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- Metabolic Profile of Four Selected Cathinones in Microsome Incubations: Identification of Phase I and II Metabolites by Liquid Chromatography High Resolution Mass SpectrometryPublication . Lopes, Beatriz T.; Caldeira, Maria João; Gaspar, Helena; Antunes, Alexandra M. M.Consumption of synthetic cathinones, the second largest class of new psychoactive substances (NPS) reported worldwide, represents a serious public health risk. One of the biggest challenges created by the rapid spread of NPS on the illegal drug market is the discovery of selective biomarkers for their detection in biological matrices, which is only possible through the study of their metabolic profile. The synthetic cathinones 4′-methyl-N,N-dimethylcathinone (4-MDMC), 4′-methyl-N,N-diethylcathinone (4-MDEC), 4′-chloro-α-pyrrolidinovalerophenone (4Cl-PVP), and 4′-chloroethylcathinone (4-CEC) are NPS recently seized in Europe, and, with the exception of 4-CEC, no metabolism study was reported for these cathinones. With the ultimate goal of overcoming this gap, these cathinones were incubated in vitro in human and rat liver microsomes in the presence of Phase I and II (glucuronidation) co-factors, using α-pyrrolidinovalerophenone (α-PVP) as positive control. The metabolite identification was performed by liquid chromatography coupled to tandem high resolution mass spectrometry (LC-HRMS/MS). This allowed the identification of multiple Phase I and glucuronide metabolites of the selected cathinones. Additionally, a new glucuronide conjugate, derived from the recreational drug α-PVP, was herein identified for the first time. Importantly, we have demonstrated that 4-MDMC and 4-MDEC can act as prodrugs of the controlled substances 4-MMC and 4-MEC, respectively. The metabolites herein identified are expected to play an important role not only by acting as potential selective biomarkers of the intake of the synthetic cathinones selected for this study but also to understand their potential adverse effects and link these causative agents to toxicities, thereby helping in the treatment of non-fatal intoxications.
- Assessment of the Levels of Pollution and of Their Risks by Radioactivity and Trace Metals on Marine Edible Fish and Crustaceans at the Bay of Bengal (Chattogram, Bangladesh)Publication . Biswas, Krishna Prasad; Hossain, Shahadat; Deb, Nipa; Bhuian, A.K.M. Saiful Islam; Gonçalves, Sílvia C.; Hossain, Shahadat; Hossen, Mohammad BelalMarine environmental pollution is a longstanding global problem and has a particular impact on the Bay of Bengal. Effluent from different sources directly enters rivers of the region and eventually flows into the Bay of Bengal. This effluent may contain radioactive materials and trace metals and pose a serious threat to the coastal environment, in addition to aquatic ecosystems. Using gamma spectrometry and atomic absorption spectrometry, a comprehensive study was carried out on the radioactivity (226Ra, 232Th, 40K, and 137Cs) and trace metal (Cd, Pb, Zn, Cu, Ni, Fe, Mn, and Cr) concentrations, respectively, in fish and crustacean species collected from the coastal belt of the Bay of Bengal (Chattogram, Bangladesh). The analysis showed a noticeable increment in the levels of different radioactive pollutants in the marine samples, although the consumption of the studied fish and crustacean species should be considered safe for human health. Anthropogenic radionu-clide (137Cs) was not detected in any sample. Furthermore, the metal concentrations of a small num-ber of trace elements (Pb, Cd, Cr) were found to be higher in most of the samples, which indicates aquatic fauna are subject to pollution. The estimated daily intake (EDI), target hazard quotient (THQ), hazard index (HI), and target cancer risk (TR) were calculated and compared with the per-missible safety limits. It was found that consuming the seafood from the Bay of Bengal may cause adverse health impacts if consumption and/or means of pollution are not controlled.
- EditorialPublication . Novais, Sara C.; Maranhão, Paulo
- Development of a new multiplex-PCR tool for the simultaneous detection of the fish pathogens Vibrio alginolyticus, Vibrio anguillarum, Vibrio harveyi and Edwardsiella tardaPublication . Pinto, Micaela Ferreira; Baptista, Teresa; Afonso, Clélia Correia NevesDisease assessment and management in cultured aquatic animals is a major concern in commercial aquaculture. Disease outbreaks have direct effects on fish production, causing serious economic losses in this industry. This can be overcome by early detection through molecular high sensitivity tools such as PCR (polymerase chain reaction). One of the most critical steps in the study of bacterial fish diseases is the precise identification of the infectious agent. Considering the damage that some bacteria can cause to fish and humans, the development of a rapid detection method for the four target species that demonstrates to be simple, accurate and low cost is an essential step for the prevention and early treatment of these diseases. Edwardsiella tarda (ACC 36.1), Vibrio harveyi (DSM 19023), Vibrio anguillarum (AQV 55.1) and Vibrio alginolyticus (CECT 521) were selected as targets of a multiplex PCR tool. The multiplex PCR reactions were performed in various reaction conditions, including different annealing temperatures (between 49 °C and 55 °C) and changes in the MgCl2 concentration from 2mMto 8 mM. Best results were obtained at 51 °C and MgCl2 concentration from 4mM to 6 mM. Primers were tested using purified DNA from the respective bacterial strains, yielding bands at expected sizes, which can be identified on agarose gels with clearness and without overlapping sizes. Results indicate that this multiplex PCR tool is suitable for the detection of target pathogens and may, in the future, have extended practical application in commercial aquaculture.
