Les proteines ErbBjHER appartiennent a la famille des recepteurs a activite tyrosine kinase (RrK). Ces recepteurs participent a un reseau complexe d'jnteractions a la surface de la cellule et controlent la proliferation et la differenciation cellulaire. Cette famille de 4 membres (ErbBljEGFR, ErbB2, ErbB3, ErbB4) forme des homodimeres etjou des heterodimeres apres fixation du ligand. La deregulation de ce reseau d'interactions est associee a de nombreux cancers humains.<br />Le domaine tr~smembranaire joue un role dans la fonction de ces recepteurs et l'implication des motifs de type GxxxG dans les processus d'association suscite un interet particulier. Des etudes experimentales montrent une correlation entre la hierarchie des interactions des recepteurs entiers et celIe des domaines transmembranaires.<br />Une methode theorique de recherche de modeles d'association des heterodimeres a ete develoHpee permettant de quantifier les interactions entre les domaines membranaires et de definir Ie role des motifs dans l'association. Des simulations de dynamique moleculaire effectuees sur l'ensemble des modeles montrent une preference d'association gauche des helices. Les petits residus appartenant aux motifs de dimerisation sont presents a l'interface des deux helices. Nos etudes montrent que les domaines transmembranaires participent a la specificite et a la selectivite des recepteurs dans les processus de dimerisation.

La partie orientale de la Méditerranée, est une des régions sismiques les plus actives en Europe. La présence dans cette région de nombreux lacs et bassins marins isolé fait de cette région un endroit idéal pour étudier l'enregistrement de l'activité sismo- tectonique et les principaux changements climatiques dans des environnements subaquatiques. Dans cette thèse, nous nous intéressons à l'étude de l'enregistrement paléosismique subaquatique du Quaternaire tardive dans deux bassins: la Mer de Marmara et le Golfe de Corinthe. Ces derniers sont des bassins marins isolés traversés par des failles majeures (par exemple, la faille Nord Anatolienne). Nous nous sommes concentrés sur l'identification des traces laissées par des paléoséismes dans l'enregistrement sédimentaire, en particulier, les couches nommées ici: "homogenites + turbidites" (HmTu), lequel représentent des événements sédimentaires quasi instantanés induits par des secousses sismiques. Leur identification a été réalisée à partir des analyses sédimentologiques classiques et des propriétés magnétiques, en particulier, l'anisotropie de la susceptibilité magnétique. Les analyses ont été effectuées sur les sédiments carottés à l'aide du système géant à piston CALYPSO. Ces sédiments représentent les dernières 17 cal ka BP de sédimentation, et ont enregistré le dernier passage de conditions non marine à des conditions marines. Dans la mer de Marmara, ce passage a été identifié autour de 12,8 cal ka BP, tandis que, dans le golfe de Corinthe était d'environ 11,7 cal ka BP. Dans les deux bassins, l'enregistrement des événements instantanés considérés comme induits par des séismes a permis l'estimation d'un intervalle moyen de récurrence. Dans le bassin de Çınarcık de la mer de Marmara, l'intervalle de récurrence moyenne minimale varie entre ~155 et ~365 ans, tandis que pour la partie orientale du golfe de Corinthe, l'intervalle de récurrence moyenne minimale varie entre ~400 et ~500 ans. Enfin, dans le bassin central de la mer de Marmara, l'étude des événements HmTu corrélées sur les côtés opposés d'un segment de la faille nord-anatolienne permet d'estimer le décalage vertical co-sismique pour une période continue de 2 ka. Des valeurs significatives observées étaient des décalées verticalement (jusqu'à 144 cm), ce qui implique un déplacement verticalement dominant (normal) pour ce segment. / The Eastern Mediterranean is one of most seismically active and rapidly deforming regions in Europe. The presence in this region of numerous lakes and isolate marine basins makes this region an ideal area to study the record of the seismo-tectonic activity and the main climatic changes in sub-aquatic environments. In this thesis we are interested in studying the Late Quaternary sub-aquatic paleoseismic record of two basins: The Sea of Marmara and The Gulf of Corinth. Both are isolated deep marine basins, crossed by major faults (e.g. the North Anatolian Fault). We focused on identifying the traces left by paleoearthquakes in the sedimentary record, particularly, the layers named here: “homogenites+turbidites” (HmTu), which represent individual sedimentary seism-induced events. Their identification was performed through classical sedimentological analyzes and magnetic properties, in particular the Anisotropy of Magnetic Susceptibility. Analyzes were conducted on sediment recovered by long Calypso piston cores. These sediments represent the last 17 cal kyr BP of sedimentation, and have recorded the last non-marine to marine change. In the Sea of Marmara, this passage was identified around 12.8 cal kyr BP, whereas in the Gulf of Corinth was around 11.7 cal kyr BP. In both basins, the record of the earthquake induced instantaneous events allowed the estimation of the average earthquake recurrence interval. In the Çinarcik Basin of the Sea of Marmara, the minimum average recurrence interval varies between ∼155 and 365 yr, while for the eastern part of the Gulf of Corinth the minimum average recurrence interval varies between ∼400 and ∼500 yr. Finally, in the Central Basin of the Sea of Marmara, the study of correlated HmTu events at opposite sides of a segment of the North Anatolian Fault allows the estimation of the co-seismic vertical offset for a continuous period of 2 kyr. Significant values of vertical offset were observed (up to 144 cm), implying a dominant vertical (normal) throw for this fault segment.

