Substrate interaction with recombinant amidase from Pseudomonas aeruginosa during biocatalysis

The interaction of a variety of substrates with Pseudomonas aeruginosa native amidase (E.C. 3.5.1.4), overproduced in an Escherichia coli strain, was investigated using difference FTIR spectroscopy. The amides used as substrates showed an increase in hydrogen bonding upon association in multimers, which was not seen with esters. Evidence for an overall reduction or weakening of hydrogen bonding while amide and ester substrates are interacting with the enzyme is presented. The results describe a spectroscopic approach for analysis of substrate-amidase interaction and in situ monitoring of the hydrolysis and transferase reaction when amides or esters are used as substrates.

Environmental and track perturbations on multiple pantograph interaction with catenaries in high-speed trains

The top velocity of high-speed trains is generally limited by the ability to supply the proper amount of energy through the pantograph-catenary interface. The deterioration of this interaction can lead to the loss of contact, which interrupts the energy supply and originates arcing between the pantograph and the catenary, or to excessive contact forces that promote wear between the contacting elements. Another important issue is assessing on how the front pantograph influences the dynamic performance of the rear one in trainsets with two pantographs. In this work, the track and environmental conditions influence on the pantograph-catenary is addressed, with particular emphasis in the multiple pantograph operations. These studies are performed for high speed trains running at 300 km/h with relation to the separation between pantographs. Such studies contribute to identify the service conditions and the external factors influencing the contact quality on the overhead system. (C) 2013 Elsevier Ltd. All rights reserved.

Effects of copper on the photosynthesis of intact chloroplasts: interaction with manganese

Highly purified, intact chloroplasts were prepared from pea (Pisum sativum L.) and spinach (Spinacia oleracea L.) following an identical procedure, and were used to investigate the cupric cation inhibition on the photosynthetic activity. In both species, copper inhibition showed a similar inhibitor concentration that decreases the enzyme activity by 50% (IC(50) approximately 1.8 microM) and did not depend on the internal or external phosphate (Pi) concentration, indicating that copper did not interact with the Pi translocator. Fluorescence analysis suggested that the presence of copper did not facilitate photoinhibition, because there were no changes in maximal fluorescence (F(m)) nor in basal fluorescence (F(o)) of copper-treated samples. The electron transport through the photosystem II (PSII) was also not affected (operating efficiency of PSII-F'v/F'm similar in all conditions). Yet, under Cu(2+) stress, the proportion of open PSII reaction centers was dramatically decreased, and the first quinone acceptor (Q(A)) reoxidation was fully inhibited, as demonstrated by the constant photochemical quenching (q(P)) along experiment time. The quantum yield of PSII electron transport (Phi(PSII)) was also clearly affected by copper, and therefore reduced the photochemistry efficiency. Manganese, when added simultaneously with copper, delayed the inhibition, as measured by oxygen evolution and chlorophyll fluorescence, but neither reversed the copper effect when added to copper-inhibited plastids, nor prevented the inhibition of the Hill activity of isolated copper-treated thylakoids. Our results suggest that manganese competed with copper to penetrate the chloroplast envelope. This competition seems to be specific because other divalent cations e.g. magnesium and calcium, did not interfere with the copper action in intact chloroplasts. All results do suggest that, under these conditions, the stroma proteins, such as the Calvin-Benson cycle enzymes or others are the most probable first target for the Cu(2+) action, resulting in the total inhibition of chloroplast photosynthesis and in the consequent unbalanced rate of production and consumption of the reducing power.

