Assessing fungal burden and its implications for health in an elderly care centre: a comprehensive study

Author gratefully acknowledges the FCT/MCTES national support through the UIDB/05608/2020, and UIDP/05608/2020 This work is also supported by national funds through FCT/MCTES/FSE/UE, 2023.01366.BD; UI/BD/153746/2022, and CE3C unit UIDB/00329/2020, UI/BD/151431/2021, and Instituto Politécnico de Lisboa, national support through IPL/2022/InChildhealth/12M, IPL/IDI&CA2023/FoodAIIEU_ESTeSL, IPL/IDI&CA2023/ASPRisk_ESTeSL, and IPL/IDI&CA2023/ARAFSawmills_ESTeSL.Introduction: Protection thresholds and reference conditions for indoor air pollution, in commercial and service buildings and assessment methodology, leaving Elderly Healthcare facilities neglected for such parameters. The study aims to assess the fungal burden in an elderly care center and its health implications. Target groups: workers, and elderly residents.info:eu-repo/semantics/publishedVersio

Expression of mTOR in normal and pathological conditions

The H&TRC authors gratefully acknowledge the FCT/MCTES national support through the UIDB/05608/2020 and UIDP/05608/2020. This work was part of the IPL/2021/GATumor_ESTeSL project.The mechanistic/mammalian target of rapamycin (mTOR), a protein discovered in 1991, integrates a complex pathway with a key role in maintaining cellular homeostasis. By comprising two functionally distinct complexes, mTOR complex 1 (mTORC1) and mTORC2, it is a central cellular hub that integrates intra- and extracellular signals of energy, nutrient, and hormone availability, modulating the molecular responses to acquire a homeostatic state through the regulation of anabolic and catabolic processes. Accordingly, dysregulation of the mTOR pathway has been implicated in a variety of human diseases. While major advances have been made regarding the regulators and effectors of the mTOR signaling pathway, insights into the regulation of mTOR gene expression are beginning to emerge. Here, we present the currently available data regarding the mTOR expression regulation at the level of transcription, translation, and mRNA stability and systematize the current knowledge about the fluctuations of mTOR expression observed in several diseases, both cancerous and non-cancerous. In addition, we discuss whether mTOR expression changes can be used as a biomarker for diagnosis, disease progression, prognosis, and/or response to therapeutics. We believe that our study will contribute to the implementation of new disease biomarkers based on mTOR as it gives an exhaustive perspective on the regulation of mTOR gene expression in both normal and pathological conditions.info:eu-repo/semantics/publishedVersio

Sampling elementary schools: a comprehensive approach for microbial indoor air quality assessment

This work is supported by national funds through FCT/MCTES/FSE/UE, 2023.01366.BD; UI/BD/153746/2022 and CE3C unit UIDB/00329/2020 (https://doi.org/10.54499/UIDB/00329/2020); UI/BD/151431/2021 (https://doi.org/10.54499/UI/BD/151431/2021); and Instituto Politécnico de Lisboa, national support through IPL/2022/InChildhealth/BI/12M; IPL/IDI&CA2023/FoodAIIEU_ESTeSL; IPL/IDI&CA2023/ASPRisk_ESTeSL; and IPL/IDI&CA2023/ARAFSawmills_ESTeSL.Indoor air quality (IAQ) is an important determinant of children’s health since children spend around 90 % of their time in the indoor environment. In schools, children may be exposed to bioaerosols (such as viruses, bacteria, and fungi), in their classrooms, libraries, canteen, gymnasiums/sports hall, and toilets. Portugal only established protection thresholds and reference conditions for indoor air pollution in commercial and service buildings and assessment methodology through Portaria n.º 138-G/2021, neglecting schools as a specific indoor environment. This project aims to apply a multi-approach protocol (sampling and assays), to assess microbial contamination (bacteria and fungi) in Portuguese elementary schools and to suggest guidance for exposure assessors from the field to the lab.Partly funded by the European Union. Views and opinions expressed are however those of the author(s) only and do not necessarily reflect those of the European Union, the Swiss State Secretariat for Education, Research and Innovation (SERI), or the United Kingdom Research and Innovation (UKRI), or the Australian National Health & Medical Research Council (NHMRC). Neither the European Union nor the granting authorities can be held responsible for them.info:eu-repo/semantics/publishedVersio

