Bioinformatic analysis of promoters from Ashbya gossypii

Ashbya gossypii is a filamentous fungus that produces large quantities of riboflavin, a living factory that generates nearly half of this vitamins world supply. The fungus can grow on industrial wastes and can produce other value-added compounds including recombinant proteins, single cell oil, nucleosides, folic acid and organoleptic compounds. At the genome level, 95% of the genes have homologues in Saccharomyces cerevisiae (90% in syntenic positions). Many molecular tools used in yeast are functional in both organisms, including the autonomously replicating sequences. Fine-tuning heterologous metabolic pathways requires the availability of promoters with a wide range of activity. Endogenous promoters provide the main regulatory elements for gene expression control, however, there is a limited range of well characterised promoters available for metabolic engineering of A. gossypii. Repeated copies of promoters for genome editing purposes increases the probability of homologous recombination and causes strain instability, what is disadvantageous when extensive pathway engineering is performed. The identification of transcription factor (TF) binding sites (TFBSs) in promoters is important for rational design of regulatory crosstalk in metabolic engineering strategies. TFs usually bind to degenerated sequences that can be identified using bioinformatics tools. The MEME algorithm can be used to mine DNA motifs from upstream sequences of co-expressed or co-regulated genes. The obtained motifs can further be compared to TFBSs in databases for S. cerevisiae (i.e. JASPAR, TRANSFAC, YEASTRACT). This procedure allowed the identification of 4 DNA motifs, 8 matching putative TFs, and TATA-box as important elements for high level gene expression. Thus, the bioinformatic approach used in this study, in intergenic regions from A. gossypii, can be also performed in sequences of other organisms with the aim of identifying potential candidate motifs for subsequent experimental characterization and allowing future construction of hybrid semi-synthetic promoters.info:eu-repo/semantics/publishedVersio

New genome-editing tools for Ashbya gossypii

Ashbya gossypii is a splendid riboflavin biofactory that suppresses nearly half of the worldwide demand for vitamin B2. The availability of its genome sequence allowed a better understanding of this fungus metabolism and the development of targeted genome editing tools. Such progresses allowed increasing the vitamin B2 natural production levels, and to expand the potential of this fungus towards the production of nucleosides, folic acid, single cell oil and fragrances [1,2]. Now, with a broadening range of potential genetic targets, modular and flexible pathway assembly tools are more convenient than ever. There is also an urgent need for a larger set of well characterized promoters for metabolic flux optimization towards improved production of target products. As such, this work aimed at developing a modular, flexible and fast pathway assembly platform for targeted chromosomal integration in A. gossypii that offers a wide range of expression strengths (promoters). The developed pathway assembly platform is in an early-stage, designed to easily allow fast cloning and exchange of selection markers, promoters, genes and terminators. As endogenous promoters provide the required regulatory elements for gene expression, several transcriptomic datasets were scanned for medium to high strength promoters. A set of uni- and bi-directional promoters was selected, and each promoter cloned into promoter-probes for characterization in terms of expression strength and regulation, a task that is under way using multiple reporter systems. Preliminary results revealed that the bi-directional promoter AgCCW12/HOG1p drives moderate gene expression through the HOG1p side and strong expression through the CCW12p side. The datasets were also used for transcription factor binding motif mining, in order to find important regulation elements that confer strong gene expression values. Five transcription factor binding motifs were identified as strong targets for promoter engineering. This information can hereafter enable the rational design of synthetic/semi-synthetic minimal promoters. With the current genetic engineering turnaround time to genetically modify A. gossypii being a limiting factor to establish this fungus as a state-of-the-art microbial factory, this pathway assembly platform has great relevance. With a growing number of promoters characterized, it will become increasingly easier to genetically channel the A. gossypii metabolism towards the production of value-added chemicals.Study supported by FCT, Compete2020 and Portugal2020 through the strategic funding of UIDB/04469/2020 and Project EcoBioInks4SmartTextiles (PTDC/CTM-TEX/30298/2017–POCI-01-0145-FEDER-030298)info:eu-repo/semantics/publishedVersio

O perfil do Turista que visita o concelho de Faro: relatório de estágio no Município de Faro - serviço de marketing e promoção turística

