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Plumeria rubra L. is an ornamental Caribbean plant widely known for its ethnobotanical uses and pharmacological activities. The 'Tonda Palermitana' cultivar, on which no data are to date available, is commonly cultivated in Sicily. The aim of our study was to characterize the micro-morphological features of leaves and flowers of this cultivar by light and Scanning Electron Microscopy and to investigate the phytochemical profile and the biological properties of their food-grade extracts (LE and FE, respectively) by LC-DAD-ESI-MS analysis and different in vitro assays. Numerous branched laticifers were observed, and their secretion contained alkaloids and lipophilic compounds as confirmed by histological analyses. Phytochemical analyses showed the presence of alkaloids (9%), terpenoids (13%) and fatty acids (6%), together with a very abundant presence of iridoids (28%) and polyphenols (39%). The most notable biological activity of both extracts appears to be the antioxidant one, showing half-inhibitory concentrations (IC 50 ) about 5 times lower than those detected in anti-inflammatory assays (383.74 ± 5.65 and 232.05 ± 2.87 vs. 1981.23 ± 12.82 and 1215.13 ± 10.15, for FE and LE, respectively), with LE showing the best, and statistically significant ( p < 0.001), biological activity. These results allow us to speculate promising nutraceutical and cosmeceutical applications for this old Sicilian cultivar.

This study aims to recover the main by-product of Citrus fruits processing, the raw pomace, known also as pastazzo , to produce plant complexes to be used in the treatment of inflammatory bowel disease (IBD). Food-grade extracts from orange (OE) and lemon (LE) pomace were obtained by ultrasound-assisted maceration. After a preliminary phytochemical and biological screening by in vitro assays, primary and secondary metabolites were characterized by proton nuclear magnetic resonance ( 1 H-NMR) and liquid chromatography coupled to diode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI-MS) analyses. The intestinal bioaccessibility and antioxidant and anti-inflammatory properties were investigated by in vitro simulated gastro-intestinal digestion followed by treatments on a lipopolysaccharide (LPS)-stimulated human colorectal adenocarcinoma cell line (Caco-2). The tight junctions-associated structural proteins (ZO-1, Claudin-1, and Occludin), transepithelial electrical resistance (TEER), reactive oxygen species (ROS)-levels, expression of some key antioxidant ( CAT , NRF2 and SOD2 ) and inflammatory ( IL-1β , IL-6 , TNF-α , IL-8 ) genes, and pNFkB p65 nuclear translocation, were evaluated. The OE and LE digesta, which did not show any significant difference in terms of phytochemical profile, showed significant effects in protecting against the LPS-induced intestinal barrier damage, oxidative stress and inflammatory response. In conclusion, both OE and LE emerged as potential candidates for further preclinical studies on in vivo IBD models.

Monarda didyma L. (Lamiaceae) is a medicinal and aromatic herb native to eastern North America and now is also cultivated in Northern Italy, which shows terminal heads of bright scarlet-red flowers, subtended by a whorl of red-tinged leafy bracts. Starting from 2018, M. didyma flowering tops have been included in the Belfrit List of botanicals. However, to date studies on the crude extract of this plant are still lacking. The aim of the present study was to investigate the morphological and anatomical features of the flowering tops and the phytochemical profile of their ethanolic and hydroglyceric extracts (EE and HGE, respectively). HGE was the richest in total phenols (105.75 ± 5.91 vs. 64.22 ± 3.45 mg/100 mL) and especially in flavonoids (71.60 ± 5.09 vs. 47.70 ± 1.27 mg/100 mL), as confirmed also by LC-DAD-ESI-MS. Fifty-three polyphenols were identified and quantified. Even if they showed a common polyphenolic profile, EE and HGE showed quantitative differences. Flavan-3-ols and anthocyanins were the most expressed metabolites in HGE, whereas flavonols were the most expressed metabolites in EE. These features confer to HGE the highest antioxidant, anti-inflammatory, and anti-angiogenic properties, detected by several in vitro and in vivo assays, highlighting a promising use of this plant extract for skincare applications.

Saponaria sicula Raf. grows in Sicily, Sardinia, and Algeria on limestone cliffs and volcanic sands 1300-2500 m above sea level. The aim of the present study was to investigate how the pedo-climatic conditions influence the micromorphological, phytochemical, and biological properties of Sicilian S. sicula leaves collected in the Madonie Mountains (SsM) and on Etna Mt (SsE). Micromorphological investigations revealed that leaves from SsM had a higher amount of calcium oxalate druses in the mesophyll and a more intense blue-green staining with Toluidine blue O, indicating a higher content of polyphenols. These data were confirmed by phytochemical analyses carried out on hydroalcoholic extracts, which showed a higher content of total phenols (8.56 ± 0.57 g GAE/100 g DE) and flavonoids (6.09 ± 0.17 g RE/100 g DE) in SsM. Sixty-four compounds were identified by LC-DAD-ESI-MS analysis with propelargonidin dimer as the most abundant compound (10.49% and 10.19% in SsM and SsE, respectively). The higher polyphenol content of SsM leaves matches also with their biological activity, identifying SsM extract as the strongest plant complex (IC 50 2.75-477.30 µg/mL). In conclusion, the present study experimentally demonstrates that not only climatic differences but also soil characteristics affect the micromorphological, phytochemical, and biological features of this plant species.

