Ecos de la historia peninsular en el teatro barroco. Dos comedias de Tirso de Molina y de António Henriques Gomes

This article presents the comparative analysis of two Iberian plays from the 17th century: Tirso de Molina’s Mari Hernández, la gallega [Mari Hernández, the Galician], written c. 1622-1625 and published in Seville in 1627, and No hay contra el honor poder [There is no power against honour] by António Henriques Gomes, published in Madrid in 1652. This analysis explores the way both plays depict the political power and the image of kings, thus revisiting some aspects of the representation of history in Baroque theatre.Este ensayo propone el análisis comparativo de dos comedias ibéricas del siglo XVII: Mari Hernández, la gallega, de Tirso de Molina (escrita hacia 1622-1625, e impresa en Sevilla en 1627), y No hay contra el honor poder, de António Henriques Gomes (publicada en Madrid en 1652). El modo en que se retrata en ambas comedias el poder político y la figura del rey constituye la perspectiva central de la lectura, lo que permitirá revisitar aspectos de la representación de la historia en el teatro barroco.

The role of coagulase-negative Staphylococci biofilms on late-onset sepsis: current challenges and emerging diagnostics and therapies

Infections are one of the most significant complications of neonates, especially those born preterm, with sepsis as one of the principal causes of mortality. Coagulase-negative staphylococci (CoNS), a group of staphylococcal species that naturally inhabit healthy human skin and mucosa, are the most common cause of late-onset sepsis, especially in preterms. One of the risk factors for the development of CoNS infections is the presence of implanted biomedical devices, which are frequently used for medications and/or nutrient delivery, as they serve as a scaffold for biofilm formation. The major concerns related to CoNS infections have to do with the increasing resistance to multiple antibiotics observed among this bacterial group and biofilm cells’ increased tolerance to antibiotics. As such, the treatment of CoNS biofilm-associated infections with antibiotics is increasingly challenging and considering that antibiotics remain the primary form of treatment, this issue will likely persist in upcoming years. For that reason, the development of innovative and efficient therapeutic measures is of utmost importance. This narrative review assesses the current challenges and emerging diagnostic tools and therapies for the treatment of CoNS biofilm-associated infections, with a special focus on late-onset sepsis.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the project with the reference EXPL/BIA-MIC/0032/2021.info:eu-repo/semantics/publishedVersio

Affinity chromatography in plasmid DNA purification for therapeutic applications

The discovery of disease-related genes and the possibility to manipulate the gene set-up in some organisms has fostered the development of innovative human DNA therapeutics over the last years. Although viral vectors are used in the majority of the trials, non-viral vectors, particularly plasmid DNA (pDNA) vectors, are attracting considerable attention as biotherapeutics both in gene therapy or DNA vaccination, due to their lower immunogenicity, toxicity and also the economic, safer and easier manufacture. Nevertheless, it is well known that the success of gene transfer to cells and subsequent expression is strictly affected by the pDNA manufacturing process. The use of pDNA-based therapeutics relies on procedures that efficiently purify the most biologically active and effective topology, the supercoiled (sc) plasmid conformation. However, chromatographic purification of sc pDNA has specific problems which are mostly related to the structural nature of this biomolecule, the resemblance between pDNA topologies and also with some host impurities, as well as the lack of capacity and selectivity of the traditional bead adsorbents. Recently, sc pDNA purification strategies that use amino acids as immobilized ligands have yielded interesting results. Thus, the present work intends to explore and understand the underlying interactions responsible for the biorecognition of sc conformation by the amino acids ligands already used for pDNA purification as well as to establish the elution conditions that favor the prevalence of some interactions on other. By performing some fundamental studies with oligonucleotides it was observed the involvement of several elementary interactions with the amino acids matrices studied, such as hydrophobic, ring-stacking, cation-π, water-mediated bonds, multiple hydrogen bonds, van der Waals and electrostatic interactions. Although hydrophobic interactions easily appear with histidine matrix or ionic interactions with arginine matrix, it was verified the presence of other interactions in function of the elution conditions used. These results were useful for the implementation of a new affinity chromatographic strategy with the amino acid lysine for efficiently and selectively purify the sc pDNA isoform. Lysineaffinity matrix allowed the elimination of the E. coli impurities as well as other ineffective plasmid isoforms present in a complex clarified lysate meeting all the regulatory requirements. The preferential retention of the nucleic acids with higher bases exposure indicates that this matrix can be more suitable for RNA purification. In accordance with the traditional supports limitations, the alternative monolithic chromatography revealed satisfactory affinity properties with excellent mass transfer and capacity characteritics, allowing a fast and efficient plasmid isoforms separation without flow rate dependence. The similar elution conditions with histidine-agarose and the presence of canbonyldiimidazole groups suggested that the imidazole ring present in this monolithic disk is the major responsible in the specific biorecognition of sc pDNA isoform. Integration of monolithic chromatography in the pDNA manufacturing process also increase the global yield of pDNA xviiirecovery for 89%, with a purity degree according to the recommendations of the regulatory agencies that was reflected in the high transfection efficiency of sc pDNA sample on eukaryotic cells (59% in COS-7 cells). Overall, this doctoral research work revealed that amino acid-based affinity chromatography is a powerful and versatile methodology for nucleic acids purification, mainly the sc pDNA topology, guaranteeing suitable purity degree for DNA-based therapies. Besides the selectivity and specificity obtained with amino acids affinity ligands, the application of the innovative monolithic technology in the pDNA purification field brought a great improvement of the speed, resolution and capacity to the chromatographic performance, being a promising association for plasmid purification technology. Hence, this work provided valuable and helpful information concerning amino acid-affinity chromatography and chromatographic supports that can be useful in the future pDNA bioseparation either for preparative and analytical processes.Work financed by the Portuguese Foundation for Science and Technology (SFRH/BD/41390/2007) under the programe QREN - POPH - Type 4.1 – Advanced Training, cofunded by the European Social Fund and by national funds from the MCTES

