Application of a robust analytical method for quantifying progestins in environmental samples from three Portuguese Estuaries.

In the last years, progestins have raised special concerns for their documented negative effects on aquatic species, yet little is known about their environmental levels in surface waters and bioaccumulation in the trophic web. This study aimed to 1) adapt an extraction method for quantifying progestins in freeze-dried matrices, 2) validate the analytical procedure for three matrices: bivalve, polychaete, and crustacean, and 3) characterize levels of the four most prescribed synthetic progestins in key species across three Portuguese estuaries. Through the validated method, progestins were only quantifiable for the crustacean. Values were generally low, peaking with drospirenone values in Ria de Aveiro (1.33 ± 0.26 ng/g ww) and Tagus estuary (1.42 ± 0.55 ng/g ww), while Ria Formosa exhibited the lowest progestin concentrations (< 1 ng/g ww). This study enabled the development of a precise extraction and analytical method for quantifying steroid hormones in three distinct biological matrices.

Baseline progestins characterization in surface waters of three main Portuguese estuaries.

Synthetic progestins are micropollutants of special concern, due to their growing use in human and veterinary therapies and their risks to aquatic life. Currently, there is a lack of environmental information on these compounds, worldwide. The main objective of this work was to characterize the levels of the most consumed progestins in Portugal. For that, Ria de Aveiro, Tagus estuary and Ria Formosa were sampled in a temporal perspective to evaluate levels of drospirenone (DRO), desogestrel (DSG), gestodene (GST) and levonorgestrel (LNG). Drospirenone and desogestrel were the most abundant progestins. In the North of Portugal, DSG was the most abundant (Aveiro: 193.9 ng L-1 in summer), while DRO was more representative in the South (Tagus: 178.9 ng L-1; Formosa: 125.7 ng L-1) and also in summer. These spatial differences can be associated with the hydrodynamics of each estuarine system as well as the distinct population and tourist levels associated with each site.

Non-convulsive status epilepticus as the initial manifestation in a family with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an autosomal dominant small-vessel disease caused by mutations of the NOTCH3 gene. It typically presents with migraine, recurrent brain ischaemia, and cognitive disorders. Seizures rarely present as the initial manifestation, with non-convulsive status epilepticus being even less frequent. We present a series of 3 related patients with this arteriopathy, 2 of whom presented status epilepticus as a manifestation of the disease.

Nonlinear Thomson scattering with ponderomotive control.

In nonlinear Thomson scattering, a relativistic electron reradiates the photons of a laser pulse, converting optical light to x rays or beyond. While this extreme frequency conversion offers a promising source for probing high-energy-density materials and driving uncharted regimes of nonlinear quantum electrodynamics, conventional nonlinear Thomson scattering has inherent trade-offs in its scaling with laser intensity. Here we discover that the ponderomotive control afforded by spatiotemporal pulse shaping enables regimes of nonlinear Thomson scattering that substantially enhance the scaling of the radiated power, emission angle, and frequency with laser intensity. By appropriately setting the velocity of the intensity peak, a spatiotemporally shaped pulse can increase the power radiated by orders of magnitude. The enhanced scaling with laser intensity allows for operation at significantly lower electron energies or intensities.

Influence of oceanic and climate conditions on the early life history of European seabass Dicentrarchus labrax.

Understanding how marine fish early-life history is affected in the long-term by environmental and oceanographic factors is fundamental given its importance to population dynamics and connectivity. This work aimed at determining the influence of these processes on the interannual variability in hatch day and early-life growth patterns of European seabass, over a seven-year period (2011-2017) in the Atlantic Iberian coast. To accomplish this, otolith microstructure analysis was used to determine seabass hatch day and to develop early-growth correlations. In most years, hatching occurred from February to April, with two exceptions: in 2012, hatching started in early-January, and in 2016 an exceptionally long hatching period was registered. Using generalized additive models (GAM), we observed that sea surface temperature (SST), the North Atlantic Oscillation index (NAOi) and Chlorophyll-a (Chla) were the main drivers behind the inter-annual variability in seabass hatch day. Analysis of correlations between growth increments allowed assessing important periods of seabass growth and how future growth is affected. Since seawater temperature is among the main drivers for seabass recruitment and growth, its life cycle may be hampered due to ocean warming and an increasingly unstable climate, with consequences for the natural marine stocks and their harvest.

