Avalia����o de fiabilidade de sistemas el��tricos e de automa����o em instala����es de frio industrial

Disserta����o para a obten����o do grau de Mestre em Engenharia Electrot��cnica Ramo de Energia/Automa����o e Eletr��nica Industrial

Predictors of consistent condom use among Portuguese women attending family planning clinics

Women account for 30% of all AIDS cases reported to the Health Ministry in Portugal and most infections are acquired through unprotected heterosexual sex with infected partners. This study analyzed socio-demographic and psychosocial predictors of consistent condom use and the role of education as a moderator variable among Portuguese women attending family planning clinics. A cross-sectional study using interviewer-administered fully structured questionnaires was conducted among 767 sexually active women (ages 18���65). Logistic regression analyses were used to explore the association between consistent condom use and the predictor variables. Overall, 78.7% of the women were inconsistent condom users. The results showed that consistent condom use was predicted by marital status (being not married), having greater perceptions of condom negotiation self-efficacy, having preparatory safer sexual behaviors, and not using condoms only when practicing abstinence. Living with a partner and having lack of risk perception significantly predicted inconsistent condom use. Less educated women were less likely to use condoms even when they perceive being at risk. The full model explained 53% of the variance in consistent condom use. This study emphasizes the need for implementing effective prevention interventions in this population showing the importance of taking education into consideration.