Today, GNSS (Global Navigation Satellite Systems) is a widely spread technology and are used in many different ways. GNSS is the collective name for American GPS, Russian Glonass and European Galileo and some other smaller navigation systems. When surveying with GNSS technology, there are many things that can affect the accuracy of the measurements. In this thesis the focus is on how these measurements are being effected by disturbances in the ionosphere. The conditions in the ionosphere are largely affected by the amount of solar activities. A guideline for the amount of solar activity is to monitoring the sunspot numbers and the solar cycle. With more radiation occurring at radio frequencies as well as in the UV-region with more sunspots. The resulting free electrons influence the GNSS signals as they propagate through the ionosphere and the ionized gases cause the phase of the GNSS signal to shift and slows down the speed of the signal.   By processing data from three SWEPOS stations forming one shorter and one longer baseline, the estimated errors for the measurements are being calculated for different conditions in the ionosphere. The focus in this thesis is at the accuracy and precision of those measurements. To capture different ionospheric conditions, a few days from different parts of the solar cycle are chosen as representation for these different conditions. For each day different times of day and different length of measuring sessions are being calculated. All the calculations of the baselines are carried out in Trimble Business Center, and the results are presented in charts for each day and some key measurements are presented for the whole studied period.   The result gives no clear correlation between the number of sunspots and the accuracy and precision of the GNSS measurements, and it is not possible from the result to see that a particular time of the day is better suited for executing GNSS surveying.   From the studied material and the result of this thesis it is hard to see any dependency between the solar activity and the accuracy on GNSS measurements. It is not possible from the result see any connection between the length of the measuring time and the outcome of the measurement, as it is often no clear pattern stating that a longer measuring time would give better measurement.   The results differ in many ways from other reports on this subject and the result are so totally different that something seems to not be right in this report. If a correlation analysis had been performed instead of just an ocular examination, it had probably given a more satisfying result of the studied material.   The result of this thesis is in many ways on collision course with other similar report on this subject and therefore the results in this report can be neglected and rejected in favor of the result of this other more scientific reports.

Cette thèse porte sur les formes d'engagement et des revendications de légitimité affirmés au tour des organisations de desocupados. Elle compare des militants avec les non militants observés dans leur même milieu social. Elle analyse deux mouvements de travailleurs desocupados (MTD) situés dans les villes de Berisso et Berazategui entre 2003 et 2007. Cette interrogation s'appuis sur une analyse ethnographique des liens établis entre membres et non membres, notamment parmi leurs proches et vecinos. Les parcours sont perméables sur le long terme entre engagement et retrait, division du travail entre familiers pour répartir les responsabilités privées, économiques et publiques. L'analyse porte sur le mouvement piquetero en tant qu'acteur politique, mais aussi, sur le mode de vie, les conceptions de l'activité et de la justice mises en œuvre par l'engagement dans ces organisations. Une première partie montre la manière dont les actions politiques des mouvements de desocupados sont imbriquées dans le vécu quotidien des acteurs. Se dégagent au fils des observations, des moments militants dans les milieux sociaux concernés. La seconde partie analyse la notion de travail et de l'activité étendue, également, à l'activité politique. Le travail demeure au centre des représentations de l'activité légitime, mais recouvre de nouvelles pratiques et de nouvelles formes d'activité radicalement opposées à la seule loi du marché. " Vivre en travaillant " peut devenir " vivre des productivos ". Enfin, le rapport à la parole des membres et non membres au sein du groupe signale une forme de publicité faite d'une confiance mutuelle, provisoire, qui étend la délibération aux lieux de travail.

L'objectif de cette recherche était d’étudier l’activité collective dans les classes de collèges ÉCLAIR en EPS lors d'un dispositif de travail par ateliers en gymnastique avec des enseignants "réussissants". Dans ces classes, une des difficultés majeures pour les enseignants est la mise au travail durable des élèves. Au cours de la leçon, l’activité collective est particulièrement instable en raison d'une activité des élèves marquée par de nombreux décrochages, un refus de travailler, une agitation continue pouvant aller jusqu’à la violence. Toutefois, certains enseignants "réussissants" parviennent à faire travailler le collectif d’élèves de façon relativement durable au cours de la leçon. Cette recherche a donc analysé le travail collectif dans les classes avec ces enseignants pour identifier sa forme typique et comprendre les processus à partir desquels il se construisait. Elle a été conduite dans le cadre théorique et méthodologique du Cours d’action, permettant d’appréhender l'activité collective à partir d'une entrée par l’activité et le sens pour les acteurs. Les études de cas se sont déroulées dans des collèges ÉCLAIR de la banlieue lilloise, en EPS, lors de 27 leçons de gymnastique par ateliers. Sept enseignants "réussissants" et 37 élèves ont été filmés puis amenés à exprimer leur expérience vécue en classe lors d'entretiens d’autoconfrontation. L’activité collective a été renseignée à deux niveaux d’analyse autonomes et interdépendants : d’une part, celui de l’activité individuelle-sociale de l’enseignant et des élèves par croisement des matériaux relatifs à la description des comportements observés en classe et ceux relatifs aux verbatim d’entretien ; et d’autre part au niveau de la configuration de l’activité de la classe en repérant les points d'articulation entre l'activité de l'enseignant et celle des élèves et en identifiant la forme de l’activité collective qui en émerge. Les résultats montrent