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Histone deacetylases (HDACs) are key components of the epigenetic machinery controlling gene expression. They are involved in chromatin remodeling events via post-translational histone modifications but may also act on nonhistone proteins, influencing many fundamental cellular processes. Due to the key involvement of HDACs in serious human pathologies, including cancer, HDAC inhibitors (HDACis) have received increased attention in recent years. It is known that marine invertebrates produce significant amounts of secondary metabolites showing active pharmacological properties and an extensive spectrum of biomedical applications. Some of these compounds possess HDACi properties.

Marine pollution has been increasing as a consequence of human activities. In the near future will start the extraction of minerals from ocean depths, Deep Sea Mining (DSM). However, the impacts at the ecosystem level are still unknown. It is necessary to know the effects to contribute to the creation of rules, laws and mining codes that protect and conserve ecosystems and biodiversity. Sea urchin and mussels represents one of the most important seafood product, but also, a sentinel organism for ecotoxicological studies to assess aquatic environments quality. They are directly exposed to anthropogenic contaminants in their habitat. The aim of this study is to understand the influences of two trace element (which will probably be released into the water column by DSM) at three different environmentally realistic concentrations in Arbacia lixula and Mytilus galloprovincialis: Cu (0.6μg/l; 339.15 μg/l; 1709 μg/l), and Pb (1.0 μg/l; 619.16 μg/l; 2291 μg/l). We observed the short-term (3 and 7 days) effects at biochemical level (total cells count, cell mortality, total protein concentration and lysosomal membrane stability) and their bioaccumulation in different tissues. In A. lixula the bioaccumulation was studied in gonads while in M. galloprovincialis in gills and mantle. This study demonstrates the importance of biomonitoring contaminants and understanding how biomarkers relate to each other on exposure to particular contaminants in different species due to their wide application in environmental management. Few studies have used multiple biomarkers in more than a single species to learn about the variability of the sub-lethal effects of a contaminant

The drugs, agents for diseases therapy and prevention, are ubiquitous environmental pollutants. We analyzed a specific anti-inflammatory, diclofenac, present in soil and water. The study focused on Arbacia lixula (Linnaeus, 1758) and Paracentrotus lividus (Lamarck, 1816) reproductive success. In A. lixula we identified the drug qualitative and quantitative fertilization effects, and if they affect the functional male and female gametes capacity; in P. lividus we analyzed the drug effects and the male and female gametes quantitative integrity. Through the High Liquid Chromatografy we quantitatively evaluated the real diclofenac contribution on A. lixula embryos and on A. lixula and P. lividus reproductive cells. The results confirmed that the diclofenac is an agent that influence the A. lixula fertilization. The embryos shown: morphological aberrations and slowdown of embryonic development with delay cell divisions. The A. lixula gametes, treated with five toxic solutions, showed modifications or alterations: the eggs are lysed and the spermatozoa shown a decrease in sperm speed. In P. lividus male gametes we observed a real decrease in sperm speed but the egg cells, apparently, no have morpho-structural change although it is found a real diclofenac absorption. The study finds that the two species have a diclofenac sensitivity and that the male gametes are more vulnerable and sensitive to high diclofenac concentrations compared to female gametes. Much remains to be done to increase the knowledges despite today we known the potential risks due to the drugs presence in the waters and being that many of these substances have additive or synergistic activity and their effect can be greatly enhanced.

