IDENTIFICATION AND DESCRIPTION OF BUCEPHALUS MINIMUS (DIGENEA: BUCEPHALIDAE) LIFE CYCLE IN PORTUGAL: MORPHOLOGICAL, HISTOPATHOLOGICAL, AND MOLECULAR DATA

The cercaria of Bucephalus minimus infects the digestive gland and gonads of its first intermediate host, the edible cockle. Cerastoderma edule. Light microscopy (LM) and scanning electron microscopy (SEM) of the cercaria showed a tail formed by a central stem, with 2 long contractile arms presenting distinct morphological surfaces. The encysted metacercaria naturally infected the flathead grey mullet, Mugil cephalus. The cysts found in the heart, liver, and spleen were shown to be identical by the internal transcribed spacer (ITS1) sequence and morphological features and were associated with encapsulation, recruitment of cell infiltrates, and presence of melanomacrophages and adipose tissue. To establish the life cycle. we compared the ITS1 sequence in an adult from the known definitive host, Dicentrarchus labrax: encysted metacercariac from the liver, heart, and spleen of M. cephalus; and a cercaria from C. edule. With this comparison, we determined that they had a 100% similarity. Therefore, the ITS1 sequence data clearly indicate that these 3 parasitic stages belong to the same species, i.e., B. minimus

Differential effects of antiepileptic drugs on human bone cells

Antiepileptic drugs (AED) have been associated to in vivo deleterious consequences in bone tissue. The present work aimed to characterize the cellular and molecular effects of five different AED on human osteoclastogenesis and osteblastogenesis. It was observed that the different drugs had the ability to differentially modulate both processes, in a way dependent on the identity and dose of the AED. Shortly, valproic acid stimulated either osteoclastogenesis and osteoblastogenesis, whereas carbamazepine, gabapentin, and lamotrigine revealed an opposite behavior; topiramate elicited a decrease of osteoclast development and an increase in osteoblast differentiation. This is the first report describing the direct effects of different AED on human primary bone cells, which is a very important issue, because these drugs are usually consumed in long-term therapeutics, with acknowledged in vivo effects in bone tissue.info:eu-repo/semantics/publishedVersio

Evidences for a new role of miR-214 in chondrogenesis

miR-214 is known to play a role in mammalian skeletal development through inhibition of osteogenesis and stimulation of osteoclastogenesis, but data regarding other vertebrates, as well as a possible role in chondrogenesis, remain unknown. Here, we show that miR-214 expression is detected in bone and cartilage of zebrafish skeleton, and is downregulated during murine ATDC5 chondrocyte differentiation. Additionally, we observed a conservation of the transcriptional regulation of miR-214 primary transcript Dnm3os in vertebrates, being regulated by Ets1 in ATDC5 chondrogenic cells. Moreover, overexpression of miR-214 in vitro and in vivo mitigated chondrocyte differentiation probably by targeting activating transcription factor 4 (Atf4). Indeed, miR-214 overexpression in vivo hampered cranial cartilage formation of zebrafish and coincided with downregulation of atf4 and of the key chondrogenic players sox9 and col2a1. We show that miR-214 overexpression exerts a negative role in chondrogenesis by impacting on chondrocyte differentiation possibly through conserved mechanisms.Calouste Gulbenkian Foundation (program "Na Fronteira das Ciencias da Vida"); FCT [UID/Multi/04326/2013, PEst-C/MAR/LA0015/2011, SFRH/BD/38607/2007, SFRH/BPD/45034/2008, SFRH/BPD/111289/2015]; European Commission (ERDF-COMPETE) [PEst-C/MAR/LA0015/2011]info:eu-repo/semantics/publishedVersio

A new insight into the influence of habitat on the biochemical properties of three commercial sea cucumber species

