Novel nanoparticle-based therapy to eradicate Pseudomonas aeruginosa infections in cystic fibrosis lungs: dual drug release by cubosomes

Book of Abstracts of CEB Annual Meeting 2017info:eu-repo/semantics/publishedVersio

Small colony variants in biofilm population is not strictly required for biofilm resistance and persistence

Pseudomonas aeruginosa is a well-known opportunistic pathogen, responsible for high mortality in several human infections. Such infection success is partly due to its phenotypic plasticity to switch to “fitter” phenotypes, such as small colony variants (SCV), generally associated to increased antibiotic resistance, and biofilm formation within the human body. Once embedded in biofilms, P. aeruginosa can augment its pathogenicity and improve its ability to survive to stressful conditions, such immune defenses and antibiotics action. Moreover, the presence of SCV in biofilms has been pointed out as a biofilm mechanism responsible for its antibiotic resistance and persistence in human body. Regarding the impact of P. aeruginosa biofilms in disease management, it was considered crucial to determine the role of SCV in biofilm population and to study its correlation with antibiotic resistance and biofilm persistence ability. This study used several P. aeruginosa strains to form distinct biofilms in order to assess their antibiotic susceptibility to ciprofloxacin and colistin, virulence factors expression of biofilm-associated bacteria and to determine the presence and prevalence of SCV in biofilm population. The results obtained revealed that SCV presence in biofilm population is not strictly associated with biofilm antibiotic resistance. P. aeruginosa PAI2 demonstrated augmented resistance to both antibiotics, as other strains, but without the presence of SCV in biofilm population. Persistence was as well not strictly associated with SCV since CECT 111 biofilm-associated bacteria did not produced SCV but expressed relevant virulence factors (hemolysin, slime and piocyanin production, and motility) crucial to persistence in human body

Pedelec on a hilly city: a case study in Coimbra

Conclusions: The pedelec emerges from this research as a viable active transport alternative for hilly cities. With adequate autonomy and power to overcome relief, one of the greatest deterrents for using the bicycle as main transport mode, it gives the ball back to politicians, who are responsible for providing cyclists with safe and comfortable pathways for exercising their choice for this mode. Investment on bike paths, bike lanes and adjustment of crossings is the natural next step to foster a more active and sustainable transport mode, from which all citizens would benefit.The pedelec is a basic type of electrically-assisted bicycle, designed to help cyclists overcome most slopes. This article presents a theoretical case study of a pedelec on the city of Coimbra, Portugal, which shows that even a low-end model provides enough power and autonomy for the daily transportation needs of a regular cyclist. It is only if battery wear is considered that a model of higher battery capacity is recommended, and even then only if the cyclist uses the pedelec very intensively. By overcoming what is arguably one of the greatest deterrents to cycling, the pedelec contributes towards materializing the concept of sustainable mobility. Overcoming the other major deterrent, safety concerns, is thus passed to the political decision makers, which will need to build and improve the urban infrastructures necessary for citizens to safely exercise their choice for this transport mode.info:eu-repo/semantics/publishedVersio

Indirect effect of 7-valent and 13-valent pneumococcal conjugated vaccines on pneumococcal pneumonia hospitalizations in elderly

Background: Pneumonia is one of the leading causes of mortality and has a high burden in morbidity. In Portugal, 7-valent pneumococcal conjugated vaccine (PCV) was used since 2001 and PCV10/13 since 2009, being the last introduced into the National Immunization Program in 2015. Methods: We conducted an ecological study to evaluate the impact of PCV7 and PCV13 on pneumococcal pneumonia (PP) hospitalizations in adults aged 65 years or more in Portugal. National hospital discharge registry data from 1998/99 to 2015/16 were used, and PP hospitalization was defined as any hospitalization coded in primary diagnosis as 481 (ICD-9-CM) or J18 (ICD-10-CM). Poisson regression models adjusted for seasonality, influenza-like illness and allowing for overdispersion was used to estimate annual average change of PP hospitalization rate. To assess PP hospitalization trends before and after PCV7 and PCV13 introduction interrupted time series analysis was performed. Results: In 1998/99 PP hospitalization rate was 7.0 per 10,000 inhabitants, varying between 3.2 (females, 65-74 years) to 20.7 (males, +85 years), and annually increasing by 16% during the pre-PCV7 period. Statistically significant reduction of 14% per year in PP hospitalization rate was observed after PCV7 introduction. Between 2004/05 and 2009/10 PP hospitalization rate decreased annually by 4% and after PCV13 introduction by 11% per year. In 2015/16 we found an overall reduction of 2.9 (CI 95%: 2.7; 3.1) PP hospitalizations per 10,000 inhabitants (598 hospitalizations) attributable to PCV13, varying from 2.2 (CI 95%: 1.3; 3.1) (female, 65-74 years) to 5.6 (CI 95%: 3.8; 7.5) (female, +85 years). Conclusions: Our results suggest that introduction of both PCV7 and PCV13 vaccines resulted in the reduction of PP hospitalizations rates among older adults.The IMOVE+ project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 634446info:eu-repo/semantics/publishedVersio

