In Vitro Antioxidant Activity and In Vivo Topical Efficacy of Lipid Nanoparticles Co-Loading Idebenone and Tocopheryl Acetate

Idebenone (IDE) is a strong antioxidant that has been proposed for the treatment of skin disorders, including skin ageing. Unfavorable physico-chemical properties make IDE a poor skin permeant where effectiveness could be improved by its loading into suitable delivery systems such as solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC). In this work, we designed novel IDE-loaded NLC containing tocopheryl acetate (VitE) as a liquid component to obtain a synergic effect between IDE and VitE. The resulting NLC showed small particle sizes (24-42 nm), low polydispersity indices (<0.300), good stability, and were assessed for their in vitro antioxidant activity and in vivo topical effects. IDE-loaded SLN and NLC showed a high antioxidant activity in in vitro assays (DPPH and reducing power method) and provided a similar and significant protection from oxidative stress of fibroblast cells, HS-68, exposed to UV light. After a two-week topical treatment of human volunteers with gels containing IDE-loaded SLN or NLC, a similar increase in skin hydration was observed, while IDE NLC reduced skin pigmentation to a greater extent than IDE SLN. These results suggest that co-loading IDE and VitE into NLC could be a promising strategy to obtain topical formulations with improved photo-protection

Dietary Inclusion of Halobacterium salinarum Modulates Growth Performances and Immune Responses in Farmed Gilthead Seabream (Sparus aurata L.)

The use of natural immunostimulants is considered the most promising alternative to promote fish health, productive performance and quality, increasing the aquaculture profitability, sustainability and social acceptance. The purpose of this study was to evaluate the effect of the integration of a potential probiotic strain, Halobacterium salinarum, belonging to the Archaea domain, in the formulated diets of farmed gilthead seabream (Sparus aurata L.) in terms of growth performances and immunity responses. The experiment was set up to test two different levels of inclusion of the bacteria in the diet: 0.05% (D1) and 0.1% (D2). The effects on fish growth performances; humoral (peroxidase, protease, antiprotease and IgM levels) and cellular immunity parameters (phagocytosis, respiratory burst and myeloperoxidase), along with bactericidal activity, were evaluated after 15 and 30 days of experimental feeding. The obtained results showed that the inclusion of H. salinarum at the highest concentration (D2 0.1%) improved growth performances, bactericidal activity against Vibrio anguillarum and some parameters related both to the humoral and cellular immune response, suggesting exploring other aspects of welfare in view of future supplementations of this probiotic strain in the diet of S. aurata

Effects of Dietary Supplementation with Honeybee Pollen and Its Supercritical Fluid Extract on Immune Response and Fillet’s Quality of Farmed Gilthead Seabream (Sparus aurata)

The awareness of the correlation between administered diet, fish health and products’ quality has led to the increase in the research for innovative and functional feed ingredients. Herein, a plant-derived product rich in bioactive compounds, such as honeybee pollen (HBP), was includ-ed as raw (HBP) and as Supercritical Fluid Extracted (SFE) pollen (HBP_SFE) in the diet for gilt-head seabream (Sparus aurata). The experiment was carried out on 90 fish with an average body weight of 294.7 ± 12.8 g, divided into five groups, according to the administration of five diets for 30 days: control diet (CTR); two diets containing HBP at 5% (P5) and at 10% (P10) level of in-clusion; two diets containing HBP_SFE, at 0.5% (E0.5) and at 1% (E1) level of inclusion. Their ef-fects were evaluated on 60 specimens (336.2 ± 11.4 g average final body weight) considering the fish growth, the expression of some hepatic genes involved in the inflammatory response (il-1β, il-6 and il-8) through quantitative real-time PCR, and physico-chemical characterization (namely color, texture, water holding capacity, fatty acid profile and lipid peroxidation) of the fish fillets monitored at the beginning (day 0) and after 110 days of storage at −20 °C. The results obtained showed that the treatment with diet E1 determined the up-regulation of il-1β, il-6, and il-8 (p < 0.05); however, this supplementation did not significantly contribute to limiting the oxidative stress. Nevertheless, no detrimental effect on color and the other physical characteristics was observed. These results suggest that a low level of HBP_SFE could be potentially utilized in aq-uaculture as an immunostimulant more than an antioxidant, but further investigation is neces-sary

Fisheries and aquaculture by-products modulate growth, body composition, and omega-3 polyunsaturated fatty acid content in black soldier fly (Hermetia illucens) larvae