Analysis of gene expression variability in Staphylococcus epidermidis biofilms

Staphylococcus epidermidis, a normal inhabitant of a human skin and mucosa, has emerged as one of the principle bacterial agents involved in nosocomial infections, particularly, in patients with indwelling medical devices. It’s pathogenesis is related with the ability to adhere and form biofilms on the surface of those medical devices and is also associated with patients’ immune system that can be compromised. This pathologic condition leads to a high morbidity and, uncommonly, mortality. In the last decade, the quantification of gene expression has been one of the major areas of research in progress. The use of molecular biology techniques, such as quantitative (q) PCR, allowed the study of the process of biofilm formation, which is very complex and involves elaborated genetic regulation. When determining the quantification of genetic expression, there are two critical experimental steps that can impact the outcome of the experiment: RNA extraction, traditionally considered the crucial step of the whole experience, and reverse transcriptase reaction. We have previously shown that in S. epidermidis biofilms, RNA extraction procedure has a strong influence on the outcome of gene expression quantification. Here we evaluated the individual contributions of all experimental steps in the outcome of reliable gene expression determinations. To achieve that, we determined the expression of aap, fmtC and lrgB genes using the type strain RP62A, by performing technical duplicates of each experimental step, and evaluating thecoefficient of variability. Interestingly, our results showed that the bulk of the variability of the gene expression quantification derived from the biological replicates, and not from any of the experimental steps. Furthermore, variability from RNA extraction was not significantly different from the variability obtained from reverse transcriptase or qPCR experiments. This study further confirms that biofilms are difficult biological samples with enhanced difficulties in gene expression determinations

Symptoms of temporomandibular joint disorders in university students : a correlational study

Poster presented at the 3rd International Congress of CiiEM - Research and Innovation in Human and Health Sciences. 20-22 June 2018, Campus Egas Moniz, Monte de Caparica, PortugalN/

Can commercial horse or sheep blood replace fresh human blood in an ex vivo model to study S. epidermidis virulence?

[Excerpt] Staphylococcus epidermidis, a commensal bacterium of healthy human skin and mucosae, can cause serious bloodstream infections such as bacteremia and sepsis. These infections are very hard to cure with current antimicrobial strategies and, thus, it is urgent to find new treatment options. To do so, the study of S. epidermidis virulence factors is of utmost importance. Therefore, the ex vivo human blood model has gained special interest because it enables the study of S. epidermidis behavior in the context of a bloodstream infection. However, this model presents limitations, mainly related to the availability of donors, complicating its implementation in the academic context. To overcome this limitation, the possibility of replacing fresh human blood by commercial blood from other mammals was evaluated. [...]This study was supported by FCT through the funded project PTDC/BIA-MOL/29553/2017, under the scope of COMPETE2020 (POCI-01-0145-FEDER-029553) and by the strategic funding of unit UID/BIO/04469/2019.info:eu-repo/semantics/publishedVersio

Transcriptome mining to identify molecular markers for the diagnosis of Staphylococcus epidermidis bloodstream infections

