Dissertations / Theses on the topic 'Bass drum'

The main aim of this diploma thesis is to design the marketing mix of an alternative record label company that releases drum and bass music. In the theoretical part I try to describe the situation on the music market and outline how record labels work. I am specifying the tools of the marketing mix which are typical for a record label. In the practical part I'm pointing out the differences between the perception of mainstream and alternative music and I'm characterising the drum and bass scene. Based on this information I'm choosing the target audience and building the profile and the vision of my fictional

During the late 1980s and early 1990s, the United Kingdom's DJ-oriented electronic music community embraced new technologies and developed innovative techniques to master these technologies, resulting in the development of new genres. Three such genres, Hardcore, Jungle, and Drum & Bass (HJDB), emerged at the critical intersection between computer technology and the consumer market, which allowed computer music to be made through the use of home-based studios. The essential instrument in HJDB was the digital sampler, a device that offered musicians the ability to achieve realistic instrumentation through a "cut-and-paste" method of production. The thread that ties these three genres together was their unique usage of fast-paced sampled drums, derived primarily from breakbeats—samples of short percussion solos typically from 1960s to 1980s Funk and Jazz recordings. This dissertation explores a number of important issues that have not been addressed in prior writing on HJDB, and consists of three main objectives. The first is to provide a written history of the genres from the perspective of those that have made the music. This history catalogues the origins of the United Kingdom's DJ-oriented electronic music genres, the incorporation of breakbeats into this music that created the Hardcore genre, and developments that then resulted in the creation of the Jungle genre and subsequently the Drum & Bass genre. The second objective is to provide an explanation of the main technologies used in the creation of this music (e.g., the digital sampler) and the techniques developed by musicians to harness this technology. The third objective is to provide methods for the computational analysis of HJDB music, through automated determination of the breakbeats being used, detection of downbeat locations, and an estimation of the degree of rhythmic modification. Each of these objectives has been informed by over twenty interviews with musicians and label owners from throughout the history of HJDB. Computational methods based on HJDB-specific knowledge are shown to significantly outperform generalized music analysis techniques, highlighting the importance of style-specific approaches for computational musicology.<br>Durant la fin des années 80 et le début des années 90 au Royaume Uni, le monde de la musique électronique destinée aux DJ a adopté de nouvelles technologies et développé des techniques innovantes pour les maîtriser, aboutissant ainsi au développement de nouveaux genres. Trois de ces genres, Hardcore, Jungle, et Drum & Bass (HJDB), ont émergé au croisement critique entre la technologie numérique et le marché grand public, permettant la production de musique numérique dans les "home studios". L'instrument clé en HJDB était l'échantillonneur numérique, un appareil offrant aux musiciens la possibilité de réaliser une instrumentation réaliste grâce à une méthode de production "couper-coller". Le point commun entre ces trois genres était l'utilisation d'extraits de batterie au rythme rapide, principalement dérivés des "breakbeats"—courts extraits de solos de percussion typiques des enregistrements Funk et Jazz de 1960 à 1980.Ce travail explore un grand nombre de problématiques n'ayant pas été traitées dans la littérature HJDB et consiste en trois objectifs principaux. Le premier est de présenter l'histoire de ces genres musicaux du point de vue de leurs créateurs. Il s'agit de passer en revue les origines des différents genres de musique électronique destinée aux DJ—au Royaume-Uni, l'incorporation des "breakbeats" dans cette musique résultant dans la naissance du genre Hardcore, et les développements ayant mené au Jungle et par la suite au Drum & Bass. Le second objectif est de présenter les principales technologies utilisées dans la création de cette musique (par example, l'échantillonneur numérique) ainsi que les techniques développées par les musiciens pour maitriser ces technologies. Enfin, le troisième objectif est de fournir des méthodes pour l'analyse numérique de la musique HJDB, par la détection automatique des "breakbeats" utilisés, celle de la localisation des "downbeats", et une estimation du degrés de modification rythmique. Chacun de ces objectifs a été étayé par des entrevues faites avec plus que vingt musiciens et propriétaires de labels ayant participé à l'histoire du HJDB. Il a été démontré que les méthodes d'analyse informatique inspirées de la connaissance spécifique du HJDB surpassent significativement les techniques générales d'analyse musicale, mettant ainsi en évidence l'importance d'une approche spécifique du genre en musicologie numérique.

Studiens syfte var att skriva tre låtar inom EDM-genren med fokus på arrangering och mixning av baselement inom spannet 30–260 Hz. Metod: Med hjälp av olika analystekniker begrundades tre stycken referenslåtar i genrerna Drum and Bass, Glitch Hop och Progressiv house. Utifrån analyserna av dessa utformades tre stycken arrangeringsmodeller, vilka utgjorde grunden för mina tre produktioner. En jämförelse mellan referenslåtarna och produktionerna gjordes för att belysa skillnader och likheter. För att uppnå mina ideal av ett väl mixat basregister framkom det att det är av extra stor vikt att ta hänsyn till arrangering av baselement gentemot mixning av dessa. I mixstadiet visade det sig att verktyg såsom equalizer, kompressorer, expanders, distar och gaters m.m. hade stor betydelse för basregistrets kvalitet. Diskussion/slutsats: Då låtarna är i olika genrer och tempon kan riktlinjer underlätta mixstadiet såsom att snabba tempon inte är att föredra i kombination med en bastrumma med mycket subbas. Likaså att låtar med lägre tempo kan gynnas av att använda sidechain-kompression som ett rytmiskt verktyg. Att använda dynamiska verktyg såsom expanders kan skapa luft i mixen och motverka att den blir platt. Dist kan vara en bra metod för att ge basfrekvenser attityd och framhäva dem i mixen.

Thesis (Ph. D.)--University of California, San Diego, 2007.<br>Vita. Discusses the composer's use of the metaphors of space and perspective in his works, IN and Headspaces, the scores for which are included in the dissertation.

Identifying new potential treatment options for medical conditions that cause human disease burden is a central task of biomedical research. Since all candidate drugs cannot be tested with animal and clinical trials, in vitro approaches are first attempted to identify promising candidates. Likewise, identifying other essential relations (e.g., causation, prevention) between biomedical entities is also critical to understand biomedical processes. Hence, it is crucial to develop automated relation prediction systems that can yield plausible biomedical relations to expedite the discovery process. In this dissertation, we demonstrate three approaches to predict treatment relations between biomedical entities for the drug repositioning task using existing biomedical knowledge bases. Our approaches can be broadly labeled as link prediction or knowledge base completion in computer science literature. Specifically, first we investigate the predictive power of graph paths connecting entities in the publicly available biomedical knowledge base, SemMedDB (the entities and relations constitute a large knowledge graph as a whole). To that end, we build logistic regression models utilizing semantic graph pattern features extracted from the SemMedDB to predict treatment and causative relations in Unified Medical Language System (UMLS) Metathesaurus. Second, we study matrix and tensor factorization algorithms for predicting drug repositioning pairs in repoDB, a general purpose gold standard database of approved and failed drug–disease indications. The idea here is to predict repoDB pairs by approximating the given input matrix/tensor structure where the value of a cell represents the existence of a relation coming from SemMedDB and UMLS knowledge bases. The essential goal is to predict the test pairs that have a blank cell in the input matrix/tensor based on the shared biomedical context among existing non-blank cells. Our final approach involves graph convolutional neural networks where entities and relation types are embedded in a vector space involving neighborhood information. Basically, we minimize an objective function to guide our model to concept/relation embeddings such that distance scores for positive relation pairs are lower than those for the negative ones. Overall, our results demonstrate that recent link prediction methods applied to automatically curated, and hence imprecise, knowledge bases can nevertheless result in high accuracy drug candidate prediction with appropriate configuration of both the methods and datasets used.

The main theme of this thesis is that there is a common structural basis for drugs acting on the central nervous system (CNS), and that this concept may be used to design new CNS-active drugs which have greater specificity and hence less side-effects. To develop these ideas, the biological basis of how drugs modify CMS neurotransmission is described, and illustrated using dopaminergic pathways. An account is then given of the use of physicochemical concepts in contemporary drug design. The complete conformational analysis of several antipsychotic drugs is used to illustrate some of these techniques in the development of a model for antipsychotic drug action. After reviewing current structure-activity studies in several classes of CNS drugs (antipsychotics, anti-depressants, stimulants, hal1ucinogens, anticonvulsants and analgesics), a hypothesis for a common structural basis of CNS drug action is proposed- This is based on a topographical comparison of the X-ray structures of eight representative CNS-active drugs, and consists of three parts: 1.there is a common structural basis for the activity of many different CNS-active drug classes; 2. an aromatic ring and a nitrogen atom are the primary binding groups whose topographical arrangement is fundamental to the activity of these drug classes; 3. the nature and placement of secondary binding determines different classes of CNS drug activity. A four-Point model for this common structural basis is then defined using 14- CNS-active drug structures that include the original eight used in proposing the hypothesis. The coordinates of this model are: R1 (0. 3.5, 0), R2 (0, -3.5, O), N (4.8. -0.3, 1.4), and R3 (6.3, 1.3, 0), where R1 and R2 represent the point locations of a hydrophobic interaction of the common aromatic ring with a receptor, and R3 locates the receptor point for a hydrogen bond involving the common nitrogen, N. Extended structures were used to define the receptor points R1, R2 and R3, and the complete conformational space of each of the 14 molecules was considered. It is then shoun that the model may be used to predict whether a given structure is likely to show CNS activity: a search over 1,000 entries in the current Merck Index shows a high probability (82%) of CNS activity in compounds fitting the structural model. Analysis of CNS neurotransmitters and neuropeptides shows that these fit the common model well. Based on the available evidence supporting chemical evolution, protein evolution, and the evolution of neurotransmitter functions, it is surmised that the aromatic ring/nitrogen atom pharmacophore proposed in the common model supports the idea of the evolution of CNS receptors and their neurotransmitters, possibly from an aromatic amine or acety1cho1ine acting as a primaeval communicating molecule. The third point in the hypothesis trilogy is then addressed. The extensive conformation-activity analyses that have resulted in well-defined models for five separate CNS drug classes are used to map out the locations of secondary binding groups relative to the common model for anti-psychotics, antidepressants, analgesics, anticholinergics, and anticonvulsants. With this information, and knowledge derived from receptor-binding data, it is postulated that drugs having specified activity could be designed. In order to generate novel structures having a high probability of CNS-activity, a process of drug design is described in which known CNS structures are superimposed topographically using the common model as a template. Atoms regarded as superfluous may be selectively deleted and the required secondary binding groups added in predicted locations to give novel structures. It is concluded that this process provides the basis for the rational design of new lead compounds which could further be optimized for potent and specific CNS activity.

