Malgré la complexité du processus de transformation, plusieurs systèmes modèles ont été développés dans lesquels les cellules normales se transforment d'une manière progressive par l'introduction d'éléments génétique. Ici, seules les cellules transformées rentrent en apoptose induite par TRAIL. Comme le milieu des cellules sénescentes a déjà été impliqué dans certaines caractéristiques tumorales (la prolifération, l'invasion et d'autres), notre objectif était d'évaluer soneffet potentiel sur l'acquisition de la sensibilité à TRAIL. Lorsque toutes les cellules de ce système de transformation sont incubés avec le milieu conditionné de cellules sénescentes (CMS) la sensibilisation à TRAIL a été observée dans les cellules pré-transformées mais pas dans les cellules immortalisées ou normales. Ainsi, nous avons conclu que les différentes étapes de la transformation fournissent un contexte cellulaire et moléculaire particulier qui agit en synergie avec le CMS afin de promouvoir la sensibilisation à TRAIL. Ces observations mettent l'accent sur le rôle spécifique descellules sénescentes et leur phénotype sécrétoire. Nous avons ensuite étudié les mécanismes activés dans les cellules pré-transformées par le CMS et responsables de leur sensibilisation à TRAIL. Nos résultats suggèrent un rôle clé de l'axe Myc-FLIPL dans la signalisation activée par le CMS.

The cytokine TRAIL is a promising cancer therapeutic candidate as it induces apoptosis selectively in transformed cells. TRAIL-induced clustering of its receptors (DR) is essential for the DISC complex formation, which induces cell death. The mechanism for TRAIL's tumour selective effect is largely unknown. We identified the cytoskeleton proteins non-muscle myosin heavy chain IIa, IIb (NMHCIIa, NMHCIIb), myosin regulatory light chain (MLC2) and ß-actin as novel DR-interactors. An initially weak and TRAIL-induced abrogation of NMHCII/DR interaction correlated with efficient DISC formation in tumour cells. In contrast, a robust NMHCII/DR interaction that was sustained upon TRAIL stimulus was accompanied by incomplete DISC arrangement. Weakening the NMHCII/DR interaction in normal cells using chemical inhibitors enhanced TRAIL-induced apoptosis. Intriguingly, siRNA-mediated NMHCIIa- but not NMHCIIb depletion potently released TRAIL resistance in normal cells and influenced DISC composition. Reduced NMHCII/DR interaction in transformed cells was characterised by diminished MLC2 phosphorylation and altered protein expression of upstream regulatory kinases. Our results suggest that normal cell resistance to TRAIL-apoptosis is based on the interaction of cytoskeleton components with DR that is impaired upon transformation. Since NMHCII function in cell adhesion and migration, it will be interesting to study possible roles of the interaction in cell detachment and altered TRAIL sensitivity; moreover this link may provide clues as to the cause of TRAIL resistance in some cancers.

L'objectif de ce travail de thèse a été d'analyser les stratégies de réduction de la fatigue musculaire en course de trail et potentiellement d'identifier certains paramètres d'influence de cette fatigue. La course de trail est un nouveau sport en essor qui induit une combinaison spécifique de fatigue et dommages musculaires des principaux muscles locomoteurs. Afin de pouvoir conduire des études interventionnelles, une étude descriptive préliminaire a été conduite pour caractériser la fatigue spécifique et les dommages musculaires induits par ce type d'épreuve de trail. Ensuite, la reproductibilité du trail comme modèle de fatigue a été vérifiée afin de pouvoir l'utiliser dans un contexte d'intervention. Enfin, deux études visant à réduire la fatigue induite par le trail ont été conduites. D'une part l'utilisation des vêtements de compression - très à la mode en trail a été analysée comme stratégie d'optimisation de la performance. D'autre part, a aussi été étudié l'effet d'un réchauffement préalable du muscle sur les dommages musculaires : Dans cette optique, une étude contrôlée en laboratoire a été menée, examinant les effets d'un réchauffement passif sur les conséquences fonctionnelles de course en descente chez une population non-entraînée. En résumé, les travaux conduits au sein de cette thèse fournissent une description de la fatigue en trail, et valident l'utilisation du trail comme modèle reproductible de terrain pour investiguer les stratégies de réduction de la fatigue. De plus, ils relativisent l'effet positif des vêtements de compression sur la performance et montrent le lien fonctionnel entre le réchauffement musculaire et la réduction des dommages musculaires induits par un travail excentrique.