Characterization of microbiological contamination in Portuguese elementary schools

H&TRC authors gratefully acknowledge the FCT/MCTES national support through the UIDB/05608/2020 and UIDP/05608/2020.Ninety percent of children's time is spent in an indoor environment. Most school-age children’s exposure occurs in schools, at home, in sports halls, and commuting. IAQ is an important determinant of human health, especially for children. Project goals: characterization of air quality in children’s environments; identification of emission sources and influencing factors affecting children's exposure; estimation of associated health effects resulting from exposures; development of novel technologies to improve indoor environmental quality to reduce health effects; engaging school children, educational experts, and policymakers in the scientific process; preparation of quality standards and guidelines regarding IAQ.info:eu-repo/semantics/publishedVersio

mTOR expression regulation and new anticancer applications for apigenin

Mestrado em Tecnologias Clínico-LaboratoriaisA apigenina (4',5,7-tri-hidroxiflavona) é um dos flavonoides mais abundantes do mundo vegetal e um dos fenólicos mais pesquisados. Sabe-se que a apigenina reduz a proliferação celular nas células de cancro gástrico, mas resta saber se a apigenina reduz o mTOR nas células de cancro gástrico, se esta redução no mTOR é de alguma forma responsável pela supracitada diminuição da proliferação, e se a exposição à apigenina é total ou parcialmente responsável por esta diminuição. A via de sinalização do mTOR encontra-se hiperactivada em cerca de 70% dos cancros humanos, nomeadamente no cancro gástrico, assim como em doenças muito prevalentes na nossa população como a diabetes e as doenças cardiovasculares. mTOR é um centro celular que integra sinais intra e extracelulares em termos de energia, nutrientes, e disponibilidade hormonal, modulando as respostas moleculares para adquirir um estado homeostático através da regulação de processos anabólicos e catabólicos. Consequentemente, a desregulação da via mTOR tem sido associada a uma variedade de doenças humanas. Embora tenham surgido nos últimos anos grandes avanços relativamente aos reguladores e efeitos da via de sinalização do mTOR, pouco se sabe sobre a regulação da expressão genética do mTOR. Os dados atualmente disponíveis sobre as alterações de expressão de mTOR observadas em várias doenças, particularmente cancros humanos, são aqui apresentados, e o conhecimento atual sobre a regulação de mTOR a nível transcricional e translacional é apresentado de forma sistematizada e é descrito como diferentes miRNAs afetam a sinalização de mTOR em condições patológicas. Além disso, utilizando linhas celulares de cancrogástrico Gp202, foi possível caracterizar o potencial anti-tumorigénico da apigenina, descrevendo a sua influência na morfologia celular e nos mecanismos associados à morte celular. Este estudo ajudará na prossecução da investigação das propriedades anti-tumorais da apigenina em relação ao cancro gástrico, particularmente o seu possível efeito sobre o mTOR, bem como no desenvolvimento de novos inibidores do mTOR, uma vez que proporciona uma perspetiva exaustiva sobre a regulação da expressão do gene mTOR.ABSTRACT - Apigenin (4',5,7-trihydroxyflavone) is one of the most abundant flavonoids in the plant world and one of the most researched phenolics. Apigenin is known to reduce cell proliferation in gastric cancer cells, but it remains to be seen whether apigenin reduces mTOR in gastric cancer cells, whether this reduction in mTOR is in any way responsible for the aforementioned decrease in proliferation, and whether exposure to apigenin is fully or partially responsible for this decrease. The mechanistic/mammalian target of rapamycin (mTOR) signaling pathway is hyperactivated in about 70% of human cancers, including gastric cancer, as well as in diseases that are very prevalent in our population such as diabetes and cardiovascular disease. mTOR is a central cellular hub that integrates intra- and extracellular signals in terms of energy, nutrient, and hormone availability, modulating the molecular responses to acquire a homeostatic state through the regulation of anabolic and catabolic processes. Accordingly, dysregulation of the mTOR pathway has been implicated in a variety of human diseases. While major advances have emerged regarding the regulators and effects of the mTOR signaling pathway, little is known about the regulation of mTOR gene expression. The currently available data regarding mTOR expression changes observed in several diseases, particularly human cancers, is presented here, and the current knowledge about mTOR regulation at the transcriptional and translational levels is systematized. It also demonstrated how different miRNAs affect mTOR signaling in pathological conditions. Furthermore, utilizing Gp202 gastric carcinoma cell lines, it was possible to define apigenin's anti-tumor potential by establishing its influence on cell morphology and the mechanisms associated with cell death. This study, I believe, will assist in the pursuit of research into apigenin's anti-tumoral properties in relation to gastric cancer, particularly its possible effect on mTOR, as well as in the development of new mTOR inhibitors since it provides an exhaustive perspective on the regulation of mTOR gene expression.N/