Atualmente, o turismo tem uma enorme importância e constitui um dos principais fatores de desenvolvimento de localidades, regiões e países. Para que haja um correto desenvolvimento é importante que haja um planeamento rigoroso. Para a sustentabilidade do destino turístico e em função dos turistas e das suas preferências. Neste contexto o estudo do perfil do turista que visita um destino é crucial para o seu desenvolvimento. Este relatório descreve as atividades de estágio realizado no Município de Faro, na Divisão de Desenvolvimento Económico – Serviço de Marketing e Promoção Turística onde, entre outras tarefas desenvolvidas, se procedeu à aplicação de dois questionários, elaborados pela Câmara Municipal de Faro, com o objetivo de analisar o perfil do turista que visita Faro. Um deles aplicado aos turistas alojados em estabelecimentos turísticos localizados em Faro e o outro, realizado anualmente pelo Município aplicado no centro histórico de Faro e nos locais mais turísticos na época de Natal e passagem de ano. Procede-se à análise crítica destes questionários e informação recolhida com forma de definição do perfil do turista que procura a cidade de Faro. O relatório enfatiza ainda a colaboração na elaboração de um guia truístico online de Faro, o Odyssea e a importância da formação realizada sobre “O Potencial Turístico de Faro”.Currently, tourism has great importance and is one of the main factors of development of localities, regions and countries. For proper development, it’s important that there exists strict planning to assure the sustainability of the touristic destination and meet the preferences of the tourists who visit it. In this context the study of the profile of the tourist that visits a destination is crucial for its development. This report describes the activities carried out on my internship in the Municipality of Faro, in the Division of Economic Development - Marketing and Tourism Promotion Service, where, among other tasks, I developed two surveys in order to analyse and settle the profile of the tourist that visits Faro: a) one of them applied to the tourists staying in touristic establishments located in Faro; b) and the other, carried out annually by the Municipality, applied at the historical centre of Faro and in the most tourist places during the periods of Christmas and New Year’s. Following, the critical analysis of these questionnaires was made and the information collected allowed defining the profile of the tourists that choose the city of Faro to visit. The present report also highlights my participation on the development of an online touristic guide to Faro city, the Odyssea and as well, the importance of the training I took “The Tourist Potencial of Faro”

Mixirica show: proposta de anti-programa para uma emissora inexistente

The Mixirica Show is an anti-program designed for a non-existent broadcaster that proposes a universe centered on dialogues between music and pop culture, using the most varied experiments and references ranging from affective memory to contemporary times, using surrealism and of dadaism.O Mixirica Show é um anti-programa elaborado para uma emissora inexistente que propõe um universo centrado em diálogos entre a música e a cultura pop, utilizando as mais variadas experimentações e referências que vão desde a memória afetiva até a contemporaneidade passando pelo surrealismo e o dadaísmo

Enhanced bacterial cellulose production in komagataeibacter sucrofermentans: impact of different pqq-dependent dehydrogenase knockouts and ethanol supplementation

Background Bacterial cellulose (BC) is a biocompatible material with unique mechanical properties, thus holding a significant industrial potential. Despite many acetic acid bacteria (AAB) being BC overproducers, cost-effective production remains a challenge. The role of pyrroloquinoline quinone (PQQ)-dependent membrane dehydrogenases (mDH) is crucial in the metabolism of AAB since it links substrate incomplete oxidation in the periplasm to energy generation. Specifically, glucose oxidation to gluconic acid substantially lowers environmental pH and hinders BC production. Conversely, ethanol supplementation is known to enhance BC yields in Komagataeibacter spp. by promoting efficient glucose utilization. Results K. sucrofermentans ATCC 700178 was engineered, knocking out the four PQQ-mDHs, to assess their impact on BC production. The strain KS003, lacking PQQ-dependent glucose dehydrogenase (PQQ-GDH), did not produce gluconic acid and exhibited a 5.77-fold increase in BC production with glucose as the sole carbon source, and a 2.26-fold increase under optimal ethanol supplementation conditions. In contrast, the strain KS004, deficient in the PQQ-dependent alcohol dehydrogenase (PQQ-ADH), showed no significant change in BC yield in the single carbon source experiment but showed a restrained benefit from ethanol supplementation. Conclusions The results underscore the critical influence of PQQ-GDH and PQQ-ADH and clarify the effect of ethanol supplementation on BC production in K. sucrofermentans ATCC 700178. This study provides a foundation for further metabolic pathway optimization, emphasizing the importance of diauxic ethanol metabolism for high BC production.Fundação para a Ciência e Tecnologiainfo:eu-repo/semantics/publishedVersio

A Influência da certificação de qualidade na performance das grandes empresas portuguesas

