QC 351 A7 no. 39 / Optical systems known as "null correctors" are often required to test certain aspheric optical surfaces. This report classifies these systems on the basis of their first -order geometry and analyzes the merits of each type. The behavior of optical aberrations, especially spherical aberration, in these systems is examined in the context of computer optimization techniques, particular attention being given to some design problems unique to null correcting systems. Orthonormal concepts are applied to the problem of reducing spherical aberration in null correctors. It is shown that exceedingly simple merit functions may be constructed to streamline the optimization process. These merit functions are composed of simple linear sums of the angular spherical aberration coefficients B1, B3, B5, and B7. Thus, minimizing the following sums will improve nearly diffraction - limited systems: ( - 13 B1 + 1 B3 - g' B5 - B7) , ( 4.131 - B3 - B5) , ( - 2B1 - B3) , and ( - B1) /1-5- 3/7 3 or ( 120 B3 + 960 B5 + 840 B7 ) , ( 840 B5 + 2520 B7) , and ( 840 B7) Non -diffraction - limited systems may be optimized by minimizing the sums ( 6 B3 + 5 B5 + 5 B7) , ( p B5 + 3 B7) , and ( 1 0 B7) To demonstrate the effectiveness of the techniques discussed, the process of designing a specific null correcting system is followed in detail.

Abstract not available. / text

Cystic Fibrosis (CF) is an autosomal recessive disease that arises from mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. The deletion of phenylalanine-508 (ΔF508-CFTR) is the most prevalent CF mutation and results in a misfolded protein that fails to exit the endoplasmic reticulum (ER). Previous studies demonstrated that mutation of a di-arginine based ER retention motif (R553AR555) in the first nucleotide binding domain (NBD1) rescues the trafficking defect of ΔF508-CFTR. We hypothesized that if the R553AR555 motif mediates retention of the ΔF508-CFTR protein, peptides that mimic this motif should antagonize mistrafficking mediated by aberrant exposure of the endogenous R553AR555 motif. We generated a peptide bearing the R553AR555 motif (CF-RXR) and conjugated it to the cell penetrating peptide Tat (CPP-CF-RXR) to facilitate intracellular delivery and investigated its efficacy in rescuing the mistrafficking and function of ΔF508-CFTR. Using a variety of biochemical and functional assays we demonstrate that the CPP-CF-RXR peptide is effective at increasing surface expression of ΔF508-CFTR in baby hamster kidney (BHK) and human embryonic kidney (HEK) cell lines. Furthermore, the increased surface expression is accompanied by an increase in its functional expression as a chloride channel. Using Ussing chamber assays, we demonstrate that the CPP-CF-RXR peptide improved ΔF508-CFTR channel function in respiratory epithelial tissues obtained from CF patients. Additionally, we investigated the effects of small molecules on mediating biosynthetic rescue of a ΔF508-CFTR construct bearing the additional mutations R553K and R555K (ΔFRK-CFTR) to inactivate the R553AR555 motif. Interestingly, mutation of the R553AR555 motif exerts an additive effect with correctors VRT-325 and Corrector 4a. Taken together, our data suggests that abnormal accessibility of the RXR motif present in NBD1 is a key determinant of the mistrafficking of the major CF causing mutant.

