Development of polymeric materials to inhibit bacterial quorum sensing

Bacterial infections are an increasing problem for human health. In fact, an increasing number of infections are caused by bacteria that are resistant to most antibiotics and their combinations. Therefore, the scientific community is currently searching for new solutions to fight bacteria and infectious diseases, without promoting antimicrobial resistance. One of the most promising strategies is the disruption or attenuation of bacterial Quorum Sensing (QS), a refined system that bacteria use to communicate. In a QS event, bacteria produce and release specific small chemicals, signal molecules - autoinducers (AIs) - into the environment. At the same time that bacterial population grows, the concentration of AIs in the bacterial environment increases. When a threshold concentration of AIs is reached, bacterial cells respond to it by altering their gene expression profile. AIs regulate gene expression as a function of cell population density. Phenotypes mediated by QS (QSphenotypes) include virulence factors, toxin production, antibiotic resistance and biofilm formation. In this work, two polymeric materials (linear polymers and molecularly imprinted nanoparticles) were developed and their ability to attenuate QS was evaluated. Both types of polymers should to be able to adsorb bacterial signal molecules, limiting their availability in the extracellular environment, with expected disruption of QS. Linear polymers were composed by one of two monomers (itaconic acid and methacrylic acid), which are known to possess strong interactions with the bacterial signal molecules. Molecularly imprinted polymer nanoparticles (MIP NPs) are particles with recognition capabilities for the analyte of interest. This ability is attained by including the target analyte at the synthesis stage. Vibrio fischeri and Aeromonas hydrophila were used as model species for the study. Both the linear polymers and MIP NPs, tested free in solutions and coated to surfaces, showed ability to disrupt QS by decreasing bioluminescence of V. fischeri and biofilm formation of A. hydrophila. No significant effect on bacterial growth was detected. The cytotoxicity of the two types of polymers to a fibroblast-like cell line (Vero cells) was also tested in order to evaluate their safety. The results showed that both the linear polymers and MIP NPs were not cytotoxic in the testing conditions. In conclusion, the results reported in this thesis, show that the polymers developed are a promising strategy to disrupt QS and reduce bacterial infection and resistance. In addition, due to their low toxicity, solubility and easy integration by surface coating, the polymers have potential for applications in scenarios where bacterial infection is a problem: medicine, pharmaceutical, food industry and in agriculture or aquaculture.

Development of infrared spectroscopy for assessing bacterial quality in foods

Rapid and specific detection of foodborne bacteria that can cause food spoilage or illness associated to its consumption is an increasingly important task in food industry. Bacterial detection, identification, and classification are generally performed using traditional methods based on biochemical or serological tests and the molecular methods based on DNA or RNA fingerprints. However, these methodologies are expensive, time consuming and laborious. Infrared spectroscopy is a reliable, rapid, and economic technique which could be explored as a tool for bacterial analysis in the food industry. In this thesis it was evaluated the potential of IR spectroscopy to study the bacterial quality of foods. In Chapter 2, it was developed a calibration model that successfully allowed to predict the bacterial concentration of naturally contaminated cooked ham samples kept at refrigeration temperature during 8 days. In this part, it was developed the methodology that allowed the best reproducibility of spectra from bacteria colonies with minimal sample preparation, which was used in the subsequent work. Several attempts trying different resolutions and number of scans in the IR were made. A spectral resolution of 4 cm-1, with 32 scans were the settings that allowed the best results. Subsequently, in Chapter 3, it was made an attempt to identify 22 different foodborne bacterial genera/species using IR spectroscopy coupled with multivariate analysis. The principal component analysis, used as an exploratory technique, allowed to form distinct groups, each one corresponding to a different genus, in most of the cases. Then, a hierarchical cluster analysis was performed to further analyse the group formation and the possibility of distinction between species of the same bacterial genus. It was observed that IR spectroscopy not only is suitable to the distinction of the different genera, but also to differentiate species of the same genus, with the simultaneous use of principal component analysis and cluster analysis techniques. The utilization of IR spectroscopy and multivariate statistical analysis were also investigated in Chapter 4, in order to confirm the presence of Listeria monocytogenes and Salmonella spp. isolated from contaminated foods, after growth in selective medium. This would allow to substitute the traditional biochemical and serological methods that are used to confirm these pathogens and that delay the obtainment of the results up to 2 days. The obtained results allowed the distinction of 3 different Listeria species and the distinction of Salmonella spp. from other bacteria that can be mistaken with them. Finally, in chapter 5, high pressure processing, an emerging methodology that permits to produce microbiologically safe foods and extend their shelf-life, was applied to 12 foodborne bacteria to determine their resistance and the effects of pressure in cells. A treatment of 300 MPa, during 15 minutes at room temperature was applied. Gram-negative bacteria were inactivated to undetectable levels and Gram-positive showed different resistances. Bacillus cereus and Staphylococcus aureus decreased only 2 logs and Listeria innocua decreased about 5 logs. IR spectroscopy was performed in bacterial colonies before and after HPP in order to investigate the alterations of the cellular compounds. It was found that high pressure alters bands assigned to some cellular components as proteins, lipids, oligopolysaccharides, phosphate groups from the cell wall and nucleic acids, suggesting disruption of the cell envelopes. In this work, bacterial quantification and classification, as well as assessment of cellular compounds modification with high pressure processing were successfully performed. Taking this into account, it was showed that IR spectroscopy is a very promising technique to analyse bacteria in a simple and inexpensive manner.