OBJECTIVES: Thrombocytopenia is a hallmark for patients with cirrhosis and it is perceived as a risk factor for bleeding events. However, the relationship between platelet count and bleeding is still unclear. METHODS: We investigated the relationship between platelet count and major or clinical relevant nonmajor bleedings during a follow-up of ∼4 years. RESULTS: A total of 280 cirrhotic patients with different degrees of liver disease (67% males; age 64±37 years; 47% Child–Pugh B and C) were followed up for a median of 1,129 (interquartile range: 800–1,498) days yielding 953.12 patient-year of observation. The annual rate of any significant bleeding was 5.45%/year (3.57%/year and 1.89%/year for major and minor bleeding, respectively). Fifty-two (18.6%) patients experienced a major (n=34) or minor (n=18) bleeding event, predominantly from gastrointestinal origin. Platelet counts progressively decreased with the worsening of liver disease and were similar in patients with or without major or minor bleeding: a platelet count ≤50×103/μl was detected in 3 (6%) patients with and in 20 (9%) patients without any bleeding event. Conversely, prothrombin time-international normalized ratio was slightly higher in patients with overall or major bleeding. On Cox proportional hazard analysis, only a previous gastrointestinal bleeding (hazard ratio (HR): 1.96; 95% confidence interval: 1.11–3.47; P=0.020) and encephalopathy (HR: 2.05; 95% confidence interval: 1.16–3.62; P=0.013) independently predicted overall bleeding events. CONCLUSIONS: Platelet count does not predict unprovoked major or minor bleeding in cirrhotic patients.

Phylogenetic analysis recognizes echinoderms as a key group of deuterostomes, therefore the species in this group are useful for the study of the evolution of innate immunity responses. In addition, this marine invertebrate lives in coastal and estuarine waters that are directly exposed to potentially pathogenic microorganisms and stressful anthropogenic factors. Coelomocyte populations seem to be essential to immune-defence functions such as phagocytosis, ROI production, cytotoxicity, synthesis and release of antimicrobial substances including lectin, cytokine, C3-like expression, prophenoloxidase activity and capsule formation. Holothurians’ coelomocyte populations contain several coelomocyte types, including phagocytes, and can form brown bodies in response to multicellular parasites. Holothuria tubulosa coelomic fluid contains three main coelomocyte categories: amoebocytes, spherulocytes and progenitor cells. The amoebocytes represent about 30 % of the total population, and in living cell preparations exhibit two distinct forms: petaloid and philopodial; spherule cells represent the numerically dominant cell type (about 60 %). Progenitor cells are present in a lesser amount (about 20 %); they are similar to lymphocytes and show a nucleus that is typically prominent with a thin rim of cytoplasm. In the present report we show that coelomocytes of Holothuria tubulosa are able to exert cytotoxic activity against different cellular targets such as rabbit or sheep erythrocytes and the human erythromyeloid leukaemia-derived cell line K562. Moreover, the unseparated coelomocyte supernatant lysate (CLS) exerts a lytic activity, even in the absence of calcium, against the same targets. Analysis of the coelomocyte lysate by overlay assays with sheep and rabbit erythrocytes on PAGE without SDS showed a protein pattern composed by two main hemolytic bands with different electrophoretic mobility. The one with low mobility (I) showed calcium independent haemolytic activity while high mobility band (II) showed calcium dependent activity. The two bands were eluted from the gel and analyzed by SDS-PAGE and stained with silver stain; band I could be separated in three components of different sizes (52, 42 and 41 kDa), and band II had a size of 43 kDa. Spherule cells seemed to be the effector cells, as shown by a plaque lysis assay. Further studies are in progress to identify the lytic proteins and the lytic mechanism at work on both a molecular and cellular level.

Porphyrins are an important class of natural and artificial pigments which play an important role in largely different area of both fundamental and technological interest. In particular, porphyrin metal derivatives have been exploited as models for enzymes and artificial blood. Charged porphyrins are able to interact with several relevant biomolecules, i.e., nucleic acids, polypeptides, and proteins. This property together with their ability to localize into tumor cells and to photosensitize the production of singlet oxygen led to the development of several compounds actually in use or under investigation for photodynamic therapy applications (PDT) [1]. Interestingly, quite recently, meso-tetrakis(4-carboxyphenyl) porphyrin (TPPC) and its derivatives have been exploited as a marker for the rapid detection of tumor cells by fluorescence imaging. A common feature of these molecules is their propensity to interact to form dimers, oligomers, or more extended aggregates. To develop efficient systems for biomedical applications or for PDT (in which aggregation should be prevented) or to stabilize monomeric porphyrins in a very water-soluble form, novel systems based on biocompatible delivery systems are highly desirable. In this framework, recently some of us reported on the employment of biocompatible amino-terminated polypropylene or poly(ethylene oxide)s generally termed as Jeffamines, to prevent porphyrin aggregation, allowing to reach millimolar concentration of TPPC in a monomeric form in solution [2]. In biological media, cell membranes seem to be important targets for many antineoplastic photosensitizer agents. Red blood cells have been often used for in vitro PDT studies. Here we report on two different porphyrin derivatives, TPPC-Jeff and ZnTPPC-Jeff. Photodynamic action was then evaluated in vitro using human red blood (HRB) cells under different conditions to obtain information about the effect produced by these porphyrin derivatives. The morphology of erythrocytes has been investigated by optical microscopy after incubation with porphyrin compounds and irradiation having a losing of their normal biconcave profile and an incoming of a spiny configuration with blebs in their surfaces. Further studies are required to optimize potential therapeutic dosing strategies to inform and encourage clinical trial design.

