Identificação de novas espécies de Brucella spp.: novas ameaças para os humanos

Tese de mestrado, Microbiologia Aplicada, Universidade de Lisboa, Faculdade de Ciências, 2022A brucelose á uma doença zoonótica mundial causada por Brucella spp., uma alfaproteobactéria Gram-negativa, capaz de infetar uma vasta gama de mamíferos, que incluem animais domésticos, animais selvagens e seres humanos. Dentro do gênero de Brucella estão descritas 12 espécies, três das quais (Brucella abortus, Brucella suis e Brucella melitensis) estão mais frequentemente associadas à brucelose humana, comumente designadas por espécies clássicas. Desde o final dos anos 90, foram isoladas a partir de seres humanos, animais selvagens e fontes ambientais várias espécies de Brucella spp., evidenciando a existência de uma gama de hospedeiros mais ampla. Algumas destas espécies recentemente descritas são geneticamente e/ou fenotipicamente diferentes quando comparadas com as espécies clássicas, o que pode resultar em potenciais novas ameaças zoonóticas. Nos dias de hoje, apenas algumas espécies clássicas de Brucella spp. estão sujeitas a estratégias de controlo e vigilância na Europa, enquanto as novas espécies detetadas se encontram potencialmente subdiagnosticadas na vida selvagem. Com o presente trabalho, e através de uma abordagem One Health, pretendeu-se realizar a identificação e caraterização de novas espécies de Brucella spp. através de soros humanos, pertencentes à seroteca do Instituto Nacional de Saúde Doutro Ricardo Jorge, tanto com recurso a metodologias de diagnóstico diretas, como o teste de aglutinação Rosa de Bengala, tal como através de técnicas moleculares, como PCR em tempo real. De um total de 168 amostras analisadas, apenas 21 apresentaram um resultado positivo através de PCR e, após sequenciação, nenhuma das amostras apresentou o genoma correspondente a Brucella spp. Estes resultados realçam as dificuldades inerentes ao diagnóstico deste microrganismo, podendo as mesmas ajudar na otimização da caraterização e tipagem de Brucella em trabalhos futuros. Igualmente neste trabalho, e utilizando uma abordagem inovadora, recorrendo às novas metodologias de sequenciação de nova geração, foi possível validar a utilização do perfil de MLVA-16, a técnica gold standard para a tipagem de Brucella spp., utilizando a extração de genótipos in silico. Neste sentido, pretendia-se avaliar o desempenho de um script Python, comparando-o com a abordagem de MLVA baseada em PCR, em 83 estirpes de diferentes espécies de Brucella. A abordagem de MLVA baseada em WGS detetou 95,3% de todos os 1 328 possíveis hits (83 estirpes x 16 loci) e mostrou uma taxa de concordância de 96,4% com o procedimento de MLVA clássico baseado em PCR. Este script parece ser uma ferramenta muito útil e robusta para a determinação in silico dos perfis de MLVA para as estirpes de Brucella, permitindo estudos epidemiológicos moleculares, importantes para a manutenção de uma vigilância ativa da brucelose

Relationship between physical activity levels of Portuguese physical therapists and mental health during a COVID-19 pandemic: being active is the key

FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.Introduction: Physical activity is essential for a healthy life and quality of life, representing a fundamental role in individuals' physical and mental health. Concomitantly, the physical therapist, through the promotion of physical activity and exercise, can improve mental health, an essential factor in the current pandemic, triggering anxiety, fear, and depression crisis. Objective: To verify physical activity among Portuguese physical therapists and its association with mental health during the pandemic times. Methods: An online questionnaire was applied through social media to all Portuguese physical therapists between October 21, 2021, and January 14, 2022. It contained general characterization questions of the sample, the IPAQ-SF questionnaire to assess physical activity levels, the Goldberg General Health Questionnaire (GHQ-28), which assesses the levels of mental health, and the WHO Well-Being Index (WHO-5) to assess subjective well-being. Results: The sample totaled 286 respondents (82% female), with a mean age of 33. Of the total answers, 82% practiced physical activity, 45% had moderate levels of physical activity, and 19% were vigorous. Physical therapists in these categories had lower values in the GHQ-28 and higher in the WHO-5. Those with better mental health also showed better subjective well-being (r = -0.571, p = 0.000). Conclusion: The data obtained showed that physical therapists mostly have moderate and vigorous levels of physical activity and that physical activity positively influences individuals' mental health and well-being, which proved to be a key factor due to the pandemic situation.info:eu-repo/semantics/publishedVersio

O Parque Florestal de Monsanto. Evolução histórica e contributo para a sua gestão

