In Vitro Anti-Inflammatory and Vasculoprotective Effects of Red Cell Extract from the Black Sea Urchin Arbacia lixula.

Sea urchins have emerged as an important source of bioactive compounds with anti-inflammatory and antioxidant properties relevant to human health. Since inflammation is a crucial pathogenic process in the development and progression of atherosclerosis, we here assessed the potential anti-inflammatory and vasculoprotective effects of coelomic red-cell methanolic extract of the black sea urchin Arbacia lixula in an in vitro model of endothelial cell dysfunction. Human microvascular endothelial cells (HMEC-1) were pretreated with A. lixula red-cell extract (10 and 100 µg/mL) before exposure to the pro-inflammatory cytokine tumor necrosis factor (TNF)-α. The extract was non-toxic after 24 h cell treatment and was characterized by antioxidant power and phenol content. The TNF-α-stimulated expression of adhesion molecules (VCAM-1, ICAM-1) and cytokines/chemokines (MCP-1, CCL-5, IL-6, IL-8, M-CSF) was significantly attenuated by A. lixula red-cell extract. This was functionally accompanied by a reduction in monocyte adhesion and chemotaxis towards activated endothelial cells. At the molecular level, the tested extract significantly counteracted the TNF-α-stimulated activation of the pro-inflammatory transcription factor NF-κB. These results provide evidence of potential anti-atherosclerotic properties of A. lixula red-cell extract, and open avenues in the discovery and development of dietary supplements and/or drugs for the prevention or treatment of cardiovascular diseases.

Polymer Chemical Identity as a Key Factor in Microplastic-Insecticide Antagonistic Effects during Embryogenesis of Sea Urchin Arbacia lixula.

As a proxy for pollutants that may be simultaneously present in urban wastewater streams, the effects of two microplastics-polystyrene (PS; 10, 80 and 230 µm diameter) and polymethylmethacrylate (PMMA; 10 and 50 µm diameter)-on fertilisation and embryogenesis in the sea urchin Arbacia lixula with co-exposure to the pyrethroid insecticide cypermethrin were investigated. Synergistic or additive effects were not seen for plastic microparticles (50 mg L-1) in combination with cypermethrin (10 and 1000 µg L-1) based on evaluation of skeletal abnormalities or arrested development and death of significant numbers of larvae during the embryotoxicity assay. This behaviour was also apparent for male gametes pretreated with PS and PMMA microplastics and cypermethrin, where a reduction in sperm fertilisation ability was not evidenced. However, a modest reduction in the quality of the offspring was noted, suggesting that there may be some transmissible damage to the zygotes. PMMA microparticles were more readily taken up than PS microparticles, which could suggest surface chemical identity as potentially modulating the affinity of larvae for specific plastics. In contrast, significantly reduced toxicity was noted for the combination of PMMA microparticles and cypermethrin (100 µg L-1), and may be related to less ready desorption of the pyrethroid than PS, as well as cypermethrin activating mechanisms that result in reduced feeding and hence decreased ingestion of microparticles.

Particle Size Modulates Silver Nanoparticle Toxicity during Embryogenesis of Urchins Arbacia lixula and Paracentrotus lividus.

