Traditional decoction and PUAE aqueous extracts of pomegranate peels as potential low-cost anti-tyrosinase ingredients

open6The aim of the study is to evaluate the anti-tyrosinase activity of dierent aqueous extracts obtained from pomegranate juice processing by-products. External pomegranate peels of two certified cultivars (Akko and Wonderful), were extracted using only water as the extraction solvent. A traditional decoction and a pulsed ultrasound-assisted extraction (PUAE), both 10 min long, were performed and compared. All the aqueous extracts proved to be rich in bioactive compounds. In particular, the total phenolic content (TPC) ranged from 148 to 237 mg gallic acid equivalent (GAE)/g of dried peels (DW), the radical-scavenging ability (RSA) ranged from 307 to 472 mg ascorbic acid equivalent (AAE)/g DW, the free ellagic acid content (EA) ranged from 49 to 94 g/mL, and the ellagitannins (ETs) ranged from 242 to 340 g/mL. For both cultivars, PUAE extracts had higher ET content and a lower EC50, while the decoctions had slightly higher TPC, RSA, and free EA amounts. Principal component analysis (PCA) highlighted the direct correlation between the ET content and the tyrosinase enzyme inhibition (lower values of EC50). These findings suggest the potential use of both these natural extracts as low-cost lightening and/or anti-browning ingredients exploitable in several formulations (e.g., cosmetics) or extemporarily usable.openFEDERICA TURRINI; PAOLA MALASPINA; PAOLO GIORDANI; SILVIA CATENA; PAOLA ZUNIN; RAFFAELLA BOGGIA;Turrini, Federica; Malaspina, Paola; Giordani, Paolo; Catena, Silvia; Zunin, Paola; Boggia, Raffaell

A Preliminary Color Study of Different Basil-Based Semi-Finished Products during Their Storage

open6: Basil-based semi-finished products, which are mainly used as an intermediate to produce the typical pesto sauce, are prepared and exported all over the world. Color is a fundamental organoleptic requirement for the acceptability of these semi-finished products by the manufacturers of the pesto sauce. Some alternative formulations, which adjust the typical industrial recipe by both changing the preservative agent (ascorbic acid, citric acid, or a mixture of both) and introducing a preliminary thermic treatment (blast chilling), were evaluated. In this work, a fast and non-destructive spectrophotometric analysis, to monitor the color variations in these food products during their shelf-life, was proposed. The raw diffuse reflectance spectra (380–900 nm) obtained by a UV–visible spectrophotometer, endowed with an integrating sphere, together with the CIELab parameters (L*, a*, b*) automatically obtained from these, were considered, and elaborated using multivariate statistical analysis (principal component analysis). From this preliminary study, blast chilling, together with the use of ascorbic acid, proved to be the best solution to better preserve the color of these products during their shelf-life.openTurrini, Federica; Farinini, Emanuele; Leardi, Riccardo; Grasso, Federica; Orlandi, Valentina; Boggia, Raffaella;Turrini, Federica; Farinini, Emanuele; Leardi, Riccardo; Grasso, Federica; Orlandi, Valentina; Boggia, Raffaell

In Silico and In Vitro Study of Antioxidant Potential of Urolithins

In this work, quantum chemical calculations based on density functional theory (DFT) were performed to predict the antioxidant potential of four bioactive gut microbiota metabolites of the natural polyphenols ellagitannins (ETs) and ellagic acid (EA), also known as urolithins (UROs). In order to evaluate their ability to counter the effect of oxidative stress caused by reactive oxygen species (ROS), such as the hydroperoxyl radical (•OOH), different reaction mechanisms were investigated, considering water and lipid-like environments. Through our in silico results, it emerged that at physiological pH, the scavenging activity of all urolithins, except urolithin B, are higher than that of trolox and other potent antioxidants existing in nature, such as EA, α-mangostin, allicin, caffeine and melatonin. These findings were confirmed by experimental assays

Characterization data of water-soluble hydrophilic and amphiphilic dendrimers prodrugs for delivering bioactive chemical entities otherwise non soluble.

