Proteómica cuantitativa e inmunoproteómica dirigidas al estudio de la interacción entre el hospedador y Candida albicans: respuesta mediada por macrófagos y por anticuerpos

Tesis inédita de la Universidad Complutense de Madrid, Facultad de Farmacia, Departamento de Microbiología y Parasitología, leída el 05-11-2019Candida albicans forma parte de la microbiota de individuos sanos. Sin embargo, ante un desequilibrio de la microbiota o inmunosupresión, se puede desarrollar una infección, siendo la candidiasis invasora (IC) un problema clínico relevante. Por esto, el estudio de la interacción hospedador-patógeno es crucial para conocer cómo el sistema inmunitario responde a Candida.Los macrófagos son células inmunes implicadas en el reconocimiento, fagocitosis y destrucción del hongo. Además, se ha descrito la producción de anticuerpos frente a proteínas de C. albicans durante estas infecciones. La proteómica proporciona información de la abundancia de proteínas y de las modificaciones postraduccionales durante la interacción patógeno-hospedador. Así mismo, el análisis de las proteínas secretadas por C. albicans también es importante debido al papel que tienen éstas en la interacción con el hospedador. El estudio de estas proteínas mediante inmunoproteómica resulta útil para el descubrimiento de biomarcadores para el diagnóstico de las IC.Candida albicans can be part of the microbiota of healthy individuals. However, if the balance of the normal microbiota is disrupted or the immune defences are compromised a scenario of infection can arise. Invasive candidiasis (IC) is an important health-care associated fungal infection. Therefore, the study of host –pathogen complex interplay is needed to improve the knowledge on how the immune system responds to Candida.Macrophages are key immune cells involved in recognition, phagocytosis and killing of the fungus. The production of antibodies against several C. albicans proteins has also been described during these infections.Proteomics can be used to understand host-pathogen interactions and give information on the protein abundance and on their post translational modifications. Moreover, C. albicans secreted proteins analysis is crucial due to the role of these proteins in interaction with the host and their study using immunoproteomics is useful for the discovery of biomarkers for the diagnosis of IC...Fac. de FarmaciaTRUEunpu

Incorporation of Candida albicans cell wall surface proteins into lipid vesicles : physicochemical characterization and in vitro cytotoxicity