Silver nanoparticles represent a threat to biota and have been shown to cause harm through a number of mechanisms, using a wide range of bioassay endpoints. While nanoparticle concentration has been primarily considered, comparison of studies that have used differently sized nanoparticles indicate that nanoparticle diameter may be an important factor that impacts negative outcomes. In considering this, the aim of the present study was to determine if different sizes of silver nanoparticles (AgNPs; 10, 20, 40, 60 and 100 nm) give rise to similar effects during embryogenesis of Mediterranean sea urchins Arbacia lixula and Paracentrotus lividus, or if nanoparticle size is a parameter that can modulate embryotoxicity and spermiotoxicity in these species. Fertilised embryos were exposed to a range of AgNP concentrations (1−1000 µg L−1) and after 48 h larvae were scored. Embryos exposed to 1 and 10 µg L−1 AgNPs (for all tested sizes) showed no negative effect in both sea urchins. The smaller AgNPs (size 10 and 20 nm) caused a decrease in the percentage of normally developed A. lixula larvae at concentrations ≥50 µg L−1 (EC50: 49 and 75 µg L−1, respectively) and at ≥100 µg L−1 (EC50: 67 and 91 µg L−1, respectively) for P. lividus. AgNPs of 40 nm diameter was less harmful in both species ((EC50: 322 and 486 µg L−1, for P. lividus and A. lixula, respectively)). The largest AgNPs (60 and 100 nm) showed a dose-dependent response, with little effect at lower concentrations, while more than 50% of larvae were developmentally delayed at the highest tested concentrations of 500 and 1000 µg L−1 (EC50(100 nm); 662 and 529 µg L−1, for P. lividus and A. lixula, respectively. While AgNPs showed no effect on the fertilisation success of treated sperm, an increase in offspring developmental defects and arrested development was observed in A. lixula larvae for 10 nm AgNPs at concentrations ≥50 µg L−1, and for 20 and 40 nm AgNPs at concentrations >100 µg L−1. Overall, toxicity was mostly ascribed to more rapid oxidative dissolution of smaller nanoparticles, although, in cases, Ag+ ion concentrations alone could not explain high toxicity, indicating a nanoparticle-size effect.

Taxonomic reassessment of Tetrapygus niger (Arbacioida, Echinoidea): molecular and morphological evidence support its placement in Arbacia.

The echinoid genus Tetrapygus was initially described by L. Agassiz (1841) based on a single species, Tetrapygus niger Molina, 1782. Since the extensive work conducted by Mortensen (1935), Tetrapygus has received limited taxonomic attention over the past century. Recent discoveries of new fossil species of Arbacia Gray, 1835 from the upper Pliocene of northern Chile revealed striking morphological similarities between the two distinct Arbaciidae genera Arbacia and Tetrapygus. These findings compelled new investigations to evaluate the taxonomic status of these genera. Based on molecular mitochondrial (COI), nuclear (28S), and morphological evidence, Tetrapygus niger is here recovered as the sister species to Arbacia dufresnii, both species forming a clade within the phylogeny of South American species of Arbacia. Consequently, the diagnosis and description of Tetrapygus niger are here revised, and the species is reattributed to Arbacia, as previously proposed by A. Agassiz in Agassiz & Desor (1846) under the species name Arbacia nigra. An emended diagnosis of Arbacia is also proposed in light of these new findings.

Arbacia ballenensis sp. nov. (Echinoidea, Arbacioida): A new species reveals diversification of the genus in Central America.

A new species of Arbacia (Echinoidea, Arbacioida), A. ballenensis sp. nov., from the Late PlioceneEarly Pleistocene of Baja California Norte, Mxico, is described from a series of 62 specimens ranging in size from 6 to 20 mm test diameter. It differs from all extant species in the genus by its high number of interambulacral plates, numerous small interambulacral tubercles, and the size of the interambulacral tags at the peristomial edge. This new species yields new evidence concerning the diversification of the genus along the Pacific coast of North America where previously, only one extinct species was described, Arbacia abiquaensis from the Oligocene of Oregon. A. ballenensis sp. nov. is closely related to the recently described Arbacia quyllur from the late Miocene of Chile. Study of the new species in the context of events such as the mid-Pleistocene Transition helps shed light on patterns of extinction and speciation within the genus.

Cytogenetic and developmental toxicity of bisphenol A and bisphenol S in Arbacia lixula sea urchin embryos.

Bisphenol S (BP-S) is one of the most important substitutes of bisphenol A (BP-A), and its environmental occurrence is predicted to intensify in the future. Both BP-A and BP-S were tested for adverse effects on early life stages of Arbacia lixula sea urchins at 0.1 up to 100 µM test concentrations, by evaluating cytogenetic and developmental toxicity endpoints. Embryonic malformations and/or mortality were scored to determine embryotoxicity (72 h post-fertilization). It has been reported in academic dataset that bisphenols concentration reached µg/L in aquatic environment of heavily polluted areas. We have chosen concentrations ranging from 0.1-100 µM in order to highlight, in particular, BP-S effects. Attention should be paid to this range of concentrations in the context of the evaluation of the toxicity and the ecological risk of BP-S as emerging pollutant. Cytogenetic toxicity was measured, using mitotic activity and chromosome aberrations score in embryos (6 h post-fertilization). Both BP-A and BP-S exposures induced embryotoxic effects from 2.5 to 100 µM test concentrations as compared to controls. Malformed embryo percentages following BP-A exposure were significantly higher than in BP-S-exposed embryos from 0.25 to 100 µM (with a ~5-fold difference). BP-A, not BP-S exhibited cytogenetic toxicity at 25 and 100 µM. Our results indicate an embryotoxic potential of bisphenols during critical periods of development with a potent rank order to BP-A vs. BP-S. Thus, we show that BP-A alternative induce similar toxic effects to BP-A with lower severity.

