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In 2017, aquaculture producers of the Albufeira lagoon, Portugal, reported an invasion of tunicates that was disrupting mussel production, particularly the tunicate Styela plicata (Lesueur, 1823). A totally effective eradication method still does not exist, particularly for S. plicata, and the effects of the eradication treatments on bivalves’ performance are also poorly understood. Our study examined the effectiveness of eradication treatments using three laboratory trials and five treatments (air exposure, freshwater immersion, sodium hypochlorite, hypersaline solution and acetic acid) for S. plicata, as well as their effects on survival and growth of blue mussel Mytilus edulis Linnaeus, 1758. While air exposure and freshwater immersion caused a 27% mortality rate in S. plicata, the acetic acid treatment was the most effective in eliminating this species (>90% mortality). However, a 33–40% mortality rate was registered in mussels. Both species were not affected by the hypersaline treatment in the last trial, but the sodium hypochlorite treatment led to a 57% mortality rate in mussels. Differences in mussels’ growth rates were not detected. These trials represent a step forward in responding to the needs of aquaculture producers. However, further studies are needed to investigate the susceptibility of tunicates to treatments according to sexual maturation, as well as to ensure minimum mussel mortality in the most effective treatments, and to better understand the effects on mussel physiological performance in the long-term.

Pinus pinea nuts are commercial relevant Mediterranean edible forest nuts, with an increasing production and market value, whose industrial processing yields a lignocellulosic by-product, the pine nut shells, currently only used for combustion. Little research has been done on pine nut shells that could support a value-added application for this residue. This work studies for the first time the production of oligosaccharides by autohydrosis, and aims at an integrated upgrade within the biorefinery framework. Autohydrolysis was explored in the temperature range between 150 and 230 °C (corresponding to severity factors 2.13–4.63). Oligosaccharides, mainly xylo-oligosaccharides (95% of the total), were the key soluble products, reaching 28.7 g/100 g of xylan of the feedstock at the optimal conditions (log R0 4.01). Other products were monosaccharides and phenolic compounds that reached 7.8 and 4.7 g/L, respectively, under the most severe conditions. The stability of the oligosaccharides at different temperatures (room, 37 °C and 100 °C) and pH (between 1 and 11) grant them significant market potential in the food and pharma sectors. The pre-treated pine nut shells by autohydrolysis presented an improved, although low, enzymatic digestibility (14%), and an improved high-heating value, therefore advising their further valorization by thermochemical pathways.

Transcranial magnetic stimulation (TMS) can be useful for therapeutic purposes for a variety of clinical conditions. Numerous studies have indicated the potential of this non-invasive brain stimulation technique to recover brain function and to study physiological mechanisms. Following this line, the articles contemplated in this Research Topic show that this field of knowledge is rapidly expanding and considerable advances have been made in the last few years. There are clinical protocols already approved for Depression (and anxiety comorbid with major depressive disorder), Obsessive compulsive Disorder (OCD), migraine headache with aura, and smoking cessation treatment but many studies are concentrating their efforts on extending its application to other diseases, e.g., as a treatment adjuvant. In this Research Topic we have the example of using TMS for pain, post-stroke depression, or smoking cessation, but other diseases/injuries of the central nervous system need attention (e.g., tinnitus or the surprising epilepsy). Further, the potential of TMS in health is being explored, in particular regarding memory enhancement or the mapping of motor control regions, which might also have implications for several diseases. published

The authors are thankful to the Department of Science and Technology (DST), New Delhi, for partially funding the SERB grant (No.: SB/EMEQ). Publisher Copyright: Copyright © 2022 Carletti, Cardoso, Lobo-Arteaga, Sales, Juliao, Ferreira, Chainho, Dionísio, Gaudêncio, Afonso, Lourenço, Cancela, Bandarra and Gavaia. Bone metabolic disorders such as osteoporosis are characterized by the loss of mineral from the bone tissue leading to its structural weakening and increased susceptibility to fractures. A growing body of evidence suggests that inflammation and oxidative stress play an important role in the pathophysiological processes involved in the rise of these conditions. As the currently available therapeutic strategies are often characterized by toxic effects associated with their long-term use, natural antioxidants and anti-inflammatory compounds such as polyphenols promise to be a valuable alternative for the prevention and treatment of these disorders. In this scope, the marine environment is becoming an important source of bioactive compounds with potential pharmacological applications. Here, we explored the bioactive potential of three species of holothurians (Echinodermata) and four species of tunicates (Chordata) as sources of antioxidant and anti-inflammatory compounds with a particular focus on polyphenolic substances. Hydroethanolic and aqueous extracts were obtained from animals’ biomass and screened for their content of polyphenols and their antioxidant and anti-inflammatory properties. Hydroethanolic fractions of three species of tunicates displayed high polyphenolic content associated with strong antioxidant potential and anti-inflammatory activity. Extracts were thereafter tested for their capacity to promote bone formation and mineralization by applying an assay that uses the developing operculum of zebrafish (Danio rerio) to assess the osteogenic activity of compounds. The same three hydroethanolic fractions from tunicates were characterized by a strong in vivo osteogenic activity, which positively correlated with their anti-inflammatory potential as measured by COX-2 inhibition. This study highlights the therapeutic potential of polyphenol-rich hydroethanolic extracts obtained from three species of tunicates as a substrate for the development of novel drugs for the treatment of bone disorders correlated to oxidative stress and inflammatory processes. publishersversion published

