Structural and Functional Characterization of a New Bacterial Target Against Tuberculosis: The Phosphatase PtpA

Tuberculosis (TB) is one of the top causes of death remaining a major public health problem worldwide. Mycobacterium tuberculosis is the agent of TB, infecting the human respiratory tract. Its remarkable pathogenicity hinges upon the ability to challenge the immune system of the host by secreting phosphatases into macrophages. Among them, Protein Tyrosine Phosphatase A (PtpA) plays a key role on the infection process, preventing the phagosome-lysosome fusion and promoting the inhibition of phagosome acidification. Thus, PtpA becomes a promising target for the development of new anti-TB drugs. The aim of this work is to contribute to find new structure-based drug design approaches against TB, studying the inhibitory properties of three different families of compounds towards PtpA – chalcones, thiosemicarbazones and azaindoles. The protein was overexpressed in E. coli – final yield of 20 mg protein/ liter of culture – and successfully purified using affinity chromatography. To provide new insights into the binding mode of the studied compounds, molecular docking studies were performed suggesting thiosemicarbazones as non-competitive inhibitors and the chalcones and azaindoles with a preferential active site binding. The protein was also biophysically characterized. The oligomeric state was confirmed by SEC, proving that PtpA is a monomer in solution. The protein stability was assessed through TSA revealing that, with 10% glycerol, PtpA resists to the effects of 10% DMSO. TSA was also used to find a suitable protein storage condition (-80°C) and to confirm PEG400 as an alternative solvent for the inhibitors. In addition, distinct biophysical approaches – TSA, MST and urea-gel electrophoresis – were implemented to detect protein-ligand interactions but definitive evidence were not obtained. Ligand-free and co-crystallization assays were extensively explored and several crystals were tested at the ESRF, Diamond and MAX IV. Two crystal structures were obtained: a co-crystallization PtpA-Lap11 structure at 3.6 Å resolution and a soaking PtpA-C33 structure at 2.8 Å resolution. Despite the low/medium resolution obtained, both structures reveal the potential binding of the inhibitors with suspicious density blobs near His120B for Lap11 and at the active site for C33. The ligands were preliminarily modelled but further refinement cycles are required to elucidate the respective binding

Estudo sobre as diferenças interativas e comunicativas dos educadores e dos pais com crianças em idade pré-escolar