- Cleaning interactions of fishes at São Tomé Island, West AfricaPublication . Vasco Rodrigues, Nuno; Fontes, Jorge; Bertoncini, A.This study reports six new types of cleaning interactions among tropical reef fish species at São Tomé Island, and confirms previously reported interactions for this area. Spicara melanurus was recorded as a cleaner for the first time, and Abudefduf saxatilis was confirmed in this role, following reports from the Southwest Atlantic. Individuals belonging to the family Labridae were the most commonly-observed cleaners, involved in over 85% of recorded interactions. Juveniles of Bodianus speciosus played a key role as cleaners, interacting with at least seven client species. In addition, some of the client fishes, including Paranthias furcifer and Acanthurus monroviae, interacted with at least four cleaner fish species. All cleaner fish species reported in this study were facultative; obligate cleaners appear to be absent from this region.
- Cytotoxic effects of Ridolfia segetum (L.) Moris phytoproducts in cancer cellsPublication . Beeby, Ellie; Magalhães, Mariana; Lemos, Marco F. L.; Pires, Isabel M.; Cabral, CéliaEthnopharmacological relevance: The past few years have witnessed an increasing interest in essential oils (EOs) as potential therapeutic agents against a wide variety of pathologies, including cancer. EOs extracted from Ridolfia segetum (L.) Moris (R. segetum) are a clear example of a phytoproduct with therapeutic applications, as it is widely used in traditional medicine due to its antioxidant and anti-inflammatory properties, and these properties were already validated by previous studies. Although, it is well established that inflammation is a key hallmark of cancer, with a key role promoting tumorigenesis, and being chronic inflammation often associated with tumorigenic processes, there are no previous studies regarding the assessment of the antitumoural potential of R. segetum EOs. Aim of the study: The present study intends to be the first to evaluate the antitumoural proprieties of R. segetum EO phytoproducts in cancer cell models. Materials and methods: For this, R. segetum EOs were extracted from plants collected at either flowering (RS_Fl) or fruiting (RS_Fr) stage. The impact on proliferation and viability of treatment with R. segetum EO extracts was assessed using in vitro 2D and 3D models. Results: Both R. segetum EOs presented effective antiproliferative/viability effects, evidence noted by low IC50 values in 2D models, and significant reduction of spheroid size in 3D in vitro models. Mechanistically, treatment with R. segetum EOs was associated with an altered G1 (associated with p21 stabilisation), and subsequent induction of apoptosis. Conclusions: Overall, these results indicate that R. segetum EOs have potential as suitable antitumoural therapeutic agents.
- Sandy beach macrofaunal assemblages as indicators of anthropogenic impacts on coastal dunesPublication . Bessa, Filipa; Cunha, David; Gonçalves, Sílvia; Marques, João CarlosOcean sandy beaches are iconic recreational assets to society and have undergone rapid degradation caused by both natural and anthropogenic pressures. A major effect of urbanisation on biota stems from artificial structures placed in aquatic systems. While the installation of these structures has been widespread, our understanding of how they impact shoreline habitats and fauna is relatively limited. On the sandy Leirosa Beach, on the European Atlantic coast (Portugal), a decade of cumulative impacts was recorded, with disruption of the frontal dune, dune nourishment and the incorporation of geotextiles to reconstruct the dune and maintain it as an artificial structure. Two years after the dune rehabilitation process at Leirosa Beach, seasonal sampling campaigns (from spring 2010 to winter 2011) were carried out to assess the effects of the artificial dune (and the first signs of its eventual disruption) on macrofauna compared to a natural dune on the same beach. During the study period, the rehabilitated foredune maintained the average slope, with similar sediment characteristics (finer sediments) when compared with the natural dune system, with no physical significant differences being appreciated between the two sites. The macrofaunal assemblages were represented especially by crustaceans (amphipods and isopods), with similar mean total density, taxon richness and diversity between sites. Special attention was given to the supralittoral species, and the results revealed that the natural zone harboured a significantly higher density of sandhopper Talitrus saltator with respect to the artificial zone. PERMANOVA results detailed the significant difference that occurred, particularly during the warmer seasons (spring and summer 2010), the recruitment period described for this species along this coast. In fact, SIMPER analysis revealed that T. saltator accounted for 46% of the dissimilarities between the natural and artificial dune sites. The presence of geotextiles extending across the supralittoral zone appeared, on the one hand, to allow for the presence of talitrids, but the application of textiles around the area prevented them from burrowing, thus causing them to avoid the rehabilitated area. The results highlighted the potentially negative effects of this artificial dune system if textile disruption continues, and the usefulness of T. saltator as an effective and reliable ecological indicator for these ecosystems. Thus, evaluating the risks and magnitude of human interventions is a major challenge for sandy beach ecologists, and baseline information is required if we are to better understand how resident macrofaunal species deal with future impact scenarios.
