No description available.

No description available.

The motivation of this study lies in the necessity for a microfocal therapy to specifically target diseased areas in vascular pathologies such as age-related macular degeneration (AMD). AMD is the most common cause of legal blindness among people over the age of 60 in developed countries. This degenerative condition affects the macula, the central region of the retina, severely impairing detailed vision and hindering everyday activities. Worldwide, 25-30 million people live with some form of AMD. Among them, ~10% suffer from the more advanced and damaging form, wet-AMD, which causes rapid and severe loss of central vision. To date, there is no cure or long-term alternative for this degenerative disease despite intensive research efforts. With recent developments in biophysical tools and experimental procedures, in this study, we demonstrate a highly-localized therapeutic option: two-photon (2-photon) photodynamic therapy (PDT) that could be advantageous for the cure of wet-AMD, either alone or in combination with recently discovered anti-angiogenic therapies. This new approach offers selective targeting of the diseased area, thus minimizing damage to the surrounding sensitive healthy eye tissues, which is a major concern with the clinically-used, standard wide-beam, one-photon (1-photon) PDT. The objective of the research was to test the feasibility of microfocal 1-photon and the inherently localized 2-photon PDT, their optimization and also to evaluate the efficacy of existing 1-photon and novel 2-photon photosensitizers. In this thesis, I illustrated the in vitro (endothelial cell monolayer) and in vivo (window chamber mouse (WCM)) models that can be used to quantitatively compare the 2-photon efficiency of photosensitizers. Using the in vitro model, I compared the 2-photon efficacy of clinically used 1-photon PDT drugs Photofrin and Visudyne, and showed that the Visudyne is an order of magnitude better 2-photon photosensitizer than Photofrin. With the WCM model, I demonstrated a novel designer 2-photon photosensitizer is 20 times more efficient than Visudyne for single vessel occlusion. I also generated the drug and light dose reciprocity curve for localized single-vessel microfocal PDT. This is a necessary step towards applying the method to the relevant ocular models of AMD, which is the next phase for this research.

De part, leur implication dans la prolifération cellulaire, l'invasion et leur surexpression dans de nombreux cancers, les récepteurs à tyrosine kinase de la famille HER constituent des cibles de choix en oncologie. Parmi ces récepteurs, le récepteur HER3 semble pertinent car il est impliqué dans la tumorigenèse de nombreux cancers (sein, ovaire, pancréas, mélanome…) et il est associé à un mauvais pronostic. De plus, la surexpression du récepteur HER3 est souvent associée à l'apparition de résistance aux thérapies ciblées. Nous avons sélectionné plusieurs anticorps anti-HER3 humains et murins respectivement par « phage display » et fusion cellulaire. Ces derniers reconnaissent spécifiqument le récepteur HER3. Parmi les nombreux anticorps découvert, nous avons sélectionné un anticorps anti-HER3 humain H4B-121 et 2 anticorps anti-HER3 murins 9F7-F11 et 16D3-C1. Ces derniers ont la capacité à faire régresser des tumeurs épidermoide, pancréatique et triple négative chez la souris, en présence ou en abscence de neuréguline et indépendamment du status HER2 et P53/PTEN. Cette inhibition est possible grâce à un blocage du cycle cellulaire en phase G1, une inhibition de la prolifération ainsi qu'une induction de l'apoptose. Ces trois anticorps sont capables de bloquer l'hétérodimérisation des récepteurs HER2/HER3 ainsi que la phosphorylation du récepteur HER3. Ils sont aussi capables d'inhiber la phosphorylation de la protéine AKT ainsi que de ses cibles (MDM2, FOXO et XIAP). De plus, nous avons montré que nos anticorps sont capables de lyser les cellules tumorales par ADCC (Antibody-dependent cell-mediated cytotoxicity). Cette étude démontre que ces anticorps anti-HER3 représentent une nouvelle thérapie pour les cancers du pancréas et du sein triple négatif. / Due to their implication in the cellular proliferation, the invasion and their surexpression in numerous cancers, the tyrosine kinase receptors of HER family constitute one of the best targets in oncology. Within this family, the human epidermal growth factor receptor 3 (HER3) plays a role in tumorigenesis of different cancers (Breast, melanoma, pancreas and ovary). This receptor is implicated in drug resistance and he is over expressed in cancers that are not eligible for the currently approved targeted therapies. To this end, we generated specific antibodies (Abs) against domain 1 (D1) and domain 3 (D3) of HER3 that recognize epitopes that do not overlap with the neuregulin-binding site. The fully human H4B-121 Ab and the mouse monoclonal Abs 16D3-C1 and 9F7-F11 inhibited tumor growth in nude mice xenografted with epidermoid, pancreatic, or triple-negative breast cancer cells independently of NRG addiction, HER2 status and p53/PTEN mutations. The combination of one anti-HER3 Ab and trastuzumab improved tumor growth inhibition in mice xenografted with HER2(low) cancer cell lines, for which trastuzumab alone shows no or moderate efficiency. Ab-induced disruption of tumor growth was associated with G1 cell cycle arrest, proliferation inhibition, and apoptosis of cancer cells. Anti-HER3 Abs blocked HER2/HER3 heterodimerization and HER3 phosphorylation at the cell membrane, leading to inhibition of phosphorylation of the downstream AKT targets murine double minute 2, X-linked inhibitor of apoptosis, and forkhead box O1. Anti-HER3 Abs can also induice antibody dependant cell-mediated cytotoxicity. This study demonstrates that anti-HER3 D1 and D3 Abs could represent a new option for immunotherapy of pancreatic and triple-negative breast cancers.

