Les hydrocarbures aromatiques polycycliques (HAP) sont des composés ubiquitaires issus de la combustion incomplète de matières organiques. Ils sont à l'origine de pollutions de l'environnement, surtout liées à l'exploitation des produits pétroliers, car ce sont des composés toxiques pour les êtres vivants et pour l'homme en particulier. De nombreuses bactéries capables de dégrader les HAP ont été isolées et étudiées, mais celles qui les dégradent in situ sont mal connues, car moins de 5% des bactéries du sol sont cultivables en laboratoire. Le premier objectif de cette étude était d'identifier les bactéries qui dégradent les HAP dans le sol par des méthodes moléculaires indépendantes de la culture. A cette fin, une stratégie de marquage isotopique in situ a été mise en œuvre qui repose sur l'utilisation du phénanthrène, un HAP à trois cycles, dans lequel l'isotope naturel du carbone a été remplacé par le 13C. Cette molécule a été introduite comme traceur dans des microcosmes contenant du sol provenant d'un bassin de rétention des eaux de ruissellement d'autoroute. Les bactéries ayant incorporé le 13C ont ensuite été identifiées par séquençage des gènes d'ARNr 16S amplifiés à partir de l'ADN marqué extrait du sol. Les résultats montrent que des Betaprotéobactéries peu étudiées à ce jour, appartenant aux genres Acidovorax, Rhodoferax, Hydrogenophaga et Thiobacillus, ainsi que des Rhodocyclaceae, étaient les principaux acteurs de la dégradation du phénanthrène. La prépondérance des Betaprotéobactéries a été établie par des mesures de PCR quantitative. Une analyse dynamique de la diversité bactérienne a montré que celle-ci changeait en fonction de la biodisponibilité du phénanthrène. En outre, la diversité d'arène-dioxygénases impliquées dans la dégradation des HAP a été explorée sur le plan phylogénétique et fonctionnel. Nous avons ainsi détecté des séquences nouvelles, pour la plupart apparentées à des dioxygénases de Sphingomonadales et de Burkholderiales. Grâce à la construction et l'expression d'enzymes hybrides, il a été possible, pour la première fois, d'associer une activité catalytique d'oxydation des HAP à des séquences partielles de gènes, amplifiées à partir de l'ADN du sol. Les résultats obtenus et les outils mis au point dans cette étude pourront servir à développer des méthodes de diagnostic et de suivi de biodégradation de polluants, par exemple dans le cadre d'opérations de bioremédiation de sites pollués par les HAP. / Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous compounds produced by incomplete combustion of organic matter. They are a source of environmental pollution, especially associated to oil product exploitation, and represent a threat for living organisms including human beings because of their toxicity. Many bacteria capable of degrading PAHs have been isolated and studied. However, since less than 5% of soil bacteria can be cultivated in the laboratory, bacterial species able to degrade PAHs in situ have been poorly studied. The first goal of this study was to identify bacteria that degrade PAHs in soil using culture-independent molecular methods. To this end, a strategy known a stable isotope probing has been implemented based on the use of phenanthrene, a three rings PAH, in which the natural isotope of carbon was replaced by 13C. This molecule has been introduced as a tracer in microcosms containing soil from a constructed wetlands collecting contaminated water from highway runoff. Bacteria having incorporated the 13C were then identified by 16S rRNA gene sequence analysis after PCR amplification from labeled genomic DNA extracted from soil. The results show that so far little studied Betaproteobacteria, belonging to the genera Acidovorax, Rhodoferax, Hydrogenophaga and Thiobacillus, as well as Rhodocyclaceae, were the key players in phenanthrene degradation. Predominance of Betaprotéobactéries was established thanks to quantitative PCR measurements. A dynamic analysis of bacterial diversity also showed that the community structure of degraders depended on phenanthrene bioavailability. In addition, the phylogenetic diversity of ring-hydroxylating dioxygenases, enzymes involved in the first step of PAH degradation, has been explored. We detected new sequences, mostly related to dioxygenases from Sphingomonadales and Burkholderiales. For the first time, we were able to associate a catalytic activity for oxidation of PAHs to partial gene sequences amplified from soil DNA, by constructing hybrid enzymes and assaying their activity The results obtained and the tools implemented in this study may be used to develop methods for the diagnostic and monitoring of pollutant biodegradation in processes such as bioremediation of PAHs contaminated sites.