Non-convulsive status epilepticus as the initial manifestation in a family with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). / Status epilepticus no convulsivo como manifestación inicial en una familia con arteriopatía autosómica dominante cerebral con infartos subcorticales y leucoencefalopatía (CADASIL).

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an autosomal dominant small-vessel disease caused by mutations of the NOTCH3 gene. It typically presents with migraine, recurrent brain ischaemia, and cognitive disorders. Seizures rarely present as the initial manifestation, with non-convulsive status epilepticus being even less frequent. We present a series of 3 related patients with this arteriopathy, 2 of whom presented status epilepticus as a manifestation of the disease.

Signo del tridente en la neurosarcoidosis medular / Trident sign in spinal cord neurosarcoidosis

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[Trident sign in spinal cord neurosarcoidosis]. / Signo del tridente en la neurosarcoidosis medular.

TITLE: Signo del tridente en la neurosarcoidosis medular.

Advances on assessing nanotoxicity in marine fish - the pros and cons of combining an ex vivo approach and histopathological analysis in gills.

The need to overcome logistic and ethical limitations of in vivo nanotoxicity evaluation in marine organisms is essential, mostly when dealing with fish. It is well established that medium/solvent conditions affect dispersion and agglomeration of nanoparticles (NPs), which represents a constraint towards a solid and realistic toxicity appraisal. In this way the pros and cons of an ex vivo approach, using a simplified exposure medium (seawater) and addressing gills histopathology, were explored. The nanotoxic potential of environmentally realistic concentrations of titanium dioxide NPs (TiO2 NPs) was also assessed, disclosing the morpho-functional effects on the gills and the possible uptake/elimination processes. Excised gills of the Senegalese sole (Solea senegalensis) were directly exposed in artificial seawater to 20 and 200 µg L-1 TiO2 NPs, for 2 h and 4 h. Semi-quantitative and quantitative histological analyses were applied. The normal morphology of the gill's epithelia was only slightly altered in the control, reflecting protective mechanisms against the artificiality of the experimental conditions, which, together with the absence of differences in the global histopathological index (Ih), corroborated that the gill's morpho-functional features were not compromised, thereby validating the proposed ex vivo approach. TiO2 NPs induced moderate severity and dissemination of histopathological lesions. After 2 h, a series of compensatory mechanisms occurred in NP treatments, implying an efficient response of the innate defense system (increasing number of goblet cells) and effective osmoregulatory ability (chloride cells proliferation). After 4 h, gills revealed signs of recovery (normalization of the number of chloride and goblet cells; similar Ih), highlighting the tissue viability and effective elimination and/or neutralization of NPs. The uptake of the TiO2 NPs seemed to be favored by the higher particle sizes. Overall, the proposed approach emerged as a high-throughput, reliable, accurate and ethically commendable methodology for nanotoxicity assessment in marine fish.

A macroalgae-based biotechnology for water remediation: Simultaneous removal of Cd, Pb and Hg by living Ulva lactuca.

Metal uptake from contaminated waters by living Ulva lactuca was studied during 6 days, under different relevant contamination scenarios. In mono-metallic solutions, with concentrations ranging from 10 to 100 µg L-1 for Hg, 10-200 µg L-1 for Cd, and 50-1000 µg L-1 for Pb, macroalgae (500 mg L-1, d.w.) were able to remove, in most cases 93-99% of metal, allowing to achieve water quality criteria regarding both surface and drinking waters. In multi-metallic solutions, comprising simultaneously the three metals, living macroalgae still performed well, with Hg removal (c.a. 99%) not being significantly affected by the presence of Cd and Pb, even when those metals were in higher concentrations. Removal efficiencies for Cd and Pb varied between 57 and 96%, and 34-97%, respectively, revealing an affinity of U. lactuca toward metals: Hg > Cd > Pb. Chemical quantification in macroalgae, after bioaccumulation assays demonstrated that all Cd and Hg removed from solution was really bound in macroalgae biomass, while only half of Pb showed to be sorbed on the biomass. Overall, U. lactuca accumulated up to 209 µg g-1 of Hg, up to 347 µg g-1 of Cd and up to 1641 µg g-1 of Pb, which correspond to bioconcentration factors ranging from 500 to 2200, in a dose-dependent accumulation. Pseudo-first order, pseudo-second order and Elovich models showed a good performance in describing the kinetics of bioaccumulation, in the whole period of time. In the range of experimental conditions used, no mortality was observed and U. lactuca relative growth rate was not significantly affected by the presence of metals. Results represent an important contribution for developing a macroalgae-based biotechnology, applied for contaminated saline water remediation, more "green" and cost-effective than conventional treatment methods.

