Browsing by Author "Marques, Ana"
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- DNA damage and oxidative stress responses of mussels Mytilus galloprovincialis to paralytic shellfish toxins under warming and acidification conditions - Elucidation on the organ-specificityPublication . Braga, Ana C.; Pereira, Vitoria; Marcal, Raquel; Marques, Ana; Guilherme, Sofia; Reis Costa, Pedro; Pacheco, MarioCommonly affected by changes in climate and environmental conditions, coastal areas are very dynamic environments where shellfish play an important ecological role. In this study, the oxidative stress and genotoxic responses of mussels (Mytilus galloprovincialis) exposed to paralytic shellfish toxin (PST) - producing dinoflagellates Gymnodinium catenatum were evaluated under i) current conditions (CC: 19 degrees C; pH 8.0), warming (W: 24 degrees C; pH 8.0), acidification (A:19 degrees C; pH 7.6) and iv) combined effect of warming and acidification (WA: 24 degrees C; pH 7.6). Mussels were fed with G. catenatum for 5 days, and to a non-toxic diet during the following 10 days. A battery of oxidative stress biomarkers and comet assay was performed at the peak of toxin accumulation and at the end of the post-exposure phase. Under CC, gills and hepatopancreas displayed different responses/vulnerabilities and mechanisms to cope with PST. While gills presented a tendency for lipid peroxidation (LPO) and genetic damage (expressed by the Genetic Damage Indicator - GDI), hepatopancreas seems to better cope with the toxins, as no LPO was observed. However, the mechanisms involved in hepatopancreas protection were not enough to maintain DNA integrity. The absence of LPO, and the antioxidant system low responsiveness, suggests DNA damage was not oxidative. When exposed to toxic algae under W, toxin-modulated antioxidant responses were observed in both gills and hepatopancreas. Simultaneous exposure to the stressors highlighted gills susceptibility with a synergistic interaction increasing DNA damage. Exposure to toxic algae under A led to genotoxicity potentiation in both organs. The combined effect of WA did not cause relevant interactions in gills antioxidant responses, but stressors interactions impacted LPO and GDI. Antioxidant responses and LPO pointed out to be modulated by the environmental conditions in hepatopancreas, while GDI results support the dominance of toxin-triggered process. Overall, these results reveal that simultaneous exposure to warming, acidification and PSTs impairs mussel DNA integrity, compromising the genetic information due to the synergetic effects. Finally, this study highlights the increasing ecological risk of harmful algal blooms to Mytilus ganoprovincianis populations.
- Evaluation of gold complexes to address bacterial resistance, quorum sensing, biofilm formation, and their antiviral properties against BacteriophagesPublication . Marques, Ana; Carabineiro, Sónia A. C.; Aureliano, M.; Faleiro, LThe worldwide increase in antibiotic resistance poses a significant challenge, and researchers are diligently seeking new drugs to combat infections and prevent bacterial pathogens from developing resistance. Gold (I and III) complexes are suitable for this purpose. In this study, we tested four gold (I and III) complexes, (1) chlorotrimethylphosphine gold(I); (2) chlorotriphenylphosphine gold(I); (3) dichloro(2-pyridinecarboxylate) gold (III); and (4) 1,3-bis(2,6-diisopropylphenyl)imidazole2-ylidene gold(I) chloride, for their antibacterial, antibiofilm, antiviral, and anti-quorum sensing activities. Results reveal that 1 significantly inhibits Escherichia coli DSM 1077 and Staphylococcus aureus ATCC 6538, while 2, 3, and 4 only inhibit S. aureus ATCC 6538. The minimum inhibitory concentration (MIC) of 1 for S. aureus ATCC 6538 is 0.59 µg/mL (1.91 µM), and for methicillin-resistant S. aureus strains MRSA 12 and MRSA 15, it is 1.16 µg/mL (3.75 µM). For E. coli DSM 1077 (Gram-negative), the MIC is 4.63 µg/mL (15 µM), and for multi-resistant E. coli I731940778-1, it is 9.25 µg/mL (30 µM). Complex 1 also disrupts biofilm formation in E. coli and S. aureus after 6 h or 24 h exposure. Moreover, 1 and 2 inhibit the replication of two enterobacteria phages. Anti-quorum sensing potential still requires further clarification. These findings highlight the potential of gold complexes as effective agents to combat bacterial and viral infections.
