L’étude des mécanismes aboutissant à la tumorigénèse a permis de révéler, dans beaucoup de cancers, une amplification ou une mutation de divers oncogènes, avec pour conséquence des capacités de prolifération et de survie accrues pour la cellule tumorale. L’identification des protéines kinases comme étant des éléments centraux de ces processus en ont fait des cibles thérapeutiques prometteuses. Plusieurs inhibiteurs ciblant de façon plus ou moins spécifique les tyrosines kinases oncogéniques ont ainsi été développés. Parmi eux, l’imatinib mesylate (Gleevec®, Novartis) a constitué la première chimiothérapie ciblée. Il correspond aujourd’hui au traitement de première intention contre la LMC. Cependant, malgré sa très grande efficacité, il est apparu que certains mécanismes de résistances pouvaient être mis en place pour diminuer son effet pro-apoptotique. Le travail de cette thèse a consisté à mieux comprendre les mécanismes d’apoptose induits par les inhibiteurs de tyrosine kinases (ITK), et à rechercher quelles voies alternatives de survie devront être bloquées pour leur assurer une meilleure efficacité. Trois modèles ont été utilisés : la leucémie myéloïde chronique (LMC), les leucémies aiguës myéloïdes (LAM) et les glioblastomes (GBM). La LMC a été utilisé comme modèle et la démarche utilisé pour essayer d’augmenter l’efficacité des ITK, a été transposée aux modèles des LAM et des GBM. L’ensemble des résultats obtenus a démontré qu’une meilleure compréhension de la réponse apoptotique et des mécanismes de résistance permettait l’identification de nouvelles cibles thérapeutiques. Nous avons pu observer que, tout en favorisant la diminution des doses de molécules administrées, l’inhibition simultanée de plusieurs cibles apportait plusieurs bénéfices. Elle permet d’augmenter l’action pro-apoptotique des ITK, de contrer certains mécanismes de résistances, d’atteindre la cellule souche cancéreuse résistance et par conséquent de cibler simultanément des populations a plusieurs de stades de différenciation. / Protein kinases have been identified as playing fundamental roles in cancer development, suggesting that they could represent a promising therapeutic target. Several kinase inhibitors have been developed and the most successful of them, by far, is Gleevec® (imatinib, STI57; Novartis), a BCR-ABL inhibitor. It is currently used as the treatment of reference for chronic myeloid leukemia. However, despite a huge efficiency, some resistance mechanisms could be used to decrease its pro-apopototic effect. The global aim of my PhD was to understand the apoptotic mechanisms induced by tyrosine kinase inhibitors (TKI) to identify new potential therapeutic targets. I work on three different tumors: Chronic Myeloid Leukemia (CML), Acute Myeloid Leukemia (AML) and Glioblastomas (GBM). CML has been used as a model and the approach followed to increase TKI efficiency has been transposed to AML and GBM models. Altogether, our results showed that a better understanding of apoptotic response and resistance mechanisms could lead to the identification of new therapeutic targets. We observed that combination therapy brings several benefits. It allows to increase the TKI-induced apoptotic response, to counter some resistance mechanism, to reach the resistant cancer stem cells, and thus, to target simultaneously several populations in the tumour.