Insights into bacterial colony morphology evolution and diversification during infection development in cystic fibrosis

Tese de Doutoramento em Engenharia Biomédica.Pseudomonas aeruginosa infections are the major cause of high morbidity and mortaiity in cystic fibrosis (CF) patients. Despite the long and aggressive antibiotic treatments, P. aeruginosa still persists causing chronic infections. Its Iong-persistence is due to sophisticated mechanisms of adaptation, including cional diversification into specialized CF-adapted phenotypes. While CF community awaits the development of effective therapies targeting conductance regulator mutations, the gold standard of CF disease management is the eradication of P. aeruginosa from CF lungs as soon as detected. Early eradication of P. aeruginosa avoids or, at ieast, retards the development of chronic infections preserving lung function. But to achieve a successful eradication, it is need understanding the eariy adaptations mechanisms used by P. aeruginosa for antimicrobial agents target them. Up to now these mechanisms are unclear. Being so, this project aimed to determine the early adaptations undergone by P. aeruginosa after CF lungs colonization and their driven forces in several clinical scenarios. Alterations in colony morphology are one of the indicators of cional diversification of bacteria in CF iungs and are also associated with different virulence factors expression and antibiotic resistance. Therefore, it was take the advantage of this macroscopic feature of P. aeruginosa to monitor its evolution in CF environment after in vitro initial colonization. Despite its potential, colony morphology characterisation method exhibit has some restrictions, such as the unclear impact of the experimental parameters in colony morphogenesis and detection of colony diversity, variability of vocabulary and inconsistent concepts about morphoiogical traits and the time to return results. Therefore, before analysing P. aeruginosa clonal diversification in CF environment, it was attempted to introduce some improvements on this method related to standardisation of the experimental parameters and vocabulary and speed up colony analysis in three different studies. The results of the first study demonstrated that ali experimental parameters anaiysed, including colony growth time, colony density per plate, culture media, bacterial mode of growth and bacterial genetic background influenced colony morphogenesis and the detection of bacterial diversity. Therefore, a set of guidelines was created and proposed to clinical community in order to standardise the experimental parameters. Further, a morphological classification system to unambiguously characterise and describe bacterial colonies was constructed based on literature. The system was deeply tested and demonstrated to be accurate for bacterial colonies characterisation. The third study, it was aimed to introduce some quickness and highthroughput features to coiony morpholog' characterisation verifying whether MALDI-TOF MS could distinguished morphotypes. MALDI-TOF MS colony classification did not totaily match with manual colony characterisation, leading to conclude that MALDI-TOF MS provided additional information about colony variants not visible at naked eye. MALDI-TOF MS cannot thus "replace" colony morphology method but rather complement it. The data obtained in the first study demonstrated that colony morphology characterisation could be used as a reliable method to detect biofilm-derived colony morphotypes. So, it was tried to find out a colony morphology variant characteristic of biofllm lifestyle, which' could be used as a marker of P. aeruginosa bioflim growth and the development of chronic infections. Thrèsults revealed that bioflim population diversity was highly strain-dependent, hindering the unambiguous establishrrent of a coiony variant marker of P. aeruginosa biof'ilms. Nonetheless, small colony variants (SCV) demonstrated to be a strong hint of bioflim growth. In order to investigate the early adaptations undergone by P. aeruginosa after CF Iungs colonisation, bacteria grow in artificial sputum medium during ten days in the presence and absence of ciprofloxacin, an antibiotic often used in CF context. Data revealed that substantial phenotypic diversity is likely to be present in P. aeruginosa populations shortly after CF