- Invasive clams (Ruditapes philippinarum) are better equipped to deal with harmful algal blooms toxins than native species (R. decussatus): evidence of species-specific toxicokinetics and DNA vulnerabilityPublication . Braga, Ana C.; Marcal, Raquel; Marques, Ana; Guilherme, Sofia; Vilarino, Oscar; Martins, J. Manuel Leao; Gago-Martinez, Ana; Reis Costa, Pedro; Pacheco, MarioThis study aims to assess and compare the kinetics (accumulation/elimination) of the marine biotoxins okadaic acid (OA) and dinophysistoxin-1 (DTX1 ), between native (Ruditapes decussates) and invasive (Ruditapes philippinarum) clam species, and their genotoxic effects and DNA recover capacity after, exposure to toxic dinoflagellates Promcentrum lima. Clams were fed with P. lima for 5 days and then to non-toxic algae (post-exposure) during other 5 days. Toxin concentrations determined in clams by LC-MS/MS were related with DNA damage and repair assessment through the comet and base excision repair (BER) assays, respectively. Differential accumulation patterns were observed between the invasive and native species. The invasive species consistently and progressively accumulated the toxins during the first 24 h of exposure, while the native clams showed drastic variations in the toxin accumulation. Nevertheless, at the end of a 5 days of exposure period, the native dams presented higher toxin concentrations, nearly reaching the legal regulatory limit for human consumption. In addition, native clams were vastly affected by OA and DTX1, presenting an increment in the DNA damage since the first day, with a correspondent increase in the repair activity. On the other hand, invasive clams were not affected by the dinoflagellate toxins, exhibiting only some signs of the challenge, namely an increase in the DNA repair mechanisms in the post-exposure period. Invasive clams R. philippinanim are better adapted to cope with harmful algal blooms and OA-group toxins than native species. These results may increase farming interest and may lead to new introductions of the invasive clams. In sympatry sites, exposure to OA-group toxins may unbalance dams species biomass and distribution as exposure to toxic dinoflagellates affects the native clams from cellular to a population level, representing a significant threat to development and maintenance of R. decussates populations. (C) 2021 Elsevier B.V. All rights reserved.
- Isolation and characterization of a stress-inducible Dunaliella salina Lcy-β gene encoding a functional lycopene β-cyclasePublication . Ramos, Ana; Coesel, Sacha; Marques, Ana; Rodrigues, Marta; Baumgartner, Alexandra; Noronha, João; Rauter, Amélia; Brenig, Bertram; Varela, J.The halotolerant green alga Dunaliella salina accumulates large amounts of β-carotene when exposed to various stress conditions. Although several studies concerning accumulation and biotechnological production of β-carotene have been published, the molecular basis and regulation of the genes involved in carotenoid biosynthesis in D. salina are still poorly known. In this paper, we report the isolation and regulation of the lycopene β-cyclase (Lcy-β) gene by abiotic stress. The function of this gene was determined by heterologous genetic complementation in E. coli. Gene expression and physiological analyses revealed that D. salina Lcy-β steady-state transcript and carotenoid levels were up-regulated in response to all stress conditions tested (salt, light and nutrient depletion). The results presented here suggest that nutrient availability is a key factor influencing carotenogenesis as well as carotenoid biosynthesis-related gene expression in D. salina.
- Patient-physician discordance in assessment of adherence to inhaled controller medication: a cross-sectional analysis of two cohortsPublication . Jácome, Cristina; Pereira, Ana Margarida; Almeida, Rute; Ferreira-Magalhaes, Manuel; Couto, Mariana; Araujo, Luís; Pereira, Mariana; Correia, Magna Alves; Loureiro, Cláudia Chaves; Catarata, Maria Joana; Maia Santos, Lília; Pereira, João; Ramos, Bárbara; Lopes, Cristina; Mendes, Ana; Cidrais Rodrigues, José Carlos; Oliveira, Georgeta; Aguiar, Ana Paula; Afonso, Ivete; Carvalho, Joana; Arrobas, Ana; Coutinho Costa, José; Dias, Joana; Todo Bom, Ana; Azevedo, João; Ribeiro, Carmelita; Alves, Marta; Leiria Pinto, Paula; Neuparth, Nuno; Palhinha, Ana; Gaspar Marques, João; Pinto, Nicole; Martins, Pedro; Todo Bom, Filipa; Alvarenga Santos, Maria; Gomes Costa, Alberto; Silva Neto, Armandina; Santalha, Marta; Lozoya, Carlos; Santos, Natacha; Silva, Diana; Vasconcelos, Maria João; Taborda-Barata, Luís; Carvalhal, Célia; Teixeira, Maria Fernanda; Alves, Rodrigo Rodrigues; Moreira, Ana Sofia; Sofia Pinto, Cláudia; Morais Silva, Pedro; Alves, Carlos; Câmara, Raquel; Coelho, Didina; Bordalo, Diana; Fernandes, Ricardo M; Ferreira, Rosário; Menezes, Fernando; Gomes, Ricardo; Calix, Maria José; Marques, Ana; Cardoso, João; Emiliano, Madalena; Gerardo, Rita; Nunes, Carlos; Câmara, Rita; Ferreira, José Alberto; Carvalho, Aurora; Freitas, Paulo; Correia, Ricardo; Fonseca, Joao AWe aimed to compare patient's and physician's ratings of inhaled medication adherence and to identify predictors of patient-physician discordance.
