The role of α2,3- and α2,6-sialylation

RESUMO: Os glicoconjugados que decoram a superfície celular e os lípidos e proteínas secretados ocupam o ponto de encontro onde normalmente ocorrem interacções críticas homólogas (hospedeiro-hospedeiro) e heterólogas (hospedeiro-patogénio). Apesar de ser largamente aceite que os glicanos são parte integrante do processo de imunidade, continua a não ser claro qual o papel que os glicanos, em toda a sua diversidade, tomam no quadro geral da imunidade. Os glicanos, que são frequentemente terminados por resíduos de ácido siálico, podem ser alterados por factores externos, tais como patogénios, ou por acontecimentos fisiológicos celulares específicos. Normalmente em posição terminal, as glico-estruturas que contêm ácido siálico assumem um papel fundamental numa quantidade substancial de receptores imunes envolvidos na adesividade e tráfico celular, tal como as Selectinas e as Siglecs, das quais se sabe apresentarem uma relevante função imune. À altura do início desta tese, era sabido que os ácidos siálicos expressos à superfície das células poderiam modular mecanismos importantes nas respostas imunes adaptativas. Considerando a posição de charneira que as células dendríticas (DCs) ocupam na transição da resposta imune inata para a adaptativa, antecipámos que os ácidos siálicos poderiam também modular mecanismos relevantes nas DCs humanas. As DCs têm uma função muito relevante na verificação e captura antigénica, migração para os gânglios linfáticos e apresentação antigénica aos linfócitos, uma sequência de funções que conduz, em ultima instância, à indução da resposta inata adaptativa. Considerando estas premissas, a nossa hipótese principal foi que os ácidos siálicos podem influenciar funções relevantes das DCs, tais como captura de antigénios, maturação, migração para os gânglios linfáticos e apresentação antigénica às células Para testar esta hipótese, dividimos o trabalho em quatro partes: 1) Analisámos os glicanos sialilados de superfície, expressos durante a diferenciação de monócitos humanos em DCs (moDCs). Os nossos dados mostraram que a expressão dos glicanos com ligações em O (O-glicanos) e sialilados em α2,3, assim como glicanos com ligações em N (N-glicanos) sialilados em α2,6 e α2,3 aumentou durante o processo de diferenciação das moDCs. Contribuindo para esta nova configuração glicosídica, três sialiltransferases (STs) poderão estar envolvidas: a ST6Gal-1 correlaciona-se com a expressão aumentada de N-glicanos sialilados em α2,6; a ST3Gal-1 contribui para a sialilação em α2,3 de O-glicanos, em especial de antigénios T; e a ST3Gal-4 poderá ser responsável pelo aumento de N-glicanos sialilados em α2,3. Após estímulo e consequente maturação das moDCs, ambos os níveis de expressão génica de ST6Gal-1 e ST3Gal-4 são negativamente modificados sendo, também, que a expressão de ST3Gal-1 varia consoante o estímulo. 2) Estudámos posteriormente as consequências da modulação dos ácidos siálicos de superfície nas funções das DCs. Observámos que a remoção dos ácidos siálicos de superfície diminui significativamente a capacidade de macropinocitose e endocitose mediada por receptores nas moDCs. Em contrapartida, o tratamento com sialidase aumentou significativamente a capacidade das moDCs para fagocitar Escherichia coli. Determinou-se também que este mecanismo requer a existência de ácido siálico presente nas E. coli indicando um mecanismo de interacção hospedeiro-patogénio dependente de ácido siálico em ambas as partes envolvidas. As moDCs tratadas com sialidase também apresentam um nível superior de expressão de moléculas de MHC e moléculas co-estimulatórias, sugerindo um fenótipo celular mais maduro. Recorrendo ao modelo de ratinho, utilizaram-se DCs derivadas de células da medula (BMDCs) de ratinhos deficientes em ST3Gal-1 e ST6Gal-1. Estes ensaios revelaram que quer a endocitose quer a maturação são influenciadas por modificações 37 nos glicanos sialilados em α2,3 ou α2,6. A detecção e quantificação de proteínas Nglicosiladas e sialiladas em α2,6 apontou para um potencial envolvimento de integrinas β2 nestes mecanismos. 3) O efeito da sialilação em α2,6 na migração das DCs para os gânglios linfáticos foi também analisado. Observámos que BMDCs deficientes para ST6Gal-1 apresentam uma redução de cerca de 50% nos níveis de migração das DCs para os gânglios linfáticos, tal como aferido em ensaios de inflamação in situ e estudos de transferência adoptiva de células. Uma redução dos níveis deste tipo de migração foi também observada quando BMDCs nativas foram transferidas para ratinhos receptores deficientes em ST6Gal-1. São, contudo, necessários mais ensaios de forma a identificar as moléculas envolvidas neste processo. 4) Por último, analisámos o impacto da sialilação na estimulação antigénica das DCs às células T. Assim, concluiu-se que moDCs tratadas com sialidase apresentam um nível de expressão superior de IL-12, TNF-ɑ, IL-6 e IL-10, e activação do factor de transcrição nuclear kappa B (NF-κB). As DCs tratadas com sialidase induziram uma maior proliferação nas células T, com expressão correspondente de interferão-γ. Este dado sugere que a remoção de ácidos siálicos de superfície contribui para o desenvolvimento de uma resposta pro-inflamatória do tipo 1 por células T auxiliares (resposta Th1). Considerando estes dados no seu todo, concluímos que o ácido siálico tem um papel marcante nas funções imunes das DCs. Alterações à concentração de ácido siálico à superfície das células podem alterar a endocitose/fagocitose, maturação, migração para os tecidos e gânglios linfáticos e capacidade estimulatória para com as células T. Complementando estes dados, as ligações glicosídicas de ácidos siálicos criados por ST6Gal-1 e ST3Gal-1 são funcionalmente relevantes. A modulação programada da sialilação do glicocálice, mediada por sialidases individuais ou sialiltransferases é uma possibilidade aceitável para a melhoria da fagocitose por DCs e da sua potência imunológica. Este facto tem um significado particular para imunoterapias baseadas em DCs, podendo provar-se decisivo para a sua eficiência e aplicabilidade num futuro muito próximo.