Efeitos da acidificação da água do mar na reprodução de Mytilus edulis

Dissertação de mest., Biologia Marinha (Ecologia e Biodiversidade Marinha), Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

A integração dos assentamentos rurais tradicionais nas regiões urbanas: o exemplo do Algarve Central no Sul de Portugal

Versão em português do artigo submetido com o título “Towards Integrating Rural Vernacular Settlements in Urban Regions: A study of Algarve, Portugal” ao ISVS e-­‐journal em Abril de 2011.

Contributos operários para a educação popular em Portugal. O caso da Fábrica de Conservas S. Francisco, da empresa Júdice Fialho, em Portimão

Este artigo debruça-se sobre o período histórico do pós revolução de abril de 1974, a abril de 1976, usando como quadro concetual a noção de educação popular e como contexto empírico a fábrica de conservas “S. Francisco” da empresa Júdice Fialho, localizada na cidade de Portimão. Pretende abrir algumas ideias para perceber de que forma o operariado se organizou em comissões de base, para lutar em defesa de melhores condições de vida e de trabalho. Pretende, ainda, compreender os mecanismos usados para desenvolver um processo educativo de aprendizagem autónoma e emancipatória relativa ao trabalho e à vida social.

A importância da supervisão na eficácia da educação ambiental em contexto escolar

Dissertação de mest., Supervisão, Faculdade de Ciências Humanas e Sociais, Univ. do Algarve, 2008

Tavira, o Marquês de Pombal e a fábrica de tapeçarias

Trata-se de um trabalho destinado a revelar a existência no Algarve, mais propriamente em Tavira, de uma importante unidade fabril destinada a produzir tapeçarias, segundo a técnica francesa aplicada em Aubusson. São aqui também reveladas, com recurso a imagens e documentos de arquivo, dados a público em primeira mão, as primeiras tapeçarias fabricadas em Tavira, assim como as razões que forçaram o encerramento desta notável unidade fabril.

Formas de autorrepresentação e mutações contemporâneas da literacia

Desde 2008/ 2009 tenho lecionado a disciplina de licenciatura Autobiografia e Histórias de Vida no âmbito da qual pedi aos alunos que elaborassem um bilhete de identidade personalizado. Com este material pude desenvolver uma reflexão a partir do diálogo entre a forma por excelência de identificação pública em Portugal e o discurso identitário de jovens estudantes universitários. O processamento textual dos seus bilhetes de identidade revelou a estreita ligação com os suportes mediáticos que os enformam (na esmagadora maioria digitais) e com as práticas de literacia a eles associadas. Afinal de contas, estes estudantes vivem imersos num contexto de revolução digital em que as noções estabelecidas de literacia e os modos tradicionais de leitura e autorrepresentação estão a ser desafiados e até postos em causa.

An integrated and comparative approach to genetic selection of the aquaculture species Dicentrarchus labrax

The European sea bass, Dicentrarchus labrax, is one of the most important marine species cultivated in Southern Europe and has not benefited from selective breeding. One of the major goals in the sea bass (D. labrax) aquaculture industry is to understand and control the complexity of growth associated traits. The aim of the methodology developed for the studies reported in the thesis was not only to establish genetic and genomic resources for sea bass, but to also develop a conceptual strategy to efficiently create knowledge in a research environment that can easily be transferred to the aquaculture industry. The strategy involved; i) establishing an annotated sea bass transcriptome and then using it to, ii) identify new genetic markers for target QTL regions so that, iii) new QTL analysis could be performed and marker based resolution of the DNA regions of interest increased, and then iv) to merge the linkage map and the physical map in order to map the QTL confidence intervals to the sea bass genome and identify genes underlying the targeted traits. Finally to test if genes in the QTL regions that are candidates for divergent growth phenotypes have modified patterns of transcription that reflects the modified whole organism physiology SuperSAGE-SOLiD4 gene expression was used with sea bass with high growth heterogeneity. The SuperSAGE contributed to significantly increase the transcriptome information for sea bass muscle, brain and liver and also led to the identification of putative candidate genes lying in the genomic region of growth related QTL. Lastly all differentially expressed transcripts in brain, liver and muscle of the European sea bass with divergent specific growth rates were mapped to gene pathways and networks and the regulatory pathways most affected identified and established the tissue specific changes underlying the divergent SGR. Owing to the importance of European sea bass to Mediterranean aquaculture and the developed genomics resources from the present thesis and from other studies it should be possible to implement genetic selection programs using marker assisted selection.

