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Anemonia sulcata may be a source of marine natural products (MNPs) due to the antioxidant and antitumor activity of its crude homogenates shown in vitro in colon cancer cells. A bioguided chromatographic fractionation assay of crude Anemonia sulcata homogenates with and without its symbiont Symbiodinium was performed to characterize their bioactive composition and further determine their biological potential for the management of colorectal cancer (CRC). The 20% fractions retained the in vitro antioxidant activity previously reported for homogenates. As such, activation of antioxidant and detoxifying enzymes was also evaluated. The 40% fractions showed the greatest antiproliferative activity in T84 cells, synergistic effects with 5-fluoruracil and oxaliplatin, overexpression of apoptosis-related proteins, cytotoxicity on tumorspheres, and antiangiogenic activity. The predominantly polar lipids and toxins tentatively identified in the 20% and 40% fractions could be related to their biological activity in colon cancer cells although further characterizations of the active fractions are necessary to isolate and purify the bioactive compounds. [ABSTRACT FROM AUTHOR]

For the first time, we studied the morpho-functional characteristics of amoebocytes of the sea anemone Anemonia viridis. A histochemical approach was adopted and two subpopulations of amoebocytes from mesoglea were observed: granulocytes and hyalinocytes. Granulocytes showed a high number of cytoplasmatic granules, while hyalinocytes had no or few granules. Amoebocytes showed both round and spreading shapes and were divided in basophil, neutrophil or acidophil cells. Amoebocytes actively phagocytized yeast cells and produced superoxide anion. The presence of hydrolytic enzymes in amoebocytes was also investigated. Positive cells to acid phosphatase, acid esterase and non-specific esterase were found, with granulocytes and hyalinocytes that did not show different responses in term of positivity. Although preliminary, the results of this study can help to understand the features and immune role of A. viridis amoebocytes and the development of defense strategies in multicellular organisms. Moreover, this study lay the foundations for future more in-depth studies, including those at the ultrastructural level. [ABSTRACT FROM AUTHOR]

Anemonia sulcata may be a source of marine natural products (MNPs) due to the antioxidant and antitumor activity of its crude homogenates shown in vitro in colon cancer cells. A bioguided chromatographic fractionation assay of crude Anemonia sulcata homogenates with and without its symbiont Symbiodinium was performed to characterize their bioactive composition and further determine their biological potential for the management of colorectal cancer (CRC). The 20% fractions retained the in vitro antioxidant activity previously reported for homogenates. As such, activation of antioxidant and detoxifying enzymes was also evaluated. The 40% fractions showed the greatest antiproliferative activity in T84 cells, synergistic effects with 5-fluoruracil and oxaliplatin, overexpression of apoptosis-related proteins, cytotoxicity on tumorspheres, and antiangiogenic activity. The predominantly polar lipids and toxins tentatively identified in the 20% and 40% fractions could be related to their biological activity in colon cancer cells although further characterizations of the active fractions are necessary to isolate and purify the bioactive compounds.

The sexual reproduction of the sea anemone, Anemonia sulcata (Pennant, 1777), was studied for the first time at Cape Mazari in the Moroccan Mediterranean coast over an annual cycle from May 2017 to May 2018. A total of 330 individuals of A. sulcata were sampled during 11 months. The sampled individuals were measured, weighed, and fixed for histological analyses. The pedal disc diameter (PDD) of the species ranged from 1.2 cm to 6.6 cm with a mean value of 2.7 ± 0.6 cm. The weight of the species varied from 2.6 g to 94.0 g with a mean value of 19.9 ± 12.7 g. Results of histological analyses indicated a sex ratio of 2.3 female: 1 male. Gonads were observed generally throughout the study period and mature ones were encountered from February to June. Five possible maturity stages of A. sulcata for each gender were recorded. Based on this study, the exploitation of A. sulcata can be managed in the Moroccan Mediterranean coast by the application of the biological rest period from February to June as well as monitoring its weight [ABSTRACT FROM AUTHOR]

Contamination from metals as a result of anthropic action in coastal marine areas is increasing and it is therefore of great importance to have an appropriate follow-up program in these areas. Samples were taken in the intertidal in 6 sites, of which 5 were in the island of Tenerife and one in the island of Lanzarote (Canary Islands, Spain); in all sites, 15 anemone samples were collected. Twenty metals were analyzed by ICP-OES. The results show the highest concentrations of contamination in the area of Los Silos and Puerto del Carmen for the anthropogenic metals Cr, B, Ni, Cu, Pb, Cd, Fe, Li, and V; Los Silos shows Pb 25.264 ± 27.185 mg/kg and Cd 0.058 ± 0.050 mg/kg, which are high compared with the other sites and indicate that the area may be exposed to a high degree of contamination. Compared with other studies, Los Silos has the same or greater concentrations of anthropogenic metals than areas of greater pollution, such as the Mediterranean Sea or the Indian Ocean. For these reasons, Anemonia sulcata can be used as a bioindicator of anthropogenic contamination and it has become a useful organism for environmental pollution monitoring studies. [ABSTRACT FROM AUTHOR]

