Health and Technology Research Center

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Publications

Algorithm to assess the presence of Aspergillus fumigatus resistant strains: the case of Norwegian sawmills

Publication . Viegas, Carla; Almeida, Beatriz; Aranha Caetano, Liliana; Afanou, Anani; Straumfors, Anne; Veríssimo, Cristina; Gonçalves, Paulo; Sabino, Raquel

Association between selection pressure caused by the use of azole fungicides in sawmills and the development of fungal resistance has been described. The aim of this study was to implement an algorithm to assess the presence of Aspergillus section Fumigati resistant strains in sawmills. Eighty-six full-shift inhalable dust samples were collected from eleven industrial sawmills in Norway. Different culture media were used and molecular identification to species level in Aspergillus section Fumigati was done by calmodulin sequencing and TR34/L98H and TR46/Y121F/T289A mutations were screened by real-time PCR assay and confirmed by cyp51A sequencing. Six Fumigati isolates were identified as A. fumigatus sensu stricto and two of these grew on azole-supplemented media and were further analyzed by real-time PCR. One was confirmed to be a TR34/L98H mutant. The obtained results reinforce the need to assess the presence of A. fumigatus sensu stricto resistant isolates at other workplaces with fungicide pressure.

2022-05Journal article

Open access

Settleable dust and bioburden in Portuguese dwellings

Publication . Viegas, Carla; Dias, Marta; Almeida, Beatriz; Vicente, Estela; Aranha Caetano, Liliana; Carolino, Elisabete; Alves, Célia

Monitoring campaigns in several buildings have shown that occupants exposed to contaminated indoor air generally exhibit diverse health symptoms. This study intends to assess settleable dust loading rates and bioburden in Portuguese dwellings by passive sampling onto quartz fiber filters and electrostatic dust cloths (EDCs), respectively. Settled dust collected by EDCs was analyzed by culture-based methods (including azole-resistance screening) and qPCR, targeting four different toxigenic Aspergillus sections (Flavi, Fumigati, Circumdati, and Nidulantes). Dust loading rates and bioburden showed higher variability in the summer season. In both seasons, Penicillium sp. was the one with the highest prevalence (59.1% winter; 58.1% summer), followed by Aspergillus sp. in winter (13.0%). Fungal contamination increased in the winter period, while bacterial counts decreased. Aspergillus sections Circumdati and Nidulantes, detected in voriconazole supplemented media, and Aspergillus sections Fumigati and Nidulantes, detected by molecular tools, were found in the winter samples. This study reinforces the importance of applying: (a) Passive sampling methods in campaigns in dwellings; (b) two different culture media (MEA and DG18) to assess fungi; (c) in parallel, molecular tools targeting the most suitable indicators of fungal contamination; and (d) azole resistance screening to unveil azole resistance detection in fungal species.

2020-11Journal article

Open access

Estratégias não-farmacológicas para redução da ansiedade em doentes submetidos a PET/CT: revisão sistematizada de literatura

Publication . Pedro, Beatriz; Henriques, Daniela; Grilo, Ana Monteiro; Vieira, Lina Oliveira

Introdução – A ansiedade, caracterizada por sentimentos de tensão e preocupação relativamente a situações que o indivíduo perceciona como potencialmente perigosas ou desconhecidas, é uma das reações emocionais experienciadas pelos doentes oncológicos aquando da realização do exame Positron Emission Tomography/Computed Tomography (PET/CT). Elevados níveis de ansiedade podem influenciar negativamente a experiência e a satisfação do paciente, bem como ter implicações a nível imagiológico. Deste modo, a avaliação da ansiedade nesses pacientes é de extrema importância, assim como a necessidade de adotar estratégias de intervenção eficazes na sua redução. Objetivos – Nesta revisão sistematizada pretende-se identificar e analisar a eficácia de técnicas não-farmacológicas que visam a redução da ansiedade em doentes submetidos a PET/CT, antes e durante o exame. Métodos – Recorreu-se a duas bases de dados (PubMed e Science Direct) e a outras fontes de pesquisa de onde resultaram 22 estudos. Foram incluídos todos os estudos que incluíssem estratégias para redução da ansiedade em exames PET/CT e que estivessem publicados nas línguas portuguesa e inglesa. Foram excluídos artigos de revisão sistemática, com população pediátrica e que não apresentassem o texto na íntegra. Resultados – Foram selecionados para análise oito estudos, num espaço temporal entre 2000 e 2020. Identificaram-se quatro estratégias que visam a redução da ansiedade nos doentes submetidos a PET/CT: música, meios audiovisuais, práticas de mindfulness e comunicação profissional de saúde-doente. Conclusão – São várias as técnicas que podem ser aplicadas de modo a minimizar o sentimento de ansiedade no doente e proporcionar uma melhor experiência no serviço. No entanto, ainda existe pouca evidência neste tema. Sendo assim, sugerem-se estudos adicionais que possam identificar mais técnicas e comprovar a sua eficácia.