Development of a Piggybac based direct reprogramming system for derivation of integration free induced pluripotent stem cells

Induced pluripotent stem cells (iPSc) have great potential for applications in regenerative medicine, disease modeling and basic research. Several methods have been developed for their derivation. The original method of Takahashi and Yamanaka involved the use of retroviral vectors which result in insertional mutagenesis, presence in the genome of potential oncogenes and effects of residual transgene expression on differentiation bias of each particular iPSc line. Other methods have been developed, using different viral vectors (adenovirus and Sendai virus), transient plasmid transfection, mRNA transduction, protein transduction and use of small molecules. However, these methods suffer from low efficiencies; can be extremely labor intensive, or both. An additional method makes use of the piggybac transposon, which has the advantage of inserting its payload into the host genome and being perfectly excised upon re-expression of the transposon transposase. Briefly, a policistronic cassette expressing Oct4, Sox2, Klf4 and C-Myc flanked by piggybac terminal repeats is delivered to the cells along with a plasmid transiently expressing piggybac transposase. Once reprogramming occurs, the cells are re-transfected with transposase and subclones free of tranposon integrations screened for. The procedure is therefore very labor intensive, requiring multiple manipulations and successive rounds of cloning and screening. The original method for reprogramming with the the PiggyBac transposon was created by Woltjen et al in 2009 (schematized here) and describes a process with which it is possible to obtain insert-free iPSc. Insert-free iPSc enables the establishment of better cellular models of iPS and adds a new level of security to the use of these cells in regenerative medicine. Due to the fact that it was based on several low efficiency steps, the overall efficiency of the method is very low (<1%). Moreover, the stochastic transfection, integration, excision and the inexistence of an active way of selection leaves this method in need of extensive characterization and screening of the final clones. In this work we aime to develop a non-integrative iPSc derivation system in which integration and excision of the transgenes can be controlled by simple media manipulations, avoiding labor intensive and potentially mutagenic procedures. To reach our goal we developed a two vector system which is simultaneously delivered to original population of fibroblasts. The first vector, Remo I, carries the reprogramming cassette and GFP under the regulation of a constitutive promoter (CAG). The second vector, Eneas, carries the piggybac transposase associated with an estrogen receptor fragment (ERT2), regulated in a TET-OFF fashion, and its equivalent reverse trans-activator associated with a positive-negative selection cassette under a constitutive promoter. We tested its functionality in HEK 293T cells. The protocol is divided in two the following steps: 1) Obtaining acceptable transfection efficiency into human fibroblasts. 2) Testing the functionality of the construct 3) Determining the ideal concentration of DOX for repressing mPB-ERT2 expression 4) Determining the ideal concentration of TM for transposition into the genome 5) Determining the ideal Windows of no DOX/TM pulse for transposition into the genome 6) 3, 4 and 5) for transposition out of the genome 7) Determination of the ideal concentration of GCV for negative selection We successfully demonstrated that ENEAS behaved as expected in terms of DOX regulation of the expression of mPB-ERT2. We also demonstrated that by delivering the plasmid into 293T HEK cells and manipulating the levels of DOX and TM in the medium, we could obtain puromycin resistant lines. The number of puromycin resistant colonies obtained was significantly higher when DOX as absent, suggesting that the colonies resulted from transposition events. Presence of TM added an extra layer of regulation, albeit weaker. Our PCR analysis, while not a clean as would be desired, suggested that transposition was indeed occurring, although a background level of random integration could not be ruled out. Finally, our attempt to determine whether we could use GVC to select clones that had successfully mobilized PB out of the genome was unsuccessful. Unexpectedly, 293T HEK cells that had been transfected with ENEAS and selected for puromycin resistance were insensitive to GCV.

O sítio da Ponte da Azambuja 2 (Portel, Évora) e a emergência dos recintos de fossos no SW peninsular nos finais do 4º milénio A.N.E.

Tese de doutoramento, Arqueologia, Faculdade de Ciências Humanas e Sociais, Universidade do Algarve, 2015