The Cooperative Game Theory is a scientific discipline which offers rich mathematical apparatus for describing complex situations in the social reality. Its apparatus includes an extension to hierarchical structures and therefore can be applied to numerous research problems from the International Relations field. However, a cooperative game theoretical approach is very scarcely used. The main goal of the diploma thesis is to demonstrate, on the research problem of decision making in participation in local conflicts, the benefits of results that can be achieved by the application of the Cooperative Game Theory. In the first part of the thesis, theoretical foundations are laid and basic concepts are introduced. The second part is focused on forming a series of models of cooperative games with hierarchical structures from four local conflict situations, which are subsequently restricted in order to describe authoritative relations in structure. Restricted games are solved, the results are interpreted and evaluation of how these results can contribute to addressing the research problem follows.

Les protéines IFITM (« InterFeron Inducible TransMembrane proteins »), et en particulier les membres 1, 2 et 3, sont des facteurs de restriction antiviraux dont l’expression est induite par le système immunitaire inné en réponse à une infection virale. Elles inhibent la réplication de nombreux virus pathogènes pour l’homme parmi lesquels figurent le virus de la grippe A, le VIH (Virus de l’Immunodéficience Humaine) de type 1 ou encore le virus de l’hépatite C. Ces virus entrent dans la cellule hôte, soit par fusion directe avec la membrane plasmique, soit par la voie de l’endocytose. Il est à présent communément admis que les protéines IFITM, localisées au sein des membranes plasmiques et endolysosomales, agissent en inhibant la fusion des membranes virales et cellulaires, empêchant par conséquent l’entrée du virus dans la cellule et donc sa réplication. D’autre part, dans le cas du VIH, leur incorporation dans les particules virales produites par la cellule hôte diminuerait la capacité de ces particules à infecter de nouvelles cellules cibles. Cependant, les mécanismes moléculaires par lesquels les protéines IFITM interfèrent avec le cycle viral ne sont pas encore clairement définis.Parmi les membres de la famille IFITM, IFITM3 est celui qui présente l’effet antiviral le plus systématique selon les différentes études. Il constitue donc un modèle de référence pour étudier la famille IFITM.Déterminer la structure ainsi que la topologie membranaire d’IFITM3 sous sa forme active rendrait alors possible la réalisation d’études fonctionnelles, dont les résultats contribueraient sans nul doute à élucider le(s) mécanisme(s) par le(s)quel(s) IFITM3 exerce son activité antivirale.C’est pourquoi, nous nous sommes tout d’abord attelés à reconstituer IFITM3 au sein de membranes artificielles (liposomes, nanodisques), car contrairement aux micelles de détergent, ces membranes artificielles peuvent mimer l’environnement natif des protéines membranaires et par conséquent, offrir de plus grandes chances de les y étudier sous leur forme active. Nous avons ensuite procédé à la caractérisation biochimique et biophysique d’IFITM3 et avons mis en évidence la formation de dimère de la protéine ainsi que de plus grandes espèces oligomériques. L’analyse structurale d’IFITM3 reconstituée en nanodisques par RMN nous a quant à elle permis d’identifier une courte région hélicoïdale dans la région N-terminale extramembranaire d’IFITM3 encore jamais décrite auparavant et pouvant correspondre à un motif d’internalisation. Nous avons en outre observé, par microscopie électronique à coloration négative, de potentiels effets d’IFITM3 sur la courbure de la membrane de liposomes qui pourraient être à l’origine de son action inhibitrice sur la fusion virale. Et enfin, nous avons montré au travers d’expériences TEVC que lorsqu’IFITM3 est présente dans l’environnement