Recently, invertebrate marine species have been investigated for the presence of natural products with antitumor activity. We analyzed the invertebrate Anemonia sulcata with (W) and without (W/O) the presence of its microalgal symbiont Symbiodinium as a source of bioactive compounds that may be applied in the therapy and/or prevention of colorectal cancer (CRC). Animals were mechanically homogenized and subjected to ethanolic extraction. The proximate composition and fatty acid profile were determined. In addition, an in vitro digestion was performed to study the potentially dialyzable fraction. The antioxidant and antitumor activity of the samples and the digestion products were analyzed in CRC cells in vitro. Our results show a high concentration of polyunsaturated fatty acid in the anemone and a great antioxidant capacity, which demonstrated the ability to prevent cell death and a high antitumor activity of the crude homogenates against CRC cells and multicellular tumor spheroids, especially W/O symbiont. These preliminary results support that Anemonia sulcata could be a source of bioactive compounds with antioxidant and antitumor potential against CRC and that the absence of its symbiont may enhance these properties. Further studies will be necessary to define the bioactive compounds of Anemonia sulcata and their mechanisms of action.

Marine-derived fungi are an important source of many valuable compounds with original structures and diverse physico-chemical properties. In this work, the metabolomic profile of a strain of Penicillium brevicompactum, recovered from a snakelocks sea anemone (Anemonia sulcata), was investigated through the parallel application of LC-ESI-HRMS, GC-MS, and NMR. Our strategy allowed the identification of mycophenolic acid, brevianamide A, and several compounds belonging to the thiosilvatins. Among the latter, five products are reported for the first time in this species. The main product of this series, cis-bis(methylthio)silvatin, was also tested for antiproliferative activity on both cancer and non-tumoral colon cell lines.

Aim: Sodium houttuyfonate (SH), a chemical compound originating from Houttuynia cordata, has been reported to have anti-inflammatory, antibacterial, and antifungal effects, as well as cardioprotective effects. In this study, we investigated the effects of SH on cardiac electrophysiology, because to the best of our knowledge, this issue has not been previously investigated. Methods: We used the whole-cell patch-clamp technique to explore the effects of SH on peak sodium current (INa.P) and late sodium current (INa.L) in isolated rabbit ventricular myocytes. To test the drug safety of SH, we also investigated the effect of SH on rapidly activated delayed rectifier potassium current (IKr). Results: SH (1, 10, 50, and 100 μmol/L) inhibited INa.P in a concentration-dependent manner with an IC50 of 78.89 μmol/L. In addition, SH (100 μmol/L) accelerated the steady state inactivation of INa.P. Moreover, 50 and 100 μmol/L SH inhibited Anemonia sulcata toxin II (ATX II)-increased INa.L by 30.1 and 57.1%, respectively. However, SH (50 and 100 μmol/L) only slightly affected IKr. Conclusions: The inhibitory effects of SH on ATX II-increased INa.L may underlie the electrophysiological mechanisms of the cardioprotective effects of SH; SH has the potential to be an effective and safe antiarrhythmic drug. [ABSTRACT FROM AUTHOR]

Abstract: Traditional aquaculture produces wastewater with high nutrient and organic matter concentrations. Poly‐culture can improve this problem including “extractive species” such as sea cucumbers along with the primary species. The influence of sea cucumbers on transparent exopolymer particles (TEP) (i.e. biofilm precursors) has not been previously explored. Here, we monitored during 1 year the concentration of nutrients, total organic carbon (TOC), particulate organic matter (POM), TEP, chlorophyll‐a and bacteria in two tanks of 50,000 L. One tank only contained Anemonia sulcata, whereas the other tank also included holothurians. To complement these time‐series, we performed three short‐term experiments in smaller (300 L) tanks. Three tanks contained A. sulcata plus Holothuria tubulosa (+H treatment) and other four tanks contained only A. sulcata (−H treatment). In the time‐series, we found that the concentration of ammonium, nitrate, TOC, POM, TEP and bacteria in the effluent of the tank with holothurians was lower than in the effluent of the tank without holothurians. The three experiments confirmed that the holothurians reduced significantly nitrates, bacterial abundance and TEP concentration. Therefore, these invertebrates can control bacterial proliferation and prevent biofilm formation minimizing likely the risk of outbreak of pathogenic bacteria and improving the hygiene of the tanks. [ABSTRACT FROM AUTHOR]

δ 15 N concentration can reveal the anthropogenic pollution of an ecosystem. To study this pollution level, this isotope can be analyzed from water or animal tissue. Sewage pipes pour anthropic waters, rich in δ 15 N, to the sea. In the sewage pipes surroundings live several organisms, like the cnidarian Anemonia sulcata (Pennant, 1777). As anemones have zooxanthellae, they incorporate nitrogen from the environment. To test for the possible effects of wastewater effluents on the coastal ecosystem through the content of the isotope δ 15 N, A. sulcata specimens were collected near a sewage pipe in North Tenerife (Spain), in contiguous zones to look for a δ 15 N gradient. The obtained results show that there is a gradient of δ 15 N concentration, and the highest levels were found nearest the sewage pipe (mean = 5 δ 15 N ‰) and the lowest in the control zone (mean = 3.26 δ 15 N ‰). Being so, A. sulcata has proven to be a sensitive organism whose δ 15 N concentration can be used as bioindicator for pollution and human involvement in the ecosystem in which it lives. [ABSTRACT FROM AUTHOR]

