Diss. Linköping : Univ., 2003.

Thesis (Ph. D.)--Illinois State University, 1974. / Title from title page screen, viewed Oct. 28, 2004. Dissertation Committee: H.E. Brockman, A.G. Richardson (co-chairs), A.E. Liberta, H.W. Huizinga, D. McCracken, F. Schwalm. Includes bibliographical references (leaves 119-140) and abstract. Also available in print.

Thesis (Ph. D.)--Lund University, 1995. / Published dissertation.

Thesis (Ph. D.)--Lund University, 1995. / Published dissertation.

La Tachycardie Ventriculaire Polymorphe Catécholaminergique (TVPC) est une pathologie rythmique héréditaire rare et sévère, responsable de mort subite chez le sujet jeune. Les mutations des gènes RYR2 et CASQ2 sont associées respectivement à une transmission autosomique dominante et récessive de la maladie. Le canal calcique RyR2 et la protéine chélatrice du calcium Casq2 sont situés dans le réticulum sarcoplasmique (RS) où ils participent au complexe de relâchement calcique (CRC), essentiel à l'homéostasie calcique cardiaque. L'analyse de RYR2 et CASQ2 chez 214 probands ayant présenté une TVPC nous a permis d'identifier respectivement des mutations chez 75 et 11 probands. Deux cas de mosaïques germinales et somatiques ont été identifiés dans le gène RYR2. Deux mutations d'épissage du gène CASQ2 ont été validées à l'aide de minigènes. Chez 97 patients négatifs pour RYR2 et CASQ2, nous avons décidé de rechercher des mutations de trois protéines du CRC (la triadine, la junctine et FKBP12.6) en séquençant les gènes correspondants. Nous n'avons retrouvé aucune mutation de la junctine, ni de FKBP12.6. En revanche, nous avons identifié trois mutations de la triadine: une micro-délétion et une mutation non-sens entraînant un codon stop prématuré, ainsi qu'une variation faux-sens, dont la caractérisation à l'aide de modèle animal et cellulaire a montré qu'elle entraînait une dégradation massive de la protéine. Les mutations du gène TRDN seraient associées à une absence de triadine entraînant une dysfonction du CRC, à l'origine des arythmies observées. En conclusion, nos résultats confirment que RYR2 est le gène majeur impliqué dans la TVPC, CASQ2 étant rarement impliqué; et nous rapportons, pour la première fois, des mutations du gène TRDN en pathologie humaine, associée à une forme autosomique rare de TVPC. / Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a rare and severe inherited arrhythmogenic disorder, responsible for sudden death in young patients. It is a genetically heterogenous pathology with an autosomal dominant form associated with mutations of the RYR2 gene, and a recessive form associated with mutations of the CASQ2 gene. The ryanodine receptor RyR2 is a Ca2+ channel, and the calsequestrin Casq2 is the major calcium storage protein, located in the sarcoplasmic reticulum of the cardiomyocytes. They belong to the calcium release complex (CRC) that plays a central role in excitation-contraction coupling. In this work, we report the identification of RYR2 and CASQ2 mutations in 75 and 11 CPVT probands, respectively. We identified two cases of germline and somatic mosaicism in RYR2. Two splicing mutations of CASQ2 have been validated using a splicing minigene assay. We searched for mutations among 97 CPVT probands, negative for RYR2 and CASQ2, in three candidate genes: TRDN, ASPH and FKBP1B, encoding three proteins of the CRC. We did not identify any mutation of ASPH and FKBP1B genes. However, we found three mutations in the TRDN gene, encoding the cardiac triadin: a microdeletion, a nonsense mutation, both leading to a premature stop codon, and a missense mutation. We demonstrated that the missense mutation induces a drastic reduction of the protein in cellular and animal models. All the three mutations would thus be associated with the absence of triadin, leading to dysfunction of the CRC, and arythmias. In conclusion, our results confirm that RYR2 is the major gene implicated in CPVT, and CASQ2 rarely implicated. Moreover, we report mutations of the TRDN gene for the first time in pathology, as a third gene associated with a rare autosomal recessive form of CPVT.