The black soldier fly (BSF) (Hermetia illucens L.) is one of the most promising species for the production of ingredients, mainly protein, useful for animal feed formulation, owing to its ability to convert organic waste into biomass with a high nutritional value. However, the low percentage of n-3 series polyunsaturated fatty acids (PUFAs) in its fatty acid profile is a limiting factor for the utilization of BSF in fish feed. Recent studies have highlighted that wastes derived from different agro-food value chains could affect the nutritional composition of BSF larvae, depending on the composition of the wastes used as larvae-growing substrate. Due to the significant amount of n-3 PUFA in marine products, both in edible produce and in by-products, in this study, different sources of fish by-products were included in experimental diets for BSF to evaluate the effect of its addition on the final nutritional profile of BSF pre-pupae. One control diet and five experimental diets were prepared to feed the BSF larvae: wheat bran as the control diet (diet B), bycatch from Mediterranean trawl fisheries (diet F), Parapenaeus longirostris processing by-products (diet S), aquaculture processing by-products (diet R), Thunnus albacares processing by-products (diet T), and Engraulis encrasicolus processing by-products (diet A). In this study, the effects of the different diets were analyzed on the growth, body composition, and fatty acid profile of BSF larvae and pre-pupae. The obtained results showed that the different experimental diets affected total lipids content and fatty acids composition, when compared with the control. A significant increase in eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) in BSF larvae and pre-pupae fed with all fish by-products was observed when compared with those fed with diet B, in particular in larvae and pre-pupae fed with diet A, demonstrating that the utilization of fish processing by-products is a suitable solution for improving the nutritional value of insects as ingredients in aqua feeds. The reuse of marine by-products can contribute to the industry’s “zero waste” goal, increasing the sustainability of the fishery value chain and the formulation of new valuable products

Sub-lethal Doses of Polybrominated Diphenyl Ethers, in Vitro, Promote Oxidative Stress and Modulate Molecular Markers Related to Cell Cycle, Antioxidant Balance and Cellular Energy Management

In the present study, we evaluated the effects of different concentrations of the polybrominated diphenyl ethers (PBDEs) BDE-209, BDE-47 and BDE-99, on the vitality and oxidative stress of a HS-68 human cell culture exposed to the compounds for three days. The results showed that for this exposure time, only the highest concentrations produced a significant vitality reduction and oxidative stress induction (p < 0.05), measured as reactive oxygen species (ROS). Subsequently, in order to verify the effects of sub-lethal doses, cells were exposed for a longer time and data collected, after 12 and 20 days, to study ROS production and some molecular markers related to cell cycle and stress (p53, pRB, PARP, c-Jun and c-Fos), antioxidant status and proliferation (ERK, c-Jun and c-Fos), energy balance (NRF2, AMPK, HIF). Most of the biomarkers were influenced by the treatments, indicating that sub-lethal doses of PBDEs, for longer time, can enhance the production of ROS, altering the energetic metabolism, cell cycle and antioxidant balance, determining possible negative effects on the cell proliferation equilibrium

Valorization of low value fisheries species in Sicily (Italy), by realization of functional food: a case study of high technological readiness level

The growing demand for fish products of safe and local origin and the need to reduce fishing on target species, requires continuous diversification in the seafood industry, to meet the needs of consumers. One solution is represented by the use of surplus species of local fisheries, of little commercial value and often ignored by the consumer, which, if subjected to new preservation and processing methodologies, allow to create new product categories, which open up new market opportunities. In our study, we setup and applied, on some local surplus fisheries species from western Sicily, some processing technologies, aimed to valorize and extend the shelf life of fish products: the cold smoking with addition of natural antioxidants and the salting-drying, with low salt content. The cold smoking process was applied on fillets of the species Coryphaena hippurus, a surplus seasonal species. The process demonstrated to be effective in extend the shelf-life and ameliorate some sensorial aspects of quality. The salting-drying process was applied on the species Spicara smaris, after the setup of a protocol aimed to reduce the sodium content of the processed product. For this purpose, the sodium chloride was partially replaced with potassium chloride, during the salting phase. The partial replacement seems to be the best way to reduce the sodium content; both salts, in fact, have similar properties, the potassium, in addition, does not seem to be correlated with the onset of hypertension and cardiovascular diseases like the sodium. This allows to obtain a product which can be considered a functional food, intended also for consumers that suffer of hypertension. Both processes are in advanced phase of technological readiness level (TRL8) since have been tested also in a real environment through involvement of producers and consumers in the research project. Projects: “Biotechnologie Marine vecteur d’innovation et qualité” BioVecQ IEVP-2007–2013 Cod PS1.3_08; Strategic Use of Competitiveness towards Consolidating the Economic -Sustainability of the european Seafood Sector SUCCESS- HORIZON 2020-Grant n° 63518

