Valorization of traditional portuguese apples and cherries: biochemical characterization and development of functional ingredients

Dissertation presented to obtain a Ph.D degree in Engineering and Technology Sciences, Biotechnology at the Instituto de Tecnologia Química e Biológica, Universidade Nova de LisboaThe consumption of fruits and vegetables is widely recognized to be beneficial to human health and this is mainly attributed to their high content in functional constituents, such as vitamins, carotenoids and polyphenols. In Portugal, the annual production of apples and cherries is about 300,000 and 17,000 ton, respectively. Between all varieties produced, there are some traditional fruits, which are only cultivated in specific regions of the country. In general, these traditional varieties are very appreciated by consumers due to their unique sensorial properties. However, they are less produced and thus more expensive than exotic cultivars. Within this context, this thesis focused on the valorization of these traditional Portuguese fruits in order to increase their productivity and competition in the market.(...)With financial support from FCT, under contract SRFH/BD/19238/200

Medicina e Cirurgia de Animais de Companhia

Mestrado Integrado em Medicina VeterináriaMaster Degree in Veterinary Medicin

Scalable culture strategies for the expansion of patient-derived cancer stem cell lines

Cancer stem cells (CSCs) have recently raised great interest as a promising biological system for designing effective cancer therapies. The scarcity of CSCs in vivo and the consequent low numbers obtained from biopsies represent a major hurdle to the development of such strategies. It is therefore necessary to design robust scalable methods to enable efficient expansion of bona fide CSCs in vitro. Here, we evaluated the applicability of computer-controlled bioreactors combined with 3D aggregate culture and microcarrier technology, widely used in stem cell bioprocessing, for the expansion and enrichment of CSCs isolated from different types of solid tumors—colorectal cancer (CRC) and non-small-cell lung cancer (NSCLC) from two patients. Results show that these culture strategies improved cell expansion and CSC enrichment. Both patient-derived CSC lines were able to grow on microcarriers, the best results being achieved for PPlus 102-L, Pro-F 102-L, Fact 102-L, and CGEN 102-L beads (5-fold and 40-fold increase in total cell concentration for CRC and NSCLC cells, respectively, in 6 days). As for 3D aggregate culture strategy, the cell proliferation profile was donor dependent. NSCLC cells were the only cells able to form aggregates and proliferate, and the flat-bottom bioreactor vessel equipped with a trapezoid-shaped paddle impeller was the most efficient configuration for cell growth (21-fold increase in cell concentration achieved in 8 days). Serum-free medium promotes CSC enrichment in both 3D aggregate and microcarrier cultures. The protocols developed herein for CSC expansion have the potential to be transferred to clinical and industrial settings, providing key insights to guide bioprocess design towards the production of enriched CSC cultures in higher quantity and improved quality.publishe

Protocol for a Scoping Review Abstract

©Sílvia Rego, Ana Rita Henriques, Sofia Silvério Serra, Teresa Costa, Ana Maria Rodrigues, Francisco Nunes. Originally published in JMIR Research Protocols (https://www.researchprotocols.org), 26.10.2023.BACKGROUND: Clinical trials often use digital technologies to collect data continuously outside the clinic and use the derived digital endpoints as trial endpoints. Digital endpoints are also being developed to support diagnosis, monitoring, or therapeutic interventions in clinical care. However, clinical validation stands as a significant challenge, as there are no specific guidelines orienting the validation of digital endpoints. OBJECTIVE: This paper presents the protocol for a scoping review that aims to map the existing methods for the clinical validation of digital endpoints. METHODS: The scoping review will comprise searches from the electronic literature databases MEDLINE (PubMed), Scopus (including conference proceedings), Embase, IEEE (Institute of Electrical and Electronics Engineers) Xplore, ACM (Association for Computing Machinery) Digital Library, CENTRAL (Cochrane Central Register of Controlled Trials), Web of Science Core Collection (including conference proceedings), and Joanna Briggs Institute Database of Systematic Reviews and Implementation Reports. We will also include various sources of gray literature with search terms related to digital endpoints. The methodology will adhere to the Joanna Briggs Institute Scoping Review and the Guidance for Conducting Systematic Scoping Reviews. RESULTS: A search for reviews on the existing evidence related to this topic was conducted and has shown that no such review was previously undertaken. This review will provide a systematic assessment of the literature on methods for the clinical validation of digital endpoints and highlight any potential need for harmonization or reporting of methods. The results will include the methods for the clinical validation of digital endpoints according to device, digital endpoint, and clinical application goal of digital endpoints. The study started in January 2023 and is expected to end by December 2023, with results to be published in a peer-reviewed journal. CONCLUSIONS: A scoping review of methodologies that validate digital endpoints is necessary. This review will be unique in its breadth since it will comprise digital endpoints collected from several devices and not focus on a specific disease area. The results of our work should help guide researchers in choosing validation methods, identify potential gaps in the literature, or inform the development of novel methods to optimize the clinical validation of digital endpoints. Resolving these gaps is the key to presenting evidence in a consistent way to regulators and other parties and obtaining regulatory acceptance of digital endpoints for patient benefit. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/47119.publishersversionpublishe

