Le sepsis, cause majeure de décès en réanimation, entraîne des altérations immunitaires associées à un risque augmenté de décès et d'infections secondaires. En particulier, les lymphocytes T (LT) de patients présentent des altérations phénotypiques et fonctionnelles caractéristiques d'un état d'épuisement. Pour améliorer leur réponse, l'IL-7 est actuellement proposée comme immunostimulant. Son récepteur existe sous différentes formes: protéines membranaire (CD127) et soluble (sCD127) et différents transcrits. Leur expression n'a été que peu étudiée dans le sepsis. De plus, dans d'autres contextes cliniques, le phénotype CD127lowPD1high est proposé comme marqueur d'épuisement des LT mais n'a jamais été évalué dans le sepsis. L'objectif de ce projet était d'évaluer l'expression du récepteur de l'IL-7 comme biomarqueur dans le sepsis, d'étudier sa régulation chez les patients et la présence du phénotype CD127lowPD-1high en lien avec les altérations des LT. L'objectif était également de mettre au point un modèle ex vivo reproduisant les altérations intrinsèques aux LT de patients. Nous avons confirmé l'intérêt de la concentration plasmatique de sCD127 comme marqueur de mortalité chez les patients de réanimation. Nous avons mis en évidence l'association de l'expression des transcrits IL7R correspondant à des formes solubles avec la mortalité, ainsi que leur régulation intrinsèque dans les LT. Enfin, la proportion augmentée de LT CD127lowPD-1high chez les patients en choc septique en fait également un candidat potentiel comme marqueur spécifique des altérations des LT. Dans un deuxième temps, nous avons développé un modèle d'altérations induites par l'activation à partir de LT purifiés de volontaires sains reproduisant les altérations des LT de patients en choc septique. Ce modèle suggère un rôle de l'activation des LT dans le développement de leurs altérations et pourrait permettre d'étudier de nouveaux aspects de la physiopathologie des LT dans le sepsis / Sepsis is the leading cause of death in intensive care units (ICU). Septic patients develop immune dysfunctions that are associated with an increased risk of death and secondary infections. In particular, patients’ T lymphocytes present phenotypic and functional alterations characteristic of exhaustion. To restore these alterations, IL-7 is currently proposed as an immunostimulant. Its receptor exists in different forms: a membrane (CD127) and a soluble (sCD127) proteins and different transcripts. The IL-7 receptor expression has not been studied in sepsis. Moreover, the CD127lowPD-1high phenotype is proposed as a T cell exhaustion marker in other clinical contexts but has never been explored in sepsis.In this context, the aim of this project was first to evaluate the expression of the IL-7 receptor as a biomarker in sepsis, to study its regulation in patients and the CD127lowPD-1high phenotype in parallel with T cell alterations in septic shock patients. Secondly, the objective was to develop an ex vivo model reproducing phenotypic and functional T cell alterations observed in patients.We confirmed the role of sCD127 plasmatic concentration as a marker of mortality in septic shock and more generally in ICU patients. In addition, we demonstrated the association of the expression of the IL7R transcripts, corresponding to soluble forms with mortality as well as the intrinsic regulation of their expressions in T lymphocytes. Finally, we found an increased proportion of CD127lowPD-1high T cells in septic shock patients. This population could also be evaluate as a potential specific marker of T lymphocyte alterations. Secondly, we developed an ex vivo model of alterations induced by activation of purified T cell from healthy volunteers reproducing alterations observed in patients’ T cells. This model suggests a role of T cell activation in the development of their alterations and could be used to explore new aspects of T cell alterations pathophysiology in sepsis

A diálise peritoneal constitui uma importante opção terapêutica para o paciente com doença renal crônica (DRC) em estágio 5. Entretanto, a médio-longo prazo, alterações morfofuncionais relacionadas a vários fatores, como bioincompatibilidade das soluções de diálise e infecções peritoneais, entre outros, estabelecem um processo inflamatório e fibrótico na membrana peritoneal, levando à perda da eficiência deste método dialítico. O estado urêmico destes pacientes é um agravante, pois intensifica o processo inflamatório da membrana peritoneal. Estratégias terapêuticas que desacelerem o processo de fibrose da membrana peritoneal dos pacientes com DRC em diálise são de extrema importância. Neste contexto, o presente estudo teve como objetivo estabelecer um modelo de peritonite fibrosante associado à DRC com uremia, que mimetiza a situação clínica, e analisar, neste modelo, o efeito de duas moléculas antifibróticas, o tamoxifeno (TAM) e a BMP-7 (bonemorphogenic protein-7). A DRC com uremia foi induzida em ratos Wistar através da administração de dieta rica em adenina, por um período de 30 dias. Nos últimos 15 dias, com o estado de uremia já estabelecido, os animais receberam injeções intraperitoneais de gluconato de clorexidina para a indução da fibrose peritoneal (FP). Os tratamentos com tamoxifeno (10mg/Kg/dia, por gavagem) e BMP-7 (30ug/Kg, injeções intraperitoneais a cada 3 dias) foram iniciados junto com a indução da fibrose peritoneal. Foram formados 6 grupos experimentais: CONTROLE, animais normais; DRC, animais com doença renal crônica; FP, animais com fibrose peritoneal; DRC/FP, animais com DRC e FP mimetizando a situação clínica; DRC/FP+TAM, animais com DRC e FP tratados com tamoxifeno; e DRC/FP+BMP7, animais com DRC e FP tratados com BMP-7. Durante os 30 dias de seguimento do estudo foram verificados o peso, a pressão arterial, ureia e creatinina séricas dos animais. Ao término deste período os animais foram sacrificados e o peritônio foi removido e submetido às análises: a) histológica, para avaliar o grau de espessamento (Tricrômio de Masson); b) imunohistoquímica, para localizar e quantificar a presença de células inflamatórias (macrófagos, linfócitos T), miofibroblastos (?-actina) e a atividade de proliferação celular (PCNA); c) análise de citocinas pró-inflamatórias (TNF-alfa, IL-1beta e IL-6) no tecido peritoneal tanto em nível de RNAm, através de PCR em tempo real, como também em nível proteico, através de multiplex; d) PCR em tempo real para determinar a expressão dos componentes da matriz extracelular (colágeno III e fibronectina) e de fatores fibrogênicos (TGF-beta1 e FSP-1). Além disso, com o objetivo de estudar a possível via de sinalização associada à fibrose peritoneal, foi analisada a expressão de SMAD 3 e SMAD 7 no peritônio através de PCR em tempo real e imunohistoquímica para SMAD 3 fosforilada. Por fim, a função peritoneal foi analisada através do teste de ultrafiltração e massa transferida de glicose (MTG). Os dados da evolução ponderal mostraram que, enquanto os animais dos grupos Controle e FP tiveram um ganho de peso significativo em relação ao primeiro dia do protocolo (28% e 18%, respectivamente), os animais dos demais grupos perderam peso significativamente, em média, 26% em relação ao primeiro dia. Todos os animais que receberam dieta rica em adenina e que, portanto, desenvolveram DRC, apresentaram hipertensão arterial, detectada nos dias 15 e 30 do estudo (média de 173mmHg no dia 15 e 172mmHg no dia 30). Confirmando o estabelecimento de DRC com uremia, os animais que receberam dieta rica em adenina apresentaram níveis séricos significativamente elevados de uréia (em média 170 mg/dL no dia 15 e 286 mg/dL no dia 30) e creatinina (média de 0,97 mg/dL no dia 15 e 1,82 mg/dL no dia 30). Os tratamentos com TAM e BMP-7 não influenciaram significativamente nestes parâmetros. A análise da membrana peritoneal dos animais dos grupos experimentais FP e DRC/FP revelou um espessamento significativo da membrana peritoneal (130 ± 33um e 132 ± 26?m, respectivamente vs 36 ± 2um e 27 ±6 um nos grupos CONTROLE e DRC; p < 0,001) bem como a presença de células inflamatórias. Os tratamentos com TAM e BMP7 foram eficazes em proteger a membrana peritoneal contra o espessamento (42 ± 2?m e 53 ± 7?m, respectivamente; p < 0,001 vs DRC/FP) e contra o infiltrado inflamatório. Além disso, no que se refere aos miofibroblastos, células efetoras da fibrogênese detectadas pela marcação de ?