The Diplom's Thesis "Mariology of Louise M. Grignion de Montfort and Leo XIII" The Marian worshiper Louis Maria Grignion of Montfort worked in France as a Catholic priest and missionary at the turn of the 17th and 18th centuries. Grignion, the promoter of the rosary prayer and marian veneration, which Pope Leo XIII blessed in 1888, on the 50th anniversary of his ordination to the priesthood, left in his writings a spiritual message and taching, the elements of which we find in the works of Pope Leo XIII. This Pope is the author of Marian encyclicals promoting the rosary prayer. The tematic similarity in his view of the figure of the Virgin Mary and her influence in history appears in both, Grignion and Leo XIII, in the field of mariological expression and in the inseparable trinitological basis of mariology. The first chapter of this work contains the biography of the priest and missionary Louis M. Grignion of Montfort. It is followed by a biography of Gioacchino Pecci, the pope of the Roman Catholic Church since 1887. The second chapter clarifies the periodic conditionality of Grignion's works and outlines his partial mariology and Marian veneration. The third chapter maps the thinking of Louis M. Grignion implicitly present in the Marian encyclicals of Leo XIII on a general level and then in the...

Driving comfort is considered a key factor for broad public acceptance of automated driving. Based on continuous driver/passenger monitoring, potential discomfort could be avoided by adapting automation features such as the driving style. The EU-project MEDIATOR (mediatorproject.eu) aims at developing a mediating system in automated vehicles by constantly evaluating the performance of driver and automation. As facial expressions could be an indicator of discomfort, a driving simulator study has been carried out to investigate this relationship. A total of 41 participants experienced three potentially uncomfortable automated approach situations to a truck driving ahead. The face video of four cameras was analyzed with the Visage facial feature detection and face analysis software, extracting 23 Action Units (AUs). Situation-specific effects showed that the eyes were kept open and eye blinks were reduced (AU43). Inner brows (AU1) as well as upper lids (AU5) raised, indicating surprise. Lips were pressed (AU24) and stretched (AU20) as sign for tension. Overall, facial expression analysis could contribute to detect discomfort in automated driving.

