Siderophore Production by Bacillus megaterium: Effect of Growth Phase and Cultural Conditions

Siderophore production by Bacillus megaterium was detected, in an iron-deficient culture medium, during the exponential growth phase, prior to the sporulation, in the presence of glucose; these results suggested that the onset of siderophore production did not require glucose depletion and was not related with the sporulation. The siderophore production by B. megaterium was affected by the carbon source used. The growth on glycerol promoted the very high siderophore production (1,182 μmol g−1 dry weight biomass); the opposite effect was observed in the presence of mannose (251 μmol g−1 dry weight biomass). The growth in the presence of fructose, galactose, glucose, lactose, maltose or sucrose, originated similar concentrations of siderophore (546–842 μmol g−1 dry weight biomass). Aeration had a positive effect on the production of siderophore. Incubation of B. megaterium under static conditions delayed and reduced the growth and the production of siderophore, compared with the incubation in stirred conditions.

Polycyclic aromatic hydrocarbons in gas and particulate phases of indoor environments influenced by tobacco smoke: levels, phase distributions, and health risks

As polycyclic aromatic hydrocarbons (PAHs) have a negative impact on human health due to their mutagenic and/or carcinogenic properties, the objective of this work was to study the influence of tobacco smoke on levels and phase distribution of PAHs and to evaluate the associated health risks. The air samples were collected at two homes; 18 PAHs (the 16 PAHs considered by U.S. EPA as priority pollutants, dibenzo[a,l]pyrene and benzo[j]fluoranthene) were determined in gas phase and associated with thoracic (PM10) and respirable (PM2.5) particles. At home influenced by tobacco smoke the total concentrations of 18 PAHs in air ranged from 28.3 to 106 ngm 3 (mean of 66.7 25.4 ngm 3),∑PAHs being 95% higher than at the non-smoking one where the values ranged from 17.9 to 62.0 ngm 3 (mean of 34.5 16.5 ngm 3). On average 74% and 78% of ∑PAHs were present in gas phase at the smoking and non-smoking homes, respectively, demonstrating that adequate assessment of PAHs in air requires evaluation of PAHs in both gas and particulate phases. When influenced by tobacco smoke the health risks values were 3.5e3.6 times higher due to the exposure of PM10. The values of lifetime lung cancer risks were 4.1 10 3 and 1.7 10 3 for the smoking and nonsmoking homes, considerably exceeding the health-based guideline level at both homes also due to the contribution of outdoor traffic emissions. The results showed that evaluation of benzo[a]pyrene alone would probably underestimate the carcinogenic potential of the studied PAH mixtures; in total ten carcinogenic PAHs represented 36% and 32% of the gaseous ∑PAHs and in particulate phase they accounted for 75% and 71% of ∑PAHs at the smoking and non-smoking homes, respectively.

Construção de um variador de frequência

A dificuldade de controlo de um motor de indução, bem como o armazenamento de energia CC e posterior utilização como energia alternada promoveram o desenvolvimento de variadores de frequência e inversores. Assim, como projeto de tese de mestrado em Automação e Sistemas surge o desenvolvimento de um variador de frequência. Para elaboração do variador de frequência efetuou-se um estudo sobre as técnicas de modulação utilizadas nos inversores. A técnica escolhida e utilizada é a Sinusoidal Pulse Width Modulation (SPWM). Esta técnica baseia-se na modelação por largura de impulso (PWM), o qual é formado por comparação de um sinal de referência com um sinal de portadora de elevada frequência. Por sua vez, a topologia escolhida para o inversor corresponde a um Voltage Source Inverter (VSI) de ponte trifásica completa a três terminais. O desenvolvimento da técnica de modulação SPWM levou ao desenvolvimento de um modelo de simulação em SIMULINK, o qual permitiu retirar conclusões sobre os resultados obtidos. Na fase de implementação, foram desenvolvidas placas para o funcionamento do variador de frequência. Assim, numa fase inicial foi desenvolvida a placa de controlo, a qual contém a unidade de processamento e que é responsável pela atuação de Insulated Gate Bipolar Transistors (IGBTs). Para além disso, foi desenvolvida uma placa para proteção dos IGBTs (evitando condução simultânea no mesmo terminal) e uma placa de fontes isoladas para alimentação dos circuitos e para atuação dos IGBTs. Ainda, foi desenvolvida a técnica de SPWM em software para a unidade de controlo e finalmente foi desenvolvida uma interface gráfica para interação com o utilizador. A validação do projeto foi conseguida através da variação da velocidade do motor de indução trifásico. Para isso, este foi colocado a funcionar a diversas frequências de funcionamento e a diferentes amplitudes. Para além disso, o seu funcionamento foi também validado utilizando uma carga trifásica equilibrada de 3 lâmpadas de forma a ser visualizada a variação de frequência e variação de amplitude.