A investigação sobre a qualidade das interações, a comunicação e a atividade entre crianças e adultos tem sido desenvolvida essencialmente com figuras femininas, continuando por esclarecer o papel dos homens como pares afetivos e promotores do desenvolvimento e bem-estar da criança. Procurámos estudar o comportamento de educadores e de pais, do género masculino, com crianças entre os 3 e os 5 anos, sem problemas de desenvolvimento. Neste estudo, 10 educadores (com uma menina e com um menino) e 17 pais (com o seu filho ou filha) foram observados independentemente, na mesma situação semi-experimental. Foi-lhes pedido que realizassem, em 20 minutos, um produto, à sua escolha, com materiais e ferramentas disponibilizadas. Pretendia-se descrever e comparar os dois grupos quanto: i) aos produtos realizados; e uso de materiais e ferramentas; ii) à empatia, atenção, reciprocidade, cooperação, elaboração/fantasia e desafio proposto; e iii) à qualidade da comunicação verbal. O estudo indica que existem estilos de atividade e cooperação distintos nos dois grupos estudados. Comparativamente aos pais, os educadores estimularam mais a exploração de novos problemas e conceitos. Nas díades pai-filho(a) foram elaborados produtos com mais componentes e utilizadas mais ferramentas. Adicionalmente, os pais fizeram mais perguntas de processo e elogios do que os educadores que, por seu lado, realizaram mais perguntas de conteudo sugestões e críticas. À luz da investigação anterior, refletimos sobre os papéis das figuras masculinas em educação de infância e na família; e o seu contributo para o desenvolvimento da criança.Abstract The previous research about the quality of interactions, communication and activity among children and adults has essentially been developed with female figures, continuing to clarify the role of men as affective figures and in promoting child development and welfare. The aim of this study is to compare male educators and fathers’ behavior with children between 3 and 5 years without developing problems. In this study, 10 male educators (with a girl and a boy) and 17 fathers (with the infant) have been observed independently in the same semi-experimental play situation. For this purpose, they were asked to manufacture a product of their choice with the materials and tools available, in 20 minutes. This study aims are to: i) describe and compare products made by fathers and male educators, as well as the choice of materials; ii) compare the interactive behavior of male and female educators regarding empathy, ability to challenge the child, dialogical interaction, communication contends and cooperativity with child; and iii) the quality of communication. Our findings indicate differences in the way male educators and fathers communicate and interact with children. Comparing with parents, children work with more autonomy, elaborated products with less elements (mostly subjects), and used less tools with educators. Furthermore, fathers made more process questions and positive comments while educators made more content questions, suggestions and negative remarks. Taking past studies, we reflect about the roles of male figures in childhood education and fathers’ contribution to the child development.Résumé La recherche sur la qualité des interactions, de la communication et de l'activité entre les enfants et les adultes a été développée surtout avec des figures féminines, n´étant pas encore clarifié le rôle des hommes comme pairs affectifs et promoteurs du développement et du bien-être des enfants. Ces constats nous ont incités à observer de plus près le comportement des éducateurs et des pères d'enfants âgés de 3 à 5 ans, sans problèmes de développement. Dans cette étude, nous avons observé indépendamment, dans la même situation semi-expérimentale, 10 éducateurs (avec une fille et un garçon) et 17 pères (avec leur fils ou leur fille). Ils ont été invités à fabriquer, en 20 minutes, un produit, à leur choix, avec les matériaux et les outils disponibles. Il était prévu de comparer les deux groupes en ce qui concerne: i) les artefacts produits; l’utilisation de matériaux et d'outils; ii) l’empathie, l’attention, la réciprocité, la coopération, l’élaboration / fantaisie et le défi vécu; et iii) la qualité de la communication verbale. Cette recherche met en évidence qu'il existe différents styles d'activité et de coopération dans les deux groupes étudiés: les éducateurs ont stimulé davantage l'exploration de nouveaux problèmes et concepts; les dyades des pères ont élaboré des produits avec plus de composants, en utilisant plus d'outils; les pères ont posé plus de questions procédurales et ont davantage félicité que les éducateurs qui, à leur tour, ont posé plus de questions sur le contenu, ont fait plus de suggestions et de critiques. En consonance avec des études préalables, nous réfléchissons sur le rôle des figures masculines dans l'éducation des enfants et dans la famille; et leur contribution au développement de l'enfant.info:eu-repo/semantics/publishedVersio

Estudo sobre as diferenças interativas e comunicativas das educadoras e das mães com crianças em idade pré-escolar