Genetic programming and bacterial algorithm for neural networks and fuzzy systems design

In the field of control systems it is common to use techniques based on model adaptation to carry out control for plants for which mathematical analysis may be intricate. Increasing interest in biologically inspired learning algorithms for control techniques such as Artificial Neural Networks and Fuzzy Systems is in progress. In this line, this paper gives a perspective on the quality of results given by two different biologically connected learning algorithms for the design of B-spline neural networks (BNN) and fuzzy systems (FS). One approach used is the Genetic Programming (GP) for BNN design and the other is the Bacterial Evolutionary Algorithm (BEA) applied for fuzzy rule extraction. Also, the facility to incorporate a multi-objective approach to the GP algorithm is outlined, enabling the designer to obtain models more adequate for their intended use.

Genetic and bacterial programming for B-Spline neural networks design

The design phase of B-spline neural networks is a highly computationally complex task. Existent heuristics have been found to be highly dependent on the initial conditions employed. Increasing interest in biologically inspired learning algorithms for control techniques such as Artificial Neural Networks and Fuzzy Systems is in progress. In this paper, the Bacterial Programming approach is presented, which is based on the replication of the microbial evolution phenomenon. This technique produces an efficient topology search, obtaining additionally more consistent solutions.

Design of B-spline neural networks using a bacterial programming approach

The design phase of B-spline neural networks represents a very high computational task. For this purpose, heuristics have been developed, but have been shown to be dependent on the initial conditions employed. In this paper a new technique, Bacterial Programming, is proposed, whose principles are based on the replication of the microbial evolution phenomenon. The performance of this approach is illustrated and compared with existing alternatives.

Alguns romances da tradição oral de Aljezur

As versões de romances que a seguir apresento foram recolhidas no concelho de Aljezur, distrito de Faro, no ano de 1989, quando frequentei a disciplina de Romanceiro Tradicional sob a orientação do Prof. Pere Ferré. Para esta pequena recolha foram preciosos os ensinamentos do Prof. Ferré, e algumas notas sobre a tradição algarvia dadas pela Profª. Vanda Anastácio.

Le traitement de l'erreur à l'oral entre pairs sous l'approche communicative dans une classe de français langue seconde

Ce mémoire aborde le traitement de l'erreur à l'oral entre apprenants dans une classe de français langue seconde de l'Université de Sherbrooke (Québec). Dans les classes de français langue seconde de cette institution, les étudiants travaillent la plupart du temps en équipes du fait que l'approche communicative est utilisée comme modèle pédagogique. L'objectif général de cette recherche a été d'explorer, à partir du moment où une erreur est produite, les relations entre les types de rétroactions utilisées par les apprenants dans le but de corriger leurs collègues et les réponses suivant ces rétroactions. Principalement, cette étude sur le traitement de l'erreur à l'oral dans un contexte de travail en équipe a révélé que la correction faite par un pair ne passe pas inaperçue et qu'elle est fréquemment suivie par une réparation immédiate de l'énoncé fautif initial ou encore, d'une tentative de correction.

Da memória do povo - Recolha de literatura popular da tradição oral do Concelho de Portimão

Margarida Tengarrinha oferece - nos, nesta obra, um leque bastante diversificado do património, adentro das “artes da fala”, correndo nas memórias do povo do concelho de Portimão. O fruto da sua recolecção, os “textos” que recebeu, transcreveu e estudou, é aqui editado com uma valência, cremos,, primeiro que tudo pedagógica. Assim, os textos agrupam - se por “manchas de leitura” com uma coerência interna aglutinadora, aparecendo ordenados sob uma classificação entre formal — segundo os “géneros”, como “contos” , “lendas”, “romances”, “orações”, por exemplo — e temática — por “fundos de sentido” semantizados nas comunicações: “bruxas e bruxedos”, “benzeduras, mezinhas, maldições e superstições”, “poesias maliciosas”, por exemplo.