Proper eye development depends on expression and mutual regulation of many genes. Anterior segment dysgenesis (ASD) are a highly heterogeneous group of diseases exhibiting all types of Mendelian inheritance, which manifest as combination of congenital abnormalities of the cornea, iris, anterior chamber angle or lens. Screening of genes associated with ASD does not often lead to the identification of the underlying genetic cause implying that there are still novel variants or genes to be discovered. Molecular genetic analysis in 12 probands with ASD using Sanger and whole-exome sequencing were performed. Functional analysis by Exon trapping assay was provided in variants predicted to effect pre-mRNA splicing. Four PAX6 mutations evaluated as pathogenic or likely pathogenic in a heterozygous state were found in four probands c.183C˃G; p.(Tyr61*), c.1032+1G>A, c.1183+1G>T and c.622C>T; p.(Arg208Trp). One proband was found to be a compound heterozygote for c.244A>G; p.(Met82Val) and c.541delG; p.(Glu181Lysfs*26) mutations in FOXE3. In 7 probands, no potentially pathogenic variants were identified. Exon trapping assay confirmed that mutations c.1032+1G>A and c.1183+1G>T have an effect on pre-mRNA splicing of the PAX6 gene. Detailed molecular-genetic analysis in patients with ASD may contribute to...

Pathologische Sorgen sind ungenau definiert. Für die Behandlungsplanung bleiben wichtige Fragen offen: Welche Merkmale sind für die Unterscheidung zwischen behandlungsbedürftigen und nicht behandlungsbedürftigen Sorgen relevant? Welche Art von Sorgen muss wie behandelt werden? Und: Welche Art von Sorgen gilt es eher zu akzeptieren? Wir machen praxisnahe Vorschläge dafür, wie Sorgen mittels einer einfachen Heuristik auch vom Patienten selbst als «pathologisch» identifiziert werden können. Im Sinne eines therapeutischen Arbeitsmodells ergeben sich differentielle Bearbeitungsstrategien, je nachdem, ob es sich um wichtige oder weniger wichtige, auf lösbare oder unlösbare Probleme bezogene sowie angemessene oder überzogene Sorgen handelt. Das vorgestellte Arbeitsblatt zu den Sorgen soll vor allem die wahrgenommene Kontrolle des Patienten stärken und die Psychoedukation zur Generalisierten Angststörung erleichtern. / Pathological worries have not yet been clearly defined. As a consequence, practically relevant questions remain open: Which characteristics distinguish worries relevant for treatment from those which are not? What kind of worries has to be treated in which way? And: What kind of worries is rather to be accepted? We propose a simple rationale which helps the therapist and the patient to identify pathological worries. According to this working model, different treatment strategies result depending on whether worries are central or not, whether they relate to a problem which can be solved or not, and whether they seem proportionate or exaggerated. The presented worksheet is meant to strengthen the perceived control of the patient and to help facilitate psychoeducation for generalised anxiety disorder. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

This thesis project was conveyed in collaboration with Block Zero, a strategic Design Studio in Malmö, Sweden, with the purpose to design a product that develops a new type of interaction within the Smart Home. This design project will primarily focus on the research, exploration and definition of possible solutions, and the resulting design and development of the final product, a Smart Home Health Hub. From a global perspective, throughout the following document is covered a description of the design process from initial research to the final prototype. The project is defined in this approach and establishes the mentioned principles to guide the execution of the project through a deductive method, synthesising until the final result.