The mitochondrial genomes of sea anemones are dynamic in structure. Invasion by genetic elements, such as self-catalytic group I introns or insertion-like sequences, contribute to sea anemone mitochondrial genome expansion and complexity. By using next generation sequencing we investigated the complete mtDNAs and corresponding transcriptomes of the temperate sea anemone Anemonia viridis and its closer tropical relative Anemonia majano . Two versions of fused homing endonuclease gene ( HEG ) organization were observed among the Actiniidae sea anemones; in-frame gene fusion and pseudo-gene fusion. We provided support for the pseudo-gene fusion organization in Anemonia species, resulting in a repressed HEG from the COI-884 group I intron. orfA , a putative protein-coding gene with insertion-like features, was present in both Anemonia species. Interestingly, orfA and COI expression were significantly up-regulated upon long-term environmental stress corresponding to low seawater pH conditions. This study provides new insights to the dynamics of sea anemone mitochondrial genome structure and function. [ABSTRACT FROM AUTHOR]

Cnidarians harbor a variety of small regulatory RNAs that include microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), but detailed information is limited. Here, we report the identification and expression of novel miRNAs and putative piRNAs, as well as their genomic loci, in the symbiotic sea anemone Anemonia viridis. We generated a draft assembly of the A. viridis genome with putative size of 313 Mb that appeared to be composed of about 36% repeats, including known transposable elements. We detected approximately equal fractions of DNA transposons and retrotransposons. Deep sequencing of small RNA libraries constructed from A. viridis adults sampled at a natural CO2 gradient off Vulcano Island, Italy, identified 70 distinct miRNAs. Eight were homologous to previously reported miRNAs in cnidarians, whereas 62 appeared novel. Nine miRNAs were recognized as differentially expressed along the natural seawater pH gradient. We found a highly abundant and diverse population of piRNAs, with a substantial fraction showing ping-pong signatures. We identified nearly 22% putative piRNAs potentially targeting transposable elements within the A. viridis genome. The A. viridis genome appeared similar in size to that of other hexacorals with a very high divergence of transposable elements resembling that of the sea anemone genus Exaiptasia. The genome encodes and expresses a high number of small regulatory RNAs, which include novel miRNAs and piRNAs. Differentially expressed small RNAs along the seawater pH gradient indicated regulatory gene responses to environmental stressors. [ABSTRACT FROM AUTHOR]

Gene family encoding translationally controlled tumour protein (TCTP) is defined as highly conserved among organisms; however, there is limited knowledge of non-bilateria. In this study, the first TCTP homologue from anthozoan was characterised in the Mediterranean Sea anemone, Anemonia viridis. The release of the genome sequence of Acropora digitifera, Exaiptasia pallida, Nematostella vectensis and Hydra vulgaris enabled a comprehensive study of the molecular evolution of TCTP family among cnidarians. A comparison among TCTP members from Cnidaria and Bilateria showed conserved intron exon organization, evolutionary conserved TCTP signatures and 3D protein structure. The pattern of mRNA expression profile was also defined in A. viridis. These analyses revealed a constitutive mRNA expression especially in tissues with active proliferation. Additionally, the transcriptional profile of A. viridis TCTP (AvTCTP) after challenges with different abiotic/biotic stresses showed induction by extreme temperatures, heavy metals exposure and immune stimulation. These results suggest the involvement of AvTCTP in the sea anemone defensome taking part in environmental stress and immune responses. [ABSTRACT FROM AUTHOR]

We investigated the antibacterial activity of Anemonia sulcata (Cnidaria, Anthozoa) tentacle and body acidic extracts. Biochemical purification consisted of first step on solid phase Sep-Pak C8 column followed by several HPLC runs on C18 column using different conditions. Anti-Micrococcus lysodeikticus activity has been detected in 40 % acetonitrile fractions. The resulting purified molecule from tentacles had a molecular mass determined by MALDI-TOF mass spectrum of 4946,299 Da and has been completely sequenced. Its aa sequence revealed identity with the Neurotoxin 2 (ATX-II), a Na+ channel blocking toxins. Consequently, ATX-II appeared to display a dual role as toxin and as antibacterial.