que malgré une constante agitation dans les ateliers de gymnastique, une activité collective de travail domine dans la classe sans qu’une perturbation majeure vienne rompre les leçons étudiées. Premièrement, l'analyse de l’activité individuelle-sociale a fait ressortir deux traits typiques : une forme bipolaire organisée par la dyade « travail – jeu » pour les élèves et par la dyade « instruction – contrôle » pour l’enseignant. Deuxièmement, la configuration d’activité collective en classe a une forme studieuse typique : elle repose sur des processus d'ostension et de masquage permettant l'articulation des activités de l'enseignant et des élèves dans la classe. Les conclusions de cette étude permettent de répondre à des visées épistémiques concernant la compréhension de l'activité collective et de sa construction, ainsi que des visées transformatives apportant des éclairages sur les pratiques d’apprentissage et d’enseignement dans les classes issues de milieux « difficiles ». / The objective of this research was to study collective activity in gymnastics workshops taught by “successful” physical education teachers in middle schools benefiting from compensatory education policies (ÉCLAIR). One of the major difficulties for teachers in these classes is to obtain sustained effort from the students. Group work is particularly unstable during lessons because the students’ individual activity is characterized by frequent disengagement, refusal to work, and continuous agitation that may take a violent turn. Certain teachers nevertheless succeed in getting their students to work in a relatively durable manner over the course of the lesson. This work thus analyzed the collective activity in classes with these teachers to identify its typical form and to understand the processes by which it was constructed. The theoretical and methodological framework for this research was the course of action, which is well suited to the study of collective activity as it specifically targets activity and the meaning it has for the actors. The case studies were conducted in middle schools in the Lille region, both part of the ÉCLAIR program, during 27 physical education classes organized as gymnastics workshops. Seven “successful” teachers and 37 students were filmed and then asked to describe their lived experiences in the classroom during self-confrontation interviews. The group activity was analyzed from two autonomous and interdependent perspectives: first, we analyzed the individual-social activity of the teachers and students by crossing the descriptive data on the behaviors observed in class with the verbatim data from the self-confrontation interviews and, second, we analyzed the configuration of the classroom activity by identifying the points of articulation between the teacher’s and students’ activity and then identifying the form of collective activity that emerged. The results showed that, despite constant agitation, the collective activity in these workshops was dominated by group work with no major disruptions in the lessons. The analysis of individual-social activity revealed two typical forms: a bipolar form organized by the "work-play" dyad for the students and by the "instruction-control" dyad for the teacher. The configuration of classroom activity also showed a form typical to classroom situations: it was based on processes of revealing and masking that made the articulation of teachers’ and students’ activities possible. The findings of this research enhance our understanding of collective activity and how it is constructed and provide greater insight into the learning practices of students from “difficult” backgrounds and the teaching practices of teachers who operate successfully in these classrooms.

Nesta tese serão discutidos alguns aspectos químicos, fotoquímicos e fotofísicos importantes no desenvolvimento de fotossensibilizadores para aplicação em terapia fotodinâmica (TFD). Os estudos abrangeram a investigação de 29 espécies, sendo que 20 delas continham um anel porfirínico convencional e 9 eram porfirinas duplamente N-confusas. As primeiras foram sintetizadas para apresentarem diferentes números e tipos de substituintes catiônicos coordenados à posição meso do anel porfirínico, mais especificamente as meso(N-4-piridil)fenilporfirinas e as meso(N-3-piridil)fenilporfirinas contendo um, dois (em trans), dois (em cis), três e quatro grupos [Ru(bipy)2Cl]+ ou CH3+ (figura 1). Em contraste com as porfirinas convencionais. as outras 9 estruturas consistiram das formas protonadas, neutras e desprotonadas de porfirinas que exibem dois anéis pirrólicos adjacentes voltados para fora do anel porfirínico, mais especificamente a base livre (H2N2CP) e os respectivos complexos de prata (AgHN2CP) e de cobre (CuHN2CP). Esta peculiaridade estrutural permite coordenar metais de transição por meio de dois átomos de carbono e dois de nitrogênio, estabilizando fortemente íons metálicos em estados de oxidação anormalmente elevados como Ag3+ e Cu3+. Os dois nitrogênios pirrólicos externos ao anel são susceptíveis tanto a protonação quanto a desprotonação, possibilitando a modulação das propriedades eletrônicas desses compostos por simples alteração no pH do meio. De um modo geral, a acidificação das amostras perturbou mais significativamente suas propriedades fotoquímicas que a desprotonação. Em especial, as espécies neutras metaladas apresentaram bandas bastante intensas na região fototerapêutica de 600 a 750 nm, além de um elevado rendimento quântico de formação de oxigênio singlete (&#934;&#916; &#62; 0,90 para o complexo Ag(III)). Todavia, um processo de fotodecomposição atribuído ao ataque do 1O2 formado, também foi observado. Ambos os processos, &#934;&#916; e fotodecomposição, parecem ser modulados pelo metal coordenado no centro do anel. Tanto a coordenação de Ag(lII) como Cu(lII) aumentou o &#934;&#916;, provavelmente devido ao efeito do metal pesado sobre o cruzamento interssistema. Todavia, verificou-se que o derivado AgHN2CP é muito menos reativo que o CuHN2CP e a base livre. Os motivos deste comportamento ainda não são completamente compreendidos, mas é evidente que o complexo de Ag(lII) apresenta melhores propriedades fotodinâmicas que as demais. Em comparação com as porfirinas duplamente N-confusas, as demais 20 porfirinas catiônicas são muito mais solúveis em água. Neste caso, o enfoque dos experimentos foi direcionado às aplicações biológicas propriamente ditas. Assim, além das propriedades fotofísicas associadas à formação de