Os educadores de infância, na ausência dos pais, asseguram cuidados e garantem o bem estar da criança assim como a sua educação e desenvolvimento. A criança estabelece com a maioria dos educadores uma relação afetiva privilegiada mas não de vinculação. O estudo das diferenças e semelhanças entre estes dois tipos de relação permite averiguar a diversidade de relações proporcionadas à criança e o contributo da formação profissional das educadoras no estabelecimento da relação com as crianças. Neste estudo, mães e educadoras são observadas independentemente, na mesma situação quasi-experimental com a criança, como parceiros numa atividade lúdica de construção, sendo analisada a qualidade interativa e comunicativa das díades. Para o efeito, foi pedido a 19 díades mãe-filho(a) e 22 díades educadora-criança que realizassem, em 20 minutos, um produto à sua escolha com os materiais e ferramentas disponibilizados. As crianças tinham entre 3 e 5 anos e não apresentavam problemas de desenvolvimento. Pretendia-se: 1) descrever e comparar os produtos realizados pelas díades mãe-criança e educadora-criança, bem descrever como as escolhas de materiais; 2) comparar a qualidade interativa das mães e das educadoras quanto à empatia, atenção, reciprocidade, cooperação, elaboração/fantasia e desafio proposto; e 3) a qualidade da comunicação quanto ao uso de sugestões, perguntas, ordens, elogios e críticas. Os dados indicam que as educadoras deram mais espaço para a atuação da criança, enquanto as mães procuraram cumprir o tempo da tarefa e aprimorar o produto com as crianças. As mães usaram o reforço verbal para organizar a atividade, impor limites/regras e motivar a criança, enquanto as educadoras recorreram à comunicação oral para transmitir conteúdos e dirigir a atividade.Abstract The educators, in the absence of parents, provide care and ensure the welfare of the child as well as their education and development. Educators are children significant emotional figures but parents are attachment figures. The study of behavioral differences and similarities between educators and mothers can contribute to our knowledge about these two types of relationships and to observe how the educators training can contribute for their interaction with the children. In this study, mothers and educators are observed independently in the same semi experimental situation with the child, as partners in a playful activity of construction, and the interactive and communicative quality of dyads is analyzed. For this purpose, 19 dyads mother-child and 22 teacher-child dyads were asked to make in 20 minutes a product of their choice with the materials and tools available. The children were between 3 and 5 years old without developing problems. The aims of this study was to: 1) describe and compare the products made by the mother and educator dyads’, as well as their choice of materials; 2) compare the quality of mothers and educators’ interaction as well as their empathy, attention, reciprocity, cooperation, development/fantasy and proposed challenge; and 3) the quality of communication as well as suggestions, questions, directions and positive or negative feedback. Our findings indicate that the educators allow and support the child's actions while mothers were more focused on compliance with the time and product improvement. Mothers used the verbal reinforcement to organize the activity, to set limits/rules and to motivate the child while the educators relied on verbal communication to introduce contents and organize the activity.Résumé La recherche sur la qualité des interactions, la communication et l’activité entre enfants et adultes, s’est développé en grande partie autour des figures féminines, restant à clarifier le rôle des hommes comme partenaires affectives et promoteurs du développement et bien-être de l’enfant. Nous avons voulu étudier le comportement des éducateurs et des pères avec des enfants âgés de 3 à 5 ans, sans problèmes de développement. En cette étude, 10 éducateurs (avec une fille et avec un garçon) et 17 pères (avec son fils ou sa fille) ont été observés indépendamment, dans la même situation semi-expérimentale. Nous leur avons demandé de réaliser en 20 minutes un produit, à sa guise, avec les matériels et les outils mis à leur disposition. Nous prétendions une description et comparaison entre les deux groupes relative : i) aux produits réalisés et les matériels et outils utilisés ; ii) à l’empathie, attention, réciprocité, coopération, élaboration/fantaisie et défi proposé ; et iii) à la qualité de la communication verbal. L’étude indique l’existence de styles d’activité et coopération distinctes dans les deux groupes étudiés. Comparativement aux pères, les éducateurs ont stimulé davantage l’exploration de nouveaux problèmes et concepts. Dans les dyades père-enfant les produits avaient été réalisés faisant recours à plus d’outils et ils avaient plus d’éléments. En plus, les pères ont émis plus des questions de processus et des éloges comparativement aux éducateurs qui, a leur tour, ont émis plus des questions de contenu, suggestions et critiques. À la lumière de la recherche antérieure, nous avons réfléchi sur les rôles des figures masculines dans l’éducation de la petite enfance et dans la famille, et son apport au développement de l’enfant.info:eu-repo/semantics/publishedVersio

Estudo comparativo acerca do comportamento e comunicação materna e paterna em atividade conjunta com os seus filhos de idade pré-escolar