Acclimation to low pH does not affect the thermal tolerance of Arbacia lixula progeny.

As the ocean warms, the thermal tolerance of marine invertebrates is key to determining their distributional change, where acclimation to low pH may impact the thermal range of optimal development. We compared thermal tolerance of progeny from a low pH-acclimated sea urchin (Arbacia lixula) population from the CO2 vents of Ischia (Italy) and a nearby population living at ambient pH. The percentages of normally developing gastrulae and two-armed larvae were determined across 10 temperatures representing present and future temperature conditions (16-34°C). Vent-acclimated sea urchins showed a greater percentage of normal development at 24 h, with a larger optimal developmental temperature range than control sea urchins (12.3°C versus 5.4°C range, respectively). At 48 h, upper lethal temperatures for 50% survival with respect to ambient temperatures were similar between control (+6.8°C) and vent (+6.2°C) populations. Thus, acclimation to low pH did not impact the broad thermal tolerance of A. lixula progeny. With A. lixula's barrens-forming abilities, its wide thermotolerance and its capacity to acclimate to low pH, this species will continue to be an important ecological engineer in Mediterranean macroalgal ecosystems in a changing ocean.

Unusual polymethylene-interrupted, Δ5 monounsaturated and omega-3 fatty acids in sea urchin (Arbacia punctulata) from the Gulf of Mexico identified by solvent mediated covalent adduct chemical ionization mass spectrometry.

Sea urchin (class Echinoidea) gonads are a prized delicacy in Japan and many other world cultures. The complexity of its fatty acid (FA) profile, particularly minor FA, presents a formidable analytical challenge. We applied solvent mediated (SM) covalent adduct chemical ionization (CACI) tandem mass spectrometry to comprehensive de novo structural and quantitative characterization of the FA profile of Gulf of Mexico Atlantic sea urchin (Arbacia punctulata). >100 FA were detected including many with unusual double bond structure. Gulf sea urchin gonad lipids are rich in Δ5 monounsaturated FA 20:1(5Z) at 2.7% and the polymethylene-interrupted (PMI) diene 20:2(5Z,11Z) at 4.9%, as well as common omega-3 eicosapentaenoic acid (EPA; 5Z, 8Z, 11Z, 14Z, 17Z) at 9.8%±3.1% and arachidonic acid (AA; 5Z, 8Z, 11Z, 14Z) at 6.1%±2.1%. We propose plausible desaturation/elongation-based biochemical pathways for the endogenous production of unusual unsaturates. Unusual unsaturates may modify mammalian signaling and present novel bioactivities.

Simultaneous recording of multiple cellular signaling events by frequency- and spectrally-tuned multiplexing of fluorescent probes.

Fluorescent probes that change their spectral properties upon binding to small biomolecules, ions, or changes in the membrane potential (Vm) are invaluable tools to study cellular signaling pathways. Here, we introduce a novel technique for simultaneous recording of multiple probes at millisecond time resolution: frequency- and spectrally-tuned multiplexing (FASTM). Different from present multiplexing approaches, FASTM uses phase-sensitive signal detection, which renders various combinations of common probes for Vm and ions accessible for multiplexing. Using kinetic stopped-flow fluorimetry, we show that FASTM allows simultaneous recording of rapid changes in Ca2+, pH, Na+, and Vm with high sensitivity and minimal crosstalk. FASTM is also suited for multiplexing using single-cell microscopy and genetically encoded FRET biosensors. Moreover, FASTM is compatible with optochemical tools to study signaling using light. Finally, we show that the exceptional time resolution of FASTM also allows resolving rapid chemical reactions. Altogether, FASTM opens new opportunities for interrogating cellular signaling.