Influência da actividade do tronco inferior na sequência de activação muscular proximal durante o passo (fase de apoio) - em crianças com quadro motor diplegia

A marcha assegura uma progressão do corpo, compatível com o equilíbrio dinâmico e adaptada a potenciais factores destabilizadores, de um ponto de vista antecipatório, através de sinergias coordenadas entre os MSs, o tronco e os MIs. O tronco inferior tem um papel preponderante na marcha, sobretudo na estabilização necessária durante a fase de apoio. Esta actividade implica mobilidade pélvica e alongamento activo dos abdominais para conseguir a relação comprimento-tensão muscular óptima entre quadricípite e isquiotibiais, permitindo uma correcta sequência, timing e amplitude de activação. Nas crianças com alterações neuromotoras existem alterações no controlo do movimento e na estrutura do próprio movimento, alterando todo este processo. Como tal, este estudo tem como principal objectivo determinar a influência da actividade do tronco inferior na activação muscular proximal durante a fase de apoio da marcha, em crianças com quadro motor de diplegia, caracterizada por uma dificuldade na relação entre os membros e entre estes e o tronco. Para responder a este objectivo realizou-se um estudo de série de casos, com 2 crianças com quadro motor de diplegia. Efectuou-se EMG dos músculos abdominais, quadricípite e isquiotibiais e análise de imagem (para amplitude da CF) durante a marcha, em ambos os membros e em dois momentos de avaliação, separados por 2 meses, nos quais se realizou um protocolo de intervenção terapêutica adequado a cada caso. Os resultados indicam que a variação de amplitude da CF desde a fase de ataque ao solo à fase média de apoio é aproximadamente igual em M0e M1; concretamente, a amplitude inicial é inferior à de referência (pouca flexão) (melhor em M0) e a amplitude final é superior à de referência (pouca extensão) (melhor em M1). Estes resultados são idênticos em ambos os casos. Na EMG verificou-se uma actividade mais global e sincronizada de todos os músculos, mantendo-se aproximadamente a mesma percentagem de activação em M1, sobretudo no caso 1. No caso 2 verificou-se uma maior eficiência na variação da percentagem de activação dos abdominais, em M1, e dos isquiotibiais, à direita. Em conclusão, pode dizer-se que, em crianças com alterações neuromotoras (quadro motor de diplegia), uma actividade mais eficiente e sincronizada no tempo do tronco inferior, nomeadamente dos abdominais, contribui para uma maior capacidade de extensão da CF, durante a fase de apoio.

Comfort and functionality of pregnant women's feet study of kinetic parameters with silicon insoles

As gestantes, fruto das suas alterações fisiológicas e biomecânicas, constituem uma população de risco relativamente a dores ou lesões do sistema músculo-esquelético, nomeadamente, nos membros inferiores e coluna. Os objectivos deste estudo consistiram em avaliar: (i) a dor e o conforto dos pés durante a marcha: sem o uso de qualquer palmilha nas gestantes e no grupo de controlo; com a aplicação de uma palmilha de retropé e com a aplicação de uma palmilha completa (nas gestantes); (ii) a distribuição das pressões plantares e, (iii) as forças de reacção do solo nas mesmas condições experimentais. Avaliámos ainda a duração das diferentes fases do ciclo de marcha nas gestantes, com e sem palmilhas, e no grupo de controlo, sem o uso de palmilha. Os nossos resultados mostraram que: (i) as gestantes demoram mais tempo a completar a fase de apoio da marcha, (ii) têm um aumento significativo de dores nos pés, face ao grupo de controlo, (iii) as gestantes sentem menos dor e mais conforto quando realizam marcha, com palmilhas, especialmente com a palmilha completa, (iv) a palmilha completa redistribui as forças, diminui os valores de pressão e aumenta a área de contacto do pé com o solo. Os nossos resultados sugerem que, o uso da palmilha completa de silicone, durante a marcha, pode ser eficaz na melhoria da sintomatologia dolorosa e no aumento do conforto da grávida.