-------------------------------ABSTRACT: Glycans decorating cell surface and secreted proteins and lipids occupy the junction where critical host–host and host-pathogen interactions occur. In spite of the wide acceptance that glycans are centrally implicated in immunity, exactly how glycans and their variety and variability contribute to the overall immune response remains poorly defined. Glycans, frequently terminated by sialic acid residues, may be modified by external factors such as pathogens or upon specific physiological cellular events. The terminal, privileged positions of sialic acid-modified structures makes them key, fundamental determinants for a number of immune receptors with known involvement in cellular adhesiveness and cell trafficking, such as Selectins and Siglecs, with known relevant immune functions. At the time this thesis was initiated, it was established that sialic acids expressed at cell surface could modulate important mechanisms of the adaptive immune responses. Given the key role of dendritic cells (DCs) in the transition from innate to the adaptive immune responses, we anticipated that sialic acids could also modulate important mechanisms of human DCs. DCs have a relevant role in antigen screening and uptake, migration to lymph nodes and antigen presentation to lymphocytes, ultimately triggering the adaptive immune response. Therefore, our primary hypothesis was that sialic acids may modulate DC functions, such as antigen uptake, maturation, homing to lymph nodes and antigen presentation to T cells. To test this hypothesis, we divided our work in four parts. 1) Surface sialylated glycans expressed during differentiation from human monocytes to DCs (moDCs) were analyzed. Our data showed that α2,3-sialylated O-glycans and α2,6- and α2,3-sialylated N-glycans expression increased during moDC differentiation. Three main sialyltransferases (STs) are committed with this new glycan configuration: ST6Gal- 1 correlates with the increased expression of α2,6-sialylated N-glycans; ST3Gal-1 32 contributes for the α2,3-sialylation of O-glycans, especially T antigens; and ST3Gal-4 may contribute for the increased α2,3-sialylated N-glycans. Upon moDC maturation, ST6Gal-1 and ST3Gal-4 are downregulated and ST3Gal-1 is altered in a stimulus dependent manner. 2) We subsequently analyzed the consequences of the modulation of cell surface sialic acids in DC functions. We observed that removing surface sialic acid by sialidase significantly decreased the capacity of moDCs to micropinocytose and receptormediated endocytose. In contrast, treatment with a sialidase significantly improved the capacity of moDCs to phagocytose Escherichia coli. The improved phagocytosis mechanism required E. coli sialic acids, indicating a mechanism of host–pathogen interaction dependent on sialic acid moieties. Sialidase-treated moDCs have increased expression of MHC and co-stimulatory molecules, suggesting a more mature phenotype. Experiments using mouse bone marrow-derived DCs (BMDCs) from ST3Gal-1-/- and ST6Gal-1-/- strains indicated that endocytosis and maturation are influenced by changes in either α2,3 or α2,6-sialylated glycans. The analysis of α2,6-sialylated, N-glycosylated proteins, strongly suggested the potential involvement of β2 integrins, underlying these mechanisms. 3) The effect of α2,6-sialylation in DC homing to lymph nodes was also analyzed. We observed that BMDCs deficient for ST6Gal-1 have an almost 50% reduction in DC homing, as assayed by in situ inflammation and adoptive transfer studies. A reduction in DC homing was also observed when wild type BMDCs were transferred into ST6Gal-1-/- recipient mice. Further investigations are necessary to identify the molecules involved in this process. 4) Finally, we also analyzed the impact of sialylation on DCs ability to prime T cells. Sialidase-treated moDCs show increased gene expression of IL-12, TNF-α, IL-6 and IL- 10 cytokines, and activation of the transcription factor nuclear factor-κB. Sialidase33 treated DCs induced a higher proliferative response of T cells with concomitant higher expression of interferon-γ, suggesting that the clearance of cell surface sialic acids contributes to the development of a T helper type 1 proinflammatory response. Together, our data strongly support sialic acid’s relevance in DC immune functions. Alterations of cell surface sialic acid content can alter the endocytosis/phagocytosis, maturation, migration/homing and the ability for T cell priming in human DCs. Moreover, sialic acid linkages created by ST6Gal-1 and ST3Gal-1 are functionally relevant. The engineering of cell surface sialylation, mediated by individual sialidases or sialyltransferases is a likely possibility to fine tune DC phagocytosis and immunological potency, with particular significance to DC-based therapies.