Made available in DSpace on 2014-12-17T14:16:20Z (GMT). No. of bitstreams: 1 GiocondaDRL.pdf: 1031402 bytes, checksum: 0016e69e527bddffea93909fa2748a03 (MD5) Previous issue date: 2007-08-29 / Hereditary Hemochromatosis (HH) is a genetic disease caused by high iron absorption and deposition in several organs. This accumulation results in clinical disturbances such as cirrhosis, arthritis, cardiopathies, diabetes, sexual disorders and skin darkening. The H63D and C282Y mutations are well defined in the hemochromatosis etiology. The aim of this paper was that of identifying the H63D and C282Y genetical mutations in the hemochromatosis gene and the frequency assessment of these mutations in the HFE protein gene in patients with hyperferritin which are sent to the DNA Center laboratory in Natal, state of Rio Grande do Norte. This paper also evaluates the HH H63D and C282Y gene mutations genotype correlation with the serum ferritin concentration, glucose, alanine aminotransferasis, aspartato aminotransferasis, gama glutamil transferasis and with the clinical complications and also the interrelation with life habits including alcoholism and iron overload. The biochemical dosages and molecule analyses are done respectively by the enzymatic method and PCR with enzymatic restriction. Out of the 183 patients investigated, 51,4% showed no mutation and 48,6% showed some type of mutation: 5,0% were C282Y heterozygous mutation; 1,1%, C282Y homozygous mutation; 31%, H63D heterozygous mutation; 8,7%, H63D homozygous mutation; and 3,3%, heterozygous for the mutation in both genes. As to gender, we observed a greater percentage of cases with molecular alteration in men in relation to women in the two evaluated mutations. The individuals with negative results showed clinical and lab signs which indicate hemochromatosis that other genes could be involved in the iron metabolism. Due to the high prevalence of hemochromatosis and taking into account that hemochromatosis is considered a public health matter, its gravity being preventable and the loss treatment toxicity, the early genetic diagnosis is indicated, especially in patients with high ferritin, and this way it avoids serious clinical manifestations and increases patients' life expectation. Our findings show the importance of doing such genetic studies in individuals suspected of hereditary hemochromatosis due to the high incidence of such a hereditary disease in our region / A hemocromatose heredit?ria (HH) ? uma doen?a gen?tica causada pela absor??o e deposi??o elevada de ferro em v?rios ?rg?os. Este ac?mulo resulta em complica??es cl?nicas como cirrose, artrite, cardiopatias, diabetes, desordens sexuais e escurecimento da pele. As muta??es H63D e C282Y est?o bem definidas na etiologia da hemocromatose. O objetivo deste trabalho foi a identifica??o das muta??es gen?ticas H63D e C282Y no gene da Hemocromatose e avalia??o da freq??ncia dessas muta??es no gene da prote?na HFE em pacientes com hiperferritinemia que s?o encaminhados ao laborat?rio DNA Center Natal / RN. Al?m disso, avaliar a correla??o dos gen?tipos das muta??es H63D e C282Y do gene da HH com a concentra??o s?rica da ferritina, glicose, alanina aminotransferase, aspartato minotransferase, gt e com as complica??es cl?nicas e ainda a interrela??o com os h?bitos de vida incluindo o etilismo e dieta com sobrecarga de ferro. As dosagens bioqu?micas e an?lises moleculares foram realizadas respectivamente atrav?s do m?todo enzim?tico e PCR com restri??o enzim?tica. Dos 183 pacientes investigados 51,4% apresentaram aus?ncia de muta??o e 48,6% com algum tipo de muta??o: 5,0% C282Y heterozigoto mutado; 1,1% C282Y homozigoto mutado; 31% H63D heterozigoto mutado; 8,7% H63D homozigoto mutado; e 3,3% heterozigoto para a muta??o em ambos os genes. Com rela??o ao sexo, observou-se