Thermodynamic pathway for the formation of SnSe and SnSe2 polycrystalline thin films by selenization of metal precursors

In this work, tin selenide thin films (SnSex) were grown on soda lime glass substrates by selenization of dc magnetron sputtered Sn metallic precursors. Selenization was performed at maximum temperatures in the range 300 °C to 570 °C. The thickness and the composition of the films were analysed using step profilometry and energy dispersive spectroscopy, respectively. The films were structurally and optically investigated by X-ray diffraction, Raman spectroscopy and optical transmittance and reflectance measurements. X-Ray diffraction patterns suggest that for temperatures between 300 °C and 470 °C, the films are composed of the hexagonal-SnSe2 phase. By increasing the temperature, the films selenized at maximum temperatures of 530 °C and 570 °C show orthorhombic-SnSe as the dominant phase with a preferential crystal orientation along the (400) crystallographic plane. Raman scattering analysis allowed the assignment of peaks at 119 cm−1 and 185 cm−1 to the hexagonal-SnSe2 phase and those at 108 cm−1, 130 cm−1 and 150 cm−1 to the orthorhombic-SnSe phase. All samples presented traces of condensed amorphous Se with a characteristic Raman peak located at 255 cm−1. From optical measurements, the estimated band gap energies for hexagonal-SnSe2 were close to 0.9 eV and 1.7 eV for indirect forbidden and direct transitions, respectively. The samples with the dominant orthorhombic-SnSe phase presented estimated band gap energies of 0.95 eV and 1.15 eV for indirect allowed and direct allowed transitions, respectively.

Differences in Mismatch Negativity (MMN) response to Pure-tone and Speech sounds in normal subjects: an additional explanation

The relation of automatic auditory discrimination, measured with MMN, with the type of stimuli has not been well established in the literature, despite its importance as an electrophysiological measure of central sound representation. In this study, MMN response was elicited by pure-tone and speech binaurally passive auditory oddball paradigm in a group of 8 normal young adult subjects at the same intensity level (75 dB SPL). The frequency difference in pure-tone oddball was 100 Hz (standard = 1 000 Hz; deviant = 1 100 Hz; same duration = 100 ms), in speech oddball (standard /ba/; deviant /pa/; same duration = 175 ms) the Portuguese phonemes are both plosive bi-labial in order to maintain a narrow frequency band. Differences were found across electrode location between speech and pure-tone stimuli. Larger MMN amplitude, duration and higher latency to speech were verified compared to pure-tone in Cz and Fz as well as significance differences in latency and amplitude between mastoids. Results suggest that speech may be processed differently than non-speech; also it may occur in a later stage due to overlapping processes since more neural resources are required to speech processing.

PAH air pollution at a Portuguese urban area: carcinogenic risks and sources identification

This study aimed to characterize air pollution and the associated carcinogenic risks of polycyclic aromatic hydrocarbon (PAHs) at an urban site, to identify possible emission sources of PAHs using several statistical methodologies, and to analyze the influence of other air pollutants and meteorological variables on PAH concentrations.The air quality and meteorological data were collected in Oporto, the second largest city of Portugal. Eighteen PAHs (the 16 PAHs considered by United States Environment Protection Agency (USEPA) as priority pollutants, dibenzo[a,l]pyrene, and benzo[j]fluoranthene) were collected daily for 24 h in air (gas phase and in particles) during 40 consecutive days in November and December 2008 by constant low-flow samplers and using polytetrafluoroethylene (PTFE) membrane filters for particulate (PM10 and PM2.5 bound) PAHs and pre-cleaned polyurethane foam plugs for gaseous compounds. The other monitored air pollutants were SO2, PM10, NO2, CO, and O3; the meteorological variables were temperature, relative humidity, wind speed, total precipitation, and solar radiation. Benzo[a]pyrene reached a mean concentration of 2.02 ngm−3, surpassing the EU annual limit value. The target carcinogenic risks were equal than the health-based guideline level set by USEPA (10−6) at the studied site, with the cancer risks of eight PAHs reaching senior levels of 9.98×10−7 in PM10 and 1.06×10−6 in air. The applied statistical methods, correlation matrix, cluster analysis, and principal component analysis, were in agreement in the grouping of the PAHs. The groups were formed according to their chemical structure (number of rings), phase distribution, and emission sources. PAH diagnostic ratios were also calculated to evaluate the main emission sources. Diesel vehicular emissions were the major source of PAHs at the studied site. Besides that source, emissions from residential heating and oil refinery were identified to contribute to PAH levels at the respective area. Additionally, principal component regression indicated that SO2, NO2, PM10, CO, and solar radiation had positive correlation with PAHs concentrations, while O3, temperature, relative humidity, and wind speed were negatively correlated.

Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells

Bone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.

Bridging the gap between closed and open data. System proposal for the Portuguese Legislation

Esta dissertação apresenta uma proposta de sistema capaz de preencher a lacuna entre documentos legislativos em formato PDF e documentos legislativos em formato aberto. O objetivo principal é mapear o conhecimento presente nesses documentos de maneira a representar essa coleção como informação interligada. O sistema é composto por vários componentes responsáveis pela execução de três fases propostas: extração de dados, organização de conhecimento, acesso à informação. A primeira fase propõe uma abordagem à extração de estrutura, texto e entidades de documentos PDF de maneira a obter a informação desejada, de acordo com a parametrização do utilizador. Esta abordagem usa dois métodos de extração diferentes, de acordo com as duas fases de processamento de documentos – análise de documento e compreensão de documento. O critério utilizado para agrupar objetos de texto é a fonte usada nos objetos de texto de acordo com a sua definição no código de fonte (Content Stream) do PDF. A abordagem está dividida em três partes: análise de documento, compreensão de documento e conjunção. A primeira parte da abordagem trata da extração de segmentos de texto, adotando uma abordagem geométrica. O resultado é uma lista de linhas do texto do documento; a segunda parte trata de agrupar os objetos de texto de acordo com o critério estipulado, produzindo um documento XML com o resultado dessa extração; a terceira e última fase junta os resultados das duas fases anteriores e aplica regras estruturais e lógicas no sentido de obter o documento XML final. A segunda fase propõe uma ontologia no domínio legal capaz de organizar a informação extraída pelo processo de extração da primeira fase. Também é responsável pelo processo de indexação do texto dos documentos. A ontologia proposta apresenta três características: pequena, interoperável e partilhável. A primeira característica está relacionada com o facto da ontologia não estar focada na descrição pormenorizada dos conceitos presentes, propondo uma descrição mais abstrata das entidades presentes; a segunda característica é incorporada devido à necessidade de interoperabilidade com outras ontologias do domínio legal, mas também com as ontologias padrão que são utilizadas geralmente; a terceira característica é definida no sentido de permitir que o conhecimento traduzido, segundo a ontologia proposta, seja independente de vários fatores, tais como o país, a língua ou a jurisdição. A terceira fase corresponde a uma resposta à questão do acesso e reutilização do conhecimento por utilizadores externos ao sistema através do desenvolvimento dum Web Service. Este componente permite o acesso à informação através da disponibilização de um grupo de recursos disponíveis a atores externos que desejem aceder à informação. O Web Service desenvolvido utiliza a arquitetura REST. Uma aplicação móvel Android também foi desenvolvida de maneira a providenciar visualizações dos pedidos de informação. O resultado final é então o desenvolvimento de um sistema capaz de transformar coleções de documentos em formato PDF para coleções em formato aberto de maneira a permitir o acesso e reutilização por outros utilizadores. Este sistema responde diretamente às questões da comunidade de dados abertos e de Governos, que possuem muitas coleções deste tipo, para as quais não existe a capacidade de raciocinar sobre a informação contida, e transformá-la em dados que os cidadãos e os profissionais possam visualizar e utilizar.

Practical software improve design and performance of a randomly packed air stripping column for removing volatile organic compounds from water

STRIPPING is a software application developed for the automatic design of a randomly packing column where the transfer of volatile organic compounds (VOCs) from water to air can be performed and to simulate it’s behaviour in a steady-state. This software completely purges any need of experimental work for the selection of diameter of the column, and allows a choice, a priori, of the most convenient hydraulic regime for this type of operation. It also allows the operator to choose the model used for the calculation of some parameters, namely between the Eckert/Robbins model and the Billet model for estimating the pressure drop of the gaseous phase, and between the Billet and Onda/Djebbar’s models for the mass transfer. Illustrations of the graphical interface offered are presented.

S100A6 Amyloid Fibril Formation Is Calcium-modulated and Enhances Superoxide Dismutase-1 (SOD1) Aggregation

S100A6 is a small EF-hand calcium- and zinc-binding protein involved in the regulation of cell proliferation and cytoskeletal dynamics. It is overexpressed in neurodegenerative disorders and a proposed marker for Amyotrophic Lateral Sclerosis (ALS). Following recent reports of amyloid formation by S100 proteins, we investigated the aggregation properties of S100A6. Computational analysis using aggregation predictors Waltz and Zyggregator revealed increased propensity within S100A6 helices HI and HIV. Subsequent analysis of Thioflavin-T binding kinetics under acidic conditions elicited a very fast process with no lag phase and extensive formation of aggregates and stacked fibrils as observed by electron microscopy. Ca2+ exerted an inhibitory effect on the aggregation kinetics, which could be reverted upon chelation. An FT-IR investigation of the early conformational changes occurring under these conditions showed that Ca2+ promotes anti-parallel β-sheet conformations that repress fibrillation. At pH 7, Ca2+ rendered the fibril formation kinetics slower: time-resolved imaging showed that fibril formation is highly suppressed, with aggregates forming instead. In the absence of metals an extensive network of fibrils is formed. S100A6 oligomers, but not fibrils, were found to be cytotoxic, decreasing cell viability by up to 40%. This effect was not observed when the aggregates were formed in the presence of Ca2+. Interestingly, native S1006 seeds SOD1 aggregation, shortening its nucleation process. This suggests a cross-talk between these two proteins involved in ALS. Overall, these results put forward novel roles for S100 proteins, whose metal-modulated aggregation propensity may be a key aspect in their physiology and function.

The transport of carboxylic acids and important role of the Jen1p transporter during the development of yeast colonies

On solid substrates, yeast colonies pass through distinct developmental phases characterized by the changes in pH of their surroundings from acidic to nearly alkaline and vice versa. At the beginning of the alkali phase colonies start to produce ammonia, which functions as a quorum-sensing molecule inducing the reprogramming of cell metabolism. Such reprogramming includes, among others, the activation of several plasma membrane transporters and is connected with colony differentiation. In the present study, we show that colony cells can use two transport mechanisms to import lactic acid: a ‘saturable’ component of the transport, which requires the presence of a functional Jen1p transporter, and a ‘non-saturable’ component (diffusion) that is independent of Jen1p. During colony development, the efficiency of both transport components changes similarly in central and outer colonial cells. Although the lactate uptake capacity of central cells gradually decreases during colony development, the lactate uptake capacity of outer cells peaks during the alkali phase and is also kept relatively high in the second acidic phase. This lactate uptake profile correlates with the localization of the Jen1p transporter to the plasma membrane of colony cells. Both lactic acid uptake mechanisms are diminished in sok2 colonies where JEN1 expression is decreased. The Sok2p transcription factor may therefore be involved in the regulation of non-saturable lactic acid uptake in yeast colonies.