Neurofibromatosis type 1 (NF1) is associated with GABAergic dysfunction which has been suggested as the underlying cause of cognitive impairments. Previous intervention trials investigated the statins’ effects using cognitive outcome measures. However, available outcome measures have led to inconclusive results and there is a need to identify other options. Here, we aimed at investigating alternative outcome measures in a feasibility trial targeting cortical inhibition mechanisms known to be altered in NF1. We explored the neurochemical and physiological changes elicited by lovastatin, with magnetic resonance spectroscopy and transcranial magnetic stimulation (TMS). Fifteen NF1 adults participated in this randomized, triple-blind, placebo-controlled crossover trial (Clinicaltrials.gov NCT03826940) composed by one baseline and two reassessment visits after lovastatin/placebo intake (60mg/day, 3-days). Motor cortex GABA+ and Glx concentrations were measured using HERMES and PRESS sequences, respectively. Cortical inhibition was investigated by paired-pulse, input-output curve and cortical silent period (CSP) TMS protocols. CSP ratios were significantly increased by lovastatin (relative: p=0.027; absolute: p=0.034) but not by placebo. CSP durations showed a negative correlation with LICI 50ms amplitude ratio. Lovastatin was able to modulate cortical inhibition in NF1, as assessed by TMS CSP ratios, highlighting the potential of this outcome measure to be considered in future large-scale studies.

Day-to-day we have to make choices. Even simple decisions involve complex cognitive processes that are still not clearly understood. When it comes to critical decisions which may lead to impactful and irreversible consequences, the understanding of the decision-making process is highly relevant. Dysfunctional coping responses may lead to stress and ultimately to trauma. In the present project, we aim to understand decision-making in the context of firefighting and to study how neurocognitive control and stress management strategies affect decision-making. We present here a functional brain imaging paradigm coupled with biosensors while firefighters play a decision-making task. In this computerized task, the firefighter faces the dilemma of entering or not in a burning house, with variable risk of house collapsing, variable number of victims in danger and variable probability of the victims saving themselves. We present here the preliminary results from a cohort of 13 firefighters, of which none presented clinically significant symptoms of post-traumatic stress disorder. We found brain regions, including the ventromedial and dorsolateral prefrontal cortices and angular gyrus, showing a parametric pattern of activation during the decision phase, i.e. the lower the risk of collapsing, the higher the neural activity in these areas. This suggests that these regions are processing that risk information and signalling the chances of being successful in the rescue phase. We also found that the power in high frequencies of the pulse rate variability of this cohort of firefighters is higher when they decide to enter for the rescue. It is suggestive that deciding not to enter for rescue causes an increase in arousal, which may be related to the expectation about the victims’ outcome (whether they survived or not). Concerning coping strategies, we found that active coping used in personal context was significantly correlated with the age and the years of experience in our sample of firefighters. We aim to understand if this pattern of brain activity reflects the coping strategies and if, in turn, it is reflected in the physiological signals we measure. Concurrently, we are recruiting firefighters with post-traumatic stress disorder and non-firefighters to perform the same task. We will investigate whether the decision processes in firefighters are different from non-firefighters, and we will address if post-traumatic stress disorder impairs decision-making in this context. Biosignals will also be used in a second phase of the project to inform training systems about the physiological state of the firefighter during virtual reality simulation. This may help developing optimal neurocognitive control and better coping strategies to deal with stress.