-actina, foi encontrado uma marcação significativa de alfa-actina no peritônio dos grupos FP e DRC/FP (p < 0,01 vs CONTROLE), sendo que os tratamentos com TAM e BMP7 protegeram o peritônio da proliferação maciça de miofibroblastos, confirmada pela expressão de PCNA de forma significativa. Com relação à detecção de citocinas pró-inflamatórias, a análise por PCR em tempo real nos animais do grupo DRC mostrou um aumento significativo da expressão no peritônio de TNF-alfa e IL-1beta comparado ao grupo CONTROLE. A expressão dessas citocinas também se mostrou aumentada no peritônio dos animais dos grupos e DRC/FP. Os tratamentos com TAM e BMP7 reduziram a expressão de TNF-alfa e IL-1beta de forma significativa em relação ao grupo DRC/PF (p < 0,01). O reflexo destes resultados pode ser observado com o ensaio por multiplex em que a presença dessas citocinas em sua forma proteica foi encontrada em quantidades significativas no peritônio dos animais com FP e uremia associada à FP em relação ao grupo CONTROLE. Os tratamentos com TAM e BMP7 reduziram a presença dessas citocinas de forma significativa (p < 0,01 vs DRC/FP). Como esperado, a presença de RNAm de componentes da matriz extracelular foi significativamente maior nos grupos FP e DRC/FP, comparados ao grupo CONTROLE. De fato, a expressão de colágeno III foi maior nos grupos FP e DRC/FP (3,4 ± 1 e 10,3 ± 2,4 UI, respectivamente; p < 0,01 vs CONTROLE), bem como de fibronectina nos grupos (6,5 ± 0,8 e 29,2 ± 1 UI, respectivamente; p < 0,01 vs CONTROLE). Os animais tratados com TAM e BMP7 apresentaram uma diminuição significativa tanto da expressão de colágeno III (1,5±0,9 e 0,2±0,1 UI, respectivamente; p < 0,01 vs DRC/FP), como da expressão de fibronectina (6,6±1,2 e 9,7±0,5 UI, respectivamente; p < 0,01 vs DRC/FP), confirmando um efeito anti-fibrótico dessas drogas. Ainda, enquanto nos grupos FP e DRC/FP a expressão de TGF-? foi significativamente maior em relação ao grupo CONTROLE (13,2±0,9 e 30,3±0,1 UI, respectivamente; p < 0,01), TAM e BMP7 diminuíram a expressão de TGF-beta (17,7±0,2 e 16,2±0,1 UI, respectivamente; p < 0,01 vs DRC/FP). Padrão similar foi encontrado com a expressão do RNAm para FSP-1 em que houve um aumento significativo da expressão desse gene nos grupos FP e DRC/FP, com significativo bloqueio nos animais tratados com TAM e BMP7 (p < 0,01 vs DRC/FP). Com relação à análise das vias de sinalização possivelmente envolvidas no processo, a expressão de SMAD 3 foi significativamente maior nos grupos FP e DRC/FP (3,7±0,3 e 4,6±0,3 UI, respectivamente) em relação aos grupos CONTROLE e DRC (1±0,2 e 1,3±0,6 UI, respectivamente; p < 0,01). Os tratamentos com TAM e BMP7 diminuíram a expressão da SMAD 3 no peritônio (1,1±0,6 e 1,1±0,7 UI, respectivamente; p < 0,01 vs DRC/FP). Contudo TAM e BMP7 aumentaram significativamente a expressão de SMAD 7 (2,8±0,5 e 3,7±0,5 UI, respectivamente; p < 0,01 vs DRC/FP), uma proteína contra reguladora da via do TGF-beta, capaz de bloquear a expressão de fatores pró-fibróticos bem como de fatores inflamatórios. Finalmente, os grupos tratados TAM e BMP7 tiveram a função do peritônio preservada quando comparados aos grupos FP e DRC/FP, verificado pela manutenção da capacidade de ultrafiltração e de reduzir a MTG. Em resumo, tamoxifeno e BMP7 foram capazes de bloquear o espessamento da membrana peritoneal, proteger o peritônio contra a infiltração de células inflamatórias e de miofibroblastos, além de diminuir a proliferação celular no peritônio. Estes tratamentos também foram eficazes em diminuir significativamente a expressão dos fatores pró-fibróticos e das citocinas inflamatórias. Com relação às SMADs, os tratamentos com TAM e BMP7 foram eficazes em bloquear a expressão de SMAD 3, bem como aumentar a expressão de SMAD 7. Os resultados do presente estudo sugerem que tamoxifeno e BMP7 protegem o peritônio no modelo de fibrose peritoneal desenvolvido em ratos com DRC e uremia, possivelmente devido aos seus efeitos anti-inflamatórios e anti-fibróticos / Peritoneal dialysis is an important therapeutic option for patients with stage 5 chronic kidney disease (CKD). However, at medium and long term, morphological and functional changes related to various factors such as bioincompatibility of dialysis solutions and peritoneal infections, among others, establish an inflammatory and fibrotic process in the peritoneal membrane, leading to a loss of dialysis efficiency. The uremic state of these patients aggravates this situation because it intensifies inflammation of the peritoneal membrane. Therapeutic strategies that slow the process of fibrosis of the peritoneal membrane of patients with CKD on dialysis are extremely important. In this context, the present study aimed to establish a model of peritoneal fibrosis associated with CKD with uremia, that mimics the clinical situation, and analyze the effect of two antifibrotic molecules, tamoxifen (TAM) and the BMP7 (bone morphogenic protein-7), in the proposed model. CKD with uremia was induced in male Wistar rats by adenine in the diet during a period of 30 days. After 15 days, with the state of uremia already established, animals received intraperitoneal injections of chlorhexidine gluconate, for the induction of peritoneal fibrosis (PF). Treatment with TAM (10mg/Kg/day by gavage) and BMP7 (30ug/Kg, intraperitoneal injections every 3 days) were initiated along with the induction of peritoneal fibrosis. Six groups were induced: CONTROL, normal animals; CKD, animals with chronic kidney disease; PF, animals with peritoneal fibrosis; CKD / PF, animals with CKD and PF mimicking the clinical situation; CKD / PF + TAM, animals with CKD and PF treated with tamoxifen; and CKD / PF + BMP7, animals with CKD and PF treated with BMP7. During 30 days of the follow-up study, weight, blood pressure, serum urea and creatinine of the animals were verified. At the end of this period, the animals were sacrificed and the peritoneum was removed and subjected to the following analysis: a) histology, to assess the degree of thickening (Masson\'s Trichrome); b) immunohistochemistry, to locate and quantify the presence of inflammatory cells (macrophages, T lymphocytes), myofibroblasts (alfa-smoth muscle actin) and cell proliferation activity (PCNA); c) analysis of pro-inflammatory cytokines (TNF-alfa, IL-1beta and IL-6) both in peritoneal tissue mRNA level through real time PCR as well as on protein level by multiplex; d) real-time PCR to determine the expression of extracellular matrix components (collagen III and fibronectin) and fibrogenic factors (TGF-beta