o maior percentual de casos com altera??o molecular em homens em rela??o a mulheres nas duas muta??es avaliadas. Os indiv?duos com resultados negativos apresentaram sinais cl?nicos e laboratoriais indicativos de hemocromatose sugerindo que outros genes poder?o estar envolvidos no metabolismo do ferro. Devido ? alta preval?ncia da hemocromatose, e tendo em vista que a hemocromatose ? considerada um problema de sa?de p?blica, sua gravidade ser preven?vel e a baixa toxicidade do tratamento, o diagn?stico gen?tico precoce torna-se indicado, principalmente nos pacientes com ferritina elevada, e com isso evitar manifesta??es cl?nicas graves e aumentar a expectativa de vida dos pacientes com esta doen?a. Nossos achados mostram a import?ncia da realiza??o de estudos gen?ticos em indiv?duos com suspeita de hemocromatose heredit?ria em virtude de elevada incid?ncia dessa doen?a de cunho heredit?rio em nossa regi?o

No desenvolvimento de software, a qualidade do produto está diretamente relacionada à qualidade do processo de desenvolvimento. Diante disso, atividades de Verificação, Validação & Teste (VV&T) realizadas por meio de métodos, técnicas e ferramentas são de extrema necessidade para o aumento da produtividade, qualidade e diminuição de custos no desenvolvimento de software. Do mesmo modo, técnicas e critérios contribuem para a produtividade das atividades de teste. Um ponto crucial para o teste de software é sua automatização, tornando as atividades mais confiáveis e diminuindo significativamente os custos de desenvolvimento. Na automatização dos testes, os oráculos são essenciais, representando um mecanismo (programa, processo ou dados) que indica se a saída obtida para um caso de teste está correta. Este trabalho de mestrado utiliza a ideia de mutação para criar implementações alternativas de oráculos de teste e, assim, avaliar a sua qualidade. O teste de mutação se refere à criação de versões do sistema em desenvolvimento com pequenas alterações sintáticas de código. A mutação possui alta eficácia na detecção de defeitos e é bastante flexível na sua aplicação, podendo ser utilizada em diversos tipos de artefatos. Adicionalmente, este trabalho propõe operadores de mutação específicos para oráculos, implementa uma ferramenta de apoio à utilização desses operadores para oráculos e também descreve um estudo empírico dos operadores, destacando benefícios e desafios associados ao seu uso. / In software development, product quality is directly related to the quality of the development process. Therefore, activities of Verification, Validation & Testing (VV&T) performed by methods, techniques and tools are urgently required to increase productivity, quality and cost reduction in software development. Similarly, testing technique and criteria contribute to the productivity of test activities. A crucial point for the software testing automation is making the most reliable activities and significantly reducing development costs. Regarding software testing automation, test oracles are essential, representing an mechanism (program, process or data) to indicate whether the actual output for a given test case is correct. This masters thesis aims to explore concepts of mutation testing to create alternative implementations of the oracle procedure and thus assess their quality. Mutation testing refers to the creation of system development versions with minor syntactic code changes. It has high efficiency on defects detecting and it is very flexible in its application and it is being used in various types of artifacts. This work also proposes specific mutation operators for oracles, implements an useful support tool for using these oracle mutation operators and conducts an empirical study of operators, highlighting benefits and challenges associated with their use.

