Background: Photodynamic therapy (PDT) is a cancer treatment that combines a photosensitizer (PS), light, and oxygen to generate reactive oxygen species (ROS), leading to tumor cell death. PDT efficacy depends largely on PS accumulation within tumors, prompting the development of third-generation PSs and nanotechnology-based delivery systems. Among these, BODIPYs (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) are promising PSs due to their favorable photophysical properties, while liposomes improve drug delivery, cellular uptake, and sustained release profiles. This study describes the synthesis of two novel BODIPY derivatives differing in the position of a methyl ester group on the meso-phenyl ring, their incorporation into liposomes, and evaluation of PDT efficacy. Methods: Cellular uptake of BODIPY-loaded liposomes, intracellular ROS generation, apoptosis, necrosis, and lipid peroxidation were assessed by flow cytometry in colorectal and ovarian cancer cell lines. The antitumor activity of the liposomal formulations was further evaluated in both 2D and 3D models using MTT and clonogenic assays. The involvement of ferroptosis and necroptosis in PDT-induced cell death was also investigated. Results: Liposomal formulations significantly enhanced cellular uptake compared with free compounds. Following light activation, both formulations induced potent antitumor effects through multiple cell death mechanisms, including canonical and non-canonical pathways, and maintained strong efficacy in 3D tumor spheroids. Conclusions: Liposome-encapsulated BODIPYs represent promising PDT agents by improving cellular uptake and eliciting robust antitumor activity through complementary cell death mechanisms. Furthermore, the methyl ester substituent on the meso-phenyl ring provides a versatile platform for future conjugation with targeting ligands, supporting the development of third-generation, tumor-targeted photosensitizers and warranting further preclinical investigation.

Shrimp by-products are a promising source of bioactives. This study investigated the composition and the bioactivity of a preparation obtained in mild conditions via ethanol extraction from the cephalothorax of the Mediterranean red shrimp species Aristaeomorpha foliacea. The crude extract (CE) exhibited high levels of astaxanthin (AST) and polyphenols, which contributed to its notable antioxidant activity. Monounsaturated fatty acids dominated the lipid profile of CE (46%), reflecting a typical trait of deep-sea species, while polyunsaturated and saturated fatty acids accounted for 33% and 21%, respectively. Chemical fractionation yielded a hydrophobic fraction (HF; AST-enriched) and a polar fraction (PF; polyphenols-enriched). In hepatic (HuH7) and intestinal (HCT15) human cells, CE induced dose-dependent lipid droplet accumulation and protected against palmitic acid-induced lipotoxicity: it restored cell viability, reduced reactive oxygen species, prevented lipid peroxidation and induced modulation of antioxidant enzymes, including catalase, glutathione peroxidase, and superoxide dismutase. While the HF alone partially reproduced these effects, full antioxidant and cytoprotective activity required the synergistic contribution of both fractions. These findings support the potential application of shrimp waste-derived ethanol extracts as sustainable sources of functional ingredients for oxidative stress-related conditions.

Folate deficiency remains a relevant public health concern, and agronomic biofortification may represent a sustainable strategy to enhance dietary intake through fresh vegetables. This study evaluated the effect of different light intensity regimes on folate accumulation, yield, mineral composition, and nitrate content in mizuna (Brassica rapa L. var. japonica), pakchoi (Brassica rapa L. subsp. chinensis), and chicory (Cichorium intybus L.) grown in a soilless system under LED lighting. Three treatments were applied over a 30day growth cycle: a constant light intensity of 200 µmol m⁻² s⁻¹ (T1); an initial 23day period at 200 µmol m⁻² s⁻¹ followed by a 7day increase to 300 µmol m⁻² s⁻¹ (T2); and a constant light intensity of 300 µmol m⁻² s⁻¹ (T3).Folate content significantly increased under T3 (+24% vs. T1 and T2), particularly in pakchoi, without affecting fresh yield. Higher light intensity also reduced NO3- accumulation (up to −80% in pakchoi), whereas no substantial changes were observed in mineral composition. A 100 g portion of mizuna (T2) or pakchoi (T3) can provide up to 45% of the adult Recommended Daily Allowance (RDA) for folate. Targeted PPFD management therefore represents a promising chemical-free strategy to enhance micronutrient density, although species and genotype selection remain crucial for effective biofortification.

