Esofagite a citomegalovirus em cirrose hepática auto-imune : a propósito de um caso clínico

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

FAULT MORPHOLOGY WITHIN THE SOUTHERN KENYAN PORTION OF THE EAST AFRICAN RIFT VALLEY

Faults form quickly, geologically speaking, with sharp, crisp step-like profiles. Logic dictates that erosion wears away this "sharpness" or angularity creating more rounded features. As erosion occurs, debris accumulates at the base of the scarp slope. The stable end point of this process is when the scarp slope approaches an ideal sigmoid shape. This theory of fault end process, in combination with a new method developed in this report for fault profile delineation, has the potential to enable observation and categorization of fault profiles over large, diverse swaths of fault formation-- in remote areas such as the Southern Kenyan Rift Valley. This up-to date method uses remote sensing data and the digitizer tool in Global Mapper to create shape files of fault segments. This method can provide further evidence to support the notion that sigmoidal- shaped profiles represent a natural endpoint of the erosional process of fault scarps. Over time, faults of many different ages would exist in this similar shape over a wide region. However, keeping in mind that other processes can be at work on scarps-- most notably drainage patterns, when anomalies in profiles are observed, reactivation in some form possibly has occurred.

Single-nucleotide polymorphism in two representative multidrug-resistant Mycobacterium bovis isolates collected from patients in a Spanish hospital harboring a human infection outbreak

Mycobacterium bovis is the etiological agent of tuberculosis in domestic and wild animals. Its involvement as a human pathogen has been highlighted again with the recent descriptions of transmission through dairy products (18), reactivation or primary infection in human immunodeficiency virus-infected patients (5), and association with meat industry workers, animal keepers, or hunters (3). Strains resistant to antituberculous drugs (M. bovis is naturally resistant to pyrazinamide) pose an additional risk (2). Several studies have demonstrated that mutations in target genes are associated with resistance to antituberculous drugs (4, 7, 10, 11, 16). However, most of them have been developed in Mycobacterium tuberculosis strains and limited data are available regarding M. bovis isolates. The aim of this study was to characterize by sequencing the main genes involved in antibiotic resistance in two multidrug-resistant (MDR) M. bovis isolates in a human outbreak detected in a hospital in Madrid that subsequently spread to several countries (5, 6, 15). The isolates were resistant to 11 drugs, but only their rpoB and katG genes have been analyzed so far (1, 14). We studied the first (93/R1) and last (95/R4) M. bovis isolates of this nosocomial outbreak, characterized by spoligotyping as SB0426 (hexacode 63-5F-5E-7F-FF-60 in the database at www.mbovis.org) (1, 13). Several genes involved in resistance to isoniazid (katG, ahpC, inhA, and the oxyR-ahpC intergenic region), rifampin (rpoB), streptomycin (rrs, rpsL), ethambutol (embB), and quinolones (gyrA) were studied. These genes, or fragments of genes, were amplified and sequenced as previously described (12). The sequence analysis revealed polymorphisms in five (ahpC, rpoB, rpsL, embB, and gyrA) out of nine analyzed genes (Table 1). Nucleotide substitutions in four genes cause a change in the encoded amino acid. Two additional synonymous mutations in ahpC and rpsL differentiated the first and last isolates from the outbreak.

Evaluación del desempeño de pruebas diagnósticas de tuberculosis en pacientes VIH positivo en dos hospitales públicos de Bogotá