Spectroscopic characterization and in vitro assay on human blood of novel porphyrin derivatives

Aqueous extracts from Posidonia oceanica’s green and brown (beached) leaves and rhizomes were prepared, submitted to phenolic compound and proteomic analysis, and examined for their potential cytotoxic effect on HepG2 liver cancer cells in culture. The chosen endpoints related to survival and death were cell viability and locomotory behavior, cell-cycle analysis, apoptosis and autophagy, mitochondrial membrane polarization, and cell redox state. Here, we show that 24 h exposure to both green-leaf- and rhizome-derived extracts decreased tumor cell number in a dose–response manner, with a mean half maximal inhibitory concentration (IC50) estimated at 83 and 11.5 μg of dry extract/mL, respectively. Exposure to the IC50 of the extracts appeared to inhibit cell motility and long-term cell replicating capacity, with a more pronounced effect exerted by the rhizome-derived preparation. The underlying death-promoting mechanisms identified involved the down-regulation of autophagy, the onset of apoptosis, the decrease in the generation of reactive oxygen species, and the dissipation of mitochondrial transmembrane potential, although, at the molecular level, the two extracts appeared to elicit partially differentiating effects, conceivably due to their diverse composition. In conclusion, P. oceanica extracts merit further investigation to develop novel promising prevention and/or treatment agents, as well as beneficial supplements for the formulation of functional foods and food-packaging material with antioxidant and anticancer properties.

Introduction: Echinoderm coelomocytes have traditionally been investigated through a morphological approach using light microscopy, which relies on the idea of constant cell shape as a stable character. However, this can be affected by biotic or abiotic conditions. Objective: To analyze if the consistency in cell morphology offered by the cytocentrifugation method, might be used as a convenient tool to study echinoderm coelomocytes. Methods: Cells of Echinaster (Othilia) brasiliensis (Asteroidea), Holothuria (Holothuria) tubulosa (Holothuroidea), Eucidaris tribuloides, Arbacia lixula, Lytechinus variegatus, and Echinometra lucunter (Echinoidea) were spread on microscope slides by cytocentrifugation, stained, and analyzed through light microscopy. Additionally, fluorescence microscopy, scanning electron microscopy, and energy-dispersive x-ray spectroscopy were applied to cytospin preparations, to complement the analysis of granular and colorless spherulocytes of Eucidaris tribuloides. Results: Altogether, 11 cell types, including phagocytes, spherulocytes, vibratile cells, and progenitor cells were identified in the samples analyzed. The granular spherulocyte, a newly-described cell type, was observed in all Echinoidea and was very similar to the acidophilic spherulocytes of Holothuria (Holothuria) tubulosa. Conclusions: Cytocentrifugation proved to be versatile, either as the main method of investigation in stained preparations, or as a framework on which other procedures may be performed. Its ability to maintain a constant morphology allowed accurate correspondence between live and fixed/stained cells, differentiation among similar spherulocytes as well as comparisons between similar cells of Holothuroidea and Echinoidea. Resumen Introducción: Los celomocitos de equinodermos se han investigado tradicionalmente a través de un enfoque morfológico utilizando microscopía óptica, que se basa en la idea de la forma celular constante como un carácter estable. Sin embargo, esto puede verse afectado por condiciones bióticas o abióticas. Objetivo: Analizar si la consistencia en la morfología celular que ofrece el método de citocentrifugación podría utilizarse como una herramienta conveniente para estudiar los celomocitos de equinodermos. Métodos: Células de Echinaster (Othilia) brasiliensis (Asteroidea), Holothuria (Holothuria) tubulosa (Holothuroidea), Eucidaris tribuloides, Arbacia lixula, Lytechinus variegatus y Echinometra lucunter (Echinoidea) se esparcieron en portaobjetos de microscopio por citocentrifugación, se tiñeron y analizaron mediante microscopía óptica. Adicionalmente, se aplicó microscopía de fluorescencia, microscopía electrónica de barrido y espectroscopía de rayos X con dispersión de energía a las preparaciones de citoespina, para complementar el análisis de los esferulocitos granulares e incoloros de Eucidaris tribuloides. Resultados: En total, se identificaron en las muestras analizadas 11 tipos de células, incluidos fagocitos, esferulocitos, células vibrátiles y células progenitoras. El esferulocito granular, un tipo de célula recién descrito, se observó en todos los Echinoidea y fue muy similar a los esferulocitos acidófilos de Holothuria (Holothuria) tubulosa. Conclusiones: La citocentrifugación demostró ser un método bastante versátil, ya sea como el método principal de investigación en preparaciones teñidas o como un marco en el que se pueden realizar otros procedimientos. Su capacidad para mantener una morfología constante permitió una correspondencia precisa entre las células vivas y las células fijas/teñidas, la diferenciación entre esferulocitos similares, así como comparaciones entre células similares de Holothuroidea y Echinoidea.