Gene family encoding allograft inflammatory factor-1 (AIF-1) is well conserved among organisms; however, there is limited knowledge in lower organisms. In this study, the first AIF-1 homologue from cnidarians was identified and characterised in the sea anemone Anemonia viridis . The full-length cDNA of AvAIF-1 was of 913 bp with a 5′ -untranslated region (UTR) of 148 bp, a 3′-UTR of 315 and an open reading frame (ORF) of 450 bp encoding a polypeptide with149 amino acid residues and predicted molecular weight of about 17 kDa. The predicted protein possesses evolutionary conserved EF hand Ca 2+ binding motifs, post-transcriptional modification sites and a 3D structure which can be superimposed with human members of AIF-1 family. The AvAIF-1 transcript was constitutively expressed in all tested tissues of unchallenged sea anemone, suggesting that AvAIF-1 could serve as a general protective factor under normal physiological conditions. Moreover, we profiled the transcriptional activation of AvAIF-1 after challenges with different abiotic/biotic stresses showing induction by warming conditions, heavy metals exposure and immune stimulation. Thus, mechanisms associated to inflammation and immune challenges up-regulated AvAIF-1 mRNA levels. Our results suggest its involvement in the inflammatory processes and immune response of A. viridis. [ABSTRACT FROM AUTHOR]

Marine benthic organisms can be used as indicators of the quality of environmental status and as monitoring tools to detect natural or anthropogenic perturbations. In temperate waters, metabolic and biochemical responses may be governed by physiological changes driven by seasonal factors. Gathering baseline information on the mechanisms underlying seasonal acclimation patterns is therefore a critical step towards the understanding of the physiological responses of biological indicators. In poikilothermic metazoans, the production of regulatory metabolic enzymes can be used as tools for deciphering the acclimation potential. The aim of this study was to characterize the natural seasonal variability in biometric traits and enzymatic biomarkers over a 12-month period in the sea anemone, Anemonia sulcata (Anthozoa, Cnidaria), from two areas with different environmental and anthropic impacts. Seasonality and site factors affected enzymatic kinetics at tentacle levels, while seasonality, site and biometrical characteristics modulated the same enzymes in the body column of A. sulcata. The results showed a decrease in enzymatic activity in summer and an increased synthesis of enzymes in the late fall and winter months. The interaction between biometric features and temperature appeared significant for mesolittoral sea anemones but not for infralittoral specimens. This study showed seasonal patterns of variations of biochemical responses in A. sulcata, suggesting that this species, an abundant member of shallow rocky bottom communities in temperate European seas, may represent a sensitive bio-indicator organism worth considering for further ecological studies on climate change as well as for environmental monitoring programs. [ABSTRACT FROM AUTHOR]

Marine invertebrates have been attracting the attention of researchers for their application in nutrition, agriculture, and the pharmaceutical industry, among others. Concerning sea anemones (Cnidaria), little is known regarding their metabolic profiles and potential value as a source of pharmacologically-active agents. In this work, the chemical profiles of two species of sea anemones Actinia equina and Anemonia sulcata, were studied by high-performance liquid chromatography with diode-array detection (HPLC-DAD) and its impact upon immune and gastric cells was evaluated. In both species, the methylpyridinium alkaloid homarine was the major compound in aqueous extracts. The extracts were effective in reducing lipopolysaccharide (LPS)-induced levels of nitric oxide (NO) and intracellular reactive oxygen species (ROS) in a macrophage model of inflammation. Both the extracts and the alkaloid homarine were effective in inhibiting phospholipase A2 (PLA2), a pivotal enzyme in the initial steps of the inflammatory cascade. In order to mimic the oral consumption of these extracts; their effect upon human gastric cells was evaluated. While no caspase-9 activation was detected, the fact that the endoplasmic reticulum-resident caspase-4, and also caspase-3, were activated points to a non-classical mechanism of apoptosis in human gastric cells. This work provides new insights on the toxicity and biological potential of sea anemones increasingly present in human nutrition. [ABSTRACT FROM AUTHOR]

The present work identifies the decapod crustaceans that associate with Anemonia sulcata on the southwestern Atlantic coast of Portugal and characterises their host use pattern. It determines whether the anemone is monopolised by any species, resulting in the exclusion of conspecifics or other decapods and, under laboratory conditions, it evaluates the degree of association between each species and A. sulcata. From all sampled anemones, 79% harboured at least 1 decapod crustacean, with the majority displaying either one or two specimens (32 and 24%, respectively). The most abundant species were the shrimp Periclimenes sagittifer and the crab Inachus phalangium (representing 36 and 31% of collected specimens, respectively), which displayed lasting associations and were commonly recorded among the tentacles of the host. The species Eualus occultus, E. complex cranchii, Clibanarius erythropus, Maja brachydactyla, Pilumnus hirtellus and Polybius (Necora) puber displayed short-term associations, were mainly present on the substratum near the base, and avoided the tentacles of A. sulcata. Periclimenes sagittifer and I. phalangium were only recorded alone or in heterosexual pairs, appearing to efficiently defend their host against conspecifics. The majority of recorded species only seem to temporarily associate with A. sulcata, in order to seek protection from predators when other shelters are unavailable.