More than 40% of bioactive chemical entities (BCEs) developed in pharmaceutical industry are almost water-insoluble, poorly orally bioavailable and/or not via parenteral administrable, and this strongly limits their clinical applications. Drug Delivery (DD) is an engineered technology dealing with the development of delivery systems (DDSs) able to solubilize, transport, target release and maintain therapeutic drugs concentration where needed for long periods. DD frequently makes use of nanosized carriers, often positive charged, including dendrimer such as commercially available and strongly cationic PAMAM and PEI. Nowadays, uncharged dendrimer scaffolds modified with amino acids-modified in their cationic form, are preferred because a more controlled number of nitrogen atoms causes less damage to cells. Then, two hydrophilic (1, 2) [1] (Fig. 1) and three amphiphilic (3-5) [2] (Fig. 2) water-soluble dendrimers were prepared and completely characterized. Once established through proper routine investigations, that these materials could work well as DDSs, they have been used to physically entrap two completely insoluble BCEs i.e. the thiocarbamate (O-TC) 6 [3] and Ellagic Acid (EA) 7 (Fig. 3) with the aim at improving their solubility and in parallel at protecting them from early degradation, at promoting their fast cellular up-take and thus reducing eventual systemic toxicity. Without resorting to toxic excipients and harmful solubilizing agents often used despite the resulting unpleasant side effects, five structurally different nanodispersions (DPXs) loaded with 6 [4] and two with 7 [5] were achieved and completely characterized to confirm their structure and to evaluate their potentiality in biomedical applications

Non-PAMAM amino acids-modified dendrimers nanoparticles for enhancing water-solubility of insoluble bioactive molecules: our state of the art

Non-PAMAM amino acids-modified dendrimers nanoparticles for enhancing water-solubility of insoluble bioactive molecules: our state of the art Silvana Alfei,* Andrea Spallarossa, Silvia Catena, Federica Turrini, Guendalina Zuccari, Anna Pittaluga, Raffaella Boggia Dipartimento di Farmacia, Universit\ue0 di Genova, Viale Cembrano 4, I-16148 Genova, Italy E-mail: [email protected] ABSTRACT Water-solubility is essential for GIT absorbability or parenteral administration of drugs, therefore it is a key parameter to achieve the systemic drug concentration necessary for an effective therapeutic activity. Unfortunately, low aqueous solubility is the major problem with bioactive chemical entities (BCEs), in fact, more than 40% BCEs developed in pharmaceutical industry are practically water-insoluble. As a consequence, great are the research efforts focused on the development of new techniques aiming at enhancing it. Toxic excipients and harmful solubilizing agents were also extensively used for solubilizing and delivering non water-soluble drugs, despite the resulting unpleasant side effects complained of by patients. Nowadays, safer strategies, such as drugs physicochemical modifications or particle size reduction, crystal engineering, salt formation, solid dispersion, use of surfactant and complexation are being exploited. As far as what regards dispersion/complexation techniques, nanoparticles, including dendrimers, are intensely utilized for this purpose, thus in parallel achieving drugs protection from early degradation, more efficient target delivery into cells and tissues and lower systemic toxicity. Synthetic thiocarbamate (O-TC 1) (Fig. 1) is a non-nucleoside HIV-1 reverse transcriptase inhibitor [1] while Ellagic Acid (EA 2) (Fig. 2) is a polyphenol present in some fruits, nuts and seeds endowed with strong antioxidant, anti-inflammatory and other several healthy properties. Unfortunately, both of them are practically insoluble (Table 1), non orally bioavailable, non parenteral administrable, then non usable for therapeutic purposes in their free forms. Fig. 1: Structure of O-TC 1 Fig. 2: Structure of EA 2 Fig. 3: Examples of hydrophilic (left) and amphiphilic (right) dendrimers structure During the last year, these problems have been addressed and successfully resolved by us, and in this communication, the reached promising outcomes have been summarized and the current state of the art provided. Afar from commercially high cytotoxic PAMAM, five non cells-damaging amino acid-modified hydrophilic (3, 4) [2] and amphiphilic (5-7) [3] dendrimers (Fig. 3) have been synthetized and then used as polymer nano-containers to improve 1 and 2 water-solubility. Five (8-12) [4] and two (13, 14) [5] structurally different drugs-loaded nanodispersions (DPXs) were obtained respectively. The structures were confirmed by FT-IR and NMR analysis and all the samples have resulted in being endowed with very good Drug Loading (DL %). Compound 1, totally insoluble except for in highly diluted DMSO when free, once entrapped in dendrimers, shown to be well soluble both in water and in ethanol. In the case of 2, water-solubility was increased even up to 1000 times compared to the free form. For the prerogatives demonstrated in the performed routine analyses, the prepared DPXs could be considered eligible for biomedical and therapeutic applications thus allowing to exploit 1 and 2 pharmacological properties. REFERENCES: 1. A. Spallarossa et al., Eur. J. Med. Chem., 44, 2190 (2009). 2. S. Alfei & S. Catena, Polym. Advan. Technol., 29, 2735 (2018). 3. S. Alfei & S. Catena, Polym. Int., 67, 1572 (2018). 3. S. Alfei et al., Eur. J. Pharm. Sci., 124, 153 (2018). 4. S. Alfei et al., New J. Chem., 2019, DOI: 10.1039/c8nj05657a