Le RAFT-RGD radiomarqué avec un émetteur β- comme nouvel agent de radiothérapie interne vectorisée. L'intégrine αvβ3 est fortement impliquée en oncogenèse à travers son rôle dans la néoangiogenèse tumorale, dans la prolifération et la survie des cellules cancéreuses et dans le processus métastatique. L'intégrine αvβ3 est exprimée faiblement dans la plupart des tissus. Par contre, elle est fortement exprimée par les cellules endothéliales activées lors de l'angiogenèse et par les cellules de nombreux types de cancers invasifs. Ces caractéristiques font de l'intégrine αvβ3 une excellente cible pour l'imagerie et la thérapie de ces tumeurs. Le RAFT-RGD (Regioselectively Addressable Functionalized Template-(cyclo-[RGDfK])4) est un derivé polypeptidique constitué de quatre peptides cyclo-RGD (spécifiques de l'intégrine αvβ3) fixés sur un groupe porteur RAFT. Le RAFT-RGD cible spécifiquement l'intégrine αvβ3 in vitro et in vivo et permet la détection par imagerie nucléaire ou par fluorescence de tumeurs exprimant αvβ3 sur des modèles précliniques. Le RAFT-RGD un excellent vecteur potentiel pour cibler les tumeurs exprimant αvβ3 et pour y délivrer des traitements, que ce soit des molécules de chimiothérapie ou des radionucléides de thérapie. Cette étude est la première à évaluer le potentiel thérapeutique du RAFT-RGD radiomarqué avec un émetteur β- sur un modèle de souris Nude porteuses de tumeurs exprimant l'intégrine αvβ3. Une injection de 37 MBq de 90Y-RAFT-RGD ou de 177Lu-RAFT-RGD permet de ralentir significativement la croissance de tumeurs exprimant l'intégrine αvβ3 par rapport aux souris contrôles non traitées ou traitées avec la même activité de la molécule de contrôle non spécifique de la cible, le RAFT-RAD. En comparaison, une injection de 30 MBq de 90Y-RAFT-RGD ne permet pas de ralentir la croissance de tumeurs n'exprimant pas l'intégrine αvβ3. Le RAFT-RGD présente un bon potentiel pour le traitement de tumeurs exprimant l'intégrine αvβ3 lorsqu'il est radiomarqué avec des émetteurs β-. Mots clés : intégrine αvβ3, RAFT-RGD, ciblage tumoral, radiothérapie interne vectorisée. / Β- emitters radiolabeled RAFT-RGD as new agents for internal targeted radiotherapy. The αvβ3 integrin is known to play an important role in tumor-induced angiogenesis, tumor proliferation, survival and metastasis. Because of its overexpression on neoendothelial cells such as those present in growing tumors, as well as on tumor cells of various origins, αvβ3 integrin is an attractive molecular target for diagnosis and therapy of the rapidly growing and metastatic tumors. A tetrameric RGD-based peptide, regioselectively addressable functionalized template-(cyclo-[RGDfK])4 (RAFT-RGD), specifically targets integrin αvβ3 in vitro and in vivo. RAFT-RGD has been used for tumor imaging and drug targeting. This study is the first to evaluate the therapeutic potential of the β- emitters radiolabeled tetrameric RGD peptide RAFT-RGD in a Nude mouse model of αvβ3-expressing tumors. An injection of 37 MBq of 90Y-RAFT-RGD or 177Lu-RAFT-RGD in mice with αvβ3-positive tumors caused a significant growth delay as compared with mice treated with 37 MBq of 90Y-RAFT-RAD or 177Lu-RAFT-RAD or untreated mice. In comparison, an injection of 30 MBq of 90Y-RAFT-RGD had no efficacy for the treatment of αvβ3-negative tumors. 90Y-RAFT-RGD and 177Lu-RAFT-RGD are potent αvβ3-expressing tumor targeting agents for internal targeted radiotherapy. Keys words : integrin αvβ3, RAFT-RGD, tumour targeting, internal targeted radiotherapy.

