A Continuidade Pedagógica e a Articulação Curricular entre a Educação Pré-Escolar e o 1º Ciclo do Ensino Básico

Relatório de Estágio apresentado à Escola Superior de Educação de Paula Frassinetti do grau de Mestre em Educação Pré-Escolar e Ensino do 1º Ciclo do Ensino BásicoO relatório é constituído por vários capítulos e centra-se nos conceitos de continuidade educativa e na articulação curricular entre estes dois níveis de ensino. De seguida, é descrita a intervenção nos dois contextos e termina com as considerações finais

Reviewing Social Tourism for Future Research

The purpose of this work is to review the scientific literature (journal articles) that analyses social tourism, with the objective of identifying the evolution of the concept, the methodologies adopted and the main results of the empirical work in this field. The review of literature undertaken provides an analysis of the evolution of published research and also permits the identification of future research pathways

Heterologous production of curcuminoids in E. coli through an artificial biosynthetic pathway

Curcuminoids are natural pigments from plants that have been reported as potential cancer-fighting drugs. Nevertheless, they have a poor bioavailability. Cellular uptake is low, and they are quickly metabolized once inside the cell, requiring repetitive oral doses to achieve sufficient concentration inside the cell for therapeutic activity. The aim of this work is to engineer an artificial biosynthetic pathway for the production of curcuminoids by Escherichia coli. Starting from the substrate tyrosine, the curcumin pathway involves several enzymatic steps: conversion of tyrosine to p-coumaric acid; conversion of p-coumaric acid to caffeic acid; production of caffeoyl-CoA from caffeic acid; production of feruloyl-CoA from caffeoyl-CoA; and finally the production of curcumin from feruloyl-CoA and possibly other curcuminoids, due to enzyme promiscuity. The enzymes involved in the two first enzymatic steps are tyrosine ammonia lyase from Rhodotorula glutinis, P450 CYP199A2 from Rhodopseudomonas palustris, and the redox partners pdr from Pseudomonas putida and pux from R. palustris. Coumaric acid and caffeic acid were successfully produced. A coumaroyl-CoA ligase from Arabidopsis thaliana is being explored for the conversion of the different carboxylic acids into their corresponding CoA esters. Different combinations of this enzyme and caffeoyl-CoA 3-methyl transferase may lead to the production of different curcuminoids. For the last step of the pathway two approaches are being studied: the use of diketide-CoA synthase and curcuminoid synthase from Curcuma longa, and curcumin synthase from Oryza sativa that itself catalyzes both steps. Curcumin and bisdemethoxycurcumin were produced using both approaches and their production was confirmed by HPLC analysis, as well as by the yellow color of the culture supernatant. Successful construction of the curcuminoids biosynthetic pathway would mark a significant step forward in the in situ production of these poorly soluble, anti-carcinogenic compounds

Efeito do grau de superenrolamento de plasmídeos na sua estabilidade estrutural e função biológica