Quinolones are a clinically-useful class of antibacterial agents known to target DNA gyrase, a bacterial type II topoisomerase. Gyrase is unique among topoisomerases in its ability to introduce negative supercoils into DNA using the energy derived from ATP hydrolysis. The active enzyme is composed of two GyrA and two GyrB subunits, forming an A2B2 tetramer of molecular weight 374 kDa. The mechanism of supercoiling by gyrase involves the ATP-driven passage of one segment of DNA through a gyrase-stabilised double-stranded break in another. Tyrosine 122 of E. coli GyrA becomes covalently attached to DNA when gyrase breaks the phosphodiester bonds of DNA during supercoiling. When this residue is mutated to serine or phenylalanine, gyrase can no longer cleave or supercoil DNA, but can bind DNA normally. Rapid-gel filtration experiments have shown that quinolones can still bind to proteins bearing these mutations, suggesting that DNA cleavage by gyrase is not required for quinolone binding. Transcription by T7 and E. coli RNA polymerases is blocked by the presence of a gyrase-quinolone-DNA complex. Mapping of the transcription termination sites in the presence of gyrase and quinolones shows that blocking occurs about 10 to 20 base-pairs upstream of the gyrase cleavage site. Blocking of transcription by T7 RNA polymerase by a gyrase-quinolone complex on DNA does not occur when the active-site tyrosine of gyrase is mutated to serine, which indicates that the polymerase blocking requires DNA cleavage. Analysis of transcription in the absence of drug suggest that RNA polymerase does not displace gyrase from the template. DNA gyrase is also the target of the CcdB protein which is encoded by the F plasmid. When its action is not prevented by CcdA protein, CcdB is a potent cytotoxin. Using in vitro transcription by T7 RNA polymerase, it has been shown that CcdB complexed with gyrase can block transcription in a similar manner to the gyrase-quinolone complex. Furthermore, in the presence of CcdA, CcdB can no longer induce gyrase to block transcription.

Le contexte international de la pharmacoépidémiologie, marqué par la mise en œuvre d’un nombre croissant d’études multi-sources, a fait émerger un certain nombre de questionnements autour de la gestion de données conflictuelles ou de l’impact des choix méthodologiques sur les résultats.Accroître la confiance dans ces études observationnelles et renforcer leur crédibilité face aux données issues des essais cliniques représente un enjeu majeur, qui dépend étroitement de la robustesse des conclusions produites. Dans ce domaine, la mesure de l’exposition médicamenteuse revêt donc une importance toute particulière, tant pour des études portant sur l’estimation d’un risque ou d’un critère d’efficacité, que lors de la description des modalités d’utilisation en vie réelle. L’exposition médicamenteuse reste un phénomène complexe qui se caractérise la plupart du temps par des cycles discontinus, marqués par des évolutions de doses et la présence de médicaments concomitants. Compte tenu des caractéristiques pharmacodynamiques et pharmacocinétiques<br>The multinational context of pharmacoepidemiology, and the resulting increased number ofmulti-sources studies have generated concerns in relation with conflicting results and the question of the impact of methodological choices on study results. Increasing the confidence in the conclusions derived from these observational studies is a crucial issue, which is closely related to the robustness of the evidence produced. In this area, impact of drug exposure measurement and risk window might be crucial.Drug exposure is mostly characterized by discontinuous episodes, marked by changes in doses and presence of concomitant medications. Considering the pharmacokinetic and pharmacodynamics characteristics specific to each individual drug, the way in which the drug exposure is presented is of great importance. However, methods used for handling drug exposure episodes in electronic healthcare databases are varying widely according studies. However, the impact of these methods

Submitted by Oliveira Santos Dilzaná (dilznana@yahoo.com.br) on 2016-04-25T14:44:47Z No. of bitstreams: 1 DISSERTAÇÃO THIAGO NERI DA CONCEIÇÃO.pdf: 1639491 bytes, checksum: 25b3d865d476b42bed3a50c146ddc583 (MD5)<br>Approved for entry into archive by Ana Portela (anapoli@ufba.br) on 2016-04-29T19:38:47Z (GMT) No. of bitstreams: 1 DISSERTAÇÃO THIAGO NERI DA CONCEIÇÃO.pdf: 1639491 bytes, checksum: 25b3d865d476b42bed3a50c146ddc583 (MD5)<br>Made available in DSpace on 2016-04-29T19:38:47Z (GMT). No. of bitstreams: 1 DISSERTAÇÃO THIAGO NERI DA CONCEIÇÃO.pdf: 1639491 bytes, checksum: 25b3d865d476b42bed3a50c146ddc583 (MD5)<br>CNPq<br>Este trabalho toma por pretensão o tráfico de drogas no Nordeste, a tentativa de preenchimento de uma lacuna sociológica no que tange os estudos sobre crime/desvio no Brasil, através de um estudo de caso num bairro periférico da cidade de Salvador/BA. O Objetivo geral desta pesquisa é analisar as características do mercado de drogas na periferia e compreender seus impactos no cotidiano. Mediante a delimitação do objeto, utilizou-se técnicas qualitativas: observação participante, entrevistas e diário de campo, essas técnicas foram as que demonstraram maior eficácia diante do objetivo traçado. A amostra foi construída de modo gradativo, agrupando internamente: moradores, usuários de drogas e operadores do comércio de drogas. Narra-se aqui as diferentes fases assumidas pelo tráfico nesta localidade: fragmentada e concentrada. O primeiro marcado por uma violência difusa, o segundo por uma violência cirúrgica. Essas diferentes configurações são apresentadas de modo processual, destacando as características pertinentes a cada uma delas. Soma-se a isso, o reconhecimento de que essas fases têm caráter espiral; relaciona-se cada face assumida por esse mercado ilegal com a rotina da população onde o tráfico floresce, apontando a violência como instrumento que se expande para além do tráfico e se espraia às demais práticas cotidianas, não de modo epidêmico, mas como prática de controle. Concomitante a isso, aborda-se as práticas de segurança pública, que vão do policiamento de incursão à instalação de bases comunitárias de segurança, vislumbrando também cada uma dessas modalidades em suas interações com os moradores. This work has the intention drug trafficking in the Northeast, the attempt to fill a gap in sociological terms studies on crime / deviation in Brazil, through a case study in a suburb of the city of Salvador / BA. The general objective of this research is to analyze the drug market characteristics in the periphery and understand their impact on daily life. By defining the boundaries of the object, we used qualitative techniques: participant observation, interviews and field journal, these techniques were the ones that had the highest effectiveness on the stroke order. The sample was constructed a gradual way, gathering internally: residents, drug users and drug trade operators. This paper shows different phases assumed by trafficking: fragmented and concentrated, the first marked by widespread violence, the second by a surgically precise violence. These different settings are presented in a procedural way, highlighting the relevant characteristics for each of them. Added to this, there’s a recognition of the spiral character of these phases; each face assumed by illegal market is related with the routine of the population where drug trafficking flourishes, pointing violence as a tool that expands beyond the trafficking and reaches other daily practices. Concomitant to this, it approaches the public security practices, ranging from policing to the incursion and establishment of community security bases, also glimpsing each of these modalities in their interactions with the locals.

Il mio lavoro di tesi ha riguardato la preparazione di film per il wound healing a base di alginato e cheratina. Il primo passaggio ha previsto l'estrazione della cheratina da lana, proveniente da scarti tessili, e la sua successiva caratterizzazione mediante elettroforesi. In seconda battuta, è stato studiato il metodo più efficace per preparare un film stabile in soluzione acquosa di polisaccaride e proteina. In particolare, sono stati testati i seguenti meccanismi: attivazione dell'alginato per formazione di un estere succinimidilico, ossidazione dell'alginato con periodato di sodio, reazione di Maillard, crosslinking attraverso calcio cloruro e glutaraldeide. Quest'ultimo si è rivelato il metodo più opportuno da utilizzare. Quindi, si è proceduto con il caricamento dei due principi attivi (vitamina C e propoli). Quindi, i film ottenuti sono stati caratterizzati al fine di determinarne le principali proprietà. A questo proposito, sono stati studiati: lo swelling, la biodegradabilità ed i rilasci di acido ascorbico (utilizzando prima un metodo spettrofotometrico diretto e successivamente uno colorimetrico indiretto). Inoltre, tutti i film sono stati sottoposti ad analisi ATR, SEM e TGA. Sono in corso alcuni test biologici in vitro per verificare il ruolo della vitamina C sulla sintesi di nuovo collagene e per vedere l'effettiva attività antibatterica della propoli.