Mestrado em Arquitetura Paisagista - Instituto Superior de AgronomiaMonsanto Forest Park is the largest park in Lisbon, stretching over 1000 ha of land. Historical records dating back as far as 1868 clearly display an intention of creating a forest park over what was then the bare mountain of Monsanto. However, these plans did not turn into reality until the 1930’s, when the Minister for Public Works Eng. Duarte Pacheco, helped by the acute hand of Arch. Keil do Amaral. At the dawn of the 21st century, the city of Lisbon registers an average of green spaces of 9,1 m2/inhab (without Monsanto Park), and of 26,8 m2/inhab if we include it (Soares and Castel-Branco, 2007). Such numbers state and justify the importance of the theme of this work, which is not only a biophysical and historical analysis of the Park, but also aims to portrait its evolution. The final goal of this work is to contribute with relevant considerations for the Park’s management, taking into account the Forest Management Plan approved by the National Forest Authority in 2012 and a survey made to the park’s users, to outline guidelines which aim for possibly turning the plan into a real-life park managing experience, attending to its ecological, aesthetic and social components

Communication in health care and patient safety: problems and challenges

A segurança do doente constitui um dos grandes desafios dos cuidados de saúde do séc. XXI. O reconhecimento da ocorrência de erros ou acidentes adversos com consequências gravosas para os doentes e para as instituições de saúde, levou, recentemente, a Organização Mundial de Saúde (OMS) a nomear comissões centradas na identificação de situações de risco e na elaboração de soluções que possam servir de recurso para a prevenção dessas situações. O resultado dos trabalhos destas comissões tornou evidente a importância da comunicação como determinante da qualidade e da segurança na prestação de cuidados. Neste artigo é abordada a problemática da comunicação em saúde e a sua relação com a segurança do doente, identificados problemas e apontadas algumas pistas para a sua prevenção. São especificamente referidos os principais problemas de comunicação entre os profissionais de saúde (nas passagens de turno e nas equipas de saúde) e entre estes profissionais e os doentes.ABSTRACT - Patient safety is a major challenge for the XXI century health care. The recognition of inadvertent harm to patients as result of professional health care has, recently, led the World Health Organization (WHO) to appoint committees focused on identifying risk situations and developing solutions that can serve as a resource for the prevention of these adverse events. The outcome of these commissions has highlighted the importance of communication as a determinant of quality and safety in health care. In this article we address the issue of communication in health care and its relationship to patient safety, identifying problems and pointing out some clues to its prevention

Estudo dos efeitos do metilfenidato em medidas de funções executivas e comportamentais em rapazes com PHDA-M : análise em dois tempos

Tese de mestrado, Psicologia (Secção de Psicologia Clínica e da Saúde - Núcleo de Psicologia da Saúde e da Doença), Universidade de Lisboa, Faculdade de Psicologia, 2013A Perturbação de Hiperatividade com Défice de Atenção (PHDA) é considerada a perturbação psiquiátrica do desenvolvimento mais diagnosticada na infância. A dificuldade no diagnóstico e terapêutica de esta Perturbação decorre da discussão que a rodeia e que tem origem na controvérsia existente na definição dos seus determinantes causais, bem como do seu diagnóstico e terapêutica. Neste estudo pretende-se avaliar o efeito da terapêutica psico-estimulante (metilfenidato) nas funções executivas e nos comportamentos externalizantes e internalizantes, numa amostra de rapazes diagnosticados com PHDA tipo misto. Trata-se de um estudo onde os participantes são sujeitos a uma avaliação psicológica em dois tempos, com e sem o efeito de terapêutica psico-estimulante. Os principais resultados apontam para uma evolução positiva em apenas algumas medidas de funções executivas como a velocidade de processamento e a memória de trabalho verbal, mas não demonstra ajudar estas crianças e adolescentes a aumentarem a sua qualidade de vida e a sua realização pessoal. Parece clara a necessidade de uma avaliação neuropsicológica e emocional como suporte à definição do diagnóstico, assim como de uma intervenção que não se baseie apenas na medicação por psico-estimulantes.Attention-Deficit Hyperactivity Disorder (ADHD) is the most diagnosed developmental psychiatric disorder in children. The difficulties in the diagnosis and treatment of ADHD are a direct consequence of the existing controversy surrouding the definitions of its causal determinants, as well as its diagnosis criterions and adequate treatment measures. In this study it will be examined the effects of a psico-stimulant medication (methylphenidate) in executive functions and externalizing and internalizing behaviours, using a sample of boys with ADHD Combinaned Type (ADHD-C). In this study, the boys will be subjected to a psychological assessment in two different moments; in which they are evaluated with and without psico-stimulant medication. The main results indicate a positive comparison between the two moments in some of the measures of executive functions, such as processing speed and verbal working memory, but this psico-stimulant doesn't seems to help this children and adolescents enhancing their quality of life , personal well-being or self-fulfilment. The urge to an neuropsychological and emotional evaluation as a part of the diagnosis criterions seems very clear, and the same urgency is found in the need of treatments not relying exclusively on psicostimulants medication