lungs colonization. Ciprofloxacin concentrations at this simulated early stage of disease demonstrated to play dose-dependent role. lnhibitory concentrations of ciprofloxacin, established based on the MIC of biofllm celI, were effective in bacterial eradication, in contrast to the sub-inhibitory doses that triggered P. aeruginosa diversification into new "fitter" variants. lmpaired swimming motility was one of the first signs of adaptation noticed in ali P. aeruginosa clonal variants, either in the presence or absence of ciprofloxacin. It was speculated that this adaptation was triggered by the environmental CF conditions due to the absence of mutators. Being so, impaired swimming motility was considered a potential disease marker for early P. aeruginosa adaptation and infection development. It was also found that some coiony morphology traits, including sheath, size and colour, could be useful indicators of antimicrobial resistance towards some antibiotics and of some virulence factors expression. During the development of this project the comparison of morphological traits and phenotypic data among colony morphotypes was a hard task. So, to easily perform this kind of analyses it was created a new ontology-based tool for the description of colony morphologies formed by bacteria, called MorphoCol. MorphoCoi establishes and standardises in consistent way the minimum information necessary to describe colony morphotypes and also the phenotypic data related to the colony-forming bacteria. In near future, this knowledgebase will transform the findings of this project and upcoming results related to colony morphology variation into valuable information for clinical decision making. The overali data will have great impact on CF disease management because relevant improvements were introduced in the colony morphology characterisation method that will strengthen clinical diagnosis. Clonai diversification in CF lungs and other host sites could be now better monitored and accurately described. Moreover, the identification of a putative CF disease marker of eariy P. aeruginosa adaptation and infection development will also assist in the design of tailored effective antimicrobial therapies.Pseudomonas aeruginosa é a maior causa das elevadas taxas de morbidez e mortalidade associada a pacientes com fibrose cística (FC). Apesar dos longos e agressivos tratamentos com antibióticos, a P. aeruginosa persiste originando infecções crónicas. A sua longa persistência deve-se a sofisticados mecanismos de adaptação, designadamente a diversificação clonal em fenótipos altamente adaptados às condições de FC. Enquanto a comunidade médica aguarda pelo desenvolvimento de terapias eficazes direcionadas ao regulador mutacional de condutância, o standard terapêutico seguido é a erradicação da P. aeruginosa logo que detetada. A erradicação atempada da P. aeruginosa evita ou, pelo menos retarda, o desenvolvimento de infecções crónicas, preservando assim a função respiratória. Contudo, para obter a erradicação bacteriana é necessário perceber os mecanismos iniciais de adaptação da P. aeruginosa para que os agentes antimicrobianos os possam bloquear. Assim sendo, este projeto pretendeu determinar quais as adaptações iniciais da P. aeruginosa após colonização das vias respiratórias de pacientes com FC e as suas respetivas forças impulsionadoras para diversas situações clínicas. As alterações na morfologia de colónia é um dos indicadores de diversificação clonal das bactérias em ambiente de FC. Estas alterações estão igualmente associadas à expressão diferencial de factores de virulência e à resistência a antibióticos. Assim, utilizou-se esta característica macroscópica da P. aeruginosa para monitorizar a sua evolução e adaptação em ambiente FC. Apesar do seu potencial, a caracterização de morfologia de colónias apresenta algumas restrições, tais como o impacto incerto dos parâmetros experimentais na morfogénese das colónias e na detecção de diversidade de colónias, a variabilidade de vocabulário e conceitos inconsistentes sobre os traços morfológicos, e o tempo de obtenção de resultados. Assim, antes da análise da diversidade clonal da P. aeruginosa em ambiente FC, tentou-se introduzir alguns melhoramentos neste método em três diferentes estudos, nomeadamente na estandardização dos parâmetros experimentais e vocabulário, bem como acelerar a obtenção de