Entre a plasticidade do corpo e da imagem : contributos para uma viagem interactiva pelos territórios incertos da arte contemporânea

Este trabalho de investigação começou por ser estruturado em torno de quatro grandes capítulos (quatro grandes linhas de orientação temática), todos eles amplamente desenvolvidos no sentido de podermos cartografar alguns dos principais territórios e sintomas da arte contemporânea, sendo certo também, que cada um deles assenta precisamente nos princípios de uma estrutura maleável que, para todos os efeitos, se encontra em processo de construção (work in progress), neste caso, graças à plasticidade do corpo, do espaço, da imagem e do uso criativo das tecnologias digitais, no âmbito das quais, aliás, tudo se parece produzir, transformar e disseminar hoje em dia à nossa volta (quase como se de uma autêntica viagem interactiva se tratasse). Por isso, a partir daqui, todo o esforço que se segue procurará ensaiar uma hipótese de trabalho (desenvolver uma investigação) que, porventura, nos permita desbravar alguns caminhos em direcção aos intermináveis túneis do futuro, sempre na expectativa de podermos dar forma, função e sentido a um desejo irreprimível de liberdade criativa, pois, a arte contemporânea tem essa extraordinária capacidade de nos transportar para muitos outros lugares do mundo, tão reais e imaginários como a nossa própria vida. Assim sendo, há que sumariar algumas das principais etapas a desenvolver ao longo desta investigação. Ora, num primeiro momento, começaremos por reflectir sobre o conceito alargado de «crise» (a crise da modernidade), para logo de seguida podermos abordar a questão da crise das antigas categorias estéticas, questionando assim, para todos os efeitos, quer o conceito de «belo» (Platão) e de «gosto» (Kant), quer ainda o conceito de «forma» (Foccilon), não só no sentido de tentarmos compreender algumas das principais razões que terão estado na origem do chamado «fim da arte» (Hegel), mas também algumas daquelas que terão conduzido à estetização generalizada da experiência contemporânea e à sua respectiva disseminação pelas mais variadas plataformas digitais. Num segundo momento, procuraremos reflectir sobre alguns dos principais problemas da inquietante história das imagens, nomeadamente para tentarmos perceber como é que todas estas transformações técnicas (ligadas ao aparecimento da fotografia, do cinema, do vídeo, do computador e da internet) terão contribuído para o processo de instauração e respectivo alargamento daquilo que todos nós ficaríamos a conhecer como a nova «era da imagem», ou a imagem na «era da sua própria reprodutibilidade técnica» (Benjamin), pois, só assim é que conseguiremos interrogar este imparável processo de movimentação, fragmentação, disseminação, simulação e interacção das mais variadas «formas de vida» (Nietzsche, Agamben). Entretanto, chegados ao terceiro grande momento, interessa-nos percepcionar a arte contemporânea como uma espécie de plataforma interactiva que, por sua vez, nos levará a interpelar alguns dos principais dispositivos metafóricos e experimentais da viagem, neste caso, da viagem enquanto linha facilitadora de acesso à arte, à cultura e à vida contemporânea em geral, ou seja, todo um processo de reflexão que nos incitará a cartografar alguns dos mais atractivos sintomas provenientes da estética do flâneur (na perspectiva de Rimbaud, Baudelaire, Long e Benjamin) e, consequentemente, a convocar algumas das principais sensações decorrentes da experiência altamente sedutora daqueles que vivem mergulhados na órbita interactiva do ciberespaço (na condição de ciberflâneurs), quase como se o mundo inteiro, agora, fosse tão somente um espaço poético «inteiramente navegável» (Manovich). Por fim, no quarto e último momento, procuraremos fazer uma profunda reflexão sobre a inquietante história do corpo, principalmente com o objectivo de reforçar