and FSP-1). Furthermore, in order to study the possible signaling pathway associated to peritoneal fibrosis, the expression of SMAD 3 and SMAD 7 in the peritoneum was analyzed via real-time PCR and immunohistochemistry for phosphorylated SMAD 3. Finally, the peritoneal function was assessed by ultrafiltration and mass transferred glucose test (MTG). Data from weight gain showed that while animals from CONTROL and PF groups had significant weight gain (28% and 18% respectively) compared to the first day of protocol, the animals of other groups lost weight significantly, on average, 26% compared to the first day. All animals that received diet rich in adenine developed CKD presented by hypertension, detected on days 15 and 30 of the study (average of 173mmHg and 172mmHg respectively). Confirming the establishment of CKD with uremia, the animals that received diet rich in adenine showed serum urea (average 170 mg/dL on day 15 and 286 mg/dL on day 30) and creatinine (average of 0.97 mg/dL on day 15 and 1.82 mg/dL on day 30) significantly higher in the 15th and 30th days. Treatments with TAM and BMP7 did not influence these parameters. The peritoneal membrane analysis of PF and CKD/PF experimental groups showed a significant thickening of the peritoneal membrane (130 ± 33?m and 132 ± 26?m, respectively; p < 0.001 vs 36 ± 2um and 27 ± 6?m in CONTROL and CKD; p < 0.001) with presence of inflammatory cells. The treatments with TAM and BMP7 were effective in protecting the membrane against the thickening (42 ± 2um and 53±7um, respectively; p < 0.001 vs CKD/PF) and inflammatory infiltrate. Furthermore, with regard to myofibroblasts, effectors cells in the fibrogenesis process detected by alfa-SMA presence, it was found a signicantly expression in the peritoneum of PF and CKD/PF (p < 0.01 vs CONTROLE), and the treatment with TAM and BMP7 significantly protected against the presence of myofibroblasts confirmed by the significantly expression of PCNA. With regard to the detection of pro-inflammatory cytokines, analysis by real-time PCR in the animals of CKD group showed a significant increase in TNF-alfa expression in the peritoneum and IL-1beta compared to the CONTROL group. The expression of these cytokines was also increased in the peritoneum of the CKD/PF group. The treatment with TAM and BMP7 significantly reduced TNF-alfa and IL-1beta expression compared to CKD/PF group (p < 0.01).The repercussion of these results can be observed with the multiplex test in which the presence of these cytokines in its protein form were found in significant quantities in the peritoneum of the animals with uremia associated with PF compared to CONTROL group. The treatment with TAM and BMP7 significantly reduced the presence of these cytokines (p < 0.01 vs CKD/PF). As expected, the mRNA expression of extracellular matrix components was significantly elevated in the PF group and CKD/PF compared to the control group. Indeed, collagen III mRNA expression was higher in PF and CKD/PF group (3.4 ± 1 and 10.3 ± 2.4 UI, respectively; p < 0.01 vs CONTROL) as well as fibronectin (6.5 ± 0.8 and 29.2 ± 1 UI; respectively; p < 0.01 vs CONTROL). The animals treated with TAM and BMP7 significantly blocked the expression of collagen III (1.5 ± 0.9 and 0.2 ± 0.1 UI, respectively; p < 0.01 vs CKD/PF) and fibronectin (6.6±1.2 and 9.7±0.5 UI, respectively; p < 0.01 vs CKD/PF). Further, while in the PF and CKD/PF groups the expression of TGF-beta (13.2 ± 0.9 UI and 30.3 ± 0.1 UI, respectively; p < 0.01) was significantly higher compared to the CONTROL group, TAM and BMP7 decreased the expression of TGF-beta (17.7 ± 0.2 UI and 16.2 ± 0.1UI, respectively; p < 0.01 vs CKD/PF). A similar trend was found with the expression of FSP-1 mRNA with an increase in expression of this gene in PF and CKD/PF groups (p < 0.01 vs CONTROL), with significant blockage in the animals treated with TAM and BMP7 (p < 0.01 vs CKD/PF). Regarding the analysis of signaling pathways possibly involved in the process, SMAD 3 expression was significantly higher in PF and CKD/PF groups (3.7 ± 0.3 UI and 4.6 ± 0.3 UI, respectively) compared to groups CONTROL and CKD (1 ± 0.2 UI and 1.3 ± 0.6 UI, respectively; p < 0,01). Treatments with TAM and BMP7 decreased the expression of SMAD 3 in the peritoneum (1.1 ± 0.6 UI and 1.1 ± 0.7 UI, respectively; p < 0.01 vs CKD/PF). Yet TAM and BMP7 significantly increased the expression of SMAD 7 (2.8 ± 0.5 and 3.7 ± 0.5, respectively; p < 0.01 vs CKD/PF), a regulatory TGF-beta protein capable of blocking the expression of pro-fibrotic and inflammatory factors. Finally, the treated groups had the function of the peritoneum preserved when compared to the PF and CKD / PF groups, checked through the maintenance of the ultrafiltration capacity and reducing MTG. In summary, the animals treated with TAM and BMP7 had the peritoneum protected from thickening, inflammatory infiltrate, presence of myofibroblasts and cellular proliferation. The treatments were also effective in significantly reduce the expression of pro-fibrotic factors and inflammatory cytokines. Regarding SMADs, treatments with TAM and BMP7 were effective in blocking the expression of SMAD 3 and increase SMAD 7 expression. The results of this study suggest that tamoxifen and BMP7 protected the peritoneum in an experimental model of peritoneal fibrosis developed in uremic rats with CKD, possibly due to their anti-inflammatory and anti-fibrotic properties

La decisión de ocuparme de un tema que abarca al mismo tiempo dos personajes distintos, es imputable principalmente a mi fuerte interés por el patronazgo femenino renacentista, y en segundo lugar, al hecho que las dos figuras tratadas, aunque muy estudiadas singularmente, nunca han estado realmente profundizadas en cuanto cuñadas y, por ese motivo, fuertemente afines. Analizando la bibliografía relativa a la relación existida entre Isabella d’Este y Lucrezia Borgia, de hecho resultada más bien lagunosa, y encontrando en archivo una surtida correspondencia que les pertenece, he pensado en intentar alumbrar aquella parte de su historia con una pieza más, demorándome sobre la manera con el que se relacionaron a la hora de afrontar aquellos aspectos de su vida que las obligabas a confrontarse. Teniendo en cuenta que la bibliografía existente concuerda en aseverar que Isabella tenía en gran antipatía su cuñada, mi curiosidad, ya desde el principio, consistía en averiguar si realmente Isabella era tan envidiosa de Lucrezia, y si así fuera, cómo se relacionó con ella a lo largo de los diez y siete años en el que fueron parientes y con cuales consecuencias bajo varios puntos de vista. El resultado de la investigación me ha llevado a individuar tres grandes temas que las vieron, con factores distintos, estrechamente conectadas; el primer tema es aquello ligado al patronazgo de cultura que, puesta la posición de Isabella en querer ser recordada cual única primera dama de su tiempo, las ve netamente en contraposición, con, por un lado, intelectuales y humanistas empeñados en tejer las laudes de ambas, y por otro, la evidente aversión demostrada por Isabella hacia Lucrezia, que en vano intenta establecer un dialogo erudito con la cuñada. En segundo lugar, la investigación ha evidenciado inesperados puntos de unión por lo que se refiere al tema de la religión y sus distintas facetas, de que estaba cargada toda la sociedad renacentista; partiendo por dos posiciones muy distintas, es posible ver los diferentes aproches de