Territoriality, spatial exploration and social hierarchy are strictly related behaviors in gregarious fishes, and are often under-appreciated in farms where the individuals are confined within crowded spaces. In this study, we investigated the role of spatial exploration, elucidating the importance of time upon forming the social organization, and the role of the territoriality in gilthead seabream (Sparus aurata), using two experimental approaches. In the first approach, three fish were placed sequentially in the aquarium with an interval of two days (sequential model), while in the second (simultaneous model), two fish were simultaneously placed in an aquarium divided by a barrier which was removed after a certain period of time. To study the effect of social stress and spatial perception in the two models, we monitored behavior (aggressive acts and feeding priority), integrated with the evaluation of physiological and cellular stress parameters, such as phagocytosis, cortisol, glucose, and blood osmolarity levels. After the establishment of the social hierarchy in the “sequential model”, we observed that the levels of cortisol and an immunological cell-mediated marker were higher in subordinate individuals than in the dominant ones. We observed a different modulation of phagocytic activity in peritoneal cavity cells between dominant and subordinates, demonstrating that social stress acts upon immune response. Differently from the first model, no behavioral, physiological, or phagocytic differences were found between the two fish involved in the simultaneous model, where both fish acted as co-dominants, defending their territory. The study achieved a deeper understanding of the role of spatial exploration, territorial dominance and intraspecific interaction in gilthead seabream, and elucidated the link between them and physiological stress indicators. The results highlight aspects of interest to the aquaculture industry, showing the importance of a greater focus on rearing conditions, finding solutions to mitigate crowding effects and promoting the quality of aquacultural products.

Recently described biochemical and structural aspects of fucose-binding lectins from the European eel (Anguilla anguilla) and striped bass (Morone saxatilis) led to the identification of a novel lectin family ("F-type" lectins) characterized by a unique sequence motif and a characteristic structural fold. The F-type fold is shared not only with other members of this lectin family, but also with apparently unrelated proteins ranging from prokaryotes to vertebrates. Here we describe the purification, biochemical and molecular properties, and the opsonic activity of an F-type lectin (DlFBL) isolated from sea bass (Dicentrarchus labrax) serum. DlFBL exhibits two tandemly arranged carbohydrate-recognition domains that display the F-type sequence motif. In situ hybridization and immunohistochemical analysis revealed that DlFBL is specifically expressed and localized in hepatocytes and intestinal cells. Exposure of formalin-killed Escherichia coli to DlFBL enhanced their phagocytosis by D. labrax peritoneal macrophages relative to the unexposed controls, suggesting that DlFBL may function as an opsonin in plasma and intestinal mucus.