Melanosome transfer between melanocytes and keratinocytes

RESUMO: A pele �� o maior ��rg��o do corpo humano e a sua pigmenta����o �� essencial para a sua colora����o e prote����o contra os efeitos nocivos da radia����o ultravioleta (UV). A pigmenta����o da pele resulta essencialmente de tr��s processos: a s��ntese e o armazenamento de melanina pelos melan��citos, em organelos especializados denominados melanossomas; o transporte dos melanossomas dentro dos melan��citos; e finalmente, a transfer��ncia dos melanossomas para os queratin��citos adjacentes. Nos queratin��citos, a melanina migra para a regi��o perinuclear apical da c��lula para formar um escudo protetor,respons��vel pela prote����o do DNA dos danos causados pela radia����o UV. Os melan��citos est��o localizados na camada basal da epiderme e contactam com 30-40 queratin��citos. Em conjunto, estas c��lulas formam a ���unidade melano-epid��rmica���. Apesar dos processos de s��ntese e transporte de melanina nos melan��citos estarem bastante bem caracterizados, os mecanismos moleculares subjacentes �� transfer��ncia inter-celular de melanina s��o menos conhecidos e ainda controversos. Dados preliminares obtidos pelo nosso grupo, que se basearam na observa����o de amostras de pele humana por microscopia electr��nica, indicam que a forma predominante de transfer��ncia de melanina na epiderme consiste na exocitose dos melanossomas pelos melan��citos e subsequente endocitose da melanina por queratin��citos. Para al��m disso sabe-se que as prote��nas Rab, que controlam o tr��fego membranar, est��o envolvidas em v��rias etapas de pigmenta����o da pele, nomeadamente na biog��nese e no transporte de melanina. Assim, dado o seu papel fundamental nestes processos, question��mo-nos sobre o seu envolvimento na transfer��ncia de melanina. Com este trabalho, propomo-nos a expandir o conhecimento atual sobre a transfer��ncia de melanina na pele, atrav��s do estudo detalhado dos seus mecanismos moleculares, identificando as prote��nas Rab que regulam o processo. Pretendemos tamb��m confirmar o modelo de exo/endocitose como sendo o mecanismo principal de transfer��ncia de melanina. Primeiro, explor��mos a regula����o da secre����o de melanina pelos melan��citos e analis��mos o papel de prote��nas Rab neste processo. Os resultados foram obtidos recorrendo a um m��todo in vitro, desenvolvido previamente no laborat��rio, que avalia a quantidade de melanina segregada para o meio de cultura por espectrofotometria, e ainda por microscopia, contando o n��mero de melanossomas transferidos para os queratin��citos. Atrav��s de co-culturas de melan��citos e queratin��citos, verificou-se que os queratin��citos estimulam a liberta����o de melanina dos melan��citos para o meio extra-celular, bem como a sua transfer��ncia para os queratin��citos. Al��m disso, a prote��na Rab11b foi identificada como um regulador da exocitose de melanina e da sua transfer��ncia para os queratin��citos. De facto, a diminui����o da express��o de Rab11b em melan��citos provocou a redu����o da secre����o de melanina estimulada por queratin��citos, bem como da transfer��ncia desta. Em segundo lugar, para complementar o nosso estudo, centr��mos a nossa investiga����o na internaliza����o de melanina por queratin��citos. Especificamente, usando uma biblioteca de siRNA, explor��mos o envolvimento de prote��nas Rab na capta����o de melanina por queratin��citos. Como primeira abordagem, us��mos esferas fluorescentes como substituto de melanina, avaliando os resultados por citometria de fluxo. No entanto, este m��todo revelou-se ineficaz uma vez que a internaliza����o destas esferas �� independente do recetor PAR-2 (recetor 2 ativado por protease), que foi previamente descrito como essencial na capta����o de melanina por queratin��citos Posteriormente, foi desenvolvido um novo protocolo de endocitose baseado em microscopia, usando melanossomas sem a membrana envolvente (melanocores) purificados do meio de cultura de melan��citos, incluindo um programa inform��tico especialmente desenhado para realizar uma an��lise semi-automatizada. Ap��s internaliza����o, os melanocores acumulam-se na regi��o perinuclear dos queratin��citos, em estruturas que se assemelham ao escudo supranuclear observado na pele humana. Seguidamente, o envolvimento do recetor PAR-2 na capta����o de melanocores por queratin��citos foi confirmado, utilizando o novo protocolo de endocitose desenvolvido. Para al��m disso, a necessidade de quatro prote��nas Rab foi identificada na internaliza����o de melanocores por queratin��citos. A redu����o da express��o de Rab1a ou Rab5b em queratin��citos diminuiu significativamente o n��vel de internaliza����o de melanocores, enquanto o silenciamento da express��o de Rab2a ou Rab14 aumentou a quantidade de melanocores internalizados por estas c��lulas. Em conclus��o, os resultados apresentados corroboram as observa����es anteriores, obtidas em amostras de pele humana, e sugerem que o mecanismo de transfer��ncia predominante �� a exocitose de melanina pelos melan��citos, induzida por queratin��citos, seguida por endocitose pelos queratin��citos. A pigmenta����o da pele tem implica����es tanto ao n��vel da cosm��tica, como ao n��vel m��dico, relacionadas com foto-envelhecimento e com doen��as pigmentares. Assim sendo, ao esclarecer quais os mecanismos moleculares que regulam a transfer��ncia de melanina na pele, este trabalho pode conduzir ao desenvolvimento de novas estrat��gias para modular a pigmenta����o da pele.