Dissertação de mestrado em Genética MolecularOver the past two decades, Candida species have become the fourth most common cause of bloodstream infections with mortality rates higher than 50%. A significant improvement has been made in chemotherapy for candidiasis with the availability of new azole derivatives and inhibitors of glucan synthase. However, the mortality rate for invasive candidiasis has remained stable. These observations underline the urgent need for the development of novel approaches to fight fungal infections. Candida albicans cell wall has in its composition cell wall surface proteins (CWSPs), which are important targets for the host immune system. Therefore, the present study was developed in order to compare two distinct lipid systems, one constituted by phosphatidylethanolamine (PE) and another composed by DODAB:MO, regarding their ability to incorporate antigenic CWSPs for a future development of a safe and effective vaccine against C. albicans. After the successful incorporation of the CWSPs into lipid vesicles three characteristics were analyzed: average size of the lipid-CWSPs vesicles, their polydispersity index (PDI) and incorporation efficiency (IE). The PE-CWSPs vesicles presented average sizes between 268.9 nm and 492.4 nm and PDI values around 0.6. The MO, a neutral lipid, was included in the PE vesicles and the average size increased to 625.3 nm with PDI values of 0.5. DODAB:MOCWSPs vesicles average size ranged between 250.9 nm to 732.5 nm and the PDI values were considerably lower, around 0.3 to 0.5. After optimization of the delipidation method IE was determined and PE-CWSPs presented values ranging from 16.5% to 42.3%, whether PE:MOCWSPs vesicles presented lower values, around 25%. The DODAB:MO-CWSPs vesicles presented higher IE values, between 44.4% and 50,5%. Three formulations, PE (0.6mM)- CWSPs, DODAB:MO (0.1mM)-CWSPs and DODAB:MO (0.3mM)-CWSPs were chosen, from a total of eight, according to its size, PDI value and incorporation efficiency, to be tested for in vitro cytotoxicity and no significant cytotoxicity was observed. With the information collected in this work both systems, PE-CWSPs and DODAB:MOCWSPs, were considered to be suitable for the development of a vaccine against C. albicans infections.Ao longo das últimas décadas, as espécies do género Candida tornaram-se a quarta causa mais frequente de infeções sistémicas com taxas de mortalidade superiores a 50%. A quimioterapia contra candidíase tem melhorado significativamente com a disponibilização de novos derivados azólicos e inibidores da sintase dos glucanos. Contudo, a taxa de mortalidade de candidíase invasiva tem-se mantido estável. Estes estudos apontam para a necessidade urgente de desenvolver novas estratégias contra as infeções fúngicas. A parede celular de C. albicans contem proteínas à sua superfície (CWSPs), que são alvos importantes para o sistema imune. Desta forma, este estudo foi desenvolvido no sentido de comparar dois sistemas lipídicos distintos, um constituído por fosfatidiletanolamina (PE) e outro composto por DODAB:MO, no que respeita à capacidade para incorporar as CWSPs, para o futuro desenvolvimento de uma vacina segura e eficiente contra C. albicans. Após incorporação das CWSPs nas vesículas lipídicas foram analisadas três características: o tamanho médio das vesículas lipídicas, o seu índice de polidispersividade (PDI) e a sua eficiência de incorporação (IE). As vesículas PE-CWSPs apresentaram tamanhos médios entre 268,9 nm e 492,4 nm e valores de PDI entre 0,6. O lípido neutro MO foi incluído nas vesículas de PE e o tamanho destas aumentou para 625,3 nm com valores de PDI à volta de 0,5. As vesículas de DODAB:MO-CWSPs apresentaram tamanhos entre 250,9 nm e 732,5 nm e valores de PDI consideravelmente mais baixos, entre 0,3 e 0,5. Depois da otimização do método de remoção dos interferentes lipídicos foi determinada a IE e as vesículas de PECWSPs apresentaram valores entre 16,5% e 42,3% enquanto que as vesículas de PE:MOCWSPs apresentaram valores mais baixos, 25%. As vesículas de DODAB:MO-CWSPs apresentaram valores de IE mais elevados, entre 44% e 50,5%. De um total de oito formulações foram escolhidas três de acordo com o seu tamanho, polidispersividade e eficiência de incorporação, PE (0,6)mM-CWSPs, DODAB:MO (0,1mM)-CWSPs e DODAB:MO (0,3mM)-CWSPs. A citotoxicidade in vitro das formulações selecionadas foi determinada não tendo sido observados valores significativos de citotoxicidade. Com a informação obtida neste trabalho, ambos os sistemas PE-CWSPs e DODAB:MOCWSPs foram considerados apropriados para o desenvolvimento de uma vacina contra as infeções provocadas por C. albicans

Confinement During the COVID-19 Pandemic After Metabolic and Bariatric Surgery—Associations Between Emotional Distress, Energy-Dense Foods, and Body Mass Index

Funding Information: The authors gratefully acknowledge the participants enrolled in the current study for their kindness, generosity, and time in contributing with their personal information. We also acknowledge the support from the Obesity and Metabolic Surgery Unit of Hospital CUF Tejo, CUF Healthcare Group, and the support of the Lifestyle Medicine Unit of CUF Healthcare Group by NOVA Medical School. Publisher Copyright: © 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.Purpose: To estimate the association of emotional distress with both consumption of energy-dense micronutrient-poor foods (EDF) and body mass index (BMI) and the association between EDF consumption and change in BMI, during COVID-19 pandemic in patients with prior bariatric surgery. Materials and Methods: This cross-sectional study applied an online structured questionnaire to 75 postoperative bariatric patients during the first Portuguese lockdown. Emotional distress was assessed trough the Hospital Anxiety and Depression Scale (HADS) and dietary intake was evaluated by Food Frequency Questionnaire (FFQ). Self-reported BMI prior to and at the end of confinement was used to compute BMI change. Pre-surgery BMI was computed from measured height and weight from clinical records. Results: After adjustment for education, sex, time since surgery, pre-surgery BMI, and exercise practice, moderate/severe scores in HADS were significantly positively associated with consumption of EDF (ẞ = 0.799; 95% CI: 0.051, 1.546), but not with BMI. Daily EDF consumption significantly increased the odds of maintaining/increasing BMI (OR = 3.34; 95% CI: 1.18, 9.45), instead of decreasing it (reference). Sweets consumption was the only subcategory of EDF significantly positively associated with the odds of a worse outcome in BMI change (OR = 4.01; 95% CI: 1.13, 14.22). Conclusions: Among postoperative bariatric patients, higher reported levels of emotional distress during confinement are associated with increased EDF consumption. Increased EDF consumption during confinement, particularly sweets, is associated with higher odds of bariatric patients not decreasing their BMI. Additional effort is needed to address inadequate lifestyle behaviors among these patients in the context of the COVID-19 pandemic. Graphical abstract: [Figure not available: see fulltext.]publishersversionpublishe