Fertilization and development of Arbacia lixula eggs are affected by osmolality conditions.

The sea urchin Arbacia lixula coexist with Paracentrotus lividus in the Mediterranean, but the two sea urchin species are quite different from each other. Concerning the female gamete, A. lixula eggs are much darker than those of P. lividus due to the characteristic pigmentation. Upon insemination, the fertilization envelope formed by A. lixula eggs is remarkably thinner than that of P. livius eggs, which implies that the cortical organization of the eggs in the two species may be quite different. In this communication, we examined the phenotypic plasticity of A. lixula eggs in the changing osmolality. The plasma membrane, cortical actin cytoskeleton and vesicles are extensively altered in the eggs exposed to 40% seawater for 15 min. When fertilized, the Ca2+ response in these eggs was significantly compromised and the sperm often failed to enter the eggs. Remarkably, the pattern of the Ca2+ response was restored when these eggs were transferring back to the natural seawater before fertilization, while the actin cytoskeleton partially reverted to the original state. Nonetheless, these eggs restored in seawater failed to regain the innate sperm receptivity that allows only one sperm to enter in natural seawater. Thus, the ability to guide monospermic fertilization is lost by water entry into the eggs, and the eggs incorporated either multiple or no sperm. On the other hand, eggs briefly exposed to hypertonic seawater exhibited no evident morphological anomaly. Nonetheless, the monospermic eggs that experienced a brief exposure (15 min) to hypertonic seawater prior to fertilization in natural seawater displayed a subtly altered sperm-induced Ca2+ response and morpho-functional anomaly around the pluteus stage. Our results suggest that A. lixula eggs attain only a limited extent of cytological plasticity, and that the osmolality shock affects the physical nature of the egg surface which in turn affects the developmental programming.

Photoperiod in aquaculture of the sea urchin Arbacia dufresnii (Echinodermata: Echinoidea): Effects on gamete production and maturity / Fotoperiodo en la acuicultura del erizo de mar Arbacia dufresnii (Echinodermata: Echinoidea): El efecto en la producción y madurez de gametas

Introduction: Photoperiod is, together with temperature and food availability, one of the main stimuli in the regulation of gametogenesis in a wide variety of species. Objective: To evaluate the effect of photoperiod on the production of mature gametes in cultured Arbacia dufresnii. Methods: An experiment was carried out with three varying light-dark regimes/treatments: constant light (24 h light), neutral photoperiod (12 h light, 12 h darkness), and constant darkness (24 h darkness). Twenty females were used in each treatment. All were induced to spawn and, ten randomly selected females from each treatment were induced to spawn again after 30 days. After 60 days, spawning was induced in the remaining females. The gametes were collected in filtered seawater, fixed in Davidson solution, quantified and measured per individual in triplicate in a Sedgewick-Rafter chamber. To determine maturation, fertilization success was evaluated 30 minutes after fertilization. Results: Our results showed that in the aquaculture system, after only two months, mature gametes were obtained, and in the neutral light regime there were 10 times more gametes than the number produced in wild sea urchins during the spawning period in question. We also found that with a greater exposure to light, a lower number of mature gametes was produced. Conclusions: This study suggests the viability of the production of mature gametes in a short period of time as regards Arbacia dufresnii.