----------------ABSTRACT: Skin pigmentation is achieved through the highly regulated production of the pigment melanin in specialized organelles, termed melanosomes within melanocytes. These are transported from their site of synthesis to the melanocyte periphery before being transferred to keratinocytes where melanin forms a supra-nuclear cap to protect the DNA from UVinduced damage. Together, melanocytes and keratinocytes form a functional complex, termed ���epidermal-melanin unit���, that confers color and photoprotective properties to the skin. Skin pigmentation requires three processes: the biogenesis of melanin; its intracelular transport within the melanocyte to the cell periphery; and the melanin transfer to keratinocytes. The first two processes have been extensively characterized. However, despite significant advances that have been made over the past few years, the mechanisms underlying inter-cellular transfer of pigment from melanocytes to keratinocytes remain controversial.Preliminary studies from our group using electron microscopy and human skin samples found evidence for a mechanism of coupled exocytosis-endocytosis. Rab GTPases are master regulators of intracellular trafficking and have already been implicated in several steps of skin pigmentation. Thus, we proposed to explore and characterize the molecular mechanisms of melanin transfer and the role of Rab GTPases in this process. Moreover, we investigated whether the exo/endocytosis model is the main mechanism of melanin transfer. We first focused on melanin exocytosis by melanocytes. Then, we started to investigate the key regulatory Rab proteins involved in this step by establishing an in vitro tissue culture model of melanin secretion. Using co-cultures of melanocytes and keratinocytes, we found that keratinocytes stimulate melanin release and transfer. Moreover, depletion of Rab11b decreases keratinocyte-induced melanin exocytosis by melanocytes. In order to determine whether melanin exocytosis is a predominant mechanism of melanin transfer, the amount of melanin transferred to keratinocytes was then assayed in conditions where melanin exocytosis was inhibited. Indeed, Rab11b depletion resulted in a significant decrease in melanin uptake by keratinocytes. Taken together, these observations suggest that Rab11b mediates melanosome exocytosis from melanocytes and transfer to keratinocytes. To complement and extend our study, we of melanin by keratinocytes. Thus, we aimed to explore the effect of depleting Rab GTPases on melanin uptake and trafficking within keratinocytes. As a first approach, we used fluorescent microspheres as a melanin surrogate. However, the uptake of microspheres was observed to be independent of PAR-2, a receptor that is required for melanin uptakecentred our attention in the internalization of melanin by keratinocytes. Thus, we aimed to explore the effect of depleting Rab GTPases on melanin uptake and trafficking within keratinocytes. As a first approach, we used fluorescent microspheres as a melanin surrogate. However, the uptake of microspheres was observed to be independent of PAR-2, a receptor that is required for melanin uptake.Therefore, we concluded that microspheres were uptaken by keratinocytes through a different pathway than melanin. Subsequently, we developed a microscopy-based endocytosis assay using purified melanocores (melanosomes lacking the limiting membrane) from melanocytes, including a program to perform a semi-automated analysis. Melanocores are taken up by keratinocytes and accumulate in structures in the perinuclear area that resemble the physiological supranuclear cap observed in human skin. We then confirmed the involvement of PAR-2 receptor in the uptake of melanocores by keratinocytes, using the newly developed assay. Furthermore, we identified the role of four Rab GTPases on the uptake of melanocores by keratinocytes. Depletion of Rab1a and Rab5b from keratinocytes significantly reduced the uptake of melanocores, whereas Rab2a, and Rab14 silencing increased the amount the melanocores internalized by XB2 keratinocytes. In conclusion, we present evidence supporting keratinocyte-inducedmelanosome exocytosis from melanocytes, followed by endocytosis of the melanin core by keratinocytes as the predominant mechanism of melanin transfer in skin. Although advances have been made, there is a need for more effective and safer therapies directed at pigmentation disorders and also treatments for cosmetic applications. Hence, the understanding of the above mechanisms of skin pigmentation will lead to a greater appreciation of the molecular machinery underlying human skin pigmentation and could interest the pharmaceutical and cosmetic industries.