Assessing the impact of climate change on indoor fungal contamination in Lisbon Metropolitan Area primary schools: a comprehensive study

H&TRC authors gratefully acknowledge FCT/MCTES national support through the 2023.01366.BD; UIDB/05608/2020; UIDP/05608/2020, and IPL/2022/InChildhealth/BI/12M. The ESTeSL- Escola Superior de Tecnologia e Saúde de Lisboa, Instituto Politécnico de Lisboa, national support through IPL/IDI&CA2023/FoodAlleEU; IPL/IDI&CA2023/ASPRisk, and IPL/IDI&CA2023/ARAFSawmills.The increasing occurrence of severe weather events caused by global climate change raises concerns about indoor fungi. These changes can potentially alter fungal communities, promoting resistant strains, and intensifying mycotoxin production, having significant implications for human health. In educational settings, inadequate ventilation and high moisture levels amplify indoor fungal growth and mycotoxin contamination, posing a health risk that contributes to respiratory illnesses and allergic reactions in children and school staff. Our research explores the complex relationship between climate change and fungal diseases, highlighting the importance of temperature and moisture in driving fungal growth and mycotoxin production. This project focuses on the Lisbon metropolitan area and aims to assess the indoor levels of contamination and human exposure to azole-resistant fungi and mycotoxins in primary schools. The objective of this investigation is to carry out a comprehensive microbial characterization of the exposure to fungal contamination in primary educational institutions and to achieve that goal a comprehensive sampling campaign was employed, by the use of active and passive sampling methods and material collection from schools. Air samples utilizing the MAS-100 device, which collected 400L at a flow rate of 200 L/min, and the Anderson six-stage device, which collected 200L at a flow rate of 28.3 L/min were performed. Additionally, we collected samples of mops and surface swabs, and samples of settled dust through vacuuming and EDC, located in the sampling sites at 1.5 m height for 30 days, and collected during approximately 30 days. The samples will be analysed by culture-based methods, through the inoculation onto two different culture media: malt extract agar (MEA) supplemented with chloramphenicol (0.05%) incubated at 27ºC and dichloran-glycerol agar (DG18), incubated at 27ºC and 37ºC for 6 days. We will also be performing molecular detection of the selected fungal sections (Aspergillus sections Circumdati, Flavi, Fumigati, and Nidulantes). To evaluate the degree of mycotoxin contamination in school environments, we will employ high-performance liquid chromatography (HPLC) to detect the presence of 38 different mycotoxins. The preliminary results that will be presented are specifically related to the active air sampling. The ultimate objective is to provide essential insights for the development of effective risk management strategies that safeguard the health of school children and personnel in the face of growing fungal threats.info:eu-repo/semantics/publishedVersio