The previous research performed in children life contexts tend to describe fathers as non-directive playmates; while mothers tend to reinforce the communication during interactions and involve affectively the child. When mothers are playmates and fathers give basic care to their child the quality of the interaction of mother-infant and father-infant increases considerably. In this study, fathers and mothers were observed independently in the same semi-experimental play situation with their children. For this purpose, 19 dyads mother-infant and 17 dyads father-infant participated in this 20 minutes experience, to manufacture one product using the materials and tools available to one’s choice. Children were between 3 and 5 years old and had no developmental problems identified. We aimed to: (i) compare the interactive quality of parents related to empathy, reciprocity, cooperation, fantasy and challenge; (ii) the communication quality and (iii) describe and compare the products made by the dyads mother-infant and father-infant, as well as the chosen materials. Our findings indicate few differences between mothers and fathers behaviour. In terms of communication, mothers communicate more with process questions than parents. The major differences correspond to how parents react to boys and girls, giving girls more freedom to explore, more positive feedback, and being more responsive to their emotions. The boys lost more often interest during the activity than the girls. Children’s gender affected the results more than the parents’, which indicates that the parents interact and communicate distinctly with girls and boys. Additionally, parents’ educational level correlated with more attentive, patient, and cooperative behaviour with their children. Regarding the age of the parents: younger parents and with more children use more materials and tools.A investigação tende a descrever o pai como parceiro de jogo que favorece a liberdade de ação; enquanto a mãe, tende a reforçar a comunicação nas interações, envolvendo afetivamente a criança. Nos casos em que a mãe brinca e o pai presta cuidados básicos à criança, a qualidade da interação pai-filhos aumenta consideravelmente. Neste estudo, observamos pai e mãe independentemente na mesma situação experimental como parceiros da criança numa atividade lúdica de construção. Comparamos os seus comportamentos quando colocados no mesmo papel. Para o efeito, foi pedido a 19 díades mãe-filho(a) e 17 díades pai-filho(a) que realizassem, em 20 minutos, um produto à sua escolha com os materiais e ferramentas disponibilizados. As crianças tinham entre 3 e 5 anos, sem atrasos de desenvolvimento identificados. Pretendemos descrever e comparar os pais (mães e pais considerados em conjunto) quanto: (i) à empatia, atenção, reciprocidade, cooperação, elaboração/fantasia e desafio proposto; (ii) à qualidade da comunicação; e (iii) aos produtos realizados e escolhas de materiais. Os nossos resultados indicam poucas diferenças entre pais e mães. Em termos de comunicação, as mães realizam mais perguntas de processo do que os pais. As diferenças mais relevantes correspondem à forma como os pais e as mães reagem com os meninos e as meninas, dando maior liberdade de ação às meninas, mais feedback positivo e revelando-se mais sensíveis a responder às suas emoções. Os meninos perderam mais interesse durante a atividade e revelaram mais sinais de aborrecimento do que as meninas. O sexo das crianças afetou mais os resultados do que o dos pais, ou seja, os pais interagiram e comunicaram distintamente com meninas e meninos. Adicionalmente, a escolaridade dos pais correlacionou-se com comportamentos mais atentos, pacientes e cooperativos dos pais. Relativamente à idade dos progenitores, os pais mais novos e com mais filhos usaram mais materiais e ferramentas

Estudo sobre as diferenças interativas e comunicativas de educadores e educadoras com crianças em idade em pré-escolar