The overall aim of this thesis is to investigate if the somatic comorbidity and increased mortality among persons with psychiatric disability (residents in supported housing facilities) can be influenced. The thesis comprises four papers. Paper I describes the lived experience of health and body. Papers II and III examine the effects of a lifestyle programme on physiological markers (II) and on psychological and quality of life parameters (III). Paper (IV) illustrates the meaning of participating in a life style programme. The data in Papers I and IV comprises narrative interviews with residents (n=11). The studies in Papers II and III are focused on residents and were carried out with a randomized design. The randomization was performed on a group level (supported housing facility). The 12 month intervention consisted of study circles with a theoretical and practical application of dietary information and physical activity for two hours, on a twice weekly basis under the supervision of a study circle leader. The controls were offered an aesthetic study circle and met once a week. The data in Paper II comprises physiological quantitative data from both residents (n=41) and staff (n=41) and in Paper III questionnaires on symptoms and quality of life completed only by residents (n=41). The data was analysed with Qualitative description (I), Phenomenological-hermeneutics (IV), and for papers II and III relevant statistical calculations were used. Health is described in paper I as “having a life as others have” and discloses the losses of important life domains (family, work, security) and the experiences of being deviant and stigmatized. Health is described as “absence of psychological and physical problems” and its hampering effects on quality of life and self-esteem. Health is understood as a phenomenon that could “be influenced by one self”, and there is an insight that health is manageable. Participating in a life style intervention (paper II) meant a significant improvement in risk factors for metabolic syndrome among the residents in comparison with controls. No differences were seen on weight, BMI and improved physical capacity. In paper III a significant positive increase in the Sense of Coherence compared to controls was seen. However no effect was seen on quality of life, psychosocial function or on reduction of symptoms in comparison with controls. Participating in a lifestyle intervention can be understood as the gaining of insights that health can be improved and that the daily life is partly given a changed content (paper II). The participation is also described as meaning an increased sense of closeness and equality in relation to the staff and sometimes a painful insight of their life situation. Participating is also described as entailing a hope that one’s life situation can be affected. In summary this thesis shows that there is some possibility of influencing the physical health (reduced risk of metabolic syndrome) among persons with psychiatric disability by participating in a 12 month intervention programme. The intervention does not show any effects on measures such as quality of life, psychosocial function and presence of symptoms. However, the participants describe that the participation had a meaning in a number of respects. This is a finding that is confirmed by the positive change in sense of coherence. The need to develop preventive care for persons with psychiatric disability and the importance of monitoring the treatment with neuroleptics and its side effects on physical health is an important clinical implication. Furthermore the importance of the responsibility of the care staff is emphasized as well as the importance of supporting a change in lifestyle.

As the main goal of this thesis was determined a project proposal for optimization of educational trails in the selected area of the National park and Protected landscape area Šumava. Purpose of the project is the improvement of tourism by creating a new product. Based on the analysis the primary data was designed optimization project:Educational cycling route. The intention is to link the major attractions of the Šumava´s foothills Prachatice with cultural and natural attractions of Volary, which is situated in a protected landscape. The purpose of the project is to introduce visitors with the landscape and history and promote their awareness of environmental protection for a non-violent way. For the project was defined technical equipment, furniture routes. On the basis of delivering of tangible assets were determined the total costs.

The objective of this diploma thesis is search processing of tourism, his problematics and protection of the nature. In the other part this thesis deal with analysis of hiking trail in Labske piskovce PLA in focus of potentional atractive with proposal of new hiking trails in this location. Analysis containe all hiking trails in this location according to colored tourism distinction of trails. Proposal of new hiking trails was formed with regard on protection of nature thus at the same time was surrounding area approached for visitors as much as possible. Proposal comprise new and turisty often attended attractiviness. This proposal can produce increase of attractiviness in this district and the influx of new turists.

Proteomics is a set of analytical methods which enable qualitative and quantitative characterization of the proteome. Expression proteomics quantitatively compares proteomes of cells, tissues, body fluids or other biological materials to find differencies in protein expression and, based on these differencies, to describe the biological processes occuring in investigated organisms. An initial material for expression proteomic studies are complex mixtures containing thousands of proteins, which are analyzed using separation (electrophoretic and chromatographic) methods, and identified, possibly quantified using mass spectrometry. The aim of this Thesis is to demonstrate the application of the tools of expression proteomics in solving diverse challenges in biomedicine. We employed various proteomic approaches and tools for studying molecular mechanisms of human diseases using pacient biological samples, or a model organism and a cell culture. We were conducting three different research projects, namely: A quest for potencial molecular targets for selective elimination of TRAIL-resistant mantle cell lymphoma cells; Investigation of molecular mechanisms of heart failure using a rat model of the disease induced by volume overload; and Searching for diagnostically usable serum biomarkers of ovarian...

The present diploma paper discusses the locality of Borkovice marshes. It consists of two parts, a theoretical part and a practical part, the theoretical part mainly imparting theoretical information about the region, its communities and subventions and the practical part dra-wing attention to the locality, its specificities and its tourist attractions. Furthermore, with the help of SWOT analysis, the paper postulates the possibilities of region's development. One of the steps taken to aid the region and described in the present paper was the creation of the Legal Association of Blata municipality. In conclusion, a draft has been included of an application for a subvention, to be submitted by the Legal Association to the region of Southern Bohemia.