Um dos pontos fundamentais na atividade de teste de software é o projeto de casos de teste. Diversos critérios de adequação têm sido propostos com o objetivo de fornecer meios que permitam que a avaliação e elaboração de casos de teste sejam feitas de maneira sistemática e fundamentadas teoricamente. Infelizmente, a maioria dos critérios de adequação de casos de teste definidos tem seu uso restrito ao teste de unidade. Para fases posteriores da atividade de teste, em particular para o teste de integração, nota-se a ausência de critérios de adequação, principalmente porque os critérios propostos definem requisitos de teste que se restringem aos limites de uma única unidade, não exercitando de maneira efetiva as interações entre as unidades, que devem ser alvo principal no teste de integração. Com exceção de alguns poucos trabalhos que procuram estender critérios estruturais para o nível interprocedimental, tem-se utilizado nessa fase de teste, quase que exclusivamente, critérios funcionais. Dada essa ausência de critérios e salientando ainda o caráter complementar entre as diferentes técnicas de teste, esta tese apresenta um critério de teste interprocedimental baseado em defeitos chamado de Mutação de Interface. Esse critério busca exercitar as interações entre as unidades através da seleção de casos de teste que distingam mutantes criados pela introdução de defeitos típicos e que, de acordo com um modo definido, caracterizamos erros de integração. Definiu-se um conjunto de operadores de Mutação de Interface que concentram sua aplicação em pontos do programa relacionados com as interações entre as unidades, como, por exemplo, chamadas de subprogramas e seus parâmetros. Dados o alto custo de aplicação, inerente de critérios baseados em mutação, e pelas próprias características do conjunto de operadores de Mutação de Interface, torna-se necessário definirem-se abordagens para reduzir esse custo. Assim, foram estabelecidas maneiras de se parametrizar a aplicação dos operadores de mutação, definindo-se critérios de Mutação de Interface alternativos, estendendo-se abordagem sutilizadas no teste de mutação convencional como mutação restrita. A aplicação de um critério de teste está fortemente condicionada à sua automatização. A definição de um critério de teste sem que pelo menos se apontem soluções para sua automatização tem pouca utilidade prática. Por isso, especificou-se e implementou-se a ferramenta PROTEUM/IM para apoiar a aplicação do critério Mutação de Interface. Essa ferramenta torna-se essencial neste trabalho à medida que permite que estudos empíricos possam ser realizados, avaliando o critério proposto. Dois estudos de caso são apresentados. Esses estudos aplicam o critério Mutação de interface em programas reais e buscam avaliar seu custo e sua eficácia em revelar erros. Estes estudos aplicam ainda critérios alternativos, mostrando que a Mutação de interface é bastante efetiva em revelar erros o de ter custo de aplicação bastante reduzido, quando aplicada de maneira incremental, utilizando-se as parametrizações que os operadores de mutação oferecem. / The project of test cases is one of the most important topics in the software testing activity. Several criteria have been proposed aiming at allowing the evaluation and selection of test cases in a systematic and theoretically well founded way. Unfortunately, the use of most of these criteria is restricted to the unit testing phase. For other testing phases, in particular for integration testing. there is a lack of such criteria, mainly because the existing criteria define test requirements only in the scope of a single unit. They arc not able to effectively exercise the interactions between units, what should be the focus of integration testing. Excepting some few works that extend structural criteria to the interprocedural level, only functional testing has been used at integration testing phase. Given this lack of criteria and the complementary characteristics of different testing techniques, this thesis presents an interprocedural fault based criterion named Interface Mutation. This criterion exercises the interactions between units through the selection of test cases that distinguish mutants created by introducing typical faults that characterize integration errors. A set of Interface Mutation operators was defined. The focus of these operators are the points of the program related to the unit interactions, for instance, subprogram calls and their parameters. Given the high cost associated to mutation testing in general and particularly to the Interface Mutation operators, it is necessary to define some approaches to reduce its application cost. Thus, some parameterizations were defined to the mutation operators, allowing to establish alternative Interface Mutation criteria, extending approaches already used in conventional mutation testing, as random mutation and constrained mutation. The application of any testing criterion strongly depends on its automatization. The definition of a criterion, without pointing out ways to its implementation has little practical utility. So, a tool named PROTEUM/IM was specified and implemented to support the application of Interface Mutation. This tool is an essential point in the present work because it allows the conduction of empirical studies aiming at evaluating the proposed criterion. Two case studies arc presented. In these studies the criterion Interface Mutation is applied to real programs and the cost of its application as well as its errors revealing effectiveness are evaluated. Alternative criteria are also used. Showing that Interface Mutation is very effective to reveal errors and can be applied with a reduced cost if used in an incremental way, taking advantage of the parameterization characteristics provided by the Interface Mutation operators set.

