Diversidade genética e resistência natural ao Maraviroc em estirpes do vírus da imunodeficiência humana Tipo 1 (HIV-1) em circulação em utilizadores de drogas por via endovenosa na Grande Lisboa

RESUMO: O maraviroc (MVC) é o único anti-retroviral antagonista do co-receptor CCR5 licenciado e interage com as ansas transmembranares de CCR5, induzindo uma alteração da sua conformação e impedindo a interacção com gp120. O MVC é activo apenas contra estirpes R5 de HIV-1, sendo utilizado em terapia de recurso. Neste trabalho, foi estudada a diversidade genética da região C2V3C3 do gene env de estirpes de HIV-1 de oxicodependentes por via endovenosa da Grande Lisboa, pesquisando-se também a presença de polimorfismos genéticos naturais. Foram utilizadas 52 amostras de plasma e para 35 destas foi amplificado por RT-nested PCR um produto de 565 pb. A análise filogenética revelou a seguinte distribuição de genótipos: 23 B (incluindo, provavelmente, 2 CRF14_BG), 8 A, 3 G e 1 F1. Após tradução, e por comparação com a sequência consenso B, verificou-se uma elevada frequência de polimorfismos genéticos, sendo encontradas algumas “assinaturas de aminoácidos” relativas aos subtipos não-B. Realizou-se ainda uma pesquisa de locais de N-glicosilação e a previsão da utilização de co-receptores (abordagem genotípica), com recurso às regras 11/25 e da carga líquida da ansa V3 e aos programas PSSM e geno2pheno[coreceptor]. Observou-se uma conservação genérica do número de locais de N-glicosilação e foram identificadas 5 sequências com tropismo X4 ou duplo. Por fim, com base na literatura, realizou-se uma pesquisa de polimorfismos genéticos associados a resistência ao MVC presentes na ansa V3. Foi observado um número elevado destas mutações. A presença dos padrões 11S+26V e 20F+25D+26V, num total de 3 sequências, é relevante, visto estes estarem inequivocamente associados à resistência in vivo ao MVC. Apesar de não estar ainda definido um perfil de resistência para o MVC, a presença das mutações encontradas, em indivíduos sem contacto prévio com o fármaco, trará implicações relevantes na sua gestão clínica, considerando a introdução do MVC na terapia de recurso.---------- ABSTRACT: Maraviroc (MVC) is the only CCR5 inhibitor licensed today. This drug interacts with the transmembrane helices of CCR5 co-receptor, inducing a conformation change of its extracellular loops and preventing the interaction with gp120. MVC is only active against R5 strains of HIV-1 and is currently used in salvage therapy. The genetic diversity of the env C2V3C3 region of HIV-1 strains from injecting drug users in the Greater Lisbon was studied, along with the presence of natural genetic polymorphisms. 52 plasma samples were used and the amplification by RT-nested PCR of a 565 bp-product was possible in 35 of them. The phylogenetic analysis revealed 23 sequences classified as subtype B (probably including 2 CRF14_BG), 8 A, 3 G and 1 F1. After translation, the presence of natural genetic polymorphisms was studied by comparison to a subtype B consensus. A high frequency of genetic polymorphisms was observed and significant “amino acid signatures” were found in association with non-B subtypes. A full characterization of the N-glycosylation sites was also performed and a coreceptor prediction (genotypic approach) was accomplished using the 11/25 and the V3 net charge rules and the programs PSSM and geno2pheno[coreceptor]. The number of N-glycosylation sites was generically preserved. Five sequences were defined as X4 or dual-tropic. Based on published data, a search for genetic polymorphisms, present in V3loop, associated to MVC resistance was finally undertaken. Several of such mutations were observed, being particularly interesting the presence of the patterns 11S+26V and 20F+25D+26V, in a total of 3 sequences, since these patterns have unequivocally been associated with MVC resistance in vivo. Although a resistance profile for MVC is not yet defined, the presence of these mutations in MVC-naïve populations may have significant impact in their clinical management in the future, especially considering the introduction of this drug in salvage therapy.

