Biosystems and Integrative Sciences Institute

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Publications

Unravelling the dermatological potential of the brown seaweed Carpomitra costata

Publication . Susano, Patrícia; Silva, Joana; Alves, Celso; Martins, Alice; Gaspar, Helena; Pinteus, Susete; Mouga, Teresa; Goettert, Márcia Ines; Petrovski, Zeljko; Branco, Luís B.; Pedrosa, Rui

The ever-increasing interest in keeping a young appearance and healthy skin has leveraged the skincare industry. This, coupled together with the increased concern regarding the safety of synthetic products, has boosted the demand for new and safer natural ingredients. Accordingly, the aim of this study was to evaluate the dermatological potential of the brown seaweed Carpomitra costata. The antioxidant, anti-enzymatic, antimicrobial, photoprotective and anti-inflammatory properties of five C. costata fractions (F1–F5) were evaluated. The ethyl acetate fraction (F3) demonstrated the most promising results, with the best ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (EC50 of 140.1 g/mL) and the capacity to reduce reactive oxygen species (ROS) production promoted by UVA and UVB radiation in 3T3 cells, revealing its antioxidant and photoprotective potential. This fraction also exhibited the highest anti-enzymatic capacity, inhibiting the activities of collagenase, elastase and tyrosinase (IC50 of 7.2, 4.8 and 85.9 g/mL, respectively). Moreover, F3 showed anti-inflammatory potential, reducing TNF- and IL-6 release induced by LPS treatment in RAW 264.7 cells. These bioactivities may be related to the presence of phenolic compounds, such as phlorotannins, as demonstrated by NMR analysis. The results highlight the potential of C. costata as a source of bioactive ingredients for further dermatological applications.

2021Journal article

Open access

Marine invasive species for high-value products' exploration: Unveiling the antimicrobial potential of Asparagopsis armata against human pathogens

Publication . Pinteus, Susete; Lemos, Marco F.L.; Simões, Marco; Alves, Celso; Silva, Joana; Gaspar, Helena; Martins, Alice; Rodrigues, Américo; Pedrosa, Rui

Infectious diseases remain one of the leading global causes of death, mainly due to the increasing number of multi-resistant microorganisms. Therefore, novel antimicrobials are continuously needed. Marine organisms have already proven to be a rich source of bioactive compounds which can be used for the development of novel pharmacological drugs. Within these, seaweeds are an important resource still underexplored. Asparagopsis armata is a marine invasive seaweed that has spread along several coastlines of the world, inducing negative pressures in marine ecosystems. Therefore, it is crucial to develop strategies to counteract their impacts. In this work, A. armata was extracted with methanol and dichloromethane (1:1) and fractionated by column chromatography with different polarity solvents, providing 8 fractions (F1-F8). All the fractions were evaluated for their antimicrobial potential against important human pathogens, namely Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis, and Candida albicans. Additionally, their capacity to inhibit biofilms formation and the effects on cytoplasmatic membrane and DNA integrity were also assessed. The most active fractions were further purified and submitted to GC–MS analysis. The results showed that A. armata produces compounds with a high inhibitory activity against S. aureus, P. aeruginosa, and C. albicans growth, possibly mediated by cytoplasmatic membrane disruption and DNA damage. GC–MS analysis suggested that the most active fractions were mainly composed of bromoditerpenes and fatty acids. The attained results point to the relevance of the invasive A. armata as a source of antimicrobial substances with broad-spectrum activity. The use of invasive species to obtain natural bioactive compounds presents a two-folded opportunity - high availability of the biological material for the extraction of bioactive compounds and, through specimen collection, the mitigation of the negative effects caused by invasive species, contributing to ecosystem integrity and sustainability.

2020Journal article

Restricted access

Neuroprotective effect of Luteolin-7-O-Glucoside against 6-OHDA-induced damage in undifferentiated and RA-differentiated SH-SY5Y cells

Publication . Rehfeldt, Stephanie Cristine Hepp; Silva, Joana; Pinteus, Susete; Pedrosa, Rui; Alves, Celso; Laufer, Stefan; Goettert, Márcia Inês

Luteolin is one of the most common flavonoids present in edible plants and its potential benefits to the central nervous system include decrease of microglia activation, neuronal damage and high antioxidant properties. The aim of this research was to evaluate the neuroprotective, antioxidant and anti-inflammatory activities of luteolin-7-O-glucoside (Lut7). Undifferentiated and retinoic acid (RA)-differentiated SH-SY5Y cells were pretreated with Lut7 and incubated with 6-hydroxydopamine (6-OHDA). Cytotoxic and neuroprotective effects were determined by MTT assay. Antioxidant capacity was determined by DPPH, FRAP, and ORAC assays. ROS production, mitochondrial membrane potential (DYm), Caspase–3 activity, acetylcholinesterase inhibition (AChEI) and nuclear damage were also determined in SH-SY5Y cells. TNF- , IL-6 and IL-10 release were evaluated in LPS-induced RAW264.7 cells by ELISA. In undifferentiated SH-SY5Y cells, Lut7 increased cell viability after 24 h, while in RA-differentiated SH-SY5Y cells, Lut7 increased cell viability after 24 and 48 h. Lut7 showed a high antioxidant activity when compared with synthetic antioxidants. In undifferentiated cells, Lut7 prevented mitochondrial membrane depolarization induced by 6-OHDA treatment, decreased Caspase-3 and AChE activity, and inhibited nuclear condensation and fragmentation. In LPS-stimulated RAW264.7 cells, Lut7 treatment reduced TNF- levels and increased IL-10 levels after 3 and 24 h, respectively. In summary, the results suggest that Lut7 has neuroprotective effects, thus, further studies should be considered to validate its pharmacological potential in more complex models, aiming the treatment of neurodegenerative diseases.

