Evolution of Drug Resistance: Insight on TEM β-Lactamases Structure and Activity and β-Lactam Antibiotics

Since the discovery of the first penicillin bacterial resistance to β-lactam antibiotics has spread and evolved promoting new resistances to pathogens. The most common mechanism of resistance is the production of β-lactamases that have spread thorough nature and evolve to complex phenotypes like CMT type enzymes. New antibiotics have been introduced in clinical practice, and therefore it becomes necessary a concise summary about their molecular targets, specific use and other properties. β-lactamases are still a major medical concern and they have been extensively studied and described in the scientific literature. Several authors agree that Glu166 should be the general base and Ser70 should perform the nucleophilic attack to the carbon of the carbonyl group of the β-lactam ring. Nevertheless there still is controversy on their catalytic mechanism. TEMs evolve at incredible pace presenting more complex phenotypes due to their tolerance to mutations. These mutations lead to an increasing need of novel, stronger and more specific and stable antibiotics. The present review summarizes key structural, molecular and functional aspects of ESBL, IRT and CMT TEM β-lactamases properties and up to date diagrams of the TEM variants with defined phenotype. The activity and structural characteristics of several available TEMs in the NCBI-PDB are presented, as well as the relation of the various mutated residues and their specific properties and some previously proposed catalytic mechanisms.

Modelação e estimação do risco de crédito: estudo de uma carteira

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Matemática e Aplicações - Actuariado, Estatística e Investigação Operacional

Metodologia Seis Sigma: Implementação e Impacto nas PMEs Portuguesas

Dissertação de Mestrado em Gestão Integrada da Qualidade, Ambiente e Segurança

Estimation of infection rate in epidemic models with multiple populations

Dissertação para obtenção do Grau de Mestre em Matemática e Aplicações Especialização em Actuariado, Estatística e Investigação Operacional

Aplicação de variadores de velocidade em sistemas de climatização

Por um lado vivemos numa sociedade cujos padrões de conforto são cada vez mais exigentes, por outro encontramo-nos numa era pautada por um certo declínio económico, social e também ambiental. Não se assumindo uma estratégia que limite ou diminua essas condições de conforto resta-nos atuar de forma a que os recursos utilizados para garantir esse mesmo conforto sejam utilizados da melhor forma possível. O setor dos edifícios é responsável por uma grande parcela de consumo de energia na sociedade atual, sendo que os sistemas de climatização que dele fazem parte, apresentam.se na maioria dos casos como os grandes consumidores de energia. Nesse sentido têm sido feitos esforços enormes no sentido de encontrar soluções que garantam a eficiência energética destes sistemas. O Variador Eletrónico de Velocidade apresenta-se como uma das soluções amplamente difundidas, na vertente do controle de processos e economia de energia. A sua aplicação em sistema de climatização é mais um desses casos. Nesse sentido, numa primeira parte é feito o estudo das características e funcionamento dos Variadores de Velocidade, Sistemas de Climatização e sua aplicação conjunta. Em seguida é realizada uma aplicação informática que pretende demonstrar a economia de energia garantida por aplicação de um Variador de Velocidade.

Biochemistry of grape berries: post-genomics approaches to uncover the effects of water deficits on ripening

Dissertation presented to obtain the Ph.D degree in Biochemistry, Plant Physiology

Classificações geomecânicas mineiras para o dimensionamento de maciços rochosos: o aplicativo MGC-RocDesign|Calc