Neste trabalho estudamos a continuidade da dinâmica assintótica relativamente a perturbações e, em particular, exploramos a obtenção de melhorias para as taxas de convergência de atratores globais através da introdução de fatores de correção, inspirados pelos resultados da teoria de homogeneização e nos trabalhos de (BABIN; VISHIK, 1992) e (CARVALHO; CHOLEWA, 2011), e através da introdução de mecanismos que melhoram a transferência da taxa de convergência de semigrupos para a taxa de convergência de atratores, inspirados pelos trabalhos (SANTAMARÍA, 2013) e (BABIN; VISHIK, 1992; CARVALHO; CHOLEWA, 2011). A proposta inicial está centrada na obtenção de melhores taxas de convergência de atratores globais através da obtenção de equiatração e da melhoria da taxa de convergência dos semigrupos. Para isto, buscamos melhorar a taxa de convergência do resolvente dos operadores setoriais envolvidos, por meio de uma perturbação singular do resolvente limite que ainda gere uma família de operadores setoriais com resolventes que aproximam o resolvente do problema limite e aproximam melhor os resolventes das perturbações iniciais. Feito isto, obtemos uma melhora imediata de convergência dos semigrupos lineares, depois dos não lineares (através da fórmula da variação das constantes). Motivados pelos resultados de (SANTAMARÍA, 2013), que oferecem uma menor perda na transferência das taxas de convergência dos semigrupos para as taxas de convergência dos atratores, buscamos melhor compreender a propriedade Lipschitz Shadowing, que é responsável direta pela obtenção da taxa de convergência dos atratores diretamente da taxa de convergência dos semigrupos. Isto nos levou a descobrir que podemos obter as propriedade Lipschitz Shadowing e estabilidade estrutural para perturbações Lipschitz de semigrupos Morse-Smale. / Here we compare the continuity of the asymptotic dynamics with respect to perturbations and, in particular, we explored to obtain improvement of rates of convergence of the global attractor through the introduction of correction factors, inspired by the results of homogenization theory and work of (BABIN; VISHIK, 1992) and (CARVALHO; CHOLEWA, 2011), and the introduction of mechanisms that improve the transference of the convergence rate of semigroups to the convergence rate of attractors, inspired by the work of (SANTAMARÍA, 2013) and (BABIN; VISHIK, 1992; CARVALHO; CHOLEWA, 2011). The initial proposal is focused on achieving best rates of convergence of the global attractors by obtaining equi-atraction and improving the convergence rate of semigroups. For this, we seek to improve the rate of convergence of the resolvents of sectorial operators, through a singular perturbation of the resolvent associated with the limit problem and generate a new family of sectorial operators whose resolvents both approximate the resolvent of the limit problem as they were closer to the resolvents the initial perturbation. Having done this, we obtain an immediate improvement of convergence of linear semigroups, after the non-linear (using the variation of constants formula). Motivated by the results of (SANTAMARÍA, 2013), which offer an improvement in obtaining convergence rates, we seek to study property better Lipschitz Shadowing, which is basically responsible for obtaining the distance of the attractors directly from the convergence rate of the semigroups. This has led us to discover that we can both preserve the Lipschitz Shadowing property under Lipschitz perturbations of Morse-Smale semigroups, and The geometric stability of the attractors.

The degradation of astronomical images caused by atmospheric turbulence will be much more severe in the next generation of terrestrial telescopes and its compensation will require deformable mirrors with up to tens-of-thousands of actuators. Current designs for these correctors consist of scaling up the proven technologies of flexible optical plates deformed under the out-of-plane action of linear actuators. This approach will lead to an exponential growth of cost with the number of actuators, and in very complex mechanisms. This thesis proposes a new concept of optical correction which is modular, robust, lightweight and low-cost and is based on the bimorph in-plane actuation. The adaptive mirror consists of segmented identical hexagonal bimorph mirrors allowing to indefinitely increase the degree of correction while maintaining the first mechanical resonance at the level of a single segment and showing an increase in price only proportional to the number of segments. Each bimorph segment can be mass-produced by simply screen-printing an array of thin piezoelectric patches onto a silicon wafer resulting in very compact and lightweight modules and at a price essentially independent from the number of actuators. The controlled deformation of a screen-printed bimorph mirror was experimentally achieved with meaningful optical shapes and appropriate amplitudes; its capability for compensating turbulence was evaluated numerically. The generation of continuous surfaces by an assembly of these mirrors was numerically simulated and a demonstrator of concept consisting of 3 segments was constructed.