a ideia de que apesar das suas inúmeras fragilidades biológicas (um ser que adoece e morre), o corpo continua a ser uma das «categorias mais persistentes de toda a cultura ocidental» (Ieda Tucherman), não só porque ele resistiu a todas as transformações que lhe foram impostas historicamente, mas também porque ele se soube reinventar e readaptar pacientemente face a todas essas transformações históricas. Sinal evidente de que a sua plasticidade lhe iria conferir, principalmente a partir do século XX («o século do corpo») um estatuto teórico e performativo verdadeiramente especial. Tão especial, aliás, que basta termos uma noção, mesmo que breve, da sua inquietante história para percebermos imediatamente a extraordinária importância dalgumas das suas mais variadas transformações, atracções, ligações e exibições ao longo das últimas décadas, nomeadamente sob o efeito criativo das tecnologias digitais (no âmbito das quais se processam algumas das mais interessantes operações de dinamização cultural e artística do nosso tempo). Em suma, esperamos sinceramente que este trabalho de investigação possa vir a contribuir para o processo de alargamento das fronteiras cada vez mais incertas, dinâmicas e interactivas do conhecimento daquilo que parece constituir, hoje em dia, o jogo fundamental da nossa contemporaneidade.

Peer interaction and learning opportunities in cohesive and less cohesive L2 classrooms

The present study investigates peer to peer oral interaction in two task based language teaching classrooms, one of which was a self-declared cohesive group, and the other a self- declared less cohesive group, both at B1 level. It studies how learners talk cohesion into being and considers how this talk leads to learning opportunities in these groups. The study was classroom-based and was carried out over the period of an academic year. Research was conducted in the classrooms and the tasks were part of regular class work. The research was framed within a sociocognitive perspective of second language learning and data came from a number of sources, namely questionnaires, interviews and audio recorded talk of dyads, triads and groups of four students completing a total of eight oral tasks. These audio recordings were transcribed and analysed qualitatively for interactions which encouraged a positive social dimension and behaviours which led to learning opportunities, using conversation analysis. In addition, recordings were analysed quantitatively for learning opportunities and quantity and quality of language produced. Results show that learners in both classes exhibited multiple behaviours in interaction which could promote a positive social dimension, although behaviours which could discourage positive affect amongst group members were also found. Analysis of interactions also revealed the many ways in which learners in both the cohesive and less cohesive class created learning opportunities. Further qualitative analysis of these interactions showed that a number of factors including how learners approach a task, the decisions they make at zones of interactional transition and the affective relationship between participants influence the amount of learning opportunities created, as well as the quality and quantity of language produced. The main conclusion of the study is that it is not the cohesive nature of the group as a whole but the nature of the relationship between the individual members of the small group completing the task which influences the effectiveness of oral interaction for learning.This study contributes to our understanding of the way in which learners individualise the learning space and highlights the situated nature of language learning. It shows how individuals interact with each other and the task, and how talk in interaction changes moment-by-moment as learners react to the ‘here and now’ of the classroom environment.