Depuis leur introduction, les agents de contraste ont révolutionné l'imagerie échographique. Ils sont composés de microbulles gazeuses, qui injectés par voie intraveineuse dans le sang, ils améliorent l'image échographique. Une autre application pour laquelle les caractéristiques physiques des agents de contraste sont exploitées est l'imagerie ciblée. Une approche basée sur l'utilisation de ligands intégrés à la paroi des microbulles, celles-ci adhérent aux facteurs de surfaces moléculaires surexprimés par les cellules endothéliales qui tapissent la paroi interne des vaisseaux sanguins. Pour pouvoir distinguer ces microbulles de celles qui circulent librement, elles doivent réfléchir un signal acoustique suffisamment intense. Cependant, le faible taux d'adhérence des microbulles engendre une réduction du signal acoustique. Pour résoudre ce problème, il est important de déterminer l'effet des parois sur leurs dynamiques acoustiques. Dans cette thèse, nous avons étudié l’effet des parois élastiques sur le comportement dynamique des microbulles constituant les agents de contraste. Dans un premier temps, un modèle théorique représentant une paroi avec une épaisseur finie a été développé. Il a été démontré que l’amplitude de l’écho rétrodiffusé par une microbulle proche d’une paroi avec une épaisseur finie est inférieure à celui d’une microbulle se trouvant dans un fluide infini. D'autres parts, pour représenter la paroi d’un vaisseau sanguin, les propriétés mécaniques de la paroi élastique ont été intégrées au modèle. Il a été observé que la fréquence de résonance d’une microbulle proche d’une paroi est supérieure à celle dans un fluide infini. Par la suite, nous avons étudié l’effet de trois types de parois sur le comportement d’une microbulle parmi lesquelles la paroi d'OptiCell communément utilisée en expérimentations ultrasonores. Les résultats ont montré que la microbulle proche de la paroi d’OptiCell diffuse un écho supérieur à celui de la microbulle éloignée de la paroi, lorsque la fréquence d’excitation est au-dessus de sa fréquence de résonance. Nous avons constaté aussi que les petites bulles sont plus sensibles à la proximité de la paroi. Par la suite, nous avons développé un modèle décrivant une microbulle attachée à une paroi élastique. Nous avons montré que le contact direct de la bulle avec la paroi induit une diminution de l'écho par rapport à la même bulle dans un liquide infini. Le contact direct de la bulle avec la paroi engendre une augmentation de la fréquence de résonance part rapport à une bulle sans contact direct. Enfin, une étude expérimentale a montré l'avantage de l'imagerie sous-harmonique pour différencier les microbulles attachées des microbulles libres. / Since they were introducted, contrast agents have revolutionized the ultrasound imaging. They are composed of tiny gaseous microbubbles and when injected intravenously into the blood, they improve the ultrasound image. Targeted imaging is another application based on the physical characteristics of contrast agents. This approach is based on the ligands incorporation into the microbubbles shell. The microbubble attach to the molecular factors overexpressed by endothelial cells, covering the inner wall of blood vessels. To distinguish these microbubbles from those freely circulating, attached microbubble have to produce an acoustic signal that is sufficiently strong. However, the low microbubbles adhesion induces a decrease of the acoustic signal. To make it possible, it is important to determine the effect of the elastic wall on their acoustic response. This thesis aimed to study the effect of elastic walls on the ultrasonic behavior of targeted microbubbles. First, a theoretical model describing a wall with finite thickness was developed. It has been shown that the scattered echo amplitude by a microbubble near a wall with finite thickness is small in comparison to the echo from a microbubble located in an infinite fluid. Furthermore, and in order to account for the effect of blood vessel wall, the mechanical properties of the wall have been incorporated into the model. The results showed that the resonane frequency of a microbubble near the wall is higher than the resonanace of the same microbubble in an infinite medium. Subsequently, we studied the effect of three types of walls on the microbubble behavior including the wall of OptiCell chamber which is commonly used in ultrasonic experiments. We have shown that microbubbles near the OptiCell wall diffuses a higher echo than those far from the wall when the excitation frequency is above the microbubble resonance frequency. On the other side, we observed that small microbubbles to the presence of the wall. Afterward, we developed a model describing a microbubble attached to the wall. We have shown that the microbubble in direct contact with the wall induces a decrease of the echo amplitude compared to the same bubble in infinite liquid. Moreover, the direct contact of the bubble with the wall generates an increase of the resonance frequency relative to a bubble without direct contact. Finally, an experimental study has shown the advantage of the subharmonic imaging to differentiate attached microbubbles from the free ones.