La chimie combinatoire et le screening à haut débit font l'objet de programmes de recherches ambitieux dans l'industrie pharmaceutique, en vue de repérer de nouvelles molécules actives. Mais le coût élevé associé à la synthèse et l'analyse d'un grand nombre de composés nécessitent l'apport d'outils informatiques capables d'optimiser la diversité moléculaire dans des grandes bases de données structurales et de présélectionner les molécules à tester. Dans ce contexte, appelé "Data Base Mining", la technique du "Self Organizing Maps" offre une visualisation attrayante de la distribution des composés d'une base de donnée sur une carte 2D, à partir de l'hyperespace défini par leurs descripteurs moléculaires. Par ailleurs, SOM, associé à une procédure de sélection de descripteur basée sur les algorithmes génétiques, nous a permis de discriminer plusieurs familles moléculaires d'une base de données pesticides. Cependant, SOM est une technique non supervisée qui implique l'utilisation d'outil complémentaire pour des études de prédiction. C'est la raison pour laquelle, des méthodes innovantes basées sur la logique floue, constituent un progrès fondamental pour des classifications complexes. Combinées à SOM ou utilisées directement dans l'hyperespace d'origine, ces techniques peuvent générer des modèles prédictifs reliant les descripteurs moléculaires aux propriétés biologiques. Des résultats très intéressants ont été obtenus dans la classification et la prédiction de composés actifs sur le système nerveux central. Toutes ces méthodes de classifications constituent une approche complémentaire indispensable aux techniques de modélisation moléculaires. A titre d'exemple, des études 3D QSAR ont été élaborées sur des inhibiteurs de la tyrosinase, enzyme intervenant dans la pigmentation. Bien que la structure cristallographique de l'enzyme ne soit pas disponible, ces procédures de modélisation ont permis d'établir des modèles QSAR robuste et de dériver un pharmacophore de la tyrosinase.

The Ketogenic diet (KD) is an alternative treatment option for people with drug-resistant epilepsy. It can reduce seizure frequency, but it is resource-intensive and may cause adverse side effects. Predictors of response – which, in the absence of specific metabolic disorders, are unknown – would improve patient selection and may enhance understanding of how the KD exerts its antiseizure effect. This project is concerned with identifying possible genetic markers of response to the KD. DNA was extracted from capillary blood taken from individuals who were following the KD for their epilepsy, or who had done so in the past. Individuals were classed as responders if they achieved ≥50 seizure reduction. Response was classified at various follow-up points, as well as a summary of response over time. Association studies were conducted using candidate gene (KCNJ11 and BAD) sequencing, genome-wide single nucleotide polymorphism array, and whole exome sequencing data to determine whether there was an over-representation of specific gene variants in KD responders, compared to non-responders. Common variation in KCNJ11 and BAD was not significantly associated with KD response. rs12204701 reached significance in the array-based genome-wide association study including common variants, with 3-month diet response as the phenotype. No significant results were obtained when summary diet response was taken as the phenotype. Using the gene-based c-alpha test with the exome sequencing data, including all exonic and splicing variants, ANKRD36C reached significance; using a pathway-based count of case-unique alleles test, the ‘ERBB1 Internalisation’ pathway reached significance. No further significant results were obtained from the exome sequencing data when using other gene- and pathway-based tests or when variants were further filtered according to predicted functional consequence. Other genes with large differences in responder/non-responder minor or alternative allele counts are also of interest. It is unknown how these may contribute to variability in KD response. Some common themes were identified amongst the genes and pathways of significance and suggestive significance: cell cycling, apoptosis, glucose homeostasis, neurological processes and triglyceride biosynthesis. It is biologically plausible that these processes influence KD response, although it is likely that many genes play a role. A larger sample size is needed in order to improve power to detect genotypic-phenotypic associations and increase confidence in the importance of the genes of interest.

Le développement des mémoires dynamiques (DRAM) à haute performance basées sur la structure métal-isolateur-métal (MIM) nécessite de remplacer la couche de dioxyde de silicium par des matériaux diélectriques à haute permittivité diélectrique. L'utilisation de ces isolants dits high-k permet de réduire la taille du dispositif DRAM tout en conservant une densité de capacité élevée et un faible courant de fuite pour diminuer la fréquence de rafraichissement. Parmi les nombreux matériaux high k, le dioxyde de titane (TiO2) est l'un des candidats les plus prometteurs en raison de sa constant diélectrique relativement élevée pouvant atteindre 170 dans le TiO2 cristallisé en phase rutile. De plus, il est possible d’obtenir cette phase à basse température par le procédé ALD (&lt; 250 °C) si le dépôt est réalisé sur un substrat RuO2 (phase rutile) grâce à une très faible différence de paramètres de maille entre les deux matériaux. L'objectif principal de cette thèse est d'étudier les mécanismes des réactions chimiques qui se produisant à l'interface RuO2/TiO2 lors du dépôt et leur influence sur les propriétés structurales et diélectriques du film TiO2, en particulier l'influence des espèces oxydantes, le plasma O2 et le H2O. L’influence des électrodes supérieure et inferieures sur les propriétés électriques et structurales de TiO2 a également été étudiée. Ensuite, la constante diélectrique, la conductivité ac et la tangente de perte des structures MIM à base d’oxyde de titane dopé aluminium ont été étudiés dans une gamme de fréquences large bande, de 1 Hz à 2 GHz. Enfin, la réalisation des MIM tridimensionnelles (3D) utilisant un substrat de silicium structuré en réseaux des trous coniques denses a été démontrée. Les structures MIM 3D réalisées ont permis d’augmenter sensiblement la densité de capacité tout en gardant de bonnes performances en termes de courant de fuite<br>The development of high performance dynamic random access memory (DRAM) based on metal-insulator-metal (MIM) structure made it necessary to replace the conventional silicon dioxide layer by dielectric materials with high dielectric constants. The use of these so-called high-k insulators allows aggressive scaling of DRAM devices while keeping high capacitance density and, more importantly, low leakage current. Among the numerous high k dielectrics, titanium dioxide (TiO2) is one of the most attractive candidate due to its rather high dielectric constant (k). Rutile TiO2 is the interesting phase due to its high dielectric constant and the possibility to deposit this phase at low temperature by ALD (&lt; 250 °C) by using RuO2 substrate thanks to a very small lattice mismatch between the two materials. The main objective of this thesis is to investigate the surface chemical reactions mechanisms at the RuO2/TiO2 interface and their influence on the ALD TiO2 film properties, especially the influence of oxidizing species, namely, H2O or O2 plasma. The influence of bottom and top electrode on electrical and structural proprieties of TiO2 MIM structure was also studied. Then, the dielectric constant, the ac conductivity and the loss tangent of aluminum doped titanium oxide are measured through a wide band frequency range, from 1 Hz to 2 GHz. Finally, the feasibility of three-dimensional (3D) MIM structures was studied by using dense array of truncated conical holes etched in a silicon substrate. The 3D MIM capacitors showed a large increase in the capacitance density while retaining very good electrical properties especially a leakage current comparable to planar MIM devices

Ce travail de thèse s'inscrit dans le cadre d'une collaboration entre le LHFA à Toulouse et le département CMC (Chemistry Manufacturing and Control) de Sanofi à Vitry-sur-Seine, et a pour but de développer de nouveaux nanovecteurs à base de conjugués polymériques biodégradables et biocompatibles capables d'être utilisés pour la vectorisation de principe actif. Dans le premier chapitre, une étude bibliographique a été détaillée afin de comprendre les principes de la vectorisation et de faire le point sur les principales avancées réalisées pour la vectorisation d'agents thérapeutiques par administration intraveineuse, en particulier à l'aide de polymères biodégradables et biocompatibles, tels que le PLA et le PEG. Dans le second chapitre, tirant profit de la catalyse organique de la ROP, de nouveaux conjugués macromoléculaires à base de PEG-PLA/Taxane, avec une architecture particulière de type " Y " positionnant le PA entre les blocs de PLA et de PEG, ont été synthétisés. Des conjugués de type PEG-PLA-Taxane linéaires ont aussi été préparés pour comparer l'influence de l'architecture sur les propriétés des nanovecteurs. La composition chimique de ces nanoconjugués a été précisément caractérisée à l'aide de plusieurs techniques analytiques (RMN, DOSY, UPLC, SEC. . . ). Dans le troisième chapitre, les conjugués macromoléculaires ont été formulés dans un milieu aqueux pour donner naissance à des nanoparticules de conformation " cœur-couronne ". Les deux différentes architectures permettent d'obtenir des NPs stables. Ensuite, L'utilisation de la spectroscopie RMN dans le D2O a permis de localiser le PA au sein des NPs : à l'interface hydrophile/hydrophobe pour les conjugués " Y " et dans le centre hydrophobe pour les conjugués linéaires. Enfin, les études de libération du PA in vitro ont montré des profils prolongés et différents en fonction de l'architecture et de la nature de l'espaceur : la libération la plus rapide a été obtenue avec la structure " Y " et avec l'espaceur " diglycolique "<br>This thesis is a collaboration between the LHFA and the CMC department (Chemistry Manufacturing and Control) of Sanofi at Vitry-sur-Seine, and aims to develop new nanocarriers based on biodegradable and biocompatible polymeric conjugates that could be used for the vectorization of active ingredient. In the first chapter, a bibliographic review is detailed to understand the principles of vectorization and to review the main achievements for the delivery of therapeutic agents by intravenous administration, particularly with biodegradable and biocompatible polymers, such as PLA and PEG. In the second chapter, benefiting from the ROP organic catalysis, new macromolecular conjugates based on PEG-PLA/Taxane are synthesized, with a particular architecture (Y-shape), positioning the PA between PLA and PEG blocks. Linear conjugates of PEG-PLA-taxane were also prepared to evaluate the influence of the architecture on the nanocarriers properties. The chemical composition of these nanoconjugates was precisely characterized using several analytical techniques (NMR, DOSY, UPLC, SEC. . . ). In the third chapter, the macromolecular conjugates were formulated in an aqueous medium to obtain nanoparticles with a "core-shell" conformation. The two different architectures allow to obtain stable NPs. Then, the API was localized into NPs using the NMR spectroscopy in D2O : API at the hydrophilic/hydrophobic interface for the Y-shape conjugates and in the center of the hydrophobic core for the linear conjugates. Finally, in vitro release of the API showed sustained and different profiles depending on the shape and on the nature of the spacer : the fastest release was obtained with the Y-shape structure and the diglycolic spacer