Microdispersions of ellagic acid and pomegranate extracts as new potential nutraceutical ingredients

The health properties attributed to several fruits (i.e. pomegranates, raspberries, strawberries, blackberry, chestnuts, walnuts, pecan), herbs (tea) and seeds (berries seeds) are attributed to an important group of natural polyphenols classified as hydrolysable tannins (HT) named Ellagitannins (ETs), that have shown in vitro multi-target biological properties relevant to the treatment of several human diseases. In vivo, ETs are rather not absorbed, and they are hydrolysed providing mainly Ellagic acid (EA). EA is endowed with the same biological properties of ETs and it could be considered as the responsible of their health benefits. Unfortunately, EA cannot be exploited for in vivo applications because of its poor water solubility (9.7 \u3bcg/mL) and accordingly low bioavailability. At first, aiming to increase EA solubility, an EA solid microdispersion (EA-md) was realized by employing only water and low methoxylated pectin, as a food compatible excipient, by applying spray drying technology. EA-md showed a 22% (w/w) Drug Loading (DL), a 30 times improved water solubility maintaining a remarkable radical scavenging activity [1]. It has been analytically characterised and used for in vivo pharmacological treatments in order to evaluate it as potential nutraceutical ingredient. Adult (3-6 months old) and old (20-22 old months) male mice were chronically administered EA-md dissolved in the drinking water (about 150 mg / Kg) for 14 days. During this period, animals were monitored for the spontaneous motor activity and for curiosity before, during and at the end of the EA-md treatment. Adult and old mice were then sacrificed for \u201cex vivo, in vitro\u201d analysis to test the efficiency of noradrenaline release from cortical nerve endings. It is known that noradrenaline exocytosis from cortical nerve endings is significantly impaired during ageing. We found that the chronic administration of EA-md did not alter the noradrenaline exocytosis from cortical nerve endings of adult mice, but significantly recovered the reduced noradrenaline overflow in aged mice. Further investigations are needed to explore the cellular cascade of events accounting for the beneficial effect. In a second step, pomegranate, as a natural source of EA, has been considered to similarly prepare and investigate an analogous formulation. Since pomegranate fruit is recognized as one of the most important sources of ETs, mainly localized in the by-products obtained after industrial juice squeezing, a method to convert the squeezing marcs into a potential nutraceutical ingredient has been explored. In particular, Pulsed Ultrasound-Assisted Extraction (PUAE), using just water as solvent, resulted to be suitable for extracting the water-soluble bioactive molecules (PEx), whose content in hydrolysable tannins, standardized in EA, has been determined. Furthermore, the already mentioned spray drying microdispersion has been employed to formulate and to stabilize it over time. This last formulation (PEx-md) will be subjected to the already mentioned pharmacological experiments in order to study its nutraceutical properties too. [1] S. Alfei, F. Turrini, S. Catena, P. Zunin, B. Parodi, G. Zuccari, A.M. Pittaluga, R. Boggia, New J. Chem, 43, 2438-2448 DOI: 10.1039/C8NJ05657

Cationic Polystyrene-Based Hydrogels as Efficient Adsorbents to Remove Methyl Orange and Fluorescein Dye Pollutants from Industrial Wastewater

