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

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/

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

Commercial tea samples: a natural health ally or an Aspergillus infusion

Project "Resistant mycobiota and mycotoxigenic profile of tea and medicinal plants” (IPL/2020/TEAResMyc_ESTeSL).FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.Best poster award.Introduction and Purpose - Tea consumption has been increasing in the last few years. The lack of specific legislation concerning microbial contamination of teas and infusions might represent a risk for consumers’ health. This study characterizes fungal contamination and azole resistance on green and black tea and other infusions commercially available in Portugal.info:eu-repo/semantics/publishedVersio

a review

Funding Information: Funding: This research was funded by national funds through the FCT—Fundação para a Ciência e Tecnologia, I.P., within the scope of the PhD Grant UI/BD/151431/2021. Funding Information: This research was funded by national funds through the FCT?Funda??o para a Ci?ncia e Tecnologia, I.P., within the scope of the PhD Grant UI/BD/151431/2021. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.The composition of airborne microflora in sawmills may vary to a great degree depending on the kind of timber being processed and the technology of production being used. Cases of allergy alveolitis and asthma have been reported in woodworkers who were exposed to wood dust largely infected with microorganisms. The aim of this review article is to identify studies where the microbial occupational exposure assessment was performed in sawmills and the characteristics of the contamination found, as well as to identify which sampling methods and assays were applied. This study reports the search of available data published regarding microbial occupational exposure assessment in environmental samples from sawmills, following the Preferred Reporting Items for Systematic Reviews (PRISMA) methodology. The most used sampling method was air sampling, impaction being the most common method. Regarding analytical procedures for microbial characterization, morphological identification of fungi and bacteria was the most frequent approach. Screening for fungal susceptibility to azoles was performed in two studies and four studies applied molecular tools. Regarding microbial contamination, high fungal levels were frequent, as well as high bacteria levels. Fungal identification evidenced Penicillium as the most frequent genera followed by Aspergillus sp. Mycotoxins were not assessed in any of the analyzed studies. Microbial occupational exposure assessment in sawmills is crucial to allow this risk characterization and management.publishersversionpublishe

Are we neglecting the importance of azole resistance screening in indoor air quality assessments?

Project “Waste Workers’ Exposure to Bioburden in the Truck Cab during Waste Management - W2E Bioburden” (IPL/2016/W2E_ESTeSL).Project “Occupational exposure of ambulance drivers to bioburden” (IPL/2020/BIO-AmbuDrivers_ESTeSL).FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.Best poster awardIntroduction: The microbial exposure, as well as their metabolic products, has been frequently reported as a high risk of respiratory effects and general health hazards. Regarding the Portuguese legislation for IAQ, the microbial assessment covers only air sampling and culture-based methods (malt extract agar as only culture media to be applied). Fungal legal criteria consist of a fungal load ratio between indoor and outdoor, which should be lower than 1 (I/O<1). Purpose: Analyze the importance of including the characterization of the azole resistance profile in indoor air quality assessments.info:eu-repo/semantics/publishedVersio

a systematic review

Funding Information: This work was supported by national funds through FCT/MCTES/FSE/UE, UI/BD/153746/2022, and CE3C unit UIDB/00329/2020 within the scope of a PhD grant. H&TRC authors gratefully acknowledge the FCT/MCTES national support through UIDB/05608/2020 and UIDP/05608/2020. Publisher Copyright: © 2023 by the authors.This study reports the search of available data published regarding microbial occupational exposure assessment in poultries, following the PRISMA methodology. Air collection through filtration was the most frequently used. The most commonly used passive sampling method was material collection such as dust, cages, soils, sediment, and wastewater. Regarding assays applied, the majority of studies comprised culture-based methods, but molecular tools were also frequently used. Screening for antimicrobial susceptibility was performed only for bacteria; cytotoxicity, virological and serological assays were also performed. Most of the selected studies focused on bacteria, although fungi, endotoxins, and β-glucans were also assessed. The only study concerning fungi and mycotoxins reported the carcinogenic mycotoxin AFB1. This study gives a comprehensive overview of microbial contamination in the poultry industry, emphasizing this setting as a potential reservoir of microbial pathogens threatening human, animal, and environmental health. Additionally, this research helps to provide a sampling and analysis protocol proposal to evaluate the microbiological contamination in these facilities. Few articles were found reporting fungal contamination in poultry farms worldwide. In addition, information concerning fungal resistance profile and mycotoxin contamination remain scarce. Overall, a One Health approach should be incorporated in exposure assessments and the knowledge gaps identified in this paper should be addressed in further research.publishersversionpublishe

Portuguese groceries: a potential indoor environment to be used as public health sentinel for fungal and mycotoxins contamination

