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Research Project
Interdisciplinary Centre of Marine and Environmental Research
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Publications
Valorisation of sea urchin (Paracentrotus lividus) gonads through canning
Publication . Camacho, Carolina; Oliveira, Helena; Serrano, Carmo; Delgado, Inês; Coelho, Inês; Pedro, Sónia; Lourenço, Helena; Bandarra, Narcisa M.; Marques, António; Pessoa, M. Fernanda; Gonçalves, Amparo; Nunes, M. Leonor
Fresh sea urchin (Paracentrotus lividus) gonads are a delicacy with short seasonal availability, very often heterogeneous in size and intrinsic characteristics. This study aimed to valorise this resource through the preparation of canned products (with/without Porphyra spp.) and evaluate their physicochemical and sensory quality (3–12 months). Canning contributed to a decrease in protein, K and most carotenoids contents; and a concentration of lipids, ash, Na and Se levels. A simulated 12-month ageing led to decrease the protein and β-carotene contents; and the Na and lutein levels concentration. The macroalgae addition resulted in an orange, darker and less soft product, with higher carbohydrates, Na, Se and carotenoids contents. A 25 g-dose contributes to significant daily intakes of protein (8–9%), EPA+DHA (47–53%), I (35–62%) and Se (30–47%). The products were commercially stable/sterile and had good sensory acceptance. Overall, canning constitutes a strategy to provide a nutritionally balanced product available all year-round.
Elemental composition and in vitro bioaccessibility assessment of holothuroids
Publication . Sales, S.S.; Lourenço, H.M.; Bandarra, N.M.; Cardoso, C.; Brito, P.; Botelho, M.J.; Gonçalves, S.; Coelho, I.; Delgado, I.; Pessoa, M.F.; Félix, P.M.; Afonso, C.
The elemental composition and bioaccessibility of the wild holothuroids Holothuria arguinensis, Holothuria forskali, and Holothuria mammata was studied. Nutritional and toxicological aspects were evaluated. H. mammata was the richest in Mn, Ni, As, Cd, Pb, and Hg; H. forskali had the highest Cu and I contents, 4.12–4.93 mg/kg dw and 28.02–28.26 mg/kg dw, respectively; and Summer H. arguinensis had high Se content, 4.26 ± 0.08 mg/kg dw. Holothuroids as food may be a dietary Se and I source, with Pb as main hazard. In order to meet the Se Recommended Daily Allowance (RDA), 15–18 g of dried H. arguinensis and 18–33 g of dried H. mammata would have to be consumed everyday. For I, 10 g of dried H. arguinensis, 6–11 g of dried H. forskali, and 20–21 g of dried H. mammata everyday would be required to achieve the I Dietary Recommended Intake (DRI). For meeting Cu dietary requirements, consumption frequencies would have to exceed 330 g dw/day in the most favourable case (Winter H. forskali). Regarding other essential elements, quantities would be much higher. Regarding contaminants, a consumption above 20–21 g/day of dried H. mammata would constitute a Pb risk. Overall, holothuroids can substantially contribute to elemental nutritional requirements.
Fucoxanthin Holds Potential to Become a Drug Adjuvant in Breast Cancer Treatment: Evidence from 2D and 3D Cell Cultures
Publication . Malhão, Fernanda; Macedo, Ana Catarina; Costa, Carla; Rocha, Eduardo; Ramos, Alice Abreu
Fucoxanthin (Fx) is a carotenoid derived from marine organisms that exhibits anticancer activities. However, its role as a potential drug adjuvant in breast cancer (BC) treatment is still poorly explored. Firstly, this study investigated the cytotoxic effects of Fx alone and combined with doxorubicin (Dox) and cisplatin (Cis) on a panel of 2D-cultured BC cell lines (MCF7, SKBR3 and MDA-MB-231) and one non-tumoral cell line (MCF12A). Fucoxanthin induced cytotoxicity against all the cell lines and potentiated Dox cytotoxic effects towards the SKBR3 and MDA-MB-231 cells. The combination triggering the highest cytotoxicity (Fx 10 µM + Dox 1 µM in MDA-MB-231) additionally showed significant induction of cell death and genotoxic effects, relative to control. In sequence, the same combination was tested on 3D cultures using a multi-endpoint approach involving bioactivity assays and microscopy techniques. Similar to 2D cultures, the combination of Fx and Dox showed higher cytotoxic effects on 3D cultures compared to the isolated compounds. Furthermore, this combination increased the number of apoptotic cells, decreased cell proliferation, and caused structural and ultrastructural damages on the 3D models. Overall, our findings suggest Fx has potential to become an adjuvant for Dox chemotherapy regimens in BC treatment.