A educação de infância tem sido assegurada maioritariamente por mulheres e associada ao género feminino. Recentemente, o número de homens na profissão aumentou na Europa. Contudo, o contributo do educador masculino tem sido pouco estudado. No intuito de contribuir para esse corpo de conhecimento, procurámos estudar as diferenças e semelhanças nos comportamentos interativos dos educadores e das educadoras com crianças de 3 anos sem problemas de desenvolvimento. Neste estudo, educadores e educadoras foram observados independentemente na mesma situação experimental como parceiros numa atividade lúdica de construção, sendo analisada a qualidade interativa e comunicativa dos Educadores. Para o efeito, foi pedido a 10 educadores e 11 educadoras que realizassem, em 20 minutos, um produto com as crianças com os materiais e ferramentas disponibilizadas à escolha de ambos. Pretendia-se: i) descrever e comparar os produtos realizados pelas díades masculinas e femininas, bem como as escolhas de materiais; ii) comparar a qualidade interativa dos educadores e das educadoras quanto à empatia, atenção, reciprocidade, cooperação, elaboração/fantasia e desafio proposto; e iii) a qualidade da comunicação. Os dados parecem indicar que existem algumas diferenças na forma de comunicar das educadoras e dos educadores mas poucas diferenças na qualidade interativa. A experiência profissional, a situação de emprego e o facto de serem ou não pais condicionou o comportamento dos Educadores.Abstract Early childhood education has been ensured mostly by women, and this profession is traditionally associated with the feminine gender. Recently, the number of men in this profession has increased in Europe. However, the role of the male educator is understudied. In order to contribute to this body of knowledge, we studied the differences and similarities of the interactive behavior of male and female educators with children with 3 years old without developmental problems. In this study, female and male educators were observed independently in the same experimental situation as partners with the child in a playful activity of construction. Later, we analyzed the interactive and communicative quality of Educators. For that purpose, 10 male educators and 11 female educators built a product of their choice, with the child, with the materials and tools available in about 20 minutes. This study aims were to: i) describe and compare products made by male and female dyads, as well as the choice of materials; ii) compare the interactive behavior of male and female educators regarding empathy, ability to challenge the child, dialogical interaction, communication contends and cooperativity with child; and iii) the quality of communication. Our findings indicate differences in the way male and female educators communicate but fewer differences in the interactive quality. The professional experience, employment status and parenthood seemed to affect their behavior.info:eu-repo/semantics/publishedVersio

Arbustus unedo essence: morphological and genetic characterization of the strawberry tree of Castelo de Paiva

O medronheiro é um arbusto da região mediterrânica que pode ser encontrada por todo o país. Ao contrário do que verifica na região sul do país, no concelho de Castelo de Paiva é atribuída uma reduzida importância económica a esta espécie. Com o intuito de preservar e potenciar a produção desta espécie e contribuir para a dinamização da economia do concelho, procedeu-se à caracterização morfológica e genética de uma amostra da população de medronheiros de Castelo de Paiva. A caracterização morfológica e genética foi realizada para um total de 10 genótipos. Para tal recolheram-se 70 folhas aleatoriamente em cada árvore. Em 40 folhas mediu-se o comprimento, largura, comprimento do pedúnculo, peso fresco, peso seco e determinou-se a área foliar. Dos caracteres morfológicos analisados, aqueles que se revelaram mais úteis na distinção dos vários genótipos foram: comprimento do pedúnculo, peso fresco e peso seco. As restantes 30 folhas foram utilizadas para a caracterização genética. Esta caracterização foi realizada recorrendo a um marcador de DNA, ISSR. Os 5 primeiros exemplaresutilizados na técnica de ISSR demonstraram-se polimórficos. Os resultados da caracterização genética sugerem que a variabilidade genética na população é média a alta.The strawberry tree is a shrub native in the Mediterranean region and it can be found throughout Portugal. Unlike the case in the southern region of the country, in Castelo de Paiva a minor economic importance is given to this species. In order to preserve, to enhance the production of this species and to contribute to the boosting of the economy of the region, we proceeded to the characterization of a small sample population of this fruit tree of Castelo de Paiva in what concerns to its morphology and genetics. The morphological and genetic characterization was performed for a total of 10 genotypes. For this, 70 leaves were randomly collected from each tree. For 40 leaves, it was measured the length, the width, the peduncle length, the wet weight, the dry weight and determined the leaf area. Of the morphological characteristics analyzed, the ones that proved most useful in distinguishing the various genotypes were: the length peduncle, the wet weight and the dry weight. The remaining 30 leaves were used in the genetic characterization. This characterization was performed using a DNA marker, the ISSR. The 5 primers used in the ISSR technique proved to be polymorphic. The results from the genetic characterization suggest that variability in population genetics is medium to high