ambas en algunas circunstancias claves ligadas al mundo eclesiástico. En fin, el enfoque se ha desplazado sobre los asuntos estrechamente inherentes a su posición en seno a sus familias, resaltando temas familiares y de gestión del propio deber cuales señoras de sus estados. La investigación ha conducido a resultados interesantes e inesperados, sobre todo cuando la historia oficial se ha mezclado con la historia intima de las dos mujeres, ligada a su intercambio epistolar. Al término de esta investigación que ha abarcado un largo lapso de historia a caballo entre siglo XV y siglo XVI Y Analizando el intercambio epistolar, notamos que la correspondencia está sostenida principalmente por Lucrezia, que parece sufrir de una forma de grafomanía unida a su necesidad de resultar agradecida a Isabella. Como visto, por su parte Isabella resulta haber escrito pocas cartas a Lucrezia, dejando frecuentemente que fuese un miso a transmitirle el mensaje, y los temas que toca son a menudo muy convencionales, denotando cierta frialdad hacía ella. Una particularidad que sobresale es que si entre Isabella y Alfonso la correspondencia resulta constante y rica de temas de grande interés cultural y artístico, o sea los temas favoritos de Isabella, con Lucrezia, este tipo de contenido falta completamente. La única ocasión en la que tratan de un artista en su correspondencia es justo al principio, cuando Lucrezia todavía se ilude, considerando la extracción humanística e intelectual de ambas, de poder empezar una relación de cualidad con su cuñada, que de hecho hará exactamente lo contrario de lo que ella espera, escribiendo poco y tocando temas de escaso interés. En conclusión, podemos decir que en un periodo de grande fervor cultural y verdadero receptáculo de ilustres personajes, dos damas de dos grandes familias se han distinguido, y que con más o menos fortuna critica han llegado hasta nuestro días, casi como dos heroínas. Detrás de la imagen que nos han dejado, sin embargo se celan, antes que todo, dos mujeres extraordinarias, que han sabido distinguirse en una sociedad mandada por los hombres, imponiéndose cuando la situación lo requería, y utilizando también astucias típicamente femeninas para obtener los resultados esperados. A través de su intercambio epistolar sobresalen totalmente estas dos mujeres; empeñadas a veces con la rutina cotidiana, a veces con problemas a menudo más grandes que ellas, y aunque podemos tal vez lamentar de la escasa complicidad demostrada, podemos por otra parte verlas en toda su humanidad y fragilidad, reconociendo en ellas defectos y valor, y seguramente apreciándolas ambas, por lo que han sido y también por lo que no han sido / ENGLISH ABSTRACT The decision to deal with a subject that covers simultaneously two different characters, is attributable mainly to my strong interest in Renaissance female patronage, and secondly, at the fact that the two figures treated, although very studied singularly, have never been really deepened as sisters in law and, for that reason, strongly related. Analyzing the literature on the relationship existed between Isabella d'Este and Lucrezia Borgia, in fact turned out rather lacunose, and finding a stocked correspondence on file that belongs to them, I thought of trying to illuminate that part of their history with another piece, linger myself on the manner in which they were related to tackling those aspects of their life that obligate them to confront. Given that the literature is consistent in asserting that Isabella had a great dislike to her sister in law, my curiosity, from the beginning, was to find out if you really Isabella was so jealous of Lucrezia, and if so, how he related to her over the seventeen years in which they were relatives and which been the consequences under various viewpoints. The result of research has led me to single out three issues that saw, with different, closely connected factors; the first issue is that linked to patronage of culture, considering Isabella in the position of wanting to be remembered that only first lady of her time, seen clearly in contrast, with one hand, intellectuals and humanists determined to weave the praises of both, and secondly, the apparent aversion demonstrated by Isabella to Lucrezia, who vainly tries to establish a dialogue with the sister in law. Second, research has shown unexpected attachment points so it addresses the issue of religion and its various facets, that was charged full Renaissance society; starting with two very different positions, it is possible to see the different approaches of both in some key factors linked to the ecclesiastical world. Finally, the focus has shifted on closely associated with their position in matters within their families, highlighting family management and ladies of their states which own duty issues. The research has led to interesting and unexpected results, especially when the official story has been mixed with the intimate story of two women, linked to their epistolary exchange. Upon completion of this investigation that has spanned a long period of history straddling and analyzing the exchange of letters XV and XVI century, we note that the correspondence is mainly supported by Lucrezia, who seems to suffer from a form of graphomania due to her need to be grateful to Isabella. As seen, meanwhile Isabella is having written few letters to Lucrezia, often leaving it to be a messenger to convey the message, and the issues it raises are often very conventional, denoting a certain coldness from her. One feature that stands out is that if between Isabella and Alfonso correspondence is consistent and rich themes of great cultural and artistic interest, that is the favorite themes of Isabella, with Lucrezia, this type of content is completely missing. The only time they treat an artist in their correspondence is just at the beginning, when Lucrezia deluding herself, still considering the humanistic and intellectual extraction of both, she can begin a relationship of quality with her sister in law, who in fact do exactly the contrary to what she expected, writing and playing some topics of little interest. In conclusion, we can say that in a period of great cultural fervor and true cradle of famous people, two women of illustrious families have distinguished, and with more or less success criticism have come down to our days, almost as two heroines. Behind the image we have left, however conceal, first of all, two extraordinary women who have managed to distinguish itself in a society controlled by men, beating when the situation required, and also using typically feminine wiles to get the expected results. Through their epistolary exchange these two women stand completely; engaged sometimes daily routine, sometimes with problems bigger than they often, and although we may lament the lack proven complicity, can moreover see them in all their humanity and frailty, recognizing these shortcomings and value, and surely appreciate them both for what they have been and what they have not been