2022Journal article

Open access

Metabolic stability and metabolite profiling of emerging synthetic cathinones

Publication . Lopes, Rita P.; Ferro, Raquel A.; Milhazes, Margarida; Figueira, Margarida; Caldeira, Maria João; Antunes, Alexandra M.M.; Gaspar, Helena

Synthetic cathinones constitute the second largest groups of new psychoactive substances (NPS), which are especially popular among adolescents and young adults. Due to their potential toxicity, the recreational use of these NPS constitute a serious worldwide public health problem. However, their fast appearance in the market renders the continuous updating of NPS information highly challenging for forensic authorities. The unavailability of pharmacokinetic data for emerging NPS is critical for forensic and clinical verifications. With the ultimate goal of having a proactive approach towards the NPS issue, high resolution mass spectrometry was used in the current work to assess preliminary pharmacokinetic data for 8 selected cathinones: 4 reported substances (4-CIC, 3-CMC, 4-CMC and 4-MEAP) and 4 previously unreported ones (3-CIC, 4-MDMB, 4-MNEB and 4-MDMP) for which the emergence on the NSP market is expected to be eminent, were also included in this study. Based on the calculation of pharmacokinetic parameters, half-life and intrinsic clearance, 4-CMC and 4-MDMB are low and high clearance compounds, respectively, and all the remaining cathinones included in this study are intermediate clearance compounds. This fact anticipates the key role of metabolites as suitable biomarkers to extend detection windows beyond those provided by the parent cathinones. Reduction of the keto group and hydroxylation on the alkyl chains were the common metabolic pathways identified for all cathinones. However, the relative importance of these metabolic transformations is dependent on the cathinone substituents. The glucuronic acid conjugation to metabolites stemming for keto group reduction constituted the sole Phase II transformation identified. To our knowledge, this study constitutes the first metabolite profiling of the already reported synthetic cathinones 4-CIC, 3-CMC and 4-CMC. Noteworthy is the fact that 3-CMC accounts for almost a quarter of the quantity of powders seized during 2020. The analytical methods developed, and the metabolites characterized, are now available to be included in routine screening methods to attest the consumption of the 8 cathinones studied.

2023Journal article

Open access

Marine endophytic fungi associated with Halopteris scoparia (Linnaeus) Sauvageau as producers of bioactive secondary metabolites with potential dermocosmetic application

Publication . Calado, Maria da Luz; Silva, Joana; Alves, Celso; Susano, Patrícia; Santos, Débora; Alves, Joana; Martins, Alice; Gaspar, Helena; Pedrosa, Rui; Campos, Maria Jorge

Marine fungi and, particularly, endophytic species have been recognised as one of the most prolific sources of structurally new and diverse bioactive secondary metabolites with multiple biotechnological applications. Despite the increasing number of bioprospecting studies, very few have already evaluated the cosmeceutical potential of marine fungal compounds. Thus, this study focused on a frequent seaweed in the Portuguese coast, Halopteris scoparia, to identify the endophytic marine fungi associated with this host, and assess their ability to biosynthesise secondary metabolites with antioxidative, enzymatic inhibitory (hyaluronidase, collagenase, elastase and tyrosinase), anti-inflammatory, photoprotective, and antimicrobial (Cutibacterium acnes, Staphylococcus epidermidis and Malassezia furfur) activities. The results revealed eight fungal taxa included in the Ascomycota, and in the most representative taxonomic classes in marine ecosystems (Eurotiomycetes, Sordariomycetes and Dothideomycetes). These fungi were reported for the first time in Portugal and in association with H. scoparia, as far as it is known. The screening analyses showed that most of these endophytic fungi were producers of compounds with relevant biological activities, though those biosynthesised by Penicillium sect. Exilicaulis and Aspergillus chevalieri proved to be the most promising ones for being further exploited by dermocosmetic industry. The chemical analysis of the crude extract from an isolate of A. chevalieri revealed the presence of two bioactive compounds, echinulin and neoechinulin A, which might explain the high antioxidant and UV photoprotective capacities exhibited by the extract. These noteworthy results emphasised the importance of screening the secondary metabolites produced by these marine endophytic fungal strains for other potential bioactivities, and the relevance of investing more efforts in understanding the ecology of halo/osmotolerant fungi.

2021Journal article

Open access