DISTRIBUTION AND FEEDING ECOLOGY OF THE AFRICAN TILAPIA Oreochromis mossambicus (TELEOSTEI, PERCIFORMES, CICHLIDAE) IN SURINAME (SOUTH AMERICA) WITH COMMENTS ON THE TILAPIA-KWIKWI (Hoplosternum littorale) (TELEOSTEI, SILURIFORMEs, CALLICHTHYIDAE) INTERACTION

The geographical distribution of the African Tilapia Oreochromis mossambicusin Suriname is restricted to a narrow strip of land along the Atlantic coast. Within the coastal plain, O. mossambicusoccurs in brackish lagoons, oligohaline canals, and shell-sand pit lakes. Physico-chemical characteristics and phytoplankton composition of representative Tilapia water bodies are described. Blue-green algae and fine flocculent detritus are dominant food items in the diet of the Tilapia, while Rotifera and microcrustacea are also important in the diet of larvae and juveniles. Intraspecific diet overlap among ontogenetic stages of the Tilapia did not differ significantly from 1, which means that these diets showed complete overlap. Interspecific diet overlap between the Tilapia and the indigenous armoured catfish Hoplosternum littoralewere moderate or low. The results are discussed in relation to recent developments in the Surinamese fisheries and aquaculture sector.

Xenogeneic M2-polarization of Macrophages by undifferentiated and differentiated adipose tissue-derived stem cells

Mesenchymal stem cells (MSCs) are considered to be â â immunologically privileged.â â In a previous work when human adipose tissue-derived stem cells (hASCs) subcutaneously implanted in mice we did not identify an adverse host response1. Recently, it was shown that tissue regeneration could benefit from the polarization of M2 macrophages subpopulations 2. In this study we hypothesised that undifferentiated hASCs and derived osteoblasts and chondrocytes are able to switch murine bone marrow-derived macrophages (mBMMÃ s) into M2 phenotype, aiding tissue regeneration. Murine BMMÃ s were plated in direct contact with undifferentiated and osteo or chondro-differentiated hASCs for 4 h, 10 h, 24 h and 72 h. The cytokine profile was analysed by qRT-PCR and the surface markers were detected by flow cytometry. The direct interaction of both cell types was observed by time lapse microscopy. The results showed that mBMMÃ s polarized after contacting tissue culture polystyrene. This M2 phenotype was maintained along the experiment in direct contact with both undifferentiated and osteo or chondro-differentiated hASCs. This was confirmed by the expression of IL-1, IL-10, IL-4, TNF-a and IFN-g (genetic profile) and surface markers (CD206 + + , CD336 + + , MHC II + and CD86 + + ) detection. These data suggest the potential of hASCs in contemporary xenogenic tissue engineering and regenerative medicine strategies, as well as host immune system modulation in autoimmune diseases. 

The impact of PBL on transferable skills development in management education

Attention to transferable skills is growing in higher education. Problem-based learning (PBL) is increasingly used in management education for its promising potential to, amongst others, promote transferable skills, including problem-solving, critical thinking and teamwork skills. However, this relationship is seldom assessed. In this study, I use structural equation modelling to examine the effectiveness of PBL, measured through students’ perceptions of satisfaction and skills development. Results show the development of transferable skills is explained by interaction with tutors and a host company, and defining teamwork rules. Satisfaction is explained by skills development, assessment issues, defining teamwork rules and understanding how organisations work. I draw conclusions and recommendations.

Study of the interaction between asphalt and plastic wastes in new polymer modified binders (PMB)

The aim of this study is evaluating the interaction between several base pen grade asphalt binders (35/50, 50/70, 70/100, 160/220) and two different plastic wastes (EVA and HDPE), for a set of new polymer modified binders produced with different amounts of both plastic wastes. After analysing the results obtained for the several polymer modified binders evaluated in this study, including a commercial modified binder, it can be concluded that the new PMBs produced with the base bitumen 70/100 and 5% of each plastic waste (HDPE or EVA) results in binders with very good performance, similar to that of the commercial modified binder.