La tuberculosis TB es una de las principales causas de muerte en el mundo en individuos con infección por VIH. En Colombia esta coinfección soporta una carga importante en la población general convirtiéndose en un problema de salud pública. En estos pacientes las pruebas diagnósticas tienen sensibilidad inferior y la enfermedad evoluciona con mayor frecuencia hacia formas diseminadas y rápidamente progresivas y su diagnóstico oportuno representa un reto en Salud. El objetivo de este proyecto es evaluar el desempeño de las pruebas diagnósticas convencionales y moleculares, para la detección de TB latente y activa pacientes con VIH, en dos hospitales públicos de Bogotá. Para TB latente se evaluó la concordancia entre las pruebas QuantiFERON-TB (QTF) y Tuberculina (PPD), sugiriendo superioridad del QTF sobre la PPD. Se evaluaron tres pruebas diagnósticas por su sensibilidad y especificidad, baciloscopia (BK), GenoType®MTBDR plus (Genotype) y PCR IS6110 teniendo como estándar de oro el cultivo. Los resultados de sensibilidad (S) y especificidad (E) de cada prueba con una prevalencia del 19,4 % de TB pulmonar y extrapulmonar en los pacientes que participaron del estudio fue: BK S: 64% E: 99,1%; Genotype S: 77,8% E: 94,5%; PCRIS6110 S: 73% E: 95,5%, de la misma forma se determinaron los valores predictivos positivos y negativos (VPP y VPN) BK: 88,9% y 94,8%, Genotype S: 77,8% E: 94,5%; PCRIS6110 S: 90% y 95,7%. Se concluyó bajo análisis de curva ROC que las pruebas muestran un rendimiento diagnóstico similar por separado en el diagnóstico de TB en pacientes con VIH, aumentando su rendimiento diagnostico cuando se combinan

Incidencia de la gestión de Frontex y el gobierno italiano en la acentuación de la crisis humanitaria de la Isla de Lampedusa entre 2011 y 2015

El presente estudio de caso busca examinar la incidencia de las medidas migratorias de control fronterizo implementadas por el Frontex y el gobierno Italiano en las condiciones mínimas de supervivencia de los migrantes irregulares, económicos y solicitantes de asilo en la Isla de Lampedusa, en el periodo 2011-2015. De esta manera, se identifican las medidas migratorias de control fronterizo implementadas por Frontex y el gobierno Italiano. Se examina la situación de la seguridad humana en la crisis migratoria de la Isla, y se analiza la relación entre las medidas migratorias de control fronterizo y las condiciones mínimas de supervivencia de los migrantes. El resultado de la investigación permite plasmar, las consecuencias negativas que han tenido las medidas migratorias en cuanto a las condiciones mínimas de supervivencia, lo que ha desembocado en una crisis humanitaria.

Lugares de montanha: proposta de reactivação de uma aldeia do Vale do rio Ceira, Serra da Lousã

O território da Serra da Lousã, quase desabitado e profundamente descaracterizado, esconde lugares em ruína e indícios de vivências passadas que a montanha teima em reclamar. Importa recuperar esses lugares reinventando-os, sob pena de se perderem com o passar do tempo e o avanço do meio natural. Este trabalho debruça-se sobre um dos lugares abandonados desta serra, a Silveira de Baixo, procurando nas suas raízes oportunidades para uma reactivação. Estuda a hipótese de tornar a aldeia e esta parte da serra, acessíveis a pessoas portadoras de deficiências, físicas ou cognitivas, tendo assim um duplo objetivo: permitir o acesso para todos a lugares de grande beleza natural e reactivar uma economia local que permita um desenvolvimento sustentado, através de actividades e produtos nativos deste território, conservando assim alguns dos seus saberes ancestrais. Desenha-se deste modo o início de uma comunidade, experimentando um equilíbrio entre o antigo e o contemporâneo, negando a musealização do lugar que o pode tornar estéril e constitui um impedimento no retorno de vida a este território. Assim, o trabalho experimenta o papel da arquitectura na reactivação de um lugar antigo e com marcas, preservando os seus elementos notáveis e com eles construindo novas espacialidades que ajudem ao estabelecimento de vivências contemporâneas; Mountain setlements: Proposal for reactivating an abandoned village in Ceira river valley, Lousã mountain range Abstract: The Lousã mountain range, almost uninhabited and profoundly decharacterized, speaks to us through its ruins about ways of living that ceased to exist. It is important to recover and reinvent them, otherwise they will slowly vanish as time goes by and nature steps in. This investigation focus on one of these abandoned settlements, Silveira de Baixo. It looks in its roots for opportunities of reactivation, investigating the hypothesis of turning this village and the mountain accessible to the physically and mentally impaired, thus assuming a double objective, of allowing these disabled people to experience a place of extreme beauty and of reinventing and reactivating native productive activities thus conserving them and allowing for the village’s economical sustainability. This work designs a new beginning for a community and experiments the balance between the old and the new, without excessive patrimonialisation that often kills the return of life to such settlements. Experimenting the role of architecture in providing life to an abandoned place through a sustainable intervention.

