Parcerias público-privadas nas políticas sociais. O caso das freguesias da Área Metropolitana de Lisboa (1993-1997)1

PUBLIC-PRIVATE PARTNERSHIPS IN SOCIAL POLICIES. THE CASE OF THEPARISHES IN THE LISBON METROPOLITAN AREA – Capitalist development is currently characterised by an administrative centralisation with the Nation-State losing power to supranational authorities. However, some functions have been decentralised, specially the ones of a social character, to local levels of Public Administration, as can be seen by the increase of parish activities, which develop their own plans, act in partnership or subsidise public and private institutions at a local level. The social policies that the parishes adopt are influenced by several factors, among which we detect the structure of the territory and the dominating political party within the local government council

A importância do plano de trabalhos à luz do novo código dos contratos públicos sob o ponto de vista do empreiteiro

O Plano de Trabalhos é a previsão do desenvolvimento futuro de um empreendimento, devidamente quantificado no tempo, no espaço e nos respectivos custos de execução de uma obra. O Planeamento de uma obra, com a sequência de execução das suas actividades, estabelece os menores dos prazos de execução parciais, a identificação de todas as actividades, referenciando as que são chave em termos de prazos e custos, os meios necessários: financeiros, mão-de-obra, materiais, equipamentos, subempreitadas, etc. A presente dissertação tem como objectivo apresentar uma possível forma de elaborar, controlar, actualizar, alterar, etc., o plano de trabalhos. Apresenta-se seguidamente uma análise do CCP nos artigos que podem ser influenciados pelo plano de trabalhos, nomeadamente os prazos de execução, prorrogações de prazo, multas, suspensões de obra, etc., sob o ponto de vista do Empreiteiro

Interaction between Neisseria gonorrhoeae bacterial peroxidase and its electron donor, the lipid-modified azurin

cofinanced by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728).The Neisseria gonorrhoeae bacterial cytochrome c peroxidase plays a key role in detoxifying the cells from H2O2 by reducing it to water using the lipid-modified azurin, LAz, a small type 1 copper protein, as electron donor. Here, the interaction between these two proteins was characterized by steady-state kinetics, two-dimensional NMR and molecular docking simulations. LAz is an efficient electron donor capable of activating this enzyme. This electron transfer complex is weak with a hydrophobic character, with LAz binding close to the electron transferring heme of the enzyme. The high catalytic rate (39 ± 0.03 s−1) is explained by the LAz pre-orientation, due to a positive dipole moment, and by the fast-dynamic ensemble of orientations, suggested by the small chemical shifts.publishersversionpublishe

YhjA - An Escherichia coli trihemic enzyme with quinol peroxidase activity

Belgian Federal Science Policy Office (Belspo) (grant to BD, IAP7/44, iPROS project). co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728).The trihemic bacterial cytochrome c peroxidase from Escherichia coli, YhjA, is a membrane-anchored protein with a C-terminal domain homologous to the classical bacterial peroxidases and an additional N-terminal (NT) heme binding domain. Recombinant YhjA is a 50 kDa monomer in solution with three c-type hemes covalently bound. Here is reported the first biochemical and spectroscopic characterization of YhjA and of the NT domain demonstrating that NT heme is His63/Met125 coordinated. The reduction potentials of P (active site), NT and E hemes were established to be −170 mV, +133 mV and +210 mV, respectively, at pH 7.5. YhjA has quinol peroxidase activity in vitro with optimum activity at pH 7.0 and millimolar range KM values using hydroquinone and menadiol (a menaquinol analogue) as electron donors (KM = 0.6 ± 0.2 and 1.8 ± 0.5 mM H2O2, respectively), with similar turnover numbers (kcat = 19 ± 2 and 13 ± 2 s−1, respectively). YhjA does not require reductive activation for maximum activity, in opposition to classical bacterial peroxidases, as P heme is always high-spin 6-coordinated with a water-derived molecule as distal axial ligand but shares the need for the presence of calcium ions in the kinetic assays. Formation of a ferryl Fe(IV) = O species was observed upon incubation of fully oxidized YhjA with H2O2. The data reported improve our understanding of the biochemical properties and catalytic mechanism of YhjA, a three-heme peroxidase that uses the quinol pool to defend the cells against hydrogen peroxide during transient exposure to oxygenated environments.publishersversionpublishe