Research report (M.Com. (Development Theory and Policy))--University of the Witwatersrand, Faculty of Commerce, Law and Management, School of Business and Economic Sciences, 2013. / The global financial crisis has shaken not only the foundations of the financial system but also elements of macroeconomic stability, particularly monetary policy as it relates to the central bank institution and its fundamental operations. This paper is centred on examining the aspects of the inflation-targeting framework both theoretically and practically, as the fulcrum around which modern central banking functions, in the context of the crisis and the economic conditions thereafter. This discussion is based on the idea that there exist spaces for broadening and extending the mandate of the central bank beyond inflation targeting and that crisis conditions have gone to show that.

The cannabinoid system is involved in many functions of mammalian brain, such as learning and memory, pain perception and 'locomotion. The "brain type" cannabinoid receptor CB 1 is one of the key elements of the cannabinoid system. In this Thesis, some aspects of the neurobiology of mouse CB 1 are described. CB 1 mRNA distribution was analysed by single and double in situ hybridization (ISH), revealing the expression of the receptor in specific neuronal subpopulations. This expression pattern suggests many putative functional cross-talks between the cannabinoid system and other signalling molecules in the brain, such as glutamate, GABA, cholecystokinin and nitric oxide (NO). The putative functional interactions of the cannabinoid system with the NO pathway was studied by pharmacological treatment of neuronal NO synthase (nNOS) mutant mice with the CBI agonist A9-tetrahydrocannabinol (A9-THC). The results showed that nNOS is necessary for some central effects of A9-THC. Moreover, ISH analysis revealed. that nNOS-deficient mice contain levels of CBI lower than normal in selected brain regions. A "conditional" targeting approach was developed to gain insights into the specific functions of CB 1 in mouse brain. By gene targeting experiments, two mutant lines were obtained. The "Flox CB 1" mouse line, containing the whole open reading frame of CB I flanked by two loxP sites will be the key tool for the generation of mouse mutants with a spatiotemporal-restricted deletion of CB I. The "CBN" mice, carrying a "null" mutation of CB 1, were used for a study aimed to clarify some aspects of the in vitro neuroprotective activity of cannabinoids and, in particular, the involvement of CB 1. In vitro oxidative stress assays were performed on cell lines and on primary neuronal cultures derived from homozygous CBN/CBN mice and wild type littermates. The results indicate a differential protective activity of cannabinoids on cell lines and primary cultures. However, CBI does not appear to be involved in the in vitro leuroprotective effects of cannabinoids.

"The use of behavioral targeting practices provides ad networks with the opportunity to tailor ads to the individual characteristics of users. As privacy concerns over behavioral targeting have been growing lately, an increasing number of ad networks offer ad preferences managers (APMs) that show collected and/or inferred information about users. The focus of our study is to investigate the accuracy and completeness of the information contained in such APMs. On the basis of our experimental results, we propose a structured methodology for APM validation. We also assess how third parties render ads based on users’ browsing behavior. Our findings reveal cases in which even sensitive information is leaked as part of an HTTP header and is used to serve ads on multiple sites. The third parties examined in this study include an intent-focused data exchange (BlueKai) and a social network (Facebook) along with the ad networks owned by AOL, Google, and Yahoo!."

