Study of transcripts and reactive oxygen species involved in the defence responses of Quercus suber against Phytophthora cinnamomi

The present work has the merit of exploring an insight into the activation of defence genes of Quercus suber during response to infection by Phytophthora cinnamomi. Thus, cDNA-AFLP methodology was used to identify gene fragments differentially present in the mRNA profiles of host cells of micropropagated Q. suber plantlets roots infected with zoospores of P. cinnamomi at different post challenge time points. Six candidate genes were selected based on their interesting cDNA-AFLP expression patterns and homology to genes known to play a role in defence. These six genes encode a cinnamyl alcohol dehydrogenase 2 (QsCAD2), a protein disulphide isomerase (QsPDI), a CC-NBS-LRR resistance protein (QsRPc), thaumatin-like protein (QsTLP), chitinase (QsCHI) and a 1,3-beta glucanase (QsGLU). The current work has been successful in evaluation of the expression of these genes by qRT-PCR. Data analysis revealed that transcript levels of QsRPc, QsCHI, QsCAD2 and QsPDI increased during the early hours of inoculation, while transcript profiles of thaumatin-like protein showed decreasing. No expression was detected for 1,3-beta-glucanase (QsGLU). Furthermore, the choice of suitable reference genes in any new experimental system is absolutely crucial in qRT-PCR; for this reason in this study and for the first time a set of potential reference genes were analyzed and validated for qRT-PCR normalization in the patho-system Phytophthora-Q. suber. Four candidate reference genes polimerase II (QsRPII), eukaryotic translation initiation factor 5A(QsEIF-5A), b-tubulin (QsTUB) and a medium subunit family protein of Clathrin adaptor complexes (QsCACs) were evaluated to determine the most stable internal references in Q. suber. Analysis of stability of genes was carried out using Genex software. Results indicated all these four potential reference genes assumed stable expression. Data analysis revealed that QsRPII and QsCACs were the two most stable genes, while genes QsTUB and QsEIF-5A were the third and the fourth most stable gene, respectively. In this study, a plasmid-based quantitative PCR method was developed to measure P. cinnamomi colonization during infection process of Q. suber. Plasmid-based detection of P. cinnamomi showed a gradual accumulation of the pathogen DNA in cork oak root tips up to 24 h post infection. The higher increase in P. cinnamomi/plasmid DNA ratio occurred between 18 and 24 h. One of the primary objectives of this research was to study the effect of cinnamomins (elicitins secreted by P. cinnamomin) on inducing defence mechanism against the pathogen, as recent histological and ultra-structural studies showed that P. cinnamomi was restricted to the outer cortex root fragments pre-treated with capsicien and cryptogein, suggesting that elicitins can stimulate plant defence reactions against P. cinnamomi. To complement these studies and to have a clear view of the nature of the interaction, the role of cinnamomins in the production of the oxidative burst [ROS and ROS scavenging enzymes such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD)] and in the defence responses was evaluated. Cork oak seedlings were pretreated with alpha-cinnamomin and then inoculated with P. cinnamomi mycelia. Results showed a significant higher production of reactive oxygen species (ROS) (H2O2 and O2•-) in elicitin and non-elicitin treated roots in interaction with P. cinnamomi in comparison to the corresponding control. The plant group inoculated with the pathogen after cinnamomin treatment showed an earlier increase in H2O2 production but this was lower as compared with that group inoculated with P. cinnamomi alone. Also, in elicitin pre-treated group generally, a lower level of O2•− production during infection was observed as compared with inoculated roots with P. cinnamomi alone without elicitin treatment. Furthermore, in this study, we evaluated activities of antioxidant enzymes upon challenge with P. cinnamomi, with and without pretreatment with alpha cinnamomin. Results indicated that the activities of defense enzymes POD, SOD and CAT increased after P. cinnamomi inoculation when compared with those in the control group. Also, in the group treated with alpha-cinnamomin followed by P. cinnamomi inoculation, a higher level of enzymatic activities was detected as compared with elicitin non-treated group, which suggest the protective effect of alpha-cinnamomin against the pathogen due to higher elevated levels of defense enzymes POD, SOD and CAT during the infection period. Furthermore, a sensitive qPCR method was applied to measure the pathogen biomass in elicited and non-elicited Q. suber roots challenged with P. cinnamomi to elucidate the effect of cinnamomins on the colonization of P. cinnamomi. Plasmid-based quantification of P. cinnamomi showed a significant decrease in accumulation of the pathogen DNA in cork oak roots after treatment with alpha and beta-cinnamomins which attest the role of cinnamomins in promoting defense responses in cork oak against P. cinnamomi invasion.

