<p>An important step in tumorgenesis is the acquisition of cellular immortality. Tumor cells accomplish this by activating the enzyme telomerase, and thereby avoiding replicative senescence. The aim of this thesis was to study the activity and regulation of telomerase in a panel of malignant cell types.</p><p>We found that TGF-β1 (transforming growth factor-β1) mediated differential effects on telomerase activity in five ATC (anaplastic thyroid carcinoma) cell lines. Cells that harbored a <i>p53</i> mutation responded by up-regulation of telomerase activity after TGF-β1 treatment, whereas cell lines displaying wt <i>p53 </i>responded by down-regulation of telomerase activity. Thus, these results indicate a possible connection between <i>p53</i> genotype and telomerase response to TGF-β1 treatment. Furthermore, the decreased telomerase activity appeared to be due to transcriptional repression of the <i>hTERT</i> promoter and the increased activity possibly involved hTERT activation via phosphorylation. </p><p>We have previously shown that IFNs (interferons) sensitize MM (multiple myeloma) cells to Fas-mediated apoptosis. In the present investigation both IFN-α and IFN-γ down regulated telomerase activity in the MM cell line U-266-1970. The mechanism underlying the reduction of telomerase activity by IFN was shown to be transcriptional repression of the <i>hTERT </i>gene. We suggest that one potential mechanism whereby IFN sensitize MM cells to Fas-mediated apoptosis is by repressing <i>hTERT</i> activity at the transcriptional level. </p><p>In the next study we demonstrated that basal telomerase activity is not a key determinant of sensitivity to cytotoxic drugs in ESCC (esophageal squamous cell carcinoma) cell lines. Furthermore, we observed no correlation between <i>c-Myc</i> amplification, <i>p53</i> mutations and high telomerase activity levels in these cell lines. </p><p>Finally, neuroblastoma cell lines were shown to up-regulate telomerase activity in response to hypoxic exposure and the main regulatory mechanism was not mediated by increased hTERT mRNA expression. This finding might constitute an adaptive stress response of tumor cells exposed to hypoxia. </p>

An important step in tumorgenesis is the acquisition of cellular immortality. Tumor cells accomplish this by activating the enzyme telomerase, and thereby avoiding replicative senescence. The aim of this thesis was to study the activity and regulation of telomerase in a panel of malignant cell types. We found that TGF-β1 (transforming growth factor-β1) mediated differential effects on telomerase activity in five ATC (anaplastic thyroid carcinoma) cell lines. Cells that harbored a p53 mutation responded by up-regulation of telomerase activity after TGF-β1 treatment, whereas cell lines displaying wt p53 responded by down-regulation of telomerase activity. Thus, these results indicate a possible connection between p53 genotype and telomerase response to TGF-β1 treatment. Furthermore, the decreased telomerase activity appeared to be due to transcriptional repression of the hTERT promoter and the increased activity possibly involved hTERT activation via phosphorylation. We have previously shown that IFNs (interferons) sensitize MM (multiple myeloma) cells to Fas-mediated apoptosis. In the present investigation both IFN-α and IFN-γ down regulated telomerase activity in the MM cell line U-266-1970. The mechanism underlying the reduction of telomerase activity by IFN was shown to be transcriptional repression of the hTERT gene. We suggest that one potential mechanism whereby IFN sensitize MM cells to Fas-mediated apoptosis is by repressing hTERT activity at the transcriptional level. In the next study we demonstrated that basal telomerase activity is not a key determinant of sensitivity to cytotoxic drugs in ESCC (esophageal squamous cell carcinoma) cell lines. Furthermore, we observed no correlation between c-Myc amplification, p53 mutations and high telomerase activity levels in these cell lines. Finally, neuroblastoma cell lines were shown to up-regulate telomerase activity in response to hypoxic exposure and the main regulatory mechanism was not mediated by increased hTERT mRNA expression. This finding might constitute an adaptive stress response of tumor cells exposed to hypoxia.