The nutritional and health properties of algae make them perfect functional ingredients for nutraceutical and cosmeceutical applications. In this study, the Phaeodactylum tricornutum Bohlin (Phaeodactylaceae), a pleiomorphic diatom commonly found in marine ecosystems, was investigated. The in vitro culture conditions used favoured the fusiform morphotype, characterized by a high accumulation of neutral lipids, as detected by fluorescence microscopy after BODIPY staining. These data were confirmed by HPLC-DAD-APCI-MS/MS analyses carried out on the ethanolic extract (PTE), which showed a high content of xanthophylls (98.99%), and in particular of fucoxanthin (Fx, 6.67 g/100 g PTE). The antioxidant activity (ORAC, FRAP, TEAC and β-carotene bleaching) and photostability of PTE and Fx against UVA and UVB rays were firstly evaluated by in vitro cell-free assays. After this, phototoxicity and photoprotective studies were carried out on in vitro reconstructed human epidermidis models. Results demonstrated that PTE (0.1% Fx) and 0.1% Fx, both photostable, significantly ( p < 0.05) reduce oxidative and inflammatory stress markers (ROS, NO and IL-1α), as well as cytotoxicity and sunburn cells induced by UVA and UVB doses simulating the solar radiation, with an excellent safety profile. However, PTE proved to be more effective than Fx, suggesting its effective and safe use in broad-spectrum sunscreens.

The almond industry produces, by bleaching and stripping, two by-products: blanched skin (BS) and blanch water (BW). The aim of this study was to investigate the nutritional and polyphenolic profile, as well as the antioxidant, antimicrobial, antiviral, and potential prebiotic effects of BS and BW from three different Sicilian cultivars. The total phenols and flavonoids contents were ≥1.72 and ≥0.56 g gallic acid equivalents and ≥0.52 and ≥0.18 g rutin equivalents/100 g dry extract (DE) in BS and BW, respectively. The antioxidant activity, evaluated by 2,2-diphenyl-1-picrylhydrazyl scavenging ability, trolox equivalent antioxidant capacity, ferric-reducing antioxidant power, and oxygen radical absorbance capacity, was ≥3.07 and ≥0.83 g trolox equivalent/100 g DE in BS and BW, respectively. Isorhamnetin-3- O -glucoside was the most abundant flavonoid detected in both by-products. No antimicrobial effect was recorded, whereas BS samples exerted antiviral activity against herpes simplex virus 1 (EC 50 160.96 μg/mL). BS also showed high fibre (≥52.67%) and protein (≥10.99) contents and low fat (≤15.35%) and sugars (≤5.55%), making it nutritionally interesting. The present study proved that the cultivar is not a discriminating factor in determining the chemical and biological properties of BS and BW.

Citrus lumia Risso and Poit. 'Pyriformis' are horticultural varieties of Citrus lumia Risso. The fruit is very fragrant and pear-shaped, with a bitter juice, a floral flavor, and a very thick rind. The flavedo shows enlarged (0.74 × 1.16 mm), spherical and ellipsoidal secretory cavities containing the essential oil (EO), visible using light microscopy, and more evident using scanning electron microscopy. The GC-FID and GC-MS analyses of the EO showed a phytochemical profile characterized by the predominance of D-limonene (93.67%). The EO showed interesting antioxidant and anti-inflammatory activities (IC 50 0.07-2.06 mg/mL), as evaluated by the in vitro cell-free enzymatic and non-enzymatic assays. To evaluate the effect on the neuronal functional activity, the embryonic cortical neuronal networks grown on multi-electrode array chips were exposed to non-cytotoxic concentrations of the EO (5-200 µg/mL). The spontaneous neuronal activity was recorded and the mean firing rate, mean burst rate, percentage of spikes in a burst, mean burst durations and inter-spike intervals within a burst parameter were calculated. The EO induced strong and concentration-dependent neuroinhibitory effects, with IC 50 ranging between 11.4-31.1 µg/mL. Furthermore, it showed an acetylcholinesterase inhibitory activity (IC 50 0.19 mg/mL), which is promising for controlling some of the key symptoms of neurodegenerative diseases such as memory and cognitive concerns.