The valorization of agri-food residues is crucial for advancing circular bioeconomy strategies and mitigating environmental impacts. Turnip greens (Brassica rapa subsp. sylvestris) are a traditional vegetable cultivated in southern Italy. While the edible portions include flower sprouts, buds, and young leaves, the more leathery leaves and stems are typically discarded. These wastes represent valuable sources of compounds with antioxidant and antimicrobial potential. This study aims to develop the extraction of phenolic compounds from turnip green residues using two techniques: silent maceration and ultrasound-assisted extraction (UAE). Ethanol was selected over methanol as a food-safe alternative solvent, with preliminary tests confirming equivalent efficiency. A Design of Experiments (DoE) approach was applied to both leaves and stems to assess the effects of solvent composition, solvent-to-matrix ratio, and extraction time on Total Phenolic Content and Trolox Equivalent Antioxidant Capacity. DoE results identified UAE as the most effective method for stems, while for leaves, the solvent-to-dry-mass ratio was the key parameter. HPLC-DAD analysis was performed to identify and quantify the phenolic acids in selected extracts. The antibacterial activity of these extracts against biofilms of six pathogenic strains was evaluated using crystal violet and MTT assays, confirming efficacy in both biofilm formation and mature stages.

Supercritical CO2 (SC-CO2) extraction offers a sustainable method for obtaining solvent-free oils rich in bioactive molecules, valuable for nutraceutical and functional food products. However, the limited stability of certain extract components can hinder their application. This study explores alpha, beta, and gamma cyclodextrins (CDs) as emulsifiers to enhance SC-CO2-extracted tomato oil (TO) stability. TO/CD emulsions with high oil volume fractions (phi) of 60 %, 65 %, 70 %, and 75 % were prepared using the three types of CDs. Only alpha-CDs formed gel-like, stable emulsions up to phi = 70 %. Confocal microscopy revealed that increasing phi led to morphological changes, including reduced droplet size, decreased roundness, and larger coalescence zones, affecting stability and functionality. Emulsions at phi 60 % showed optimal performance with reduced phase separation, high viscosity, smaller droplet size, and lower coalescence. The stability of carotenoids and tocopherols was evaluated under heat (50 degrees C) and UV-C exposure to simulate accelerated aging and sterilization. Emulsions improved carotenoid stability at elevated temperatures compared to bulk TO. Tocopherols were highly stable in bulk TO and moderately stable in TO/alpha-CD emulsions. Under UV-C exposure, TO/alpha-CD emulsions enhanced carotenoid and tocopherol stability for up to 9 h, compared to pure TO. Further analysis with alpha-CDs and synthetic glyceryl trioctanoate (GTO) at phi = 60 %, 65 %, and 70 % replicated the concentrations of lycopene and alpha-tocopherol in TO. These results suggest that TO/alpha-CD emulsions can serve as stable, high-quality ingredients for nutraceutical and functional food applications.

The seafood industry generates large volumes of waste, responsible for serious environmental hazards and high disposal costs. Bioconversion and biorefinery approaches offer effective management of seafood waste, preserving valuable nutrients and supporting sustainable seafood production. Shrimp shells are a rich source of polyunsaturated fatty acids and carotenoids, mainly astaxanthin (AST), whose recovery by means of green strategies represents a challenging goal. In this work, the potential of equimolar binary mixtures of menthol and alkanoic acids (ME:AA) as green solvents for AST recovery from shells of Aristaeomorpha foliacea (Risso, 1827) shrimp has been explored, by evaluating both the yield and the stability of the extracted carotenoid. All tested ME:AA exhibited high efficiency for fast AST extraction in mild conditions, while the mixtures containing acetic (ME:C2) and decanoic (ME:C10) acids ensured the maximum stability of the pigment. HPLC analysis highlighted a variety of extracted AST derivatives, mainly consisting of mono- and diesters of fatty acids with variable side chains. Their overall amount allowed to identify mediterranean A. foliacea as a shrimp species particularly capable of AST uptake and storage. ATR-FTIR measurements pointed out partial deprotonation of the alkanoic acid component following the extraction process, in agreement with the alkalinity of the CaCO3-rich matrix. Interestingly, the antioxidant capacity of the extracts exceeded the predicted one, based on the AST content, suggesting the co-extraction of additional antioxidant compounds. Finally, alpha-cyclodextrin was successfully employed as emulsifying agent to prepare stable ME:C10-in-water microemulsions, which drastically enhanced the chemical stability of extracted AST under standard environmental conditions.