Effects and Mechanisms of Action of Preussin, a Marine Fungal Metabolite, against the Triple-Negative Breast Cancer Cell Line, MDA-MB-231, in 2D and 3D Cultures
Publication . Seabra, Rosária; Malhão, Fernanda; Correia, Alexandra; Costa, Carla; Kijjoa, Anake; Rocha, Eduardo
Triple-negative breast cancer (TNBC) represents an aggressive subtype of breast cancer (BC) with a typically poorer prognosis than other subtypes of BC and limited therapeutic options. Therefore, new drugs would be particularly welcome to help treat TNBC. Preussin, isolated from the marine sponge-associated fungus, Aspergillus candidus, has shown the potential to reduce cell viability and proliferation as well as to induce cell death and cell cycle arrest in 2D cell culture models. However, studies that better mimic the tumors in vivo, such as 3D cell cultures, are needed. Here, we studied the effects of preussin in the MDA-MB-231 cell line, comparing 2D and 3D cell cultures, using ultrastructural analysis and the MTT, BrdU, annexin V-PI, comet (alkaline and FPG modified versions), and wound healing assays. Preussin was found to decrease cell viability, both in 2D and 3D cell cultures, in a dose-dependent manner, impair cell proliferation, and induce cell death, therefore excluding the hypothesis of genotoxic properties. The cellular impacts were reflected by ultrastructural alterations in both cell culture models. Preussin also significantly inhibited the migration of MDA-MB-231 cells. The new data expanded the knowledge on preussin actions while supporting other studies, highlighting its potential as a molecule or scaffold for the development of new anticancer drugs against TNBC.
New Insights in Saccharomyces cerevisiae Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression
Publication . Valério, Elisabete; Barreiros, Sara; Rodrigues, Sara; Turkina, Maria V.; Vasconcelos, Vitor M.; Campos, Alexandre
Microcystins (MCs) are hepatotoxins produced by some cyanobacteria. They are cyclic peptides that inhibit the serine/threonine protein phosphatases (PPs) PP1 and PP2A, especially PP2A. The inhibition of PP2A triggers a series of molecular events, which are responsible for most MC cytotoxic and genotoxic effects on animal cells. It is also known that MCs induce oxidative stress in cells due to the production of reactive oxygen species (ROS). However, a complete characterization of the toxic effects of MCs is still not accomplished. This study aimed to clarify additional molecular mechanisms involved in MC-LR toxicity, using Saccharomyces cerevisiae as eukaryotic model organism. First, a shotgun proteomic analysis of S. cerevisiae VL3 cells response to 1 nM, 10 nM, 100 nM, and 1 µM MC-LR was undertaken and compared to the control (cells not exposed to MC-LR). This analysis revealed a high number of proteins differentially expressed related with gene translation and DNA replication stress; oxidative stress; cell cycle regulation and carbohydrate metabolism. Inference of genotoxic effects of S. cerevisiae VL3 cells exposed to different concentrations of MC-LR were evaluated by analyzing the expression of genes Apn1, Apn2, Rad27, Ntg1, and Ntg2 (from the Base Excision Repair (BER) DNA repair system) using the Real-Time RT-qPCR technique. These genes displayed alterations after exposure to MC-LR, particularly the Apn1/Apn2/Rad27, pointing out effects of MC-LR in the Base Excision Repair system (BER). Overall, this study supports the role of oxidative stress and DNA replication stress as important molecular mechanisms of MC-LR toxicity. Moreover, this study showed that even at low-concentration, MC-LR can induce significant changes in the yeast proteome and in gene expression.
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Funding agency
Fundação para a Ciência e a Tecnologia
Funding programme
6817 - DCRRNI ID
Funding Award Number
UIDB/04423/2020