Hand gesture recognition for human computer interaction : a comparative study of different image features

Hand gesture recognition for human computer interaction, being a natural way of human computer interaction, is an area of active research in computer vision and machine learning. This is an area with many different possible applications, giving users a simpler and more natural way to communicate with robots/systems interfaces, without the need for extra devices. So, the primary goal of gesture recognition research is to create systems, which can identify specific human gestures and use them to convey information or for device control. For that, vision-based hand gesture interfaces require fast and extremely robust hand detection, and gesture recognition in real time. In this study we try to identify hand features that, isolated, respond better in various situations in human-computer interaction. The extracted features are used to train a set of classifiers with the help of RapidMiner in order to find the best learner. A dataset with our own gesture vocabulary consisted of 10 gestures, recorded from 20 users was created for later processing. Experimental results show that the radial signature and the centroid distance are the features that when used separately obtain better results, with an accuracy of 91% and 90,1% respectively obtained with a Neural Network classifier. These to methods have also the advantage of being simple in terms of computational complexity, which make them good candidates for real-time hand gesture recognition.

Generic system for human-computer gesture interaction: applications on sign language recognition and robotic soccer refereeing

Hand gestures are a powerful way for human communication, with lots of potential applications in the area of human computer interaction. Vision-based hand gesture recognition techniques have many proven advantages compared with traditional devices, giving users a simpler and more natural way to communicate with electronic devices. This work proposes a generic system architecture based in computer vision and machine learning, able to be used with any interface for human-computer interaction. The proposed solution is mainly composed of three modules: a pre-processing and hand segmentation module, a static gesture interface module and a dynamic gesture interface module. The experiments showed that the core of visionbased interaction systems could be the same for all applications and thus facilitate the implementation. For hand posture recognition, a SVM (Support Vector Machine) model was trained and used, able to achieve a final accuracy of 99.4%. For dynamic gestures, an HMM (Hidden Markov Model) model was trained for each gesture that the system could recognize with a final average accuracy of 93.7%. The proposed solution as the advantage of being generic enough with the trained models able to work in real-time, allowing its application in a wide range of human-machine applications. To validate the proposed framework two applications were implemented. The first one is a real-time system able to interpret the Portuguese Sign Language. The second one is an online system able to help a robotic soccer game referee judge a game in real time.

Generic system for human-computer gesture interaction

Hand gestures are a powerful way for human communication, with lots of potential applications in the area of human computer interaction. Vision-based hand gesture recognition techniques have many proven advantages compared with traditional devices, giving users a simpler and more natural way to communicate with electronic devices. This work proposes a generic system architecture based in computer vision and machine learning, able to be used with any interface for humancomputer interaction. The proposed solution is mainly composed of three modules: a pre-processing and hand segmentation module, a static gesture interface module and a dynamic gesture interface module. The experiments showed that the core of vision-based interaction systems can be the same for all applications and thus facilitate the implementation. In order to test the proposed solutions, three prototypes were implemented. For hand posture recognition, a SVM model was trained and used, able to achieve a final accuracy of 99.4%. For dynamic gestures, an HMM model was trained for each gesture that the system could recognize with a final average accuracy of 93.7%. The proposed solution as the advantage of being generic enough with the trained models able to work in real-time, allowing its application in a wide range of human-machine applications.

Interaction paradigms versus age-related user profiles: an evaluation on content selection

Novel input modalities such as touch, tangibles or gestures try to exploit human's innate skills rather than imposing new learning processes. However, despite the recent boom of different natural interaction paradigms, it hasn't been systematically evaluated how these interfaces influence a user's performance or whether each interface could be more or less appropriate when it comes to: 1) different age groups; and 2) different basic operations, as data selection, insertion or manipulation. This work presents the first step of an exploratory evaluation about whether or not the users' performance is indeed influenced by the different interfaces. The key point is to understand how different interaction paradigms affect specific target-audiences (children, adults and older adults) when dealing with a selection task. 60 participants took part in this study to assess how different interfaces may influence the interaction of specific groups of users with regard to their age. Four input modalities were used to perform a selection task and the methodology was based on usability testing (speed, accuracy and user preference). The study suggests a statistically significant difference between mean selection times for each group of users, and also raises new issues regarding the “old” mouse input versus the “new” input modalities.

(Un)suitability of the use of pH buffers in biological, biochemical and environmental studies and their interaction with metal ions - a review

The use of buffers to maintain the pH within a desired range is a very common practice in chemical, biochemical and biological studies. Among them, zwitterionic N-substituted aminosulfonic acids, usually known as Good's buffers, although widely used, can complex metals and interact with biological systems. The present work reviews, discusses and updates the metal complexation characteristics of thirty one commercially available buffers. In addition, their impact on biological systems is also presented. The influences of these buffers on the results obtained in biological, biochemical and environmental studies, with special focus on their interaction with metal ions, are highlighted and critically reviewed. Using chemical speciation simulations, based on the current knowledge of the metal-buffer stability constants, a proposal of the most adequate buffer to employ for a given metal ion is presented.