O efeito ansiolítico dos benzodiazepínicos (BZDs) é reduzido depois da primeira exposição ao labirinto em cruz elevado (LCE). Várias hipóteses tem sido formuladas para explicar este fenômeno chamado one-trial tolerance (OTT), entretanto, nenhuma delas é conclusiva. No presente estudo, examinamos este fenômeno através da análise etofarmacológica de ratos submetidos ao LCE em duas sessões (T1 e T2), e do conteúdo de monoaminas presentes no córtex pré-frontal, amígdala, hipocampo e núcleo accumbens através da técnica de Cromatografia Líquida de Alta Pressão. Ratos machos Wistar foram tratados com salina ou midazolam (0,5 mg/Kg, i.p.) antes de T1 e T2 e imediatamente depois, seus encéfalos foram dissecados e as estruturas analisadas. Como controle à análise neuroquímica foram incluídos animais tratados com salina e não expostos ao LCE. A administração de midazolam antes de T1 promoveu efeito ansiolítico, aumentando a exploração dos braços abertos, porcentagem de entradas e tempo de permanência nos mesmos. Em T2 foi observado redução da exploração dos braços abertos em relação a T1. Esses resultados sugerem que há uma mudança no estado emocional do animal em T2, que é resistente a ação ansiolítica dos BZDs. Com relação aos resultados dos estudos neuroquímicos, foi observado redução dos conteúdos de serotonina (5- HT) e noradrenalina (NA) no córtex pré-frontal, na amígdala, no hipocampo e no núcleo accumbens depois de T1 e T2. Houve também, redução do conteúdo de dopamina (DA) na amígdala depois de ambas sessões. Não ocorreram mudanças nas taxas de renovação dessas monoaminas em nenhuma das estruturas analisadas. Através desses resultados, pode-se inferir que a estimulação aversiva do LCE causa alterações na neurotransmissão monoaminérgica da amígdala, como também das outras estruturas límbicas estudadas neste trabalho. Essas alterações neuroquímicas depois da primeira exposição ao LCE, devem representar alterações adaptativas na neurotransmissão do sistema límbico que podem estar associadas ao fenômeno da OTT. / Numerous reports have demonstrated that a single exposure to a variety of stressful experiences enhances fearful reactions when behavior is subsequently tested in current animal models of anxiety. Until now, no study has examined the neurochemical changes during the test and retest sessions of freely-behaving rats in the elevated plus-maze (EPM), one of the most traditional tests of anxiety. This work is a new approach looking at the changes in dopamine (DA), serotonin (5-HT) and noradrenaline (NA) levels in the prefrontal cortex, amygdala, hippocampus and nucleus accumbens during one-trial learning development. We used high pressure liquid chromatography to assess the concentrations of these neurotransmitters and their metabolites in animals injected with saline or midazolam upon single or double exposure to the EPM. For the biochemical analysis an extra control group treated with saline and not exposed to EPM was added. The data showed that stressful stimuli present in the maze were able to elicit one-trial learning to midazolam on re-exposure. Significant decreases in 5-HT and NA contents in the prefrontal cortex, amygdala, hippocampus and nucleus accumbens occurred in saline and midazolam injected animals submitted to the first and second trials. Significant decreases in DA content were also observed in the amygdala after both trials. There was no change in the turnover of these monoamines in any structure studied. It is suggested that aversive stimuli inherent to the EPM cause primary changes in the neurochemical mechanisms of the amygdala and also influence the activity of monoaminergic neurotransmission in the prefrontal cortex, hippocampus and nucleus accumbens. The observed reduction in monoaminergic transmission in limbic structures after the first stressful experience in the EPM seems to represent adaptive changes and may be associated to the phenomenon of ?one-trial tolerance?.

The 85th percentile off-road glances while performing three tasks on an in-vehicle system while driving in a simulator was investigated. The tasks were a radio task, a telephone task and a sound settings task which were performed at three occasions each. The distribution of 85th percentile off-road glance durations for each subject and task showed that durations differed between individuals rather than between tasks. It also turned out that durations longer than 2.00 seconds were not rare and 2 of 16 subjects had durations longer than 2.00 seconds in the radio task. Even though the distribution showed small differences between tasks on an individual level, differences on a group level were found between the tasks. A tendency of a learning effect was found, which implied a decrease in 85th percentile off-road glance durations as the tasks were performed at several occasions. A tendency of a floor effect in 85th percentile off-road glance durations, when the subjects are familiarized with tasks, was also found. Performance on a computerized trail-making test, measuring ability of visual search, motor speed and mental flexibility, was found not to be related with 85th percentile off-road glance durations.