RESUMEN El interés fundamental de esta tesis se ha basado en el análisis del texto como estrategia creativa en las distintas prácticas artísticas y sociales dentro del espacio público, siendo el lenguaje objeto primordial del estudio de dicho fenómeno en si. El lenguaje es un instrumento del pensamiento, y que se revela como uno de los medios más efectivos para expresar ideas por medio del proceso comunicativo y como vehículo para la información hacia el espectador. A la vez, es el sistema comunicativo más convencional. Así, como creemos apreciar, la tendencia en las artes plásticas y visuales a usar el texto como recurso legitimador desde el siglo XX hasta el momento actual, como estrategia lícita, conceptual, que implica todo un trabajo de estructuración y orden a nivel de proyección de la obra artística, y que atiende a factores sobre la complejidad y la operatividad de la misma, diferenciándose totalmente de los conceptos más inmediatos de acción como es el de la táctica, erróneamente utilizado como sinónimo de la estrategia. Revelándose aquí, el concepto de estrategia como recurso conceptual primordial en aquellas producciones que emplean el uso del texto como estrategia para la creación. En el estudio del lenguaje, nos centraremos en la retórica visual un modo de análisis que nos puede permitir, en distintos soportes, o en distintos medios- artes plásticas y visuales, cine, publicidad- articular significante y significado, o dicho de otro modo, forma y contenido, con el objetivo de comprender mejor una obra artística desde el punto de vista del espectador o como creadores de la misma. A la vez que, en cuanto al uso de un método o de una estrategia, se puede apreciar que el uso del pensamiento por medio del lenguaje y sus distintas figuras retóricas, pueden servir como estrategias de creación. Palabras claves: Texto, estrategia creativa, retórica visual, espacio público, prácticas sociales del arte OBJETIVOS DE LA TESIS: Dentro de los objetivos de esta tesis que estudia el uso del texto como estrategia creativa nos hemos propuesto: - Estudiar por medio de un recorrido histórico el uso del texto como estrategia creativa a lo largo de la historia de las artes plásticas y visuales. - Observar los distintos aspectos que se dan en la fusión entre la imagen y el texto, así como la relación que se establece entre la propia tipografía con la semiótica visual como la importancia de las figuras retóricas como figuras comunicativas en los distintos discursos artísticos. - Analizar aquellas características que unen a las distintas prácticas artísticas que usan el texto como estrategia creativa. - Comprender la importancia del término de estrategia, como procedimiento conceptual que implica todo un trabajo de estructuración y orden a nivel de proyección de la obra artística, y que atiende a factores sobre la complejidad y la operatividad de la misma - Establecer por medio del análisis de estas distintas prácticas los distintos soportes o vehículos donde insertan el texto. - Observar la importancia del texto como estrategia creativa en las distintas prácticas sociales dentro del análisis de los autores seleccionados, así como el uso del lenguaje influirá determinantemente en el significado general de las obras de dichos autores. METODOLOGÍA: La metodología utilizada se ha fundamentado, en una de sus partes, en el libro Investigación educativa. Fundamentos y metodología de Arnal, Del Rincón y Latorre (1992), así como en Técnicas de Investigación en Ciencias Sociales (Latorre & Sanz, 1995). Se enmarcará dentro de la metodología de investigación social de tipo cualitativo/constructivista. CONCLUSIONES: El lenguaje es un instrumento del pensamiento, y que se revela como uno de los medios más efectivos para expresar ideas por medio del proceso comunicativo y como vehículo para la información hacia el espectador. El uso del texto como estrategia creativa será uno de los recursos formales y conceptuales que los artistas incorporarán en sus obras caracterizando a una gran parte de la historia de la producción artística del siglo XX, dando paso a que se reconozca el uso del lenguaje como una herramienta más de expresión, y que se ontemple el hecho artístico como el arte de las ideas o del pensamiento. / ABSTRACT The main focus of this thesis is based on the analysis of the text as a creative strategy in the different artistic and social practices within the public space, being the primary language object of study of this phenomenon itself. Language is an instrument of thought, and who reveals himself as one of the most effective means to express ideas through the communication process and as a vehicle for information to the viewer. At the same time, it is the conventional communication system. So, as we see, the trend in plastic and visual to use the text as a legitimizing resource from the twentieth century to the present, as lawful, conceptual arts strategy, which involves a whole structure and work order projection level the artistic work, and serving factors on the complexity and functionality of the same, differing entirely on immediate action concepts such as tactics, wrongly used as a synonym for the strategy. Revealing here, the concept of strategy mainly conceptual resource in those productions that employ the use of text as a strategy for creation. In the study of language, we will focus on the visual rhetoric a mode of analysis that can allow us, in different media or in different media- visual arts, film, advertising-articular signifier and signified, or in other words, form and content in order to better understand a work of art from the point of view of the spectator or as creators of it. While, in the use of a method or strategy, it can be seen that the use of thought through language and its various tropes, can serve as building strategies. Keywords: Text, creative strategy, visual rhetoric, public space, social practices of art OBJECTIVES OF THE THESIS: Among the objectives of this thesis studies the use of text as a creative strategy we have proposed: - Study through a historical text use as creative strategy throughout the history of the visual arts. - Observe the different aspects that occur in the fusion of image and text, as well as the relationship established between the font itself with the visual semiotics as the importance of rhetorical figures as communicative figures in different artistic discourses. - Analyze the characteristics that unite different artistic practices that use the text as a creative strategy. - Understand the importance of the term strategy as a conceptual process that involves all work order level structure and projection of artistic work, and serving factors on the complexity and functionality of the same - Establish through analysis of these different practices or different media vehicles where the inserted text. - Note the importance of the text as a creative strategy in different social practices in the analysis of the selected authors and language use a determining influence on the general meaning of the works of these authors. METHODOLOGY: The methodology is based on one of its parts, in the book Educational research. Fundamentals and methodology of Arnal, Del Rincon & Latorre (1992) and Technical Research in Social Sciences (Latorre & Sanz, 1995). It will be framed within the social research methodology qualitative / constructivist. CONCLUSIONS: Language is an instrument of thought, and who reveals himself as one of the most effective means to express ideas through the communication process and as a vehicle for information to the viewer. The use of text as a creative strategy will be one of the formal and conceptual resources that artists incorporate in their work characterizing much of the history of twentieth century art production, leading to the use of language is recognized as a tool of expression, and that the artistic act as the art of ideas or thought is contemplated.