resultados. Os resultados do primeiro estudo demonstraram que todos os parâmetros experimentais analisados (tempo de crescimento das colónias, densidade de colónias por placa de cultura, meio de cultura, modo de crescimento bacteriano e os antecedentes genéticos) influenciaram a morfogénese das colónias e na detecção de diversidade bacteriana. Assim sendo, foi elaborado e proposto um conjunto de linhas de orientação para estandardização dos parâmetros experimentais. Posteriormente, foi construído um sistema de classificação para inequivocamente caracterizar a morfologia de colónias com base na literatura. Este sistema foi amplamente testado e demonstrou-se adequado para a caracterização de colónias bacterianas. No terceiro estudo tentou-se introduzir alguma celeridade e aumento do débito de resultados verificando se MALDI-TOF MS seria capaz de distinguir os moríótipos. Esta técnica forneceu uma classificação dos morfótipos ligeiramente diferente comparativamente à classificação manual. Tal, leva a concluir que MALDI-TOF MS fornece informação adicional sobre os morfótipos que não foi observável macroscopicamente. Assim, o MALDITOF MS não pode assim substituir" o método de caracterização manual de morfologia de colónias mas pode ser um método complementar. Os dados obtidos com o primeiro estudo demonstraram que a caracterização de morfologia de colónias pode ser usado na detecção de morfótipos derivados de biofilmes. Assim, tentou-se identificar um variante morfológico característico do crescimento em biofilmes que possa ser posteriormente usado para detectar este modo de crescimento e, consequentemente o desenvolvimento de infecções crónicas. Os resultados revelaram que diversidade população dos biofilmes foi dependente da estirpe, dificultando assim a identificação do variante morfológico característico de biofilme. Ainda assim, verificou-se que as small colony vanants (SCV) são um forte indicador do crescimento séssil. De forma a investigar as adaptações iniciais da P. aeruginosa após colonização de vias respiratórias com FC, as bactérias foram crescidas em muco artificial (MA) na presença e ausência de ciprofloxacina durante 10 dias. Os resultados deste estudo revelaram que é provável a existência de diversidade fenotípica logo após a colonização das vias respiratórias. As diferentes concentrações de ciprofloxacina testadas demonstraram desempenhar uma ação dose-dependente. Concentrações inibitórias, estabelecidas com base na MIC de células de biofilme, foram eficazes na erradicação da P. aeruginosa em MA, contrariamente às concentrações sub-inibitórias. Estas concentrações "acionaram" a diversificação fenotípica em variantes melhor adaptados ao ambiente FC. A limitada capacidade de swimming foi um dos sinais de adaptação inicial da P. aeruginosa verificada em todos os variantes fenótipicos. Foi assim especulado que esta adaptação foi promovida pelas condições típicas de FC devido à ausência de mutators. Como tal, a capacidade limitada de swimming foi considerada um potencial marcador dos estágios iniciais do desenvolvimento de infecções em FC. Foi igualmente verificado que alguns traço morfológicos de colónias, nomeadamente a bainha, tamanho e cor, foram indicadores de alterações na expressão de fatores de virulência e resistência a antibióticos. Durante o desenvolvimento deste projeto a comparação de traços morfológicos das colónias, bem como de dados fenótipos foi uma tarefa árdua. Neste sentido, foi desenvolvida uma base de dados, designada por MorphoCol, para a descrição de morfologia de colónias formadas por bactérias. MorphoCol estabelece e uniformiza a informação mínima necessária de uma forma estruturada e consistente. Num futuro próximo, esta base de dados transformará os resultados deste projeto e outros subsequentes em informação relevante para a tomada de decisão clínica. O conjunto de resultados obtidos neste projeto terá um impacto considerável na gestão clínica da FC devido aos consideráveis melhoramentos introduzidos no método de caracterização de colónias pois fortalecerão o diagnóstico clínico. A diversificação clonal nas vias respiratórias de FC e nos outros locais pode ser assim melhor monitorizada e descrita adequadamente. Para além disso, a identificação de possíveis marcadores para estágios iniciais de adaptação de P. aeruginosa em FC ajudarão na definição de terapias antimicrobianas eficazes