This study aimed to characterize microbial contamination in Portuguese grocery stores. The research was carried out in 15 grocery stores in Cascais, Portugal. Electrostatic dust cloths (EDC) and surface swabs were used on 3 sampling sites: checkout, fruits/vegetables, and warehouse/dispenser. Fungal contamination was characterized by culture-based methods, through the inoculation onto two different culture media: malt extract agar (MEA) supplemented with chloramphenicol (0.05 %) and dichloran-glycerol agar (DG18). Screening of azole resistance was performed by inoculation of EDC extracts' in azole-supplemented Sabouraud dextrose agar (SDA) media, according to EUCAST guidelines. A total of 39 samples were screened for mycotoxin contamination. The highest fungal contamination was obtained from fruits/vegetables in swabs (76% MEA; 71% DG18). The most prevalent genera in swabs was Cladosporium sp. (54% MEA). Regarding EDC, besides Penicillium sp. (63%) the most prevalent were Aspergillus sections Circumdati (25%) and Aspergilli (35%). Penicillium sp. was highly prevalent in the checkout (61% SDA; 87% voriconazole), and warehouse/dispenser sectors (73% voriconazole; 52% posaconazole). Circumdati and Fumigati were the most prevalent Aspergillus sections in fruits/vegetables (73% and 64% in voriconazole, respectively). Four mycotoxins (fumonisin B1, B2, B3) were detected simultaneously in most of the 36 samples. Identifying the most critical workplaces in groceries concerning fungal and mycotoxin contamination is of utmost importance to human health (workers and consumers) when using a One Health approach.info:eu-repo/semantics/publishedVersio

Unveiling this Portuguese occupational exposure scenario

Copyright © 2022 Elsevier Ltd. All rights reserved.Previous studies anticipated that microorganisms and their metabolites in waste will increase as a consequence of a decreased collection frequency and due to differences in what kind of waste is bagged before collection leading to an increased exposure of workers handling the waste. This study aim was to investigate the microbial contamination present in the waste collection trucks (WCT) and in the support facilities (waste collection station - WCS). It was applied a multi-approach protocol using active (air sampling by impingement and impaction) and passive (surface swabs, electrostatic dust cloths and settled dust) sampling methods. The screening of azole-resistance, the investigation of mycotoxins and the assessment of the elicited biological responses in vitro were also carried out aiming recognizing the possible health effects of waste collection drivers. SARS-CoV-2 detection was also performed. In WCS only air samples had contamination in all the four sampling sites (canteen, operational removal core, operational removal center, and administrative service). Among all the analyzed matrices from the WCT a higher percentage of total bacterial counts and Gram-was detected in swabs (66.93%; 99.36%). In WCS the most common species were Penicillium sp. (43.98%) and Cladosporium sp. (24.68%), while on WCT Aspergillus sp. (4.18%) was also one of the most found. In the azole resistance screening Aspergillus genera was not observed in the azole-supplemented media. SARS-CoV-2 was not detected in any of the environmental samples collected, but Aspergillus section Fumigati was detected in 5 samples. Mycotoxins were not detected in EDC from WCS, while in WCT they were detected in filters (N = 1) and in settled dust samples (N = 16). In conclusion, our study reveals that a comprehensive sampling approach using active and passive sampling (e.g. settled dust sampling for a representative mycotoxin evaluation) and combined analytic methods (i.e., culture-based and molecular) is an important asset in microbial exposure assessments. Concerning the waste collection exposure scenario, the results of this study unveiled a complex exposure, particularly to fungi and their metabolites. Aspergillus section Fumigati highlight the significance of targeting this section in the waste management industry as an indicator of occupational health risk.publishersversionpublishe

Microbial contamination in the coffee industry: an occupational menace besides a food safety concern?

FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.Respiratory abnormalities among workers at coffee roasting and packaging facilities have already been reported; however, little is known about microbiological contamination inside coffee production facilities. This study intends to assess the microbial contamination (fungi and bacteria) in two coffee industries in Brazil with a multi-approach protocol for sampling and for subsequent analyses using four main sources of samples: filtering respiratory protection devices (FRPD) used by workers, settled dust, electrostatic dust cloths (EDC) and coffee beans. The fungal contamination in the assessed industries was also characterized through the molecular detection of toxigenic species and antifungal resistance. Total bacteria contamination presented the highest values in FRPD collected from both industries (7.45 × 104 CFU.m−2; 1.09 × 104 CFU.m−2). Aspergillus genera were widespread in all the environmental samples collected and sections with clinical relevance (Fumigati) and with toxigenic potential (Nigri and Circumdati) were recovered from FRPD. Circumdati section was observed in 4 mg/mL itraconazole. Sections Circumdati (EDC, coffee beans, and settled dust) and Nidulantes (EDC, coffee beans, and FRPD) were detected by qPCR. Some of the targeted Aspergillus sections that have been identified microscopically were not detected by qPCR and vice-versa. Overall, this study revealed that microbial contamination is a potential occupational risk in the milling stage and should be tackled when assessing exposure and performing a risk assessment. In addition, a multi-sampling campaign should be the approach to follow when assessing microbial contamination and FRPD should be included in this campaign. Occupational exposure to mycotoxins should be considered due to high fungal diversity and contamination. A One Health approach should address these issues in order to prevent the consumption of coffee crops and beans infected by fungi and, more specifically, to avoid widespread azole resistance.H&TRC—Health & Technology Research Center, ESTeSL - Escola Superior de Tecnologia e Saúde, Instituto Politécnico de Lisboa.info:eu-repo/semantics/publishedVersio