Sobre a tradição oral Algarvia. 1— Poesia recolhida na freguesia de Querença. As orações

Breve reflexão sobre as orações e os romances-oração contidos na colecção entitulada Tradição Oral Algarvia.I — Poesia Recolhida na Freguesia de Querença. Deixando de lado a perspectiva diacrónica, vamos considerar as formas mnemónicas e formulísticas de algumas orações de protecção: Padre Nosso Pequenino, uma oração da noite, As Tabuínhas de Moisés, uma "oração forte" para ser recitada duma forma extremamente rigorosa e ritualizada, através duma estrita ordem verbal e numérica. É também referido um romance-oração recolhido só no Algarve e que apresenta, em certas versões, formas vestigiais de paralelismo também encontradas nalguns romances líricos.

A estética discursiva nos contos da literatura oral mirandesa: uma abordagem estatístico-pragmática

Neste artigo, pretendo destacar a forma como o discurso dos contos da literatura oral obedece a regras e leis que podem ser medidas e verificadas.Para tal, analisarei três narrativas, em língua mirandesa, recolhidas na Terra de Miranda onde este idioma é falado. Em seguida, procederei à descrição da sua estrutura, assim como à trajectória e organização discursiva, que sobressai para além da aparente simplicidade e desorganização. Na primeira parte, de carácter empírico, descreverei algumas particularidades desta região, reflectindo sobre o papel dos contos da literatura oral na construção da identidade mirandesa. Na segunda, de âmbito estatístico, será feita a análise dos dados lexicais, textuais e discursivos. O método de trabalho baseia-se na estatística paramétrica. Esta análise é confrontada com os dados teóricos que acompanham, corroboram ou contestam, os dados estatísticos. O estudo completa-se com a apresentação dos textos e das ecografias lexicais de cada conto.

La tradición oral en el antiguo testamento. Aproximación al legado de los patriarcas

Há muito que, graças a diversos investigadores (Frazer, Gaster, Patai, etc.), se sabe ter o texto da Bíblia chegado até nós cheio de episódios e motivos que aparecem também nas epopeias de outros povos vizinhos de Israel (o Grande Dilúvio, o menino no cestinho de vime, a Torre de Babel...), todas elas, em geral, anteriores à narrativa bíblica. Mas, deixando de lado a questão das datas e fontes do Pentateuco, assim como da sua relação com as epopeias limítrofes, é possível verificar também que existem episódios que estão, ao mesmo tempo, na Bíblia e na literatura oral moderna e, além disso, que certas passagens bíblicas aproveitaram sequências narrativas que existiam previamente na tradição oral, antes de passarem ao texto escrito. Esta incorporação efectuou-se não de forma inócua, mas sim de acordo com determinada função: validar e justificar origens e actuações. Mostrar este facto é o objectivo do presente artigo (o primeiro duma prevista série de três), em que tratamos sobretudo do episódio de Judá e Tamar (Gén. 38).

Bacterial algorithm applied for fuzzy rule extraction

This paper presents a method of using the so-colled "bacterial algorithm" (4,5) for extracting a fuzzy rule base from a training set. The bewly proposed bacterial evolutionary algorithm (BEA) is shown. In our application one bacterium corresponds to a fuzzy rule system.

Diabetes Mellitus do tipo 2: influência da farmacogenómica na terapêutica oral

Dissertação de mest., Ciências Farmacêuticas, Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

Contributo para o estudo da literatura oral no Algarve

Dissertação de mest., Promoção e Mediação da Leitura, Faculdade de Ciências Humanas e Sociais, Univ. do Algarve, 2012

La qualité du français oral d'étudiants en formation initiale des maîtres regard sur les grilles d'observation

Alors que la didactique du français oral a pris un essor fulgurant au Québec depuis les années 1990, les recherches actuelles n'ont que trop peu effleuré les modalités d'évaluation de la qualité du français oral des futurs enseignants. En effet, ce thème incontournable dans le cadre de la formation des futurs maîtres souffre d'une méconnaissance des diverses modalités d'évaluation de sa qualité par les formateurs universitaires. Le lien entre formation des futurs enseignants et identification de leurs besoins et de leurs difficultés est indéniable. Dans ce contexte, nous avons interrogé les qualités métrologiques de deux grilles d'observation de l'oral : Gervais, Laurier et Paret (1994) ainsi que Préfontaine, Lebrun et Nachbauer (1998). Nous les avons soumises à 10 experts-évaluateurs qui oeuvrent en formation des maîtres et qui ont un lien avec l'évaluation de la qualité du français oral des futurs enseignants.