Fungal communities in archives: assessment strategies and impact on paper conservation and human health

The main results presented in this PhD Dissertation have been published in interna-tional journals included in the Science Citation Index (SCI)

Portuguese banks and the economic adjustment programme: balance sheet adjustments and impact on profitability

Double Degree. A Work Project presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA ��� School of Business and Economics and a Masters Degree in Finance from Louvain School of Management

Light harvesting in solar cells using natural pigments from red fruits adsorbed to mesoporous TiO2

Nature has developed strategies to present us with a wide variety of colours, from the green of leaves to the bright colours seen in flowers. Anthocyanins are between these natural pigments that are responsible for the great diversity of colours seen in flowers and fruits. Anthocyanins have been used to sensitize titanium dioxide (TiO2) in Dye-Sensitized Solar Cells (DSSCs). DSSCs have become one of the most popular research topic in photovoltaic cells due to their low production costs when compared to other alternatives. DSSCs are inspired in what happens in nature during photosynthesis. A primary charge separation is achieved by means of a photoexcited dye capable of performing the electron injection into the conduction band of a wide band-gap semiconductor, usually TiO2. With this work we aimed to synthesize a novel mesoporous TiO2 structure as the semiconductor in order to increase the dye loading. We used natural occurring dyes such as anthocyanins and their synthetic flavylium relatives, as an alternative to the widely used metal complexes of Ru(II) which are expensive and are environmentally unsafe. This offers not only the chance to use safer dyes for DSSCs, but also to take profit of waste biological products, such as wine and olive oil production residues that are heavily loaded with anthocyanin dyes. We also performed a photodegradation study using TiO2 as the catalyst to degrade dye contaminants, such as those from the wine production waste, by photo-irradiation of the system in the visible region of the light spectrum. We were able to succeed in the synthesis of mesoporous TiO2 both powder and thin film, with a high capacity to load a large amount of dye. We proved the concept of photodegradation using TiO2 as catalyst. And finally, we show that wine production waste is a possible dye source to DSSCs application.

Bioluminescent sensor proteins for point-of-care therapeutic drug monitoring.

For many drugs, finding the balance between efficacy and toxicity requires monitoring their concentrations in the patient's blood. Quantifying drug levels at the bedside or at home would have advantages in terms of therapeutic outcome and convenience, but current techniques require the setting of a diagnostic laboratory. We have developed semisynthetic bioluminescent sensors that permit precise measurements of drug concentrations in patient samples by spotting minimal volumes on paper and recording the signal using a simple point-and-shoot camera. Our sensors have a modular design consisting of a protein-based and a synthetic part and can be engineered to selectively recognize a wide range of drugs, including immunosuppressants, antiepileptics, anticancer agents and antiarrhythmics. This low-cost point-of-care method could make therapies safer, increase the convenience of doctors and patients and make therapeutic drug monitoring available in regions with poor infrastructure.

The clinical and biological impact of new pathogen inactivation technologies on platelet concentrates.

Since 1990, several techniques have been developed to photochemically inactivate pathogens in platelet concentrates, potentially leading to safer transfusion therapy. The three most common methods are amotosalen/UVA (INTERCEPT Blood System), riboflavin/UVA-UVB (MIRASOL PRT), and UVC (Theraflex-UV). We review the biology of pathogen inactivation methods, present their efficacy in reducing pathogens, discuss their impact on the functional aspects of treated platelets, and review clinical studies showing the clinical efficiency of the pathogen inactivation methods and their possible toxicity.

Substitution of (R,S)-methadone by (R)-methadone: Impact on QTc interval.

Methadone is administered as a chiral mixture of (R,S)-methadone. The opioid effect is mainly mediated by (R)-methadone, whereas (S)-methadone blocks the human ether-��-go-go-related gene (hERG) voltage-gated potassium channel more potently, which can cause drug-induced long QT syndrome, leading to potentially lethal ventricular tachyarrhythmias. To investigate whether substitution of (R,S)-methadone by (R)-methadone could reduce the corrected QT (QTc) interval, (R,S)-methadone was replaced by (R)-methadone (half-dose) in 39 opioid-dependent patients receiving maintenance treatment for 14 days. (R)-methadone was then replaced by the initial dose of (R,S)-methadone for 14 days (n = 29). Trough (R)-methadone and (S)-methadone plasma levels and electrocardiogram measurements were taken. The Fridericia-corrected QT (QTcF) interval decreased when (R,S)-methadone was replaced by a half-dose of (R)-methadone; the median (interquartile range [IQR]) values were 423 (398-440) milliseconds (ms) and 412 (395-431) ms (P = .06) at days 0 and 14, respectively. Using a univariate mixed-effect linear model, the QTcF value decreased by a mean of -3.9 ms (95% confidence interval [CI], -7.7 to -0.2) per week (P = .04). The QTcF value increased when (R)-methadone was replaced by the initial dose of (R,S)-methadone for 14 days; median (IQR) values were 424 (398-436) ms and 424 (412-443) ms (P = .01) at days 14 and 28, respectively. The univariate model showed that the QTcF value increased by a mean of 4.7 ms (95% CI, 1.3-8.1) per week (P = .006). Substitution of (R,S)-methadone by (R)-methadone reduces the QTc interval value. A safer cardiac profile of (R)-methadone is in agreement with previous in vitro and pharmacogenetic studies. If the present results are confirmed by larger studies, (R)-methadone should be prescribed instead of (R,S)-methadone to reduce the risk of cardiac toxic effects and sudden death.