extracellulaire d’ovocytes de xénope, celle-ci est capable d’engendrer des fuites ioniques au travers de la membrane des ovocytes qui pourraient résulter soit, d’une déstabilisation de la membrane par IFITM3 soit, d’une formation de pores membranaires par la protéine. / The host cell first line of defence against viral infections induces the production of interferons. These interferons are then released in the surrounding medium where they bind to target cells and induce the expression of hundreds of genes so called interferon-stimulated genes (ISGs). The interferon inducible transmembrane proteins IFITM are part of the products of these ISGs. IFITM1, 2 and 3 are antiviral factors able to restrict the replication of a broad variety of enveloped viruses, such as influenza virus, HIV-1 (Human Immunodeficiency Virus) and Hepatitis C virus. These viruses enter in the host cell either by direct fusion with the cell membrane or by endocytosis. IFITM proteins contain two membrane regions for insertion or interaction with plasma and endolysosomal membranes where they block the fusion of virus particles with cellular membranes by a mechanism which is still undefined. In addition, their incorporation into new HIV virions, in virus producing cells, has been correlated with decreased infectivity.Among the IFITM protein family members, IFITM3 is the one showing the most recurrent antiviral effect in the different studies. Therefore it represents a good model to study the whole IFITM family.The determination of its structure and membrane topology is crucial to be able to clarify, through structure-based functional studies, the mechanism(s) by which IFITM3 interfere with the viral cycle.Here we characterized and we studied IFITM3 structure and membrane topology in a lipidic environment close to its native environment such as liposomes and nanodiscs. We demonstrated that IFITM3 can self-associate to form at least a dimer. Some higher order associations of IFITM3 have been observed after its reconstitution into liposomes and big size nanodiscs. We discovered by NMR in solution that the N-terminal region of IFITM3 contains a small helical region, never described until now, which could correspond to an internalization motif. We also observed by negative staining electron microscopy some liposomal membrane curvature changes that could be assigned to the presence of IFITM3 in these liposomes. And we discovered through TEVC experiments that IFITM3 addition in the extracellular environment of xenopus oocytes produces ion leaks through the oocyte membrane which could result either from membrane destabilization or from a pore formation.

Restriction of legal capacity proceedings Abstract This diploma thesis deals with the legal regulation of legal capacity proceedings. Integral part of this thesis is also analysis of related legal capacity substantive law and also other supportive measures such as preliminary declaration, representation of household member and assistance contract. The procedure which leads either to restriction of legal capacity, time extension, change or cancellation of the restriction is however the main point of this thesis. The goal of this thesis is to provide comprehensive view on this type of proceedings in the context of relevant case law and courts practice. Thesis also shows the practical use of this institute in Czech Republic. It also contains the issue of mental illness and its impact on the daily life, the number of persons restricted in their legal capacity, the lenght of court proceedings and the issue of ID cards not containing the information about restricted legal capacity and its possible associated negative consequences. The author tries to suggest possible solutions to the problematic aspects of the legislation and current praxes. The thesis is divided into four chapters. In the first chapter, the author writes about the possible alternatives the legal regulation offers to people who are not fully able...