Comparative study on metal biosorption by two macroalgae in saline waters: single and ternary systems.

The biosorption capability of two marine macroalgae (green Ulva lactuca and brown Fucus vesiculosus) was evaluated in the removal of toxic metals (Hg, Cd and Pb) from saline waters, under realistic conditions. Results showed that, independently of the contamination scenario tested, both macroalgae have a remarkable capacity to biosorb Hg and Pb. In single-contaminant systems, by using only c.a. 500 mg of non-pre-treated algae biomass (size <200 µm) per litter, it was possible to achieve removal efficiencies between 96 and 99 % for Hg and up to 86 % for Pb. Despite the higher removal of Hg, equilibrium was reached more quickly for Pb (after 8 h). In multi-contaminant systems, macroalgae exhibited a similar selectivity toward the target metals: Hg > Pb> > Cd, although Pb removal by U. lactuca was more inhibited than that achieved by F. vesiculosus. Under the experimental conditions used, none of the macroalgae was effective to remove Cd (maximum removal of 20 %). In all cases, the kinetics of biosorption was mathematically described with success. Globally, it became clear that the studied macroalgae may be part of simple, efficient, and cost-effective water treatment technologies. Nevertheless, Fucus vesiculosus has greater potential, since it always presented higher initial sorption rates and higher removal efficiencies.

Impairment of mitochondrial energy metabolism of two marine fish by in vitro mercuric chloride exposure.

The goal of this work was to understand the extent of mercury toxic effects in liver metabolism under an episode of acute contamination. Hence, the effects of in vitro mercuric chloride in liver mitochondria were assessed in two commercial marine fish: Senegalese sole (Solea senegalensis) and gilthead seabream (Sparus aurata). Liver mitochondria were exposed to 0.2mgL(-1) of mercury, the average concentration found in fish inhabiting contaminated areas. Mercuric chloride depressed mitochondrial respiration state 3 and the maximal oxygen consumption in the presence of FCCP indicating inhibitory effects on the oxidative phosphorylation and on the electron transport chain, respectively. The inhibition of F1Fo-ATPase and succinate-dehydrogenase activities also corroborated the ability of mercury to inhibit ADP phosphorylation and the electron transport chain. This study brings new understanding on the mercury levels able to impair fish mitochondrial function, reinforcing the need for further assessing bioenergetics as a proxy for fish health status.

Long-term functional changes in an estuarine fish assemblage.

The functional diversity of the fish assemblages of the Mondego estuary was studied for a discontinuous 30-year period (1988-2012). During this time, hydrological changes occurred due to man-induced alterations and weather extremes. These changes led to alterations in the structure and function of the fish community. Species richness and functional richness decreased over time and the fish community started to explore new micro-habitats and food resources. Before severe hydrological changes, the community was dominated by pelagic, detritivorous and species with wider salinity ranges. After, the community became dominated by demersal, benthic, piscivorous and marine species. During a drought, omnivorous became increasingly important, reflecting greater possibilities of using available feeding resources. We have also found an increase in sub-tropical species throughout the years, which might be related to gradual temperature increases at a global scale. This study also confirmed estuaries as extremely important for restocking several commercial species.

Field transplantation of the bivalve Scrobicularia plana along a mercury gradient in Ria de Aveiro (Portugal): uptake and depuration kinetics.