Aiming to address the critical need for renewable energy, this study explores the potential of astaxanthin-rich extracts from shrimp waste as natural sensitizers for dye-sensitized solar cells (DSSCs) using an eco-friendly and economically viable process. Photoconversion efficiencies (PCE) of astaxanthin-based DSSCs were evaluated through I-V measurements while charge transfer resistances (RCT) were assessed by electrochemical impedance spectroscopy. The sensitizer preparation involved extraction of astaxanthin from waste shrimp (Aristaeomorpha foliacea) cephalothoraxes, which were desiccated, finely milled and treated with ethyl acetate, to obtain a crude extract very rich in astaxanthin mainly present in esterified forms. The extraction process was fast and avoided extreme experimental conditions in terms of energy consumption, ultra-vacuum, high temperatures and use of toxic solvents. Direct application of the crude extract as sensitizer for DSSCs resulted in low PCE (0.09 %) and high RCT, due to the limited ability of esterified astaxanthin to interact with the TiO2layer. This limitation was overcome through saponification of the extract with methanolic NaOH, leading to free astaxanthin with high recovery yield and limited degradation. The saponified extract showed lower RCT and higher PCE (0.30 %). These results highlight the promising potential of suitably processed shrimp waste as a source of sensitizers for DSSCs.

Photodynamic therapy (PDT) is a therapeutic option for cancer, in which photosensitizer (PS) drugs, light, and molecular oxygen generate reactive oxygen species (ROS) and induce cell death. First- and second-generation PSs presented with problems that hindered their efficacy, including low solubility. Thus, second-generation PSs loaded into nanocarriers were produced to enhance their cellular uptake and therapeutic efficacy. Among other compounds investigated, the dye methylene blue (MB) showed potential as a PS, and its photodynamic activity in tumor cells was reported even in its nanocarrier-delivered form, including liposomes. Here, we prepared polydopamine (PDA)-coated liposomes and efficiently adsorbed MB onto their surface. lipoPDA@MB vesicles were first physico-chemically characterized and studies on their light stability and on the in vitro release of MB were performed. Photodynamic effects were then assessed on a panel of 2D- and 3D-cultured cancer cell lines, comparing the results with those obtained using free MB. lipoPDA@MB uptake, type of cell death induced, and ability to generate ROS were also investigated. Our results show that lipoPDA@MB possesses higher photodynamic potency compared to MB in both 2D and 3D cell models, probably thanks to its higher uptake, ROS production, and apoptotic cell death induction. Therefore, lipoPDA@MB appears as an efficient drug delivery system for MB-based PDT.

Environmental remediation of heavy metals (HMs) is a crucial aspect of sustainable development, safeguarding natural resources, biodiversity, and the delicate balance of ecosystems, all of which are critical for sustaining life on our planet. The bioremediation of HMs by unicellular phototrophs harnesses their intrinsic detoxification mechanisms, including biosorption, bioaccumulation, and biotransformation. These processes can be remarkably effective in mitigating HMs, particularly at lower contaminant concentrations, surpassing the efficacy of conventional physicochemical methods and offering greater sustainability and cost-effectiveness. Here, we explore the potential of various engineered nanomaterials to further enhance the capacity and efficiency of HM bioremediation based on photosynthetic microorganisms. The critical assessment of the interactions between nanomaterials and unicellular phototrophs emphasised the ability of tailored nanomaterials to sustain photosynthetic metabolism and the defence system of microorganisms, thereby enhancing their growth, biomass accumulation, and overall bioremediation capacity. Key factors that could shape future research efforts toward sustainable nanobioremediation of HM are discussed, and knowledge gaps in the field have been identified. This study sheds light on the potential of nanobioremediation by unicellular phototrophs as an efficient, scalable, and cost-effective solution for HM removal.