Within the fight against terrorism, conflicts may arise between the interest of the state to maximize the effectiveness of methods of combating terrorism and the international standard of human rights. One of the methods that some states carry out to combat terrorism is targeted killing. Legitimate effort of states to suppress terrorism and may not lead to violations of human rights. The thesis aims to answer the question: "Is targeted killing a permissible method of combating terrorism, in terms of the lex lata of human rights law, humanitarian law and rules for the use of force between states?" Regarding the methodology of the thesis I was inspired by the books: "Vědecká propedeutika pro právníky" by Viktor Knapp," Metodologie vědy" by František Ochrana and "Jak studovat politiku" by Peter Drulák. The thesis consists of four chapters. The objective of the first chapter is descriptive. The first chapter should clarify the terminology and content of key concepts. The content of the second chapter is an analysis of the legality of targeted killings under human rights law. The third chapter examines the conditions of legality of targeted killings in humanitarian law. Chapter Four deals with the admissibility of targeted killings in the light of law of interstate force. Through an analysis of...

Nous nous proposons, dans le cadre de cette thèse d'examiner des modèles économiques dans lesquels les firmes ont recours à différentes approches de publicité. Nous analysons les effets des investissements et des tarifs publicitaires sur les stratégies des annonceurs et des médias et nous étudions les différentes configurations de marché qui émergent à l'équilibre. Nous nous focalisons dans les Chapitres 2, 3 et 4 sur la publicité informative ciblée. Dans les chapitres suivants nous considérons que la publicité est persuasive et qu'elle peut créer des externalités aussi bien positives que négatives.Ces externalités peuvent être exogènes comme c'est le cas dans le Chapitre 5 ou endogènes (Chapitre 6).Plus précisément, nous examinons, dans le Chapitre 2, dans le cadre d'une différenciation horizontale, la concurrence en prix et en publicité informative ciblée. Dans le Chapitre 3, nous caractérisons l'équilibre d'un média qui a le choix entre éditer un seul journal général publié dans deux localisations géographiques ou bien éditer un journal local dans chaque localisation, donnant ainsi la possibilité aux annonceurs d'acheter des espaces publicitaires séparés dans chaque édition ou un package publicitaire pour les deux éditions. Nous posons dans le Chapitre 4 le problème du signal de la qualité d'une firme qui lance un nouveau produit, dont la qualité n'est pas connue par les consommateurs potentiels. La firme signale sa qualité à travers une publicité informative ciblée en choisissant la taille du marché niche. L'objectif du Chapitre 5 est de mettre en évidence les interactions directes qui existent entre le marché des médias et le marché des producteurs en considérant que le marché est caractérisé par des externalités de publicité aussi bien positives que négatives. Dans le dernier chapitre, nous étudions les incitations de deux firmes, différenciées par la qualité, à investir dans deux types de publicité : une publicité comparative et une publicité non-comparative. / In this thesis, we develop economic models in which firms invest in different types of advertising. We analyze the effects of the advertising investments and prices on the advertisers and media strategies. In addition, we study the various market configurations emerging at equilibrium. On this basis, we focus, in Chapters 2, 3 and 4, on the targeted informative advertising. In the following chapters, we consider persuasive advertising that create negative and positive externalities. These externalities may be exogenous (Chapter 5) or endogenous (Chapter 6).More precisely, we investigate in Chapter 2, through a horizontal differentiation framework, the price competition and targeted advertising investments when firms are able to perfectly target each type of consumer. In Chapter 3, we model a situation in which a printed media has the choice between editing a single national newspaper published in two cities, or editing two local newspapers allowing thus advertisers to buy separated or bundled ads. In Chapter 4, we investigate the best signalling strategy of a monopoly when introducing a new product with unobservable quality. The firm signals its quality to the potential consumers through informative targeted advertising i.e. by choosing the size of the reached consumer's market. The aim of chapter 5 is to highlight the strategic interaction between media and product markets when the market exhibits positive and negative advertising externalities. The last Chapter is interested in the optimal persuasive advertising and pricing decisions of two vertically differentiated firms given that each firm has the choice to advertise in comparative and/or non-comparative advertising.