Estudo de validação do Partner Interaction Questionnaire para fumantes e ex-fumantes

O presente artigo tem por objetivo validar uma versão adaptada do Partner Interaction Questionnaire (PIQ), instrumento que avalia o suporte para deixar de fumar dado pelo parceiro. É apresentada uma versão adaptada do PIQ para fumantes e abstinentes. Participaram no estudo 224 fumantes (52.7% do sexo feminino e 47.3% do sexo masculino; média de idades=28.6 anos; DP=8.69) e 169 ex-fumantes (68.6% do sexo masculino e 31.4% do sexo feminino; média de idades= 43.1 anos; DP=13.02). O instrumento apresentou boa fidelidade (alfa de Cronbach: 0.73) para a subescala positiva e para a subescala negativa (alfa de Cronbach: 0.77). O estudo de validade revelou a presença de dois fatores que explicam 43.36% da variância. O instrumento apresenta boa validade de construto, correlacionando-se positivamente com o ajustamento de casais e a qualidade de vida mental, e negativamente com a depressão. Esta versão adaptada do PIQ reuniu as condições necessárias para a sua aplicação em fumantes e ex-fumantes, podendo ser uma escala útil em programas de intervenção de cessação tabágica e de manutenção da abstinência.

Thin films composed of gold nanoparticles dispersed in a dielectric matrix: the influence of the host matrix on the optical and mechanical responses

Gold nanoparticles were dispersed in two different dielectric matrices, TiO2 and Al2O3, using magnetron sputtering and a post-deposition annealing treatment. The main goal of the present work was to study how the two different host dielectric matrices, and the resulting microstructure evolution (including both the nanoparticles and the host matrix itself) promoted by thermal annealing, influenced the physical properties of the films. In particular, the structure and morphology of the nanocomposites were correlated with the optical response of the thin films, namely their localized surface plasmon resonance (LSPR) characteristics. Furthermore, and in order to scan the future application of the two thin film system in different types of sensors (namely biological ones), their functional behaviour (hardness and Young's modulus change) was also evaluated. Despite the similar Au concentrations in both matrices (~ 11 at.%), very different microstructural features were observed, which were found to depend strongly on the annealing temperature. The main structural differences included: (i) the early crystallization of the TiO2 host matrix, while the Al2O3 one remained amorphous up to 800 °C; (ii) different grain size evolution behaviours with the annealing temperature, namely an almost linear increase for the Au:TiO2 system (from 3 to 11 nm), and the approximately constant values observed in the Au:Al2O3 system (4–5 nm). The results from the nanoparticle size distributions were also found to be quite sensitive to the surrounding matrix, suggesting different mechanisms for the nanoparticle growth (particle migration and coalescence dominating in TiO2 and Ostwald ripening in Al2O3). These different clustering behaviours induced different transmittance-LSPR responses and a good mechanical stability, which opens the possibility for future use of these nanocomposite thin film systems in some envisaged applications (e.g. LSPR-biosensors).

Paving the way for predictive diagnostics and personalized treatment of invasive aspergillosis

Invasive aspergillosis (IA) is a life-threatening fungal disease commonly diagnosed among individuals with immunological deficits, namely hematological patients undergoing chemotherapy or allogeneic hematopoietic stem cell transplantation. Vaccines are not available, and despite the improved diagnosis and antifungal therapy, the treatment of IA is associated with a poor outcome. Importantly, the risk of infection and its clinical outcome vary significantly even among patients with similar predisposing clinical factors and microbiological exposure. Recent insights into antifungal immunity have further highlighted the complexity of host-fungus interactions and the multiple pathogen-sensing systems activated to control infection. How to decode this information into clinical practice remains however, a challenging issue in medical mycology. Here, we address recent advances in our understanding of the host-fungus interaction and discuss the application of this knowledge in potential strategies with the aim of moving toward personalized diagnostics and treatment (theranostics) in immunocompromised patients. Ultimately, the integration of individual traits into a clinically applicable process to predict the risk and progression of disease, and the efficacy of antifungal prophylaxis and therapy, holds the promise of a pioneering innovation benefiting patients at risk of IA.