Estratégias de interação entre pais-professor-aluno na aprendizagem da viola d'arco

O presente relatório apresenta as atividades desenvolvidas ao longo da disciplina de Prática de Ensino Supervisionada, inserida no Mestrado em Ensino de Música. Dentro do nosso atual cenário educacional musical, é necessário cada vez mais envolver o meio familiar na aprendizagem instrumental do aluno. Para isso, os professores de instrumento devem investigar, compreender e utilizar estratégias de interação entre paisprofessor- aluno, de forma a agilizar o processo de aprendizagem. Com este projeto pretende-se compreender o impacto na interação entre pais-professor-aluno no ensino e aprendizagem da viola d’arco através da aplicação consciente de estratégias e que tipo de interação é gerado por elas. São duas as estratégias aplicadas: a utilização de Guiões de Estudo semanais para Alunos e Pais e a criação de um Conto Musicado, intitulado “Era uma vez uma Viola D’Arco…”, escrito por Alunos e PaisThe present dissertation presents the activities developed throughout the discipline of Supervised Teaching Practice, inserted in the Master in Music Teaching. Within our current musical educational scenario, it is increasingly necessary to involve the family environment in the student's instrumental learning. For this, the instrument teachers should investigate, understand and use strategies of interaction between parentsteacher- student, in order to streamline the learning process. This project intends to understand the impact on the interaction between parent-teacher-student in the teaching and learning of viola through the conscious application of strategies and what kind of interaction is generated by them. There are two strategies applied: the use of weekly Study Guides for Students and Parents and the creation of a Music Tale, titled "Once upon a Time, a Viola ..." written by Students and ParentsMestrado em Ensino de Músic

Specific hygiene procedures and practices assessment : a cross-sectional study in fresh fishery product retailers of Lisbon’s traditional food markets

Research Areas: Food Science & TechnologyThis study aimed to assess the compliance of specific hand hygiene and food contact surfaces hygiene in fresh fishery product retailers (FFPR) and its relation to socio-demographic characteristics that might influence the observed procedures and practices. A cross-sectional study was conducted in traditional food markets’ FFPRs (N = 74) using interviews and audits for data collection purposes. Results revealed that women were overrepresented among FFPR managers; most were over 50 years of age and had a long working experience in fish retail activities, despite a low educational level. The majority had attended training courses in food hygiene, safety, and Hazard Analysis and Critical Control Points-based procedures. Both documental assessment and hand hygiene procedures and practices verification revealed a low level of compliance. Many documents supporting hygiene practices were lacking. Several non-conforming requisites were identified related to the handwashing procedure and equipment and to glove wearing practices. A higher level of compliance was obtained in food contact surfaces hygiene procedures and practices verification, with complete hygiene procedures applied and proper cleaning of utensils and chemical products management in several cases. These findings reflect an ambiguous understanding and misconceptions related to hygiene procedures and practices, suggesting the need to improve and update FFPR food handler training regarding basic food hygiene and safety.info:eu-repo/semantics/publishedVersio

Atención plena, alimentación consciente y alimentación intuitiva en el abordaje del obesidad y de los trastornos alimentarios