Bloodstream infections caused by Staphylococcus epidermidis are often misdiagnosed since no diagnostic marker found so far can unequivocally discriminate true infection from sample contamination. While attempts have been made to find genomic and/or phenotypic differences between invasive and commensal isolates, possible changes in the transcriptome of these isolates under in vivo-mimicking conditions have not been investigated. Herein, we characterized the transcriptome, by RNA sequencing, of three clinical and three commensal isolates after 2 h of exposure to whole human blood. Bioinformatics analysis was used to rank the genes with the highest potential to distinguish invasive from commensal isolates and among the ten genes identified as candidates, the gene SERP2441 showed the highest potential. A collection of 56 clinical and commensal isolates was then used to validate, by quantitative PCR, the discriminative power of the selected genes. A significant variation was observed among isolates, and the discriminative power of the selected genes was lost, undermining their potential use as markers. Nevertheless, future studies should include an RNA sequencing characterization of a larger collection of isolates, as well as a wider range of conditions to increase the chances of finding further candidate markers for the diagnosis of bloodstream infections caused by S. epidermidis.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of the unit [UIDB/04469/2020]. S.B. was supported by a fellowship granted by a Doctoral Advanced Training [NORTE-69-2015-15] funded by the European Social Fund under the scope of NORTE2020.info:eu-repo/semantics/publishedVersio

O ensino de Português como Língua Adicional na WEB : uma análise multimodal

Dissertação (mestrado)—Universidade de Brasília, Instituto de Letras, Departamento de Linguística, Português e Línguas Clássicas, Programa de Pós-Graduação em Linguística, 2017.Esta dissertação tem como objetivo analisar o site ‘loecsen.com’ como possível potencial no ensino de Português como Língua Adicional (PLA) por meio da análise de clusters (Baldry & Thibault, 2006) e da ressemiotização (Iedema, 2003). Propor-se-ão reflexões a respeito das características linguísticas existentes no loecsen.com, numa perspectiva discurso-crítica, sobre a importância do ensino mediado no ambiente virtual, considerando que ele dispõe de diversos subsídios que circulam na íntegra como forma de instrumentos para auxiliar os estudantes no desenvolvimento de competências e subcompetências comunicativas e de aprendizagem dos conteúdos de PLA. Os corpora presentes na dissertação foram constituídos de dados extraídos do próprio site loecsen.com. A pesquisa ocupou-se das direções qualitativas de Ferraz (2015), Flick (2009) e Chizzoti (2003). A análise apresentou sua base conforme as seguintes assertivas: 1. Quais recursos semióticos são empregados em websites de ensino de PLA?; 2. O que muda de uma página impressa para uma webpage em termos de língua? e, 3. Qual o potencial significativo de um website no ensino de PLA? A fundamentação teórica teve contribuição da Teoria da Multimodalidade e da Teoria Semiótica (Barton & Lee, 2015), (Kress, 1997), (Iedema, 2003), (Barton, 2015), (Ferraz, 2015), que aliadas deram toda a base teórica que sustentou esta pesquisa em sua íntegra. As concepções de página impressa e webpage apresentadas amadureceram nossas percepções sobre realidade virtual e também sobre a relevância da pesquisa voltada exclusivamente para o ensino mediado pela internet. As semioses exploradas pelo site foram, mais precisamente, as seguintes: oral, escrita e visual, e as utilizadas conforme as normas da Teoria da Multimodalidade, quando usadas uma para complementar a outra gerando por meio dessa interação o sentido almejado. As semioses foram aplicadas ao contexto de aula do site, mas ele não soube aproveitá-las, favorecendo apenas uma aprendizagem periférica e mecânica. A análise do website loecsen.com como mediador de ensino/aprendizagem de português como língua adicional não se revelou um suporte relevante dentro da exploração cada vez mais assídua do ciberespaço pela inexistente interação semiótica e modal que são essenciais na construção de sentido.This dissertation aims to analyze the site 'loecsen.com' as a potential in the teaching of Portuguese as an Additional Language (PLA) through the analysis of clusters (Baldry & Thibault, 2006) and resemiotization (Iedema, 2003). It will be proposed reflections on the linguistic characteristics of loecsen.com, in a discourse-critical perspective, on the importance of mediated education in the virtual environment, which has several subsidies that circulate in full as a form of instruments for students in the development of communicative skills and subcompetences and learning PLA contents. The corpora of the dissertation were constituted of data extracted from the own site loecsen.com. The research focused on the qualitative directions of Ferraz (2015), Flick (2009) and Chizzoti (2003). The analysis presented its basis according to the following assertions: 1. What semiotic resources are employed in PLA teaching websites?; 2. What changes from a printed page to a webpage in terms of language? and, 3. What is the significant potential of a website in teaching PLA? The theoretical basis was contributed by the Theory of Multimodality and Semiotic Theory (Barton & Lee, 2015), (Kress, 1997), (Iedema, 2003), (Barton, 2015), (Ferraz, 2015), which allies gave the whole theoretical basis that supported this research in its entirety. The conceptions of printed page and webpage presented matured our conceptions about virtual reality and the relevance of the research focused exclusively on the education mediated by the internet. The semioses were explored by the site, more precisely the semioses: oral, written and visual, and used according to the norms of the Multimodality Theory, how much one used to complement the other generating through this interaction the desired meaning. The semioses were applied to the classroom context of the site, but the site did not know how to take advantage of them, favoring only peripheral and mechanical learning. The analysis of the loecsen.com website, as mediator of teaching / learning of Portuguese language, has not proved to be a relevant support within the exploration of cyberspace, more and more assiduous, by the inexistent semiotic and modal interaction essential in the construction of meaning