Concepções e práticas de um centro de actividades ocupacionais

O presente trabalho tem como principal objectivo tentar conhecer e compreender as concepções educativas de um Centro de Actividades Ocupacionais face ao seu público-alvo em particular, as suas dimensões mais relevantes, os factores que as condicionam e os contextos que as limitam ou facilitam. Procurou-se conhecer as concepções e práticas dos actores educativos que intervêm na formação/educação junto de pessoas portadoras de deficiência mental adultas, clientes de um Centro de Actividades Ocupacionais, geograficamente situado em Lisboa. Para tal, elaborou-se um Estudo de Caso utilizando entrevistas semi estruturadas a sete profissionais do centro como técnica de recolha de dados, sendo estes posteriormente tratados através da análise de conteúdo. Assim, definiram-se como objectivos específicos do estudo: caracterizar as finalidades educativas do Centro de Actividades Ocupacionais; caracterizar as práticas de desenvolvimento para os adultos deficientes mentais; caracterizar o modo de participação dos clientes nas actividades: aspectos facilitadores e constrangimentos; identificar as dificuldades dos funcionários do Centro de Actividades Ocupacionais no atendimento aos clientes; caracterizar as iniciativas da instituição face à actualização profissional dos seus funcionários no sentido de fornecer respostas mais adequadas às necessidades dos clientes e, ainda, caracterizar de acordo com o ponto de vista dos actores educativos, a participação das famílias no processo de desenvolvimento dos clientes. Os resultados do estudo evidenciam que as actividades desenvolvidas no Centro de Actividades Ocupacionais se dividem entre as consideradas como socialmente úteis, como a montagem de material eléctrico, por exemplo, e as estritamente ocupacionais. Embora um dos objectivos do Centro de Actividades Ocupacionais seja a inserção do cliente na comunidade, da análise de dados ressalta que não existem muitas actividades que divulguem para o exterior o trabalho desenvolvido pelos adultos portadores de deficiência mental, promovendo pouca interacção dos indivíduos com a comunidade. As profissionais que participaram no estudo revelam motivação para trabalhar junto dos clientes e preocupação em melhorar o seu desempenho profissional, daí sentirem necessidade em adquirir formação contínua e partilhar experiências com outros profissionais. As famílias dos clientes são consideradas pelas participantes do estudo como sendo participativas no quotidiano do seu familiar, contudo existem factores que condicionam essa participação, como o envelhecimento dos progenitores. Abstract The present work aims to know and understand the educational conceptions of an Occupational Activities Center taking into consideration its target audience, particularly, its most relevant dimensions, the conditioning factors and its respective restrictive or facilitating contexts. The objective was to understand the concepts and daily practices of the educational actors that participate in the training/education of the mentally disabled adults who attend an Occupational Activities Center located in Lisbon. In that light, a Case Study was conducted using as the data collection technique semistructured interviews made to seven employees from the Center. This data was later on processed through content analysis. Thus, the following was defined as the study’s specific goals. To: characterize the educational purposes of the Occupational Activities Center; characterize development practices for the mentally disabled adults; characterize the clients’ participation process in the activities and the respective enabling and restrictive aspects; identify the Occupational Activities Center employees’ problems regarding client assistance; characterize the institution’s initiatives regarding the professional update of its employees in order to provide more adequate answers to its clients’ needs and, finally to identify from the educational actors’ perspective the participative role families’ take on when it comes to the clients’ development processes. The study results determine that the activities developed in the Occupational Activities Center are distinguished between the ones considered socially useful, such as the assembly of electric material, and the strictly occupational activities. Although one of the Occupational Activities Center goals is the insertion of the client within the community itself, the data analysis shows that there aren’t many activities that promote to the outside world the work performed by these mentally disabled adults, not advertising enough the individuals’ interaction with the community. The professional women who participated in the study show determination and motivation in working with the clients and also a concern in improving their professional performances, from this derives their need to attend continuous training and to share experiences with other professionals. The study's participants find the clients’ families involved in the daily routines of their relatives. Nonetheless, there are some factors that condition that involvement, such as the parents’ aging.

Studies of the Antiproliferative Activity of Ruthenium (II) Cyclopentadienyl-Derived Complexes with Nitrogen Coordinated Ligands

Four cationic ruthenium(II) complexes with the formula [Ru(eta(5)-C5H5)(PPh3)(2)](+), with L = 5-phenyl-1H-tetrazole (TzH) 1, imidazole (ImH) 2, benzo[1,2-b; 4,3-b'] dithio-phen-2-carbonitrile (Bzt) 3, and [5-(2-thiophen-2-yl)-vinyl]-thiophene-2-carbonitrile] (Tvt) 4 were prepared and characterized in view to evaluate their potentialities as antitumor agents. Studies by Circular Dichroism indicated changes in the secondary structure of ct-DNA. Changes in the tertiary structure of pBR322 plasmid DNA were also observed in gel electrophoresis experiment and the images obtained by atomic force microscopy (AFM) suggest strong interaction with pBR322 plasmid DNA; the observed decreasing of the viscosity with time indicates that the complexes do not intercalate between DNA base pairs. Compounds 1, 2, and 3 showed much higher cytotoxicity than the cisplatin against human leukaemia cancer cells (HL-60 cells).