Cystic Fibrosis (CF) is an autosomal recessive disease that arises from mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. The deletion of phenylalanine-508 (ΔF508-CFTR) is the most prevalent CF mutation and results in a misfolded protein that fails to exit the endoplasmic reticulum (ER). Previous studies demonstrated that mutation of a di-arginine based ER retention motif (R553AR555) in the first nucleotide binding domain (NBD1) rescues the trafficking defect of ΔF508-CFTR. We hypothesized that if the R553AR555 motif mediates retention of the ΔF508-CFTR protein, peptides that mimic this motif should antagonize mistrafficking mediated by aberrant exposure of the endogenous R553AR555 motif. We generated a peptide bearing the R553AR555 motif (CF-RXR) and conjugated it to the cell penetrating peptide Tat (CPP-CF-RXR) to facilitate intracellular delivery and investigated its efficacy in rescuing the mistrafficking and function of ΔF508-CFTR. Using a variety of biochemical and functional assays we demonstrate that the CPP-CF-RXR peptide is effective at increasing surface expression of ΔF508-CFTR in baby hamster kidney (BHK) and human embryonic kidney (HEK) cell lines. Furthermore, the increased surface expression is accompanied by an increase in its functional expression as a chloride channel. Using Ussing chamber assays, we demonstrate that the CPP-CF-RXR peptide improved ΔF508-CFTR channel function in respiratory epithelial tissues obtained from CF patients. Additionally, we investigated the effects of small molecules on mediating biosynthetic rescue of a ΔF508-CFTR construct bearing the additional mutations R553K and R555K (ΔFRK-CFTR) to inactivate the R553AR555 motif. Interestingly, mutation of the R553AR555 motif exerts an additive effect with correctors VRT-325 and Corrector 4a. Taken together, our data suggests that abnormal accessibility of the RXR motif present in NBD1 is a key determinant of the mistrafficking of the major CF causing mutant.

Neste trabalho estudamos a continuidade da dinâmica assintótica relativamente a perturbações e, em particular, exploramos a obtenção de melhorias para as taxas de convergência de atratores globais através da introdução de fatores de correção, inspirados pelos resultados da teoria de homogeneização e nos trabalhos de (BABIN; VISHIK, 1992) e (CARVALHO; CHOLEWA, 2011), e através da introdução de mecanismos que melhoram a transferência da taxa de convergência de semigrupos para a taxa de convergência de atratores, inspirados pelos trabalhos (SANTAMARÍA, 2013) e (BABIN; VISHIK, 1992; CARVALHO; CHOLEWA, 2011). A proposta inicial está centrada na obtenção de melhores taxas de convergência de atratores globais através da obtenção de equiatração e da melhoria da taxa de convergência dos semigrupos. Para isto, buscamos melhorar a taxa de convergência do resolvente dos operadores setoriais envolvidos, por meio de uma perturbação singular do resolvente limite que ainda gere uma família de operadores setoriais com resolventes que aproximam o resolvente do problema limite e aproximam melhor os resolventes das perturbações iniciais. Feito isto, obtemos uma melhora imediata de convergência dos semigrupos lineares, depois dos não lineares (através da fórmula da variação das constantes). Motivados pelos resultados de (SANTAMARÍA, 2013), que oferecem uma menor perda na transferência das taxas de convergência dos semigrupos para as taxas de convergência dos atratores, buscamos melhor compreender a propriedade Lipschitz Shadowing, que é responsável direta pela obtenção da taxa de convergência dos atratores diretamente da taxa de convergência dos semigrupos. Isto nos levou a descobrir que podemos obter as propriedade Lipschitz Shadowing e estabilidade estrutural para perturbações Lipschitz de semigrupos Morse-Smale. / Here we compare the continuity of the asymptotic dynamics with respect to perturbations and, in particular, we explored to obtain improvement of rates of convergence of the global attractor through the introduction of correction factors, inspired by the results of homogenization theory and work of (BABIN; VISHIK, 1992) and (CARVALHO; CHOLEWA, 2011), and the introduction of mechanisms that improve the transference of the convergence rate of semigroups to the convergence rate of attractors, inspired by the work of (SANTAMARÍA, 2013) and (BABIN; VISHIK, 1992; CARVALHO; CHOLEWA, 2011). The initial proposal is focused on achieving best rates of convergence of the global attractors by obtaining equi-atraction and improving the convergence rate of semigroups. For this, we