Indiana University-Purdue University Indianapolis (IUPUI) / Physical and biological phenomena that involve oscillations on multiple time scales attract attention of mathematicians because resulting equations include a small parameter that allows for decomposing a three- or higher-dimensional dynamical system into fast/slow subsystems of lower dimensionality and analyzing them independently using geometric singular perturbation theory and other techniques. However, in most life sciences applications observed dynamics is extremely complex, no small parameter exists and this approach fails. Nevertheless, it is still desirable to gain insight into behavior of these mathematical models using the only viable alternative – ad hoc computational analysis. Current dissertation is devoted to this latter approach. Neural networks in the region of the brain called basal ganglia (BG) are capable of producing rich activity patterns. For example, burst firing, i.e. a train of action potentials followed by a period of quiescence in neurons of the subthalamic nucleus (STN) in BG was shown to be related to involuntary shaking of limbs in Parkinson’s disease called tremor. The origin of tremor remains unknown; however, a few hypotheses of tremor-generation were proposed recently. The first project of this dissertation examines the BG-thalamo-cortical loop hypothesis for tremor generation by building physiologically-relevant mathematical model of tremor-related circuits with negative delayed feedback. The dynamics of the model is explored under variation of connection strength and delay parameters in the feedback loop using computational methods and data analysis techniques. The model is shown to qualitatively reproduce the transition from irregular physiological activity to pathological synchronous dynamics with varying parameters that are affected in Parkinson’s disease. Thus, the proposed model provides an explanation for the basal ganglia-thalamo-cortical loop mechanism of tremor generation. Besides tremor-related bursting activity BG structures in Parkinson’s disease also show increased synchronized activity in the beta-band (10-30Hz) that ultimately causes other parkinsonian symptoms like slowness of movement, rigidity etc. Suppression of excessively synchronous beta-band oscillatory activity is believed to suppress hypokinetic motor symptoms in Parkinson’s disease. Recently, a lot of interest has been devoted to desynchronizing delayed feedback deep brain stimulation (DBS). This type of synchrony control was shown to destabilize synchronized state in networks of simple model oscillators as well as in networks of coupled model neurons. However, the dynamics of the neural activity in Parkinson’s disease exhibits complex intermittent synchronous patterns, far from the idealized synchronized dynamics used to study the delayed feedback stimulation. The second project of this dissertation explores the action of delayed feedback stimulation on partially synchronous oscillatory dynamics, similar to what one observes experimentally in parkinsonian patients. We employ a computational model of the basal ganglia networks which reproduces the fine temporal structure of the synchronous dynamics observed experimentally. Modeling results suggest that delayed feedback DBS in Parkinson’s disease may boost rather than suppresses synchronization and is therefore unlikely to be clinically successful. Single neuron dynamics may also have important physiological meaning. For instance, bistability – coexistence of two stable solutions observed experimentally in many neurons is thought to be involved in some short-term memory tasks. Bistability that occurs at the depolarization block, i.e. a silent depolarized state a neuron enters with excessive excitatory input was proposed to play a role in improving robustness of oscillations in pacemaker-type neurons. The third project of this dissertation studies what parameters control bistability at the depolarization block in the three-dimensional conductance-based neuronal model by comparing the reduced dopaminergic neuron model to the Hodgkin-Huxley model of the squid giant axon. Bifurcation analysis and parameter variations revealed that bistability is mainly characterized by the inactivation of the Na+ current, while the activation characteristics of the Na+ and the delayed rectifier K+ currents do not account for the difference in bistability in the two models.

Proper eye development depends on expression and mutual regulation of many genes. Anterior segment dysgenesis (ASD) are a highly heterogeneous group of diseases exhibiting all types of Mendelian inheritance, which manifest as combination of congenital abnormalities of the cornea, iris, anterior chamber angle or lens. Screening of genes associated with ASD does not often lead to the identification of the underlying genetic cause implying that there are still novel variants or genes to be discovered. Molecular genetic analysis in 12 probands with ASD using Sanger and whole-exome sequencing were performed. Functional analysis by Exon trapping assay was provided in variants predicted to effect pre-mRNA splicing. Four PAX6 mutations evaluated as pathogenic or likely pathogenic in a heterozygous state were found in four probands c.183C˃G; p.(Tyr61*), c.1032+1G>A, c.1183+1G>T and c.622C>T; p.(Arg208Trp). One proband was found to be a compound heterozygote for c.244A>G; p.(Met82Val) and c.541delG; p.(Glu181Lysfs*26) mutations in FOXE3. In 7 probands, no potentially pathogenic variants were identified. Exon trapping assay confirmed that mutations c.1032+1G>A and c.1183+1G>T have an effect on pre-mRNA splicing of the PAX6 gene. Detailed molecular-genetic analysis in patients with ASD may contribute to...