The human immunodeficiency virus type 1 (HIV-1) protease (PR) is a critical drug target as it is responsible for virion maturation. Mutations within the active site (1°) of the PR directly interfere with inhibitor binding while mutations distal to the active site (2°) to restore enzymatic fitness. Increasing mutation number is not directly proportional to the severity of resistance, suggesting that resistance is not simply additive but that it is interdependent. The interdependency of both primary and secondary mutations to drive protease inhibitor (PI) resistance is grossly understudied. To structurally and dynamically characterize the direct role of secondary mutations in drug resistance, I selected a panel of single-site mutant protease crystal structures complexed with the PI darunavir (DRV). From these studies, I developed a network hypothesis that explains how mutations outside the active site are able to perpetuate changes to the active site of the protease to disrupt inhibitor binding. I then expanded the panel to include highly mutated multi-drug resistant variants. To elucidate the interdependency between primary and secondary mutations I used statistical and machine-learning techniques to determine which specific mutations underlie the perturbations of key inter-molecular interactions. From these studies, I have determined that mutations distal to the active site are able to perturb the global PR hydrogen bonding patterns, while primary and secondary mutations cooperatively perturb hydrophobic contacts between the PR and DRV. Discerning and exploiting the mechanisms that underlie drug resistance in viral targets could proactively ameliorate both current treatment and inhibitor design for HIV-1 targets.

The human immunodeficiency virus type 1 (HIV-1) protease (PR) is a critical drug target as it is responsible for virion maturation. Mutations within the active site (1°) of the PR directly interfere with inhibitor binding while mutations distal to the active site (2°) to restore enzymatic fitness. Increasing mutation number is not directly proportional to the severity of resistance, suggesting that resistance is not simply additive but that it is interdependent. The interdependency of both primary and secondary mutations to drive protease inhibitor (PI) resistance is grossly understudied. To structurally and dynamically characterize the direct role of secondary mutations in drug resistance, I selected a panel of single-site mutant protease crystal structures complexed with the PI darunavir (DRV). From these studies, I developed a network hypothesis that explains how mutations outside the active site are able to perpetuate changes to the active site of the protease to disrupt inhibitor binding. I then expanded the panel to include highly mutated multi-drug resistant variants. To elucidate the interdependency between primary and secondary mutations I used statistical and machine-learning techniques to determine which specific mutations underlie the perturbations of key inter-molecular interactions. From these studies, I have determined that mutations distal to the active site are able to perturb the global PR hydrogen bonding patterns, while primary and secondary mutations cooperatively perturb hydrophobic contacts between the PR and DRV. Discerning and exploiting the mechanisms that underlie drug resistance in viral targets could proactively ameliorate both current treatment and inhibitor design for HIV-1 targets.

5-Bromo-2'-deoxyuridine (BrdU), a thymidine analogue, is genotoxic and teratogenic. The exposure of mouse embryos to BrdU at doses that cause malformations induces oxidative stress and an embryonic stress response characterized by an increase in c-Fos dependent AP-1 DNA binding. The goal of this thesis was to test the hypothesis that development is disturbed at sites where BrdU is incorporated into DNA, triggering oxidative stress and c-Fos induction. Gestation day 9 CD-1 mice were treated with BrdU and embryos were obtained for immunolocalization of BrdU, 8-oxoguanine, a biomarker for oxidative stress, and c-Fos. BrdU incorporation into DNA was dispersed throughout the embryo. In contrast, the staining for 8-oxoguanine and c-Fos were highest in the neuroepithelium. BrdU incorporation was not affected by the pre-administration of N-acetyl-cysteine (NAC), an anti-oxidant, although both 8-oxoguanine and c-Fos staining were decreased. Thus, the response of the embryo to insult is tissue specific.

Os hidróxidos duplos lamelares (HDLs) são matrizes inorgânicas bidimensionalmente estruturadas que apresentam propriedades diversas e interessantes para a formação de materiais híbridos. Em especial na área medicinal e farmacológica, o emprego de HDLs com lamelas contendo os íons Mg/Al e Zn/Al tem mostrado resultados promissores como antiácidos e como carregadores de fármacos. Por outro lado, anti-inflamatórios não esteroidais como o sulindaco são moléculas orgânicas ionizáveis que podem ser intercalados em HDLs levando à formação de materiais híbridos com potencial para uso na área farmacêutica. Este trabalho consistiu na síntese e na caracterização de materiais híbridos baseados em matrizes de HDLs de ZnRAl e MgRAl (R é a razão molar MII/Al) e o ânion derivado do fármaco sulindaco (abreviados M2IIAl-Sul). Foram avaliadas as condições experimentais mais apropriadas para a obtenção de materiais com alta cristalinidade e alto conteúdo do fármaco. Os materiais híbridos preparados por um método simples de coprecipitação (one pot) à temperatura ambiente e a 55°C apresentaram capacidade de carregamento de 50 a 55% (em massa) de sulindaco. Os difratogramas de raios X dos híbridos M2IIAl-Sul indicaram a formação de materiais mais cristalinos a 55°C, com várias reflexões basais indicando alto grau de organização das lamelas e distância interplanar de cerca de 2,74 nm. O arranjo bidimensional interdigitado dos ânions sulindaco entre as lamelas foi proposto através do mapa de densidade eletrônica unidimensional (1D Plot). A análise morfológica por microscopias eletrônicas de varredura e transmissão revelou a formação de estruturas nanométricas planas, flexíveis e desordenadas no caso do Mg2Al-Sul; as partículas de Zn2Al-Sul também apresentam nanoscrolls, com cerca de 100 nm de diâmetro, uma morfologia que não havia sido reportada até então para HDLs. Os espectros vibracionais no infravermelho dos materiais híbridos mostraram bandas típicas do sulindaco atribuídas ao vS=O em 1011 e 1039 cm-1, ao vC-F em 1163 cm-1 e bandas referentes aos estiramentos antissimétrico e simétrico do grupo COO- em aproximadamente 1564 e 1394 cm-1, que confirmam a presença do sulindaco na forma aniônica no sistema híbrido. Deslocamentos de picos observados nos espectros de ressonância magnética nuclear de carbono-13 no estado sólido sugerem que o confinamento entre as lamelas promove alterações na densidade de carga da espécie orgânica. As técnicas espectroscópicas confirmam a manutenção da integridade química do fármaco após o processo de intercalação. As atribuições dos espectros vibracionais e de ressonância magnética nuclear foram efetuadas com o auxílio do método da Teoria do Funcional de Densidade (DFT) para a espécie orgânica. A avaliação do comportamento térmico por termogravimetria acoplada à calorimetria exploratória diferencial e à espectrometria de massas (TG-DTA-MS) evidencia sensível aumento na estabilidade térmica do fármaco intercalado em matriz de Zn2Al. Esse fato foi interpretado através da análise dos espectros vibracionais de amostras aquecidas a 250°C. Propõe-se que a desidroxilação das lamelas do HDL de zinco promove a coordenação do grupo carboxilato do ânion sulindaco ao metal divalente, aumentando sua estabilidade térmica. Os ensaios de dissolução in vitro evidenciam um mecanismo difusional de liberação modificada (prolongada) do fármaco a partir do material híbrido, que ocorre em cerca do dobro do tempo observado para o sulindaco não intercalado. Resultados preliminares de estudo in vivo de pastilhas dos materiais híbridos implantadas em região intramuscular de ratos atestam a biocompatibilidade e a antigenicidade dos materiais HDL-Cl e dos híbridos HDL-Sul.<br>Layered double hydroxides (LDHs) are two-dimensionally structured inorganic matrices that show diverse and interesting properties for hybrid materials development. Particularly in the medical and pharmaceutical area, the use of LDHs with layers containing Zn/Al and Mg/Al ions has shown promising results. On the other hand, nonsteroidal drugs such as sulindac are ionizable organic molecules that can be intercalated into LDH leading to the formation of hybrid materials with potential usage in the pharmaceutical area. This work involved the synthesis and characterization of hybrid materials based on matrices of LDHs ZnRAl and MgRAl (R is the MII/Al molar ratio) and the anion derived from the sulindac drug (abbreviated M2IIAl-Sul). The most suitable experimental conditions for obtaining materials with high crystallinity and also high drug contents were evaluated. Hybrid materials prepared by a simple coprecipitation method (one pot) at room temperature and 55 °C showed loading capacity of about 50 - 55% (by mass) of sulindac. X-ray diffraction revealed the formation of higher crystalline materials at 55 °C; various basal reflections indicate a high degree of layers stacking and an interlayer distance of about 2.74 nm to M2IIAl-Sul hybrids. A two-dimensional array of interdigitated sulindac anions between the layers was proposed by one-dimensional electron density map (1D Plot). The morphological analysis by electronic scanning and transmission microscopies revealed the formation of flat, flexible and disordered nanoscale structures in the case of Mg2Al-Sul; nanoscrolled particles of about 100 nm in diameter are also present in Zn2Al-Sul sample, a morphology which had not been reported so far for LDHs. The vibrational spectra of hybrid materials in the infrared showed the characteristic sulindac bands assigned to vS=O at 1011 and 1039 cm-1, the vCF at 1163 cm-1 and bands related to the antisymmetric and symmetric stretching of the -COO- group at approximately 1564 and 1394 cm-1 which confirmed the presence of the anion sulindac in the hybrid system. Shift peaks observed in carbon-13 nuclear magnetic resonance spectra in solid state suggest that the confinement between the layers promotes variations in the organic species charge density. Spectroscopic techniques confirm the maintenance of the chemical integrity of the drug after the intercalation process. The spectral assignments of the organic species were supported by Density Functional Theory (DFT). The evaluation of the thermal behavior by thermogravimetry coupled with differential scanning calorimetry and mass spectrometry (TG-DSC-MS) shows substantial increase in the thermal stability of the drug intercalated into Zn2Al matrix. This effect was interpreted by analysis of vibrational spectra of samples calcined at 250 °C. It is proposed that dehydroxylation of the Zn2Al layers promotes the coordination of the sulindac carboxylate group to the divalent metal. In vitro dissolution tests of hybrid material show a diffusional mechanism of modified drug release (sustained), which occurs in about twice the time observed for sulindac not intercalated. Preliminary results from in vivo study of hybrid materials\' pellets implanted in the intramuscular region of rats attest to biocompatibility and antigenicity of LDH-Cl and LDH-Sul hybrid materials