Prenatal Diagnosis of a Congenital Postaxial Longitudinal Limb Defect: A Case Report

Introduction. Although congenital longitudinal fibular deficiency is one of the most common long bone deficiencies, there are few published cases of its prenatal diagnosis. Case report. A right longitudinal deficiency of the fibula associated with tibial shortening, foot equinovalgus, and absence of the fourth and fifth foot rays diagnosed at 22 weeks gestation is described. Sequential ultrasonographic surveillance was performed without obstetric complications. The anomaly was confirmed after birth, and conservative orthopaedic management was decided. Conclusion. Though rarely seen, postaxial longitudinal limb defect may be detected by ultrasound. The correct approach can only be decided after birth, when the functional impact of the anomaly can be fully evaluated

Source and reduction of nitrous oxide

PTDC/BBB-BQB/0129/2014. UID/QUI/50006/2019. UlD/Multi/04378/2019. MSPC acknowledges FCT/MCTES for funding her "Research Position" (signed with FCT NOVA in accordance with DL.57/2016 and Lei 57/2017). Sem PDF conforme despacho.Nitrous oxide is a potent greenhouse gas with a global warming impact 300-fold higher than carbon dioxide. Due to its exponential increase in the atmosphere and its implications in climate change there is the need to develop strategies to mitigate its emissions and to reduce it to the inert dinitrogen gas. Only three enzymes have been reported to be able to reduce nitrous oxide, namely nitrogenase, one multicopper oxidase and nitrous oxide reductase, with the latter being the only one with a relevant physiological activity. In this enzyme, reduction of nitrous oxide occurs in a unique catalytic tetranuclear sulfide center, named “CuZ” center, a complex center required to overcome the high activation barrier of this reaction. Nitrous oxide reductase can be isolated with “CuZ” center in two forms, CuZ*(4Cu1S) and CuZ(4Cu2S), that differ in their catalytic and spectroscopic properties. Recently, another step towards a better understanding of the catalytic and activation mechanism of this enzyme was taken by identifying and spectroscopically characterizing an intermediate species of its catalytic cycle, CuZ 0 . A different approach for N 2 O reduction can be attained using model compounds. The unique structural motif present in “CuZ” center, a Cu 4 (µ 4 -S), has been a challenge for inorganic synthesis but several synthetic clusters that mimic different forms of “CuZ” center have been reported. Model compounds for the oxidation states involved in N 2 O reduction are also available. The advances in this area will be discussed in light of the recent data, with structural and functional model compounds of N 2 OR active site.authorsversionpublishe

Enzymatic activity mastered by altering metal coordination spheres

J Biol Inorg Chem (2008) 13:1185–1195 DOI 10.1007/s00775-008-0414-3Metalloenzymes control enzymatic activity by changing the characteristics of the metal centers where catalysis takes place. The conversion between inactive and active states can be tuned by altering the coordination number of the metal site, and in some cases by an associated conformational change. These processes will be illustrated using heme proteins (cytochrome c nitrite reductase, cytochrome c peroxidase and cytochrome cd1 nitrite reductase), non-heme proteins (superoxide reductase and [NiFe]-hydrogenase), and copper proteins (nitrite and nitrous oxide reductases) as examples. These examples catalyze electron transfer reactions that include atom transfer, abstraction and insertion

Multivalent enzymes that enable the use of hydrogen peroxide for microaerobic and anaerobic proliferation