Relating Bioactivities with the Phenolic Compounds

Funding Information: V.W. acknowledges the Fundação para a Ciência e Tecnologia for PhD fellowship 2021.08511.BD. This work was supported by the Associate Laboratory for Green Chemistry—LAQV, which is financed by national funds from FCT/MCTES (UIDB/50006/2020). The authors acknowledge the financial support from the Fundação para a Ciência e Tecnologia and Portugal 2020 to the Portuguese Mass Spectrometry Network (LISBOA-01-0145-FEDER-402-022125). This work was funded by the Fundação para a Ciência e Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). A.T.S. also thanks FCT/MCTES for the individual grant CEECIND/04801/2017. This research was also funded by the Sociedade Ponto Verde—Sociedade de Resíduos de Embalagens, S.A., through the project entitled “Bioplastics and Edible, Vegan Films”. Publisher Copyright: © 2023 by the authors.Arabinoxylan has prebiotic properties, as it is able to resist digestion in the small intestine and undergoes fermentation in the large intestine. In this work, arabinoxylan was extracted from corn fiber using an alkaline solution and further purified with membrane processing. It was found that the extracts were mainly composed of xylose (50–52%), arabinose (37–39%), galactose (9%) and glucose (1–4%), with an A/X ratio of 0.72–0.77. All the extracts were composed of phenolic compounds, including ferulic acid derivatives such as dimers, trimers and tetramers. The purified extract had a lower concentration of ferulic and p-coumaric acid (0.004 and 0.02 mg/mgdry_weight, respectively) when compared to raw extract (19.30 and 2.74 mg/mgdry_weight, respectively). The same effect was observed for the antioxidant activity, with purified extracts having a lower value (0.17 ± 0.02 µmol TEAC/mg) when compared to the raw extract (2.20 ± 0.35 µmol TEAC/mg). The purified extract showed a greater antiproliferative effect against the HT29 cell line with EC50 = 0.12 ± 0.02 mg/mL when compared to the raw extract (EC50 = 5.60 ± 1.6 mg/mL). Both raw and purified extracts did not show any cytotoxicity to the Caco-2 cell line in the maximum concentration tested (10 mg/mL).publishersversionpublishe

Effect of a sardine supplement on C-reactive protein in patients receiving hemodialysis

Objective - The study evaluated the effect of a canned sardine supplement in C-reactive protein (CRP) in patients on hemodialysis (HD) and the compliance and adherence to this supplement. Design - This was a quasi-experimental study: Participants with a serum CRP of 5 mg/dL or less volunteered to consume a sardine supplement or were maintained on the usual cheese/ham sandwich supplement. Setting - The study took place in two outpatient dialysis units in Lisbon, Portugal. Patients - The study comprised 63 patients receiving maintenance HD three times per week for at least 6 months and an initial CRP concentration of 5 mg/dL or less. Exclusion criteria included the presence of graft vascular access or history of cancer. Intervention - After a 4-week washout period, the nutritional intervention included a canned sardine sandwich for the case group (n = 31) and a cheese or ham sandwich for the control group (n = 32), to be ingested during each routine HD session, 3 times per week, for 8 weeks. Main outcome measure - Serum levels of high-sensitivity CRP were the outcome measure. Results - Only 65 patients from the invited 186 patients met the inclusion criteria and agreed to eat the sardine sandwich supplement three times per week and were involved in the study. A significant proportion of 48% (n = 31, case group) consumed the sardine sandwich supplement three times per week for 8 weeks, fulfilling the requirements and completing the study. The present investigation showed that a sardine sandwich supplement had no effect on CRP levels among patients on HD. However, when participants were stratified according to tertiles of CRP distribution values at baseline, a reduction in CRP levels was found for those in the higher tertile, being higher for the case group (P = .047). Although diabetic patients were excluded from the analysis (eight in the sardine supplementation group and seven in the control group) a significant CRP reduction was found (P = .034). Conclusion - Although a supplement of low-dose n-3 long-chain polyunsaturated fatty acids had no effect on the plasma high-sensitivity CRP of the supplemented group, a reduction in CRP levels was found when patients were stratified for tertiles of CRP (for the upper tertile) and diabetic status (for nondiabetic patients). These findings need to be further confirmed. This canned sardine supplement was accepted by an important proportion of patients, enhancing diet variety and contributing for a greater n-3 long-chain polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid intake

Variación ontogénica en la dieta y estrategia alimentaria de Raja undulata Lacepède, 1802 (Chondrichthyes: Rajidae) en la plataforma continental portuguesa