Impact of an intervention to control risk associated with patient transfer.

QUESTION UNDER STUDY: Hospitals transferring patients retain responsibility until admission to the new health care facility. We define safe transfer conditions, based on appropriate risk assessment, and evaluate the impact of this strategy as implemented at our institution. METHODS: An algorithm defining transfer categories according to destination, equipment monitoring, and medication was developed and tested prospectively over 6 months. Conformity with algorithm criteria was assessed for every transfer and transfer category. After introduction of a transfer coordination centre with transfer nurses, the algorithm was implemented and the same survey was carried out over 1 year. RESULTS: Over the whole study period, the number of transfers increased by 40%, chiefly by ambulance from the emergency department to other hospitals and private clinics. Transfers to rehabilitation centres and nursing homes were reassigned to conventional vehicles. The percentage of patients requiring equipment during transfer, such as an intravenous line, decreased from 34% to 15%, while oxygen or i.v. drug requirement remained stable. The percentage of transfers considered below theoretical safety decreased from 6% to 4%, while 20% of transfers were considered safer than necessary. A substantial number of planned transfers could be "downgraded" by mutual agreement to a lower degree of supervision, and the system was stable on a short-term basis. CONCLUSION: A coordinated transfer system based on an algorithm determining transfer categories, developed on the basis of simple but valid medical and nursing criteria, reduced unnecessary ambulance transfers and treatment during transfer, and increased adequate supervision.

Methods for integration site distribution analyses in animal cell genomes.

The question of where retroviral DNA becomes integrated in chromosomes is important for understanding (i) the mechanisms of viral growth, (ii) devising new anti-retroviral therapy, (iii) understanding how genomes evolve, and (iv) developing safer methods for gene therapy. With the completion of genome sequences for many organisms, it has become possible to study integration targeting by cloning and sequencing large numbers of host-virus DNA junctions, then mapping the host DNA segments back onto the genomic sequence. This allows statistical analysis of the distribution of integration sites relative to the myriad types of genomic features that are also being mapped onto the sequence scaffold. Here we present methods for recovering and analyzing integration site sequences.

E-Cigarettes: A Review of New Trends in Cannabis Use.

The emergence of electronic cigarettes (e-cigs) has given cannabis smokers a new method of inhaling cannabinoids. E-cigs differ from traditional marijuana cigarettes in several respects. First, it is assumed that vaporizing cannabinoids at lower temperatures is safer because it produces smaller amounts of toxic substances than the hot combustion of a marijuana cigarette. Recreational cannabis users can discretely "vape" deodorized cannabis extracts with minimal annoyance to the people around them and less chance of detection. There are nevertheless several drawbacks worth mentioning: although manufacturing commercial (or homemade) cannabinoid-enriched electronic liquids (e-liquids) requires lengthy, complex processing, some are readily on the Internet despite their lack of quality control, expiry date, and conditions of preservation and, above all, any toxicological and clinical assessment. Besides these safety problems, the regulatory situation surrounding e-liquids is often unclear. More simply ground cannabis flowering heads or concentrated, oily THC extracts (such as butane honey oil or BHO) can be vaped in specially designed, pen-sized marijuana vaporizers. Analysis of a commercial e-liquid rich in cannabidiol showed that it contained a smaller dose of active ingredient than advertised; testing our laboratory-made, purified BHO, however, confirmed that it could be vaped in an e-cig to deliver a psychoactive dose of THC. The health consequences specific to vaping these cannabis preparations remain largely unknown and speculative due to the absence of comprehensive, robust scientific studies. The most significant health concerns involve the vaping of cannabinoids by children and teenagers. E-cigs could provide an alternative gateway to cannabis use for young people. Furthermore, vaping cannabinoids could lead to environmental and passive contamination.