Les machines du pouvoir. Technique et politique entre l'Iran sassanide et le califat abbasside

Dans l’Antiquité, la recherche sur la technique permet les premières réalisations de dispositifs ingénieux, tels que des appareils qui accomplissent une série d’actions par le biais de stimulus externes et de mécanismes cachés. Les organismes politiques et religieux saisissent rapidement la puissance communicative de ces machines, en devenant les promoteurs et patrons privilégiés de leur production. L’Empire sassanide (224-650) ne constitue pas une exception. En effet, les souverains perses consacrent, au moins à l’époque tardive, une grande attention à la conception et au déploiement de dispositifs savants. De même, un siècle plus tard, dans le milieu du califat islamique, les Abbassides (750-1258) semblent s’entourer de tels dispositifs. La continuité entre les deux empires dans plusieurs domaines, de la théorie politique à l’administration, est bien connue. Cependant, la question de la réutilisation du patrimoine technique et scientifique ancien, et notamment sassanide, par la cour abbasside, demeure encore largement inexplorée. L’étude d’un corpus de sources, aussi vaste qu’hétérogène, rassemblant des ouvrages historiographiques, géographiques, poétiques et d’adab, ainsi que des traités scientifiques et techniques en plusieurs langues, permet d’analyser différents aspects de la production et de l’usage politique des machines. Au sein de la cour sassanide, comme de la cour abbasside, la machine s’avère constituer un véhicule préférentiel de représentation et de diffusion de l’idéologie politique. À travers sa mise en scène publique, elle contribue de manière substantielle à la définition de l’espace du pouvoir, en participant à la création d’une image de la cour comme un microcosme au cœur duquel le Roi des rois, et plus tard le calife, occupaient le rôle cardinal de maître incontesté du monde. La continuité entre les empires sassanide et abbasside dans le domaine technique ne se limite donc pas à une récupération de savoirs, mais s’opère aussi sous la forme d’une véritable réactivation d’un patrimoine symbolique

Preclinical studies of the combined effect of anti-MYCN PNA and Retinoic Acid as potential approach to treat Neuroblastoma

Neuroblastoma (NB) is the deadliest cancer in early childhood. Around 25% of patients pre- sent MYCN-amplification (MNA) which is linked to poor prognosis, metastasis, and therapy- resistance. While retinoic acid (RA) is beneficial only for some NB patients, the cause of its resistance is still unknown. Thus, there remains a need for new therapies to treat NB. I show that MYCN-specific inhibition by the antigene oligonucleotide BGA002 in combination with 13-cis RA (BGA002-RA) overcome resistance in MNA-NB cell lines, leading to potent MYCN mRNA expression and protein decrease. Moreover, BGA002-RA reactivated neuron differentiation or led to apoptosis in MNA-NB cell lines, and inhibited invasiveness capacity. Since NB and PI3K/mTOR pathway are strictly related MYCN down-regulation by BGA002 led to mTOR pathway inhibition in MNA-NB, that was strengthened by BGA002-RA. I further analyzed if MYCN silencing may induce autophagy reactivation, and indeed BGA002-RA caused a massive increase in lysosomes and macrovacuoles in MNA-NB cells. In addition, while MYCN is known to induce angiogenesis, BGA002-RA in vivo treatment elim- inated the tumor vascularization in a MNA-NB mice model, and significantly increased the survival. Overall, these results indicate that MYCN modulation mediates the therapeutic efficacy of RA and the development of RA resistance in MNA-NB. Furthermore, by targeting MYCN, we show a cancer-specific way of mTOR pathway inhibition only in MNA-NB, avoiding side effects of targeting mTOR in normal cells. These findings warrant clinical testing of BGA002-RA as a potential strategy to overcome RA resistance in MNA-NB.