oxigênio singlete (1O2), também foram exploradas a influência do número dos substituintes periféricos sobre a ligação e danos fotooxidativos provocados em eritrócitos, lipossomos, mitocôndrias e células cancerígenas. Os resultados de &#934;&#916; mostraram ser inversamente proporcionais ao número de cargas positivas, para as porfirínas contendo grupos N-4metilpiridíniom. Estas diferenças foram consistentes com os efeitos de agregação, causados pela formação de dímeros ou oligômeros, no caso das espécies mais hidrofóbicas. Os coeficientes de partição em n-octanol/água (logPOA) corroboraram estes resultados, apresentando uma dependência inversamente proporcional ao número de cargas positivas ou proporcional ao número de resíduos fenila. As eficiências de ligação à bicamada lipídica mitocondrial, lipossomal e celular e os processos de peroxidação lipídica fotoinduzidos mostraram uma dependência proporcional aos valores de logPOA, exceto para as porfirinas contendo complexos de rutênio. O comportamento não linear neste último caso pode estar associada a dissociação da ligação rutênio-porfirina, devido a elevada força iônica das soluções fisiológicas utilizadas. Neste caso, a descoordenação dos complexos periféricos deve promover uma drástica diminuição da solubilidade do composto. Por este motivo os experimentos foram conduzidos somente com as espécies metiladas. No caso, de mitocôndrias isoladas verificou-se que a interação também apresenta uma razoável participação do potencial de membrana. De fato, quando a mitocôodria estava energizada foi observado um acréscimo de 15% no acúmulo da 3P2cMe em comparação com à mitocôndria desacoplada. Para as demais porfirinas houve um menor acúmulo com o aumento do número de resíduos N-3-metilpirídínio. Em todos os experimentos, um comportamento diferenciado foi observado no caso dos isômeros dicatiônicos. A espécie com cargas dispostas nas posições cis (3P2cMe) apresentou praticamente o dobro de eficiência que o correspondente isômero trans (3P2tMe). Este comportamento parece consistente com uma maior penetração da primeira espécie na bicamada lipídica, proporcionado pelo caráter antipático de sua estrutura química. Os resultados apresentados neste estudo mostraram que estas séries de meso-porfirinas catiônicas constituem sistemas bastante interessantes para a compreensão dos efeitos estruturais envolvidos na ação fotodinâmica dos compostos sobre membranas lipídicas, devido à possibilidade de modulação da relação hidrofilicidade/lipofilicidade, sem alterar significativamente a eficiência na formação de 1O2. Além disso, a disposição espacial das cargas positivas na estrutura do fotossensibilizador demonstrou ser extremamente crítica no processo de ancoragem do mesmo na membrana celular ou mitocondrial e consequentemente em causar efeitos fotooxidativos mais eficientemente. As porfirinas contendo grupos 3-metilpiridínio apresentam vantagens sobre as 4-metilpiridínio, normalmente estudas, por serem mais hidrofílicas, terem menor tendência de agregação e possuírem similar atividade fotodinâmica. Em especial, a estrutura anfipática da 3P2cMe associada a sua elevada formação de 1O2 e estabilidade lhe conferem uma interessante potencialidade como agente fototerapêutico. / In this thesis the chemical, photochemical and photophysical relevant aspects for the development of new photosensitizers for photodynamic therapy applications are discussed. 20 conventional porphyrins species and 9 doubly Nconfused porphyrins species were investigated. The first series contains variable number (1 to 4) of cationic groups, [Ru(bipy)2CI]+ or CH3+ (figure 1), bound to the meta or para-pyridil N-atoms of meso-phenylpyridylporphyrins. On the other hand, the protonated, neutral and deprotonated doubly N-confused porphyrins possess two rotated adjacent pyrrol rings, such that they have two coordinating camon atoms. Consequently, transition metal ions in unusually high oxidation states such as Ag(lII) and Cu(lII) can be stabilized by this structure, while the outer pyrrol N-atoms are susceptible to protonation and deprotonation reactions, allowing the modulation of their electronic properties simply by controlling the pH of the solution. In general, the protonation perturbed more significantly the photochemical properties than the deprotonation. The neutral species exhibit intense absorption bands in the phototherapeutical range (600 to 750 nm), associated with a high singlet oxygen sensitization quantum yield (&#934;&#916;&#62; 0,90 for the Ag(lII) complex). Furthermore, a photodecomposition process due to the reaction with 1O2 was also identified. Both &#934;&#916; and photodecomposition are influenced by the metal ion coordinated to the doubly N-confused porphyrin ring. The coordination of Ag(lIl) and Cu(lII) increased &#934;&#916;, probably due to the enhancement of intersystem crossing quantum yield associated with the heavy atom effects. However, AgHN2CP is much more stable than CuHN2CP or the free-base, but the reasons are not clear and should be further investigated. The above results clearly evidence the superior properties of the Ag(lIl) complex as photosensitizer for PDT applications. The 20 cationic pyridylporphyrin derivatives are much soluble in water and experiments directed to biological applications could be carried out. Accordingly, in addition to the quantum yield for singlet oxygen generation(1O2), the effect of the stereochemistry and number and position of the electrically charged substituents on the binding constants and photooxidative damage on erythrocytes, lipossomes, mitochondria and HeLa cells were evaluated. The &#934;&#916; values were constant in the case of the N-3-methylpyridinium derivatives but inversely proportional to the number of positive charges for the N-4-methylpyridinium derivatives. This was assigned to an increase of aggregation of the sensitizers as the number of meso-phenyl rings and the lipophylicity increase. The partition coefficients in n-octanol/water (logPOA) corroborate that assertion showing a linear correlation with increasing of the number of phenyl groups. The binding efficiency towards the mitochondrial, lipossomal and cellular membrane and the photo-induced lipidic peroxidation processes were directly proportional to the logPOA, except for the ruthenated porphyrins. This last behavior may be associated with the dissociation reactions of the rutheniumpyridylporphyrin bond in the physiologic solution. Accordingly, those experiments were carried out only with the