The EU has for some time imposed sanctions against individiduals, i.e. targeted sanctions. These sanctions have had impacts on the listed individual in forms of limited procedural rights due to inter alia the limited disclosure of evidence. Improvements in the field were introduced by Kadi. Yet it remains difficult for the individual who wants to challenge a listing. The purpose of this essay is to examine how to obtain mechanisms for an effective judicial review in targeted sanctions cases when there is a non-disclosure of evidence to the EU Courts by the EU institutions. This essay will also discuss how the Courts standard of review could look like after Kadi. It will also address the need for an increased responsibility of the EU institutions, in the matter concerning the access to confidential information. This relates to questions regarding the transparency in the Union. This essay suggests that the mechanisms needed to acquire a higher intensity of review as well as an effective judicial review could be the use of closed material procedures and special advocates.

A research report submitted to the Wits School of Education, Faculty of Humanities, University of the Witwatersrand, in partial fulfilment of the requirements for the degree of Master of Education by combination of coursework and research. Johannesburg 2013 / This study investigates and addresses the issue of bullying of teachers by learners, which is also referred to as teacher-targeted bullying (TTB). The purpose of this study is to obtain data relating to the nature and extent of teacher-targeted bullying in a High School in Gauteng West. The setting is a former Model C, co-educational school, consisting of 1 354 learners (Grade 8 to Grade 12). The ethnicity of the School is predominantly Black, with 60% Black learners to 40% Caucasian, and the gender ratio of learners in the school is 706 girls to 648 boys. The current teacher population consists of a staff quotient of seventy-five teachers (75), twenty (20) male and fifty-five (55) female teachers, who are employed at the school, which will also be known as “the workplace” for the purposes of this research. The procedure for this research entailed using a teacher group of seventy (70) teachers, of whom fifty four (54) volunteered to complete the questionnaire, allowing an in-depth response by establishing factors contributing to the difficulties of learner bullying experienced by teachers; the effect on teacher performance, morale and student learning; and the support needed from education management for teachers experiencing bullying by their learners. From these questionnaires, the process was extended to a semi-structured interview, in order to follow up on the questionnaire. This allowed an understanding of the teacher’s life experiences or situations as experienced in their own words, for the twelve (12) volunteer teachers who had experienced bullying by learners. In this mixed method design, quantitative results enhanced generalizability, and qualitative results helped to explain how teachers experienced teacher-targeted bullying by learners in this Gauteng West High School. The research findings indicated that, prominent in this study, is the stress reported by teachers relating to the disciplining of learners. The level of reported learner-on-teacher-bullying was more evident in the classroom. It was also reported that all Grades were problematic for teachers, but the level of reported stress was more evident with Grade 9 male learners in same-gender classes. Furthermore, difficulties within this school environment, as well as the effect on teachers’ performance, morale and support from school management were established. In an effort to respond to the epidemic of learners who target and bully teachers, a reconceptualisation of the learner-on-teacher-bullying construct in the School under review, as well as other schools in South Africa, is needed. Thus bullying of teachers by learners must be tackled as a whole-school issue, with the focus on positive learner behaviour, good support structures, and active leadership, ensuring that staff are not only supported, but are also seen to be supported, by the various role players.