Tese de mestrado em Biologia (Biologia Molecular e Genética), apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2008Actualmente, a utilização de DNA plasmídico (pDNA) como vector em Terapia génica, e em particular nas vacinas de DNA tem vindo a generalizar-se. Estas moléculas podem existir em diferentes conformações com diversos graus de superenrolamento. Várias evidências apontam para uma relação entre este superenrolamento e o sucesso das vacinas de DNA. O objectivo principal deste trabalho foi estudar o efeito do grau de superenrolamento na estabilidade estrutural e função biológica de plasmídeos destinados a terapia génica. Utilizaram-se seis plasmídeos todos derivados do vector comercial pVAX1/LacZ, três dos quais (pVAX1/GFP-BGH, pVAX1/GFP-SV40, pVAX1/GFP-Synt) contêm o gene que codifica a proteína repórter GFP (Green Fluorescent Protein) e os outros três (pVAX1/LacZ-BGH, pVAX1/LacZ-SV40, pVAX1/LacZ-Synt), o gene que codifica a proteína repórter ß-galactosidase. Dentro de cada grupo de três plasmídeos, as diferenças situam-se a nível das sequências de poliadenilação. A utilização de técnicas baseadas na electroforese em géis de agarose das séries completas de topoisómeros, permitiram a determinação do grau de superenrolamento dos plasmídeos utilizados como modelos de estudo. A análise estrutural dos mesmos, por electroforese bidimensional e espectroscopia de dicroísmo circular, não detectou qualquer transição estrutural ou conformação alterada. Estudou-se também a influência do superenrolamento na resistência dos plasmídeos à acção da nuclease S1 e verificou-se que os plasmídeos mais superenrolados são menos resistentes. Por fim, investigou-se o efeito do grau de superenrolamento na função biológica dos vectores plasmídicos e verificou-se que a isoforma superenrolada apresenta sempre percentagens de transfecção superiores à correspondente forma relaxada.The past decade has seen the use of plasmid DNA as a vector for gene therapy, in particular DNA Vaccines, broaden up. These therapeutic molecules may exist in several different conformations, and also in different degrees of supercoiling. Evidences show that there is a relation between the supercoiling and the vaccine's success. The scope of this research was to study the effect of different degrees of supercoiling in the structural stability and biological function of plasmid DNA, to be used in gene therapy. In order to do so, six model plasmids derived from the commercial pVax1/LacZ were used. Three of these plasmids (pVAX1/GFP-BGH, pVAX1/GFP-SV40, pVAX1/GFP-Synt) contained the reporter gene GFP (Green Fluorescent Protein) whereas, the other three (pVAX1/LacZ-BGH, pVAX1/LacZ-SV40, pVAX1/LacZ-Synt), contained the reporter gene ß-galactosidase. Within each group of the referred plasmids the differences lie specifically in the polyadenilation sequences. The use of tecniques based in agarose electrophoresis gels of all the series of topoisomers allowed to determine the degree of supercoiling of each of the plasmids used as a study model. Further structure analysis of the model plasmids by bidimensional electrophoresis and by circular dichroism has not detected any structural transition nor conformation change. The influence of supercoiling in the resistance to nucleases action was also studied, proving that the most supercoiled plasmids are more prone to the action of the S1 nuclease. Finally, the effect of the degree of supercoiling in the biological function of the DNA vaccines was also studied. The latter showed that the supercoiled isoforms of the model plasmids always presented higher transfection percentages than their corresponding relaxed isoform

Design and construction of a new biosynthetic pathway for the production of curcuminoids in Escherichia coli

Curcuminoids are produced by plants and due to their potential as novel cancer-fighting drugs they have recently attracted increased attention. Nevertheless, they have a poor bioavailability. Cellular uptake is low, and they are quickly metabolized once inside the cell, requiring repetitive oral doses to achieve sufficient concentration inside the cell for therapeutic activity. The goal of this PhD project is to engineer a synthetic pathway for curcuminoid in a model bacterium and trigger its release concurrent with ultrasound treatment. The proposed tasks involve several design and engineering steps to program Escherichia coli to execute the new synthetic pathway triggered by a temperature increase. The heat shock response machinery of E. coli will be used as a sensor in the design of the model bacterium. Afterwards, the gene sequences of the enzymes that catalyze each reaction in the curcuminoid pathway will be synthesized and introduced in the E. coli genome applying several cloning strategies. Data from several well documented experiments on E. coli in relevant conditions that have been published were analyzed to select the most expressed heat shock genes in E. coli with the strongest heat shock promoters. The ibpA, dnaK and fxsA gene promoters were chosen based on their induction rates and expression and were validated by RT-qPCR and subsequently through the construction of a stress probe using an adequate reporter gene

Inhibiting and stimulating factors for the integration of refugees into the Portuguese labour market - case studies on best practices within the hospitality sector

Employment is considered a fundamental pillar of the overall refugee integration. The following paper aims at revealing the inhibiting and stimulating factors influencing the integration of refugees into the labour market. It is divided into a collective and an individual part. The collective part starts by providing an overview of the refugee crisis worldwide and in Portugal: then, it zooms in one of the theoretical models of acculturation and concludes by introducing the major factors influencing integration; the individual part investigates seven case studies within the hospitality industry with the intention of providing Mezze with sufficient recommendations and food for thought for its future refugee training program