This work makes a comparative evaluation of the biochemical profile of three sea commercial cucumber species (Holothuria mammata, H. polii and H. tubulosa) caught from different locations of the Mediterranean Sea (SE Spain). All species had high levels of moisture (from 73.6% in H. mammata to 81.2% in H. tubulosa), crude ash (from 9.61% in H. mammata to 14.7% in H. tubulosa) and protein (3.01% in H. tubulosa to 11.1% in H. mammata). They also had a low fat content, from 0.21% in H. tubulosa to 0.55% in H. mammata. Holothuria polii had intermediate values between the other two species, for all considered variables. All species had adequate protein/lipid ratios (H. mammata, 20:1; H. polii, 23:1; H. tubulosa, 14:1) and low lipid levels, enriched in omega-3 polyunsaturated fatty acids, especially arachidonic acid. The fatty acid profile suggests that H. polii is feeding on sediments more influenced by terrestrial inputs than the remaining species. Holothuria mammata and H. tubulosa are feeding on marine food sources mainly, but also with some terrestrial influence. The most abundant amino acids detected were alanine, arginine, glutamic acid, and glycine. All species had similar contents of essential amino acids (EAA) and ratios of EAA/non-essential amino acids. Holothuria tubulosa had a high content of toxic metals including Cr, Pb and Ni. This work highlights differences in compositional characteristics between different species of the same genus (Holothuria) from different locations.info:eu-repo/semantics/publishedVersio

Antiproliferative Organic Salts Derived from Betulinic Acid: Disclosure of an Ionic Liquid Selective Against Lung and Liver Cancer Cells

In the last few years, we have been witnessing an increasing interest in ionic liquids (ILs) and organic salts, given their potential applications in biological and pharmaceutical sciences. We report the synthesis and in vitro evaluation of novel organic salts combining betulinate, known for its anticancer properties, with antimalarial drugs, primaquine, chloroquine, and mepacrine, and also with the trihexyltetradecylphosphonium ([P6,6,6,14]) cation. The salts were screened for their in vitro activity against tumor lines HepG2 (liver), MG63 (osteosarcoma), T47D (breast), A459 (lung), and RKO (colon), and also on normal human fibroblasts. All betulinates prepared displayed antiproliferative properties, with the trihexyltetradecylphosphonium betulinate standing out for its higher selectivity. This unprecedented disclosure of a betulinic acid (BA)-derived IL with selective antitumor activity constitutes a relevant first step toward development of novel anticancer therapies based on BA-derived IL.info:eu-repo/semantics/publishedVersio

Evolution during Three Ripening Stages of Évora Cheese

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Anti-Hepatocellular Carcinoma (HepG2) activities of Monoterpene Hydroxy Lactones isolated from the Marine Microalga Tisochrysis Lutea

Tisochrysis lutea is a marine haptophyte rich in omega-3 polyunsaturated fatty acids (e.g., docosahexaenoic acid (DHA)) and carotenoids (e.g., fucoxanthin). Because of the nutraceutical applications of these compounds, this microalga is being used in aquaculture to feed oyster and shrimp larvae. In our earlier report, T. lutea organic crude extracts exhibited in vitro cytotoxic activity against human hepatocarcinoma (HepG2) cells. However, so far, the compound(s) accountable for the observed bioactivity have not been identified. Therefore, the aim of this study was to isolate and identify the chemical component(s) responsible for the bioactivity observed. Bioassay-guided fractionation through a combination of silica-gel column chromatography, followed by preparative thin layer chromatography (PTLC), led to the isolation of two diastereomers of a monoterpenoid lactone, namely, loliolide (1) and epi-loliolide (2), isolated for the first time in this species. The structural elucidation of both compounds was carried out by GC-MS and 1D (1H and 13C APT) and 2D (COSY, HMBC, HSQC-ed, and NOESY) NMR analysis. Both compounds significantly reduced the viability of HepG2 cells and were considerably less toxic towards a non-tumoral murine stromal (S17) cell line, although epi-loliolide was found to be more active than loliolide.FCT, Portugal: UID/Multi/04326/2019, UID/MAR/04292/2020 (MARE), UID/Multi/04046/2020 (BioISI) and from the grant of 0055 ALGARED +5E—INTERREG V-A; SFRH/BD/116604/2016; SFRH/BD/105541/2014info:eu-repo/semantics/publishedVersio

Unlocking the in vitro anti-Trypanosoma cruzi activity of halophyte plants from the southern Portugal