La voie de réparation par excision de nucléotides (NER) répare les lésions qui distordent la double hélice d’ADN notamment ceux induits par l’irradiation UV. Le NER est subdivisé en deux sous-voies : GG-NER (Global Genome Repair) et TC-NER (Transcription-Coupled Repair). La sous-voie GG-NER enlève les dommages à l’ADN dans l’ensemble du génome. La sous-voie TC-NER répare les dommages sur le brin transcrit qui interfèrent avec la progression de l’ARN Pol II. Les défauts de la voie NER peuvent conduire à l’apparition de pathologies graves. Par exemple, des mutations dans le gène XPG, codant une 3’ endonucléase impliquée dans la voie NER, peuvent mener au xeroderma pigmentosum (XP) associé ou non au syndrome de Cockayne (CS).Récemment, le laboratoire a découvert un lien fonctionnel entre Rad2, homologue chez la levure Saccharomyces cerevisiae de la protéine XPG humaine, et le Médiateur (Eyboulet et al., 2013). Le Médiateur est un complexe multiprotéique nécessaire à la régulation de la transcription dépendante de l’ARN Pol II. Cette étude a suggéré que le Médiateur est impliqué dans la sous-voie TC-NER en facilitant le recrutement de Rad2 au niveau des régions transcrites.Mon projet de thèse visait à étudier les mécanismes moléculaires qui lient la transcription et la réparation de l’ADN. Plus précisément, d’investiguer le lien fonctionnel entre le Médiateur et la machinerie du NER chez S. cerevisiae.Lors du TC-NER, l’ARN Pol II est le premier facteur signalant le dommage à l’ADN. De plus, le Médiateur et Rad2 interagissent avec l’ARN Pol II. Pour déterminer le lien fonctionnel entre ces composants, nous avons utilisé des approches de génétique et génomique dans les mutants de TFIIH (kin28), de l’ARN Pol II (rpb9) and du Médiateur (med17). Nos résultats nous ont permis de proposer un modèle dans lequel Rad2 est recruté au niveau des régions régulatrices enrichies par le Médiateur, et Rad2 est ensuite transféré au niveau des régions transcrites de manière dépendante à l’ARN Pol II. De plus, ces résultats suggèrent que le rôle du Médiateur dans la transcription est fortement lié à son rôle dans la réparation de l’ADN.Ensuite, nous avons montré que le lien entre le Médiateur et la machinerie du NER peut être étendu à d’autres protéines du NER notamment en démontrant une interaction physique entre le Médiateur et Rad1/XPF, Rad10/ERCC1 ou Rad26/CSB, en l’absence des UV. Tout comme Rad2, nous avons démontré que Rad1 et Rad10 n’ont pas de rôle majeur dans la transcription. Pour approfondir le lien entre ces protéines du NER et le Médiateur, des expériences de ChIP-sequencing ont été réalisées. Nous avons observé que le Médiateur est présent au niveau de certaines régions qui sont aussi enrichies par ces protéines du NER. Après l’induction des dommages par UV, les interactions entre le Médiateur et la machinerie du NER reste inchangées par rapport aux conditions en l’absence des UV. De plus grâce à nos expériences de ChIP, nous avons observé un changement de la liaison à la chromatine des protéines du NER et du Médiateur après l’irradiation aux UV. Des expériences de ChIP-sequencing seront réalisées pour avoir une vue globale de ces changements.En conclusion, nous avons solidifié le lien fonctionnel entre Rad2, le Médiateur et l’ARN Pol II par rapport à la réparation couplée à la transcription. Nous avons aussi démontré que le Médiateur interagit avec d’autres protéines du NER (Rad1/XPF, Rad10/ERCC1 et Rad26/CSB) et colocalise avec eux sur certaines régions de la chromatine. En somme, notre projet place le Médiateur à l’interface de la transcription et de la réparation de l’ADN, deux processus essentiels dont les défauts peuvent mener à des pathologies graves. / Nucleotide excision repair (NER) is a well conserved pathway that removes helix-distorting DNA lesions such as those arising upon UV irradiation. Global genome repair subpathway (GG-NER) removes the DNA lesions in the genome overall, and transcription-coupled repair (TC-NER) removes the DNA lesions interfering with Pol II progression through actively-transcribed regions. Defects in the NER pathway may lead to severe human pathologies. For instance, mutations in human XPG gene, encoding a 3’ endonuclease essential for NER, give rise to xeroderma pigmentosum (XP) sometimes associated with Cockayne syndrome (CS). Recently, the laboratory discovered a functional link between Rad2/XPG and Mediator in Saccharomyces cerevisiae (Eyboulet et al., 2013). Mediator is a large multisubunit complex essential for transcription regulation. We suggest that Mediator is involved in TC-NER by facilitating Rad2 recruitment to transcribed genes.My PhD work aimed at addressing the molecular mechanisms of this link between transcription and DNA repair, especially by investigating the functional interplay between Mediator and the NER machinery in yeast Saccharomyces cerevisiae.RNA Pol II is the first complex of TC-NER that encounters the DNA damage. Moreover, both Mediator and Rad2/XPG interact with Pol II. However, a functional interplay between all these components related to TC-NER remained to be determined. Using genetic and genomic approaches, in particular ChIP-sequencing in TFIIH (kin28), RNA Pol II (rpb9) and Mediator (med17) mutants, our work led us to propose a model where Rad2 shuttles between Mediator on upstream activating sequence (UAS) and RNA Pol II on transcribed regions (Georges, Gopaul et al., under review). Our results also suggest that Mediator functions in transcription and DNA repair are closely related.Moreover, we showed that Mediator’s link to NER can be extended to other NER proteins. Indeed, we identified a physical interaction between Mediator and other NER proteins, including Rad1/XPF, Rad10/ERCC1 and Rad26/CSB in the absence of UV irradiation. Similarly to Rad2, we demonstrated that Rad1 and Rad10 do not have a major role in yeast transcription. To further study the functional link between Mediator and the NER machinery, we obtained the genomic distribution of different NER proteins by ChIP-sequencing. We found that some promoter regions are co-occupied by Mediator and these NER proteins, and that relationships between Mediator and these NER proteins are more complex than between Mediator and Rad2. We also investigated if physical interactions between Mediator and NER proteins are modified after UV, we did not observe any significant change. Furthermore, we observed that the chromatin binding profiles of NER proteins and Mediator are modified after UV-irradiation. ChIP-sequencing will be carried out to get a genome-wide view of their chromatin binding profiles.In conclusion, we have strengthened the link between Rad2/XPG, Mediator and RNA Pol II, providing mechanistic insights into functional interplay between these components related to transcription-coupled repair, and showed that the link between Mediator and the NER machinery can be extended to other proteins. Taken together, our results suggest a close relation between Mediator functions in transcription and in NER, two fundamental processes dysfunction of which leads to human diseases.