This dissertation presents a study of the molecular mechanism underlying the highly efficient solar energy conversion processes that occur in the Photosystem I (PS I) reaction centers in plants and bacteria. The primary electron donor P700 is at the heart of solar energy conversion process in PS I and the aim is to obtain a better understanding of the electronic and structural organization of P700 in the ground and excited states. Static Fourier Transform Infra-Red (FTIR) difference spectroscopy (DS) in combination with site directed mutagenesis and Density Functional Theory (DFT) based vibrational frequency simulations were used to investigate how protein interactions such as histidine ligation and hydrogen bonding modulate this organization. (P700+-P700) FTIR DS at 77K were obtained from a series of mutants from the cyanobacterium Synechocystis sp. 6803 (S. 6803) where the amino acid residues near the C=O groups of the two chlorophylls of P700 where specifically changed. (P700+-P700) FTIR DS was also obtained for a set of mutants from C. reinhardtii where the axial ligand to A0-, the primary electron acceptor in PS I was modified. The FTIR DS obtained from these mutants provides information on the axial ligands, the hydrogen bonding status as well as the polarity of the environment of specific functional groups that are part of the chlorophyll molecules that constitute P700. Assignment of the FTIR bands to vibrational modes in specific types of environment is very difficult. In order to assist the assignment of the difference bands in experimental spectra DFT based vibrational mode frequency calculations were undertaken for Chl-a and Chl-a+ model molecular systems under different set of conditions; in the gas phase, in solvents using the Polarizable Continuum Model (PCM), in the presence of explicit solvent molecules using QM/MM methods, and in the presence of axial ligands and hydrogen bonds. DFT methods were also used to calculate the charge, spin and redox properties of Chl-a/Chl-a’ dimer models that are representative of P700, the primary electron donor in PS I.

Ce mémoire présente une méthodologie théorique pour comprendre l'origine de différence de comportement dynamique de complexes alkylidènes, catalyseurs de type Schrock de la métathèse des oléfines, greffés un support de silice amorphe. Dans un travail antérieur, les différences entre les valeurs de l'anisotropie de déplacement chimique (CSA) obtenues par des mesures de RMN du solide et celles estimées par le calcul pour des systèmes figés avaient conduit à suggérer des régimes dynamiques différents pour ces complexes, certains étant proposés comme immobiles, d'autres comme mobiles. Dans le premier groupe se trouve les complexes du molybdène et dans le second les complexes du tungstène, rhénium et tantale. Dans le cadre de cette thèse, nous nous sommes donc attachés à mettre en place une méthodologie pour déterminer ces CSA et donc la nature de la dynamique de chaque système qui conduit au CSA moyenné. Nous nous sommes d'abord intéressés à des systèmes moléculaires pour révéler des interactions non covalentes entre les complexes et le support silice à partir d'une approche de type petit cluster en utilisant divers niveaux de calculs DFT et modèles moléculaires. Cette modélisation moléculaire de la silice étant insuffisante, nous avons entrepris une modélisation de la surface de silice amorphe par dynamique moléculaire classique dont les caractéristiques ont été comparées aux données expérimentales existantes. Le comportement dynamique de ces systèmes greffés sur silice amorphe a été simulé par dynamique moléculaire ab initio QM/MM, couplant une description quantique du complexe organométallique à une description classique du support. Ces études dynamiques ont conduit à des valeurs de CSA moyennées dans le temps de la dynamique. La comparaison de ces valeurs calculées et des valeurs expérimentales a permis d'apporter des éléments de réponse sur l'origine des différences de comportement dynamique de ces complexes alkylidènes. De façon remarquable des mouvements d'ensemble des espèces greffées par rapport à la surface de silice et des modification de la coordination du métal par l'apparition d'interaction agostique contribuent à moyenner le CSA.

L’objectif de ce travail de thèse est le développement en technologie intégrée standard d’une structure de ligne de transmission optimisée en termes de pertes, d’encombrement, de facteur de qualité et surtout du choix du niveau de couplage aux fréquences millimétriques. Cette structure a été nommée CS-CPW (Coupled Slow-wave CoPlanar Waveguide). Dans un premier temps, la théorie ainsi que les modèles électriques des CS-CPW sont présentés. Grâce aux modèles et aux simulations électromagnétiques, des coupleurs directionnels avec plusieurs valeurs de couplage (3 dB, 10 dB, 18 dB) ont été conçus en technologie BiCMOS 55 nm. Ils présentent tous une très bonne directivité, elle est toujours supérieure à 15 dB. Un premier prototype de coupleur a été mesuré à 150 GHz. Dans un deuxième temps, des filtres à la base des lignes couplées ont été développés à 80 GHz en utilisant des lignes CS-CPW. Les résultats des simulations présentent des résultats concurrentiels avec l’état de l’art : 11% de bande passante relative et un facteur non-chargé autour de 25. Finalement, trois projets ont démarré à la base de ces lignes. Ces projets sont actuellement utilisés dans deux travaux de thèse et un stage : un RTPS à 47 GHz, un isolateur à 75 GHz et un balun à 80 GHz. / This work focuses on high-performances CS-CPW (Coupled Slow-wave CoPlanar Waveguide) transmission lines in classical CMOS integrated technologies for the millimiter-wave frequency band. First, the theory as well as the electrical models of the CS-CPW are presented. Thanks to the models and electromagnetic simulations, directional couplers with different coupling levels (3 dB, 10 dB, 18 dB) were designed in BiCMOS 55 nm technology. They have a good directivity, always better than 15 dB. A first prototype of a coupler was measured at 150 GHz presenting good agreement with the simulations. Next, coupled-line base filters were developed at 80 GHz using the CS-CPWs. Simulation present competitive results with the state-of-art: 11% of fractional bandwidth and a unload quality factor of 25. Finally, three projects started based on the CS-CPWs. The projects are currently used in two theses and one internship: a RTPS at 47 GHz, an isolator at 75 GHz and a balun at 80 GHz.