This work was financed by national funds from FCT - Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences - UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy - i4HB. FCT supported SRP through the projects PTDC/BIA-PRO/109796/2009 and PTDC/BIA-BQM/29442/2017, DSB and RNSO through the scholarships UI/BD/151168/2021, and “Verão com Ciência2020”, respectively.Bacterial peroxidases are responsible for the reduction of hydrogen peroxide to water. Found in the periplasm of gram-negative bacteria, they are one of the defense mechanisms against endogenous and exogenous peroxide stress under low oxygen tensions. Besides being involved in peroxide detoxification, bacterial peroxidases have been proposed to constitute an alternative pathway to the respiratory chain under anoxic conditions, as demonstrated in E. coli that can use hydrogen peroxide as an electron acceptor in the absence of oxygen. Bacterial peroxidases are c-type cytochromes with either two or three c-type hemes bound to the polypeptide chain, being divided into classical or non-classical, respectively. Orthologous to the classical bacterial peroxidases are the MauG enzymes that share some structural, spectroscopic and sequence similarities but have distinct physiological roles (though for most their function remains unknown). The spectroscopic and kinetic data on bacterial peroxidases are reviewed for both classes. Most classical bacterial peroxidases require reductive activation that consists in structural changes so that the catalytic heme becomes accessible to the substrate. However, non-classical enzymes are ready to bind the hydrogen peroxide as their catalytic center is penta-coordinated, which is also observed in their structural model. The few studies that report the involvement of bacterial peroxidases from pathogenic bacteria in biofilms, is an indication that these enzymes might contribute to their infection mechanism and thus can constitute alternative drug targetspublishersversionpublishe

Avaliação de competências da escrita em crianças do pré-escolar na faixa etária dos 4/5 anos

Relatório de Estágio apresentado à Escola Superior de Educação do Instituto Politécnico de Castelo Branco para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Educação Pré-Escolar e Ensino do Primeiro Ciclo do Ensino Básico.O presente relatório de estágio foi redigido na etapa final do Mestrado em Educação Pré-Escolar e Primeiro Ciclo do Ensino Básico, da Escola Superior de Educação de Castelo Branco, e intitula-se “Avaliação de competências da escrita em crianças do Pré-Escolar, na faixa etária dos 4/5 anos”. Neste relatório começamos por apresentar uma panorâmica geral do trabalho desenvolvido durante o Estágio Supervisionado em Educação Pré-Escolar, dado que a investigação empírica realizada decorreu neste mesmo contexto. Fazemos, pois, uma síntese da nossa atuação pedagógica ao longo das unidades didáticas que lecionámos individualmente, incidindo, em particular, na organização das atividades levadas a cabo. Na segunda parte do nosso relatório de estágio procedemos à descrição da investigação empírica que fizemos. Esta incidiu sobre o desenvolvimento de competências de escrita em crianças entre os quatro e cinco anos de idade, sendo que os objetivos específicos do estudo foram: avaliar a capacidade de diferenciação entre desenho e escrita; avaliar o conhecimento sobre a orientação da escrita; avaliar o conhecimento de letras; avaliar as conceptualizações das crianças sobre a escrita. Tratou-se de um tipo de investigação qualitativa, especificamente, um estudo de caso, com uma amostra de conveniência, constituída por um grupo de seis crianças. As crianças da amostra foram previamente referenciadas pela Educadora Cooperante como sendo, dentro da faixa etária dos 4/5 anos, as três mais e as três menos desenvolvidas da sala, em termos de linguagem escrita. A recolha de dados foi feita através da nossa própria observação, de uma entrevista à Educadora, de notas de campo, de registos fotográficos e de produções textuais das crianças. Para atingir os objetivos específicos definidos para o estudo, foram desenvolvidas quatro atividades sobre escrita, uma para avaliar a capacidade de diferenciação entre desenho e escrita, outra para avaliar o conhecimento sobre a orientação de escrita, uma outra para avaliar o conhecimento de letras uma última para avaliar as conceptualizações das crianças sobre escrita. Os resultados destas atividades constituíram a fonte de dados principal a tomar em consideração neste estudo. Quer a componente de Estágio Supervisionado deste mestrado quer a componente investigativa no terreno foram fundamentais para o nosso crescimento profissional, ajudando-nos a refletir de forma mais consistente sobre as competências que um profissional desta área deve desenvolver.Abstract - The present research report is included on the final stage of the Pre-School and Elementary School Education master’s degree, from the Castelo Branco University and it’s named “Writing Competence Evaluation on Pre-School Children Between the Ages of 4 and 5 years old”. This report begins by presenting a general idea of the work developed during the Pre-School education internship supervision, once the actual investigation was realized on the same context. So, we make a synthesis of our pedagogical work during the units we taught individually, focusing the description in the organization of the activities that we actually made. In the second part of our report we proceed to the description of the investigation. This part was focused on the development of the writing competence in children between the ages of four a five years old, once that the specific objectives of the study were: evaluate the capacity to differentiate the writing from the drawing; evaluate the writing orientation knowledge, evaluate the letters knowledge; evaluate the children’s conceptualizations about the writing. This investigation was strictly qualitative, specifically a case study involving a group of six children as a sample frame. The subjects from the sampling frame were previously referenced by the cooperating teacher by being between the ages of 4 and 5 and being the 3 less and 3 more developed children in the context of the writing language. The data collection was made through our own observation, by interviewing the teacher, from field notes, from photographic records and from children textual productions. To accomplished the specific defined goals of the study, were developed four activities involving writing, the first one to evaluate the capacity to differentiate the writing from the drawing; the second one to evaluate the writing orientation knowledge, the third one to evaluate the letters knowledge and the last one to evaluate the children´s conceptualizations about the writing. The results of this activities as became a source of central data to take under consideration on this study. Either the internship supervision component of these master´s degree or the field investigation component were both fundamentals for our professional growth, helping us to reflect in a more consciously way about the skills that a professional in this field should develop