seek to improve the rate of convergence of the resolvents of sectorial operators, through a singular perturbation of the resolvent associated with the limit problem and generate a new family of sectorial operators whose resolvents both approximate the resolvent of the limit problem as they were closer to the resolvents the initial perturbation. Having done this, we obtain an immediate improvement of convergence of linear semigroups, after the non-linear (using the variation of constants formula). Motivated by the results of (SANTAMARÍA, 2013), which offer an improvement in obtaining convergence rates, we seek to study property better Lipschitz Shadowing, which is basically responsible for obtaining the distance of the attractors directly from the convergence rate of the semigroups. This has led us to discover that we can both preserve the Lipschitz Shadowing property under Lipschitz perturbations of Morse-Smale semigroups, and The geometric stability of the attractors.

La mucoviscidose est la maladie génétique récessive la plus fréquente dans les populations caucasiennes. Elle est dûe à des mutations dans le gène CFTR qui code pour une protéine qui a une fonction canal chlorure et qui est exprimée au pôle apical des épithéliums sécrétoires. La mutation la plus fréquente F508del altère le repliement de la protéine, induisant sa dégradation précoce par le protéasome et son absence à la membrane plasmique. La recherche fondamentale se focalise sur la protéine mutée et cherche à découvrir des molécules capables d’adresser celle-ci au pôle apical des cellules épithéliales où elle pourra remplir sa fonction de canal chlorure. Au sein du laboratoire, nous nous intéressons aux interactions de la protéine CFTR/ F508del-CFTR avec d’autres protéines. Nous avons déjà montré l’existence d’une interaction non voulue entre F508del-CFTR et un filament intermédiaire, la cytokératine 8. En combinant une approche in-silico et du criblage haut débit, nous avons identifié des molécules capables d’interrompre ces interactions. Des tests fonctionnels sur des lignées cellulaires ainsi que sur des modèles murins ont montré une restauration de la fonction canal chlorure après traitements avec ces molécules. Au cours de ma thèse, je me suis intéressée à l’une d’entre elles qui est la molécule « c407 ». L’objectif de ma thèse était de comprendre les mécanismes d’action de cette molécule. J’ai également évalué l’efficacité des traitements actuels dans le contexte des allèles complexes de F508del-CFTR. Dans une seconde partie de ma thèse, j’ai étudié l’effet d’une cytokine (TNFα) sur la protéine mutée F508del-CFTR. De façon inattendue, j’ai observé que le TNFα, à des concentrations physiologiques, corrige le défaut de routage de la protéine F508del-CFTR. Cette observation pourrait expliquer une fonction résiduelle de la F508del-CFTR chez certains patients atteints de mucoviscidose. En conclusion, mes travaux ont permis de préciser les mécanismes d’action d’un correcteur et de découvrir un effet inattendu d’une cytokine pro-inflammatoire. Ces travaux permettent de relier la correction d’un défaut de routage au processus inflammatoire ouvrant ainsi un nouveau champ d’investigation. / Cystic fibrosis is due to the loss of epithelial chloride transport caused by mutations in the CFTR gene, the most frequent mutation being F508del. One of the strategies developed to find new treatment for Cystic fibrosis (CF) is to discover compounds that correct the trafficking of F508del-CFTR to the plasma membrane. Using hypothesis-driven approach and combining modeling of NBD1, molecular docking and functional assays, we identified 4 compounds that correct F508del-CFTR function in cells (including human primary bronchial cells in culture) and F508del mice. New correctors probably act by interrupting the interaction between F508del-CFTR with keratin 8 (Odolczyk et al EMBO Mol Med 2013). During my PhD, I focused on one of those molecules, the "c407" molecule. The aim of my thesis was to investigate the mechanisms of action of this molecule. I have also evaluated the effectiveness of current treatments in the context of complex alleles F508del-CFTR. In the second part of my thesis, I studied the effect of a cytokine (TNFα) on the protein F508del-CFTR. Unexpectedly, I observed that the TNFα at physiological concentrations, corrects the trafficking of F508del-CFTR protein. This observation could explain a residual function of F508del-CFTR in some CF patients. In conclusion, my thesis helped to clarify the mechanisms of action of new correctors of F508del-CFTR.