Competências parentais percebidas e satisfação marital em famílias com menores em risco

As famílias com menores em risco psicossocial têm que enfrentar várias adversidades que comprometem a sua capacidade para exercer uma parentalidade adequada e frequentemente adotam práticas com efeitos negativos para o desenvolvimento dos seus filhos. As competências parentais percebidas e a capacidade dos cônjuges para comunicar, colaborar e resolver desacordos desempenham um papel crucial para exercer uma parentalidade eficaz. No presente estudo analisamos as relações entre a satisfação marital e as competências parentais percebidas. A amostra é constituída por 112 pais (80 mães e 32 pais), com uma idade média de 36,18 anos, residentes no Algarve. Foram aplicados, através de uma entrevista individual, o Parental Sense of Competence (Johnston & Mash, 1989), a ENRICH Marital Satisfaction Scale (Fowers & Olson, 1993) e um questionário de dados sócio-demográficos e familiares (Nunes, Lemos, Costa, Nunes, & Almeida, 2011). A maioria dos participantes são casados ou vivem em união de facto, integram lares biparentais e estáveis, possuem um baixo nível educativo, estão desempregados e vivem na pobreza. Estas três últimas condições são preocupantes, uma vez que podem comprometer não só o seu bem-estar psicológico, como o desenvolvimento dos seus filhos. Os resultados encontrados sugerem ainda que os pais com uma maior satisfação marital são aqueles que se percebem como mais eficazes e mais satisfeitos com o seu papel de pais. Não observámos diferenças entre pais e mães nas dimensões estudadas. São discutidas as principais necessidades de apoio e de intervenção nestas famílias para serem tomadas em consideração pelos profissionais que trabalham com elas.

Searching biocompounds in marine natural extracts with potential use in tumor prevention and treatment

Cancer is a multistage process characterized by three stages: initiation, promotion and progression; and is one of the major killers worldwide. Oxidative stress acts as initiator in tumorigenesis; chronic inflammation promotes cancer; and apoptosis inactivation is an issue in cancer progression. In this study, it was investigated the antioxidant, antiinflammatory and antitumor properties of hexane, ether, chloroform, methanol and water extracts of five species of halophytes: A. macrostachyum, P. coronopus, J. acutus, C. edulis and A. halimus. Antioxidant activity was assessed by DPPH• and ABTS•+ methods, and the total phenolics content (TPC) was evaluated by the Folin-Ciocalteau method. The anti-inflammatory activity of the extracts was determined by the Griess method, and by evaluating the inhibition of NO production in LPS-stimulated RAW- 264.7 macrophages. The cytotoxic activity of the extracts against HepG2 and THP1 cell lines was estimated by the MTT assay, and the results obtained were further compared with the S17 non-tumor cell line. The induction of apoptosis of J. acutus ether extract was assessed by DAPI staining. The highest antioxidant activities was observed in C. edulis methanol and the J. acutus ether extracts against the DPPH• radical; and J. acutus ether and A. halimus ether extracts against the ABTS•+ radical. The methanol extracts of C. edulis and P. coronopus, and the ether extract of J. acutus revealed a high TPC. Generally the antioxidant activity had no correlation with the TPC. The A. halimus chloroform and P. coronopus hexane extracts demonstrated ability to reduce NO production in macrophages (> 50%), revealing their anti-inflammatory capacity. The ether extract of J. acutus showed high cytotoxicity against HepG2 cancer cells, with reduced cellular viability even at the lowest concentrations. This outcome was significantly lower than the obtained with the non-tumor cells (S17). This result was complemented by the induction of apoptosis.

Estrutura de comunidades de peixes, amostradas por SVSI - sistema de vídeo subaquático iscado, na Costa Sul de Portugal

Desenvolveu-se um Sistema de Vídeo Submerso Iscado (SVSI) viavelmente adaptado para caracterizar comunidades de peixes do substrato rochoso subtidal (forma não-destrutiva e não-invasiva), entre 5 e 50 metros, na costa sul de Portugal (Algarve). Entre Junho e Julho de 2013, amostraram-se 15 pontos na zona Greta (Praia de Faro), divididos em 3 Amostragens (A1, A2, A3) com isco sardinha, mexilhão e sem isco. Captaram-se foto-sequências de intervalo 1 segundo durante 60 minutos e as imagens resultantes processadas qualitativa e quantitativamente para obtenção de resultados. Registaram-se 32 espécies no total. Os tempos de entrada do 1º indivíduo de cada espécie permitiram aferir 30 minutos máximos de filmagem SVSIs para resultados viáveis, enquanto abundâncias por número de indivíduos comprovaram o isco sardinha como mais eficaz e com maior poder atrativo (do que mexilhão). As espécies Coris julis, Diplodus vulgaris, Diplodus cervinus, Diplodus sargus e Serranus cabrilla foram as mais interventivas no estudo, cujo intervalo de tempo entre contagens se adequou em 30 segundos. O estudo de índices de abundâncias e Nmax mostraram não existirem diferenças entre as comunidades ictiológicas de cada tipo de iscagem, contudo existem diferenças dentro da comunidade geral em si do substrato rochoso. A análise alimentar pelo menor tempo de 1ª alimentação indicou Coris julis, Diplodus vulgaris, Serranus cabrilla e Octopus vulgaris como as espécies mais rápidas a morder os iscos. No geral, 69% das espécies preferiram isco sardinha. Comportamentos como a passagem esporádica, territorialidade e interações com a câmara foram as ações dominantes na análise comportamental. Esta técnica e respetiva metodologia foram o primeiro estudo do género nesta região, que de forma simples, se mostrou uma boa ferramenta metodológica para estudos marinhos sobre comunidades, suas preferências alimentares, abundâncias e tempos de reação, aconselhando-se o seu desenvolvimento tecnológico e melhoramento metodológico analítico dos resultados no futuro.