DNA interaction and cytotoxicity studies of new ruthenium(II) cyclopentadienyl derivative complexescontaining heteroaromatic ligands

Four ruthenium(II) complexes with the formula [Ru(eta(5)-C(5)H(5))(PP)L][CF(3)SO(3)], being (PP = two triphenylphosphine molecules), L = 1-benzylimidazole, 1; (PP = two triphenylphosphine molecules), L = 2,2'bipyridine, 2; (PP = two triphenylphosphine molecules), L = 4-Methylpyridine, 3; (PP = 1,2-bis(diphenylphosphine) ethane), L = 4-Methylpyridine, 4, were prepared, in view to evaluate their potentialities as antitumor agents. The compounds were completely characterized by NMR spectroscopy and their crystal and molecular structures were determined by X-ray diffraction. Electrochemical studies were carried out giving for all the compounds quasi-reversible processes. The images obtained by atomic force microscopy (AFM) suggest interaction with pBR322 plasmid DNA. Measurements of the viscosity of solutions of free DNA and DNA incubated with different concentrations of the compounds confirmed this interaction. The cytotoxicity of compounds 1234 was much higher than that of cisplatin against human leukemia cancer cells (HL-60 cells). IC(50) values for all the compounds are in the range of submicromolar amounts. Apoptotic death percentage was also studied resulting similar than that of cisplatin. (C) 2010 Elsevier Inc. All rights reserved.

Performance of coastal sea-defense infrastructure at El Jadida (Morocco) against tsunami threat: lessons learned from the Japanese 11 March 2011 tsunami

This paper seeks to investigate the effectiveness of sea-defense structures in preventing/reducing the tsunami overtopping as well as evaluating the resulting tsunami impact at El Jadida, Morocco. Different tsunami wave conditions are generated by considering various earthquake scenarios of magnitudes ranging from M-w = 8.0 to M-w = 8.6. These scenarios represent the main active earthquake faults in the SW Iberia margin and are consistent with two past events that generated tsunamis along the Atlantic coast of Morocco. The behavior of incident tsunami waves when interacting with coastal infrastructures is analyzed on the basis of numerical simulations of near-shore tsunami waves' propagation. Tsunami impact at the affected site is assessed through computing inundation and current velocity using a high-resolution digital terrain model that incorporates bathymetric, topographic and coastal structures data. Results, in terms of near-shore tsunami propagation snapshots, waves' interaction with coastal barriers, and spatial distributions of flow depths and speeds, are presented and discussed in light of what was observed during the 2011 Tohoku-oki tsunami. Predicted results show different levels of impact that different tsunami wave conditions could generate in the region. Existing coastal barriers around the El Jadida harbour succeeded in reflecting relatively small waves generated by some scenarios, but failed in preventing the overtopping caused by waves from others. Considering the scenario highly impacting the El Jadida coast, significant inundations are computed at the sandy beach and unprotected areas. The modeled dramatic tsunami impact in the region shows the need for additional tsunami standards not only for sea-defense structures but also for the coastal dwellings and houses to provide potential in-place evacuation.

Particle agglomeration study in rf silane plasmas: In situ study by polarization-sensitive laser light scattering

To determine self-consistently the time evolution of particle size and their number density in situ multi-angle polarization-sensitive laser light scattering was used. Cross-polarization intensities (incident and scattered light intensities with opposite polarization) measured at 135 degrees and ex situ transmission electronic microscopy analysis demonstrate the existence of nonspherical agglomerates during the early phase of agglomeration. Later in the particle time development both techniques reveal spherical particles again. The presence of strong cross-polarization intensities is accompanied by low-frequency instabilities detected on the scattered light intensities and plasma emission. It is found that the particle radius and particle number density during the agglomeration phase can be well described by the Brownian free molecule coagulation model. Application of this neutral particle coagulation model is justified by calculation of the particle charge whereby it is shown that particles of a few tens of nanometer can be considered as neutral under our experimental conditions. The measured particle dispersion can be well described by a Brownian free molecule coagulation model including a log-normal particle size distribution. (C) 1996 American Institute of Physics.