Le cancer pancréatique représente la cinquième cause de décès par cancer dans les pays occidentaux. Son mauvais pronostic (survie à 5 ans inférieure à 3,5 % des cas) est dû à l’absence de facteurs de risques spécifiques interdisant une prévention efficace, et à un diagnostic tardif qui révèle un cancer agressif chez environ 90% des patients. Actuellement, le seul traitement curatif de ce cancer est la chirurgie, mais celle-ci ne peut être envisagée que dans 10 à 15 % des cas. L’adressage de molécules thérapeutiques vers l’organe, le tissu ou la cellule malade constitue aujourd’hui un défi majeur pour le traitement des maladies humaines notamment infectieuses, cancéreuses ou d’origine génétique. C’est pour ces raisons que le développement de nanotechnologies, en tant que vecteurs de médicaments, a pris un essor considérable au cours des dernières années. Dans ce contexte, le concept de squalènisation repose sur le couplage chimique entre le squalène (SQ), un lipide naturel précurseur de la synthèse du cholestérol, et des principes actifs (notamment des molécules anticancéreuses). Les bioconjugués ainsi formés sont alors capables de s’auto-assembler en solution aqueuse pour former des nanoparticules stables de diamètres compris entre 100 et 300 nm. L’exemple de référence dans ce domaine est la nanoparticule de gemcitabine-squalène (SQdFdC) qui a donné lieu à des résultats spectaculaires in vitro sur des lignées de cellules cancéreuses humaines In vivo, les nanoparticules de gemcitabine-squalène se sont avérées beaucoup plus efficaces que la gemcitabine libre sur des tumeurs solides greffées par voie sous-cutanée ainsi que sur des modèles murins de leucémies agressives métastatiques.Au vu de ces résultats encourageants, le projet de thèse a été développé autour de deux axes de recherche. (I) Dans un premier temps, les nanoparticules de gemcitabine-squalène ont été fonctionnalisées par un peptide capable de reconnaître et de cibler spécifiquement les cellules cancéreuses pancréatiques. (II) Le deuxième axe de recherche a visé l’encapsulation d’un second principe actif au sein des nanoparticules de gemcitabine-squalène afin de développer le concept de nanoparticule « multi-thérapeutique ». / Pancreatic cancer is a lethal disease with the worst prognosis among all solid tumors. In the last decades, progresses in pancreatic cancer therapy had remained exceedingly slow and disappointing offering minimal benefits in median survival which remains of less than 6 months and the maximum of 5 years in the 6% of patients. One of the major requirements for a successful cancer therapy is its ability to selectively kill cancer cells with minimal damage to healthy tissues. In this context, a great deal of attention focused on advanced nanoscale systems (i.e., nanomedicines) with the aim to overcome the limits associated to the traditional drug delivery modalities. Nanomedicines can indeed enhance drug properties by (i) offering protection from degradation, (ii) enabling controlled release and distribution and increasing bioavailability while reducing undesired side effects.In the current work we aimed to propose novel nanoscale-based strategies to optimize pancreatic cancer treatment taking into account the specific physio-pathology of this tumor. The first approach relied on the design of a targeted nanomedicine able to specifically bind receptors mainly expressed onto pancreatic cancer cells in order to selectively increase drug accumulation in these cells saving healthy ones.In a second approach, by combining two therapeutic agents in the same nanoparticle we constructed a multi-therapeutic drug delivery system capable to increase the therapeutic index of the combined therapy. In particular, taking advantages from the “squalenoylation prodrug approach”, the research activity of this Ph.D. work lead to the to design of (i) a novel peptide-functionalized squalenoyl gemcitabine nanoparticle and (ii) a tyrosine kinase inhibitor-loaded squalenoyl gemcitabine nanoparticle. Obtained nanoparticles were investigated with respect to their physico-chemical properties and in vitro antitumor activity. The efficacy of peptide-functionalized nanoparticles in impairing tumor growth was assessed in vivo on an experimental model of pancreatic cancer.