Strategies of renewable energy production from photosynthetic microorganisms are gaining great scientific interest as ecosustainable alternatives to fossil fuel depletion. Green microalgae have been thoroughly investigated as living components to convert solar energy into photocurrent in biophotovoltaic (BPV) cells. Conversely, the suitability of diatoms in BPV cells has been almost completely unexplored so far, despite being the most abundant class of photosynthetic microorganisms in phytoplankton and of their good adaptability and resistance to harsh environmental conditions, including dehydration, high salinity, nutrient starvation, temperature, or pH changes. Here, we demonstrate the suitability of a series of diatom species (Phaeodactylum tricornutum, Thalassiosira weissflogii, Fistulifera pelliculosa, and Cylindrotheca closterium), to act as biophotoconverters, coating the surface of indium tin oxide photoanodes in a model BPV cell. Effects of light intensity, cell density, total chlorophyll content, and concentration of the electrochemical mediator on photocurrent generation efficiency were investigated. Noteworthily, biophotoanodes coated with T. weissflogii diatoms are still photoactive after 15 days of dehydration and four rewetting cycles, contrary to analogue electrodes coated with the model green microalga Dunaliella tertiolecta. These results provide the first evidence that diatoms are suitable photosynthetic microorganisms for building highly desiccation-resistant biophotoanodes for durable BPV devices.

All over the world, from America to the Mediterranean Sea, the plant pathogen Xylella fastidiosa represents one of the most difficult challenges with many implications at ecological, agricultural, and economic levels. X. fastidiosa is a rod-shaped Gram-negative bacterium belonging to the family of Xanthomonadaceae. It grows at very low rates and infects a wide range of plants thanks to different vectors. Insects, through their stylets, suck a sap rich in nutrients and inject bacteria into xylem vessels. Since, until now, no antimicrobial treatment has been successfully applied to kill X. fastidiosa and/or prevent its diffusion, in this study, antimicrobial blue light (aBL) was explored as a potential anti-Xylella tool. Xylella fastidiosa subsp. pauca Salento-1, chosen as a model strain, showed a certain degree of sensitivity to light at 410 nm. The killing effect was light dose dependent and bacterial concentration dependent. These preliminary results support the potential of blue light in decontamination of agricultural equipment and/or plant surface; however, further investigations are needed for in vivo applications.

PEGylation is currently the most widespread strategy for the stabilization of nanocarriers for biomedical applications. However, the recognized stealth properties of PEG collide with accelerated blood clearance phenomena and frequently induced side effects. In this work, polydopamine (PDA), a bioinspired polymer, is proposed and evaluated as an alternative to PEG for liposome coating and stabilization. The PDA polymerization conditions are optimized, and the PDA-coated vesicles (Lipo@PDA) are fully characterized from a physicochemical point of view. Protein Corona (PC) formed after vesicle incubation in Fetal Bovine Serum (FBS) is characterized, given that PC determines the behavior of nanocarriers in biological fluids. Proteomic analysis reveals high homology between the most abundant recovered proteins in the PC of Lipo@PDA and PEG-coated liposomes (Lipo@PEG). From a quantitative standpoint, no significant differences were highlighted between the two systems. As for the hemolytic response, it is possible to stay within the 5% red blood cell lysis safety threshold by modulating the concentration and thickness of the PDA shell. Cyto-biocompatibility of Lipo@PDA and Lipo@PEG vesicles towards human respiratory cells NCI–H441 is evaluated. Both kinds of liposomes show a similar behavior: cytotoxicity slightly increases with their concentration. However, biocompatible Lipo@PDA vesicles are obtainable at all tested concentrations by reducing the thickness of the PDA coating. Gathered data suggest that the PDA coating can give the liposomes the same behavior in FBS as the PEG coating, thus offering an opportunity to overcome the drawbacks associated with the use of PEG.

A supramolecular construct for solar energy conversion is developed by covalently bridging the reaction center (RC) from the photosynthetic bacterium Rhodobacter sphaeroides and cytochrome c (Cyt c) proteins with a tailored organic light harvesting antenna (hCy2). The RC-hCy2-Cyt c biohybrid mimics the working mechanism of biological assemblies located in the bacterial cell membrane to convert sunlight into metabolic energy. hCy2 collects visible light and transfers energy to the RC, increasing the rate of photocycle between a RC and Cyt c that are linked in such a way that enhances proximity without preventing protein mobility. The biohybrid obtained with average 1 RC/10 hCy2/1.5 Cyt c molar ratio features an almost doubled photoactivity versus the pristine RC upon illumination at 660 nm, and ∼10 times higher photocurrent versus an equimolar mixture of the unbound proteins. Our results represent an interesting insight into photoenzyme chemical manipulation, opening the way to new eco-sustainable systems for biophotovoltaics.