The natural environments of organisms present a multitude of biotic and abiotic challenges that require both short-term ecological and long-term evolutionary responses. Though most environmental response studies have focused on effects at the ecosystem, community and organismal levels, the ultimate controls of these responses are located in the genome of the organism. Soil nematodes are highly responsive to, and display a wide variety of responses to changing environmental conditions, making them ideal models for the study of organismal interactions with their environment. In an attempt to examine responses to environmental stress (desiccation and freezing), genomic level analyses of gene expression during anhydrobiosis of the Antarctic nematode Plectus murrayi was undertaken. An EST library representative of the desiccation induced transcripts was established and the transcripts differentially expressed during desiccation stress were identified. The expressed genome of P. murrayi showed that desiccation survival in nematodes involves differential expression of a suite of genes from diverse functional areas, and constitutive expression of a number of stress related genes. My study also revealed that exposure to slow desiccation and freezing plays an important role in the transcription of stress related genes, improves desiccation and freezing survival of nematodes. Deterioration of traits essential for biological control has been recognized in diverse biological control agents including insect pathogenic nematodes. I studied the genetic mechanisms behind such deterioration using expression profiling. My results showed that trait deterioration of insect pathogenic nematode induces substantial overall changes in the nematode transcriptome and exhibits a general pattern of metabolic shift causing massive changes in metabolic and other processes. Finally, through field observations and molecular laboratory experiments the validity of the growth rate hypothesis in natural populations of Antarctic nematodes was tested. My results indicated that elemental stoichiometry influences evolutionary adaptations in gene expression and genome evolution. My study, in addition to providing immediate insight into the mechanisms by which multicellular animals respond to their environment, is transformative in its potential to inform other fundamental ecological and evolutionary questions, such as the evolution of life-history patterns and the relationship between community structure and ecological function in ecosystems.

Despite the strong internal validity of functional analysis, most behavior analysts continue to use descriptive and indirect methods to conduct functional behavior assessment. However, most descriptive and indirect methods have received little empirical support for their convergent validity with functional analysis. The ABC narrative recording and the Open-Ended Functional Assessment Interview (OEFAI) stand out as descriptive and indirect methods that have received some empirical evidence for their convergent validity, but those studies included some limitations. Thus, this dissertation aimed to evaluate more thoroughly these two approaches. The purpose of the first study was to verify the convergent validity of ABC narrative recordings and of the OEFAI with a functional analysis. One hundred seventy-six behavior analysts analyzed ABC narrative and OEFAI transcripts of challenging behaviors in four children with autism spectrum disorder (ASD). Results show that, for three of four children, the OEFAI conclusions corresponded more with the outcomes of functional analysis than did the ABC narrative recording. Psychoeducators are also involved in the treatment of challenging behaviors in children with ASD. However, we did not find studies that described the performance of psychoeducators in functional behavior assessment. The purpose of the second study was to examine the convergent validity of behavioral functions identified by psychoeducators with the outcomes of functional analysis and to compare them with functions identified by behavior analysts. Fifty-four psychoeducators analyzed the same transcripts used in the first study to identify behavioral functions. Psychoeducators produced more accurate functions when using the OEFAI