than 1,79 mm of the FBs, showing a similar spectrum reported. Finally, the waves and spectrum of the simulated MUAPs resulted similar to the ones reported in the literature. Beyond this, in certain cases, MUAPs were simulated with some tuned, either locating the neuromuscular junctions with thickness bands of 1 mm, or, when the axonal delay and the FB muscle fiber conduction velocity were considered as a function of the square root fiber diameter. This was simulated for MUAPs of MG and biceps brachii muscles of human beings, in the last case it has reached the waveforms and tuned found in heath subjects, and it was visualized the mean frequency of firing rate at the spectrum. In order to know how much affects grouping for the FBs to waves a MU, they were not found significant differences with FBs located homogeneously and randomly, except a little variation in the amplitude of the MUAP. However, they presented a change in the spectral bandwidth when the FBs are more concentrated.

För att på ett enkelt sätt beskriva det trepartsförhållande som existerar mellan barn, föräldrar och socialnämnd kan sägas att föräldrar har det främsta ansvaret för att barns hälso- och sjukvårdsbehov tillgodoses och ytterst är socialnämnden ansvarig. Nuvarande rättsläge kan ur ett tillämpningsperspektiv beskrivas som en trestegsraket. Den inleds efter att en motiverad vårdnadshavare söker socialnämndens samtycke för att den andre vårdnadshavaren motsätter sig barnets vård. I ett första steg ska socialnämndens utredningsarbete bestå av att ta hänsyn till huvudregeln i föräldrabalken som stadgar att vårdnadshavare har gemensam bestämmanderätt. Socialnämnd ska i och med huvudregeln försöka få vårdnadshavarna att enas i frågan. I andra hand sker ett övervägande om undantagsregeln i 6 kap. 13 a § FB ska användas. I det här läget ska nämnden bedöma vilken förälders inställning har godast skäl för sig, i förhållande till barnets bästa. När den vårdnadshavare som samtycker har goda skäl för sin inställning och barnet har ett påtagligt och tydligt behov av vård kan nämnden istället för den motsägande föräldern samtycka. I sista hand om ingen av vårdnadshavarna samtycker till behövlig vård och förutsättningarna i övrigt är uppfyllda, är det tvångslagen LVU som aktualiseras. Efter att nämnden ansökt hos förvaltningsdomstol kan barnet få vård enligt LVU. Om en av föräldrarna är att anse som lämplig vårdnadshavare kan socialnämnden även väcka talan om ändring av vårdnaden enligt 6 kap. 7 § FB. Som flera av remissinstanserna påpekat i förarbetena till 6 kap. 13 a § FB är det i många av fallen så att vårdnadshavarna har mer långtgående och djupa samarbetssvårigheter än att problemet stannar vid oenighet i en fråga rörande barnet. När förhållandena vårdnadshavarna emellan är så dåliga att det inte föreligger verkliga förutsättningar för gemensam vårdnad är åtminstone jag kritisk till vilken egentlig funktion bestämmelsen fyller. Som framgår i framställningen utgör bestämmelsen en risk för negativa effekter på möjligheterna att ensam förordnas vårdnaden i de situationer förutsättningarna är uppfyllda men domstolen väljer att se 6 kap. 13 a § FB som ett verktyg att använda vid gemensam vårdnad. Syftet bakom bestämmelsens införande är delvis att se till att barn ska slippa invänta ett vårdnadsbeslut från en domstol för att få vård och behandling men följderna för det enskilda barnet kan bli mer långgående än ett uteblivet vård- och behandlingstillfälle. Dagens utformning och tolkning av bestämmelsen medför i min mening en stor brist. Bristen grundar sig i inskränkningen av samförståndskravet. Lagstiftaren accepterar att vårdnadshavare är oeniga i vissa frågor och det sänder ut signaler om att huvudregeln om vårdnadshavares enighet nu försvagats. Regeln måste utvärderas närmare för att det ska kunna avgöras om den fyller den funktion som regeln är avsedd att ha.