��valuation logique de trois (3) campagnes de pr��vention contre le VIH/Sida �� Montr��al

�� l���heure o�� nous ��crivons, l���ONU enregistre 33 million de personnes touch��es par le VIH/Sida. Dans les perspectives actuelles de lutte contre la propagation de la pand��mie, plusieurs moyens peuvent ��tre mis en place : utilisation du condom, d��pistage, ��change de seringues, abstinence��� Mais une question se pose : quels ��l��ments nous influencent dans l���utilisation d���une telle ou telle pratique? Faisant usage des strat��gies de marketing social, les campagnes de pr��vention contre le VIH/Sida mettent l���emphase sur l���empowerment des individus face �� leur prise de risque. Par ce biais, on tente de changer certains comportements et d���en adopter de nouveaux plus s��curitaires pour la sant��. Nous avons ��valu�� ici trois (3) campagnes de pr��vention contre le VIH/Sida �� Montr��al. Le but de cette recherche a ��t�� de distinguer les ��l��ments pouvant faciliter la planification des campagnes dans une perspective de diminution de l���incidence du VIH/Sida. Lors de la prise de d��cisions concernant l'��valuation d'une campagne de pr��vention de lutte contre le VIH/Sida, plusieurs points fondamentaux sont �� consid��rer : la source de l���information, le message, le canal utilis��, les caract��ristiques du r��cepteur et l���effet d��sir�� par la campagne. Ces aspects sont primordiaux dans la prise de conscience de la campagne. Mais attention, ce type d'��valuation n'est pas con��u pour en d��montrer son efficacit��. Notre ��tude nous confirme l���importance de l�����valuation des campagnes de pr��vention aux diff��rents stades de leurs conceptions. Cette recherche nous pousse �� conna��tre les d��tails du programme de pr��vention et ainsi avoir une bonne compr��hension du d��roulement de l'intervention dans une perspective future d���en expliquer l���efficacit��

Protocole de routage bas�� sur des passerelles mobiles pour un acc��s Internet dans les r��seaux v��hiculaires

La rapide progression des technologies sans fil au cours de ces derni��res ann��es a vu na��tre de nouveaux syst��mes de communication dont les r��seaux v��hiculaires. Ces r��seaux visent �� int��grer les nouvelles technologies de l���information et de la communication dans le domaine automobile en vue d���am��liorer la s��curit�� et le confort sur le r��seau routier. Offrir un acc��s Internet aux v��hicules et �� leurs occupants peut sans doute aider �� anticiper certains dangers sur la route tout en rendant plus agr��ables les d��placements �� bord des v��hicules. Le d��ploiement de ce service n��cessite que des messages soient ��chang��s entre les v��hicules. Le routage constitue un ��l��ment crucial dans un r��seau, car d��finissant la fa��on dont les diff��rentes entit��s ��changent des messages. Le routage dans les VANETS constitue un grand d��fi car ces derniers sont caract��ris��s par une forte mobilit�� entra��nant une topologie tr��s dynamique. Des protocoles ont ��t�� propos��s pour ��tendre Internet aux r��seaux v��hiculaires. Toutefois, la plupart d���entre eux n��cessitent un co��t ��lev�� de messages de contr��le pour l�����tablissement et le maintien des communications. Ceci a pour cons��quence la saturation de la bande passante entrainant ainsi une baisse de performance du r��seau. Nous proposons dans ce m��moire, un protocole de routage qui s���appuie sur des passerelles mobiles pour ��tendre Internet aux r��seaux v��hiculaires. Le protocole prend en compte la mobilit�� des v��hicules et la charge du r��seau pour l�����tablissement et le maintien des routes.