for three children in four. Behavior analysts were more exact than psychoeducators in four of seven comparisons while the three others remained undifferentiated. Altogether, the results underline the importance of enhancing training in functional behavior assessment for psychoeducators. / Malgré la validité interne élevée de l’analyse fonctionnelle, la majorité des analystes du comportement et des psychoéducateurs continuent d’utiliser les méthodes descriptives et indirectes pour effectuer des évaluations fonctionnelles du comportement. Cependant, la plupart des méthodes descriptives et indirectes ont reçu peu de soutien empirique pour appuyer leur validité convergente. La grille antécédent – comportement – conséquence (ACC) narrative et l’Open-Ended Functional Assessment Interview (OEFAI) se démarquent comme des méthodes descriptives et indirectes dotées de soutien empirique pour appuyer leur validité convergente, mais ces études comportent des faiblesses. Cette thèse vise donc à effectuer une évaluation plus approfondie de ces deux approches. L’objectif de la première étude est d’évaluer la validité convergente de la grille ACC narrative et de l’OEFAI. Nous avons recruté 176 analystes du comportement pour analyser des transcriptions de la grille ACC narrative et de l’OEFAI comportant des données de comportements problématiques de quatre enfants ayant un trouble du spectre de l’autisme (TSA). Les résultats montrent que pour trois enfants analysés sur quatre, les conclusions tirées de l’OEFAI correspondent plus souvent avec celles de l’analyse fonctionnelle que celles de la grille ACC narrative. Les psychoéducateurs interviennent aussi dans la réduction des comportements problématiques chez les enfants ayant un TSA. Toutefois, il est difficile de trouver des études qui décrivent les performances en évaluation fonctionnelle du comportement chez les psychoéducateurs. L’objectif de la seconde étude est d’examiner la validité convergente des fonctions comportementales identifiées par des psychoéducateurs et de les comparer à celles des analystes du comportement. Cinquante-quatre psychoéducateurs ont analysé les mêmes transcriptions de la grille ACC narrative et de l’OEFAI utilisées dans la première étude. Les résultats montrent que les psychoéducateurs sont plus exacts pour identifier la fonction du comportement en utilisant l’OEFAI pour trois des enfants et en utilisant la grille ACC pour un enfant. Les analystes du comportement sont plus exacts que les psychoéducateurs dans quatre comparaisons sur sept tandis que les trois autres sont non différenciées. Les résultats soulignent la nécessité de bonifier les formations en évaluation fonctionnelle du comportement chez les psychoéducateurs.

Obwohl der Wildtyp des Hepatitis-B-Virus (HBV) nicht zytopathogen und die Pathogenese der Hepatitis B generell immunvermittelt ist, können in immunsupprimierten Nierentransplantatempfängern mit chronischer Hepatitis B schwere Leberschäden bis hin zu Leberzirrhose und Leberversagen entstehen. Die Entwicklung von Leberzirrhose in den Nierentransplantierten ist assoziiert mit der Akkumulation und Persistenz von komplexen HBV-Varianten mit Mutationen im Core-Promotor / X-Gen, Deletionen im Core (C)-Gen und teilweise zusätzlichen Deletionen im präS-Bereich. Dies lässt eine Rolle der Varianten in der speziellen Pathogenese bei Immunsupprimierten vermuten. In der vorliegenden Arbeit wurden funktionelle Analysen der komplexen Varianten im Vergleich zu Referenz-Wildtypgenomen und Wildtyp-ähnlichen Genomen der Patienten aus der frühen Infektionsphase durchgeführt, um Hinweise auf den potentiellen Beitrag der Varianten zur Pathogenese zu erlangen. Die Analysen erfolgten durch transiente Transfektion der humanen Hepatomazelllinie HuH7 mit repräsentativen HBV-Gesamtgenomen, die aus 2 Patienten während des Krankheitsverlaufs von einer asymptomatischen Infektion hin zur Leberzirrhose isoliert und kloniert worden waren. Trotz einiger Unterschiede im Detail wiesen die komplexen Varianten einen gemeinsamen, drastisch vom Wildtyp abweichenden Phänotyp auf. Dieser war gekennzeichnet durch eine veränderte Transkription mit reduzierten präC- und Oberflächen-mRNAs und verstärkter Expression der prägenomischen RNA, eine starke Reduktion des häufigsten Spleißprodukts der