El objetivo de esta investigación es fijar el campo semántico del concepto de representación en el momento de su origen (siglos XVII y XVIII). Pretendemos identificar las diversas esferas de la experiencia sobre las que se ha fundamentado esta construcción conceptual y, al mismo tiempo, describir las mutaciones que ha sufrido, hasta su crisis. Estas esferas son: artística, epistemológica y jurídico-política. Nuestra hipótesis supone que la nueva concepción del poder político contractualista depende de la metáfora de la representación teatral. Trataremos de mostrar cómo la vieja metáfora del Theatrum Mundi, liberada del sistema teológico, se va a transformar en el Barroco en una forma de sociedad espectacular. El ciudadano se convierte en espectador tanto en el teatro como en la sociedad y en la política. Nuevas relaciones de poder y nuevos sujetos (la burguesía y las masas urbanas), pero también nuevos espacios ciudadanos: reales y abiertos (plazas y jardines), y espacios virtuales y cerrados (teatros). Ese complejo estético-político tiene en la ópera cómica, desarrollada durante el clasicismo, el signo distintivo de la Ilustración y de la burguesía. Los textos han sido tratados por una metodología hermenéutica haciendo una nueva lectura de todos aquellos documentos originales que presentaban una perspectiva de esa metáfora escénica. La problemática actual de la representación política se ha tratado desde el método genealógico usando tres perspectivas que constituyen sendas figuras conceptuales. La primera es la que presenta las tres disciplinas por las que conocemos la representación en Occidente: la teoría política, la epistemología y la estética. La segunda hace referencia a los personajes representativos: el político, el científico y el artista. Finalmente, la representación cultivada por estos tres personajes constituye el espectáculo que se ofrece al público. El espectador “actúa” en la representación como el representado. El espectador (público-masa), constituye la tercera figura conceptual o perspectiva. El resultado de este tratamiento nos permite ver cómo, gracias a esa forma de diferenciación social basada en la distinción o gusto estético, frente a la ópera y otras manifestaciones artísticas, se configuraba un sistema espectacular absolutamente vertical. Las altas culturas se presentaban a aquellos individuos distinguidos, convertidos en personajes representativos (alta burguesía). La moderna sociedad de clases reinventa la desigualdad a través de la cultura. En las reflexiones sobre el arte dramático hemos descubierto las claves para la comprensión del moderno hombre público en tanto que personaje representativo. El espacio público se ha transformado en un escenario (El gran teatro del mundo) y se presenta bajo la forma de la representación. La sociedad y la ciudad se han convertido en una escenografía para el espectáculo; el ciudadano es el público y para él se procede a organizar la compleja representación. Se diseña la figura profesional del representante de los asuntos públicos: el político representativo, casi de la misma manera que hay un protagonista de la representación en el teatro: el actor. Dentro de las conclusiones constatamos que la representación no ha entrado en crisis, como viene anunciado en arte y política, más bien al contrario, se ha reforzado, y la moderna red global está organizada por los sistemas mediáticos que gestionan la opinión pública y estimulan el consumo espectacular de las masas. Hemos identificado, en diálogo con Foucault, que la crisis no ha afectado al sistema completo de la representación sino a uno de sus componentes, ese que caracterizó al racionalismo clásico: al modelo cartesiano del orden metódico. En consecuencia, podemos constatar la pervivencia del clasicismo actualmente. Las músicas populares, las dramaturgias cinematográficas, etc. van a promover un nuevo género que regresa a las fuentes populares, aunque se aleja de los patrones musicales del clasicismo ya convertido en música culta, música clásica. El mundo de la comedia clásica está en nuestra sensibilidad, las estructuras espectaculares creadas en aquella época siguen funcionando. Este mundo de la comedia ilustrada ha llegado a convertirse en la dramaturgia de la cultura de masas que la Dialéctica de la Ilustración llamó “industria cultural”. Este trabajo ha pretendido realizar la genealogía de los contenidos de esa cultura que impregna todos los productos culturales que se producen en la segunda mitad del siglo XX, especialmente después de que la ilusión de la crisis de la representación haya acabado con la Vanguardia. Y a través de ese análisis hemos definido un modelo crítico de la actitud ciudadana. Respecto a esa construcción cultural se construyen dos figuras del espectador: la ofrecida por Rousseau y otra de Diderot. Nuestra perspectiva es más diderotiana que roussoniana: el espectador debe dejar de serlo, pero en el teatro mismo, no contra el teatro. Nuestras formas culturales y espectaculares forman parte de nuestra identidad y, por ello, debemos exigir no solo su mantenimiento sino la calidad de su custodia, su renovación y su desarrollo. / The objective of this research is to set the semantic field of the concept of representation at the time of its origin (17th and 18th centuries). We aim to identify the various areas of the experience on which this conceptual structure is based upon and, at the same time, describe the mutations it has suffered up to its crisis, in its triple aspect: artistic, epistemological and juridical. We will try to show how the old metaphor of the Theatrum Mundi, freed from the theological system, became a form of spectacular society, during the Baroque period Our hypothesis is that the new concept of contractual political power, depends on the metaphor of the theatrical performance, which is the indisputable paradigm of this cultural model. The citizen becomes a spectator both at the theatre as in society and politics. New relations of power and new individuals (the bourgeoisie and the urban masses), but also new urban and civic spaces: real and open (squares and gardens), or virtual and closed (theatres). These aesthetic-political complex performances will reveal through the musical show invented in the 16th century: the opera. During the 18 th. century, the comic opera became a hallmark of the Enlightenment and the bourgeoisie. The texts have been submitted by a hermeneutics methodology by doing a transversal reading about all those original documents presenting a perspective of this scenic metaphor. The current issue of political representation has been treated since the genealogical method using three perspectives. The first one by presents the three disciplines which we know the representation in the West: political theory, epistemology and aesthetics. The second perspective, that illuminates a capital element within this system, makes reference to the representative characters: the politician, the scientist and the artist. Finally, the representation cultivated by these three characters is the performance that is offered to the public, which "acts" in the representation as the represented: the spectator (the crowd-audience), which is the third conceptual figure or perspective. The result of this process lets us see how, thanks to that form of social differentiation based on the distinction or aesthetic taste, opposite the opera and other artistic manifestations, a spectacular system absolutely vertical was configured. High cultures addressed those distinguished individuals converted into emblematic (representative) characters (gentry). We have discovered the keys to the understand the modern public man as a symbol, through our reflections about drama. The public space has been transformed into a stage (the great theatre of the world) and is presented in the form of representation. The society, the city, has become a stage for the show; the citizen is the public and for it proceeds to organize complex representation. The professional figure of the public affairs representative is designed: the political representative, almost in the same way that there is a protagonist of the representation in the Theatre: the actor. One of the most important conclusions is that performance has not entered into crisis, but strengthened enormously while the modern global network is organized by the media that manage public opinion and stimulate mass consumption. Therefore, the crisis has not affected the whole system of performance, but us one of its components, the one which characterized classical rationalism, the Cartesian model of the methodical order. As a result, we can recognize the survival of classicism: popular music, the cinema dramaturgy, etc. generate a new genre that returns to the popular sources, although it moves away from the musical patterns of classicism as converted into art music, classical music. The world of classical comedy is in our sensitivity, the spectacular structures created at that time are still operating. This world of the illustrated comedy has become the drama of mass culture that Dialectics of Enlightenment called cultural industry. This work has tried to carry out the genealogy of the contents of that culture that permeates all cultural products that are produced in the second half of the 20th century, especially since the illusion of the crisis of performance finished with the avant-garde. Through this analysis, we have defined a critical model of citizen attitude. Two figures of the viewer are built on that cultural construction: the one offered by Rousseau and another by Diderot. Our perspective is more Diderot-like than Roussonian: the spectator must cease to be, but within the theatre itself, not against the theatre. Our cultural and spectacular forms are part of our identity, thus we must demand not only its maintenance but also the quality of its custody, renewal and development.

Chromosomal anomaly is an abnormality of the number or structure of the chromosome. Based on this, they are classified as either numerical or structural. These anomalies can have a mild or severe effect on the phenotype of the carrier which depends on the chromosomal region involved and the genes implicated. They can be sporadic or inherited. Thus, it is essential to investigate such anomalies both prenatally and postnatally alike. In a standard cytogenetic lab, these anomalies can be detected using low-resolution or high-resolution karyotype, usually by performing GTG (G-bands by trypsin using Giemsa) on chromosomes undergoing mitosis, derived from blood. If higher resolution is required, there are various other molecular cytogenetic techniques available like FISH and microsatellite analysis. More recently, microarrays have opened a new era in the field of molecular genetics by greatly increasing the resolution of screening for copy number gains and losses. Using these techniques, we characterized deletions in the long arm of chromosome 7 in three clinical cases, identified the breakpoint, studied the inheritance pattern and compared our cases with the other cases carrying similar deletion in the literature. It was observed that the deletion was located at 7q36.2, 7q35 and 7q36.1 for cases 1, 2 and 3 respectively. Case 3 also carried a duplication of Xq28.