La mucoviscidose est la maladie génétique récessive la plus fréquente dans les populations caucasiennes. Elle est dûe à des mutations dans le gène CFTR qui code pour une protéine qui a une fonction canal chlorure et qui est exprimée au pôle apical des épithéliums sécrétoires. La mutation la plus fréquente F508del altère le repliement de la protéine, induisant sa dégradation précoce par le protéasome et son absence à la membrane plasmique. La recherche fondamentale se focalise sur la protéine mutée et cherche à découvrir des molécules capables d’adresser celle-ci au pôle apical des cellules épithéliales où elle pourra remplir sa fonction de canal chlorure. Au sein du laboratoire, nous nous intéressons aux interactions de la protéine CFTR/ F508del-CFTR avec d’autres protéines. Nous avons déjà montré l’existence d’une interaction non voulue entre F508del-CFTR et un filament intermédiaire, la cytokératine 8. En combinant une approche in-silico et du criblage haut débit, nous avons identifié des molécules capables d’interrompre ces interactions. Des tests fonctionnels sur des lignées cellulaires ainsi que sur des modèles murins ont montré une restauration de la fonction canal chlorure après traitements avec ces molécules. Au cours de ma thèse, je me suis intéressée à l’une d’entre elles qui est la molécule « c407 ». L’objectif de ma thèse était de comprendre les mécanismes d’action de cette molécule. J’ai également évalué l’efficacité des traitements actuels dans le contexte des allèles complexes de F508del-CFTR. Dans une seconde partie de ma thèse, j’ai étudié l’effet d’une cytokine (TNFα) sur la protéine mutée F508del-CFTR. De façon inattendue, j’ai observé que le TNFα, à des concentrations physiologiques, corrige le défaut de routage de la protéine F508del-CFTR. Cette observation pourrait expliquer une fonction résiduelle de la F508del-CFTR chez certains patients atteints de mucoviscidose. En conclusion, mes travaux ont permis de préciser les mécanismes d’action d’un correcteur et de découvrir un effet inattendu d’une cytokine pro-inflammatoire. Ces travaux permettent de relier la correction d’un défaut de routage au processus inflammatoire ouvrant ainsi un nouveau champ d’investigation. / Cystic fibrosis is due to the loss of epithelial chloride transport caused by mutations in the CFTR gene, the most frequent mutation being F508del. One of the strategies developed to find new treatment for Cystic fibrosis (CF) is to discover compounds that correct the trafficking of F508del-CFTR to the plasma membrane. Using hypothesis-driven approach and combining modeling of NBD1, molecular docking and functional assays, we identified 4 compounds that correct F508del-CFTR function in cells (including human primary bronchial cells in culture) and F508del mice. New correctors probably act by interrupting the interaction between F508del-CFTR with keratin 8 (Odolczyk et al EMBO Mol Med 2013). During my PhD, I focused on one of those molecules, the "c407" molecule. The aim of my thesis was to investigate the mechanisms of action of this molecule. I have also evaluated the effectiveness of current treatments in the context of complex alleles F508del-CFTR. In the second part of my thesis, I studied the effect of a cytokine (TNFα) on the protein F508del-CFTR. Unexpectedly, I observed that the TNFα at physiological concentrations, corrects the trafficking of F508del-CFTR protein. This observation could explain a residual function of F508del-CFTR in some CF patients. In conclusion, my thesis helped to clarify the mechanisms of action of new correctors of F508del-CFTR.