Bio-synthesis of nanosized semiconductors using mine wastes as material sources

This work describes the synthesis of nanosized metal sulfides and respective SiO2 and/or TiO2 composites in high yield via a straightforward process, under ambient conditions (temperature and pressure), by adding to aqueous metals a nutrient solution containing biologically generated sulfide from sulfate-reducing bacteria (SRB). The nanoparticles‘ (NPs) morphological properties were shown not to be markedly altered by the SRB growth media composition neither by the presence of bacterial cells. We further extended the work carried out, using the effluent of a bioremediation system previously established. The process results in the synthesis of added value products obtained from metal rich effluents, such as Acid Mine Drainage (AMD), when associated with the bioremediation process. Precipitation of metals using sulfide allows for the possibility of selective recovery, as different metal sulfides possess different solubilities. We have evaluated the selective precipitation of CuS, ZnS and FeS as nanosized metal sulfides. Again, we have also tested the precipitation of these metal sulfides in the presence of support structures, such as SiO2. Studies were carried out using both artificial and real solutions in a continuous bioremediation system. We found that this method allowed for a highly selective precipitation of copper and a lower selectivity in the precipitation of zinc and iron, though all metals were efficiently removed (>93% removal). This research has also demonstrated the potential of ZnS-TiO2 nanocomposites as catalysts in the photodegradation of organic pollutants using the cationic dye, Safranin-T, as a model contaminant. The influence of the catalyst amount, initial pH and dye concentration were also evaluated. Finally, the efficiency of the precipitates as catalysts in sunlight mediated photodegradation was investigated, using different volumes of dye-contaminated water (150 mL and 10 L). This work demonstrates that all tested composites have the potential to be used as photocatalysts for the degradation of Safranin-T.

Neuroendocrinology of cooperation: the role of neuropeptides on the modulation of mutualistic behaviour of the Indo-Pacific Cleaner Wrasse (Labroides dimidiatus)

Tese de doutoramento, Ciências do Mar, da Terra e do Ambiente, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Development of non-viral vectors for gene therapy for pathologies of the retina

Tese de doutoramento, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2015

Determinação do risco costeiro em arribas, entre as praias de Santa Eulália e Olhos de Água, com recurso a SIG's

Dissertação de mestrado, Engenharia do Ambiente, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Diatom-based characterization of iberian coastal environments at different time scales

Tese de doutoramento, Geologia (Geologia Económica e do Ambiente), Universidade de Lisboa, Faculdade de Ciências, 2014

Development of new methodologies for C-H insertion

Tese de doutoramento, Farmácia (Química Farmacêutica e Terapêutica), Universidade de Lisboa, Faculdade de Farmácia, 2014

Nanotecnologia aplicada a formulações tópicas de fármacos anti-inflamatórios não esteróides

Tese de doutoramento, Farmácia (Tecnologia Farmacêutica), Universidade de Lisboa, Faculdade de Farmácia, 2014

New tools for chemical modification of proteins

Tese de doutoramento, Farmácia (Química Farmacêutica e Terapêutica), Universidade de Lisboa, Faculdade de Farmácia, 2014

Controlo cognitivo na memória episódica : dados de adolescentes e jovens adultos

Dissertação de mestrado, Ciência Cognitiva, Universidade de Lisboa, Faculdade de Psicologia, Faculdade de Medicina, Faculdade de Ciências, Faculdade de Letras, 2014

O espaço português : a consolidação de um reino cristão ibérico : "a reconquista cristã em Portugal" : lendas e mitos, uma proposta didática"

Relatório da Prática de Ensino Supervisionada, Mestrado em Ensino de História e Geografia no 3º ciclo do Ensino Básico e Secundário, Universidade de Lisboa, 2014

Avaliação do potencial clínico de novos ligandos para o receptor estrogénico em diagnóstico e terapia de tumores da mama

Tese de doutoramento, Química (Química Inorgânica), Universidade de Lisboa, Faculdade de Ciências, 2015