Syfte: Syftet med detta arbete är att öka förståelsen för hur företag arbetar med strategi och taktik för att skapa förväntan på marknader innan en produkts premiär. Hur det går till när förväntan skapas innan produktpremiär, utan att det slår över och får negativa konsekvenser för produkten eller tjänsten, är ett område som kan anses vara outforskat. Området är en viktig källa till framgång eller fall för marknadsföring och lansering av nya produkter, vilket gör det relevant och intressant, både teoretiskt och praktiskt. Metod: I denna studie har vi använt oss av en kvalitativ forskningsmetod, semistrukturerade intervjuer. Respondenterna är marknadsförare och produktansvariga med en bred bakgrund från olika typer av företag. De var hälften kvinnor, hälften män i olika åldrar. Vid analyseringen användes tematisk analys, då vi kodade intervjuerna för att sedan bilda teman. Resultat & slutsats: Resultatet som har framkommit i denna studie är att företagen lägger stor vikt vid att skapa och förankra planer internt och hos samarbetspartners innan en produktlansering. Samtidigt måste företaget arbeta med förståelse för och val av målgrupp, för att med dessa bitar på plats kunna leverera information till konsumenterna som skapar förväntan på marknader. Om någon del inte är välgjord ökar riskerna till att det blir komplikationer för lanseringen och uppbyggd förväntan riskerar att sänkas. Är grundarbetet välgjort har företagen goda förutsättningar att skapa förväntan. Sedermera är arbetet med att skapa förväntan en process som startar långt innan premiär. Examensarbetets bidrag: Kunskapsbidraget för denna studie är en unik insikt och översikt över hur företag arbetar med strategi och taktik för att skapa förväntan på marknader innan en produkts premiär. Förslag till fortsatt forskning: Ytterligare undersöka konsumenternas perspektiv om detta ämne och deras vilja att använda deras röst. Undersöka hur negativ buzz i slutändan kan bidra till produktframgång. Undersöka skillnaderna mellan tillverkare, grossister och butik när det kommer till kompetens och förståelse för strategier för att skapa förväntan genom sociala medier. / Aim: The purpose of this paper is to increase understanding of how companies work with strategy and tactics to create anticipation in markets. The way in which anticipation is created before the product premiere, without it having a negative impact on the product or service success, is an area that can be considered unexplored. The area is an important source of success or failure for marketing and launching new products, which makes it relevant and interesting, both theoretically and practically. Method: In this study we have used a qualitative research method, semi-structured interviews. The respondents are marketers and product managers with a broad background from different types of companies. They were an equal proportion of women and men of different ages. Thematic analysis was used for coding the interviews and then forming themes. Result & Conclusions: The result that has emerged in this study is that the companies put great emphasis on creating and establish plans internally and with partners before a product launch. At the same time, the organization must work to create an understanding of and choose a target group, in order to be able to supply information to consumers who create anticipations in markets. If any part is not well done, the risk increases that there will be complications for the launch and built-up anticipation risks being lowered. If the groundwork is well done, the companies have good opportunities to create anticipations. Later on, the work of creating anticipation is a process that starts long before the premiere. Contribution of the thesis: The contribution for this study is a unique insight and overview of how companies work with strategy and tactics to create anticipation in markets before a product's premiere. Suggestions for future research: Further explore consumer perspective on this topic and their willingness to use their voice. Examine how negative buzz can ultimately contribute to product success. Examine the differences between manufacturers, wholesalers and retailers when it comes to skills and understanding of strategies to create anticipation.