The study of the physiological effects underlying brain response to transcranial magnetic stimulation is important to understand its impact on neurorehabilitation. We aim to analyze the impact of a transcranial magnetic stimulation protocol, the continuous theta burst (cTBS), on human neurophysiology, particularly on contralateral motor rhythms. cTBS was applied in 20 subjects over the primary motor cortex. We recorded brain electrical activity pre- and post-cTBS with electroencephalography both at rest and while performing motor tasks, to evaluate changes in brain oscillatory patterns such as mu and beta rhythms. Moreover, we measured motor-evoked potentials before and after cTBS to assess its impact on brain's excitability. On the hemisphere contralateral to the protocol, we did observe a significant increase in mu (p = 0.027) and beta (p = 0.006) rhythms from pre- to post-cTBS, at the beginning of arm elevation. The topology of action planning and motor execution suggests that cTBS produced an inhibitory effect that propagated to the contralateral hemisphere, thereby precluding the expected/desired excitation for therapy purposes. This novel approach provides support for the notion that this protocol induces inhibitory changes in contralateral motor rhythms, by decreasing desynchronization, contradicting the ipsilateral inhibition vs. contralateral disinhibition hypothesis. Our results have implications for personalized cTBS usage as a rehabilitation intervention, suggesting that an unexpected propagation of inhibition can occur.

Thermography is a non-radiating and contact-free technology which can be used to monitor skin temperature. The efficiency and safety of thermography technology make it a useful tool for detecting and locating thermal changes in skin surface, characterized by increases or decreases in temperature. This work intends to be a contribution for the use of thermography as a methodology for evaluation of skin temperature in the context of orofacial biomechanics. The study aims to identify the oscillations of skin temperature in hemiface’s region of the masseter muscle and estimate the time required to restore the initial temperature after the application of an ice stimulus. Using an infrared camera, a data acquisition protocol was followed with a group of volunteers in a controlled environment. The thermal stimulus involves the use of an ice volume and the skin surface temperature was recorded in two distinct situations, namely without further stimulus and with the addition of a complementary stimulus obtained by a chewing gum. The results shows that recovery is faster with the addition of the stimulus and may guide clinicians regarding the pre and post-operative times with ice therapy, in the presence or absence of mechanical stimulus that increase muscle functions.

This study provides the first description of the larval development of the commercially exploited barnacle Megabalanus azoricus. It describes the changes in larval size and shape as well as the general morphology and duration of each larval stage. Embryos were obtained from gravid specimens collected at Sao Miguel Island and reared through six naupliar stages to the cypris stage in laboratory conditions. The planktotrophic nauplii reached the cypris stage after 14 days of hatching in individual cultures at 20 °C under natural illumination and fed with phytoplankton (Chaetoceros gracilis, Isochrysis sp., and Tetraselmis sp.). The nauplius of M. azoricus has a normal size compared with nauplii of other congeneric species, ranging between the 261 μm (nauplius I) and 912 μm (nauplius VI). This work provides the first description of larvae of the genus Megabalanus for the Portuguese oceanic islands and provides comparisons with congeneric species in other parts of the world.

The giant barnacle Megabalanus azoricus is a popular seafood in the Azores. It is mainly caught in coastal environments and sold for domestic human consumption. This species is a filter feeder and can be used as a biomonitor of trace metal bioavailabilities. To investigate consumption safety, the concentrations of 10 trace metals - As, Cd, Cr, Cu, Mn, Pb, Rb, Se, Sr and Zn - were evaluated in 3 body tissues of M. azoricus from 3 sites on 2 islands. There were no significant differences between the metal loads of the barnacles from the different sites. However, the concentrations of the total trace metal loads revealed significant differences among the tissues (cirrus, muscles and ovaries). The concentrations of some metals in the body were not within the safety levels for consumers, based on the allowable standard levels for crustaceans issued by the European Union and of legislations in several countries. Alarming levels of As and Cd were found. Considering the absence of heavy industry in the region, a non-anthropogenic volcanic source was assumed to be the reason for the observed metal levels. Barnacles, in particular M. azoricus, seem to be useful as bioindicators in this peculiar environment.