Objetivo: desarrollar una revisión integrativa de la literatura sobre el papel de las intervenciones basadas en el mindfulness, mindful eating y alimentación intuitiva en el abordaje del sobrepeso / obesidad y de los trastornos alimentarios. Método: para esta revisión, se consultaron las basesde datos PubMed, PePSIC, PsycINFO, LILACS, IBECS, SciELO y BVS-Psi, y se incluyeron artículos publicados en los últimos 10 años. Después de analizar los estudios recuperados, 38 artículosconstituyeron el corpus final de la revisión. Resultados: los estudios han demostrado los beneficios las intervenciones basadas en el mindfulness, ME y alimentación intuitiva para el comportamientoalimentario y los aspectos emocionales y psicológicos de las personas con sobrepeso/obesidad y trastornos alimentarios, reduciendo la alimentación emocional, el comer basado en factores externos y en los episodios de binge eating; además de reducir los niveles de estrés, ansiedad y depresión. Para la pérdida de peso, los resultados son menos consistentes, dado que varios estudios no han encontrado diferencias para este parámetro con las intervenciones. Conclusiones: las intervenciones basadas en el mindfulness, mindful eating y alimentación intuitiva son prometedoras para el abordaje del sobrepeso/obesidad y de los trastornos alimentarios, especialmente con respecto al comportamiento alimentario y los aspectos psicoemocionales.Objective: to examine and summarize studies of mindfulness, mindful eating and intuitive eating in the approach of overweight / obesity and eating disorders. Method: for this integrative review, PubMed, PePSIC, PsycINFO, LILACS, IBECS, SciELO and BVS-Psi databases were consulted, andpublished articles published in the last 10 years. After analyzing the recovered studies, 38 articles made up the final review corpus. Results: the studies demonstrated the benefits of approaches centered on mindfulness, mindful eating and intuitive eating on eating behavior and on the emotional and psychological aspects of  verweight and eating disorders individuals, harmful or emotional, followed by external factors and compulsive eating; as well as levels of stress, anxiety and depression. For weight loss, the results are less consistent, since several studieshave found no difference for this parameter. Conclusions: as selected focused on mindfulness, mindful eating and intuitive eating are promising for addressing obesity and obesity and eating disorders, especially with regard to eating behavior and psycho-emotional aspects.Objetivo: realizar uma revisão integrativa da literatura acerca do papel das intervenções baseadas em mindfulness, mindful eating e comer intuitivo na abordagem do sobrepeso e obesidade e dos transtornos alimentares. Método: para esta revisão foram consultadas as bases PubMed, PePSIC, PsycINFO, LILACS, IBECS, SciELO e BVSPsi, e incluídos artigos publicados nos últimos 10 anos. Após o processo de análise dos estudos recuperados, 38artigos compuseram o corpus final da revisão. Resultados: os estudos demonstraram os benefícios das abordagens centradas no mindfulness, mindful eating e comer intuitivono comportamento alimentar e nos aspectos emocionais e psicológicos dos indivíduos com excesso de peso e transtorno alimentar, reduzindo o comer emocional, comer guiado porfatores externos, e episódios de binge eating; bem como reduzindo os níveis de estresse, ansiedade e depressão. Para a perda de peso, os resultados são menos consistentes, haja visto que diversos estudos não observaram diferença para este parâmetro. Conclusões: as intervenções centradas no mindfulness, mindful eating e comer intuitivo são promissoras para a abordagem do sobrepeso/obesidade e dos transtornos alimentares, especialmente no que tange ao comportamento alimentar e aos aspectos psicoemocionais

Em Casa Observamos Saúde (ECOS)’ by telephone and web: results from the 2016 first wave implementation

Em 2016 o inquérito ECOS foi implementado, através de entrevista telefónica assistida por computador, à semelhança de vagas anteriores, e ainda por via eletrónica. O questionário foi respondido por um elemento com 18 ou mais anos residente na unidade de alojamento, que prestou informação sobre a sua saúde e dos restantes elementos do agregado (por proxy). No total obtiveram-se 803 entrevistas concluídas e uma taxa de participação global de 79,9%. Considerando os contactos realizados via telefónica, a taxa de participação foi de 77,4% e de 26,5% por via web. Em ambas as vias, a maioria dos respondentes era do sexo feminino, encontrando-se a maior frequência de respondentes por telefone no grupo etário 60-69 e via web no grupo 40-49. As taxas de resposta obtidas em 2016 foram semelhantes às obtidas em outras vagas do ECOS por via telefónica (aproximadamente 80% vs 79 a 86%), e na via web foi superior às obtidas em estudos similares. O perfil dos respondentes em cada uma das vias foi consistente com o observado em estudos que utilizaram estas metodologias, nomeadamente, uma maior frequência de respondentes do sexo feminino e uma maior adesão à via web dos grupos etários mais novos.In 2016 the ECOS panel was implemented through both telephone and electronic contact. The information was obtained using computer assisted telephone and web. The questionnaire was answered by a household aged 18 or more years living in the house unit, giving information on himself and proxy to other household cohabitants. A total of 803 completed inter views were obtained which corresponds to a total participation rate of 79.9%. Considering the phone contacts the participation rate by contact type was 77.4% by phone and 26.5% by web. For both methods the highest percentages of respondents were female; the highest frequency of telephone respondents was obtained in the 60-69 age groups and by web in the 40-49 years individuals. Response rates obtained by telephone in 2016 are comparable to those obtained in other ECOS waves (80% vs 79-86%) and the rates via web were higher than those obtained in similar studies. The profile of respondents was consistent with that obser ved in studies using these methods, including increased frequency of female respondents and greater adherence to the web via the younger age groups