methylpyridynium derivatives. For the first time, we showed inequivocally that the interaction of the methyllated porphyrins with mitochondria are significantly influenced by the membrane potential. In fact, when the mitochondria was energized a 15% increase was observed in the binding constants of 3P2cMe in comparison with teh results with decoupled mitochondria. Also, the amount of bond porphyrin sensitizer decreased as the number of methylpyridynium groups was increased, following the same tendency as above. The behavior of the cis (3P2cMe) and trans (3P2tMe) isomers didn\'t follow the general tendency for the other derivatives in a series, such that the binding constants of the cis isomers were always about twice higher than for the trans, which were much higher than predicted by the general behavior. This means that they can associate more strongly and penetrate deeper in the membrane than the more charged porphyrin derivatives. In particular, the cis species is amphiphilic, i.e. possess an adequate structure for that interaction. The results presented in this thesis showed conclusively that the two series of meso-phenylpyridylporphyrins are adequate for the investigation of the effect of the stereochemistry on the bonding ability and photodynamic properties of those photosensitizers. This is particularly true due to the possibility of modulating the ratio between hydrophobicitylhydrophylicity and the stereochemistry without influencing significantly the quantum yield for 1O2 generation. The meta- series showed an advantage over the conventionally used para- series of methylpyridynium porphyrins because it is more soluble and shows lesser tendency to aggregate. In condusion, the amphiphilic 3P2cMe is the species with the highest potentiallity as por sensitizer because of its high binding constants, low tendency to associate and high &#934;&#916;.

Nesta tese serão discutidos alguns aspectos químicos, fotoquímicos e fotofísicos importantes no desenvolvimento de fotossensibilizadores para aplicação em terapia fotodinâmica (TFD). Os estudos abrangeram a investigação de 29 espécies, sendo que 20 delas continham um anel porfirínico convencional e 9 eram porfirinas duplamente N-confusas. As primeiras foram sintetizadas para apresentarem diferentes números e tipos de substituintes catiônicos coordenados à posição meso do anel porfirínico, mais especificamente as meso(N-4-piridil)fenilporfirinas e as meso(N-3-piridil)fenilporfirinas contendo um, dois (em trans), dois (em cis), três e quatro grupos [Ru(bipy)2Cl]+ ou CH3+ (figura 1). Em contraste com as porfirinas convencionais. as outras 9 estruturas consistiram das formas protonadas, neutras e desprotonadas de porfirinas que exibem dois anéis pirrólicos adjacentes voltados para fora do anel porfirínico, mais especificamente a base livre (H2N2CP) e os respectivos complexos de prata (AgHN2CP) e de cobre (CuHN2CP). Esta peculiaridade estrutural permite coordenar metais de transição por meio de dois átomos de carbono e dois de nitrogênio, estabilizando fortemente íons metálicos em estados de oxidação anormalmente elevados como Ag3+ e Cu3+. Os dois nitrogênios pirrólicos externos ao anel são susceptíveis tanto a protonação quanto a desprotonação, possibilitando a modulação das propriedades eletrônicas desses compostos por simples alteração no pH do meio. De um modo geral, a acidificação das amostras perturbou mais significativamente suas propriedades fotoquímicas que a desprotonação. Em especial, as espécies neutras metaladas apresentaram bandas bastante intensas na região fototerapêutica de 600 a 750 nm, além de um elevado rendimento quântico de formação de oxigênio singlete (&#934;&#916; &#62; 0,90 para o complexo Ag(III)). Todavia, um processo de fotodecomposição atribuído ao ataque do 1O2 formado, também foi observado. Ambos os processos, &#934;&#916; e fotodecomposição, parecem ser modulados pelo metal coordenado no centro do anel. Tanto a coordenação de Ag(lII) como Cu(lII) aumentou o &#934;&#916;, provavelmente devido ao efeito do metal pesado sobre o cruzamento interssistema. Todavia, verificou-se que o derivado AgHN2CP é muito menos reativo que o CuHN2CP e a base livre. Os motivos deste comportamento ainda não são completamente compreendidos, mas é evidente que o complexo de Ag(lII) apresenta melhores propriedades fotodinâmicas que as demais. Em comparação com as porfirinas duplamente N-confusas, as demais 20 porfirinas catiônicas são muito mais solúveis em água. Neste caso, o enfoque dos experimentos foi direcionado às aplicações biológicas propriamente ditas. Assim, além das propriedades fotofísicas associadas à formação de oxigênio singlete (1O2), também foram exploradas a influência do número dos substituintes periféricos sobre a ligação e danos fotooxidativos provocados em eritrócitos, lipossomos, mitocôndrias e células cancerígenas. Os resultados de &#934;&#916; mostraram ser inversamente proporcionais ao número de cargas positivas, para as porfirínas contendo grupos N-4metilpiridíniom. Estas diferenças foram consistentes com os efeitos de agregação, causados pela formação de dímeros ou oligômeros, no caso das espécies mais hidrofóbicas. Os coeficientes de partição em n-octanol/água (logPOA) corroboraram estes resultados, apresentando uma dependência inversamente proporcional ao número de cargas positivas ou proporcional ao número de resíduos fenila. As eficiências de ligação à bicamada lipídica mitocondrial, lipossomal e celular e os processos de peroxidação lipídica fotoinduzidos mostraram uma dependência proporcional aos valores de logPOA, exceto para as porfirinas contendo complexos de rutênio. O comportamento não linear neste último caso pode estar associada a dissociação da ligação rutênio-porfirina, devido a elevada força iônica das soluções fisiológicas utilizadas. Neste caso, a descoordenação dos complexos periféricos deve promover uma drástica diminuição da solubilidade do composto. Por este motivo os experimentos foram conduzidos somente com as espécies metiladas. No caso, de mitocôndrias isoladas verificou-se que a interação também apresenta uma razoável participação do potencial de membrana. De fato, quando a mitocôodria estava energizada foi observado um acréscimo de 15% no acúmulo da 3P2cMe em comparação com à mitocôndria desacoplada. Para as demais porfirinas houve um menor acúmulo com o aumento do número de resíduos N-3-metilpirídínio. Em todos os experimentos, um comportamento diferenciado foi observado no caso dos isômeros dicatiônicos. A espécie com