Identificazione di un nuovo meccanismo microRNA-dipendente di resistenza all'inibizione della via di segnale PI3K/AKT nel carcinoma della prostata

Benché le alterazioni della via PI3K/AKT siano molto sudiate a causa del loro ruolo nella tumorigenesi, e rappresentino pertanto un importante bersaglio terapeutico, i risultati di numerosi studi clinici con inibitori di PI3K o AKT sono finora deludenti, in parte a causa dell’insorgenza di resistenza provocata dall'interruzione dei circuiti di feedback negativo. In questo studio, abbiamo scoperto che l’inattivazione farmacologica di AKT in cellule di carcinoma prostatico PC3 porta alla down-regolazione di un microRNA con funzione di oncosoppressore, il miR-145-5p, e ad un drammatico aumento di espressione di uno dei suoi geni target, cioè N/KRas. E’ interessante sottolineare che questo microRNA è considerato un marker di progressione metastatica nel carcinoma prostatico, il cui livello di espressione aiuta a discriminare tra pazienti con iperplasia prostatica benigna e cancro alla prostata. Inoltre, la bassa espressione di miR-145 aumenta il rischio di progressione della malattia da localizzata a metastatica. La conferma che l’aumento di Ras, osservato sia in termini di mRNA che di proteina, è dipendente dalla caduta del miR-145-5p, è stata poi ottenuta tramite un modello di PC3 ingegnerizzate per ottenere il silenziamento inducibile del miR-145-5p. Tramite un array di fosfoproteine siamo poi stati in grado di verificare che l’aumento di Ras provoca la riattivazione della cascata di PI3K/AKT e di ERK. Dal punto di vista meccanicistico, quindi, lo studio ha portato all’identificazione di un nuovo meccanismo di resistenza adattativa, in cui l’inattivazione di AKT provoca una caduta del miR-145-5p che, a sua volta, aumenta l’espressione di Ras e riattiva il signaling di PI3K, rendendo inefficace il trattamento farmacologico. Questi risultati sono particolarmente rilevanti alla luce di recenti studi (NCT04493853; NCT03072238; NCT02525068) e di trial clinici in corso (NCT04737109; NCT03673787), basati sulla somministrazione combinata di inibitori della sintesi degli androgeni con gli inibitori di AKT capitasertib o ipatasertib.

Preclinical evaluation of a MYCN-specific inhibitor for the treatment of small cell lung cancer

Small cell lung cancer (SCLC) is the most aggressive form of lung cancer, characterized by rapid growth, early metastasis and acquired drug resistance. SCLC is usually sensitive to initial treatment, however, most patients relapse within few months; thus more effective therapies are urgently needed. Key genetic alterations very frequently observed in SCLC include loss of TP53 and RB1 and mutations in the MYC family genes (MYC, MYCL or MYCN). One of them is amplified and overexpressed in a mutually exclusive manner and represents the most prominent activating oncogene alteration in this malignancy. In particular, MYCN amplification is associated with tumor progression, treatment failure and poor prognosis. Given the role of MYCN in SCLC and its restricted expression profile, MYCN represents a promising therapeutic target; although it is considered undruggable by traditional approaches. An innovative approach to target the oncogene concerns specific MYCN expression inhibition, acting directly at the level of DNA, through an antigene peptide nucleic acid (agPNA) oligonucleotide, called BGA002. This thesis focused on the study of BGA002, as a possible targeted therapeutic strategy for the treatment of MYCN-related SCLC. In this context, BGA002 proved to be a specific and highly effective inhibitor. Furthermore, MYCN silencing induced alterations in many downstream pathways and led to apoptosis, in concomitance with autophagy reactivation. Moreover, systemic administration of BGA002 was effective in vivo as well, significantly increasing survival in MNA mouse models, even in the scenario of multidrug-resistance. In addition, BGA002 treatment successfully reduced N-Myc protein expression and, more importantly, caused a massive diminishment in tumor vascularization in the multidrug-resistant model. Overall, these results proved that MYCN inhibition by BGA002 may represent a new promising precision medicine approach, to treat MYCN-related SCLC.