The thesis focuses on the role of an adult-parents in the viewing and reading of a series of picture-free text books (so-called wimmelbuch) by S. Bernerová  Winter, Spring, Summer, Autumn, Night. The aim of the thesis is to describe what interactions occur between a parent and a child between two and three years of age when reading "picture books", so-called wimmelbuch. The theoretical part of the thesis deals with the topics of reading literacy development (especially for two-year and three-year-olds), pictorial books, books of the type wimmelbuch, adult as a children's book mediator, illustrations and reading rituals. The empirical part includes quantitative and qualitative research. The quantitative part of the research is based on data from an online questionnaire for parents of preschool children. In the framework of long-term qualitative research, the interaction between parent and child between two and three years of age in four selected families is monitored and described, while working with books of the type wimmelbuch by Susanne Berner. The result of the questionnaire is that more than half of the parents interviewed have some of books of the type wimmelbuch by S. Berner, and most of them think that these books are developing children's reading literacy. Most often, viewing/reading takes...

The thesis deals with the question of Egypt's role of a biased mediator towards the Palestinian side in the Israeli-Palestinian conflict, and its stabilising effects on the region in the form of a case study applying the theory of a biased mediator in conflict resolution on the Egyptian example. In addition to this, the goal of this thesis is to demonstrate the beneficial effects of different forms of mediation activities and their combination, including biased and neutral mediation, in the transformation of intractable and protracted conflicts into their tractable form, using the example of the Israeli-Palestinian conflict. The identification of Egypt's positive role in contributing to the resolving of the Israeli-Palestinian conflict, of which it was once a warrying party, portrays the transformational capacity of conflicts. The thesis also highlights Egypt's potential in moderating extremist trends in the region, also through the successful mediation efforts in the inter-palestinian conflict involving the Fatah and Hamas movements, contributed to by Egypt's changing position in the region, her historical role in the Israeli-Palestinian conflict and her strategic and security concerns in the region.

OVERVIEW This study deals with the situation of mediation in the Czech Republic. Mediation began to develop as a modern field during the 1990s. With the adopting of Act No. 202/2012 Coll. on Mediation, partial definitions were give for what mediation is and for the conditions under which it can be provided. Nonetheless, the situation in mediation is still very complicated. The study therefore presents a unified overview of what mediation is, of the principles upon which it is based, and of how it is used in practice. It introduces the role of a mediator and of the demands that must be satisfied for the performance of mediation, and it describes how the professional standards and code of ethics for mediators are established. By comparing the profession of an attorney to that of a mediator on the basis of the identifying characteristics of an expert profession, it determines whether mediation is a separate profession or a method that is useful in other fields.

Indiana University-Purdue University Indianapolis (IUPUI) / The voltage-gated sodium channel Nav1.7 has gained traction as a pain target with recognition that loss-of-function mutations in SCN9A, the gene encoding Nav1.7, are associated with congenital insensitivity to pain, whereas gain-of-function mutations produce distinct pain syndromes due to increased Nav1.7 activity. Selective inhibition of Nav1.7 is fundamental to modulating pain via this channel. Understanding the regulation of Nav1.7 at the cellular and molecular level is critical for advancing better therapeutics for pain. Although trafficking of Nav1.7 remains poorly understood, recent studies have begun to investigate post-translational modifications of Navs and/or auxiliary subunits as well as protein-protein interactions as Nav-trafficking mechanisms. Here, I tested if post-translational modifications of a novel Nav1.7-interacting protein, the axonal collapsin response mediator protein 2 (CRMP2) by small ubiquitin-like modifier (SUMO) and phosphorylation could affect Nav trafficking and function. Expression of a CRMP2 SUMOylation incompetent mutant (CRMP2-K374A) in neuronal model CAD cells, which express predominantly Nav1.7 currents, led to a significant reduction in huwentoxin-IV-sensitive Nav1.7 currents. Increasing deSUMOylation with sentrin/SUMO-specific protease SENP1 or SENP2 in wildtype CRMP2-expressing CAD cells decreased Nav1.7 currents. Consistent with reduced current density, biotinylation revealed significant reduction in surface Nav1.7 levels of CAD cells expressing CRMP2-K374A or SENP proteins. Diminution of Nav1.7 sodium current was recapitulated in sensory neurons expressing CRMP2-K374A. Because CRMP2 functions are regulated by its phosphorylation state, I next investigated possible interplay between phosphorylation and SUMOylation of CRMP2 on Nav1.7. Phosphorylation of CRMP2 by cyclin dependent kinase 5 (Cdk5) was necessary for maintaining Nav1.7 surface expression and current density whereas phosphorylation by Fyn kinase reduced CRMP2 SUMOylation and Nav1.7 current density. Binding to Nav1.7 was decreased following (i) loss of CRMP2 SUMOylation, (ii) loss of CRMP2 phosphorylation by Cdk5, or (iii) gain of CRMP2 phosphorylation by Fyn. Altering CRMP2 modification events simultaneously was not synergistic in reducing Nav1.7 currents, suggesting that Nav1.7 co-opts multiple CRMP2 modifications for regulatory control of this channel. Loss of either CRMP2 SUMOylation or Cdk5 phosphorylation triggered Nav1.7 internalization involving E3 ubiquitin ligase Nedd4-2 as well as endocytosis adaptor proteins Numb and Eps15. Collectively, my findings identify a novel mechanism for regulation of Nav1.7.