cargas dispostas nas posições cis (3P2cMe) apresentou praticamente o dobro de eficiência que o correspondente isômero trans (3P2tMe). Este comportamento parece consistente com uma maior penetração da primeira espécie na bicamada lipídica, proporcionado pelo caráter antipático de sua estrutura química. Os resultados apresentados neste estudo mostraram que estas séries de meso-porfirinas catiônicas constituem sistemas bastante interessantes para a compreensão dos efeitos estruturais envolvidos na ação fotodinâmica dos compostos sobre membranas lipídicas, devido à possibilidade de modulação da relação hidrofilicidade/lipofilicidade, sem alterar significativamente a eficiência na formação de 1O2. Além disso, a disposição espacial das cargas positivas na estrutura do fotossensibilizador demonstrou ser extremamente crítica no processo de ancoragem do mesmo na membrana celular ou mitocondrial e consequentemente em causar efeitos fotooxidativos mais eficientemente. As porfirinas contendo grupos 3-metilpiridínio apresentam vantagens sobre as 4-metilpiridínio, normalmente estudas, por serem mais hidrofílicas, terem menor tendência de agregação e possuírem similar atividade fotodinâmica. Em especial, a estrutura anfipática da 3P2cMe associada a sua elevada formação de 1O2 e estabilidade lhe conferem uma interessante potencialidade como agente fototerapêutico. / In this thesis the chemical, photochemical and photophysical relevant aspects for the development of new photosensitizers for photodynamic therapy applications are discussed. 20 conventional porphyrins species and 9 doubly Nconfused porphyrins species were investigated. The first series contains variable number (1 to 4) of cationic groups, [Ru(bipy)2CI]+ or CH3+ (figure 1), bound to the meta or para-pyridil N-atoms of meso-phenylpyridylporphyrins. On the other hand, the protonated, neutral and deprotonated doubly N-confused porphyrins possess two rotated adjacent pyrrol rings, such that they have two coordinating camon atoms. Consequently, transition metal ions in unusually high oxidation states such as Ag(lII) and Cu(lII) can be stabilized by this structure, while the outer pyrrol N-atoms are susceptible to protonation and deprotonation reactions, allowing the modulation of their electronic properties simply by controlling the pH of the solution. In general, the protonation perturbed more significantly the photochemical properties than the deprotonation. The neutral species exhibit intense absorption bands in the phototherapeutical range (600 to 750 nm), associated with a high singlet oxygen sensitization quantum yield (&#934;&#916;&#62; 0,90 for the Ag(lII) complex). Furthermore, a photodecomposition process due to the reaction with 1O2 was also identified. Both &#934;&#916; and photodecomposition are influenced by the metal ion coordinated to the doubly N-confused porphyrin ring. The coordination of Ag(lIl) and Cu(lII) increased &#934;&#916;, probably due to the enhancement of intersystem crossing quantum yield associated with the heavy atom effects. However, AgHN2CP is much more stable than CuHN2CP or the free-base, but the reasons are not clear and should be further investigated. The above results clearly evidence the superior properties of the Ag(lIl) complex as photosensitizer for PDT applications. The 20 cationic pyridylporphyrin derivatives are much soluble in water and experiments directed to biological applications could be carried out. Accordingly, in addition to the quantum yield for singlet oxygen generation(1O2), the effect of the stereochemistry and number and position of the electrically charged substituents on the binding constants and photooxidative damage on erythrocytes, lipossomes, mitochondria and HeLa cells were evaluated. The &#934;&#916; values were constant in the case of the N-3-methylpyridinium derivatives but inversely proportional to the number of positive charges for the N-4-methylpyridinium derivatives. This was assigned to an increase of aggregation of the sensitizers as the number of meso-phenyl rings and the lipophylicity increase. The partition coefficients in n-octanol/water (logPOA) corroborate that assertion showing a linear correlation with increasing of the number of phenyl groups. The binding efficiency towards the mitochondrial, lipossomal and cellular membrane and the photo-induced lipidic peroxidation processes were directly proportional to the logPOA, except for the ruthenated porphyrins. This last behavior may be associated with the dissociation reactions of the rutheniumpyridylporphyrin bond in the physiologic solution. Accordingly, those experiments were carried out only with the methylpyridynium derivatives. For the first time, we showed inequivocally that the interaction of the methyllated porphyrins with mitochondria are significantly influenced by the membrane potential. In fact, when the mitochondria was energized a 15% increase was observed in the binding constants of 3P2cMe in comparison with teh results with decoupled mitochondria. Also, the amount of bond porphyrin sensitizer decreased as the number of methylpyridynium groups was increased, following the same tendency as above. The behavior of the cis (3P2cMe) and trans (3P2tMe) isomers didn\'t follow the general tendency for the other derivatives in a series, such that the binding constants of the cis isomers were always about twice higher than for the trans, which were much higher than predicted by the general behavior. This means that they can associate more strongly and penetrate deeper in the membrane than the more charged porphyrin derivatives. In particular, the cis species is amphiphilic, i.e. possess an adequate structure for that interaction. The results presented in this thesis showed conclusively that the two series of meso-phenylpyridylporphyrins are adequate for the investigation of the effect of the stereochemistry on the bonding ability and photodynamic properties of those photosensitizers. This is particularly true due to the possibility of modulating the ratio between hydrophobicitylhydrophylicity and the stereochemistry without influencing significantly the quantum yield for 1O2 generation. The meta- series showed an advantage over the conventionally used para- series of methylpyridynium porphyrins because it is more soluble and shows lesser tendency to aggregate. In condusion, the amphiphilic 3P2cMe is the species with the highest potentiallity as por sensitizer because of its high binding constants, low tendency to associate and high &#934;&#916;.