The evolution of multicellular organisms was marked by adaptations to protect against pathogens. The mechanisms for discriminating the ’’self’’ from ’’non-self” have evolved into a long history of cellular and molecular strategies, from damage repair to the co-evolution of host-pathogen interactions. We investigated the inflammatory response in Anemonia sulcata (Cnidaria: Anthozoa) following injection of substances that varied in type and dimension, and observed clear, strong and specific reactions, especially after injection of Escherichia coli and Vibrio alginolyticus . Moreover, we analyzed enzymatic activity of protease, phosphatase and esterase, showing how the injection of different bacterial strains alters the expression of these enzymes and suggesting a correlation between the appearance of the inflammatory reaction and the modification of enzymatic activities. Our study shows for the first time, a specific reaction and enzymatic responses following injection of bacteria in a cnidarian. [ABSTRACT FROM AUTHOR]

Pathological enhancement of late Na+ current (INa) can potentially modify intracellular ion homeostasis and contribute to cardiac dysfunction. We tested the hypothesis that modulation of late INa can be a source of intracellular Na+ ([Na+]i) overload. Late INa was enhanced by exposing rabbit ventricular myocytes to Anemonia sulcata toxin II (ATX-II) and measured using whole cell patch-clamp technique. [Na+]i was determined with fluorescent dye Asante NaTRIUM Green-2 AM. Pacing-induced changes in the dye fluorescence measured at 37°C were more pronounced in ATX-II-treated cells than in control (dye washout prevented calibration). At 22-24°C, resting [Na+]i was 6.6 ± 0.8 mM. Treatment with 5 nM ATX-II increased late INa 8.7-fold. [Na+]i measured after 2 min of electrical stimulation (1 Hz) was 10.8 ± 1.5 mM and 22.1 ± 1.6 mM (P < 0.001) in the absence and presence of 5 nM ATX-II, respectively. Inhibition of late INa with GS-967 (1 µM) prevented Na+i accumulation. A strong positive correlation was observed between the late INa and the pacing-induced increase of [Na+]i (R² = 0.88) and between the rise in [Na+]i and the increases in cytosolic Ca2+ (R² = 0.96). ATX-II, tetrodotoxin, or GS-967 did not affect [Na+]i in quiescent myocytes suggesting that late INa was solely responsible for triggering the ATX-II effect on [Na+]i. Experiments with pinacidil and E4031 indicate that prolongation of the action potential contributes to as much as 50% of the [Na+]i overload associated with the increase in late INa caused by ATX-II. Enhancement of late INa can cause intracellular Na+ overload in ventricular myocytes. [ABSTRACT FROM AUTHOR]

Cnidarian–dinoflagellate symbiosis mainly relies on nutrient recycling, thus providing both partners with a competitive advantage in nutrient-poor waters. Essential processes related to lipid metabolism can be influenced by various factors, including hyperthermal stress. This can affect the lipid content and distribution in both partners, while contributing to symbiosis disruption and bleaching. In order to gain further insight into the role and distribution of lipids in the cnidarian metabolism, we investigated the lipid composition of the sea anemone Anemonia viridis and its photosynthetic dinoflagellate endosymbionts ( Symbiodinium ). We compared the lipid content and fatty acid profiles of the host cellular layers, non-symbiotic epidermal and symbiont-containing gastrodermal cells, and those of Symbiodinium , in a mass spectrometry-based assessment. Lipids were more concentrated in Symbiodinium cells, and the lipid class distribution was dominated by polar lipids in all tissues. The fatty acid distribution between host cell layers and Symbiodinium cells suggested potential lipid transfers between the partners. The lipid composition and distribution was modified during short-term hyperthermal stress, mainly in Symbiodinium cells and gastrodermis. Exposure to elevated temperature rapidly caused a decrease in polar lipid C18 unsaturated fatty acids and a strong and rapid decrease in the abundance of polar lipid fatty acids relative to sterols. These lipid indicators could therefore be used as sensitive biomarkers to assess the physiology of symbiotic cnidarians, especially the effect of thermal stress at the onset of cnidarian bleaching. Overall, the findings of this study provide some insight on key lipids that may regulate maintenance of the symbiotic interaction. [ABSTRACT FROM AUTHOR]

Using a partial transcriptome of the snakelocks anemone ( Anemonia sulcata ) we identify toxin gene candidates that were incorrectly assembled into several Trinity components. Our approach recovers hidden diversity found within some toxin gene families that would otherwise go undetected when using Trinity, a widely used program for venom-focused transcriptome reconstructions. Unidentified hidden transcripts may significantly impact conclusions made regarding venom composition (or other multi-copy conserved genes) when using Trinity or other de novo assembly programs. [ABSTRACT FROM AUTHOR]

We investigated the antibacterial activity of Anemonia sulcata (Cnidaria, Anthozoa) tentacle and body acidic extracts. Biochemical purification consisted of first step on solid phase Sep-Pak C8 column followed by several HPLC runs on C18 column using different conditions. Anti-Micrococcus lysodeikticus activity has been detected in 40 % acetonitrile fractions. The resulting purified molecule from tentacles had a molecular mass determined by MALDI-TOF mass spectrum of 4946,299 Da and has been completely sequenced. Its aa sequence revealed identity with the Neurotoxin 2 (ATX-II), a Na+ channel blocking toxins. Consequently, ATX-II appeared to display a dual role as toxin and as antibacterial.