Genetic variability of Candida albicans SAP8 propeptide in isolates from different types of infection

The secreted aspartic proteases (Saps) are among the most studied virulence determinants in Candida albicans. These proteins are translated as pre-pro-enzymes consisting of a signal sequence followed by a propeptide and the mature enzyme. The propeptides of secreted proteinases are important for the correct processing, folding/secretion of the mature enzyme. In this study, the DNA sequences of C. albicans Saps were screened and a microsatellite was identified in SAP8 propeptide region. The genetic variability of the repetitive region of Sap8 propeptide was determined in 108 C. albicans independent strains isolated from different types of infection: oral infection (OI), oral commensal (OC), vulvovaginal candidiasis (VVC), and bloodstream infections (BSI). Nine different propeptides for Sap8 processing were identified whose frequencies varied with the type of infection. OC strains presented the highest gene diversity while OI isolated the lowest. The contribution of the Saps to mucosal and systemic infections has been demonstrated and recently Sap8 has been implicated in the cleavage of a signalling glycoprotein that leads to Cek1-MAPK pathway activation. This work is the first to identify a variable microsatellite in the propeptide of a secreted aspartic protease and brings new insights into the variability of Sap8.This research was supported by FEDER funds through the Operational Programme COMPETE and national funds through Fundação para a Ciência e Tecnologia (FCT), in the scope of project PEst-OE/BIA/UI4050/2014.info:eu-repo/semantics/publishedVersio

Editorial: Glial plasticity in depression

info:eu-repo/semantics/publishedVersio

Evolução das disponibilidades de alguns alimentos em Portugal de 1916 a 2000

Neste trabalho analisou-se a evolução das disponibilidades de alguns alimentos em Portugal de 1916 a2001, nomeadamente cereais panificáveis (trigo, centeio e milho), arroz, batata, vinho e azeite. Analisámosainda a evolução das disponibilidades de leguminosas secas, mas apenas de 1936 a 2001.As disponibilidades da maioria dos alimentos desceram, nalguns casos substancialmente, de 1916-1925para 1996-2001. As excepções são a batata, o arroz e o trigo, cujas disponibilidades variaram, neste período,+318%, +100% e +59%, respectivamente.As disponibilidades dos restantes alimentos descresceram. Entre 1916-1925 e 1996-2001, as disponibilidadesde milho variaram em -83%, as de centeio em -74%, as de azeite em -17% e as de vinho em -0,3%.Quanto às leguminosas secas, apenas analisámos dados de 1936-1945 a 1996-2001, tendo-se verificadouma variação de -31%.This analysis studied the trends of some food availability in Portugal between 1916 and 2001, namely breadcereals (wheat, rye and maize), rice, potato, wine and olive oil. Additionally, the availability of legumeswas studied, but only for the period of 1936 till 2001.The availability of most foods decreased between de decade of 1916-1925 and the period of 1996-2001. Exceptionsto this rule were the availabilities of potato, rice and wheat, which increased by +318%, +100%and +59%, respectively.All other food availabilities decreased between 1916-1925 and 1996-2001. Maize's availability decreased (-83%), rye's availability decreased (-74%), as well as olive oil's (-17%) and wine's (0,3%) availabilities. Whereit concerns legumes, we concluded that, for the period of 1936-2001, its availability decreased 31%

Portugal e o padrão alimentar mediterrânico.