Mr Justice Corbett made a substantial contribution to the South African tax law as he delivered several judgements during his long career on the bench. Starting from the lower ranks as a judge he became Chief Justice of South Africa. Precedents set by his judgements are considered important and indicative of the level of South African tax law. This dissertation observes his background, looks at the operations of the tax court in South Africa and examines whether his judgements were cited and applied in subsequent cases as accepted precedent. International case law is referred to, to compare his judgements with comparable international tax law. / Auditing / M. Comm. (Accounting)

La demande toujours croissante de connectivité et de débit de données requiert une rupture dans la conception des futurs réseaux de communication et systèmes radio. Plusieurs applications émergentes en bande millimétrique, notamment les réseaux mobiles de cinquième génération (5G) et les communications satellites, exigent des antennes large bande qui assurent une grande couverture angulaire, tout en étant à la fois compactes, facilement intégrables et à bas coût.Cette thèse propose des systèmes antennaires multifaisceaux large bande et à très grande couverture angulaire, appelés «Continuous Transverse Stub Antenna» (CTS), pour réaliser un bon compromis de l’ensemble de ces objectifs. L’architecture de l’antenne comprend un réseau de fentes longues excitées par un réseau d’alimentation en chandelier, basé sur des guides d’onde à plans parallèles. Cette structure est excitée par un formateur de faisceaux quasi-optique co-intégré. La première partie du manuscrit présent des nouveaux modèles numériques qui facilitent la conception de chaque sous-système de l’antenne et permettent l’analyse des performances globales, soit en termes d’adaptation, soit en termes de diagrammes de rayonnement. Ces outils sont exploités pour la conception d’antenne et pour étudier les limites en balayage. La thèse se poursuit en présentant de nouvelles solutions technologiques et de nouveaux design pour intégrer les antennes CTS dans des modules multicouches planaires et à faible profil. La conception et la caractérisation de deux antennes intégrées en technologie LTCC pour des points d’accès 5G à 60 GHz sont discutées. L’une des deux est à faisceau fixe, l’autre est à balayage électronique, avec une couverture de ±40°, de faibles lobes secondaires et un niveau élevé de recoupement des faisceaux. Enfin, nous proposons l’association de radomes polarisants planaires à faible profil aux antennes CTS, pour réaliser des systèmes rayonnants en polarisation circulaire. Une méthodologie systématique pour la conception de polarisateurs à très large bande est présentée, ainsi qu’un design couvrant entièrement la bande Ka pour des applications satellites. / The ever-growing demand for fast and seamless connectivity shows the need of new wireless standards and technologies. Novel broadband, wide-angle scanning antennas achieving an optimal trade-off among size, gain, efficiency and costs are crucial to the development of emerging applications at millimeter waves, such as fifth-generation (5G) mobile networks and satellite communications. In this thesis, multibeam parallel-fed continuous transverse stub (CTS) array antennas are proposed as possible candidates for future mm-wave communications and are developed to tackle these requirements. The antenna architecture comprises an array of long slots, a corporate feed network based on parallel plate waveguides (PPWs) and an integrated quasi-optical beamformer. First, novel numerical models for the analysis of each subsystem and of the overall antenna, are presented, which enable an efficient and modular design of CTS antennas. These tools are exploited to derive design guidelines and assess the scanning performance. Then, novel design and technological solutions for the integration of CTS antennas in flat, low-profile multilayer modules are discussed. The design and characterization of two prototypes in LTCC technology, for 60-GHz mobile access points are presented: a fixed beam array and a switched-beam antenna with a field of view of ±40°, low SLLs and high beam overlap. Finally, planar linear-to-circular polarization converters are proposed to realize circularly polarized CTS antenna systems. A procedure to achieve an ultra-wideband, low-loss polarization conversion is outlined and a design for Ka-band satellite application is presented.