Gene family encoding small Heat-Shock Proteins (sHSPs containing α-crystallin domain) are found both in prokaryotic and eukaryotic organisms; however, there is limited knowledge of their evolution. In this study, two small HSP genes termed AvHSP28.6 and AvHSP27, both organized in one intron and two exons, were characterised in the Mediterranean snakelocks anemone Anemonia viridis. The release of the genome sequence of Hydra magnipapillata and Nematostella vectensis enabled a comprehensive study of the molecular evolution of α-crystallin gene family among cnidarians. Most of the H. magnipapillata sHSP genes share the same gene organization described for AvHSP28.6 and AvHSP27, differing from the sHSP genes of N. vectensis which mainly show an intronless architecture. The different genomic organization of sHSPs, the phylogenetic analyses based on protein sequences, and the relationships among Cnidarians, suggest that the A.viridis sHSPs represent the common ancestor from which H. magnipapillata genes directly evolved through segmental genome duplication. Additionally retroposition events may be considered responsible for the divergence of sHSP genes of N. vectensis from A. viridis. Analyses of transcriptional expression profile showed that AvHSP28.6 was constitutively expressed among different tissues from both ectodermal and endodermal layers of the adult sea anemones, under normal physiological conditions and also under different stress condition. Specifically, we profiled the transcriptional activation of AvHSP28.6 after challenges with different abiotic/biotic stresses showing induction by extreme temperatures, heavy metals exposure and immune stimulation. Conversely, no AvHSP27 transcript was detected in such dissected tissues, in adult whole body cDNA library or under stress conditions. Hence, the involvement of AvHSP28.6 gene in the sea anemone defensome is strongly suggested. [ABSTRACT FROM AUTHOR]

Highlights: [•] A Drosophila sodium channel, DmNav7-1, exhibits a unique persistent current (I NaP). [•] This I NaP activates over a broad voltage range. [•] I260T contributes to I NaP at hyperpolarizing potentials via enhancement of window currents. [•] A1731V contributes to I NaP at depolarizing potentials probably by inhibiting the outward movement of IVS4. [ABSTRACT FROM AUTHOR]

Av3, a neurotoxin of Anemonia viridis, is toxic to crustaceans and cockroaches but inactive in mammals. In the present study, Av3 was expressed in Escherichia coli Origami B (DE3) and purified by reversed-phase liquid chromatography. The purified Av3 was injected into the hemocoel of Helicoverpa armigera, rendering the worm paralyzed. Then, Av3 was expressed alone or fusion expressed with the Cry1Ac in acrystalliferous strain CryB of Bacillus thuringiensis. The shape of Cry1Ac was changed by fusion with Av3. The expressed fusion protein, Cry1AcAv3, formed irregular rhombus- or crescent-shaped crystalline inclusions, which is quite different from the shape of original Cry1Ac crystals. The toxicity of Cry1Ac was improved by fused expression. Compared with original Cry1Ac expressed in CryB, the oral toxicity of Cry1AcAv3 to H. armigera was elevated about 2.6-fold. No toxicity was detected when Av3 was expressed in CryB alone. The present study confirmed that marine toxins could be used in bio-control and implied that fused expression with other insecticidal proteins could be an efficient way for their application. [ABSTRACT FROM AUTHOR]

The regulation of intracellular pH ( pHi) is a fundamental aspect of cell physiology that has received little attention in studies of the phylum Cnidaria, which includes ecologically important sea anemones and reef-building corals. Like all organisms, cnidarians must maintain pH homeostasis to counterbalance reductions in pHi, which can arise because of changes in either intrinsic or extrinsic parameters. Corals and sea anemones face natural daily changes in internal fluids, where the extracellular pH can range from 8.9 during the day to 7.4 at night. Furthermore, cnidarians are likely to experience future CO2-driven declines in seawater pH, a process known as ocean acidification. Here, we carried out the first mechanistic investigation to determine how cnidarian pHi regulation responds to decreases in extracellular and intracellular pH. Using the anemone Anemonia viridis, we employed confocal live cell imaging and a pH-sensitive dye to track the dynamics of pHi after intracellular acidosis induced by acute exposure to decreases in seawater pH and NH4Cl prepulses. The investigation was conducted on cells that contained intracellular symbiotic algae ( Symbiodinium sp.) and on symbiont-free endoderm cells. Experiments using inhibitors and Na+-free seawater indicate a potential role of Na+/H+ plasma membrane exchangers (NHEs) in mediating pHi recovery following intracellular acidosis in both cell types. We also measured the buffering capacity of cells, and obtained values between 20.8 and 43.8 m m per pH unit, which are comparable to those in other invertebrates. Our findings provide the first steps towards a better understanding of acid-base regulation in these basal metazoans, for which information on cell physiology is extremely limited. [ABSTRACT FROM AUTHOR]