Este trabajo de investigación reconstruye y comenta la trayectoria artística de Silvia Gubern, Àngels Ribé y Eulàlia (Grau Donada), principalmente en la etapa de las llamadas “prácticas conceptuales”, a partir de fuentes primeras y documentales e introduciendo la voz y el relato de las artistas. Estas tres monografías incorporan también la recepción crítica y la valoración de la prensa especializada. La autora propone lecturas y reflexiones de estas prácticas artísticas y de la concepción de la creatividad de las artistas, vinculándolas a partir de las orientaciones que asume como guías teóricas del pensamiento de la diferencia sexual sobre la libertad de las mujeres en los 60 y 70. Mas allá de los marcos interpretativos “neutrales” que se ha utilizado para clasificar a estas artistas como conceptuales en la historia “modernista” del arte catalán y español, este trabajo de investigación pone las bases para leerlas a partir de genealogías de la creación y la creatividad orientadas por la sexuación del conocimiento en femenino. / This research builds and discusses the artistic career of Silvia Gubern, Àngels Ribé and Eulàlia (Grau), mainly in the stage called "conceptual practices" from documentary sources and introducing the voice of the artists and biographic facts. These three monographs also incorporate critical reception and evaluation of the press and the art critics. The author points out the importance of reading those careers taking into account the perspective of the Italian and Spanish feminism of sexual difference. Beyond the "neutral" interpretive framework that has been used to classify them as conceptual artists in the modernist history of catalan avantgarde art, this thesis lays the foundation for reading women genealogies of creativity oriented by the female sexuation of knowledge and to explore women's freedom in history. Thus, conceptualism would not be a single framework to assess their contribution to the history of art. It is defined as a living moment in a much longer and complex creative itinerary, a stage in which many issues intersect, focusing on some core aspects such as sexual politics in the history of women in the 60s and 70s in Catalonia (Spain).

Στον χώρο της Πανεπιστημιούπολης εντοπίστηκε η ύπαρξη γεωλογικής ασυνέχειας. Πρόκειται πιθανά για ρήγμα που ήδη έχει προκαλέσει διαφοροποίησεις σε κτίρια του πανεπιστημίου όσο καί στον δρόμο των αυτοκινήτων. Με στόχο την χαρτογράφηση της εν λόγω γεωλογικής ασυνέχειας εφαρμόστηκε λεπτομερής γεωφυσική διερεύνηση βασιζόμενη σε δύο περιοχές της Πανεπιστημιούπολης μέσω της ηλεκτρικής διασκόπησης. Σκοπός της έρευνας ήταν η καταγραφή της κατανομής της ηλεκτρικής καί ειδικής ηλεκτρικής αντίστασης του εδάφους σε διάφορα βάθη, ώστε να επιτευχθεί η λεπτομερής αποτύπωση της γεωλογικής ασυνέχειας. Πραγματοποιήθηκε διαχωρισμός της περιοχής σε γεωφυσικά δίκτυα, στα οποία εφαρμόστηκε ηλεκτρική χαρτογράφηση μέσω της δίδυμης διάταξης όσο καί κατακόρυφη ηλεκτρική απεικόνιση μέσω των διατάξεων Wenner & Wenner - Schlumberger. Η σάρωση των ανωτέρω γεωφυσικών δικτύων πραγματοποιήθηκε τόσο παράλληλα όσο καί κάθετα στον γεωμαγνητικό βορρά με άνοιγμα ηλεκτροδίων από 1 έως καί 3 μέτρα, με βήμα 0.5 έως 1 μ. Η αποτύπωση της χαρτογράφησης αποδόθηκε με την μορφή έγχρωμων χαρτών όπου παρατίθεται η κατανομή της μετρούμενης φυσικής ιδιότητας. Μέσω ειδικών λογισμικών γεωφυσικής επεξεργασίας ( Oasis Montaj, Res2dinv ) επιτεύχθηκε η απόδοση της ηλεκτρικής αντίστασης καί της ειδικής ηλεκτρικής αντίστασης. Οι χάρτες αυτοί απεικονίζουν την διασπορά των μετρούμενων φυσικών ιδιοτήτων του εδάφους τόσο επί παράλληλου όσο καί επί κατακόρυφου επιπέδου. Τέλος μέσω πίνακα αντιστάσεων από τη διεθνή βιβλιογραφία, έγινε προσπάθεια για την ερμηνεία των εμφανιζόμενων γεωφυσικών ανωμαλιών. / A detailed geophysical investigation applied on an region of University of Patras. Main objective of the process, was a detailed recording of an existing fault impact, by using combined geophysical techniques. The real activity of the above fault produced damages in human buildings. Especially the main road of University was destroyed by subtraction, while buildings at University students union, were abandoned due to huge subtractions episode. The detailed geophysical research applied on two separated geophysical grids, which were located about 100 m away each other. Firstly a geoelctric mapping technique took place, by using four electrodes in distance 0.5,1,2,3 meters and by adopting the twin-probe array arrangement. As result was the recording of soil resistance in a depth, on a horizontal plane. Secondly applied the geoeltric imaging technique. Both of geophysical grids were researched by using twenty-five equal space electrodes, which were located on a straight line in constant space. As result was the recording of distribution of resistivity soil on a vertical layer. The first and second techniques were applied parallel and perpendicular to the existing geomagnetic north axis. By further processing, the distribution of soil resistance and resistivity were illustrated on color map schedule by using the special geophysical processing software, known as Oasis Montaj. Also by using the special slice software ( Stephanopoulos , 2002 ) horizontal slices produced from parallel vertical section, where the distribution of resistivity was illustrated on constant depth.