Abstract Spontaneous changes in protein systems, such as the binding of a ligand to an enzyme or receptor, are characterized by a decrease of free energy. Despite the recent developments in computing power and methodology, it remains challenging to accurately estimate free energy changes. Major issues are still concerned with the accuracy of the underlying model to describe the protein system and how well the calculation in fact emulates the behaviour of the system. This thesis is largely concerned with the quality of current free energy calculation methods as applied to protein-ligand systems. Several methodologies were employed to calculate Gibbs standard free energies of binding for a collection of protein-ligand complexes, for which experimental affinities were available. Calculations were performed using system description with different levels of accuracy and included a continuum approach, which considers the protein and the ligand at the atomic level but includes solvent as a polarizable continuum, and an all-atom approach that relies on molecular dynamics simulations. In most such applications, the effects of ionic strength are neglected. However, the severity of this approximation, in particular when calculating free energies of charged ligands, is not very clear. The issue of incorporating ionic strength in free energy calculations by means of explicit ions was investigated in greater detail and considerable attention was given to the affinities of charged peptides in the presence of explicit counter-ions. A second common approximation is concerned with the description of ligands that exhibit multiple protonation states. Because most of current methods do not model changes in the acid dissociation constants of titrating groups upon binding, protonation equilibria of such ligands are not taken into account in free energy calculations. The implications of this approximation when predicting affinities were analysed. Finally, when calculating free energies of binding, a correct description of the interactions between the protein and the ligand is of fundamental importance. However, active sites of enzymes, where strained conformations may hold a functional role, are not always accurately modelled by molecular mechanics force fields. The case of a strained planar proline in the active site of triosephosphate isomerase was investigated using an hybrid quantum mechanics/molecular mechanics method, which implies a higher level of accuracy.

Cílem této práce je popsat současný stav v oblasti mikrovlnných komponent na bázi textilu a poté také autorův příspěvek k výzkumu. V tomto studijním oboru je třeba řešit mnoho problémů, a proto jsou stanoveny velmi konkrétní cíle, které by měly tvořit kompletní komunikační systém, který by byl snadno integrovatelný do čalounění uvnitř letadla nebo jiného vozidla. První kapitola je zaměřena na použití relativně nízkofrekvenčních obvodů v pásmu UHF a vysokofrekvenční sklízení energie. Po simulacích následují praktická měření. Další kapitola charakterizuje nové 3D pletené textilie a jejich vysokofrekveční vlastnosti a modelování na velmi vysokých frekvencích, které jsou žádoucí pro užití jako komunikační kanály. S prakticky ověřeným a numericky popsaným textilním substrátem pro použití v pásmu SHF je představena nový vlnovod integrovaný do textilu na bázi tisknutelného umělého magnetického vodiče s praktickou metodikou návrhu. Po ověření byl vlnovod použit pro základní dělič výkonu, dvě různé antény a dva typy senzorů. Všechny návrhy byly vyrobeny a testovány s uspokojivými výsledky.

For three decades, the ongoing HIV pandemic has taken the lives of tens of millions of people. Still, more tens of millions are fighting this incurable disease today. Current failures in combating this global problem are caused mainly by the virus's extreme ability of mutation, its very effective molecular shield which repels the immune system's attacks, and its immense variability. A breakthrough, achieved relatively recently, is the discovery of the so-called broadly neutralizing antibodies against HIV-1, which carry a very efficient and broad neutralizing response. So far, it's not known how to elucidate the production of these antibodies in the infected hosts to quell or altogether eliminate the virus. This work deals with experimental results, which led to both in vivo and in vitro proof-of-concept of the so-called protein mimetics, the ability to imitate viral surface epitopes, and therefore stimulate an efficient immune response carried by targeted broadly neutralizing antibodies. This effect is mediated by recombinant binding proteins, based on the Myomedin scaffold. This work describes the selection and characterization of these binding proteins mimicking the epitopes of one of the most effective broadly neutralizing antibodies, 10E8. It shows that the binding affinities of selected...