Abstract: The present paper reports for the first time the bioaccessibility of arsenic species in Anemonia sulcata and its metabolization in the human body after the ingestion of this seafood, a delicacy highly appreciated in the south of Spain. Speciation in tissue extracts and human fluids was carried out by anion/cation exchange chromatography with inductively coupled plasma mass spectrometry ((IC)-ICP-MS). Results obtained show that about 54% of the arsenic present in the anemones is lost after cooking, around 95% of total arsenic in cooked anemones is bioaccessible and 85% of the amount ingested is eliminated from the body by urine in 90h. The relative abundance of As in urine varies along the time, but after the first 10h follows the order arsenobetaine (AB)≈DMAV (dimethylarsinate)>arsenocholine (AsC)>tetra-methyl-arsonium (TETRA)>trimethyl-arsine oxide (TMAO)>MAV (monomethylarsonate)>iAsV >dimethylarsenoethanol (DMAE), while the major species in cooked anemones is AB followed by DMAV >dimethylmonothioarsinic acid (DMASV)>AsC>TETRA>TMAO>iAsV >glycerylphosphorylarsenocholine(GPAsC)>MAV >DMA. After 30min from the ingestion, DMAV is the most abundant in human serum, followed by AB, MAV and AsC. Finally, the use of organic mass spectrometry allowed the standardless identification of AsC, GPAsC, DMASV and DMAE. [Copyright &y& Elsevier]

Abstract: Arsenic species have been investigated in Anemonia sulcata, which is frequently consumed food staple in Spain battered in wheat flour and fried with olive oil. Speciation in tissue extracts was carried out by anion/cation exchange chromatography with inductively coupled plasma mass spectrometry (HPLC-(AEC/CEC)–ICP-MS). Three methods for the extraction of arsenic species were investigated (ultrasonic bath, ultrasonic probe and focused microwave) and the optimal one was applied. Arsenic speciation was carried out in raw and cooked anemone and the dominant species are dimethylarsinic acid (DMAV) followed by arsenobetaine (AB), AsV, monomethylarsonic acid (MAV), tetramethylarsonium ion (TETRA) and trimethylarsine oxide (TMAO). In addition, arsenocholine (AsC), glyceryl phosphorylarsenocholine (GPAsC) and dimethylarsinothioic acid (DMAS) were identified by liquid chromatography coupled to triple quadrupole mass spectrometry (HPLC–MS). These results are interesting since GPAsC has been previously reported in marine organisms after experimental exposure to AsC, but not in natural samples. In addition, this paper reports for the first time the identification of DMAS in marine food. [Copyright &y& Elsevier]

Eur J Clin Invest 2012; Abstract Background Long QT syndrome (LQTS) is associated with a high incidence of atrial fibrillation (AF), but the underlying mechanisms are unclear. Pulmonary veins (PVs) play a critical role in AF genesis. Type 3 LQTS increases late sodium current (INa,L), which may increase PV arrhythmogenesis and AF. Therefore, this study examines PV arrhythmogenesis in anemonia sulcata toxin II (ATX-II)-induced type 3 LQTS and evaluates whether the INa,L inhibitor ranolazine can suppress PV arrhythmogenesis. Materials and methods Conventional microelectrodes were used to record the action potentials (AP) and contractility in isolated rabbit PV specimens before and after ATX-II administration with or without ranolazine. Results Anemonia sulcata toxin II (100 nM) increased the PV spontaneous rates from 2·0 ± 0·1 to 2·9 ± 0·2 Hz ( n = 7), induced PV burst firing (100%) with the genesis of early afterdepolarization (EAD) (86%) and prolonged the AP duration. Ranolazine (0·1, 1 and 10 μM) dose dependently reduced the PV spontaneous rates from 2·5 ± 0·2 to 2·3 ± 0·2 Hz, 1·9 ± 0·2 and 1·5 ± 0·3 Hz ( P < 0·05) and decreased the diastolic tension by 40 ± 19%, 87 ± 26% and 113 ± 28%. In the presence of ranolazine (10 μM), ATX-II (100 nM) further increased the AP duration. However, ATX-II neither increased the PV spontaneous rates (1·6 ± 0·1 vs. 1·7 ± 0·2 Hz, n = 7) nor induced PV burst firing or EAD. Moreover, ranolazine (10 μM) reduced ATX-II-induced PV acceleration and EAD. Conclusions The INa,L enhancer ATX-II can increase PV arrhythmogenesis, which can be attenuated or blocked by ranolazine. This suggests that AF may be related to type 3 LQTS through increased INa,L. [ABSTRACT FROM AUTHOR]

Abstract: A pathological increase in the late component of the cardiac Na+ current, INaL, has been linked to disease manifestation in inherited and acquired cardiac diseases including the long QT variant 3 (LQT3) syndrome and heart failure. Disruption in INaL leads to action potential prolongation, disruption of normal cellular repolarization, development of arrhythmia triggers, and propensity to ventricular arrhythmia. Attempts to treat arrhythmogenic sequelae from inherited and acquired syndromes pharmacologically with common Na+ channel blockers (e.g. flecainide, lidocaine, and amiodarone) have been largely unsuccessful. This is due to drug toxicity and the failure of most current drugs to discriminate between the peak current component, chiefly responsible for single cell excitability and propagation in coupled tissue, and the late component (INaL) of the Na+ current. Although small in magnitude as compared to the peak Na+ current (~1–3%), INaL alters action potential properties and increases Na+ loading in cardiac cells. With the increasing recognition that multiple cardiac pathological conditions share phenotypic manifestations of INaL upregulation, there has been renewed interest in specific pharmacological inhibition of INa. The novel antianginal agent ranolazine, which shows a marked selectivity for late versus peak Na+ current, may represent a novel drug archetype for targeted reduction of INaL. This article aims to review common pathophysiological mechanisms leading to enhanced INaL in LQT3 and heart failure as prototypical disease conditions. Also reviewed are promising therapeutic strategies tailored to alter the molecular mechanisms underlying INa mediated arrhythmia triggers. [Copyright &y& Elsevier]