Risk factors for infection, predictors of severe disease, and antibody response to COVID-19 in patients with inflammatory rheumatic diseases in Portugal: a multicenter, nationwide study

Copyright © 2022 Cruz-Machado, Barreira, Bandeira, Veldhoen, Gomes, Serrano, Duarte, Rato, Miguel Fernandes, Garcia, Pinheiro, Bernardes, Madeira, Miguel, Torres, Bento Silva, Pestana, Almeida, Mazeda, Cunha Santos, Pinto, Sousa, Parente, Sequeira, Santos, Fonseca and Romão. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Objective: To identify risk factors for SARS-CoV-2 infection and for severe/critical COVID-19, and to assess the humoral response after COVID-19 in these patients. Methods: Nationwide study of adult patients with inflammatory RMDs prospectively followed in the Rheumatic Diseases Portuguese Register-Reuma.pt-during the first 6 months of the pandemic. We compared patients with COVID-19 with those who did not develop the disease and patients with mild/moderate disease with those exhibiting severe/critical COVID-19. IgG antibodies against SARS-CoV-2 were measured ≥3 months after infection and results were compared with matched controls. Results: 162 cases of COVID-19 were registered in a total of 6,363 appointments. Patients treated with TNF inhibitors (TNFi; OR = 0.160, 95% CI 0.099-0.260, P < 0.001) and tocilizumab (OR 0.147, 95% CI 0.053-0.408, P < 0.001) had reduced odds of infection. Further, TNFi tended to be protective of severe and critical disease. Older age, major comorbidities, and rituximab were associated with an increased risk of infection and worse prognosis. Most patients with inflammatory RMDs (86.2%) developed a robust antibody response. Seroconversion was associated with symptomatic disease (OR 13.46, 95% CI 2.21-81.85, P = 0.005) and tended to be blunted by TNFi (OR 0.17, 95% CI 0.03-1.05; P = 0.057). Conclusions: TNFi and tocilizumab reduced the risk of infection by SARS-CoV-2. Treatment with TNFi also tended to reduce rates of severe disease and seroconversion. Older age, general comorbidities and rituximab were associated with increased risk for infection and worse prognosis, in line with previous reports. Most patients with RMDs developed a proper antibody response after COVID-19, particularly if they had symptomatic disease.We acknowledge the generous sharing of the expression constructs by Dr. Florian Krammer, Icahn School of Medicine at Mount Sinai, New York, USA [Development of SARS-CoV-2 reagents was partially supported by the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C] and the protein production by Drs. Paula Alves and Rute Castro at Instituto de Biologia Experimental e Tecnológica (iBET) Oeiras, Portugal as part of the Serology COVID consortium.info:eu-repo/semantics/publishedVersio

EGF functionalized polymer-coated gold nanoparticles promote EGF photostability and EGFR internalization for photothermal therapy