Este trabalho tem como objetivo destacar a importância da utilização de software de geoengenharia no estudo das classificações de maciços rochosos nas engenharias de minas e geotécnica. Esta investigação pretendeu demonstrar a importância das classificações geomecânicas e índices geológico‐geomecânicos, tais como a Rock Mass Rating (RMR), Rock Tunnelling Quality Index (Q‐system), Surface Rock Classification (SRC), Rock Quality Designation (RQD), Geological Strength Index (GSI) and Hydro‐Potential Value (HP). Para esse efeito foi criada e desenvolvida uma calculadora geomecânica – MGC‐RocDesign|CALC: ‘Mining Geomechanics Classification systems for rock engineering design (version beta)’ – para de certa forma tornar mais simples, rápido e preciso o estudo das classificações geomecânicas sem que seja necessário recorrer manualmente às fastidiosas tabelas das classificações. A MGC‐RocDesign|CALC foi criada e desenvolvida no programa de folha de cálculo Microsoft Excel™ em linguagem Visual Basic for Applications© proporcionando o ambiente de carregamento de dados mais apelativos para o utilizador. Foi ainda integrada neste aplicativo a Calculadora Geotech|CalcTools que resulta da fusão das bases de dados ScanGeoData|BGD e SchmidtData|UCS criadas por Fonseca et al. (2010). Toda a informação foi integrada numa base de dados dinâmica associada a uma plataforma cartográfica em Sistemas de Informação Geográfica. Apresenta‐se como caso de estudo um dos trechos subterrâneos do maciço rochoso da antiga mina de volfrâmio das Aveleiras/Tibães (Mosteiro de Tibães, Braga, NW de Portugal). Além disso, apresenta‐se uma proposta de zonamento geomecânico do maciço rochoso da antiga mina das Aveleiras/Tibães com o objetivo de apoiar o dimensionamento de maciços rochosos. Por fim, apresenta‐se uma reflexão em termos de aplicabilidade, das potencialidades e das limitações da Calculadora Geomecânica MGC‐RocDesign|CALC.

Are societal changes new? Questions or trends and future perceptions on knowledge-based economy

With the emergence of a global division of labour, the internationalisation of markets and cultures, the growing power of supranational organisations and the spread of new information technologies to every field of life, it starts to appear a different kind of society, different from the industrial society, and called by many as ‘the knowledge-based economy’, emphasizing the importance of information and knowledge in many areas of work and organisation of societies. Despite the common trends of evolution, these transformations do not necessarily produce a convergence of national and regional social and economic structures, but a diversity of realities emerging from the relations between economic and political context on one hand and the companies and their strategies on the other. In this sense, which future can we expect to the knowledge economy? How can we measure it and why is it important? This paper will present some results from the European project WORKS – Work organisation and restructuring in the knowledge society (6th Framework Programme), focusing the future visions and possible future trends in different countries, sectors and industries, given empirical evidences of the case studies applied in several European countries, underling the importance of foresight exercises to design policies, prevent uncontrolled risks and anticipate alternatives, leading to different ‘knowledge economies’ and not to the ‘knowled

Molecular typing of dengue virus type 2 in Brazil

Strain typing is a critical tool for molecular epidemiological analysis and can provide important information about the spread of dengue viruses. Here, we performed a molecular characterization of DEN-2 viruses isolated in Brazil during 1990-2000 from geographically and temporally distinct areas in order to investigate the genetic distribution of this serotype circulating in the country. Restriction site-specific polymerase chain reaction (RSS)-PCR presented the same pattern for all 52 Brazilian samples, showing the circulation of just one DEN-2 variant. Phylogenetic analysis using progressive pairwise alignments from 240-nucleotide sequences of the E/NS1 junction in 15 isolates showed that they belong to genotype III (Jamaica genotype).

Analysis of the clonal relationship among clinical isolates of Salmonella enterica serovar Infantis by different typing methods

Salmonella Infantis has been the second most common serovar in Argentina in the last two years, being isolated mostly from paediatric hospitalised patients. In order to determine the clonal relationship among Salmonella Infantis strains, we examined 15 isolates from paediatric patient faeces in Argentina (12 geographically related and 3 geographically non-related) by using antimicrobial susceptibility, plasmid profiling, repetitive extragenic palindromic (REP) PCR, enterobacterial repetitive intergenic consensus (ERIC) PCR, and low-frequency restriction analysis of chromosomal DNA by pulsed field gel electrophoresis (PFGE). Four Spanish strains were included as controls of clonal diversity in molecular techniques. Antibiotype and plasmid profile was not useful as epidemiological tools. PFGE and REP-PCR were able to discriminate between Argentinean and Spanish isolates of Salmonella Infantis allowing to detect genetically related strains in three different cities. This finding indicates that a possible spread of a clone of this serovar in the North-eastern Region of Argentina has taken place in 1998.