La collaboration MUNU se propose d'etudier la diffusion neutrino-electron avec pour objectif la mesure du moment magntique du neutrino, qui pourrait etre responsable de l'enigme des neutrinos solaires. Le principe de fonctionnement de cette experience, situee aupres d'un reacteur nucleaire du Centre de Production Electrique de Bugey (France), est base sur une Chambre a Projection Temporelle immergee dans huit tonnes de scintillateur liquide agissant comme blindage actif anti-Compton.

La lipase de Yarrowia lipolytica (EC 3.1.1.3) est une enzyme appartenant à la classe des hydrolases. La non pathogénicité et le caractère hyperproducteur en lipase de cette levure lui confèrent une place de choix au sein de lunité de Bio-industries du Centre Wallon de Biologie Industrielle. Ce présent travail sinscrit dans le cadre général du développement industriel de la lipase de Yarrowia lipolytica et concerne plus particulièrement le traitement post-culture de lenzyme afin de réaliser des formes liquides, poudres atomisées, immobilisées et enrobées à laide des polymères acryliques. Latomisation de la lipase en présence ou en absence de poudre de lait a permis lacquisition de poudres fluentes, stables à 4 et 20°C et présentant des températures de transition vitreuse comprises entre 51 et 79°C. Lactivité deau de conservation des poudres était ≤ 0.4. La stabilisation de lenzyme sous forme de liquide concentré réalisée avec le monopropylène glycol (MPG), les inhibiteurs de protéases et lirradiation aux rayons gamma ont révélé que le MPG à 50% et la technique dirradiation au rayon gamma permettaient la stérilisation et la préservation de lactivité enzymatique. Par ailleurs, limmobilisation de cette enzyme par trois techniques (adsorption, inclusion et liaison covalente) a révélé une amélioration de ses propriétés caractéristiques telles que la thermostabilté et la résistance aux solvants. La technique dimmobilisation par adsorption et par liaison covalente a permis une utilisation multiple de lenzyme. Létude préliminaire de faisabilité des formes galéniques à base de la lipase de Y. lipolytica a montré la capacité de cette enzyme à être mise sous forme de comprimés et de poudres encapsulées. La comparaison réalisée in vitro entre le Créon 150mg (produit pharmaceutique) et les formes galéniques à base de la lipase a montré des temps de gastro-résistance et de délitage similaires. Ces différentes formules de la lipase posent des jalons nécessaires pour leurs applications dans des secteurs agroalimentaires, environnementaux et pharmaceutiques. Yarrowia lipolytica lipase (EC.3.1.1.3) is an enzyme which belongs to the class of hydrolases. Nonpathogenicity and the high-lipase producing character of this yeast have emphasised its use within the laboratory of Bio-industry of the Walloon Center of Industrial Biology. The present work lies within the general scope of the industrial development of the lipase from Yarrowia lipolytica. More particularly it relates to the post-culture treatment of the enzyme in order to obtain liquid forms, atomized powders, immobilized and coated enzymes using acrylic polymers. The atomization of lipase in presence or absence of milk powder allowed the achievement of flowing, stable powders at 4 and 20°C, with glass transition temperatures ranging between 51 and 79°C. The water activity of preservation of the powders was ≤ 0.4. Stabilization of the enzyme under the form of concentrated liquid carried out with monopropylen glycol (MPG), proteases inhibitors and gamma irradiation revealed that MPG (50%) and gamma irradiation allowed sterilization and conservation of the enzymatic activity. In addition, the immobilization of the enzyme through three techniques (adsorption, inclusion and covalent bond) revealed an improvement of some properties such as thermostability and resistance to solvents. Immobilization by adsorption and covalent bond allowed multiple uses of the enzyme. The preliminary study of feasibility of galenic forms containing the lipase from Y. lipolytica showed the capacity of this enzyme to be put under the form of tablets and encapsulated powders. The in vitro comparison of Creon 150mg (pharmaceutical product) and galenic forms containing the lipase, showed similar times of acid-resistance and of disintegration. These various formulas of the lipase constitute milestones necessary for their applications in food, environmental and pharmaceutical industries.