The bioaccumulation and depuration capabilities of mercury by the edible bivalve Scrobicularia plana was studied in a coastal lagoon (Ria de Aveiro, Portugal) through a transplantation experiment. Little information on this topic is available in the literature, especially concerning different tissues' responses to contaminant exposure, but the present study is one of the few works that can surpass this knowledge gap. Organisms from a reference area were transplanted to two different contaminated areas in the Ria de Aveiro. In both areas, the bivalves (i.e., entire organism, digestive gland and the rest of the organism) presented a similar saturation model of mercury accumulation, the digestive gland being the tissue that reached the highest concentrations after 25 days of exposure to the contaminant. During this short uptake period, the transplanted organisms reached 20-30% of the concentrations observed in resident contaminated organisms. After the exposure period, the organisms were transplanted to a clean area for more than 25 days of depuration. At the end of the transplantation period, organisms lost approximately 50% of their mercury body burden (60%: the entire organism and digestive gland; 35%: gills and 40%: the rest of the organism) and the ones from the least contaminated site almost reached the concentrations recorded in the reference area. So, the results suggest that S. plana is a promising biomonitoring species, since it accumulates the contaminant in a considerable extent quite rapidly and at the same time it has a low metal retention capacity (low biological half-life) when exposed to clean sediments.

Recovery trends of Scrobicularia plana populations after restoration measures, affected by extreme climate events.

The Mondego estuary (Portugal) went through different ecological scenarios over the last decades. An eutrophication process led to a decline in the ecosystem quality. The ensuing restoration plan resulted into a gradual ecological recovery, which was impaired by the occurrence of successive extreme climate events that affected dynamics and productivity of key species. In this study we assess the response of the bivalve Scrobicularia plana to the impacts of these events in a recovery scenario, by comparing populations in two different intertidal habitats: a seagrass bed and a sandflat area. As a general tendency, S. plana, which was negatively affected by eutrophication, responded positively to restoration. However, the occurrence of extreme climate events seemed to affect recruitment success, biomass and production, impairing the recovery process. In the seagrass bed, S. plana maintained a stable and structured population, while in the sandflat area recovery clearly reverted into a decline, mainly concerning biomass and production values. This sequence of multiple stressors might have reduced S. plana resilience to further impacts and therefore, understanding the behavior of biological populations following restoration initiatives requires acknowledgement that some changes may not be easily reversible.

Uptake and depuration of PCB-153 in edible shrimp Palaemonetes varians and human health risk assessment.

A medium-term mesocosm exposure study was conducted to elucidate bioaccumulation and depuration of polychlorinated biphenyl congener 153 (PCB-153) in edible shrimp Palaemonetes varians. Over the 15-day exposure period, shrimp under different exposure concentrations exhibited a significant increase in PCB-153 concentration compared with control organisms. Distinct bioaccumulation patterns and uptake rates were observed depending on the exposure concentrations. For low PCB-153 exposure levels (0.25µgL(-1)), accumulation followed a saturation model, reaching an apparent steady state after fifteen days exposure. For intermediate (2.5µgL(-1)) and high PCB-153 levels (25µgL(-1)), accumulation was faster and linear. In addition, the bioaccumulation rate was not proportional to PCB-153 concentration, and the bioaccumulation was higher at intermediate exposure concentrations. Regarding the depuration phase, P. varians lost up to 30% of PCB-153 after 72h and levels continued slowly to decrease until the end of the 30-d experimental period. However, PCB-153 levels in shrimp did not reach background values, and those exposed to moderate and high PCB-153 concentrations presented contamination levels much higher than the regulatory limit for human food consumption (75ngg(-1) ww for Σ6 PCB).

Kinetics of the PO4-P adsorption onto soils and sediments from the Mondego estuary (Portugal).