Indiana University-Purdue University Indianapolis (IUPUI) / Huntington’s Disease (HD) is an inherited, autosomal dominant, neurodegenerative disorder. There is no cure for HD and the existing therapies only alleviate HD symptoms without eliminating the cause of this neuropathology. HD is linked to a mutation in the huntingtin gene, which results in an elongation of the poly-glutamine stretch in the huntingtin protein (Htt). A major hypothesis is that mutant Htt (mHtt) leads to aberrant Ca2+ homeostasis in affected neurons. This may be caused by increased Ca2+ influx into the cell via the N-methyl-Daspartate (NMDA)-subtype of glutamate receptors. The contribution of two major Ca2+ removal mechanisms, mitochondria and plasmalemmal Na+/Ca2+ exchangers (NCX), in neuronal injury in HD remains unclear. We investigated Ca2+ uptake capacity in isolated synaptic (neuronal) and nonsynaptic mitochondria from the YAC128 mouse model of HD. We found that both Htt and mHtt bind to brain mitochondria and the amount of mitochondriabound mHtt correlates with increased mitochondrial Ca2+ uptake capacity. Mitochondrial Ca2+ accumulation was not impaired in striatal neurons from YAC128 mice. We also found that expression of the NCX1 isoform is increased with age in striatum from YAC128 mice compared to striatum from wild-type mice. Interestingly, mHtt and Htt bind to the NCX3 isoform but not to NCX1. NCX3 expression remains unchanged. To further investigate Ca2+ homeostasis modulation, we examined the role of collapsin response mediator protein 2 (CRMP2) in wild-type neurons. CRMP2 is viewed as an axon guidance protein, but has been found to be involved in Ca2+ signaling. We found that CRMP2 interacts with NMDA receptors (NMDAR) and disrupting this interaction decreases NMDAR activity. CRMP2 also interacts with and regulates NCX3, resulting in NCX3 internalization and decreased activity. Augmented mitochondrial Ca2+ uptake capacity and an increased expression of NCX1 in the presence of mHtt suggest a compensatory reaction in response to increased Ca2+ influx into the cell. The role of NCX warrants further investigation in HD. The novel interactions of CRMP2 with NMDAR and NCX3 provide additional insight into the complexity of Ca2+ homeostasis regulation in neurons and may also be important in HD neuropathology.

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5 Abstract Genetic mechanisms of regulation of gene expression form the basis for proper development, function of organisms and their responses to variable life conditions. However, they are relatively slow. Life processes that require a fast response to the changing environmental and metabolic conditions are mostly executed on the level of proteins especially their posttranslational modifications and protein- protein interactions.The goal of the experimental work that led to the presented thesis consisted in exploitation of the model organism Caenorhabditis elegans for analysis of regulation of gene expression by transcription factors from the protein family of nuclear receptors. The model system C. elegans enables very efficient experimental procedures in the field of genetics, genomics and functional analysis of phenotypes. In the experimental work connected with this thesis, I studied the regulation of gene expression under specific experimental conditions from the perspective of advanced functional proteomics and I focused on the employment of separation methods and methods of advanced proteomics, especially by mass spectrometry.In the first part of the work, I characterized the nuclear receptor NHR-60 on the protein level. This nuclear receptor is expressed as two protein forms with a mass of 50 kDa...