Dans cette thèse, j'ai essayé d'identifier le mode d'action de composés biscationiques récemment synthétisés et leurs interactions avec les parasites résistants à la chloroquine. Les activités de divers représentants des composés ammonium bisquaternaire, des amidines alkyl (provenant du groupe de professeur Vial, Montpellier, France), des amidines bisbenzyl (provenant du groupe Chimie de l'Université de Liverpool, Royaume-Uni) ont été comparés à la chloroquine et la pentamidine. Leurs potentiels de résistance croisée avec la chloroquine ont également été étudiés. Dans ce but, deux lignées cellulaires modifiées génétiquement par échange allélique, C3Dd2 et C2GCO3 furent utilisées.Parmi les amidines bisbenzyl, une série de composés appartenant aux guanidines, thiazoles et triazoles ont été criblés pour leur activité contre des souches résistantes et sensibles à la chloroquine chez Plasmodium falciparum. Une hypersensibilité significative est observée pour les amidines bisbenzyl parmi les isolats affichant un PfCRT mutant. Aucune différence n'est observée pour les composés provenant du groupe Vial. Pour comprendre le mode d'action et le rôle de PfCRT, j'ai réalisé des expériences de fixation compétitives (competitive binding') et de cristallisation d'hème. Tous les composés ont montré à différents degrés des interactions avec l'hème, cependant il fut observé que leur activité ne corrélait pas avec l'inhibition de la cristallisation d'hème. Une des raisons possibles à cela est que les différences structurales peuvent jouer un rôle important dans le transport du composé. De plus, j'ai étudié l'effet du pH sur l'activité des composés en utilisant les lignées cellulaires modifiées génétiquement par échange allélique afin d'observer l'effet du gradient de proton sur le transport de la chloroquine et de la pentamidine. Des différences significatives de l'activité de la chloroquine furent observées chez les deux souches. Malgré les valeurs de pKa élevées pour la pentamidine, il y avait une différence significative dans la sensibilité pour ce composé chez les souches quand le pH a changé.Car les diamidines requièrent des transporteurs pour traverser les barrières membranaires et qu'un possible transporteur de choline a été caractérisé chez Plasmodium falciparum, j'ai également réalisé des études initiales sur la caractérisation moléculaire de ce transporteur. Un gène qui encode une protéine chez P. falciparum avec une similarité significative aux eucaryotes supérieurs fut identifié en utilisant des analyses bioinformatiques et fut employé dans une transformation et des études analyses fonctionnelles.En conclusion, ce travail suggère qu'il est possible d'utiliser de nouveaux amidines bisbenzyl pour cibler spécifiquement les souches résistantes à la quinoléine chez Plasmodium falciparum, arborant des allèles de PfCRT mutantes. En adhérant à cette hypothèse et sachant que les deux classes de composés fixent la même cible non parasitaire (soit l'hème), il serait possible de créer rationnellement une combinaison de composés quinoléine / diamidine. Ainsi, les souches résistantes à un des deux composés seraient plus sensibles à l'autre partenaire, retardant ainsi l'apparition de résistance. / In this thesis I have attempted to subject the issues of mode of action of recently synthesized bis cationic compounds and their possible interactions with chloroquine resistance. Antimalarial activities of representatives of various bis quarternary ammonium compounds, alkyl amidines (received from Dr.Vial group, Montpellier) and of bisbenzyl amidines (received from Chemistry group, Liverpool) activity have been investigated with chloroquine and pentamidine and looked for cross resistance with chloroquine. For this purpose two genetically modified allelically exchanged cell lines C3Dd2 and C2GCO3 modified on the chloroquine resistance-related PfCRT (P.falciparum chloroquine Resistance Transporter) gene were used. Among the benzyl amidines, a significant hypersensitivity tobis benzyl amidines was observed among the isolates bearing the mutant PfCRT. No such difference is observed for the bisalkyl amidines. To understand the mode of action and role of PfCRT, competitive binding assay to heme (which may mediate the well-known cellular accumulation of the compounds) and effect on heme crystallization assays (which is involved in the toxic effect against the intracellular parasite) were performed. All these compounds were shown to interact with heme in various degrees. Their activity was observed not to be correlating with heme crystallization inhibition. This is likely due to the structural differences between the compound which discriminate the compounds in the transport of the compound to the parasite and their mechanism of antimalarial activities. In addition I have studied the effect of pH on the pharmacological activity of the drugs using allelically exchanged genetically modified cell lines (for PfCRT) to characterize the importance of proton gradient on the transport of chloroquine and pentamidine to the intracellular parasite. Significant difference (reduced antimalarial activity with increased pH) in the activity of chloroquine was observed for both the strains. Despite of the high pKa values for pentamidine, there was significant difference in the sensitivity of the strains to this compound, when the pH is changed. As both the diamidines and choline analogs require transporters to cross the membrane barriers and enter the parasite where they accumulate I have also performed initial studies on the molecular characterization of a potential carrier in P.falciparum. Using basic bioinformatic tools, a gene encoding a P.falciparum protein with significant similarity to higher eukaryotes choline transporter was identified and preliminary work for its functional analysis was performed. In conclusion, this work establishes substantial differences between the various classes of bis-cationic compounds essentially (based on benzamidine and choline-analogs alkylkamidine series) concerning their interaction with the infected erythrocyte and their antimalarial activity. The series are diffentallly affected by the PfCRT mutation and the chloroquine resistance. Results suggest that it may be possible to use novel bisbenzyl amidines to specifically target quinoline resistant Plasmodium falciparum malaria, harbouring mutant pfcrt alleles. Taking this idea further and since both classes of compound target the same non-parasite target (heme), it may even be possible to rationally design a quinoline / diamidine drug combination, in which isolates resistant to one partner drug become more sensitive to the other partner, thus delaying the onset of resistance.