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Bakgrund: Övervikt och fetma ökar exponentiellt i alla samhällsklasser i alla delar av världen idag. Det kan orsaka lidande och förtida död för de drabbade. Risken att drabbas av våra vanligaste vällevnadssjukdomar som hjärt- och kärlsjukdom, typ 2 diabetes och neurodegenerativa sjukdomar som Alzheimers sjukdom ökar vid övervikt och fetma. Det i särklass mest effektiva sättet att gå ned i vikt är genom att minska sitt kaloriintag. För att behålla en viktnedgång behövs beteendeförändringar, stöttning och en långsiktig plan. Syfte: Syftet med studien är att undersöka om intermittent kalorirestriktion (CR) är mer effektivt än kontinuerlig CR när det gäller viktreduktion och bibehållande av ny vikt. Metod: En litteraturstudie genomfördes där fyra olika parallellstudiers resultat sammanställs. Parallellstudierna jämför effektiviteten av intermittent och kontinuerlig CR på viktreduktion och bibehållande av ny vikt. Resultat: I tre av de fyra studier som undersökts kan inga skillnader i effektivitet uppvisas mellan intermittent och kontinuerlig CR när det gäller viktreduktion och bibehållande av ny vikt över tid. En av studierna fann att intermittent CR, i form av ”week on, week off” två veckor i taget är mer effektiv när det gäller viktreduktion efter 16 veckors interventionsfas samt 6 månader senare. Slutsats: Som alla metastudier jag läst och de flesta studier på ämnet kan ingen skillnad i effektivitet mellan intermittent och kontinuerlig CR uppvisas gällande viktreduktion. Inte heller verkar den ena vara överlägsen den andra när det gäller bibehållande av ny vikt. Fler studier över längre tidsspann med fler deltagare krävs för att kunna dra slutsatser om vilken typ av CR som är mest effektiv på lång sikt. / Background: Overweight and obesity are increasing exponentially in all social classes in all parts of the world today. It can cause suffering and premature death to those affected. The risk of suffering from our most common well-being diseases such as cardiovascular disease, type 2 diabetes and neurodegenerative diseases such as Alzheimer's disease increases with overweight and obesity. By far the most effective way to lose weight is by reducing your calorie intake. To maintain a weight loss, behavioural changes, support and a long-term plan are needed. Purpose: The purpose of the study was to investigate whether intermittent calorie restriction (CR) is more effective than continuous CR in terms of weight reduction and maintenance of new weight. Method: A literature study was conducted where the results of four parallel studies were put together. The parallel studies compare the effectiveness of intermittent or continuous CR on weight reduction and maintenance of new weight. Result: In three of the four studies examined, no differences in effectiveness can be demonstrated between intermittent and continuous CR in terms of weight reduction and maintenance of new weight over time. One of the studies found that intermittent CR, in the form of "week on, week off" two weeks at a time, is more effective in weight reduction after 16 weeks of intervention and 6 months later. Conclusion: Like all meta-studies and most studies on the subject, no difference in effectiveness between intermittent and continuous CR can be demonstrated in terms of weight reduction. Nor does one seem superior to the other when it comes to maintaining new weight. More studies over longer time spans with more participants are required to be able to draw conclusions about the type of CR that is most effective in the long term.

Restriction endonucleases are a group of enzymes that cleave DNA at or around specific sequences, which are typically palindromic. A fosmid library was constructed from a metagenome isolated from soil from the Kogelberg Nature Reserve, Western Cape and was functionally screened for restriction endonucleases. Next-generation (NGS) Illumina sequencing technology was used to identify putative endonucleases. The sequence data generated was assembled and analysed using CLC Bio Genomics Workbench and bioinformatics tools (NCBI BLAST, REBASE and MG-RAST). Using these tools, genes encoding restriction-modification systems and endonuclease homologues were discovered. Three genes were identified and were recombinantly produced in Rosetta™ (DE3) pLysS and purified with IMAC using Ni-TED resin and subsequently characterised. These three genes were selected based on the identity percentage when compared to sequences on the NCBI database. Production of Endo8 was scaled up using 2 l fermenter and the purification done using ÄKTA Avant 150 FPLC using a HiScale 50 column packed with Ni-TED resin and the total amount of protein achieved was 58.82 mg.g-1. The productivity achieved at 17 hours (8 h harvest) was 2-fold greater than at 12 hours. Endonuclease activity of endo8 and endo52 was tested, both exhibited strong non-specific activity at 37 °C with an incubation period of 30 min. This work demonstrates that environmental soil samples are a valuable source for discovery of novel enzymes and also the utility of functional metagenomics to discover and purify these enzymes. These endonucleases may contribute to the next generation of reagent enzymes for molecular biology research. / Chemistry / M. Sc. (Life Sciences)