In a marine envenomation, identification of the culprit organism can be difficult. In this case report, we present our method to identify snakelocks anemone ( Anemonia viridis or formerly Anemonia sulcata) as the culprit of a severe toxic skin reaction. A. viridis is one of the most common anemones of the Mediterranean Sea and the North Atlantic Ocean. It lives at a depth of up to 10 m. It is a member of the phylum Cnidaria, which includes jellyfish, anemones, hydroids, and corals. They have toxic organelles called cnidocysts that have the capacity to inject venom with microscopic harpoon-like structures. The cnidocysts of A. viridis may cause toxic and allergic reactions, and although its venom is one of the most studied cnidarian venoms, detailed case reports are rare. [ABSTRACT FROM AUTHOR]

The discharge and mode of action of the microbasic mastigophores (b-Rhabdoids) of Anemonia sulcata Pennant (Anthozoa) tentacles were examined using light and electron microscopical techniques. The statistical analysis of different morphological parameters of 100 nematocysts before and after their complete discharge revealed four different phases in the process of discharge. The first phase consists of the opening of the three opercular flaps. The second phase is characterized by the extrusion of the shaft. Osmotic pressure generated by the nematocyst propagates the extremely fast shaft discharge. The third phase, the evagination of the long tubule, is propagated by intrinsic mechanical forces stored in the tubule's twists and coils. During the process of evagination the tubule length increases by 95%, its circumference by 17%. Its forward motion is produced by the dilatation of its circumference and undoing of its twists. This motion is supported by the pressure which is generated by the hydration of the capsular matrix. The fourth phase consists in the extrusion of soluble capsular material at the tip of the evaginated tubule. During discharge the net volume of the nematocyst increases by 182%. This expansion is only possible if a high osmotic pressure is generated within the cyst. The extrusion of the shaft is the crucial phase in the discharge of the microbasic mastigophores, because without the puncture produced by the shaft the tubule is not able to overcome even surfaces of low viscosity media. The function of these tentacular nematocysts is the mechanical capture of prey organisms and, most probably, the introduction of unknown toxins into the latter. [ABSTRACT FROM AUTHOR]

Simple Summary: Colorectal cancer is one of the most frequent types of cancer in the population. Recently, invertebrate marine animals have been investigated for the presence of natural products which can damage tumor cells, prevent their spread to other tissues or avoid cancer develop. We analyzed the anemone Anemonia sulcata with and without the presence of its microalgal symbiont (Symbiodinium) as a source of bioactive molecules for the colorectal cancer therapy and prevention. Colon cancer tumor cells were exposed to Anemone extracts observing a remarkable cell death and a great antioxidant capacity. These preliminary results support that Anemonia sulcata could be a source of bioactive compounds against colorectal cancer and that the absence of its symbiont may enhance these properties. Further studies will be necessary to define the bioactive compounds of Anemonia sulcata and their mechanisms of action. Recently, invertebrate marine species have been investigated for the presence of natural products with antitumor activity. We analyzed the invertebrate Anemonia sulcata with (W) and without (W/O) the presence of its microalgal symbiont Symbiodinium as a source of bioactive compounds that may be applied in the therapy and/or prevention of colorectal cancer (CRC). Animals were mechanically homogenized and subjected to ethanolic extraction. The proximate composition and fatty acid profile were determined. In addition, an in vitro digestion was performed to study the potentially dialyzable fraction. The antioxidant and antitumor activity of the samples and the digestion products were analyzed in CRC cells in vitro. Our results show a high concentration of polyunsaturated fatty acid in the anemone and a great antioxidant capacity, which demonstrated the ability to prevent cell death and a high antitumor activity of the crude homogenates against CRC cells and multicellular tumor spheroids, especially W/O symbiont. These preliminary results support that Anemonia sulcata could be a source of bioactive compounds with antioxidant and antitumor potential against CRC and that the absence of its symbiont may enhance these properties. Further studies will be necessary to define the bioactive compounds of Anemonia sulcata and their mechanisms of action. [ABSTRACT FROM AUTHOR]

Marine-derived fungi are an important source of many valuable compounds with original structures and diverse physico-chemical properties. In this work, the metabolomic profile of a strain of Penicillium brevicompactum, recovered from a snakelocks sea anemone (Anemonia sulcata), was investigated through the parallel application of LC-ESI-HRMS, GC-MS, and NMR. Our strategy allowed the identification of mycophenolic acid, brevianamide A, and several compounds belonging to the thiosilvatins. Among the latter, five products are reported for the first time in this species. The main product of this series, cis-bis(methylthio)silvatin, was also tested for antiproliferative activity on both cancer and non-tumoral colon cell lines. [ABSTRACT FROM AUTHOR]