In order to assess the status of a North Atlantic Ocean Ecoregion (Mondego system, Portugal) after the implementation of a management programme, a study on PO4-P sorption was carried out. Considering that sorption velocity markedly determines the systems promptness to phosphorus external loading, the kinetics of the sorption of PO4-P onto estuarine sediments and the upstream agricultural soils was studied. The pseudo-second order kinetic equation gave an appropriate description of PO4-P sorption onto these soils and sediments, allowing for a quantitative comparison. For soils, the kinetic constants (k2*, µg(-1) g h(-1)) ranged between 0.007 and 0.017 µg(-1) g h(-1) while, in the case of sediments these ranged between 0.008 and 0.012 µg(-1) g h(-1). Results showed that the reduction of water residence time after the management action was relevant for the system status, affecting its reaction capacity to phosphorus loading.

Mercury bioaccumulation and the population dynamics of Mesopodopsis slabberi (Crustacea: Mysidacea) along a mercury contamination gradient.

The mercury bioaccumulation and population dynamics of the mysid Mesopodopsis slabberi was assessed along a mercury gradient in Ria de Aveiro (Portugal). M. slabberi is one of the most important mysid species in European temperate coastal shallow waters playing a key ecological role. Nevertheless, no references were found concerning the possible consequences of the Hg on the trophodynamics of these coastal ecosystems. M. slabberi showed a clear bioaccumulation along the Hg gradient and through life, with mature females reaching the highest concentrations. In terms of population structure, higher densities and biomasses of M. slabberi were assessed in the most contaminated areas contrarily to the least polluted areas. Despite the mercury accumulation in its tissues no strong negative effects on the structure and population dynamics of the species were observed. However, mysids might be important in the transfer of metals from the sediments and zooplankton to higher trophic levels such as fishes, most of them with commercial interest.

Influence of age, sex and breeding status on mercury accumulation patterns in the wandering albatross Diomedea exulans.

Although mercury bio-amplifies through the food chain and accumulates in top predators, mercury concentrations in tissues of the wandering albatross are greater than in any other vertebrate, including closely related species. In order to explore the alternative explanations for this pattern, we measured total mercury concentrations in feathers, plasma and blood cells of wandering albatrosses of known age, sex and breeding status sampled at South Georgia. Mercury concentrations were low in feathers and blood components of chicks, and higher in the feathers of young pre-breeders than in feathers or blood of older pre-breeders and breeding adults. There was no effect of sex on mercury concentrations in the feathers of pre-breeders or breeding adults, whereas levels were significantly higher in blood cells of breeding females than males. The high feather mercury concentrations of young pre-breeders compared with older birds suggest an increase in moult frequency as birds approach maturity.

Organochlorine accumulation on a highly consumed bivalve (Scrobicularia plana) and its main implications for human health.

Contamination by polychlorinated biphenyls (PCBs) and hexachlorobenzene (HCB) was investigated along a spatial gradient in water, sediments and in commercially important bivalve species Scrobicularia plana, from Ria de Aveiro (Portugal). Organochlorines dissolved in water were below detection limit and concerning suspended particulate matter, only PCBs were quantified, ranging from 3.8 to 5.8 ng∙g(-1) DW (Σ13PCBs). There was a distinct spatial gradient regarding PCB accumulation in sediments. The highest concentrations were found in deeper layers and closest to the pollution source, decreasing gradually along a 3 km area. Contamination in sediments exceeded the Canadian and Norwegian sediment quality guidelines, inducing potential toxic effects in related biota. PCBs tended to bioaccumulate throughout S. plana lifespan but with different annual rates along the spatial gradient. The maximum values were found in older individuals up to 3+ years old, reaching 19.4 ng∙g(-1) DW. HCB concentrations were residual and no bioaccumulation pattern was evident. Congeners 138, 153 and 180 were the most accumulated due to their abundance and long-term persistence in the environment. In the inner area of the Laranjo Bay (0.6 km(2)), the species was able to remove up to 0.4 g of PCBs annually from sediments into their own tissues, which is consequently free for trophic transfer (biomagnification). Concerning human health, and despite the high concentrations found in sediments, PCB levels in bivalves do not exceed the limit established by the European Union for fishery products and are largely below tolerable daily intake. Although PCBs in Scrobicularia plana are present at low levels, their impact to human health after consumption over many years might be harmful and should be monitored in future studies.