Las enfermedades y otros procesos biológicos son complejos y están producidos por la interacción de múltiples factores genéticos y ambientales. En las últimas décadas, los métodos tradicionales reduccionistas han dado paso a estrategias holísticas que proporcionan una visión más amplia del problema. En concreto, la biología de sistemas permite el análisis tanto de las partes como de las interacciones que formas los organismos biológicos y sus funciones. Además, el desarrollo de las tecnologías de alto rendimiento y de herramientas bioinformáticas facilitan la compresión de complejos procesos biológicos. A lo largo de esta tesis, utilizamos métodos propios de la biología de sistemas para aportar nuevos conocimientos relevantes para resolver diversas cuestiones biológicas. Para ello integramos múltiple datos biológicos para comprender las bases moleculares que conducen al desarrollo y progresión de enfermedades complejas y de la respuesta a determinados tratamientos farmacológicos.<br>Diseases and other biological processes are complex, involving the interplay of multiple genetic and environmental determinants. In the last decades, traditional reductionist approaches have given way to holistic strategies that provide a broader view. In particular, systems biology allows for the examination not only of the parts but also of the interactions that assemble biological organisms and their functions. Moreover, the development of high-throughput technologies and bioinformatics tools facilitate the understanding of complex biological processes. In this thesis, we benefit from systems biology methods to provide relevant insight into various biological questions. We integrate multiple biological data to understanding the molecular bases that lead to the development and progression of complex diseases as well as the treatment responses.

Work presented in this thesis is in two parts. Part one: The X-ray crystal structures of potential antimicrobial drug targets. The protein IspF (2C-methyl-D-erythritol-2,4-cyclodiphosphate synthase, EC: 4.6.1.12) from two pathogens (Burkholderia cenocepacia and Plasmodium falciparum) has been investigated. IspF is an enzyme of isoprenoid-precursor biosynthesis and is considered to be a potential drug target. The results of structural and fragment-screening efforts presented here inform early stage drug discovery efforts. The structure of the PabC protein (4-amino-4-deoxychorismate lyase, EC: 4.1.3.38) from the Gram-negative pathogen Pseudomonas aeruginosa was also determined. PabC is involved in the production of para-aminobenzoic acid on the path to folate. Comparisons with previously solved PabC structures identified a spatially conserved tyrosine residue in the active site and suggest that a re-evaluation of a published mechanism is warranted. Part two: Immunity proteins in the Gram-negative Type VI secretion system. The X-ray crystal structures of the proteins Rap1a and Rap2a from Serratia marcescens, inhibitors of the peptidoglycan amidase toxins secreted by some Gram-negative bacteria employing the Type VI secretion pathway, were determined by molecular replacement and analysed.

[ES] La cristalografía de rayos X es una potente técnica para la resolución de la estructura atómica de macromoléculas. La información generada, tiene gran impacto sobre diferentes campos relacionados con la investigación básica y aplicada, como son la biomedicina y diseño de fármacos, al igual que en el desarrollo de aplicaciones nanotecnológicas y biotecnológicas. Esta Tesis se centra en determinadas problemáticas actuales y en las proteínas involucradas en las mismas (TryR, eEF1A2 y CBDP35), siendo éstas sujeto de desarrollo biotecnológico en los campos de la biomedicina, farmacia y de la industria alimentaria, en el que la cristalografía de rayos X juega un papel crucial para dilucidar sus estructuras atómicas y funciones. En consideración a la biomedicina y diseño de fármacos, hemos resuelto la estructura de la Tripanotión reductasa (TryR) de Leishmania infantum en complejo con potentes inhibidores de su actividad oxidorreductasa, con potencial de desarrollo como fármacos. Así, se ha caracterizado la unión y mecanismo de acción de éstos inhibidores. TryR es una reconocida diana farmacológica para el tratamiento de la enfermedad de Chagas, la Tripanosomiasis Humana Africana y la leishmaniosis, ya que desempeña un papel crucial y esencial en el metabolismo redox de los parásitos de la familia Trypanosomatidae. Además, se han analizado los parámetros de cristalización y difracción de novedosos inhibidores de la dimerización de TryR, cuyo diseño racional se basa en la unión a la interfaz de dimerización de la misma. La oncoproteína eEF1A2, involucrada en múltiples funciones celulares y sujeto de numerosas modificaciones post-traduccionales, se une al fármaco anticancerígeno plitidepsina. La cristalografía de rayos X, combinada con experimentos de espectrometría de masas, se han utilizado como herramientas para identificar nuevas modificaciones post-traduccionales y características estructurales en eEF1A2:GDP. Una modificación única, la adición de etanolamina fosfoglicerol (EPG) a aminoácidos conservados (Glu301 y Glu374 en mamíferos), se ha observado aquí por primera vez. El análisis estructural de estos hallazgos facilita la comprensión de las múltiples funciones y regulaciones de eEF1A2. La adquisición de una muestra conformacionalmente homogénea de eEF1A2:GTP, necesaria para la unión a la plitidepsina, ha sido evaluada en ensayos de cristalización del complejo terciario de eEF1A2: GTP: plitidepsina. Con respecto al dominio de unión a la pared celular de la endolisina PlyP35 codificada por el fago P35 de Listeria monocytogenes (CBDP35), hemos resuelto la estructura cristalina de CBDP35 en un complejo con ácido teicoico natural de L. monocytogenes serovar 1/2a. Esta estructura es el primer módulo de unión a la pared celular en complejo con ácidos teicoicos jamás dilucidado. El análisis estructural reveló los principales determinantes para la unión de la pared celular bacteriana, en particular, el mecanismo molecular del reconocimiento de N-acetil-d-glucosamina, una decoración de carácter glicosídico en ácidos teicoicos de serovares patógenos de L. monocytogenes. Estos hallazgos arrojan luz sobre el desarrollo biotecnológico de nuevas herramientas en la industria alimentaria y las terapias derivadas de fagos para detectar y tratar infecciones bacterianas.<br>[CAT] La cristal·lografia de raig X és una potent tècnica per a la resolució de l'estructura atòmica de macromolècules. La informació generada té gran impacte sobre diferents camps relacionats amb la investigació bàsica i aplicada, com són la biomedicina i disseny de fàrmacs, igual que en el desenvolupament d'aplicacions nanotecnológiques i biotecnològiques. Aquesta Tesi es centra en determinades problemàtiques actuals i en les proteïnes involucrades en les mateixes (TryR, eEF1A2 i CBDP35), sent estes subjecte de desenvolupament biotecnològic en els camps de la biomedicina, farmàcia i de la indústria alimentària, en el que la cristal·lografia de raig X juga un paper crucial per a dilucidar les seues estructures atòmiques i funcions. En consideració a la biomedicina i disseny de fàrmacs, hem resolt l'estructura de la Tripanotión reductasa (TryR) de Leishmania infantum en complex amb potents inhibidors de la seua activitat oxidorreductasa, amb potencial de desenrotllament com a fàrmacs. Així, s'ha caracteritzat la unió i mecanisme d'acció d'estos inhibidors. TryR és una reconeguda diana farmacològica per al tractament de la malaltia de Chagas, la Tripanosomiasi Humana Africana i la leishmaniosi, ja que exerceix un paper crucial i essencial en el metabolisme redox dels paràsits de la família Trypanosomatidae. A més, s'han analitzat els paràmetres de cristal·lització i difracció de nous inhibidors de la dimerizació de TryR, el disseny racional dels quals es basa en la unió a la interfície de dimerización de la mateixa. L'oncoproteína eEF1A2, involucrada en múltiples funcions cel·lulars i subjecte de nombroses modificacions posttraduccionals, s'unieix al fàrmac anticancerigen plitidepsina. La cristal·lografia de raig X, combinada amb experiments d'espectrometria de masses, s'han utilitzat com a ferramentes per a identificar noves modificacions posttraduccionals i característiques estructurals en eEF1A2:GDP. Una modificació única, l'addició d'etanolamina fosfoglicerol (EPG) a aminoàcids conservats (Glu301 i Glu374 en mamífers), s'ha observat ací per primera vegada. L'anàlisi estructural d'estes troballes facilita la comprensió de les múltiples funcions i regulacions d'eEF1A2. L'adquisició d'una mostra conformacionalmente homogènia d'eEF1A2:GTP, necessària per a la unió a la plitidepsina, ha sigut avaluada en assajos de cristal·lització del complex terciari d'eEF1A2: GTP: plitidepsina. Respecte al domini d'unió a la paret cel·lular de l'endolisina PlyP35 codificada pel fago P35 de Listeria monocytogenes (CBDP35), hem resolt l'estructura cristal·lina de CBDP35 en un complex amb àcid teicoico natural de L. monocytogenes serovar 1/2a. Esta estructura és el primer mòdul d'unió a la paret cel·lular en complex amb àcids teicoicos mai dilucidat. L'anàlisi estructural va revelar els principals determinants per a la unió de la paret cel·lular bacteriana, en particular, el mecanisme molecular del reconeixement de N-acetil-d-glucosamina, una decoració de caràcter glicosídico en àcids teicoicos de serovares patògens de L. monocytogenes. Estes troballes fan llum sobre el desenrotllament biotecnològic de noves ferramentes en la indústria alimentària i les teràpies derivades de fagos per a detectar i tractar infeccions bacterianes.<br>[EN] X-ray crystallography is a powerful technique for atomic structure resolution of macromolecules. The information generated impacts different fields involving basic and applied research on biomedicine and drug design and the development of nanotechnology and biotechnological applications. This dissertation focuses on current problematics and the target proteins involved (TryR, eEF1A2 and CBDP35) that are in sight for biotechnological development in the biomedical, pharmaceutical and food industry fields, in which X-ray crystallography plays a crucial role in the elucidation of their atomic structures and functions. Attaining to biomedical and drug design problematics, we have solved the structure of Leishmania infantum TryR in complex with potent oxidoreductase inhibitors prone to further development as anti-trypanosomal drugs, thereby characterizing their binding and mechanism of action. This protein is a long recognized drug target for the treatment of Chagas disease, Human African Trypanosomiasis and leishmaniasis, as it plays a crucial and essential role in the redox-metabolism of the Trypanosomatidae parasites. Moreover, the crystallization and diffraction parameters of novel TryR dimerization disruptors have been assayed for inhibitors which have been rationally designed to bind the dimerization interface of TryR. The "moonlighting" oncoprotein eEF1A2 is known to be highly post-translationally modified and to bind the anticancer drug plitidepsin. X-ray crystallography, combined with mass-spectrometry experiments, have been used as tools to identify novel post-translational modifications and structural features in eEF1A2:GDP. A unique modification, namely the addition of ethanolamine phosphoglycerol (EPG) to conserved glutamic residues (Glu301 and Glu374 in mammals), has been here observed for the first time. Structural analysis of these findings facilitate the understanding of eEF1A2's multiple functions and regulations. The acquirement of a conformationally homogenous eEF1A2:GTP sample, necessary for plitidepsin binding, has been has been assayed for eEF1A2:GTP:plitidepsin complex crystallization. Regarding the cell wall binding domain of Listeria monocytogenes phage-encoded endolysin PlyP35 (CBDP35), we have solved the crystal structure of CBDP35 in complex with natural Listeria serovar 1/2a teichoic acid. This structure is the first cell wall binding module in complex with teichoic acids ever elucidated. Structural analysis revealed the main determinants for bacterial cell-wall binding, in particular, the molecular mechanism of N-acetyl-d-glucosamine recognition, a glycosidic moiety in teichoic acids of pathogenic serovars of L. monocytogenes. These findings shed light upon the biotechnological development of new tools in the food industry and phage-derived therapies to detect and treat bacterial infections.<br>Agradecer al Ministerio de Educación, Cultura y Deporte por haberme proporcionado el contrato FPU (FPU14/03190) que me ha permitido desarrollar esta Tesis Doctoral en el Instituto de Química-Física “Rocasolano” del Consejo Superior de Investigaciones Científicas (IQFR-CSIC), así como la financiación otorgada para poder realizar mi estancia predoctoral en el laboratorio del Prof. Hammershmidt, en Greifswald, Alemania (EST17/00751).<br>Carriles Linares, AÁ. (2019). STRUCTURAL BIOMEDICINE: CHARACTERIZATION OF THE STRUCTURAL BASIS IN PROTEIN-DRUG RECOGNITION IN DIFFERENT HUMAN DISEASES [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/130844<br>TESIS