The interest in the consumption of health-promoting foods has led to identifying derivatives of the wine industry as products to increase the functional properties of different foods or to design new functional foods. The main goal of this study is to characterize and valorize byproducts and wastes of Sicilian grapes as new sources of bioactive components, from the perspective of a circular economy and a biorefinery approach. In particular, this research investigated: 1. the total phenolic content and antioxidant activities and 2. the phenolic profiles of free and bound fractions of defatted grape seeds and red grape skins from Sicily. Defatted grape seeds (DGS) and red grape skins (RGSK) are rich in phenolic compounds. Twenty biophenols were found in the defatted seeds and red grape skins. Particularly interesting were the results obtained after basic hydrolysis, which allowed the release of biophenols from the matrix. The degreased grape seeds showed p-coumaric acid levels at 4641.65 µg g−1, gallic acid at 2649.23 µg g−1, and caffeic acid at 1474.13 µg g−1, along with appreciable quantities of myricetin, epicatechin, and quercetin. As a sustainable approach, the reuse and the value added of the byproducts and wastes of grapes grown in Sicily is shown, which makes possible new applications in different fields, i.e., nutraceuticals.

Coelomocytes in the Holothuroidea are traditionally identified according to their morphology through light, fluorescence, or electron microscopy. Former studies have typically used only one method, with few works combining two or more approaches. Studies using cytocentrifugation to study these cells are scarcer. Thus, for the first time, an integrative approach was used to compare coelomocytes in Holothuroidea. This approach consisted of living and stained cells, scanning electron microscopy (for spherule cells), and accurate morphometric analyses. Specifically, we used specimens of Holothuria grisea, Holothuriaarenicola, and Holothuriatubulosa to test whether cytocentrifugation could be valuable in comparative studies with coelomocytes, whether an integrative approach could help to understand spherule cell diversity, and whether closely related species, even those having distinct geographic distributions and ecological requirements, would have a similar population of coelomocytes. Our results showed seven distinct cell types in these species, including phagocytes, fusiform cells, morula cells, acidophilic spherulocytes, spherulocytes, progenitor cells, and crystal cells. Total and differential cell counts, along with morphometric parameters, were similar among species. Morphometric analyses of spherule cells revealed consistent differences among the diameter of their cytoplasmic spherules, as well as a set of different morphotypes in acidophilic spherulocytes and spherulocytes. Cytospin preparations proved to be quite useful because they provided constant morphological and morphometric data, allowing accurate identification of the cell types and comparisons among species. Moreover, this study highlighted (1) that the spherule diameter is a good parameter to separate spherule cells and (2) a putative maturation process to acidophilic spherulocytes and spherulocytes. Lastly, we showed that the cells of these species are very similar, regardless of their geographic distribution and ecology. Thus, our work contributes to a better understanding of the coelomocytes in Holothuria, a genus with a wide geographic distribution. The present study may be useful to establish these species as important model organisms, as well as bring insights into the functions of coelomocytes.

Among the Sicilian economic productive sectors, that of wine production has today a considerable economic value. However, with the growth of this sector, notable was the increase in the production of waste, which to date is not only an economic damage for companies, but also a threat to the environment. It is known that waste from wine production has properties (e.g., antioxidants) which have potential reuse at cosmetic, pharmaceutical and nutritional levels to obtain economically sustainable applications. A new goal is given by the recovery of added value compounds from agri-food wastes and by-products. Grape seed oil is a promising vegetable fat and cold pressing does not involve the use of chemicals, which are harmful to health. It implies that cold-pressed seed oils may contain phytochemicals, as well as natural antioxidants, more than refined oils. In this context, this works aims at studying the chemical characterization (triglycerides profile and composition in fatty acids) of grape seed oils obtained from Soxhlet and cold pressed extraction from Sicilian red grape seeds and white grape seeds. The possibility of obtaining high yields of triglycerides and fatty acids from the waste of wine production through new extraction methods would open up new perspectives for the reuse of waste in a human and animal food context. The results of this work allow the opening up of new perspectives to reuse and then reduce these wastes, helping not only to reduce the damage to the environment and costs for companies but also to create a new product that is environmentally sustainable and with an important economic value.