An analysis of teachers' environmental concerns: use of a conceptual change model

Tese de doutoramento em Ciências da Educação

Vulnerability of mediterranean watersheds to climate change

Desertification is a critical issue for Mediterranean drylands. Climate change is expected to aggravate its extension and severity by reinforcing the biophysical driving forces behind desertification processes: hydrology, vegetation cover and soil erosion. The main objective of this thesis is to assess the vulnerability of Mediterranean watersheds to climate change, by estimating impacts on desertification drivers and the watersheds’ resilience to them. To achieve this objective, a modeling framework capable of analyzing the processes linking climate and the main drivers is developed. The framework couples different models adapted to different spatial and temporal scales. A new model for the event scale is developed, the MEFIDIS model, with a focus on the particular processes governing Mediterranean watersheds. Model results are compared with desertification thresholds to estimate resilience. This methodology is applied to two contrasting study areas: the Guadiana and the Tejo, which currently present a semi-arid and humid climate. The main conclusions taken from this work can be summarized as follows: • hydrological processes show a high sensitivity to climate change, leading to a significant decrease in runoff and an increase in temporal variability; • vegetation processes appear to be less sensitive, with negative impacts for agricultural species and forests, and positive impacts for Mediterranean species; • changes to soil erosion processes appear to depend on the balance between changes to surface runoff and vegetation cover, itself governed by relationship between changes to temperature and rainfall; • as the magnitude of changes to climate increases, desertification thresholds are surpassed in a sequential way, starting with the watersheds’ ability to sustain current water demands and followed by the vegetation support capacity; • the most important thresholds appear to be a temperature increase of +3.5 to +4.5 ºC and a rainfall decrease of -10 to -20 %; • rainfall changes beyond this threshold could lead to severe water stress occurring even if current water uses are moderated, with droughts occurring in 1 out of 4 years; • temperature changes beyond this threshold could lead to a decrease in agricultural yield accompanied by an increase in soil erosion for croplands; • combined changes of temperature and rainfall beyond the thresholds could shift both systems towards a more arid state, leading to severe water stresses and significant changes to the support capacity for current agriculture and natural vegetation in both study areas.

Change Processes and Future Perspectives in the Knowledge Society. The Example of Clothing and Textile Industry

The paper examines change processes und future perspectives in the knowledge society. It presents the clothing and textile industry as an example for a transforming industry in a global economy. The paper reviews existing future studies, which have surveyed change processes and future developments in the clothing and textile industry. Main goals of the review are the identification of changes in work and the description of the restructuring of global value chains within the clothing and textile sector. The paper also highlights major current trends, drivers of change and future prospects in this sector.

Climate change impacts on portuguese energy system in 2050

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia do Ambiente, perfil Gestão e Sistemas Ambientais

Assessment of climate change statistical downscaling methods: Application and comparison of two statistical methods to a single site in Lisbon

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia do Ambiente

Land use / land cover change detection: an object oriented approach, Münster, Germany

Dissertation submitted in partial fulfilment of the requirements for the Degree of Master of Science in Geospatial Technologies

Urban land use change analysis and modelling: a case study of Setubal-Sesimbra, Portugal

Dissertation submitted in partial fulfilment of the requirements for the Degree of Master of Science in Geospatial Technologies

Urban change monitoring using GIS and remote sensing tools in Kathmandu valley (Nepal)

Dissertation submitted in partial fulfilment of the requirements for the Degree of Master of Science in Geospatial Technologies

Land use/ land cover change and its impact on soil erosion process in Begnas Tal Rupa Tal watershed using geospatial tools, Kaski district, Nepal

Dissertation submitted in partial fulfilment of the requirements for the Degree of Master of Science in Geospatial Technologies

Human and climatic change impact modelling on the habitat suitability for the chimpanzee (Pan troglodytes ellioti) – Case study: The proposed Mount Cameroon National Park

Dissertation submitted in partial fulfilment of the requirements for the Degree of Master of Science in Geospatial Technologies

Ground Vibrations Induced by High-Speed Trains in Regions with Sudden Foundation Stiffness Change

In this paper analytical transient solutions of dynamic response of one-dimensional systems with sudden change of foundation stiffness are derived. In more details, cantilever dynamic response, expressed in terms of vertical displacement, is extended to account for elastic foundation and then two cantilever solutions, corresponding to beams clamped on left and right hand side, with different value of Winkler constant are connected together by continuity conditions. The internal forces, as the unknowns, can be introduced by the same values in both clamped beam solutions and solved. Assumption about time variation of internal forces at the section of discontinuity must be adopted and originally analytical solution will have to include numerical procedure.

Thermodynamic and kinetic modelling of the redox properties of tetrahaem cytochromes C3

Dissertação apresentada para obtenção do grau de Doutor em Bioquímica,especialidade Bioquímica-Física, pela Universidade Nova de Lisboa, Faculdade de Cincias e Tecnologia

Spatial metrics and Landsat data for urban landuse change detection: case of Addis Ababa, Ethiopia.

Dissertation submitted in partial fulfilment of the requirements for the Degree of Master of Science in Geospatial Technologies.

Interpreting change in human-nature and long term social relationships

Dissertação apresentada para obtenção do Grau de Doutor em Ciências do Ambiente, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia.