Bioconversion of D-glucose into D-glucosone by Glucose 2-oxidase from Coriolus versicolor at Moderate Pressures

Glucose 2-oxidase (pyranose oxidase, pyranose: oxygen-2-oxidoreductase, EC 1.1.3.10) from Coriolus versicolor catalyses the oxidation of D-glucose at carbon 2 in the presence of molecular O(2) producing D-glucosone (2-keto-glucose and D-arabino-2-hexosulose) and H(2)O(2). It was used to convert D-glucose into D-glucosone at moderate pressures (i.e. up to 150 bar) with compressed air in a modified commercial batch reactor. Several parameters affecting biocatalysis at moderate pressures were investigated as follows: pressure, [enzyme], [glucose], pH, temperature, nature of fluid and the presence of catalase. Glucose 2-oxidase was purified by immobilized metal affinity chromatography on epoxy-activated Sepharose 6B-IDA-Cu(II) column at pH 6.0. The rate of bioconversion of D-glucose increased with the pressure since an increase in the pressure with compressed air resulted in higher rates of conversion. On the other hand, the presence of catalase increased the rate of reaction which strongly suggests that H(2)O(2) acted as inhibitor for this reaction. The rate of bioconversion of D-glucose by glucose 2-oxidase in the presence of either nitrogen or supercritical CO(2) at 110 bar was very low compared with the use of compressed air at the same pressure. The optimum temperature (55 degrees C) and pH (5.0) of D-glucose bioconversion as well as kinetic parameters for this enzyme were determined under moderate pressure. The activation energy (E(a)) was 32.08 kJmol(-1) and kinetic parameters (V(max), K(m), K(cat) and K(cat)/K(m)) for this bioconversion were 8.8 Umg(-1) protein, 2.95 mM, 30.81 s(-1) and 10,444.06 s(-1)M(-1), respectively. The biomass of C. versicolor as well as the cell-free extract containing glucose 2-oxidase activity were also useful for bioconversion of D-glucose at moderate pressures. The enzyme was apparently stable at moderate pressures since such pressures did not affect significantly the enzyme activity.

Bioconversion of D-glucose into D-glucosone by immobilized glucose 2-oxidase from Coriolus versicolorat moderate pressures

The immobilized glucose 2-oxidase (pyranose oxidase, pyranose:oxygen-2-oxidoreductase, EC 1.1.3.10) from Coriolus versicolor was used to convert D-glucose into D-glucosone at moderate pressures, up to 150 bar, with compressed air in a modified commercial batch reactor. Several parameters affecting biocatalysis at moderate pressures were investigated as follows: pressure, different forms of immobilized biocatalysts, glucose concentration, pH, temperature and the presence of catalase. Glucose 2-oxidase (GOX2) was purified by immobilized metal affinity chromatography on epoxy-activated Sepharose 6B-IDA-Cu(II) column at pH 6.0. Purified enzyme and catalase were immobilized into a polyethersulfone (PES) membrane in the presence of glutaraldehyde and gelatin. Enhancement of the bioconversion of D-glucose was done by the pressure since an increase in the pressure with compressed air increases the conversion rates. The optimum temperature and pH for bioconversion of D-glucose were found to be 62 degrees C and pH 6.0, respectively and the activation energy (E(a)) was 28.01 kJ mol(-1). The apparent kinetic constants (V(max)' K(m)', K(cat)' and K(cat)/K(m)') for this bioconversion were 2.27 U mg(-1) protein, 11.15 mM, 8.33 s(-1) and 747.38 s(-1) M(-1), respectively. The immobilized biomass of C. versicolor as well as crude extract containing GOX2 activity were also useful for bioconversion of D-glucose at 65 bar with a yield of 69.9 +/- 3.8% and 91.3 +/- 1.2%, respectively. The immobilized enzyme was apparently stable for several months without any significant loss of enzyme activity. On the other hand, this immobilized enzyme was also stable at moderate pressures, since such pressures did not affect significantly the enzyme activity. (C) 2010 Elsevier Ltd. All rights reserved.

Evaluating deviations in prostatectomy patients treated with IMRT

Aim - To evaluate the deviations in prostatectomy patients treated with IMRT in order to calculate appropriate margins to create the PTV. Background - Defining inappropriate margins can lead to underdosing in target volumes and also overdosing in healthy tissues, increasing morbidity. Material and methods - 223 CBCT images used for alignment with the CT planning scan based on bony anatomy were analyzed in 12 patients treated with IMRT following prostatectomy. Shifts of CBCT images were recorded in three directions to calculate the required margin to create PTV. Results and discussion - The mean and standard deviation (SD) values in millimetres were −0.05 ± 1.35 in the LR direction, −0.03 ± 0.65 in the SI direction and −0.02 ± 2.05 the AP direction. The systematic error measured in the LR, SI and AP direction were 1.35 mm, 0.65 mm, and 2.05 mm with a random error of 2.07 mm; 1.45 mm and 3.16 mm, resulting in a PTV margin of 4.82 mm; 2.64 mm, and 7.33 mm, respectively. Conclusion - With IGRT we suggest a margin of 5 mm, 3 mm and 8 mm in the LR, SI and AP direction, respectively, to PTV1 and PTV2. Therefore, this study supports an anisotropic margin expansion to the PTV being the largest expansion in the AP direction and lower in SI.

Fusional vergence measurements for eso versus exo deviations: is there a difference?

Purpose: The aims of this study were to compare angle of deviation, fusional vergence measurements and fusion reserve ratio between esophoria and exophoria. Methods: A cross-sectional study was performed in children with best-corrected visual acuity of 0.0 LogMAR in either eye, compensated heterophoria within 10 prism dioptres (PD), full ocular rotations, presence of fusional vergence and stereopsis (60 seconds of arc or better). Fusional amplitudes were compared between angle of deviation (2, 4, 6, 8 and 10 PD) in esophoria and exophoria. The fusion reserve ratio was calculated (to assess the effect of the underlying angle of deviation) as fusional convergence divided by prism alternating cover test measurements. Results: Two-hundred and eleven children (7.65±1.16 years) were recruited to this study. Exophoria was most common for near (n=181; 85.8%) and distance (n=20; 9.5%). Esophoria was present in 22 children for near (10.4%) and in 1 child for distance (0.5%). No significant differences were found between fusional amplitudes and angle of deviation for near (p>0.05). Children with exophoria of 10PD had a slight, but not, significant (p=0.264) increase in fusional convergence from 2PD (19.95±5.09) to 10PD (26.67±5.77). In esophoric children the variation of fusional convergence was smaller from 2P (25.00±0.00) to 10PD (22.50±3.54) and non significant (p=0.185). The fusion reserve ratio was significantly smaller in children with higher deviations (i.e. 10PD) for both esophoria (p=0.003) and exophoria (p>0.001). The fusion reserve ratio ranged between 12.50 (2PD) and 2.25±0.35 (10PD) for esophoria and between 9.98±2.55 (2PD) and 2.67±0.58 (10PD) for exophoria. Conclusions: Angle of deviation is not an efficient measure to predict fusional amplitudes. The fusion reserve ratio appears to be a better measurement to assess the effect of the underlying angle of deviation on fusional convergence. More studies are necessary to understand better the relationship between fusion amplitudes and angle of deviation.

Propriedades psicométricas da versão portuguesa do Kansas City Cardiomyopathy Questionnaire na miocardiopatia dilatada com insuficiência cardíaca congestiva

Vários estudos demonstraram que os doentes com insuficiência cardíaca congestiva (ICC) têm um compromisso da qualidade de vida relacionada com a saúde (QVRS), tendo esta, nos últimos anos, vindo a tornar-se um endpoint primário quando se analisa o impacto do tratamento de situações crónicas como a ICC. Objectivos: Avaliar as propriedades psicométricas da versão portuguesa de um novo instrumento específico para medir a QVRS na ICC em doentes hospitalizados: o Kansas City Cardiomyopathy Questionnaire (KCCQ). População e Métodos: O KCCQ foi aplicado a uma amostra consecutiva de 193 doentes internados por ICC. Destes, 105 repetiram esta avaliação 3 meses após admissão hospitalar, não havendo eventos ocorridos durante este período de tempo. A idade era 64,4± 12,4 anos (entre 21 e 88), com 72,5% a pertencer ao sexo masculino, sendo a ICC de etiologia isquémica em 42%. Resultados: Esta versão do KCCQ foi sujeita a validação estatística semelhante à americana com a avaliação da fidelidade e validade. A fidelidade foi avaliada pela consistência interna dos domínios e dos somatórios, apresentando valores Alpha de Cronbach idênticos nos vários domínios e somatórios ( =0,50 a =0,94). A validade foi analisada pela convergência, pela sensibilidade às diferenças entre grupos e pela sensibilidade à alteração da condição clínica. Avaliou-se a validade convergente de todos os domínios relacionados com funcionalidade, pela relação verificada entre estes e uma medida de funcionalidade, a classificação da New York Heart Association (NYHA), tendo-se verificado correlações significativas (p<0,01), como medida para avaliar a funcionalidade em doentes com ICC. Efectuou-se uma análise de variância entre o domínio limitação física, os somatórios e as classes da NYHA, tendo-se encontrado diferenças estatisticamente significativas (F=23,4; F=36,4; F=37,4; p=0,0001), na capacidade de descriminação da gravidade da condição clínica. Foi realizada uma segunda avaliação em 105 doentes na consulta do 3º mês após a intervenção clínica, tendo-se observado alterações significativas nas médias dos domínios avaliados entre o internamento e a consulta (diferenças de 14,9 a 30,6 numa escala de 0-100), indicando que os domínios avaliados são sensíveis à mudança da condição clínica. A correlação interdimensões da qualidade de vida que compõe este instrumento é moderada, sugerindo dimensões independentes, apoiando a sua estrutura multifactorial e a adequabilidade desta medida para a sua avaliação. Conclusão: O KCCQ é um instrumento válido, sensível à mudança e específico para medir a QVRS numa população portuguesa com miocardiopatia dilatada e ICC. ABSTRACT - Several studies have shown that patients with congestive heart failure (CHF) have a compromised health-related quality of life (HRQL), and this, in recent years, has become a primary endpoint when considering the impact of treatment of chronic conditions such as CHF. Objectives: To evaluate the psychometric properties of the Portuguese version of a new specific instrument to measure HRQL in patients hospitalized for CHF: the Kansas City Cardiomyopathy Questionnaire (KCCQ). Methods: The KCCQ was applied to a sample of 193 consecutive patients hospitalized for CHF. Of these, 105 repeated the assessment 3 months after admission, with no events during this period. Mean age was 64.4±12.4 years (21-88), and 72.5% were 72.5% male. CHF was of ischemic etiology in 42% of cases. Results: This version of the KCCQ was subjected to statistical validation, with assessment of reliability and validity, similar to the American version. Reliability was assessed by the internal consistency of the domains and summary scores, which showed similar values of Cronbach alpha (0.50-0.94). Validity was assessed by convergence, sensitivity to differences between groups and sensitivity to changes in clinical condition. We evaluated the convergent validity of all domains related to functionality, through the relationship between them and a measure of functionality, the New York Heart Association (NYHA) classification. Significant correlations were found (p<0.01) for this measure of functionality in patients with CHF. Analysis of variance between the physical limitation domain, the summary scores and NYHA class was performed and statistically significant differences were found (F=23.4; F=36.4; F=37.4, p=0.0001) in the ability to discriminate severity of clinical condition. A second evaluation was performed on 105 patients at the 3-month follow-up outpatient appointment, and significant changes were observed in the mean scores of the domains assessed between hospital admission and the clinic appointment (differences from 14.9 to 30.6 on a scale of 0-100), indicating that the domains assessed are sensitive to changes in clinical condition. The correlation between dimensions of quality of life in the KCCQ is moderate, suggesting that the dimensions are independent, supporting the multifactorial nature of HRQL and the suitability of this measure for its evaluation. Conclusion: The KCCQ is a valid instrument, sensitive to change and a specific measure of HRQL in a population with dilated cardiomyopathy and CHF.

Asymmetric synthesis of N-aryl aziridines

The reactions of a variety of N-arylhydroxamates as nitrogen transfer reagents to acryloyl derivatives of (−)-8-phenylmenthol, (−)-quinine and (−)-Oppolzer’s sultam acting as Michael acceptors was studied. Poor to modest diastereoselection was observed in the formation of aziridines. The absolute structure of one of the pure diastereomers secured from Oppolzer’s auxiliary was established by X-ray crystallography and hence the absolute configuration of the derived methyl-N-phenylaziridine-2-carboxylate could be assigned. Whilst only poor facial selectivity was observed for chiral hydroxamic acid prepared from dehydroabietic acid, moderate to good enantioselection of aziridines could be achieved with the chiral quaternary salts based on cinchona alkaloids, especially with that of cinchonine. A model is presented to explain the origin of enantioselection and a mechanism is proposed for the aziridination reaction.

Neutron activation analysis of wheat samples

The deficiency of essential micronutrients and excess of toxic metals in cereals, an important food items for human nutrition, can cause public health risk. Therefore, before their consumption and adoption of soil supplementation, concentrations of essential micronutrients and metals in cereals should be monitored. This study collected soil and two varieties of wheat samples–Triticum aestivum L. (Jordão/bread wheat), and Triticum durum L. (Marialva/durum wheat) from Elvas area, Portugal and analyzed concentrations of As, Cr, Co, Fe, K, Na, Rb and Zn using Instrumental Neutron Activation Analysis (INAA) to focus on the risk of adverse public health issues. The low variability and moderate concentrations of metals in soils indicated a lower significant effect of environmental input on metal concentrations in agricultural soils. The Cr and Fe concentrations in soils that ranged from 93–117 and 26,400–31,300 mg/kg, respectively, were relatively high, but Zn concentration was very low (below detection limit <22 mg/kg) indicating that soils should be supplemented with Zn during cultivation. The concentrations of metals in roots and straw of both varieties of wheat decreased in the order of K>Fe>Na>Zn>Cr>Rb>As>Co. Concentrations of As, Co and Cr in root, straw and spike of both varieties were higher than the permissible limits with exception of a few samples. The concentrations of Zn in root, straw and spike were relatively low (4–30 mg/kg) indicating the deficiency of an essential micronutrient Zn in wheat cultivated in Portugal. The elemental transfer from soil to plant decreases with increasing growth of the plant. The concentrations of various metals in different parts of wheat followed the order: Root>Straw>Spike. A few root, straw and spike samples showed enrichment of metals, but the majority of the samples showed no enrichment. Potassium is enriched in all samples of root, straw and spike for both varieties of wheat. Relatively to the seed used for cultivation, Jordão presented higher transfer coefficients than Marialva, in particular for Co, Fe, and Na. The Jordão and Marialva cultivars accumulated not statistically significant different concentrations of different metals. The advantages of using INAA are the multielementality, low detection limits and use of solid samples (no need of digestion).

Manutenção proactiva de sistemas AVAC com recurso aos sistemas inteligentes multiagente

Nos tempos actuais os equipamentos para Aquecimento Ventilação e Ar Condicionado (AVAC) ocupam um lugar de grande importância na concepção, desenvolvimento e manutenção de qualquer edifício por mais pequeno que este seja. Assim, surge a necessidade premente de racionalizar os consumos energéticos optimizando-os. A alta fiabilidade desejada nestes sistemas obriga-nos cada vez mais a descobrir formas de tornar a sua manutenção mais eficiente, pelo que é necessário prevenir de uma forma proactiva todas as falhas que possam prejudicar o bom desempenho destas instalações. Como tal, torna-se necessário detectar estas falhas/anomalias, sendo imprescíndivel que nos antecipemos a estes eventos prevendo o seu acontecimento num horizonte temporal pré-definido, permitindo actuar o mais cedo possível. É neste domínio que a presente dissertação tenta encontrar soluções para que a manutenção destes equipamentos aconteça de uma forma proactiva e o mais eficazmente possível. A ideia estruturante é a de tentar intervir ainda numa fase incipiente do problema, alterando o comportamento dos equipamentos monitorizados, de uma forma automática, com recursos a agentes inteligentes de diagnóstico de falhas. No caso em estudo tenta-se adaptar de forma automática o funcionamento de uma Unidade de Tratamento de Ar (UTA) aos desvios/anomalias detectadas, promovendo a paragem integral do sistema apenas como último recurso. A arquitectura aplicada baseia-se na utilização de técnicas de inteligência artificial, nomeadamente dos sistemas multiagente. O algoritmo utilizado e testado foi construído em Labview®, utilizando um kit de ferramentas de controlo inteligente para Labview®. O sistema proposto é validado através de um simulador com o qual se conseguem reproduzir as condições reais de funcionamento de uma UTA.

Revision of the portuguese catalog of tsunamis

Catastrophic tsunamis are described in historical sources for all regions around the Gulf of Cadiz, at least since 60 BC. Most of the known events are associated with moderate to large earthquakes and among them the better studied is 1 November 1755. We present here a review of the events which effects, on the coasts of the Portuguese mainland and Madeira Island, are well described in historical documents or have been measured by tide gauges since the installation of these instruments. For a few we include new relevant information for the assessment of the tsunami generation or effects, and we discard events that are included in existing compilations but are not supported by quality historical sources or instrumental records. We quote the most relevant quantitative descriptions of tsunami effects on the Portuguese coast, including in all pertinent cases a critical review of the coeval sources, to establish a homogenous event list. When available, instrumental information is presented. We complement all this information with a summary of the conclusions established by paleo-tsunami research.

Volatility regimes for the VIX index

This article presents a Markov chain framework to characterize the behavior of the CBOE Volatility Index (VIX index). Two possible regimes are considered: high volatility and low volatility. The specification accounts for deviations from normality and the existence of persistence in the evolution of the VIX index. Since the time evolution of the VIX index seems to indicate that its conditional variance is not constant over time, I consider two different versions of the model. In the first one, the variance of the index is a function of the volatility regime, whereas the second version includes an autoregressive conditional heteroskedasticity (ARCH) specification for the conditional variance of the index.

A importância do programa preliminar e do projecto no âmbito da contratação pública

O Código dos contratos públicos (CCP) veio introduzir alguns novos conceitos, nas empreitadas de obras públicas, sobre erros e omissões e também sobre trabalhos a mais. Tais conceitos têm estado na origem de algumas polémicas envolvendo donos de obra, projectistas, fiscalizações, empreiteiros e técnicos. Por outro, quer à luz da anterior legislação quer ao abrigo da actual os desvios de preço e prazo, em empreitadas de obras públicas, continuam a ser frequentes, dir-se-á mesmo, praticamente uma constante e frequentemente alvo nos meios de comunicação. No âmbito desta dissertação pretende-se partilhar algumas reflexões emergentes da observação efectuada, como participante activo em diversos empreendimentos. Verificar-se-á ao longo da mesma dissertação que nela é atribuída grande relevância ao programa preliminar constante do Anexo I à Portaria n.º 701-H/2008 de 29 de Julho, que dá origem ao projecto. Com efeito, este documento embora a sua designação de preliminar possa induzir em erro, levando a conferir-lhe pouca importância, é, em boa verdade, o programa fundamental, para um bom entendimento entre o dono de obra e o projectista e subsequentemente com o empreiteiro. Nesta dissertação faz-se uma incursão pelos conteúdos mais relevantes do CCP, no âmbito dos desvios de preço e custo, das empreitadas de obras públicas. Procede-se, também, a uma análise detalhada e sequencial das fases e actividades que contribuem para a concretização de uma obra pública, observando simultaneamente o papel dos diversos intervenientes.

Avaliação da função ventricular esquerda e dimensão da aurícula esquerda por Vscan® e relatório de estágio

Mestrado em Tecnologia de Diagnóstico e Intervenção Cardiovascular. Área de especialização: Ultrassonografia Cardiovascular.

Estudo da função sistólica do ventrículo direito por ecocardiografia transtorácica: relatório de projecto

Mestrado em Tecnologia de Diagnóstico e Intervenção Cardiovascular. Área de especialização - Ultrassonografia Cardiovascular

Deformação miocárdica analisada por Speckle Tracking, em doentes hipertensos com fracção de ejecção preservada

Mestrado em Tecnologia de Diagnóstico e Intervenção Cardiovascular - Área de especialização: Ultrassonografia Cardiovascular.

Relação entre otimismo e autonomia funcional em indivíduos com esclerose múltipla

Objetivo: O presente estudo tem como objetivo verificar a relação existente entre autonomia funcional e otimismo. Método: Os participantes são 280 indivíduos portadores de esclerose múltipla (EM) diagnosticada há mais de um ano em que a maioria são mulheres, com nível elevado de escolaridade, maioritariamente casados, e a trabalhar em regime de tempo integral. A funcionalidade foi avaliada com a Escala de Impacto de Participação e Autonomia – IPA, e o otimismo com o Life Orientation Test – Revised (LOT-R). A recolha de dados realizou-se em unidades de saúde de Lisboa e cumpriu as regras conforme a Declaração de Helsínquia. Conclusão: Os resultados mostram que existem valores de correlação estatisticamente significativos, mas moderados entre as dimensões da LOT-R e IPA, sugerindo que o otimismo é um fator importante para a autonomia funcional dos indivíduos portadores de EM. ABSTRACT - Aim: The present study aims at identifying the relationship between optimism and functional autonomy in individuals with Multiple Sclerosis (MS). Method: The participants are 280 individuals diagnosed with MS for more than one year; the majority are women, with high levels of education, mostly married and employed on full time. To access functionality we used the Impact on Participation and Autonomy Scale – IPA, and to assess optimism the Life Orientation Test - Revised (LOT-R). Data was collected in Lisbon health settings. Data collection follows the rules of the Helsinki Declaration. Conclusion: Results show that correlations between the two main variables are statistically significant, but moderate, suggesting that the perception of optimism is an important factor for functional autonomy in individuals with MS.