Quantitative image analysis as a tool for Yarrowia lipolytica dimorphic growth evaluation in different culture media

Yarrowia lipolytica, a yeast strain with a huge biotechnological potential, capable to produce metabolites such as γ-decalactone, citric acid, intracellular lipids and enzymes, possesses the ability to change its morphology in response to environmental conditions. In the present study, a quantitative image analysis (QIA) procedure was developed for the identification and quantification of Y. lipolytica W29 and MTLY40-2P strains dimorphic growth, cultivated in batch cultures on hydrophilic (glucose and N-acetylglucosamine (GlcNAc) and hydrophobic (olive oil and castor oil) media. The morphological characterization of yeast cells by QIA techniques revealed that hydrophobic carbon sources, namely castor oil, should be preferred for both strains growth in the yeast single cell morphotype. On the other hand, hydrophilic sugars, namely glucose and GlcNAc caused a dimorphic transition growth towards the hyphae morphotype. Experiments for γ-decalactone production with MTLY40-2P strain in two distinct morphotypes (yeast single cells and hyphae cells) were also performed. The obtained results showed the adequacy of the proposed morphology monitoring tool in relation to each morphotype on the aroma production ability. The present work allowed establishing that QIA techniques can be a valuable tool for the identification of the best culture conditions for industrial processes implementation.

Self-esteem, assertiveness and resilience in adolescents institutionalized

The institutionalization of children and adolescents has been an increasingly visible problem in modern society. Unfavourable socio-economic conditions have been joining the behavior problems and school absenteeism. When the family fails in its competence for education, social security or the Court withdraws the child or adolescent to a host institution. The aim of this research was to characterize self-esteem, assertiveness and resilience of institutionalized adolescents in the northern region of Portugal and to establish associations with these dependent variables and gender, scholar level and duration of the institutionalization. For the purpose of this study a wider questionnaire was carried out, and validated with a smaller group. It was a transversal study following a predominantly quantitative methodology, with a convenience sample. The sample included 101 adolescents (55 female and 46 males) from eight institutions, aged between 11 to 21 years old (average 15.45). For self-esteem the Rosenberg Self-Esteem Scale (Rosenberg, 1965), already validate for Portuguese adolescents, was used. For assertiveness and resilience it was applied the Global Evaluation Scale of Assertiveness and the Global Evaluation Scale of Resilience (Jardim & Pereira, 2006) we previously adapted and validated for adolescents. Collected data was introduced in a SPSS database. A descriptive analysis was done to characterize the sample concerning all the variables. To establish associations between individual factors and dependent variables t test, correlations and non-parametric test were applied. Results indicated a relatively low self-esteem (28.03), with girls having a lower value than boys, without significant differences. No correlations were found between self-esteem and the time in the institution. Assertiveness of the sample is average (23.97) and higher for girls than boys, with a positive significant correlation with the scholar grade. Also the resilience is average (25.97), having girls a little lower mean than boys and no significant differences or correlations were found.

Spatio-temporal modelling of environmental data

Programa Doutoral em Matemática e Aplicações.

O erro médico na imprensa portuguesa: quando os pacientes fazem parte da notícia

Esta investigação aborda a qualidade da produção noticiosa sobre o erro médico, como um fator essencial na construção do conhecimento público sobre o tema, com o objetivo de compreender que características definem tal produção e até que ponto poderão ser explicadas pela periodicidade e orientação editorial dos jornais; que conceito de erro médico é veiculado pela produção noticiosa sobre o tema; e quais são os protagonistas no discurso jornalístico sobre o erro médico. Foram analisadas as edições de três jornais portugueses, de 2008 a 2011, resultando num corpus de 266 (4,2%) artigos, que foram classificados de acordo com as seguintes variáveis: as fontes de informação citadas (o seu estatuto e especialidade, no caso dos médicos); os temas que são tratados; as características de enquadramento da informação publicada (tom, género jornalístico; e a presença e número de fontes de informação). Pela análise de conteúdo quantitativa, apurou-se que esse tema está em crescimento, essencialmente com notícias de tom negativo e fontes de informação habitualmente identificadas. Não há evidência para afirmar que a periodicidade e a orientação editorial expliquem as variações dessas características, a não ser relativamente ao número de fontes citadas. Vigoram as notícias centradas nos resultados dos erros (mortes ou lesões), provocados por "erros de omissão" e por "erros de comissão", envolvendo uma diversidade de protagonistas: são, tal como acontece na informação sobre saúde em geral, fontes oficiais e especializadas do campo da saúde. Destacam-se os médicos e os juristas e é dado relevo aos pacientes.

The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.

Formação de professores no programa nacional para a alfabetização na idade certa com uso das TIC: um estudo de caso no ensino fundamental

Dissertação de mestrado em Ciências da Educação (área de especialização em Tecnologia Educativa)