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Diverses études hydrologiques suggèrent que les changements climatiques anticipés entraîneront une diminution des débits d'étiage pour une majorité des rivières du sud du Québec. Cette situation risque de devenir particulièrement préoccupante dans le bassin versant de la rivière Yamaska, car son débit estival est déjà faible. Puisque cette rivière est la principale source d'approvisionnement en eau pour différentes municipalités et les industries qu'elles desservent, des actions doivent être entreprises avant d'en arriver à des situations critiques. Un système d'alerte basé sur des débits seuils jumelé à une prévision météorologique constituerait un outil de gestion puissant. L'élaboration d'un tel système d'alerte nécessite une bonne connaissance de la rivière. Des analyses statistiques des différents épisodes d'étiage survenus dans les années passées permettent d'en faire ressortir les débits historiques ainsi que d'en comprendre le comportement en tarissement. Une revue bibliographique et l'analyse de différents modèles de prévision des débits permettent de déterminer les modèles qui s'appliquent le mieux au bassin de la rivière Yamaska. Des mesures de restriction pour chacun des différents niveaux du système d'alerte proposé doivent être déterminées en s'inspirant des différentes mesures de restriction présentes dans différents pays, tout en tenant compte des facteurs économiques et politiques. Aussi, les critères écologiques et fonctionnels doivent être pris en compte lors de l'établissement de ces débits seuils. Sur la base d'un suivi d'indicateurs et de prévisions associées, ce mémoire propose donc un système d'alerte qui indiquera le moment où des mesures restrictives de prélèvement et de consommation doivent commencer à être appliquées. En plus de constituer un outil puissant pour le bassin versant de la rivière Yamaska, ce prototype de système d'alerte aux étiages pourra être adapté à d'autres bassins versants du Québec enclins aux étiages sévères.

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The APOBEC3 (Apo3) family of proteins are single stranded (ss) DNA cytosine deaminases (C → U). They are grouped into two different structural groups, the single catalytic domain Apo3 enzymes (Apo3A, Apo3C, and Apo3H) and the double catalytic domain Apo3 enzymes (Apo3B, Apo3D, Apo3F, and Apo3G). Apo3G has been implicated in protection from HIV proliferation by becoming encapsidated into budding HIV virions and subsequently mutationally inactivating the synthesized provirus. This largely occurs in the absence of HIV viral infectivity factor (Vif) which mediates the ubiquitination and degradation of Apo3G. Apo3G is a processive enzyme, able to catalyze numerous deaminations in a 5'CCC motif in a single interaction with a substrate. There is a paucity of biochemical data on other Apo3 family members. We performed basic biochemical assays that determined the relative specific activities, processivity, cytosine motif preferences, and binding affinities for DNA, of Apo3A and Apo3H using synthetic DNA substrates in deamination assays. We found Apo3A to be an enzyme with low processivity and Apo3H to be a highly processive enzyme; both of which deaminate a 5'TC motif. Using a reconstituted HIV replication assay we assessed if processivity is needed for efficient restriction of HIV. We were able to demonstrate that each, Apo3G, Apo3A, and Apo3H were able to catalyze deaminations during in vitro reverse transcription. The mutation profile of both Apo3A and Apo3H showed that the 5'TC motif preference was less effective compared to Apo3G in triggering missense and nonsense mutations in the HIV protease active site coding sequence. Nuclear DNA can become deaminated by the related Apo3 family member activation-induced deaminase (AID), when it is present in the nucleus of activated B cells. Apo3A and Apo3H are located in the nucleus but the extent of the damage they cause has only recently been investigated. Here we used an in vitro transcription assay to determine the efficiency of Apo3A and Apo3H deamination during transcription and found that, like AID, they are highly capable of causing deaminations during transcription. Taken together, the results presented here demonstrate that processivity is not necessary for an Apo3 enzyme to catalyze deaminations during HIV reverse transcription and that Apo3A and Apo3H can catalyze deaminations during DNA transcription that could damage host genomic DNA. These results imply a potential cost for maintaining nuclear deaminases.