- Mechanisms of bioinvasions by coastal crabs using integrative approaches – A conceptual reviewPublication . Rato, Lénia D.; Crespo, Daniel; Lemos, Marco F. L.Crustaceans are amongst the most reported invaders of coastal habitats, and predatory brachyuran crabs one of the most successful marine invasive groups. They hold high tolerance to abiotic stress and their genetic, life-history, and behavioural adaptation mechanisms prompt their invasive conspicuousness. However, there is a generalized ambiguity on the specific processes that make these bioinvasions successful. No trait-related patterns have yet been found, mostly since traits attributed solely to non-indigenous crab species lack confirmation by comparison with their native counterparts and across each other. Therefore, this review discusses the available literature on fitness and plasticity of brachyuran crabs under global changes, and advances the increasing importance of addressing native versus non-native invasive species comparisons in the annals of brachyura invasion dynamics. Further on, integrative approaches are highlighted and proposed as innovative tools to disentangle trait-related tolerance, overall crab phenotypic plasticity and further adaptation. These indicator tools will advance the knowledge on crabs’ invasive potential, which ultimately affects upcoming population dynamics and ecosystem services, adding value to an effective management of coastal bioinvasions.
- Study of Temporal Variations in Species–Environment Association through an Innovative Multivariate Method: MixSTATICOPublication . González-Narváez, Mariela; Fernández-Gómez, María José; Mendes, Susana; Molina, José-Luis; Ruiz-Barzola, Omar; Galindo-Villardón, PurificaciónThe study of biotic and abiotic factors and their interrelationships is essential in the preservation of sustainable marine ecosystems and for understanding the impact that climate change can have on different species. For instance, phytoplankton are extremely vulnerable to environmental changes and thus studying the factors involved is important for the species’ conservation. This work examines the relationship between phytoplankton and environmental parameters of the eastern equatorial Pacific, known as one of the most biologically rich regions in the world. For this purpose, a new multivariate method called MixSTATICO has been developed, allowing mixed-type data structured in two different groups (environment and species) to be related and measured on a space–time scale. The results obtained show how seasons have an impact on species–environment relations, with the most significant association occurring in November and the weakest during the month of May (change of season). The species Lauderia borealis, Chaetoceros didymus and Gyrodinium sp. were not observed in the coastal profiles during the dry season at most stations, while during the rainy season, the species Dactyliosolen antarcticus, Proboscia alata and Skeletonema costatum were not detected. Using MixSTATICO, species vulnerable to specific geographical locations and environmental variations were identified, making it possible to establish biological indicators for this region.
- LDAShiny: An R Package for Exploratory Review of Scientific Literature Based on a Bayesian Probabilistic Model and Machine Learning ToolsPublication . De la Hoz-M, Javier; Fernández-Gómez, Mª José; Mendes, SusanaIn this paper we propose an open source application called LDAShiny, which provides a graphical user interface to perform a review of scientific literature using the latent Dirichlet allocation algorithm and machine learning tools in an interactive and easy-to-use way. The procedures implemented are based on familiar approaches to modeling topics such as preprocessing, modeling, and postprocessing. The tool can be used by researchers or analysts who are not familiar with the R environment. We demonstrated the application by reviewing the literature published in the last three decades on the species Oreochromis niloticus. In total we reviewed 6196 abstracts of articles recorded in Scopus. LDAShiny allowed us to create the matrix of terms and documents. In the preprocessing phase it went from 530,143 unique terms to 3268. Thus, with the implemented options the number of unique terms was reduced, as well as the computational needs. The results showed that 14 topics were sufficient to describe the corpus of the example used in the demonstration. We also found that the general research topics on this species were related to growth performance, body weight, heavy metals, genetics and water quality, among others.