Microbial contamination of bedding material: one health in poultry production

FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.In poultry farms, the mixture of bedding material, chicken excrement, and feathers seems to play an important role in pathogen development which may contribute to a potential risk of zoonosis, spreading the disease through the food chain. The purpose of this study was to analyze microbial contamination in bedding material and other matrices as well as potential antimicrobial resistances in chicken production facilities and also to identify the sampling techniques and assays used. This study evidences the available data published, following the PRISMA methodology. Among the environmental samples, surface swabs were frequently used as a passive sampling technique. Morphological identification was performed in all studies. Of all the matrices, the bedding material was the most contaminated. Most studies focused on bacterial contamination, with Salmonella sp. and Campylobacter sp. being commonly reported and three studies evidenced fungal contamination, being Penicillium sp.- and Aspergillus sp.-dominant. Mycotoxin assessment was only performed in one study, being identified in all bedding samples. The screening for bacteria resistance evidenced bacteria multidrug resistance; however, fungal susceptibility to azoles was not assessed in any of the analyzed studies. Briefly, this review evidences microbial contamination in poultry facilities, emphasizing animals’ bedding as a potential source of contamination. Additionally, this study contributes to a sampling and analysis protocol proposal to assess microbial contamination in this setting. Additionally, the knowledge gaps identified highlight the need for further research regarding microbial contamination and toxicological potential on animals’ bedding in order to mitigate exposure in poultry pavilions.info:eu-repo/semantics/publishedVersio

Insights of Portugal’s teaching staff microbial occupational exposure

This project was supported by FCT/MCTES UIDP/05608/2020, and UIDB/05608/2020. This work is also supported by national funds through FCT/MCTES/FSE/UE, 2023.01366.BD; UI/BD/153746/2022 and CE3C unit UIDB/00329/2020; UI/BD/151431/2021; and Instituto Politécnico de Lisboa, national support through IPL/2022/InChildhealth/BI/12M; IPL/IDI&CA2023/FoodAIIEU_ESTeSL; IPL/IDI&CA2023/ASPRisk_ESTeSL; and IPL/IDI&CA2023/ARAFSawmills_ESTeSL.EU strategies in education focus on risk factor awareness to promote a safe and healthy environment for the community. Health and safety promotion faces challenges due to the lack of adjusted policies for teaching staff. Teachers may be exposed to biohazards in schools, posing a health risk, which remains unexplored. Effective preventive measures and risk management actions require a thorough risk assessment. This study aims to conduct a microbial assessment regarding school staff personal exposure to bacterial and fungal loads.Partly funded by the European Union’s Horizon 2021 research and innovation program under grant agreement no. 101056883 and received co-founding from the author’s organizations and/or ministries. This work has received funding from the Swiss State Secretariat for Education, Research, and Innovation (SERI) grant no. 22.00324, from the United Kingdom Research and Innovation (UKRI) grant no. 10040524, and the Australian National Health & Medical Research Council (NMHRC) grant no. APP2017786 and APP2008813.info:eu-repo/semantics/publishedVersio

First insights of Portuguese primary schools’ fungal assessment: is indoor air quality complying with Portuguese legal framework?