La mucoviscidose est la maladie génétique récessive la plus fréquente dans les populations caucasiennes. Elle est dûe à des mutations dans le gène CFTR qui code pour une protéine qui a une fonction canal chlorure et qui est exprimée au pôle apical des épithéliums sécrétoires. La mutation la plus fréquente F508del altère le repliement de la protéine, induisant sa dégradation précoce par le protéasome et son absence à la membrane plasmique. La recherche fondamentale se focalise sur la protéine mutée et cherche à découvrir des molécules capables d’adresser celle-ci au pôle apical des cellules épithéliales où elle pourra remplir sa fonction de canal chlorure. Au sein du laboratoire, nous nous intéressons aux interactions de la protéine CFTR/ F508del-CFTR avec d’autres protéines. Nous avons déjà montré l’existence d’une interaction non voulue entre F508del-CFTR et un filament intermédiaire, la cytokératine 8. En combinant une approche in-silico et du criblage haut débit, nous avons identifié des molécules capables d’interrompre ces interactions. Des tests fonctionnels sur des lignées cellulaires ainsi que sur des modèles murins ont montré une restauration de la fonction canal chlorure après traitements avec ces molécules. Au cours de ma thèse, je me suis intéressée à l’une d’entre elles qui est la molécule « c407 ». L’objectif de ma thèse était de comprendre les mécanismes d’action de cette molécule. J’ai également évalué l’efficacité des traitements actuels dans le contexte des allèles complexes de F508del-CFTR. Dans une seconde partie de ma thèse, j’ai étudié l’effet d’une cytokine (TNFα) sur la protéine mutée F508del-CFTR. De façon inattendue, j’ai observé que le TNFα, à des concentrations physiologiques, corrige le défaut de routage de la protéine F508del-CFTR. Cette observation pourrait expliquer une fonction résiduelle de la F508del-CFTR chez certains patients atteints de mucoviscidose. En conclusion, mes travaux ont permis de préciser les mécanismes d’action d’un correcteur et de découvrir un effet inattendu d’une cytokine pro-inflammatoire. Ces travaux permettent de relier la correction d’un défaut de routage au processus inflammatoire ouvrant ainsi un nouveau champ d’investigation. / Cystic fibrosis is due to the loss of epithelial chloride transport caused by mutations in the CFTR gene, the most frequent mutation being F508del. One of the strategies developed to find new treatment for Cystic fibrosis (CF) is to discover compounds that correct the trafficking of F508del-CFTR to the plasma membrane. Using hypothesis-driven approach and combining modeling of NBD1, molecular docking and functional assays, we identified 4 compounds that correct F508del-CFTR function in cells (including human primary bronchial cells in culture) and F508del mice. New correctors probably act by interrupting the interaction between F508del-CFTR with keratin 8 (Odolczyk et al EMBO Mol Med 2013). During my PhD, I focused on one of those molecules, the "c407" molecule. The aim of my thesis was to investigate the mechanisms of action of this molecule. I have also evaluated the effectiveness of current treatments in the context of complex alleles F508del-CFTR. In the second part of my thesis, I studied the effect of a cytokine (TNFα) on the protein F508del-CFTR. Unexpectedly, I observed that the TNFα at physiological concentrations, corrects the trafficking of F508del-CFTR protein. This observation could explain a residual function of F508del-CFTR in some CF patients. In conclusion, my thesis helped to clarify the mechanisms of action of new correctors of F508del-CFTR.