Σκοπός της παρούσας εργασίας είναι ο σχεδιασµός η ανάπτυξη και η κατασκευή ενός καρδιογράφου ελεγχόµενου από υπολογιστή. Ο ηλεκροκαρδιογράφος είναι µια συσκευή η οποία µπορεί να συλλέγει από το ανθρώπινο σώµα και να απεικονίζει σε κάποιο µέσο τα δυναµικά που προκαλούνται από την δραστηριότητα της καρδιάς. Τα σήµατα αυτά, έχουν συγκεκριµένη µορφή και η απεικόνισή τους στον χρόνο µπορεί να µας δώσει πολύτιµες πληροφορίες σχετικά µε την κατάσταση της καρδιάς, βοηθώντας µας να εξάγουµε συµπεράσµατα σχετικά µε αυτή. / The aim of the present thesis is the design and development of a digital, computer controlled Cardiograph

Το ενδογενές σύστημα των κανναβινοειδών αποτελεί ένα πολύπλοκο ενδογενές σύστημα μεταγωγής σήματος το οποίο επηρεάζει ένα σημαντικό αριθμό φυσιολογικών διεργασιών και μεταβολικών μονοπατιών του οργανισμού (Cota and Woods, 2005). Απαρτίζεται από τους διαμεμβρανικούς υποδοχείς των κανναβινοειδών (CBR), τα ενδοκανναβινοειδή και τις πρωτεΐνες που είναι υπεύθυνες για την βιοσύνθεση και την αποικοδόμηση των δεύτερων. (Petrocellis et al., 2004). Τα κανναβινοειδή παρουσιάζουν ποικιλία επιπτώσεων, όπως δυσλειτουργία στη μάθηση και μνήμη, διαφοροποίηση των συναισθηματικών καταστάσεων, μείωση κινητικού ελέγχου και αναλγησία. Αναστέλλουν τη συναπτική διαβίβαση σε διάφορες περιοχές του εγκεφάλου όπως ο ιππόκαμπος, ο επικλινής πυρήνας και ο προμετωπιαίος φλοιός κυρίως μέσω προσυναπτικών μηχανισμών. Το γλουταμινικό οξύ είναι ο κύριος διεγερτικός νευροδιαβιβαστής στο Κεντρικό Νευρικό Σύστημα των θηλαστικών. Οι γλουταμινεργικοί νευρώνες διανέμονται ευρέως στο κεντρικό νευρικό σύστημα και παίζουν ρόλο σε πολλές βιολογικές διεργασίες, συμπεριλαμβανομένης της κωδικοποίησης των πληροφοριών, το σχηματισμό και την ανάκτηση των μνημών, τη χωρική αναγνώριση και τη διατήρηση της προσοχής (McEntee και Crook, 1993). Λόγω του ρόλου του στη συναπτική πλαστικότητα, το γλουταμινικό εμπλέκεται σε γνωσιακές λειτουργίες του εγκεφάλου, όπως η μάθηση και η μνήμη (McEntee και Crook, 1993). Το γλουταμινικό δρα στα μετασυναπτικά κύτταρα σε τρεις οικογένειες ιοντοτροπικών υποδοχέων τους NMDA, τους AMPA και τους καϊνικούς υποδοχείς, ενώ υπάρχουν, επίσης, και γλουταμινικοί μεταβολοτροπικοί υποδοχείς που συνδέονται με G πρωτεΐνες. Η αμυγδαλή εκτελεί ένα πρωταρχικό ρόλο στην επεξεργασία των συναισθηματικών αντιδράσεων και τη μνήμη, ενώ ο ιππόκαμπος έχει σημαντικό ρόλο στην εδραίωση των πληροφοριών από βραχυχρόνια μνήμη σε μακροχρόνια μνήμη και στη χωρική πλοήγηση. Οι δύο αυτές περιοχές είναι πλούσιες σε CB1 και NMDA υποδοχείς, ενώ τα τελευταία χρόνια, πληθώρα ερευνών υποδεικνύει ότι η έκθεση πειραματοζώων σε κανναβινοειδή επιφέρει σημαντικές αλλαγές σ’ αυτές τις δύο περιοχές. Στόχος της ερευνητικής εργασίας ήταν η μελέτη της επίδρασης των κανναβινοειδών στο γλουταμινεργικό σύστημα, στην αμυγδαλή και τον ιππόκαμπο επίμυων. Η μελέτη επικεντώθηκε στους NMDA υποδοχείς του γλουταμινικού οξέος με τη χρήση των μεθόδων της in situ υβριδοποίησης για το mRNA των NR1, NR2A και NR2B υπομονάδων του υποδοχέα και της ποσοτικής αυτοραδιογραφίας υποδοχέων, σε επίμυς που τους χορηγήθηκε WIN55212-2, ένας συνθετικός αγωνιστής του CB1 κανναβινοειδικού υποδοχέα, ο οποίος εμφανίζει παρόμοια δράση με την Δ9-τετραϋδροκανναβινόλη (THC), το φυσικό συστατικό του φυτού Cannabis sativa. Μελετήθηκαν τέσσερις ομάδες επίμυων όπου σε δύο από αυτές χορηγήθηκε WIN και στις υπόλοιπες δύο χορηγήθηκε ο διαλύτης του WIN, Vehicle. Στις δύο ομάδες που χορηγήθηκε WIN55212- 2 1mg/kg για 20 ημέρες, οι επίμυς της μίας ομάδας (ομάδα WIN) θανατώθηκαν 2 ώρες μετά από την τελευταία δόση ενώ οι επίμυς της άλλης ομάδας (ομάδα WIN+WITHD) θανατώθηκαν 7 ημέρες μετά την τελευταία δόση. Κατά το διάστημα αυτό δεν έγινε καμία χορήγηση ώστε να μελετηθεί αν οι επιπτώσεις της χρόνιας χορήγησης του WIN ήταν μόνιμες. Τα αποτελέσματά μας έδειξαν ότι μετά τη χρόνια χορήγηση του WIN τα επίπεδα έκφρασης των υπομονάδων του NMDA υποδοχέα μειώθηκαν στο ιππόκαμπο και την αμυγδαλή των επίμυων, ενώ μετά την διακοπή της χορήγησης του WIN, τα επίπεδα έκφρασης επανέχρονται. Είναι πιθανό ότι οι αλλαγές στην έκφραση και λειτουργία των υποδοχέων του γλουταμινικού που παρουσιάζονται ως προσαρμοστικές αλλαγές στο νευρικό σύστημα, να είναι ένας κοινός μηχανισμός με τον οποίο τα εθιστικά ναρκωτικά επηρεάζουν την νευρική λειτουργία. / The endocannabinoid system is a complicated endogenous signaling system that affects a variety of physiological processes and metabolic routes in human body (Cota and Woods, 2005). The endocannabinoid system includes the transmembrane cannabinoid receptors (CBR), the endocannabinoids and the enzymes that synthesize and degrade the endocannabinoids (Petrocellis et al., 2004). Cannabinoids have a variety of effects, such as impairment in learning and memory, modulation of emotional states, reduced motor control and analgesia. Cannabinoids inhibit synaptic transmission in several brain regions such as the hippocampus, nucleus accumbens and the prefrontal cortex mainly via presynaptic mechanisms. Glutamate is the most abundant excitatory neurotransmitter in the mammalian central nervous system. The glutamatergic neurons are widely distributed in the central nervous system and play a role in many biological processes, including the coding of information, the formation and recovery of memories, the spatial recognition and maintaining the attention (McEntee και Crook, 1993). Because of its role in synaptic plasticity, glutamate is involved in cognitive functions like learning and memory in the brain. (McEntee και Crook, 1993). Glutamate activates three families of ionotropic receptors in postsynaptic cells, those are NMDA, AMPA and kainate receptors, while there are also metabotropic G proteins coupled glutamate receptors. Amygdala has a primary role in the processing of emotional reactions and memory, whereas hippocampus has an important role in the consolidation of informations from short term memory into long term memory and spatial navigation. These two brain regions contain a large number of CB1 and NMDA receptors, while recently, many studies suggest that animals treated with cannabinoids display significant changes in these two areas. The aim of this research was to study the changes in glutamatergic system in the amygdala and hippocampus of rats treated with cannabinoids. We focused on NMDA glutamate receptors, using in situ hybridization for studying the expression of NR1, NR2A and NR2B subunits and quantitative receptor autoradiography, in rats treated with WIN55212-2, a synthetic agonist of the CB1 cannabinoid receptor, which shows similar effects with delta-9-tetrahydrocannabinol (THC), a natural component of the plant Cannabis sativa. Adult rats were injected with WIN55212-2 (1mg/kg) and Vehicle. Animals received repeated administrations of WIN55212-2 1mg/kg once a day for 20 days. Animals in group WIN were sacrificed 2 hours after the last administration whereas in group WIN + WITHD were sacrificed 7 days after the last administration. During this time there was no administration of WIN so we could study whether the effects of chronic exposure were permanent. Our results demonstrate that chronic exposure to WIN55212-2 produced significant decreases in the expression of NMDA receptor's subunits in hippocampus and amygdala. These changes were reversed one week after abstinence. These adaptive synaptic changes may share common mechanisms with addictive drugs in modifying neural circuitry.