The application of functionalized nanocarriers on photothermal therapy for cancer ablation has wide interest. The success of this application depends on the therapeutic efficiency and biocompatibility of the system, but also on the stability and biorecognition of the conjugated protein. This study aims at investigating the hypothesis that EGF functionalized polymer -coated gold nanoparticles promote EGF photostability and EGFR internalization, making these conjugated particles suitable for photothermal therapy. The conjugated gold nanoparticles (100-200 nm) showed a plasmon absorption band located within the near infrared range (650-900 nm), optimal for photothermal therapy applications. The effects of temperature, of polymer-coated gold nanoparticles and of UVB light (295nm) on the fluorescence properties of EGF have been investigated with steady-state and time-resolved fluorescence spectroscopy. The fluorescence properties of EGF, including the formation of Trp and Tyr photoproducts, is modulated by temperature and by the intensity of the excitation light. The presence of polymeric-coated gold nanoparticles reduced or even avoided the formation of Trp and Tyr photoproducts when EGF is exposed to UVB light, protecting this way the structure and function of EGF. Cytotoxicity studies of conjugated nanoparticles carried out in normal-like human keratinocytes showed small, concentration dependent decreases in cell viability (0-25%). Moreover, conjugated nanoparticles could activate and induce the internalization of overexpressed Epidermal Growth Factor Receptor in human lung carcinoma cells. In conclusion, the gold nanoparticles conjugated with Epidermal Growth Factor and coated with biopolymers developed in this work, show a potential application for near infrared photothermal therapy, which may efficiently destroy solid tumours, reducing the damage of the healthy tissue.Support was provided by: Fundacao para a Ciencia e Tecnologia (FCT) for the financial support under the project reference PTDC/BBB-BMC/0611/2012 [https://www.fct.pt/apoios/projectos)]. The work at CBMA was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalizacao (POCI) [https://www.fct.pt/apoios/projectos]; European Commission through the project H2020-644242-SAPHELY (https://saphely.eu/project.php) and the project H2020-634013-2-PHOCNOSIS [http://cordis.europa.eu/project/rcn/193268_en.html].The authors would like to thank Fundacao para a Ciencia e Tecnologia (FCT) for the financial support under the project reference PTDC/BBB-BMC/0611/2012. The work at CBMA was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalizacao (POCI). The authors acknowledge the funding from the European Commission through the project H2020-644242-SAPHELY and the project H2020-634013-2-PHOCNOSIS. Finally, the authors would also like to thank the master student Joao Lopes from Universidade Lusofona (Portugal) for the help with in vitro cytotoxic assays. Isabel Correia acknowledges FCT for Investigator FCT contract.info:eu-repo/semantics/publishedVersio

Brincar a dois tempos: estilos de comunicação entre Pais e Filhos em jogo livre

A investigação no campo do desenvolvimento infantil, partindo de modelos conceptuais como a teoria da vinculação (Bowlby, 1969) ou modelos e teorias de aprendizagem (e.g., Bandura, 1963), acabou por se orientar em dois vetores: cognitivo e relacional. Carece, ainda, a literatura científica de trabalhos que procurem compreender como estes domínios se suportam e afetam mutuamente. Os trabalhos de António Damásio (2013) indicam que não existem decisões estritamente emocionais ou cognitivas e Crittenden (1995) inclui essas dimensões na no seu modelo de maturação dinâmica da vinculação (do verdadeiro ao falso afeto, da verdadeira à falsa cognição). Neste estudo procurámos, numa atividade orientada para tarefa, analisar os domínios cognitivos (através do jogo) e afetivo e a sua inter-relação assumindo uma perspetiva marcadamente diádica. Procurando, simultaneamente, em cada parceiro os reflexos das intenções comunicativas do outro. A nossa análise identifica condições molares (afetividade e jogo) na interação diádica a partir de indicadores de nível molecular. Neste nível molecular a dimensão relacional agrupa a resposta vocal, facial e as trocas afetivas; o jogo: a reciprocidade, a atividade lúdica e a diretividade.info:eu-repo/semantics/publishedVersio

UV-light power effect (different excitation slit openings) on EGF photochemistry: A) EGF fluorescence excitation and emission wavelengths were fixed at 295 nm and 330 nm, respectively, for 0.1 mm (0.12 μW), 0.5 mm (0.30 μW), 0.8 mm (1.67 μW), 1.2 mm (2.34 μW) and 2.0 mm (4.40 μW); B) SYPRO^® Orange fluorescence excitation and emission wavelengths were fixed at 470 nm and 580 nm, for the same power levels.

<p>Continuous illumination was conducted during 2 hours and the temperature of each solution was kept at 20°C for all experiments.</p