The Development, Validation and Implementation of a Broad-Based ADME Genotyping Assay into Research and Clinical Trials

Afin d���adresser la variabilit�� interindividuelle observ��e dans la r��ponse pharmacocin��tique �� de nombreux m��dicaments, nous avons cr���� un panel de g��notypage personnalis��e en utilisant des m��thodes de conception et d�����laboration d���essais uniques. Celles-ci ont pour but premier de capturer les variations g��n��tiques pr��sentent dans les g��nes cl��s impliqu��s dans les processus d'absorption, de distribution, de m��tabolisme et d���excr��tion (ADME) de nombreux agents th��rapeutiques. Bien que ces g��nes et voies de signalement sont impliqu��s dans plusieurs m��canismes pharmacocin��tiques qui sont bien connues, il y a eu jusqu����� pr��sent peu d'efforts envers l�����valuation simultan��e d���un grand nombre de ces g��nes moyennant un seul outil exp��rimental. La recherche pharmacog��nomique peut ��tre r��alis��e en utilisant deux approches: 1) les marqueurs fonctionnels peuvent ��tre utilis��s pour pr��s��lectionner ou stratifier les populations de patients en se basant sur des ��tats m��taboliques connus; 2) les marqueurs Tag peuvent ��tre utilis��s pour d��couvrir de nouvelles corr��lations g��notype-ph��notype. Pr��sentement, il existe un besoin pour un outil de recherche qui englobe un grand nombre de g��nes ADME et variantes et dont le contenu est applicable �� ces deux mod��les d'��tude. Dans le cadre de cette th��se, nous avons d��velopp�� un panel d���essais de g��notypage de 3,000 marqueurs g��n��tiques ADME qui peuvent satisfaire ce besoin. Dans le cadre de ce projet, les g��nes et marqueurs associ��s avec la famille ADME ont ��t�� s��lectionn��s en collaboration avec plusieurs groupes du milieu universitaire et de l'industrie pharmaceutique. Pendant trois phases de d��veloppement de cet essai de g��notypage, le taux de conversion pour 3,000 marqueurs a ��t�� am��lior�� de 83% �� 97,4% gr��ce �� l'incorporation de nouvelles strat��gies ayant pour but de surmonter les zones d'interf��rence g��nomiques comprenant entre autres les r��gions homologues et les polymorphismes sous-jacent les r��gions d���int��r��t. La pr��cision du panel de g��notypage a ��t�� valid��e par l�����valuation de plus de 200 ��chantillons pour lesquelles les g��notypes sont connus pour lesquels nous avons obtenu une concordance > 98%. De plus, une comparaison crois��e entre nos donn��es provenant de cet essai et des donn��es obtenues par diff��rentes plateformes technologiques d��j�� disponibles sur le march�� a r��v��l�� une concordance globale de > 99,5%. L'efficacit�� de notre strat��gie de conception ont ��t�� d��montr��es par l'utilisation r��ussie de cet essai dans le cadre de plusieurs projets de recherche o�� plus de 1,000 ��chantillons ont ��t�� test��s. Nous avons entre autre ��valu�� avec succ��s 150 ��chantillons h��patiques qui ont ��t�� largement caract��ris��s pour plusieurs ph��notypes. Dans ces ��chantillons, nous avons pu valider 13 g��nes ADME avec cis-eQTL pr��c��demment rapport��s et de d��couvrir et de 13 autres g��nes ADME avec cis eQTLs qui n'avaient pas ��t�� observ��s en utilisant des m��thodes standard. Enfin, �� l'appui de ce travail, un outil logiciel a ��t�� d��velopp��, Opitimus Primer, pour aider pour aider au d��veloppement du test. Le logiciel a ��galement ��t�� utilis�� pour aider �� l'enrichissement de cibles g��nomiques pour d'exp��riences s��quen��age. Le contenu ainsi que la conception, l���optimisation et la validation de notre panel le distingue largement de l���ensemble des essais commerciaux couramment disponibles sur le march�� qui comprennent soit des marqueurs fonctionnels pour seulement un petit nombre de g��nes, ou alors n���offre pas une couverture ad��quate pour les g��nes connus d���ADME. Nous pouvons ainsi conclure que l���essai que nous avons d��velopp�� est et continuera certainement d�����tre un outil d���une grande utilit�� pour les futures ��tudes et essais cliniques dans le domaine de la pharmacocin��tique, qui b��n��ficieraient de l'��valuation d'une longue liste compl��te de g��nes d���ADME.