Objective: To evaluate the in vitro anti-Trypanosoma cruzi (T. cruzi) activity of organic extracts prepared from halophyte species collected in the southern coast of Portugal (Algarve), and chemically characterize the most active samples.Methods: Acetone, dichloromethane and methanol extracts were prepared from 31 halophyte species and tested in vitro against trypomastigotes and intracellular amastigotes of the Tulahuen strain of T. cruzi. The most active extract was fractionated by preparative HPLC-DAD, affording 11 fractions. The most selective fraction was fully characterized by H-1 NMR.Results: From 94 samples tested, one was active, namely the root dichloromethane extract of Juncus acutus (IC50 < 20 mu g/mL). This extract was fractionated by HPLC, affording 11 fractions, one of them containing only a pure compound (juncunol), and tested for anti-parasitic activity. Fraction 8 (IC50 = 4.1 mu g/mL) was the most active, and was further characterized by H-1 NMR. The major compounds were phenanthrenes, 9,10-dihydrophenanthrenes and benzocoumarins.Conclusion: Our results suggest that the compounds identified in fraction 8 are likely responsible for the observed anti parasitic activity. Further research is in progress aiming to isolate and identify the specific active molecules. To the best of our knowledge, this is the first report on the in vitro anti T. cruzi activity of halophyte species.PROEP/CNPq/FIOCRUZ 401988/2012-0; FCT/CAPES 2358, 2014/201

A colonização portuguesa na região de Cabo Frio (Rio de Janeiro, Brasil) e o desenvolvimento da atividade piscatória

The Cabo Frio region, the object area of this study, is located in the coastal lowlands of the State of Rio de Janeiro, Brazil. The natural conditions, such as the existence of the Araruama lagoonal system and the occurrence of the resurgence phenomenon on the seashore, provided a rich area for exploitation of marine resources, promoting human settlement and the subsequent development of lagoonal and coastal fishing. Despite the activity was accomplished by indigenous people established in the region, in the seventeen century, with the arrival of Portuguese settlers to Cabo Frio, its development was observed. Taking into account the characteristics of the environment in the provision of fish resources, we intend to analyse diachronically the evolution of the fishing practice, through the investigation of historical documentation.A região de Cabo Frio, área objeto deste estudo, está localizada na baixada litorânea do Estado do Rio de Janeiro, Brasil. As existência do sistema lagunar de Araruama e a ocorrência do fenómeno da ressurgência no seu litoral, proporcionaram uma zona favorável à exploração de recursos marinhos, fomentando a fixação humana e o posterior desenvolvimento da pesca lagunar e costeira. Nesse sentido, a atividade apesar de ser praticada pelos povos indígenas estabelecidos na região, no século dezassete, com a efetivação da colonização portuguesa em Cabo Frio observou-se a sua intensificação e desenvolvimento. Tendo em conta as caraterísticas do meio no propiciar de recursos piscícolas, pretende-se analisar diacronicamente a evolução da prática piscatória na região, por meio da investigação de documentação histórica

Pathogens in ornamental waters: a follow up study

Ornamental waters of easy access and populated with animals are quite attractive and can hide threats to human health. Here we evaluated the microbiota of ornamental waters in a Lisbon park. Water and biofilm samples where collected, in 2 lakes (L1-L2) and ornamental fountains (L3-L4) in February/2015. In May/2015 and monthly during a year (starting March/2016) samples from L4 where collected. Microbiota identification was performed as described previouslya. Biofilm assembly was monitored by crystal violet assay and SEMb and antibiotic susceptibility was performed by conventional methods. The results of the first water sampling (Feb/2015) revealed the presence of Enterobactereaceae and non-fermentative oxidase-positive bacteria. Fountains and lakes presented different microbota being the highest diversity found in L1 hosting a duck population. This result suggested the existence of an interplay between animal inhabitants and microbiota which was confirmed by the second sampling of L4 (May/2015). Between the 2 sampling events a fish population was introduced and the microbiota was completely altered with the appearance of a typical fish pathogen (Aeromonas spp). This tendency was also confirmed over 2016. K. pneumoniae and Aeromonas spp., present as planktonic and biofilm organized bacteria in 2015 showed an enhanced ability to assemble biofilms in vitro at 25 °C than at 37 °C. Bacteria recovered from biofilm showed an increased antibiotic resistance compared to planktonic counterparts. The pilot study conducted during 2015 and the follow up study (still in progress) support a periodic control of ornamental water microbiota as simple preventive measure to avoid potential health issues.Fundação para a Ciência e Tecnologia for the grant PEst-OE/CTM/UI0084/2011N/