Water pollution from dyes is harmful to the environment, plants, animals, and humans and is one of the most widespread problems afflicting people throughout the world. Adsorption is a widely used method to remove contaminants derived from the textile industry, food colorants, printing, and cosmetic manufacturing from water. Here, aiming to develop new low-cost and up scalable adsorbent materials for anionic dye remediation and water decontamination by electrostatic interactions, two cationic resins (R1 and R2) were prepared. In particular, they were obtained by copolymerizing 4-ammonium methyl and ethyl styrene monomers (M1 and M2) with dimethy lacrylamide (DMAA), using N-(2-acryloylamino-ethyl)-acrylamide (AAEA) as cross-linker. Once characterized by several analytical techniques, upon their dispersion in an excess of water, R1 and R2 provided the R1- and R2-based hydrogels (namely R1HG and R2HG) with equilibrium degrees of swelling (EDS) of 900% and 1000% and equilibrium water contents (EWC) of 90 and 91%, respectively. By applying Cross’ rheology equation to the data of R1HG and R2HG’s viscosity vs. shear rate, it was established that both hydrogels are shear thinning fluids with pseudoplastic/Bingham plastic behav ior depending on share rate. The equivalents of -NH3 + groups, essential for the electrostatic-based absorbent activity, were estimated by the method of Gaur and Gupta on R1 and R2 and by potentio metric titrations on R1HG and R2HG. In absorption experiments in bulk, R1HG and R2HG showed high removal efficiency (97–100%) towards methyl orange (MO) azo dye, fluorescein (F), and their mixture (MOF). Using F or MO solutions (pH = 7.5, room temperature), the maximum absorption was 47.8 mg/g in 900 (F) and 47.7 mg/g in 1200 (MO) for R1, while that of R2 was 49.0 mg/g in 200 (F) and 48.5 mg/g in 300 (MO). Additionally, R1HG and R2HG-based columns, mimicking decontamination systems by filtration, were capable of removing MO, F, and MOF from water with a 100% removal efficiency, in different conditions of use. R1HG and R2HG represent low-cost and up-scalable column packing materials that are promising for application in industrial wastewater treatment

Neuroinflammation in Aged Brain: Impact of the Oral Administration of Ellagic Acid Microdispersion

The immune system and the central nervous system message each other to preserving central homeostasis. Both systems undergo changes during aging that determine central age-related defects. Ellagic acid (EA) is a natural product which is beneficial in both peripheral and central diseases, including aging. We analyzed the impact of the oral administration of a new oral ellagic acid micro-dispersion (EAm), that largely increased the EA solubility, in young and old mice. Oral EAm did not modify animal weight and behavioral skills in young and old mice, but significantly recovered changes in "ex-vivo, in vitro" parameters in old animals. Cortical noradrenaline exocytosis decreased in aged mice. EAm administration did not modify noradrenaline overflow in young animals, but recovered it in old mice. Furthermore, GFAP staining was increased in the cortex of aged mice, while IBA-1 and CD45 immunopositivities were unchanged when compared to young ones. EAm treatment significantly reduced CD45 signal in both young and old cortical lysates; it diminished GFAP immunopositivity in young mice, but failed to affect IBA-1 expression in both young and old animals. Finally, EAm treatment significantly reduced IL1beta expression in old mice. These results suggest that EAm is beneficial to aging and represents a nutraceutical ingredient for elders

the 3rs reduction and refinement through a multivariate statistical analysis approach in a behavioural study to unveil anxiolytic effects of natural extracts of tilia tomentosa

We propose a multivariate statistical approach based on Principal Component Analysis (PCA) as an useful instrument to improve the Rules of Refinement and Reduction in in vivo animal experimentation. We analysed with PCA the preliminary data from a study on the effects of the oral administration of Tilia tomentosa bud extracts (TTBEs) on the behavioural skills of adult and aged male and female mice. PCA allows to rationalize the data set information and to dissect the results, showing connections among variables under study (behavioural parameters) and different trends in the experimental groups (control and TTBEs-administered animals). Our results show that PCA can give some important information that can be useful for the refinement of the experimental protocol, in order to reduce the number of the animals used in the experiments and/or the behavioural tests to get reliable information