Os tumores de musculatura liso do útero se desenvolvem a partir do miométrio e podem apresentar carcterísticas clínicas malignas e benignas. Dentre eles, o leiomiossarcoma (LMS) é o tumor maligno mais comum, com altas taxas de metástase e recidiva, mesmo sendo diagnosticado em estágios iniciais. Já os leiomiomas (LM) são os tumores benignos mais frequentes em mulheres em idade reprodutiva. Ambos possuem mesma diferenciação celular, porém com comportamentos clínico e biológico bastante distintos, e até o momento não se dispõe de tratamento específico ou curativo. Nesse contexto, a busca por novos alvos moleculares pode contribuir não só para um melhor entendimento dessas neoplasias, como também para a descoberta de novas terapias. Em estudo prévio foi observada a expressão aumentada de FOXO3a nos sarcomas uterinos, em comparação aos LMs e ao miométrio adjacente (MM). Além disso, sua expressão foi crescente de acordo com o potencial de malignidade do tumor. Assim, o objetivo do presente estudo foi avaliar in vitro o efeito de terapia alvo específica para FOXO3a em células de LM e LMS. Para isto, linhagens celulares de MM (ATCC PCS-460-011), LM (THESCs - CRL-4003) e LMS (SK-UT-1 - HTB-114) foram caracterizadas quanto à expressão basal de FOXO3a (gene e proteína) e submetidas a tratamento com Genisteína e Metformina ou inativação do gene por siRNA. Os efeitos dos tratamentos foram avaliados por PCR em tempo real, Western Blot, imunocitoquímica, ensaios de proliferação, migração e apoptose. Nossos resultados mostraram que todos os tratamentos realizados interferiram na capacidade de proliferação e migração das células, com maior inibição após as 48 horas nos LMS e 72h nos LM. O efeito obtido na transfecção com siRNA apresentou maior eficiência após 48 h da transfecção nos LMS e 72h nos LM. Os efeitos da inibição de FOXO3a foram maiores na proliferação e migração dos LM, porém os resultados não foram estatisticamente significativos. Dentre as substâncias testadas, a Metformina apresentou maior efeito sobre a proliferação, migração e viabilidade das linhagens celulares. A Genisteína também apresentou efeito inibitório nas células, porém o controle com veículo também apresentou o mesmo efeito citotóxico. De modo geral, os efeitos obtidos com os fármacos, foram tempo e concentração dependentes. Em conjunto, nossos resultados sugerem um relevante do FOXO3a nos tumores de musculatura lisa uterinos, além de apresentá-lo como potencial alvo para terapia específica / Smooth muscle tumors of the uterus develop from the myometrium and may present benign and malignant clinical features. Among them, leiomyosarcoma (LMS) is the most frequent malignant tumor, with high rates of metastasis and relapse, even when diagnosed in early stages. On the other hand, leiomyomas (LM) are the most frequent benign tumors in women of reproductive age. Both have the same cellular differentiation, but with very different clinical and biological behaviors, and so far no specific or curative treatment is available. In this context, the search for new molecular targets can contribute not only for a better understanding of these neoplasms, but also for the discovery of new therapies. In a previous study, increased expression of FOXO3a in uterine sarcomas was observed, compared to LMs and adjacent myometrium (MM). In addition, its expression was increasing according to the malignancy potential of the tumor. Thus, the aim of the present study was to evaluate in vitro, the effect of specific targeted therapy for FOXO3a on LM and LMS cells. For this, MM (ATCC PCS-460-011), LM (THESCs-CRL-4003) and LMS (SK-UT-1-HTB-114) cell lines were characterized for basal expression of FOXO3a (gene and protein) and subsequently submitted to treatment with metformin and genistein, or silencing of FOXO3a by siRNA. The effects of the treatments were evaluated by real-time PCR, Western Blot, immunocytochemistry, proliferation, migration and apoptosis assays. Our results showed that all treatments interfered in the proliferation and migration capacity of the cells, with greater inhibition after 48 hours for LMS and 72 hours LM. The effect obtained in the transfection with siRNA showed higher efficiency after 48 hours of transfection in LMS and 72 hours in LM. The effects of inhibition of FOXO3a were greater in the proliferation and migration of the LM, but the results were not statistically significant. Among the substances tested, Metformin had a greater effect on proliferation, migration and viability of the cell lines. Genistein also had an inhibitory effect on the cells, but the control with the vehicle also presented the same cytotoxic effect. In general, the effects obtained with the drugs were time and concentration dependent. Together, our results suggest a relevant role of FOXO3a in uterine smooth muscle tumors, in addition to presenting it as a potential target for specific therapy