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.

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.

Relatório de estágio: mestrado em ensino da música

Resumo I - O projecto “Piano e outros Instrumentos/orquestra” consiste na implementação do formato Piano e Orquestra em complemento às aulas individuais e aos ensaios de conjunto. O trabalho foi dirigido a um grupo de três alunos com idades de 9, 12 e 16 anos, do curso de Iniciação, Básico e Secundário da EMNSC (Escola de Música de Nossa Senhora do Cabo). Foram criados momentos distintos em cada fase. Estimulou-se o desenvolvimento de competências musicais e de estratégias metacognitivas visando a evolução dos alunos e a eficácia na realização do estudo individual. O Projecto foi criado com o objectivo de desenvolver as competências musicais, a motivação para realização do estudo individual e de grupo, o espírito e a audição crítica, o controlo dos factores psicológicos, a ansiedade, o aumento da autoconfiança, a aprendizagem, a melhoria da atitude positiva no momento da execução pública e o incremento do interesse pelas actividades relacionadas com a prática musical. A realização deste projecto visou analisar o grau de motivação, o desenvolvimento e a aquisição de competências musicais dos alunos de piano que frequentaram semanalmente aulas individuais, aulas extra de preparação, ensaios com Orquestra “Da Capo”, ensaios com a Orquestra Maior, duos com violoncelo e com canto. Um dos objectivos era comparar o grau de motivação e desenvolvimento dos alunos quando estudam obras nas aulas individuais que constituem o programa curricular e, por outro lado, quando o fazem em peças seleccionadas especificamente para este Projecto. Esta experiência visou proporcionar um conhecimento nos diversos domínios da música, uma visão diferente da funcionalidade do piano em conjunto com os outros instrumentos, a descoberta das sonoridades do piano contrastante e ao mesmo tempo, integrante com outros instrumentos, as formas de funcionamento dos instrumentos de corda, de sopro, dos assuntos relacionados com a afinação e pretendendo-se assim apurar o sentido de fazer parte integrante de um todo.

An integrated measure of student perceptions of feedback, engagement and school identification

Teacher feedback influences student learning, identity construction and trajectories. This study tests the measurement properties of a questionnaire designed to assess (a) student perceptions about teacher feedback; (b) student identification with school, and; (c) student engagement. 1089 students in grades 6 through 10 (mean age 13.4) participated in the study. Factor analyses yield dimensions of School Identification, Effective Feedback, Person-Centered Feedback, Engagement, and Social Acceptance. Internal consistency for principal dimensions varied between .77 and .89. The instrument is suitable for assessing student school identification, behavioral engagement, and perceptions of teacher feedback.

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.

Modalidades síncronas e assíncronas no ensino da radiologia do sistema nervoso: estilos e resultados de aprendizagem

Lidar com as problemáticas da saúde, implica um domínio dos processos cognitivos (raciocínio, resolução de problemas e tomada de decisão) e de desempenhos práticos, o que obriga a afectação de um conjunto de atitudes e comportamentos específicos. Este estudo, implementou e avaliou o impacto de experiências pedagógicas desenvolvidas com os estudantes da unidade curricular Radiologia do Sistema Nervoso (RSN) da Escola Superior de Tecnologia da Saúde de Lisboa (ESTeSL). Aplicaram-se metodologias de ensino mistas (presenciais e virtuais) utilizadas na leccionação teórica e prática no ano curricular 2008/2009. Para a avaliação do perfil de aprendizagem dos estudantes foi aplicado o método de Honey & Munford e para a avaliação e monitorização dos conhecimentos aplicaram-se check list baseadas nos conteúdos programáticos. A monitorização das ferramentas da plataforma moodle complementaram a restante informação. Verificou-se uma progressão de aprendizagem positiva para um grupo de estudantes maioritariamente do estilo reflexivo (média=10,6 estudantes). As conclusões apontaram para um impacto positivo quanto à aplicação das metodologias híbridas com maior índice de sucesso para a metodologia assíncrona. Verificou-se também mais flexibilidade no acesso aos conteúdos porém com algumas limitações tais como residência inicial por parte dos estudantes, maior carga de trabalho para os docentes, falta de terminais para acesso à plataforma e pouca experiência de todos os envolvidos no domínio e manipulação da plataforma. ABSTRACT - This study focused on the role of cognitive processes (reasoning, problem solving and decision making) and performance practice in the formation of attitudes and behaviours relating to health issues. It was conducted to evaluate the effects of pedagogical experiences on students who participated in the course in radiography in the Nervous System Imaging Unit (RSN) of the Lisbon Health School of Technology. Mixed (face-to-face and virtual) teaching methodologies were used in theory and practice sessions. Honey and Munford’s method was used to evaluate the learning profile of students. To monitor and evaluate students’ knowledge acquisition, check lists based on program topics were applied. Other information was supplied through the learning platform of Moodle. The student group with mostly a reflective learning style increased their knowledge. The asynchronous method was shown to produce a higher success rate and more flexibility in accessing content but also registered some limitations such as resistance by students, increased workload for teachers, lack of access to the platform and inexperience of all involved in handling the platform.