O evoluir dos encerramentos de empresas com o repetir das crises financeiras faz com que seja cada vez mais importante o estudo dos efeitos das decisões na vida das empresas. A incessante busca pela qualidade, bem com a certificação da mesma, via International Standard Organization (ISO), do ponto de vista teórico, apresenta uma possível relação com o pressuposto da continuidade, possibilitando o potenciar da sustentabilidade económico-financeira da empresa, algo que procuraremos avaliar.Assim, a fim de continuar a disseminação das técnicas, possibilitar uma visão global dos modelos e complementar estudos já desenvolvidos, aprofundando a investigação sobre a falência e performance, com o potencial de poder vir a melhorar os modelos, selecionámos, as empresas portuguesas com dimensão grande (segundo a recomendação 2003/361/CE), com Código de Actividade Económica (CAE) D e G, certificadas com ISO 9000 ou 9001, totalizando 91. Foram ainda geradas as Empresas Médias de cada um destes sectores.Às amostras foram aplicadas técnicas uni e multivariadas (MDA), modelos de previsão de falência empresarial, observados em estudos anteriores como os mais eficientes para a economia ibérica, segundo Peres e Antão (2018) e Peres e Antão (2019), a fim de aferir a performance das empresas, bem como a esperada supremacia das que detêm certificação de qualidade.Como principal conclusão, quer para as técnicas de análise uni como na multivariada, não se observa uma performance superior das empresas certificadas em relação às demais, pelo que não se identifica a efectiva implicação da certificação de qualidade na optimização do desempenho economico-financeiro das empresas

Ghrelin expression in human and rat fetal lungs and the effect of ghrelin administration in nitrofen-induced congenital diaphragmatic hernia

Ghrelin is a strong physiologic growth hormone secretagogue that exhibits endocrine and non-endocrine actions. In this study, ghrelin expression in humans and rats was evaluated throughout development of normal and hypoplastic lungs associated with congenital diaphragmatic hernia (CDH). Additionally, the effect of antenatal treatment with ghrelin in the nitrofen-induced CDH rat model was tested. In normal lungs, ghrelin was expressed in the primitive epithelium at early stages of development and decreased in levels of expression with gestational age. In hypoplastic lungs ghrelin was overexpressed in both human and rat CDH fetuses when compared with controls. Exogenous administration of ghrelin to nitrofen-treated dams led to an attenuation of pulmonary hypoplasia of CDH pups. Furthermore, the growth hormone, secretagogue receptor (GHSR1a), could not be amplified from human or rat fetal lungs by RT-PCR. In conclusion, of all the lungs studied so far, the fetal lung is one of the first to express ghrelin during development and might be considered a new source of circulating fetal ghrelin. Overexpression of ghrelin in hypoplastic lungs and the effect of exogenous administration of ghrelin to nitrofen-treated dams strongly suggest a role for ghrelin in mechanisms involved in attenuation of fetal lung hypoplasia, most likely through a GHSR1a-independent pathway

Temas de investigação em direitos humanos para o século XXI

Edição comemorativa do 10.º aniversário do Mestrado em Direitos Humanos da Universidade do Minho.Este livro é uma celebração do ensino e da investigação em direitos humanos que têm vindo a ser desenvolvidos, na Escola de Direito da Universidade do Minho, há já mais de uma década. A sua publicação num momento em que se avolumam os riscos para valores fundamentais subjacentes à proteção dos direitos humanos – como a igualdade e a não discriminação, a proibição da escravatura e de tratamentos cruéis, desumanos e degradantes, a liberdade de religião ou crença, entre muitos outros –, torna-o especialmente oportuno. Os sinais de aparente retrocesso no consenso das nossas sociedades a respeito desses valores – visíveis no triunfo político de discursos abertamente racistas, xenófobos, sexistas, etc. – recordam-nos que, também no mundo ocidental, os direitos humanos são um work in progress, não um dado adquirido. Os novos riscos para a dignidade da pessoa humana associados aos avanços tecnológicos andam de par com velhas formas de subalternização e de opressão. O campo para a reflexão crítica é muito vasto. Os temas que hoje (pre)ocupam académicos, decisores políticos e ativistas de direitos humanos são também aqueles que estruturam o plano de estudos do Mestrado em Direitos Humanos da Universidade do Minho. Todos estes temas surgem ao longo do presente livro, que reúne contributos de muitos dos membros da comunidade científica e académica que o Mestrado em Direitos Humanos mobilizou e ajudou a dinamizar ao longo da última década, entre docentes do Mestrado, colaboradores em júris de provas públicas e/ou na orientação de mestrandos, oradores convidados e estudantes. Os textos aqui reunidos refletem bem as sinergias interdisciplinares, interinstitucionais e inter-nacionais que o Mestrado em Direitos Humanos foi capaz de criar, não apenas pela variedade de campos disciplinares representados – Direito, Filosofia, Relações Internacionais, Antropologia –, mas também pela participação de autores que são docentes e/ou investigadores em diversas instituições nacionais e estrangeiras, como a Faculdade de Direito da Universidade do Porto, a Faculdade de Economia da Universidade de Coimbra, a Universidade Federal da Paraíba e a Pontifícia Universidade Católica de São Paulo.info:eu-repo/semantics/publishedVersio