prägenomischen RNA, SP1, eine extrem reduzierte oder fehlende Expression und/oder Sekretion aller Oberflächenproteine und des HBeAg, ein verändertes intrazelluläres Verteilungsmuster des schwach exprimierten Core-Proteins und teilweise der Oberflächenproteine sowie eine erhöhte Replikation und Anreicherung gegenüber Wildtyp-HBV aufgrund einer verstärkten reversen Transkription der prägenomischen RNA. Dieser Phänotyp basierte zum Teil auf den Mutationen in Core-Promotor und C-Gen, wurde jedoch deutlich durch zusätzliche Mutationen in den übrigen Genomabschnitten beeinflusst. Die vielfältigen Veränderungen der Varianten unterstützen ihren vermuteten Beitrag zur Pathogenese. / Although wild-type hepatitis B virus is not cytopathogenic and the pathogenesis of hepatitis B is generally immune mediated, also immuno-suppressed patients, such as renal transplant recipients, with chronic hepatitis B may develop liver cirrhosis and end-stage liver disease. In renal transplant recipients, the development of liver cirrhosis is associated with the accumulation and persistence of complex HBV variants with mutations in core promoter / X gene, deletions in core (C) gene and sometimes additional deletions in the preS region. This suggests a role of these variants in the special pathogenesis in immuno-suppressed patients. In the present work, the complex variants were functionally analyzed in comparison to reference wild-type genomes and wild-type-like HBV genomes from the early asymptomatic phase of infection. For the analyses, representative cloned full-length HBV genomes isolated from 2 patients before and during liver cirrhosis were transiently transfected into the human hepatoma cell line HuH7. In spite of some variations, the complex variants showed a common phenotype, which was drastically altered compared to wild-type. It was characterized by reduced preC and surface mRNAs and increased expression of pregenomic RNA, by a strong reduction of the major spliced pregenomic RNA, SP1, by a partial or complete defect in expression and/or secretion of surface proteins and HBeAg, by an aberrant intracellular localization of the weakly expressed core protein and in some cases of the surface proteins, and by an enhanced replication and enrichment over wild-type HBV due to an enhanced reverse transcription of variant pregenomic RNA. The phenotypic alterations were often based on the mutations in core promoter and C gene but were considerably influenced by the additional mutations in other genomic regions. The multiple functional changes of the variants support their assumed contribution to pathogenesis.

Cette thèse traite de la résolution théorique et constructive de problèmes inverses pour des équations de diffusion isotropes dans des domaines plan simplement et doublement connexes. A partir de données de Cauchy (potentiel, flux) disponibles sur une partie de la frontière du domaine, il s'agit de retrouver ces quantités sur la partie du bord où l'on ne dispose pas d'information, ainsi qu'à l'intérieur du domaine. L'approche mise au point consiste à considérer les solutions de l'équation de diffusion comme les parties réelles des solutions complexes d'une équation de Beltrami conjuguée. Ces fonctions analytiques généralisées d'un type particulier permettent de définir des classes de Hardy, dans lesquelles le problème inverse est régularisé en étant reformulé comme un problème de meilleure approximation sous contrainte (ou encore problème extrémal borné, d'adéquation aux données). Le caractère bien posé de celui-ci est assuré par des résultats d'existence et de régularité auxquels s'ajoutent des propriétés de densité à la frontière. Une application au calcul de la frontière libre d'un plasma sous confinement magnétique dans le tokamak Tore Supra (CEA-IRFM Cadarache) est proposée. La résolution du problème extrémal à partir d'une base de fonctions adaptées (harmoniques toroïdales) fournit un critère permettant de qualifier les estimations de la frontière plasma. Un algorithme de descente permet de le faire décroître, en améliorant l'estimation de la frontière. Cette méthode, qui ne requiert pas d'intégration de l'équation dans le domaine, fournit de très bons résultats et semble appelée à connaître des extensions pour d'autres tokamaks tels que JET et ITER.