Submitted by Nuzia Santos (nuzia@cpqrr.fiocruz.br) on 2016-10-17T11:27:29Z No. of bitstreams: 1 Tese_SC_MarianaMartinsGonzagadoNascimento.pdf: 218266 bytes, checksum: 09bb50de7e46f75bfeee14bd0625a861 (MD5)<br>Approved for entry into archive by Nuzia Santos (nuzia@cpqrr.fiocruz.br) on 2016-10-18T13:10:31Z (GMT) No. of bitstreams: 1 Tese_SC_MarianaMartinsGonzagadoNascimento.pdf: 218266 bytes, checksum: 09bb50de7e46f75bfeee14bd0625a861 (MD5)<br>Made available in DSpace on 2016-10-18T13:10:31Z (GMT). No. of bitstreams: 1 Tese_SC_MarianaMartinsGonzagadoNascimento.pdf: 218266 bytes, checksum: 09bb50de7e46f75bfeee14bd0625a861 (MD5) Previous issue date: 2016<br>Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.<br>O uso de medicamentos potencialmente inadequados (MPI) para idosos pode estar associado a mais riscos que benefícios e sua utilização tem sido documentada internacionalmente. Nesta perspectiva, os objetivos deste trabalho foram: (1) numa abordagem transversal, estimar a prevalência de utilização de MPI entre idosos residentes na Região Metropolitana de Belo Horizonte (RMBH), Minas Gerais, e os fatores associados a essa prática; (2) longitudinalmente, investigar se o uso de MPI constitui fator de risco independente para a mortalidade entre idosos residentes em comunidade. A abordagem transversal foi baseada nos dados coletados junto a uma amostra representativa da população idosa com 60 anos ou mais residentes na RMBH (n=1.158); no cumprimento do segundo objetivo, utilizou-se os dados coletados junto à coorte idosa do Projeto Bambuí (n=1.586), composta em 1997 e acompanhada, anualmente, até 2011. Para definição do uso de MPI, variável dependente no estudo transversal e exposição de interesse no estudo longitudinal, utilizou-se o critério de Beers 2012. Variáveis sociodemográficas, de condições de saúde, de utilização de serviços de saúde e número de medicamentos foram utilizadas em caráter exploratório no estudo dos fatores associados ao uso de MPI (etapa transversal) e como variáveis de ajuste na investigação da associação entre uso de MPI e mortalidade (vertente longitudinal). A análise da prevalência e dos fatores associados ao uso de MPI foi baseada no modelo de regressão de Poisson; a investigação da associação do uso de MPI e mortalidade foi realizada por meio do modelo dos riscos proporcionais de Cox, adotando-se em ambos os casos, o nível de significância estatística de 5%. Na RMBH, a prevalência do uso de MPI foi de 43,3%. O sexo feminino, o número de doenças crônicas e a polifarmácia apresentaram-se positiva e independentemente associadas ao uso de MPI, sendo a última a variável mais fortemente associada. Em Bambuí, o uso de MPI mostrou-se como fator de risco para mortalidade entre os idosos da coorte. Nossos resultados apontam para a necessidade da seleção de alternativas terapêuticas mais seguras para idosos.<br>Potentially inappropriate medications (PIMs) for the elderly can be associated with greater risks than benefits and its use has been reported internationally. Having this into consideration, the objectives of this study were: (1) in a cross-sectional study, to estimate the prevalence of PIMs use among elderly residents of the Metropolitan Region of Belo Horizonte (MRBH), Minas Gerais, and the associated factors; (2) in a longitudinal study, to investigate if the PIM use constitutes a risk factor for mortality among community dwelling elderly. The cross-sectional approach was based on data from a representative sample of the elderly population (60 years or older) living in the MRBH (n=1.158). To fulfill the second objective, data from Bambuí elderly cohort (composed in 1997 and followed annually until 2011) study was used. Beers criteria (2012) were used to define PIM use, which was the dependent variable in the crosssectional study and exposition factor in the longitudinal study. Socio-demographic variables, health status, healthcare services use and number of medications were used as exploratory variables in the study involving associated factors (crosssectional study), and as adjustment variables in the investigation of association between PIM use and mortality (longitudinal study). Prevalence analysis and associated factors were performed using Poisson regression model. To investigate the association between PIMs use and mortality, Cox proportional hazards model was used. A 0.05 significance level was adopted for all analyzes. The prevalence of PIM use was 43.3% in the MRBH. Female gender, number of chronic conditions and polypharmacy were positively and independently associated with PIM use, the latter having been the most strongly associated factor. In Bambuí, PIM use was identified as a risk factor for mortality among the elderly in the cohort. These results indicate the need for selection of safer therapeutic alternatives for elderly patients.

Thesis (M.S. in Defense Analysis)--Naval Postgraduate School, December 2006.<br>Thesis Advisor(s): Kalev Sepp. "December 2006." AD-A462 564. Includes bibliographical references (p.83-89). Also available via the World Wide Web.