Introducción: El fotoperiodo es, junto con la temperatura y la disponibilidad de alimentos, uno de los principales estímulos para el desarrollo de la gametogénesis en una amplia variedad de especies. Objetivo: Evaluar el efecto del fotoperiodo en la producción de gametas maduras de Arbacia dufresnii en un sistema de recirculación cerrado para determinar el mejor fotoperiodo para una acuicultura novedosa, enfocada en la producción de gametas con alta concentración de pigmentos para usos biotecnológicos. Métodos: Se realizó un experimento con tres regímenes/tratamientos diferentes de luz y oscuridad: luz constante (luz durante 24 h), fotoperiodo neutro (12 h de luz, 12 h de oscuridad) y oscuridad constante (oscuridad durante 24 h). Se utilizaron veinte hembras en cada tratamiento. Se indujo a todas las hembras a desovar al comienzo del experimento. Después de 30 días, diez hembras seleccionadas al azar de cada tratamiento fueron inducidas a desovar nuevamente. Al final del experimento, después de 60 días, se indujo el desove a las hembras restantes en cada tratamiento. Las gametas se recolectaron en agua de mar filtrada, se fijaron en solución de Davidson, se cuantificaron y midieron por triplicado en una cámara Sedgewick-Rafter. Para determinar la maduración, se evaluó el éxito de la fecundación después de 30 minutos de fertilización, calculando el porcentaje de huevos fertilizados. Resultados: Nuestros resultados muestran que, en el sistema acuícola, en solo dos meses se obtuvieron gametas maduras y casi 10 veces más la cantidad producida por los erizos de mar en su ambiente natural usando el fotoperiodo neutro (12 h luz:12 h oscuridad). También encontramos que la mayor exposición a la luz produce la menor cantidad de gametas maduras. Conclusión: Este estudio sugiere la viabilidad de la producción de gametos maduros en un corto período de tiempo en Arbacia dufresnii.

Comparative study of coelomocytes from Arbacia lixula and Lythechinus variegatus: Cell characterization and in vivo evidence of the physiological function of vibratile cells.

The knowledge on echinoderm coelomocytes has increased in recent years, but researchers still face a complex problem: how to obtain purified cells. Even flow cytometry being useful to address coelomocytes in suspension, the need for a method able to provide isolated cells is still noteworthy. Here, we use Imaging Flow Cytometry (IFC) to characterize the coelomocytes of two sea urchin species - Arbacia lixula and Lytechinus variegatus - and obtain gates to isolate cell populations. Then, we used these gates to study the physiological response of A. lixula coelomocytes during an induced immune challenge with Escherichia coli. An analysis of area and aspect ratio parameters of the flow cytometer allowed the identification of two main cell populations in the coelomic fluid: circular and elongated cells. A combination of this method with nucleus labeling using propidium iodide allowed the determination of gates containing isolated subpopulations of vibratile cells, red spherulocytes, and two phagocytes subpopulations in both species. We observed that during an induced bacterial immune challenge, A. lixula was able to modulate coelomocyte frequencies, increasing the phagocytes and decreasing red spherulocytes and vibratile cells. These results indicate that vibratile cells and red spherulocytes act by immobilizing and stoping bacterial growth, respectively, cooperating with phagocytes in the immune response. The use of IFC was fundamental not only to identify specific gates for the main coelomic subpopulations but also allowed the investigation on how echinoids modulate their physiological responses during immune challenges. Furthermore, we provide the first experimental evidence about the role of vibratile cells, corroborating its involvement with the immune system.

Short-Term Exposure to Storm-Like Scenario Microplastic and Salinity Conditions Does not Impact Adult Sea Urchin (Arbacia punctulata) Physiology.

The effects of microplastic pollution on sea urchins has received little attention despite their ecological and economical importance. This is the first study to focus on adult sea urchins (Arbacia punctulata). These organisms were exposed to storm-like sediment resuspension of microplastic concentrations (9-µm polystyrene 25,000 spheres L-1) combined with salinity reductions (salinity 25 vs. 33) associated with high precipitation. Urchins were exposed to these parameters for 24 h before assessing righting times and for 48 h before assessing oxygen consumption rates. No significant impacts on urchin physiology were observed showing resilience to short-term exposures of storm-like induced microplastics and salinity. No microplastic particles blocked the madreporite pores indicating the active removal of particles by cilia and pedicellariae. Gut tissue samples indicated consumption of microplastics. Studies on more species are urgently required to determine their responses to plastic pollution to inform management decision-making processes.

Underwater high frequency noise: Biological responses in sea urchin Arbacia lixula (Linnaeus, 1758).