Les déficits en facteurs de la coagulation sont des pathologies hémorragiques congénitales rares. Le développement récent des techniques de biologie moléculaire ont permis l'indentification de nombreuses anomalies génétiques dans les gènes codant les facteurs de coagulation. Cependant, la détermination de l'impact clinique réel de ces nouveaux variants est souvent un défi pour le biologiste moléculaire.La première partie de ce travail a consisté à l'étude par analyse chromosomique sur puce à ADN de grands réarrangements génomiques identifiés chez des patients hémophiles A ou B ayant parfois des phénotypes cliniques atypiques. Nous avons montré que certains gènes voisins des gènes F8 et F9 étaient associés, lorsqu'ils sont concernés par ces réarrangements de grande taille, à des déficits mentaux (SOX3) ou des pathologies cardiovasculaires (BRCC3).La seconde partie de cette étude a été centrée sur l'étude des variants de signification indéterminée localisés au niveau des sites d'épissage. Nous avons démontré par l'utilisation conjointe d'algorithmes informatiques et de tests in vitro de type minigène la pathogénicité de 21 variants identifiés chez des hémophiles A.La dernière partie de ce travail a consisté en la description de nouveaux mécanismes moléculaires responsables de pathologies hémorragiques. Nous avons identifié une délétion intronique chez 6,1% des hémophiles A mineurs de notre cohorte. Nous avons montré que cette anomalie était probablement responsable d'une dérégulation de l'hnRNP C ce qui entrainait l'insertion d'une séquence AluY dans les transcrits du gène F8. Enfin, nous avons décrit les mécanismes moléculaires responsables de la présence de concentrations très élevées de thrombomoduline soluble dans les plasmas de patients porteur de la mutation du gène THBD c.1611C>A. Ces travaux ont permis de détecter et de mieux appréhender certains mécanismes moléculaires responsables de pathologies rares hémorragiques / Inherited coagulation disorders are caused by a large number of genetic abnormalities. However, the determination of the clinical impact for some of these genetic variations is challenging for the molecular biologist.In a first part, we characterized large genomic rearrangements in haemophilia patients using cytogentic microarray analysis. In a first study, we delineated six F9 complete deletions in order to investigate genotype/phenotype correlations. We identified SOX3 as a candidate gene for intellectual disability (ID) found Haemophilia B patients. In a second study, we described five complex Xq28 rearrangements in Haemophilia A (HA) patients. We showed that several F8 neighbouring genes are involved in these rearrangements and some of these genes are involved in other pathologies such as ID, physical abnormalities and cardiovascular disease. Such investigations would be particularly useful for genetic counseling in female carriers to assess the risk of transmitting haemophilia associated with other diseases.In a second, we developed a minigene assay to characterise putative splice site mutations in F8 and we showed that 21 out of the 26 variations studied are associated with splicing defect.In the third part, we described two original molecular mechanisms leading to coagulation disorders. In a first case, we reported a recurrent deep intronic deletion in mild HA. We gave some evidences that this deletion promoted AluY exonization in F8 transcrits. Due to its high prevalence (6.1%), this deletion must be investigated in all patients with mild HA in whom no mutation has been detected by standard genetic analysis. In the second study we investigated the mechanism responsible for bleeding tendency in patient carrying the THBD c.1611C>A and having high levels of soluble thrombomoduline (TM). We showed that a higher sensitivity of the mutated TM to the proteolysis by metalloproteases and a defect of TM synthesis seemed responsible for TM shedding