Whole Genome Sequencing-Based Approach for the Determination of Multiple Locus Variable Number Tandem Repeat Profiles

Funding Information: Funding: This work is a result of the GenomePT project (POCI-01- 0145-FEDER-022184), supported by the COMPETE 2020 – Operational Program for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Program (Lisboa2020), Algarve Portugal Regional Operational Program (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by the Fundação para a Ciência e a Tecnologia (FCT). The studies have arisen from the Project QUANDHIP (Chafea Grant Agreement no. 2010 21 02), which has been funded by the European Commission in the framework of the Health Program.Brucellosis is an important zoonosis that is emerging in some regions of the world, gaining increased relevance with the inclusion of the causing agent Brucella spp. in the class B bioterrorism group. Until now, multi-locus VNTR Analysis (MLVA) based on 16 loci has been considered as the gold standard for Brucella typing. However, this methodology is laborious, and, with the rampant release of Brucella genomes, the transition from the traditional MLVA to whole genome sequencing (WGS)-based typing is on course. Nevertheless, in order to avoid a disruptive transition with the loss of massive genetic data obtained throughout the last decade and considering that the transition timings will vary considerably among different countries, it is important to determine WGS-based MLVA alleles of the nowadays sequenced genomes. On this regard, we aimed to evaluate the performance of a Python script that had been previously developed for the rapid in silico extraction of the MLVA alleles, by comparing it to the PCR-based MLVA procedure over 83 strains from different Brucella species. The WGS-based MLVA approach detected 95.3% of all possible 1,328 hits (83 strains×16 loci) and showed an agreement rate with the PCR-based MLVA procedure of 96.4% for MLVA-16. According to our dataset, we suggest the use of a minimal depth of coverage of ~50x and a maximum number of ~200 contigs as guiding “boundaries” for the future application of the script. In conclusion, the evaluated script seems to be a very useful and robust tool for the in silico determination of MLVA profiles of Brucella strains, allowing retrospective and prospective molecular epidemiological studies, which are important for maintaining an active epidemiological surveillance of brucellosis.publishersversionpublishe

Role of MurT C-Terminal Domain in the Amidation of Staphylococcus aureus Peptidoglycan

Fundacao para a Ciencia e a Tecnologia (FCT) through grants PTDC/FIS-NAN/ 0117/2014 and PTDC/BIA-MIC/31645/2017. project UID/Multi/04378/2019 (Unidade de Ciencias Biomoleculares Aplicadas-UCIBIO), funded by FCT/MCTES; project LISBOA-01-0145-FEDER-007660 (Microbiologia Molecular, Estrutural e Celular), funded by FEDER through COMPETE2020-Programa Operacional Competitividade e Internacionalizacao (POCI); national funds through FCT; by project ONEIDA (LISBOA-01-0145-FEDER-016417), cofunded by FEEI (Fundos Europeus Estruturais e de Investimento) from the Programa Operacional Regional Lisboa 2020; and by national funds from FCT. Funding was also provided by European Society of Clinical Microbiology and Infectious Diseases research grant 2015, awarded to R.G.S. B.V.G., T.A.F., and I.R.G. were supported by fellowships SFRH/BD/131623/2017 respectively. J.S.D. acknowledges the National NMR Network (PTNMR) and Infrastructure Project ROTEIRO/0031/2013-PINFRA/22161/2016 (cofinanced by FEDER through COMPETE 2020, POCI, PORL, and FCT through PIDDAC).Glutamate amidation, a secondary modification of the peptidoglycan, was first identified in Staphylococcus aureus. It is catalyzed by the protein products of the murT and gatD genes, which are conserved and colocalized in the genomes of most sequenced Gram-positive bacterial species. The MurT-GatD complex is required for cell viability, full resistance to β-lactam antibiotics, and resistance to human lysozyme and is recognized as an attractive target for new antimicrobials. Great effort has been invested in the study of this step, culminating recently in three independent reports addressing the structural elucidation of the MurT-GatD complex. In this work, we demonstrate through the use of nonstructural approaches the critical and multiple roles of the C-terminal domain of MurT, annotated as DUF1727, in the MurT-GatD enzymatic complex. This domain provides the physical link between the two enzymatic activities and is essential for the amidation reaction. Copurification of recombinant MurT and GatD proteins and bacterial two-hybrid assays support the observation that the MurT-GatD interaction occurs through this domain. Most importantly, we provide in vivo evidence of the effect of substitutions at specific residues in DUF1727 on cell wall peptidoglycan amidation and on the phenotypes of oxacillin resistance and bacterial growth.publishersversionpublishe