Objectivos: Avaliar a relação de proximidade entre as disponibilidades alimentares portuguesas e as de quatro países banhados pelo Mediterrâneo. Avaliar ainda se as capitações diárias estiveram (ou estão) próximas das consideradas típicas da alimentação mediterrânica.Métodos: Comparámos as disponibilidades alimentares de Portugal com as da Grécia, Espanha, Itália e França, no seio da antiga União Europeia dos 15, usando as balanças alimentares da Food and Agricultural Organization e analisando quatro décadas (1961-2000). Comparámos também as capitações diárias portuguesas com as descritas como típicas da alimentação mediterrânica pelo Estudo dos Sete Países e pelo Estudo EURATOM. Discussão: Existiu, aparentemente, um grupo de países (Portugal, Grécia, Itália, Espanha e França) com disponibilidades alimentares semelhantes. Ao comparar as capitações portuguesas com os dois estudos referidos, verificou-se existir uma proximidade entre Portugal e Grécia, bem como entre Portugal e Sul de Itália.Conclusões: Apesar de na década de 60 existir um núcleo de países cujas disponibilidades e capitações alimentares indicam a proximidade de um padrão alimentar mediterrânico, foi-se verificando um distanciamento destes países.Objectives: We proposed to evaluate Portugal's food availabilities compared to the availabilities of four Mediterranean countries (Greece, Spain, Italy and France). Furthermore, we proposed to evaluate if Portugal's food availabilities were (or are) near to the ones described as typical of the Mediterranean Diet.Methods: Data from four decades (1961-2000) of FAO's Food Balance Sheets was analyzed. We also compared Portugal's availabilities with data from two well known studies (Seven Countries and EURATOM studies). Results: Where it concerns the data from FAO's food balance sheets, there was a group of countries (Portugal, Greece, Italy, Spain and France) with similar availabilities. As for the comparison between Portugal and data from the two mentioned studies, we observed an alternated proximity Portugal - Greece / Portugal - South of Italy.Conclusions: In the 60's there was a group of countries with avalabilities close to the ones described for the Mediterranean Diet. This situation changed over time

Production, Processing, and Storage at the Early Medieval Site of Senhora Do Barrocal (Municipality of Sátão, Central Portugal)

UIDB/00749/2020 UIDP/00749/2020Small rural places are largely absent from early medieval written sources, but they were profuse and relevant in regional settlements and economies. Only through archaeological and archaeobotanical investigation is it possible to unveil their structure and productive strategies; however, this kind of investigation is still uncommon in Iberia. Here, the assemblage of fruits/seeds, wood charcoal, and food remains from Senhora do Barrocal (SB) (Sátão, Portugal) will be presented and discussed in order to understand the crop production, processing, and storage. The site was destroyed by a fire somewhere between the 10th and the 11th centuries AD, which allowed the preservation of abundant plant remains in a storage area. Charcoal analyses suggest that the building was made with oak and chestnut timber. The massive fruits/seeds assemblage was dominated by cereals, mostly oat and rye, but also barley, millet, and naked wheat, some fully dehusked, others still hulled. Furthermore, evidence of food products has also been found, suggesting that the area was used for the storage of multiple foods and crops at different processing stages. SB is a good example of how communities adopted a diverse set of crops and multifaceted storage strategies to prevent food shortages and to endure in a harsh environment.publishersversionpublishe

Influence of thickness and coatings morphology in the antimicrobial performance of zinc oxide coatings

"Author's copy"In this research work, the production of undoped and silver (Ag) doped zinc oxide (ZnO) thin films for food-packaging applications were developed. The main goal was to determine the influence of coatings morphology and thickness on the antimicrobial performance of the produced samples. The ZnO based thin films were deposited on PET (Polyethylene terephthalate) substrates by means of DC reactive magnetron sputtering. The thin films were characterized by optical spectroscopy, X-Ray Diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and Scanning Electron Microscopy (SEM). The antimicrobial performance of the undoped and Ag-doped ZnO thin films was also evaluated. The results attained have shown that all the deposited zinc oxide and Ag-doped ZnO coatings present columnar morphology with V-shaped columns. The increase of ZnO coatings thickness until 200 nm increases the active surface area of the columns. The thinner samples (50 and 100 nm) present a less pronounced antibacterial activity than the thickest ones (200–600 nm). Regarding Ag-doped ZnO thin films, it was verified that increasing the silver content decreases the growth rate of Escherichia coli and decreases the amount of bacteria cells present at the end of the experiment.The work described in this paper was supported by project NANOPACKSAFER: NANO-engineered PACKaging systems for improving quality, SAFEty and health characteristics of foods, Portugal-Spain International Nanotechnology Laboratory Nanotechnology Projects Call; and also by the FEDER funding through the COMPETE program and FCT PEst-C/BIA/UI4050/2011 project