The Major Facilitator Superfamily groups a vast number of secondary transporters that import or export distinct substrates. Among these, multidrug antiporters constitute a peculiar class of transporters, both because of their multispecificity, recognizing structurally very diverse substrates, and because of their transport mechanism, that relies on bilayer-mediated extrusion of cytotoxic compounds. An accurate and detailed description of the conformational changes that underlie the transport cycle is still lacking and the structural basis for energetic coupling in these transporters has not been elucidated, with so far only limited crystallographic evidence available. We investigate the molecular basis of secondary multidrug transport with biochemical and biophysical studies on LmrP, a Major Facilitator Superfamily multidrug transporter from Lactococcus lactis. We used extensive continuous-wave electron paramagnetic resonance and double electron-electron resonance measurements on a library of spin-labeled LmrP mutants to uncover the conformational states involved in transport and to investigate how protons and ligands shift the equilibrium between conformers to enable transport. We find that the transporter switches between outward-open and outward-closed conformations depending on the protonation states of specific acidic residues forming a transmembrane protonation relay. We observe that substrate binding restricts the conformational freedom of LmrP and induces localized conformational changes. Our data allows to build a model of secondary multidrug transport wherein substrate binding initiates the transport cycle by opening the extracellular side to protons. Subsequent protonation of membrane-embedded acidic residues induces substrate release to the extracellular side and triggers a cascade of conformational changes that culminates in a proton release to the intracellular side. Parallel to this, we have optimized our purification and expression protocol in order to set up crystallization trials on LmrP. Through extensive screening and optimization of the lipidation state of LmrP, using ad hoc methods for sample preparation, we were able to obtain low-resolution diffracting crystals. By improving our lipidation technique and modifying the lipid composition we further improved crystal quality. Other factors such as ligand addition, the presence of secondary detergent and additives for controlling phase separation and nucleation were tested, paving the way to high resolution structure determination of LmrP.<br>Doctorat en Sciences<br>info:eu-repo/semantics/nonPublished

ABC (ATP-binding cassette) transporters are involved in translocate a wide spectrum of molecules across the lipid bilayer and some of them are associated with various diseases. They also have an important role in multidrug resistance (MDR) in cancer, which has been a major obstacle in cancer chemotherapy and in the treatment of leishmaniasis. While diverse transporters may confer MDR phenotype in bacteria, in human it is mainly achieved by five ABC proteins, among them P-glycoprotein/(ABCB1). To understand the structural basis of P-gp inhibitory activity, to determine the ligand binding sites within P-gp and to guide the design of more potent P-gp inhibitors, we performed i) a 3D-QSAR model using CoMSIA on a set of sesquiterpenes, ii) molecular docking simulations of various compounds in a homology model of LMDR1, a P-gp-like transporter belonging to the Leishmania ABC family and iii) and a complete study of sesquiterpenes interaction with human P-gp.<br>Los transportadores ABC (ATP binding cassette), encargados de transportar un amplio espectro de moléculas a través de la bicapa lipídica, pueden estar asociados con diversas enfermedades. Juegan un papel importante en la multirresistencia a fármacos (MDR), obstáculo importante en la quimioterapia del cáncer y en el tratamiento de leishmaniasis. En bacterias varios transportadores pueden conferir el fenotipo MDR, en humanos son principalmente cinco, entre ellos la glicoproteína P/(ABCB1). Para comprender la base estructural de la actividad inhibidora de P-gp, determinar los sitios de unión de los ligandos a P-gp y diseñar inhibidores de P-gp más potentes, se realizó i) un modelo 3D-QSAR usando CoMSIA en un conjunto de sesquiterpenos, ii) simulaciones de acoplamiento molecular de varios compuestos en un modelo de homología de LMDR1, transportador de la P-gp perteneciente a la familia ABC de Leishmania y iii) un estudio completo de interacción entre sesquiterpenos y la P-gp humana.

La polymédication est définie comme l’administration de nombreux médicaments de façon simultanée ou comme l’administration d’un nombre excessif de médicaments. L’objectif général de la thèse était de décrire la polymédication chronique et le risque associé à partir des bases de données médico-administratives. Un indicateur de description de la polymédication chronique, défini par l’usage concomitant et chronique de 5 médicaments ou plus, a été développé pour décrire la prévalence et les médicaments impliqués. À partir des données de l’Assurance Maladie Française, nous avons estimé que la polymédication chronique concernait plus de 5 % des français de tout âge, et impliquait majoritairement les médicaments cardiovasculaires. Afin d’approfondir la caractérisation de la polymédication chronique, une description de la part potentiellement inappropriée a été conduite en utilisant les données de l’Assurance Maladie Française et de la Veterans Health Administration aux États-Unis. À partir de ces données nous avons estimé que la part inappropriée de la polymédication chronique était majeure puisque concernant, en France, 65 % des sujets âgés (≥ 65 ans) et 46 % des sujets d’âge moyen (45-64 ans), et aux États-Unis, 67 % des vétérans. Les inhibiteurs de la pompe à proton, les médicaments psychiatriques et les médicaments du diabète étaient les médicaments potentiellement inappropriés les plus fréquents. La polymédication, qu’elle soit appropriée ou non est un facteur de risque de mortalité. Chez les vétérans américains nous avons montré que la polymédication chronique et les médicaments potentiellement inappropriés augmentaient le risque de décès<br>Polypharmacy is defined as the administration of many drugs at the same time or as the administration of an excessive number of drugs. The main objective of this thesis was to describe chronic polypharmacy and the associated risk, from medico-administrative databases. A specific indicator of chronic polypharmacy, defined by five or more concomitant and chronic drug uses, was developed to describe the prevalence and the involved drugs. Based on data from the National French Health Insurance, we estimated that chronic polypharmacy concerned more than 5% of all aged French individuals, and involved mainly cardiovascular drugs. To further describe chronic polypharmacy, a description of inappropriateness was carried out using data from the National French Health Insurance and the United States Veterans Health Administration. We estimated that the inappropriateness of chronic polypharmacy was major. It concerned 65% of French older adults (≥ 65 years old), 46% of French middle-aged adults (45-64 years old) in France, and 67% of American veterans. Proton pump inhibitors, psychiatric drugs and diabetes drugs were the most common potentially inappropriate medications. Furthermore, we have shown that chronic polypharmacy, whether appropriate or not, is associated with increased mortality. In American veterans, chronic polypharmacy and potentially inappropriate medications both increased the risk of all-cause death

La confirmation d’une hypersensibilité médicamenteuse est importante car la plupart des cas allégués ne sont pas confirmés. Le diagnostic repose sur l’interrogatoire et le bilan allergologique, ce dernier comprenant surtout des tests in vivo. Ces tests ne sont pas dénués de risque. Ce travail se propose de répondre avec une méthodologie originale (pour le domaine de l’hypersensibilité médicamenteuse), à des questions en suspens visant la nécessité, les modalités et l’utilité du bilan allergologique, en prenant principalement mais pas exclusivement le modèle des hypersensibilités aux ß-lactamines. J’ai utilisé plusieurs approches méthodologiques, appliquées à une large base de données d’hypersensibilités médicamenteuses. Dans un premier temps, j’ai exploité les données cliniques rétrospectives afin de construire 2 modèles de diagnostic d’hypersensibilité aux ß-lactamines et j’ai ensuite testé leurs performances diagnostiques sur un échantillon prospectif de patients. Les modèles atteignent globalement une sensibilité de 50%, ce qui est difficilement acceptable, dans un contexte iatrogénique. Secondairement, j’ai réalisé le passage des protocoles de tests de provocation aux ß-lactamines, d’une étape avec paliers purement empiriques, à un protocole basé sur des données issues d’une analyse de survie. Autres 3 articles ont suivi une méthodologie similaire : les patients ayant eu un bilan allergologique négatif pour un médicament donné ont été re-contactés et interrogés au moyen d’un questionnaire. Le service rendu au patient a été calculé par le taux de patients ayant repris (sans réaction) le médicament autorisé (plus de 90% pour 3 classes médicamenteuses analysées)<br>Most alleged cases of hypersensitivity reactions following drug administration are actually ruled out by drug allergy work up. The diagnosis is based on clinical history and allergy tests, mainly in vivo tests. These tests carry a considerable risk of iatrogeny. The purpose of this thesis was to address some unmet needs regarding the need, technical aspects and utility of the drug allergy work up, using an original methodology applied to the drug hypersensitivity field. It focuses mainly (but not only) on drug hypersensitivity reactions to ß-lactams. I used different statistical methods, applied to a large database, the Drug Allergy and Hypersensitivity Database. First (Article 1), I used retrospective clinical data to build 2 models for ß-lactam hypersensitivity diagnosis. I then tested these models on a prospective sample, in order to analyze their diagnostic performances. The overall sensitivity of the 2 models is around 50%, which is unacceptable in an iatrogenic context. Second (Article 2), I worked on empirical protocols of drug provocation tests and I identified steps for data-driven, evidence-based protocols by means of survival analysis. The Articles 3, 4 and 5 were conceived following a similar methodology: patients with a negative drug allergy work-up for a certain drug were called and questioned on whether they had been exposed to this same drug, following allergy tests. High negative predictive values of these tests, with more than 90% of patients tolerating subsequent administration without any hypersensitivity reaction, were obtained for 3 different drug classes