Marine life is extremely sensitive to the effects of environmental noise due to its reliance on underwater sounds for basic life functions, such as searching for food and mating. However, the effects on invertebrate species are not yet fully understood. The aim of this study was to determine the biochemical responses of Arbacia lixula exposed to high-frequency noise. Protein concentration, enzyme activity (esterase, phosphatase and peroxidase) and cytotoxicity in coelomic fluid were compared in individuals exposed for three hours to consecutive linear sweeps of 100 to 200 kHz lasting 1 s, and control specimens. Sound pressure levels ranged between 145 and 160 dB re 1µPa. Coelomic fluid was extracted and the gene and protein expression of HSP70 with RT-PCR was evaluated on coelomocytes. A significant change was found in enzyme activity and in the expression of the HSP70 gene and protein compared to the control. These results suggested that high-frequency stimuli elicit a noise-induced physiological stress response in A. lixula, confirming the vulnerability of this species to acoustic exposure. Furthermore, these findings provide the first evidence that cell-free coelomic fluid can be used as a signal to evaluate noise exposure in marine invertebrates.

Absolute proteomic quantification reveals design principles of sperm flagellar chemosensation.

Cilia serve as cellular antennae that translate sensory information into physiological responses. In the sperm flagellum, a single chemoattractant molecule can trigger a Ca2+ rise that controls motility. The mechanisms underlying such ultra-sensitivity are ill-defined. Here, we determine by mass spectrometry the copy number of nineteen chemosensory signaling proteins in sperm flagella from the sea urchin Arbacia punctulata. Proteins are up to 1,000-fold more abundant than the free cellular messengers cAMP, cGMP, H+ , and Ca2+ . Opto-chemical techniques show that high protein concentrations kinetically compartmentalize the flagellum: Within milliseconds, cGMP is relayed from the receptor guanylate cyclase to a cGMP-gated channel that serves as a perfect chemo-electrical transducer. cGMP is rapidly hydrolyzed, possibly via "substrate channeling" from the channel to the phosphodiesterase PDE5. The channel/PDE5 tandem encodes cGMP turnover rates rather than concentrations. The rate-detection mechanism allows continuous stimulus sampling over a wide dynamic range. The textbook notion of signal amplification-few enzyme molecules process many messenger molecules-does not hold for sperm flagella. Instead, high protein concentrations ascertain messenger detection. Similar mechanisms may occur in other small compartments like primary cilia or dendritic spines.

Effects of elevated temperature on gonadal functions, cellular apoptosis, and oxidative stress in Atlantic sea urchin Arbacia punculata.

Increasing seawater temperature affects growth, reproduction and development in marine organisms. In this study, we examined the effects of elevated temperatures on reproductive functions, heat shock protein 70 (HSP70) and nitrotyrosine protein (NTP, an indicator of reactive nitrogen species) expressions, protein carbonyl (PC, an indicator of oxidative stress) contents, cellular apoptosis, and coelomic fluid (CF) conditions in Atlantic sea urchin. Sea urchins were housed in six aquaria with control (24 °C) and elevated temperatures (28 °C and 32 °C) for a 7-day period. After exposure, sea urchins exhibited decreased percentages of gametes (eggs/sperm), as well as increased HSP70 and NTP expressions in eggs and spermatogenic cells, increased gonadal apoptosis, and decreased CF pH compared to controls. PC contents were also significantly increased in gonadal tissues at higher temperatures. These results suggest that elevated temperature acidifies CF, increases oxidative stress and gonadal apoptosis, and results in impairment of reproductive functions in sea urchins.

No-take marine reserves control the recovery of sea urchin populations after mass mortality events.

Understanding how no-take zones (NTZs) shape the population dynamics of key herbivores is crucial for the conservation and management of temperate benthic communities. Here, we examine the recovery patterns of sea urchin populations following a high-intensity storm under contrasting protection regimes in the NW Mediterranean Sea. We found significant differences in the recovery trends of Paracentrotus lividus abundance and biomass in the five years following the storm. The P. lividus populations outside the NTZ recovered faster than the populations inside the NTZ, revealing that predation was the main factor controlling the sea urchin populations inside the NTZ during the study period. Arbacia lixula reached the highest abundance and biomass values ever observed outside the NTZ in 2016. Our findings reveal that predation can control the establishment of new sea urchin populations and emphasize top-down control in NTZs, confirming the important role of fully protected areas in the structure of benthic communities.

Spatio-temporal patterns of genetic variation in Arbacia lixula, a thermophilous sea urchin in expansion in the Mediterranean.