H&TRC authors gratefully acknowledge FCT/MCTES national support through the 2023.01366.BD; UIDB/05608/2020; UIDP/05608/2020, and IPL/2022/InChildhealth/BI/12M. The ESTeSL- Escola Superior de Tecnologia e Saúde de Lisboa, Instituto Politécnico de Lisboa, national support through IPL/IDI&CA2023/FoodAlleEU; IPL/IDI&CA2023/ASPRisk and IPL/IDI&CA2023/ARAFSawmills.The assessment of Microbial Indoor Air Quality in elementary schools is essential to create healthier school environments and improve children's health outcomes. Portugal only established protection thresholds for microbial exposure, in commercial and service buildings through Portaria nº 138-G/2021, leaving schools neglected for such parameters. Since this legislation has already been proven insufficient for fungal parameters (based on indoor/outdoor ratio) in other critical environments such as healthcare centers, this study aims to assess fungal load compliance in different sites of schools located in the Lisbon area. An active sampling method was applied by collecting air through a MAS-100 device collecting 200L at a flow rate of 100 L/min. Of the 10 schools assessed in a summer campaign, 9 did not comply with the Portuguese legal framework in at least one site (8 out of 10 in the classrooms, 5 out of 7 in the bathroom, 4 out of 9 in the canteen, 4 out of 6 in the gymnasiums, and 3 out of 8 in the library). A critical assessment was performed to identify fungal species listed in this legislation. Although one school complies with the quantitative cut-off (I/O), critical species such as Aspergillus section Circumdati (Aspergillus ochraceus), and Aspergillus section Fumigati (Aspergillus fumigatus) present in this school do not meet the toxigenic species quantitative cut-off. The risk of exposure to pathogenic fungi poses a major public health threat for school-age children impacting not only treatable fungal infections but also students' learning conditions and outcomes.info:eu-repo/semantics/publishedVersio

Development of an indexed score to identify the most suitable biological material to assess SARS-CoV-2

No funding was directly required to develop this research. H&TRC authors gratefully acknowledge the FCT/MCTES national support through the UIDB/05608/2020 and UIDP/05608/2020.Introduction: The rapidly contagious process of respiratory viruses such as SARS-CoV-2 makes it urgent to multiply testing for diagnostics to identify the active viral shedding cases (current infection, carriage state, or, residual viral RNA) and decrease the risk of transmission to other patients and healthcare professionals. Although nasopharyngeal swabs (NPSs) are the most common specimen type used for COVID-19 diagnosis, they require supervision by a professional, and concerns have been raised regarding healthcare personnel exposure, difficulty in collection, and patient discomfort. Viral RNA can also be detected in specimens such as saliva, blood, bronchoalveolar lavage fluid, sputum, faeces, and urine. This study aimed to provide updated information about the most suitable biological material to diagnose SARS-CoV-2, considering the risk assessment, specialization needed, test cost, complexity of the collection, and sample treatment associated with the different types of specimens. Methods: An extensive search of scientific review articles were made to collect information about the biological specimens to identify SARS-CoV-2 in the urine, sputum, nasopharyngeal, oropharyngeal, bronchoalveolar (BAL), saliva, faeces, and blood. For this purpose, an index score was developed based on seven categories: Materials and Equipment; Infection Risk for the Health Professional; Infection Risk for the Patient; Collection; Cost; Specialized HR; and RNA Extraction Type. Results and Discussion: Each criterion from the index score was quoted from 1.0 to 5.0, and a sum was made to classify which specimen is the best choice to diagnose SARS-CoV-2, according to the chosen parameters. Data indicated that urine specimens are the most elementary biological sample to access. Regarding RNA extraction, NPSs, OPS, and BAL presented the maximum score. However, BAL has the lowest score regarding associated costs. Concerning sputum and saliva, all the aspects were evaluated with a score of 5.0 except for the RNA Extraction Type in sputum. Regarding the total scores of the multiple specimens, the lowest corresponds to BAL with a score of 1.7, followed by blood with 3.1 and NPSs and OPS with 3.6 and 3.7, respectively. Urine and faeces have the same value, 4.4, sputum has 4.9, and the highest and maximum possible value corresponds to saliva with 5.0, making this last specimen the most suitable for all considered parameters. Conclusion: Although OPS and NPSs are the most used specimens, there are better alternatives. Among all the specimens of the respiratory system, saliva is the most cost-effective specimen for performing SARS-CoV-2 diagnosis. Even though these infections are usually diagnosed clinically based on symptoms and local epidemiology, identification of the specific pathogen may affect clinical management and be crucial for containing potential outbreaks.info:eu-repo/semantics/publishedVersio