- Scale-up and large-scale production of Tetraselmis sp CTP4 (Chlorophyta) for CO2 mitigation: from an agar plate to 100-m(3) industrial photobioreactorsPublication . Pereira, Hugo; Paramo, Jaime; Silva, Joana; Marques, Ana; Barros, Ana; Mauricio, Dinis; Santos, Tamara; Schulze, Peter; Raul J Barros; Gouveia, Luisa; Barreira, Luísa; Varela, J.Industrial production of novel microalgal isolates is key to improving the current portfolio of available strains that are able to grow in large-scale production systems for different biotechnological applications, including carbon mitigation. In this context, Tetraselmis sp. CTP4 was successfully scaled up from an agar plate to 35-and 100-m(3) industrial scale tubular photobioreactors (PBR). Growth was performed semi-continuously for 60 days in the autumn-winter season (17th October -14th December). Optimisation of tubular PBR operations showed that improved productivities were obtained at a culture velocity of 0.65-1.35 m s(-1) and a pH set-point for CO2 injection of 8.0. Highest volumetric (0.08 +/- 0.01 g L-1 d(-1)) and areal (20.3 +/- 3.2 g m(-2) d(-1)) biomass productivities were attained in the 100-m(3) PBR compared to those of the 35-m(3) PBR (0.05 +/- 0.02 g L-1 d(-1) and 13.5 +/- 4.3 g m(-2) d(-1), respectively). Lipid contents were similar in both PBRs (9-10% of ash free dry weight). CO2 sequestration was followed in the 100-m(3) PBR, revealing a mean CO2 mitigation efficiency of 65% and a biomass to carbon ratio of 1.80. Tetraselmis sp. CTP4 is thus a robust candidate for industrial-scale production with promising biomass productivities and photosynthetic efficiencies up to 3.5% of total solar irradiance.
- The preyssler-type polyoxotungstate exhibits anti-quorum sensing, antibiofilm, and antiviral activitiesPublication . Faleiro, Maria Leonor; Marques, Ana; Martins, João; Jordão, Luísa; Nogueira, Isabel; Gumerova, Nadiia I.; Rompel, Annette; Aureliano, M.The increase in bacterial resistance to antibiotics has led researchers to find new compounds or find combinations between different compounds with potential antibacterial action and with the ability to prevent the development of antibiotic resistance. Polyoxotungstates (POTs) are inorganic clusters that may fulfill that need, either individually or in combination with antibiotics. Herein, we report the ability of the polyoxotungstates (POTs) with Wells-Dawson P2W18, P2W17, P2W15, and Preyssler P5W30 type structures to differently affect Gram-negative and Gram-positive microorganisms, either susceptible or resistant to antibiotics. The compound P5W30 showed the highest activity against the majority of the tested bacterial strains in comparison with the other tested POTs (P2W15, P2W17 and P2W18) that did not show inhibition zones for the Gram-negative bacteria, A. baumanii I73775, E. coli DSM 1077, E. coli I73194, K. pneumoniae I7092374, and P. aeruginosa C46281). Generally, the results evidenced that Gram-positive bacteria are more susceptible to the POTs tested. The compound P5W30 was the one most active against S. aureus ATCC 6538 and MRSA16, reaching <0.83 mg·mL−1 (100 µM) and 4.96 mg·mL−1 (600 µM), respectively. Moreover, it was verified by NMR spectroscopy that the most promising POT, P5W30, remains intact under all the experimental conditions, after 24 h at 37 ◦C. This prompted us to further evaluate the anti-quorum sensing activity of P5W30 using the biosensor Chromobacterium violaceum CV026, as well as its antibiofilm activity both individually and in combination with the antibiotic cefoxitin against the methicillin-resistant Staphylococcus aureus 16 (MRSA16). P5W30 showed a synergistic antibacterial effect with the antibiotic cefoxitin and chloramphenicol against MRSA16. Moreover, the antibiofilm activity of P5W30 was more pronounced when used individually, in comparison with the combination with the antibioticcefoxitin. Finally, the antiviral activity of P5W30 was tested using the coliphage Qβ, showing a dosedependent response. The maximum inactivation was observed at 750 µM (6.23 mg·mL−1 ). In sum, P5W30 shows anti-quorum sensing and antibiofilm activities besides being a potent antibacterial agent against S. aureus and to exhibit antiviral activities against enteric viruses.