Comportamentos de interação em alunos com perturbações do espectro do autismo: o contributo de histórias sociais na hora do conto

Dissertação apresentada à Escol a Superior de Educação de Lisboa para obtenção de grau de mestre em Ciências da Educação, especialidade Educação Especial: Problemas Graves de Cognição e Multideficiência

Estudo da percepção dos problemas de usabilidade de um software relacionados com o desempenho dos processos cognitivos

Dissertação apresentada à Escola Superior de Comunicação Social como parte dos requisitos para obtenção de grau de mestre em Audiovisual e Multimédia.

Autoinhibition of TBCB regulates EB1-mediated microtubule dynamics

Tubulin cofactors (TBCs) participate in the folding, dimerization, and dissociation pathways of the tubulin dimer. Among them, TBCB and TBCE are two CAP-Gly domain-containing proteins that together efficiently interact with and dissociate the tubulin dimer. In the study reported here we showed that TBCB localizes at spindle and midzone microtubules during mitosis. Furthermore, the motif DEI/M-COO− present in TBCB, which is similar to the EEY/F-COO− element characteristic of EB proteins, CLIP-170, and α-tubulin, is required for TBCE–TBCB heterodimer formation and thus for tubulin dimer dissociation. This motif is responsible for TBCB autoinhibition, and our analysis suggests that TBCB is a monomer in solution. Mutants of TBCB lacking this motif are derepressed and induce microtubule depolymerization through an interaction with EB1 associated with microtubule tips. TBCB is also able to bind to the chaperonin complex CCT containing α-tubulin, suggesting that it could escort tubulin to facilitate its folding and dimerization, recycling or degradation.

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.

Lighting conditions at an operating room in a Portuguese maternity

Lighting is one of the most important factors in human interaction with the environment. Poor lighting may increase the risk of accidents and could also cause a variety of symptoms including: rapid fatigue, headaches, eyestrain, tired eyes, dry eyes, ocular surface symptoms (watery and irritated eyes), decreased concentration and stress. Specific disorders: degeneration of the sharpness of vision (blurred and double vision) and slowness in changing focus. Apart from the advantages in the health and welfare for the workers, good lighting also leads to better job performance (faster), less errors, better safety, fewer accidents and less absenteeism. The overall effect is: better productivity. Good lighting includes quantity and quality requirements, and should necessarily be appropriate to the activity/task being carried out, bearing in mind the comfort and visual efficiency of the worker.

5' and 3' UTR thymidylate synthase polymorphisms modulate the risk of colorectal cancer independently of the intake of methyl group donors

Thymidylate synthase, as a rate-limiting step in DNA synthesis, catalyses the conversion of dUMP into dTMP using 5,10-methylenotetrahydrofolate as the methyl donor. Two polymorphisms have been described in this gene: a repeat polymorphism in the 5' promoter enhancer region (3R versus 2R) and a 6 bp deletion in the 3' unstranslated region. Both of these may affect protein levels. The present case control study was aimed at investigating the influence of these two polymorphisms on the development of colorectal cancer (CRC), as well as their potential interaction with folate, vitamin B6 and vitamin B12 intake. A total of 196 cases and 200 controls, matched for age and sex distribution, were included in the study. No association was found between CRC and the 28 bp repeat polymorphism, but it was observed that individuals with the 6 bp/del and del/del genotypes had a significantly lower risk of developing the disease (OR=0.47; 95% CI 0.30-0.72). A combined genotype (2R/2R; 6 bp/del+del/del) was also found, which was associated with an even lower risk of developing of the disease (OR=0.42; 95% CI 0.26-0.69). No significant interaction between these polymorphisms and vitamin intake was observed. These results indicate for the first time that the 6 bp/del allele might be a protective factor in the development of CRC, independent of the intake of methyl group donors.