The genetic structure of 13 populations of the amphiatlantic sea urchin Arbacia lixula, as well as temporal genetic changes in three of these localities, were assessed using ten hypervariable microsatellite loci. This thermophilous sea urchin is an important engineer species triggering the formation of barren grounds through its grazing activity. Its abundance seems to be increasing in most parts of the Mediterranean, probably favoured by warming conditions. Significant genetic differentiation was found both spatially and temporally. The main break corresponded to the separation of western Atlantic populations from those in eastern Atlantic and the Mediterranean Sea. A less marked, but significant differentiation was also found between Macaronesia (eastern Atlantic) and the Mediterranean. In the latter area, a signal of differentiation between the transitional area (Alboran Sea) and the rest of the Mediterranean was detected. However, no genetic structure is found within the Mediterranean (excluding Alboran) across the Siculo-Tunisian Strait, resulting from either enough gene flow to homogenize distance areas or/and a recent evolutionary history marked by demographic expansion in this basin. Genetic temporal variation at the Alboran Sea is as important as spatial variation, suggesting that temporal changes in hydrological features can affect the genetic composition of the populations. A picture of genetic homogeneity in the Mediterranean emerges, implying that the potential expansion of this keystone species will not be limited by intraspecific genetic features and/or potential impact of postulated barriers to gene flow in the region.

Comparative toxicity evaluation of targeted anticancer therapeutics in embryonic zebrafish and sea urchin models.

Cancer drug resistance and poor selectivity towards cancer cells demand the constant search for new therapeutics. PI3K-Akt-mTOR and RAS-MAPK-ERK signaling pathways are key mechanisms involved in cell survival, proliferation, differentiation, and metabolism and their deregulation in cancer can promote development of therapy resistance. We investigated the effects of targeted inhibitors (wortmannin, GSK690693, AZD2014 and tipifarnib) towards these two pathways on early zebrafish and sea urchin development to assess their toxicity in normal, fast proliferating cells. PI3K inhibitor wortmannin and RAS inhibitor tipifarnib displayed highest toxicity while GSK690693, a pan-Akt kinase inhibitor, exhibited a less significant impact on embryo survival and development. Moreover, inhibition of the upstream part of the PI3K-Akt-mTOR pathway (wortmannin/GSK690693 co-treatment) produced a synergistic effect and impacted zebrafish embryo survival and development at much lower concentrations. Dual mTORC1/mTORC2 inhibitor AZD2014 showed no considerable effects on embryonic cells of zebrafish in concentrations substantially toxic in cancer cells. AZD2014 also caused the least prominent effects on sea urchin embryo development compared to other inhibitors. Significant toxicity of AZD2014 in human cancer cells, its capacity to sensitize resistant cancers, lower antiproliferative activity against human normal cell lines and fast proliferating embryonic cells could make this agent a promising candidate for anticancer therapy.

Copper oxide nanoparticles induce the transcriptional modulation of oxidative stress-related genes in Arbacia lixula embryos.

Copper oxide nanoparticles (CuO NPs) are widely used in various industrial applications, i.e. semiconductor devices, batteries, solar energy converter, gas sensor, microelectronics, heat transfer fluids, and have been recently recognized as emerging pollutants of increasing concern for human and marine environmental health. Therefore, the toxicity of CuO NPs needs to be thoroughly understood. In this study, we evaluated the potential role of oxidative stress in CuO NP toxicity by exploring the molecular response of Arbacia lixula embryos to three CuO NP concentrations (0.7, 10, 20 ppb) by investigating the transcriptional patterns of oxidative stress-related genes (catalase and superoxide dismutase) and metallothionein, here cloned and characterized for the first time. Time- and concentration-dependent changes in gene expression were detected in A. lixula embryos exposed to CuO NPs, up to pluteus stage (72 h post-fertilization, hpf), indicating that oxidative stress is one of the toxicity mechanisms for CuO NPs. These findings provide new insights into the comprehension of the molecular mechanisms underlying copper nanoparticle toxicity in A. lixula sea urchin and give new tools for monitoring of aquatic areas, thus corroborating the suitability of this embryotoxicity assay for future evaluation of impacted sites.