Iniciando a construção do processo de aprender a aprender : aprender a escrever textos não literários

Relatório de estágio de mestrado em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino BásicoEste trabalho apresenta o caminho investigativo percorrido durante uma intervenção pedagógica levada a cabo numa turma do 2º ano de escolaridade. A questão que levou à formulação deste projeto de intervenção foi a do interesse em conhecer o impacto de estratégias de intervenção na iniciação dos alunos na aprendizagem dos processos de escrita de texto, tendo como base a competência de aprender a aprender. A intervenção pedagógica desenhada configurou-se como um projeto de investigação-ação e, como tal, teve fins pedagógicos, acima referidos, e fins investigativos. Com efeito, através da realização deste projeto foi meu objetivo compreender, aperfeiçoar e avaliar as minhas práticas pedagógicas na área em estudo, permitindo, deste modo, o desenvolvimento de competências profissionais. A intervenção pedagógica consistiu na implementação de dois ciclos de escrita situados num contexto de comunicação real, durante os quais promovi o ensino e a aprendizagem explícitos da planificação, textualização e revisão de textos. Durante essa intervenção, recorri a diversificados instrumentos de recolha de dados: avaliações iniciais e finais dos conhecimentos dos alunos sobre processos de escrita, a mobilização desses conhecimentos em situações de escrita de texto e momentos de autoavaliação sobre aprendizagens construídas. Uma outra fonte de dados utilizada foi o meu diário reflexivo. A análise dos dados recolhidos, durante este projeto de intervenção, foi predominantemente qualitativa e pontualmente quantitativa. Os resultados obtidos revelaram que as crianças construíram saberes sobre os processos de escrita de texto e conseguiram identificar e nomear essas aprendizagens. Os alunos deram mostras de ter aprendido explicitamente estratégias que poderão mobilizar em futuras situações de escrita. O destinatário e a finalidade dos textos construídos pelas crianças foram os aspetos que trouxeram motivação e significado às aprendizagens. O relato deste projeto termina com uma reflexão sobre o impacto do trabalho por mim realizado no desenvolvimento das minhas competências profissionais, de que destaco o desenvolvimento da própria capacidade reflexiva.This study presents an action research project that was carried out with a second grade class. The intervention project began with the interest of understanding the impact of some pedagogical strategies upon the learning process of writing non-literary texts, taking the competence of learning how to learn as the main cornerstone. Besides these pedagogical aims, the pedagogical intervention had investigative purposes as well, namely to understand, improve and assess the teaching practices in the area under study in order to promote the development of professional competence. The pedagogical intervention consisted on the implementation of two writing cycles situated in a context of real communication, in which the teaching and learning of planning, translating and revision of texts were promoted (Flower & Hayes, 1981). During this intervention, different instruments of data collection were used: initial and final evaluations of students' knowledge about the processes of writing, text composing and self-evaluation about the learning consciously constructed. Another source of data used in the project was the reflective diary. The analysis of data was predominantly qualitative and in some cases quantitative. The results showed that children constructed knowledge about the processes of writing texts and could identify and name such learning. The students revealed having learned explicitly strategies that can be used in future writing situations. The addressee and the purpose of the texts constructed by children brought meaning and motivation to the learning process. The report ends with a reflection about the impact of this practicum project upon the development of teaching professional capacities, with an emphasis upon the development of reflection