Estimation Models for the adoption and use of information technology: internet-related technologies at firm level in the Portuguese and European context

Dissertação apresentada como requisito parcial para obtenção do grau de Doutor em Gestão de Informação

Estimação de Modelos de Adopção e Difusão de Tecnologias de Informação e Comunicação em Portugal

Dissertação apresentada como requisito parcial para obtenção do grau de Mestre em Estatística e Gestão de Informação

New and low cost plastic membrane electrode with low detection limits for sulfadimethoxine determination in aquaculture waters

Sulfadimethoxine (SDM) is one of the drugs, often used in the aquaculture sector to prevent the spread of disease in freshwater fish aquaculture. Its spread through the soil and surface water can contribute to an increase in bacterial resistance. It is therefore important to control this product in the environment. This work proposes a simple and low-cost potentiometric device to monitor the levels of SDM in aquaculture waters, thus avoiding its unnecessary release throughout the environment. The device combines a micropipette tip with a PVC membrane selective to SDM, prepared from an appropriate cocktail, and an inner reference solution. The membrane includes 1% of a porphyrin derivative acting as ionophore and a small amount of a lipophilic cationic additive (corresponding to 0.2% in molar ratio). The composition of the inner solution was optimized with regard to the kind and/or concentration of primary ion, chelating agent and/or a specific interfering charged species, in different concentration ranges. Electrodes constructed with inner reference solutions of 1 × 10−8 mol/L SDM and 1 × 10−4 mol/L chromate ion showed the best analytical features. Near-Nernstian response was obtained with slopes of −54.1 mV/decade, an extraordinary detection limit of 7.5 ng/mL (2.4 × 10−8 mol/L) when compared with other electrodes of the same type. The reproducibility, stability and response time are good and even better than those obtained by liquid contact ISEs. Recovery values of 98.9% were obtained from the analysis of aquaculture water samples.

Solid contact PVC membrane electrodes based on neutral or charged carriers for the selective reading of anionic sulfamethoxazole and their application to the analysis of aquaculture water

Sulfamethoxazole (SMX) is among the antibiotics employed in aquaculture for prophylactic and therapeutic reasons. Environmental and food spread may be prevented by controlling its levels in several stages of fish farming. The present work proposes for this purpose new SMX selective electrodes for the potentiometric determination of this sulphonamide in water. The selective membranes were made of polyvinyl chloride (PVC) with tetraphenylporphyrin manganese (III) chloride or cyclodextrin-based acting as ionophores. 2-nitrophenyl octyl ether was employed as plasticizer and tetraoctylammonium, dimethyldioctadecylammonium bromide or potassium tetrakis (4-chlorophenyl) borate was used as anionic or cationic additive. The best analytical performance was reported for ISEs of tetraphenylporphyrin manganese (III) chloride with 50% mol of potassium tetrakis (4-chlorophenyl) borate compared to ionophore. Nersntian behaviour was observed from 4.0 × 10−5 to 1.0 × 10−2 mol/L (10.0 to 2500 µg/mL), and the limit of detection was 1.2 × 10−5 mol/L (3.0 µg/mL). In general, the electrodes displayed steady potentials in the pH range of 6 to 9. Emf equilibrium was reached before 15 s in all concentration levels. The electrodes revealed good discriminating ability in environmental samples. The analytical application to contaminated waters showed recoveries from 96 to 106%.

"Multiplex PCR" identification of the atypical and monophasic Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- in São Paulo State, Brazil: frequency and antibiotic resistance patterns

Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two) through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.