Résumé. Les derniers jours de l'Enseignement ?Analyse de la fonction enseignante, cette thèse de philosophie de l'éducation pose la question des conditions pour que l'enseigner ait encore du sens et un avenir. Elle montre l'évolution, la mutation des modalités, comme la déconstruction et le délitement d'une « forme scolaire » menacée car trop pesante, donc aussi la transformation nécessaire de l'enseignement.A partir de l'exemple des ateliers-philo, des déplacements de l'enseignant depuis la transmission jusqu'à l'animation et l'accompagnement, et de l'hypothèse que notre époque n'a pas (encore) pris en compte ce que le moderne a porté au monde, occulté par les représentations antérieures ou cadres d'interprétation passés, elle s'interroge sur ce qui hante les conceptions de l'enseignement ou la pensée pédagogique pour en tirer quelques conséquences quant au devenir de cette fonction dans ses nouvelles conditions.Pour ce faire, ce travail interroge l'enseignement dans ses définitions comme dans ses transformations récentes, par rapport au nouveau statut de l'enfance ou de l'adolescence et au malaise dans la transmission jusqu'à la crise de la représentation, pour le mettre en perspective par rapport aux conflits mondiaux et à ce qu'ils « disent » de l'évolution de la pensée occidentale, qu'il semble nécessaire d'intégrer à la perception critique de l'école. Donc il resitue l'enseignement dans son rapport à la fin de la métaphysique : quels sont les effets de ce changement d'époque sur la condition enseignante ? Et il rapporte les troubles dans l'institution scolaire et universitaire à sa provenance et son histoire : à quelles conditions la ou les fonctions de l'enseignant peuvent-elles perdurer dans le monde d'après les conflits mondiaux et le nazisme, d'après la crise des fondements ?En reconnaissant l'interruption du continuum de la tradition et du progrès continu, donc le fossé, la brisure, les failles et traces toujours déjà laissées qui font la désorientation, peut-on et comment réinventer le « professer » / le profiteor, et sa capacité instituante, au moins déceler les « signes » d'un renouvellement dans une perspective de déconstruction de la structure retardataire, secondarisée, de l'enseignement ? L'évolution et la crise enseignante étant symptôme d'un changement plus profond, qu'il nous faut regarder en face, avec une vision qui va du philosophique à sa pensée au-delà, oeuvrer à la mutation en cours exige une autre écriture de la geste enseignante.Peut-être que l'enseignant n'enseigne pas encore… / Summary. The last days of Teaching?Analysis of the role of Teaching, this philosophy thesis asks the question on the necessary conditions in order, for education, to keep a meaning and a future. It describes the evolution, the mutation of methods, as well as the deconstruction and disintegration of an « academic form », threatened because too burdensome, and therefore the necessary transformation of education.Basing itself on the example of the philosophy workshop, on the movements of the teacher from the transmission to the animation and the support, and on the theory that our time has not (yet) taken into account what modernity has brought to the world, overshadowed by the prior representations or past interpretations, it examines what haunts the conceptions of education or pedagogic thought in order to draw consequences as to the future of this function in its new conditions.To this end, this work questions Teaching in its definitions as well as in its recent transformations, with regard to childhood or adolescence new status and the unease of transmission until the representation crisis, to put it in perspective with worldwide conflicts and to what they « say » of the evolution of the occidental way of thinking, which seems necessary to integrate to the critical perception of school. Thus it replaces teaching in its relation to the end of metaphysics: what are the effects of this change of era on the teaching condition ? And it relates the disorders in the scholar and university institution to its origin and history : under what conditions can the teachers functions persist in the world after nazism and world conflicts, after the foundation crisis ?By recognizing the interruption between the continuum of tradition and the evergoing progress, thus the gap, the cracks, marks always already left which cause the desorientation, can we and by what means, reinvent the « teaching »/the profiteor, and its institutionalizing capacity, at least detect the « signs » of a renewal with, in prospect, the deconstruction of the outdated, second-rated, structure of teaching? The evolution and teaching crisis are symptoms of a deeper change that we must face, with a vision which includes philosophy but also its thought beyond, act for the ongoing mutation will require another writing of the teaching gesture.Maybe the teacher isn't teaching yet…