2020-11Journal article

Open access

Cytotoxicity of filtering respiratory protective devices from the waste sorting industry: a comparative study between interior layer and exhalation valve

Publication . Viegas, Carla; Twarużek, Magdalena; Dias, Marta; Almeida, Beatriz; Carolino, Elisabete; Soszczyńska, Ewelina; Viegas, Susana; Caetano, Liliana Aranha

Filtering respiratory protection devices (FRPD) is mandatory for workers to wear in the Portuguese waste-sorting industry. Previous results regarding microbial contamination found on the FRPD interior layer raised the question of whether microbial contamination from the exhalation valve would also have cytotoxicity effects. Since the FRPD exhalation valves are very close to workers’ nose and mouth, they represent a source of exposure to bioburden by inhalation. This study aimed to evaluate the cytotoxicity of the microbial contamination present in the FRPD exhalation valves. For this purpose, the cytotoxicity effects were determined through the MTT assay in two different cell lines (human A549 epithelial lung cells, and swine kidney cells) and compared with previous results obtained with FRPD interior layers. The contamination present in the FRPD exhalation valves presented some cytotoxicity on epithelial lung cells, suggesting the inhalation route as a potential route of exposure through the use of FRPD in the waste-sorting industry. Half-maximal (50%) inhibitory concentration (IC50) values were lower for the FRPD interior layer than exhalation valves in lung cells, with overall cytotoxicity lower in exhalation valves when compared to the interior layer (z = −4.455, p = 0.000). Higher bacterial counts in TSA were correlated with lower IC50 values, thus, higher cytotoxicity effect in lung cells. No statistically significant differences were detected among different workplaces.

2021-10Journal article

Open access

Epigenetic and drug response modulation of epigalocaten-in-3-gallate in staphylococcus aureus with divergent resistance phenotypes

Publication . Mira, Ana Rita; Zeferino, Ana Sofia; Inácio, Raquel; Delgadinho, Mariana; Brito, Miguel; Calado, Cecília; Ribeiro, Edna

Healthcare-associated methicillin-resistant Staphylococcus aureus infections represent extremely high morbidity and mortality rates worldwide. We aimed to assess the antimicrobial potential and synergistic effect between Epigalocatenin-3-gallate (EGCG) and different antibiotics in S. aureus strains with divergent resistance phenotypes. EGCG exposure effects in epigenetic and drug resistance key modulators were also evaluated. S. aureus strains (n = 32) were isolated from infected patients in a Lisbon hospital. The identification of the S. aureus resistance phenotype was performed through automatized methods. The antibiotic synergistic assay was performed through disk diffusion according to EUCAST guidelines with co-exposure to EGCG (250, 100, 50 and 25 µg/mL). The bacteria’s molecular profile was assessed through FTIR spectroscopy. The transcriptional expression of OrfX, SpdC and WalKR was performed by using qRT-PCR. FTIR-spectroscopy analysis enabled the clear discrimination of MRSA/MSSA strains and the EGCG exposure effect in the bacteria’s molecular profiles. Divergent resistant phenotypes were associated with divergent transcriptional expression of the epigenetic modulator OrfX, particularly in MRSA strains, as well as the key drug response modulators SpdC and WalKR. These results clearly demonstrate that EGCG exposure alters the expression patterns of key epigenetic and drug response genes with associated divergent-resistant profiles, which supports its potential for antimicrobial treatment and/or therapeutic adjuvant against antibiotic-resistant microorganisms.

2023-03-04Journal article

Open access