VII Workshop on Computational Data Analysis and Numerical Methods

Dear participants, colleagues and friends, it is a great honour and a privilege to give you all a warmest welcome to the VII Workshop on Computational Data Analysis and Numerical Methods (VII WCDANM), which is organized by the Polytechnic Institute of Tomar (located in the center of Portugal in the beautiful city of Tomar), with the support of some Portuguese research centers, hoping that the final result may exceed the expectations of the participants, sponsors and organizers. Due to the worldwide pandemic caused by the COVID-19 virus, for the first time, this meeting will be transmitted through videoconference (webminar). Nevertheless, the important contributions of Adélia Sequeira (University of Lisbon, Portugal), Sílvia Barbeiro (University of Coimbra, Portugal), Malay Banerjee (Indian Instituto of Techonology Kampur, India) and Indranhil Ghosh (University of North Carolina at Wilmington, USA) as Plenary Speakers, the high scientific level of oral and poster presentations and an active audience will certainly contribute to the success of the meeting. Part of the accepted papers (theoretical and applied) by the VII WCDANM involve big data, data mining, data science and machine learning, in different areas of research, some giving emphasis to coronavirus. A very special thanks to this small, yet important, scientific community, since this event could not be possible without any of these essential and complementary parts. This year, there is also the possibility to attend a course on ŞModeling Partial Least Squares Structural Equations (PLS-SEM) using SmartPLSŤ given by Christian M. Ringle ((TUHH) Hamburg University of Technology, Germany), who kindly and readily accepted our invitation and to whom we are very grateful. A special acknowledgment is also due to the Members of the Executive, Scientific and Organizing Committees. In particular, Anuj Mubayi (Arizona StateUniversity,USA),MilanStehlík(Johannes KeplerUniversity,Austria), AnaNata,IsabelPitacasandManuelaFernandes(hostsfromthePolytechnic Institute of Tomar, Portugal), A. Manuela Gonçalves (University of Minho, Portugal), Teresa Oliveira (Aberta University) and Fernando Carapau (University of Évora, Portugal) have been relentless in search for a balanced, broad and interesting program, having achieved an excellent result. For the third consecutive year, the Journal of Applied Statistics (Taylor & Francis) and Neural Computing and Applications (Springer) are also associated to the event, being extremely important in the dissemination of the scientific results achieved at the meeting. Given the above, it is a pleasure to be ”together” with all of you in this web conference, hoping it may provide an intellectual stimulus and an opportunity for the scientific community to jointly work and disseminate scientific research, namely presenting approaches that may contribute to the solution of the pandemic we are experiencing in the expectation that the present might be past in the near future

Estimation of the allergenic potential of urban trees and urban parks: Towards the healthy design of urban green spaces of the future

The impact of allergens emitted by urban green spaces on health is one of the main disservices of ecosystems. The objective of this work is to establish the potential allergenic value of some tree species in urban environments, so that the allergenicity of green spaces can be estimated through application of the Index of Urban Green Zones Allergenicity (IUGZA). Multiple types of green spaces in Mediterranean cities were selected for the estimation of IUGZ. The results show that some of the ornamental species native to the Mediterranean are among the main causative agents of allergy in the population; in particular, Oleaceae, Cupressaceae, Fagaceae, and Platanus hispanica. Variables of the strongest impact on IUGZA were the bioclimatic characteristics of the territory and design aspects, such as the density of trees and the number of species. We concluded that the methodology to assess the allergenicity associated with urban trees and urban areas presented in this work opens new perspectives in the design and planning of urban green spaces, pointing out the need to consider the potential allergenicity of a species when selecting plant material to be used in cities. Only then can urban green areas be inclusive spaces, in terms of public health.This work has been made possible thanks to members of the Silva MediterraneaWorking Group on Urban and Peri-Urban Forestry (FAO WG7) and COST Action FP1204 Green Infrastructure approach: linking environmental with social aspects in studying and managing urban forests. Nezha Acil thanks the technical sta of ISESCO park for their help during his first visit to the park. Maria Beatrice Andreucci also thanks students Giada Di Sante, Guglielmo Pirri e Daniele Purini for her course in Environmental Technological Design of the Master degree in Landscape Architecture at Sapienza Università di Roma for participation in the tree inventory and Duilio Iamonico for the tree taxonomy classification review. The research by the Slovenian Forestry Institute was financially supported by the Program and Research group P4-0107 “Forest ecology, biology and technology” funded by the Slovenian Research Agency.info:eu-repo/semantics/publishedVersio