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico<br>NanopartÃculas à base de goma de cajueiro foram obtidas por duas rotas. Na primeira rota, nanopartÃculas de goma de cajueiro foram sintetizadas atravÃs da reaÃÃo de enxertia com poliacrilamida (goma de cajueiro-g-poliacrilamida). Na segunda, a goma foi modificada pela reaÃÃo do polissacarÃdeo com anidrido propiÃnico. A formaÃÃo das nanopartÃculas atravÃs do derivado do polissacarÃdeo foi feita por auto-estruturaÃÃo atravÃs da aplicaÃÃo do processo de diÃlise. NanopartÃculas de goma de cajueiro-g-poliacrilamida foram caracterizadas por espectroscopia na regiÃo do infravermelho (IV), anÃlise termogravimÃtrica (TGA) e tamanho de partÃcula. A espectroscopia no IV confirma a enxertia de poliacrilamida na estrutura do polissacarÃdeo e indica que parte da poliacrilamida foi hidrolisada. A curva TGA da goma de cajueiro-g-poliacrilamida mostra 5 eventos de degradaÃÃo. Este modelo à muito diferente das curvas TGA da goma de cajueiro e da poliacrilamida, no qual foram observados apenas dois eventos de degradaÃÃo, confirmando a modificaÃÃo da estrutura da goma de cajueiro. A mudanÃa da razÃo goma/acrilamida/agente reticulante nÃo interfere no tamanho de partÃcula. NanopartÃculas com distribuiÃÃo unimodal e mÃdia de tamanho de 8 nm foram obtidas. As partÃculas tÃm a superfÃcie carregada negativamente, inferido pelo potencial zeta negativo observado. As nanopartÃculas mostraram propriedades sensÃveis ao pH. AgregaÃÃo foi observada apÃs a liofilizaÃÃo da suspensÃo de nanopartÃculas. Cloroquina, um fÃrmaco usado no tratamento da malÃria, foi incorporado Ãs nanopartÃculas. A liberaÃÃo do fÃrmaco revelou-se dependente do pH, sendo observado liberaÃÃo controlada durante 8 dias em pH 6,0. ModificaÃÃo da goma de cajueiro com anidrido propiÃnico foi conformado por espectroscopia na regiÃo do IV, ressonÃncia magnÃtica nuclear (RMN) de 13C e TGA. A estimativa do grau de substituiÃÃo (GS) de grupos propionato na goma de cajueiro foi calculado por RMN 13C e encontrou-se igual a 1,7. A formaÃÃo de nanopartÃculas atravÃs de diÃlise foi feita usando acetona ou DMSO como solvente para nanopartÃculas e Ãgua como nÃo-solvente. DistribuiÃÃo de tamanho de partÃcula unimodal foi observado em ambos os solventes. Tamanho de partÃcula entre 42 e 142 nm foi observado, dependendo da concentraÃÃo da soluÃÃo ou do solvente usado no processo de diÃlise. Menores partÃculas foram obtidas usando DMSO como solvente. NanopartÃculas mostraram-se estÃveis por um perÃodo de tempo de 120 dias. AgregaÃÃo foi observada depois da liofilizaÃÃo da suspensÃo de nanopartÃculas.<br>Nanoparticles based on cashew gum were obtained by two rotes. In the first rote, nanoparticles of cashew gum were synthesized by a graft reaction with polyacrylamide (cashew gum-g-polyacrylamide), in the second one the gum was modified by reaction of the polysaccharide with propionic anhydride. The formation of the nanoparticle from the propionic polysaccharide derivative was made by selfassembly applying the dialysis process. Nanoparticles from the cashew gum-graftacrylamide were characterized by infrared spectroscopy (FT-IR), thermalgravimetric analysis (TGA) and particle size. Infrared spectroscopy confirms the graft of acrylamide onto polysaccharide structure and indicates that part of polyacrylamide has been hydrolyzed. TGA analysis of cashew gum-g-polyacrylamide shows five degradation events. This pattern is very different of the TGA curves from cashew gum and polyacrylamide where only two degradation events were observed, confirming the modification on the cashew gum structure. The change in gum/acrylamide/crosslinking agent ratio does not interfere on particle size. Nanoparticles with unimodal particle size distribution and average particle size of 8 nm were obtained. The particles have a negative surface; this was inferred by the negative zeta potential observed. The nanoparticle shows pH sensitive properties. Aggregation was observed after freeze-drying of nanoparticle suspension. Chloroquine a drug for malaria treatment was incorporated into the nanoparticles. The release of the drug shows to be pH dependent with a controlled released observed for 8 days been observed at pH 6.0. Modification of cashew gum with propionic anhydride was confirmed by FT-IR and 13C- nuclear magnetic resonance(NMR) spectroscopy and TGA analysis. A estimative of degree of substitution (DS) of propionate groups on cashew gum was calculate by 13C-NMR and found to be equal to 1.7. The formation of nanoparticle through dialysis process was made using acetone or DMSO as solvent for nanoparticles and H2O as a non-solvent. Unimodal particle size distribution was observed in both solvents. Particle size ranging from 42 to 142 nm were obtained their size were found to depend on solution concentration or solvent used in the dialysis process. Small particles were obtained using DMSO as solvent. Nanoparticles were shown to be stable for a storage time up to 120 days. Aggregation was also observed after freeze-drying of nanoparticle suspension.

L’objectif de ce travail était d’analyser de manière conjointe différents indicateurs d’abus, de dépendance et de détournement de médicaments psychoactifs en conditions réelles d’utilisation issus de sources de données hétérogènes afin d’en présenter une vision synthétique. Les sources de données utilisées dans ce travail sont issues des outils et programmes des Centres d’Evaluation et d’Information sur la Pharmacodépendance – Addictovigilance (CEIP-A). Elles permettent de mesurer directement l’abus auprès de populations spécifiques de patients dépendants ou sous traitement de substitution par l’enquête OPPIDUM (Observation des Produits Psychotropes Illicites ou Détournés de leur Utilisation Médicamenteuse). Ces outils permettent aussi de mesurer le détournement via la mesure du nombre d’ordonnances falsifiées par l’enquête OSIAP (Ordonnances Suspectes, Indicateur d’Abus Possible) ainsi que la mesure du phénomène de « Doctor Shopping » (chevauchement d’ordonnances) et la mesure du nombre de patients présentant des comportements déviants (issu d’une analyse de classification) à partir de bases de données de remboursement de l’assurance maladie. Cette approche multi-sources a d’abord été appliquée à l’analyse de l’importance du détournement d’usage du clonazépam (Publication n°1). Ce travail a permis de mettre en évidence le détournement émergent du clonazépam et a surtout permis d’illustrer les difficultés à faire émerger cette information de manière cohérente et standardisée au travers des différentes sources de données. Ensuite, l’un des pré-requis d’un système de surveillance de l’abus et du détournement de médicaments étant de permettre d’étudier des tendances évolutives, nous avons proposé la méthode de classification visant à établir des profils de sujets déviants de manière à analyser l’évolution de détournement d’usage de méthyphénidate sur quatre années (Publication n°2). Cette méthode de classification a par la suite, été appliquée de manière conjointe à une méthode de mesure du « Doctor Shopping » pour étudier le détournement d’usage de la Buprenorphine Haut Dosage (BHD) dans la région PACA-Corse (Publication n°3). Cette étude nous a non seulement permis de mettre en évidence un problème important de détournement d’usage de la BHD mais elle nous a permis aussi de montrer la concordance entre ces deux méthodes (mesure du « Doctor Shopping » et méthode de classification) et d’évaluer leurs apports respectifs pour la surveillance de l’abus des médicaments. Ces deux méthodes ont par la suite été analysées de manière conjointe aux données issues des enquêtes OPPPIDUM et OSIAP pour permettre d’étudier et de comparer le détournement d’usage des médicaments de la famille des benzodiazépines (Publication n°4) et des opioïdes (Publication n°5). Cette approche multi-sources permet de limiter les biais inhérents à chaque méthode ou source prise isolément. L’ensemble de nos travaux met en exergue la pertinence d’un tel système pour évaluer l’abus d’un médicament mais aussi pour le comparer à d’autres substances. Néanmoins, le développement d’un tel système appliqué au domaine de la pharmacodépendance est relativement nouveau, et nécessite des améliorations tant dans l’intégration d’autres sources de données, que dans la méthodologie employée pour intégrer et synthétiser l’information ainsi obtenue. Finalement, cette thèse a montré que les CEIP-A avaient le potentiel pour mettre en œuvre un système multi-sources pouvant apporter une réelle contribution à l’étude de la pharmacodépendance en France<br>The objective of this work was to analyze abuse, dependence and diversion of psychoactive medicines in real settings using jointly different indicators issued from mixed datasources in order to present a synthetic vision. The datasources used in this work are issued from the tools developed by the Centres for Evaluation and Information on Pharmacodependency (CEIP). They allow to measure directly drug abuse with specific populations of dependent patients or under opiate treatment (OPPIDUM (Observation of the Illicit Psychotropic Products or Diverted from their Medicinal Use) survey)). These tools also allow to measure the diversion via the measure of the phenomenon of “doctor shopping” (overlapping of prescriptions) and the measure of the number of patients presenting a deviant behaviour from general health insurance databases; then they measure diversion through falsified prescriptions presented at pharmacies (the OSIAP (Forged prescriptions indicating potential abuse) survey).This multisources approach has been firstly applied to analyse abuse and diversion of clonazepam (1st publication). This study has highlighted the emerging problem of diversion of clonazepam, after flunitrazepam and has also illustrated the difficulty of analysing with consistency the information gathered by these different datasources. A good system for controlling drug diversion and abuse has to allow analysing trends. We have so proposed a classification method aiming at revealing profile of subjects with deviant behaviour to use it on an evolutive manner so as to study diversion of methylphenidate on a four year period (2nd publication). This classification method has then been applied jointly with a method measuring the “doctor shopping” to analyse diversion of High Dosage Buprenorphine (HDB) (3rd publication). This study has revealed an important problem of diversion of HDB, has also demonstrated that the two methods were globally concordant and has allowed to evaluate their advantages for the controlling of the abuse and diversion of prescription drugs. These two last methods have then been analysed jointly with data from the OPPIDUM and OSIAP surveys to allow to study and compare diversion of benzodiazepine drugs (4th publication) and opioids drugs (5th publication). This multisource approach allows to limit biases linked to each method seen individually. Our work points out the relevance of such a multisources system to estimate the abuse of a prescription drug and to compare it with the other substances. Nevertheless, the development of such a system applied to the domain of the drug dependency is relatively new, and requires improvements concerning the integration of the other sources of data and the methodology used to join and synthetize the information obtained. Finally, such a system "multi-sources” has the potential to exist and to make a real contribution to the domain of the drug dependency in France