Biotechnological Applications for the Sustainable Use of Marine By-products: In Vitro Antioxidant and Pro-apoptotic Effects of Astaxanthin Extracted with Supercritical CO2 from Parapeneus longirostris

In this study, the carotenoid astaxanthin was obtained by supercritical fluid extraction (SFE) from shrimp by-products (SBP). Its bioactive properties were evaluated in vitro in human normal and cancerous cells lines. The antioxidant activity of the extracted astaxanthin of the SFE fraction (ASTA) was tested in fibroblast cells (HS-68), by inducing oxidative stress and by evaluating the protective effect of the pre-treatment with different levels of ASTA against toxicity. The anti-proliferative activity was evaluated in hepatoma cells (HEP-G2), treated with increased concentrations of ASTA and measuring the effects on vitality and on some biomolecular markers related to oxidative stress, cell cycle, and apoptosis. It was found that pre-treating normal fibroblast cells with ASTA resulted in a marked increase in cell viability in a dose-dependent manner (P < 0.05) attesting its antioxidant power; in cancer cell line, increased concentrations of ASTA caused a time-dose-dependent decrease in the vitality, attesting its anti-proliferative activity (P < 0.05). The increased levels of the protein p-53 and the reduced levels of the proteins c-Jun and c-Fos at higher concentrations of ASTA, as well as, suggest the pro-apoptotic and anti-cancerous effects that this extract has on hepatocellular carcinomas, confirmed also by caspase-3 activation. These findings suggest biotechnological utilisation of marine by-products for nutraceutical and pharmaceutical applications avoiding the employment of organic solvents for extraction

Carbamazepine, cadmium chloride and polybrominated diphenyl ether-47, synergistically modulate the expression of antioxidants and cell cycle biomarkers, in the marine fish cell line SAF-1

A wide range of contaminants, industrial by-products, plastics, and pharmaceutics belonging to various categories, have been found in sea water. Although these compounds are detected at concentrations that might be considered as sub-lethal, under certain conditions they could act synergistically producing unexpected effects in term of toxicity or perturbation of biochemical markers leading to standard pathway. In this study, the Sparus aurata fibroblast cell line SAF-1, was exposed to increasing concentrations of carbamazepine (CBZ), polybrominated diphenyl ether 47 (BDE-47) and cadmium chloride (CdCl2) until 72 h, to evaluate the cytotoxicity and the expression of genes related to antioxidant defense, cell cycle and energetic balance. In general, both vitality and gene expression were affected by the exposure to the different toxicants, in terms of antioxidant defense and cell cycle control, showing the most significant effects in cells exposed to the mixture of the three compounds, respect to the single compounds separately. The synergic effect of the compounds on the analyzed biomarkers, underlie the potential negative impact of the contaminants on health of marine organisms

Oxidative Stress, Induced by Sub-Lethal Doses of BDE 209, Promotes Energy Management and Cell Cycle Modulation in the Marine Fish Cell Line SAF-1

The effects of sub-lethal doses of polybrominated diphenyl ether (PBDE)-209 in terms of toxicity, oxidative stress, and biomarkers were evaluated in the Sparus aurata fibroblast cell line (SAF-1). Vitality and oxidative stress status were studied after incubation with PBDE for 72 h. Concomitantly, the quantification of proteins related to cell cycle and DNA repair (p53), cell proliferation (extracellular signal⁻regulated kinase 1 (ERK1)), energetic restriction (hypoxia-inducible factor 1 (HIF1)), and redox status (Nuclear factor erythroid 2⁻related factor 2 (NRF2)) was also determined after prolonged exposure (7⁻15 days) by immunoblotting. Our results demonstrated that rising concentrations of PBDEs exposure-induced oxidative stress, and that this event modulates different cell pathways related to cell cycle, cell signaling, and energetic balance in the long term, indicating the negative impact of sub-lethal dose exposure to cell homeostasis