La performance mécanique des pièces de structures automobiles fabriquées à partir de tôles d'acier à très haute résistance (THR) est souvent réduite à cause des bords découpés. Ce phénomène a été étudié pour deux nuances d'aciers ferrite-bainite (FB600) et ferrite-martensite (DP600), ce dernier présente un écrouissage et un gradient de dureté entre les phases plus élevés que ceux de la nuance FB600. Les micromécanismes d'endommagement de ces matériaux de base ont été caractérisés en utilisant les techniques de tomographie in situ et MEB in situ. Pour l'acier DP600, la germination de cavités a eu lieu sur les inclusions et aux interfaces ferrite-martensite. De plus, des cavités sous forme d'aiguille ont été observées dans la zone centrale correspondant à la ligne de ségrégation. Les mêmes mécanismes de germination ont été observés dans le cas de l'acier FB en plus de la germination aux interfaces des carbures qui a eu lieu à des déformations élevées. L'analyse d'image a montré que l'acier DP présente une densité initiale de cavités et une densité de cavités germées plus élevées que celles de l'acier FB qui semblait plus tolérant à l'endommagement. Des bords poinçonnés et usinés des nuances DP et FB ont été caractérisés par laminographie in situ lors d'un chargement mécanique. Pour les bords poinçonnés, ces observations ont permis de constater que la zone rompue est rugueuse et qu'un micro-endommagement sous forme d'aiguille initié sur la surface et dans le volume suit les lignes d'écoulement. Lors du chargement mécanique, les cavités sous forme d'aiguilles croissent à partir de la zone rompue et coalescent avec la zone cisaillée. En revanche, pour les bords usinés, l'endommagement s'initie loin de la surface de bords (~800 microns). Une analyse des données 3D a été réalisée pour quantifier l'état initial de l'endommagement et son évolution. L'acier FB600 a été plus résistant aux bords découpés que l'acier DP600. Des simulations 3D par éléments finis ont été menées pour étudier les champs mécaniques potentiellement affectés par le profil du bord découpé et du pré-écrouissage. Cette analyse a permis de conclure que seuls ces paramètres ne modifient pas localement les champs mécaniques. Finalement, des simulations axisymétriques par éléments finis de l'essai d'expansion de trou ont été réalisées pour différentes épaisseurs de tôle en utilisant les critères d'endommagement identifiés sur les résultats expérimentaux de la tomographie in situ. / The mechanical properties of automotive structures made of advanced high strength steels (AHSS) is often seen reduced by the presence of cut edges. Here this phenomenon is investigated for ferrite-bainite steel (FB600) and martensite ferrite steel (DP600), the latter having higher work hardening and phase hardness gradient than FB600.Damage micromechanisms for these two base materials were assessed using in situ synchrotron tomography, in situ SEM and SEM on cross sections. It was revealed for the DP600 steel that damage nucleated from particles and ferrite-martensite interfaces. In addition, needle shaped voids, that are consistent with the presence of segregation lines, were seen. For the FB steel, the same observations hold true except that the decohesion on interfaces sets in at higher strains. Quantitative image analysis also showed that the initial number of voids and the number of nucleating voids was higher for DP steel than for FB steel which was also seen to be more damage tolerant.Punched and machined edges made of DP600 and FB600 steel were mechanically loaded during in situ laminography testing. It was found that the fracture zone of the punched edge was rough and that needle-shape voids at the surface and in the bulk followed material flow lines. During mechanical in situ testing the needle voids grew from the fracture zone surface and coalesced with the sheared zone. In contrast, for the machined edge the damage started away from the edge (~ 800 microns) where substantial necking has occurred. Three-dimensional image analysis was performed to quantify the initial damage and its evolution. The FB600 was more resistant to cut edges than the DP600 steel.3D elasto-plastic FE calculations were carried out to investigate mechanical fields, potentially affected by the edge profile and pre-hardening profile. These parameters were not found to substantially modify the mechanical fields. Finally, axisymmetric 2D simulations for hole expansion were carried out for different sheet thicknesses using a post-treated damage evaluation calibrated on in-situ tomography data.