Attenuated total reflectance Fourier transform infrared (ATR-FTIR) difference spectroscopy has been employed for a variety of applications spanning from reaction mechanisms analysis to interface phenomena assessment. This technique is based on the detection of spectral changes induced by the chemical modification of the original sample. In the present study, we highlight the potential of the ATR-FTIR difference approach in the field of microbial biochemistry and biotechnology, reporting on the identification of main soluble species consumed and released by growing bacteria during the biohydrogen production process. Specifically, the mid-infrared spectrum of a model culture broth, composed of glucose, malt extract and yeast extract, was used as background to acquire the FTIR difference spectrum of the same broth as modified by Enterobacter aerogenes metabolism. The analysis of difference signals revealed that only glucose is degraded during hydrogen evolution in anaerobic conditions, while ethanol and 2,3-butanediol are the main soluble metabolites released with H2. This fast and easy analytical approach can therefore represent a sustainable strategy to screen different bacterial strains and to select raw and waste materials to be employed in the field of biofuel production.

Seed color and size are the major traits influencing consumer's acceptability and market class of lentils worldwide. In this paper we assessed the in vitro antioxidant capacity of whole seeds, hulls, and cotyledons of five lentil varieties in relation to their phenolic profile. The samples were evaluated for total polyphenol content and different phenolic classes, such as condensed tannin content, total monomeric anthocyanins, and phenolic acids. Individual phenolic compounds, including flavonols, flavanols, flavones, anthocyanins, and phenolic acids, were further quantitatively investigated by HPLC-DAD. Total antioxidant capacity was evaluated by ABTS and ORAC assays, and a direct measurement (ABTSdir) was used to evaluate the antioxidant capacity of the bioactive compounds present in the whole-meal flours without extraction. The five genotypes showed considerable variations in their phenolic content and profile as well as antioxidant activities. The results showed a preferential accumulation of phenolic compounds with antioxidant activity in the hulls compared to cotyledons. Delphinidin and cyanidin were the most abundant flavonoids in the hulls, while epicatechin and catechin were the most concentrated in the cotyledons. A highly significant correlation was observed between ABTS, ORAC and ABTSdir and total polyphenols. The antioxidant capacities were highly correlated with several individual phenolics detected in hulls and cotyledons. The overall results showed that the lentil fractions and extracts with higher phenolics had also higher antiradical activity which was independent on seed size and color. Identifying lentil genotypes with diverse phenolic profile in cotyledons and whole seeds could meet diverse consumers preferences and health requirements.

An important research target is improving the health benefits of traditional Mediterranean, durum wheat-based foods using innovative raw materials. In this study, we characterised wholemeal flours obtained from a traditional durum wheat cv. Svevo, two innovative durum wheat varieties (Svevo-High Amylose and Faridur), the naked barley cv. Chifaa and the elite lentil line 6002/ILWL118/1-1, evaluating them for targeted phytochemicals, untargeted metabolomics fingerprints and antioxidant capacity. To this aim, individual phenolic acids, flavonoids, tocochromanols and carotenoids were identified and quantified through HPLC-DAD, and the antioxidant capacities of both the extracts and whole meals were detected by ABTS assays. An untargeted metabolomics fingerprinting of the samples was conducted through NMR spectroscopy. Results showed that the innovative materials improved phytochemical profiles and antioxidant capacity compared to Svevo. In particular, Svevo-HA and Faridur had higher contents of ferulic and sinapic acids, β-tocotrienol and lutein. Moreover, Chifaa is a rich source of phenolic acids, β-tocopherols, lutein and zeaxanthin whereas lentil of flavonoids (i.e., catechin and procyanidin B2). The NMR profiles of Svevo-HA and Faridur showed a significant reduction of sugar content, malate and tryptophan compared to that of Svevo. Finally, substantial differences characterised the lentil profiles, especially for citrate, trigonelline and phenolic resonances of secondary metabolites, such as catechin-like compounds. Overall, these results support the potential of the above innovative materials to renew the health value of traditional Mediterranean durum wheat-based products.