Proton-coupled electron transfer mechanisms of the copper centres of nitrous oxide reductase from Marinobacter hydrocarbonoclasticus – An electrochemical study

PTDC/BBB-BQB/0129/2014, FCT/MCTES, UID/Multi/04378/2019, UID/QUI/50006/2019, Centro de Quimica Estrutural multiannual funding 2020-2023, UID/QUI/00100/2019.Reduction of N2O to N2 is catalysed by nitrous oxide reductase in the last step of the denitrification pathway. This multicopper enzyme has an electron transferring centre, CuA, and a tetranuclear copper-sulfide catalytic centre, “CuZ”, which exists as CuZ*(4Cu1S) or CuZ(4Cu2S). The redox behaviour of these metal centres in Marinobacter hydrocarbonoclasticus nitrous oxide reductase was investigated by potentiometry and for the first time by direct electrochemistry. The reduction potential of CuA and CuZ(4Cu2S) was estimated by potentiometry to be +275 ± 5 mV and +65 ± 5 mV vs SHE, respectively, at pH 7.6. A proton-coupled electron transfer mechanism governs CuZ(4Cu2S) reduction potential, due to the protonation/deprotonation of Lys397 with a pKox of 6.0 ± 0.1 and a pKred of 9.2 ± 0.1. The reduction potential of CuA, in enzyme samples with CuZ*(4Cu1S), is controlled by protonation of the coordinating histidine residues in a two-proton coupled electron transfer process. In the cyclic voltammograms, two redox pairs were identified corresponding to CuA and CuZ(4Cu2S), with no additional signals being detected that could be attributed to CuZ*(4Cu1S). However, an enhanced cathodic signal for the activated enzyme was observed under turnover conditions, which is explained by the binding of nitrous oxide to CuZ0(4Cu1S), an intermediate species in the catalytic cycle.authorsversionpublishe