Diplomová práce ?Marketingová strategie Renaultu: výzvy střednědobého plánu? si klade za cíl analyzovat marketingovou strategii společnosti Renault. Práce nejdříve popisuje společnost Renault jako takovou. Zaměřuje se na historii společnosti, firemní strategii a aktuální střednědobé cíle. Následující část popisuje přístup firmy Renault k výzkumu trhu. Zde je popsán jak kontinuální, tak jednorázový výzkum. Poté práce detailně rozebírá segmentaci, targeting a positioning. Všechny tři strategické marketingové přístupy jsou analyzovány teoreticky i prakticky, tedy z pohledu firmy Renault.

<p>As a consequence of the presence of both mitochondria and chloroplasts in plant cells there is a higher sorting requirement in a plant cell than that in a non-plant cell. Reflecting this, protein import to mitochondria and chloroplasts has been shown to be highly specific. However, there is a group of proteins which are encoded by a single gene in the nucleus, translated in the cytosol and targeted to both mitochondria and chloroplasts. These proteins are referred to as dual targeted proteins. The first protein shown to be dual targeted was pea glutathione reductase (GR). The focus of this thesis is the targeting properties of the dual targeted protein glutathione reductase.</p><p>In order to overcome the limitations with traditional in vitro import systems we have developed an import system for simultaneous import of precursor proteins into mitochondria and chloroplasts (dual import system). The chloroplastic precursor of the small subunit of ribulose bisphosphate carboxylase/oxygenase (SSU) was mis-targeted to pea mitochondria in a single import system, but was imported only into chloroplasts in the dual system. The dual GR reductase precursor was targeted to both mitochondria and chloroplasts in both the single and dual import system.</p><p>We have investigated the targeting and processing properties of the GR targeting signal. Using N-terminal truncations we have demonstrated that the GR targeting signal has a domain organisation. Our results show that GR has evolved a dual targeting signal with the C-terminal part being sufficient for chloroplast import, the internal part required for the mitochondrial import and the N-terminal part housing a “fine-tuning” function. Furthermore, we have constructed a range of point mutations on the GR signal sequence changing positive amino acid residues and stretches of hydrophobic amino acid residues. Overall single mutations had a greater effect on mitochondrial import compared to import into chloroplasts. We have also shown that the recognition of the GR processing site differs between MPP and SPP. Single amino acid substitutions in the vicinity of the processing site clearly affected processing by MPP while processing by SPP showed low sensitivity to single mutations.</p>

As a consequence of the presence of both mitochondria and chloroplasts in plant cells there is a higher sorting requirement in a plant cell than that in a non-plant cell. Reflecting this, protein import to mitochondria and chloroplasts has been shown to be highly specific. However, there is a group of proteins which are encoded by a single gene in the nucleus, translated in the cytosol and targeted to both mitochondria and chloroplasts. These proteins are referred to as dual targeted proteins. The first protein shown to be dual targeted was pea glutathione reductase (GR). The focus of this thesis is the targeting properties of the dual targeted protein glutathione reductase. In order to overcome the limitations with traditional in vitro import systems we have developed an import system for simultaneous import of precursor proteins into mitochondria and chloroplasts (dual import system). The chloroplastic precursor of the small subunit of ribulose bisphosphate carboxylase/oxygenase (SSU) was mis-targeted to pea mitochondria in a single import system, but was imported only into chloroplasts in the dual system. The dual GR reductase precursor was targeted to both mitochondria and chloroplasts in both the single and dual import system. We have investigated the targeting and processing properties of the GR targeting signal. Using N-terminal truncations we have demonstrated that the GR targeting signal has a domain organisation. Our results show that GR has evolved a dual targeting signal with the C-terminal part being sufficient for chloroplast import, the internal part required for the mitochondrial import and the N-terminal part housing a “fine-tuning” function. Furthermore, we have constructed a range of point mutations on the GR signal sequence changing positive amino acid residues and stretches of hydrophobic amino acid residues. Overall single mutations had a greater effect on mitochondrial import compared to import into chloroplasts. We have also shown that the recognition of the GR processing site differs between MPP and SPP. Single amino acid substitutions in the vicinity of the processing site clearly affected processing by MPP while processing by SPP showed low sensitivity to single mutations.