In many coastal regions, vegetated habitats (e.g. kelps forests, seagrass beds) play a key role in the structure and functioning of shallow subtidal reef ecosystems, by modifying local environmental conditions and by providing food and habitat for a wide range of organisms. In some regions of the world, however, such idiosyncratic ecosystems are largely absent and are often replaced by less notable ecosystem formers. In the present study, we empirically compared the structure and functioning of two distinct shallow-water habitats present in the Azores: one dominated by smaller frondose brown macroalgae (Dictyotaceae and Halopteris) and one dominated by low-lying turfs. Two replicated areas of each habitat were sampled at two different times of the year, to assess spatial and temporal consistency of results. Habitats dominated by small fronds were significantly (ca. 3 times) more productive (when standardized per algal mass) compared to the turf-dominated habitats, and supported a distinct assemblage (both in terms of composition and abundance) of associated macrofauna. Unlike other well-known and studied vegetated habitats (i.e. kelp forests), however, no effects of habitat were found on the structure of benthonic fish assemblages. Results were spatially and temporally consistent suggesting that, in warmer temperate oceans, habitats dominated by species of smaller frondose brown algae can also play an important role in the structure and functioning of subtidal communities and may, to a certain extent, be considered analogous to other well-known vegetated habitats around the world (i.e. kelp forests, seagrass beds). European Commission BEST Preparatory Action - projecto “ECOSUBVEG em Changes in submersed vegetation: assessing loss in ecosystems services from frondose to depauperate systems dominated by opportunistic vegetation”, ref. 07.032700/2012/635752/SUB/B2. FCT - Fundação para a Ciência e Tecnologia, projecto UID/BIA/00329/2013. CIRN/UAc (Centre of Natural Resources of University of the Azores); CVARG (Centro de Vulcanologia e Avaliação de Riscos Geologicos). info:eu-repo/semantics/publishedVersion

Summary The reproductive biology of Megabalanus azoricus (Pilsbry), the commercially exploited barnacle in the Azores, was studied in an attempt to provide the scientific basis for the sustainable management of this heavily exploited regional marine resource. Both the Ospar Commission and WWF have expressed concern for this species, considering it at risk and in urgent need for scientific study. Barnacles were collected every month from shallow water (≈3 m depth) around Sao Miguel Island from October 2004 to September 2005. Individuals were measured and gonads processed for histology and analysed with a stereological method. M. azoricus has a hermaphroditic reproductive system with separate gonads and it was possible to describe the various stages of gametic maturation in both. Data on fertility were also obtained by egg counts and calculation of the Gonadosomatic index (GSI). Throughout the year, some specimens of M. azoricus can be found that rare mature, but two reproductive peaks were observed, one in...

Re-examination of Megabalanus azoricus from the Azores revealed previously unknown morphological features. Examination of more exemplars also confirmed the description made by Pilsbry in 1916. Morphological features are herein described (e.g. true tergal spur) and morphometric relations are established for the first time, providing a significant contribution towards the management of this important Azorean seafood resource.

The first stage zoea of Microcassiope minor (Dana, 1852) is described, illustrated and compared with thirteen other known Xanthinae. This comparison suggests that M. minor is closely related to another Atlantic Ocean xanthid, Nanocassiope melanodactyla (A. Milne-Edwards, 1867). Such an affinity may indicate that the genus Nanocassiope Guinot, 1967) is not a homogenous taxon.

Global homogenization of biota is underway through worldwide introduction and establishment of non-indigenous (exotic) species. Organisms fouling ship hulls are continually in transit and can affect communities through biodiversity loss and serious damage to economy and public health. In the Azores, for the first time, underwater alien species prospection was conducted in marinas and recreational harbours, at Sao Miguel Island. Populations of three locally previously unknown barnacle species were found: Amphibalanus amphitrite, Amphibalanus eburneus and Perforatus perforatus. These species account for the more than 50% of alien barnacles worldwide that belong to Balanidae family. Hence, some considerations about morphology and life cycle of this family are advanced, discussed and related to their invasive potential.

In the present work, we aimed to explore the potential of two groups of marine invertebrates—sea cucumbers (Echinodermata) and ascidians (Chordata)—as sources of anti-inflammatory, anti-oxidant, and osteogenic compounds with potential to be used as pharmaceuticals and nutraceuticals for the treatment and prevention of chronic diseases. 24 extracts (ethanol, water, and ethyl acetate) from 4 species of sea cucumbers and 4 species of tunicates were produced and screened in vitro for their anti-inflammatory and anti-oxidant activities and in vivo for osteogenic activity through an assay using zebrafish larvae. Our results showed that ethanolic extracts presented anti-oxidant and anti-inflammatory activity, which revealed to be stronger in the ascidians. The osteogenic activity, which provides evidence of the bioactive potential of these organisms in preventing chronic disorders causing low bone density, was found to be strong in one species of ascidians and 3 of holothurians. This study demonstrates the high potential of extracts from these marine organisms for using as nutraceuticals in the prevention of chronic bone disorders.