Under the scope of an ecosystem approach to fisheries, the understanding of trophic interactions is important for the assessment and consequently the proposal of suitable management measures. Raja undulata, like other rajids, is an important demersal predator in the Portuguese coastal community for which the biological and ecological information is still scarce. The ontogenetic dietary shift was investigated. Major length groups were defined through cluster analysis of the mean abundance of prey items. Prey diversity and feeding strategy were evaluated by length group. Multivariate analysis of variance was performed to test the influence of the factors sex, season and maturity on the diet of this species. It was concluded that R. undulata has a dietary ontogenetic shift within the definition of three major length groups (MLGs): 200-550 mm; 550-750 mm and 750-1000 mm. Diet varied from small and semi-pelagic to large and benthic prey. The feeding strategy of the species also changed from a generalised to a specialised diet. The decapod Polybius henslowi was the main prey item, especially for larger predators. Differences were found between sexes, maturity stages and seasons in each MLG.En el ámbito de la aproximación ecosistémica a las pesquerías, la comprensión de las interacciones tróficas es esencial para llevar a cabo una adecuada evaluación y gestión de los recursos. Raja undulata, al igual que otros rajiformes, es un importante depredador demersal que está presente en las costas portuguesas y sobre el que existe poca información biológica y ecológica. Se estudió la variación ontogenética en la dieta. Para determinar grupos de tallas se ha realizado un análisis de clusters empleando datos de abundancia media de presas. La diversidad de las presas y su estrategia alimentaria fueron evaluadas por grupos de tallas. Para comprobar la influencia de los factores sexo, estación del año y madurez en la dieta de esta especie se llevó a cabo un análisis multivariante de la varianza. Los resultados obtenidos indican que R. undulata presenta variaciones ontogenéticas en la dieta, habiéndose definido tres grupos de tallas (MLG): 200-550 mm; 550-750 mm and 750-1000 mm. La dieta varió desde organismos pequeños semipelágicos a presas bentónicas de mayor tamaño. Asimismo, se observó un cambio en la estrategia alimentaria de la especie, desde una dieta generalizada a otra más especializada. El decápodo Polybius henslowi constituyó, en concreto para los depredadores de mayor tamaño, la presa principal. Se detectaron diferencias por sexo, estado de madurez y estación del año en cada MLG

Are Virulence and Antibiotic Resistance Genes Linked? A Comprehensive Analysis of Bacterial Chromosomes and Plasmids

Although pathogenic bacteria are the targets of antibiotics, these drugs also affect hundreds of commensal or mutualistic species. Moreover, the use of antibiotics is not only restricted to the treatment of infections but is also largely applied in agriculture and in prophylaxis. During this work, we tested the hypothesis that there is a correlation between the number and the genomic location of antibiotic resistance (AR) genes and virulence factor (VF) genes. We performed a comprehensive study of 16,632 reference bacterial genomes in which we identified and counted all orthologues of AR and VF genes in each of the locations: chromosomes, plasmids, or in both locations of the same genome. We found that, on a global scale, no correlation emerges. However, some categories of AR and VF genes co-occur preferentially, and in the mobilome, which supports the hypothesis that some bacterial pathogens are under selective pressure to be resistant to specific antibiotics, a fact that can jeopardize antimicrobial therapy for some human-threatening diseasesinfo:eu-repo/semantics/publishedVersio

Prevalência de alterações posturais em crianças e adolescentes em escolas do Algarve

Diversos factores contribuem para o desenvolvimento de alterações posturais, principalmente em crianças e adolescentes, podendo estar associados a futuros problemas de saúde do sistema neuro-músculo-esquelético se estas não forem detectadas precocemente. Assim sendo, o objectivo deste estudo foi o de verificar a prevalência de alterações posturais em crianças e adolescentes em escolas do ensino básico no Algarve, através da avaliação postural e do teste de Adams. Os resultados revelaram que 80,5% dos indivíduos apresentam um ombro mais elevado, 59,5% apresentam assimetria do triângulo de Tales, 47,6% manifestam hiperlordose lombar e 62,9% revelam desvios laterais. A presença de gibosidade foi verificada em 67,8% dos alunos e um reduzido número de alunos apresentou alterações ao nível dos membros inferiores. A partir dos dados obtidos, cremos tornar-se necessário a implantação de programas de rastreio dos desvios posturais em crianças e adolescentes que frequentem as escolas, de forma a prevenir estas alterações e/ou tratá-las precocemente. Além disso, faz-se necessária a modificação do ambiente escolar, envolvendo alunos, professores, encarregados de educação e toda a comunidade escolar

Effect of the matrix system in delivery and in vitro bioactivity of microencapsulated Oregano essential oil

Available at ScienceDirectThe effect of encapsulating matrix on retention, protection and delivery of Oregano essential oil (EO) was studied. EO was encapsulated in rice starch porous spheres, inulin and gelatine/sucrose capsules by spray drying. Gelatine/sucrose matrix was also dried by freeze drying. Experimental designs were applied to test the effect of bonding agents and solids content for rice starch and drying temperature and solids content for inulin and gelatine/sucrose systems. The ratio of gelatine/sucrose was also tested. EO was identified (confocal laser scanning microscopy and FT-IR) in all tested matrices and the release profiles, antioxidant activity and antimicrobial activity of encapsulates evaluated. Results showed that the three tested materials are able to encapsulate Oregano EO. Higher diffusion coefficients were obtained for starch microcapsules (about 10 13 m2/s) followed by spray-dried gelatine/sucrose systems (about 10 15 m2/s) and inulin microcapsules (about 10 16 m2/s). Gelatine/sucrose microparticles exhibit high antioxidant and antimicrobial activity while inulin and rice starch microencapsulates ensure higher stabilit