Current evidence supports the use of extra virgin olive oil (EVOO) and its minor components such as hydroxytyrosol or 3,4-dihydroxyphenyl ethanol (DOPET), to improve cardiovascular and metabolic health. Nevertheless, more intervention studies in humans are needed because some gaps remain in its bioavailability and metabolism. The aim of this study was to investigate the DOPET pharmacokinetics on 20 healthy volunteers by administering a hard enteric-coated capsule containing 7.5 mg of bioactive compound conveyed in EVOO. The treatment was preceded by a washout period with a polyphenol and an alcohol-free diet. Blood and urine samples were collected at baseline and different time points, and free DOPET and metabolites, as well as sulfo- and glucuro-conjugates, were quantified by LC-DAD-ESI-MS/MS analysis. The plasma concentration versus time profiles of free DOPET was analyzed by a non-compartmental approach, and several pharmacokinetic parameters (C max , T max , T 1/2 , AUC 0-440 min , AUC 0-∞ , AUC t-∞, AUC extrap&#95;pred , C last and K el ) were calculated. Results showed that DOPET C max (5.5 ng/mL) was reached after 123 min (T max ), with a T 1/2 of 150.53 min. Comparing the data obtained with the literature, the bioavailability of this bioactive compound is about 2.5 times higher, confirming the hypothesis that the pharmaceutical formulation plays a pivotal role in the bioavailability and pharmacokinetics of hydroxytyrosol.

In this study, Mentha pulegium leaves and flowers harvested in three different Sicilian areas were investigated from a micromorphological, phytochemical and biological point of view. Light and scanning electron microscopy showed the presence of spherocrystalline masses of diosmin both in the leaf epidermal cells and in thin flower petals. Two different chemotypes were identified (I, kaempferide/rosmarinic acid; II, jaceidin isomer A). Phytochemical screening identified plant from collection site II as the richest in total phenolics (16.74 g GAE/100 g DE) and that from collection site I as the richest in flavonoids (46.56 g RE/100 g DE). Seventy-seven metabolites were identified both in flower and leaf extracts. Plant from site II showed the best antioxidant (0.90-83.72 µg/mL) and anti-inflammatory (27.44-196.31 µg/mL) activity expressed as half-maximal inhibitory concentration (IC 50 ) evaluated by DPPH, TEAC, FRAP, ORAC, BSA denaturation and protease inhibition assays. These data were also corroborated by in vitro cell-based assays on lymphocytes and erythrocytes. Moreover, plant of site II showed the best antiangiogenic properties (IC 50 33.43-33.60 µg/mL) in vivo on a chick chorioallantoic membrane. In conclusion, pedoclimatic conditions influence the chemotype and the biological activity of M. pulegium , with chemotype I showing the most promising biological properties., Competing Interests: The authors declare no conflict of interest.

The misidentification between edible and poisonous plants is an increasing problem because of the new trend to collect wild plants, especially by amateur collectors who do not have the botanical skills to distinguish between edible and toxic species. Moreover, morphologically similar species are sometimes responsible for accidental contamination or used in the intentional adulteration of products for human and animal consumption. Laurus nobilis L. (laurel) and Prunus laurocerasus L. (cherry laurel) are typical ornamental shrubs of the Mediterranean region. Laurel is considered a non-toxic plant, widely used as flavorings. Conversely, cherry laurel leaves, morphologically similar to those of laurel, contain toxic cyanogenic glycosides. Considering this, the aim of this study was to carry out an in-depth evaluation of laurel and cherry laurel leaves by using light and scanning electron microscopy coupled with three step phytochemical analyses (qualitative and quantitative colorimetric assays and liquid chromatography). This allowed to highlight the distinguishing features of plant species investigated features such as the venation pattern, presence/absence of nectaries, calcium oxalate crystals, secretory idioblasts, and cyanogenic glycosides. Concluding, this multidisciplinary approach can be useful for the identification of plants but also fragments or pruning residues containing cyanogenic glycosides, in quality control tests, intoxications, and criminal cases.

Alnus glutinosa (namely black alder or European alder) is a tree of the Betulaceae family widely spread through Europe, Southeastern Asia, the Caucasus mountains, and Western Siberia. Its bark is traditionally used for medicinal purposes as an astringent, cathartic, febrifuge, emetic, hemostatic, and tonic, suggesting that it may contain bioactive compounds useful to counteract inflammation. The aim of this study was to investigate the phytochemical profile of A. glutinosa stem bark extract (AGE) by LC-DAD-ESI-MS/MS analysis and to validate some biological activities such as antioxidant, anti-inflammatory and anti-angiogenic properties by in vitro and in vivo models (chick chorioallantoic membranes and zebrafish embryos), that can justify its use against inflammatory-based diseases. The AGE showed a high total phenols content expressed as gallic acid equivalents (0.71 g GAE/g of AGE). Diarylheptanoids have been identified as the predominant compounds (0.65 g/g of AGE) with oregonin, which alone constitutes 74.67% of the AGE. The AGE showed a strong and concentration-dependent antioxidant (IC 50 0.15-12.21 µg/mL) and anti-inflammatory (IC 50 5.47-12.97 µg/mL) activity. Furthermore, it showed promising anti-angiogenic activity, inhibiting both the vessel growth (IC 50 23.39 µg/egg) and the release of an endogenous phosphatase alkaline enzyme (IC 50 44.24 µg/embryo). In conclusion, AGE is a promising source of antioxidant, anti-inflammatory and angio-modulator compounds.

Morning headache (MH) is a frequent condition with a not fully explained pathogenesis. During the past years, several studies have been performed to identify a better diet therapy to reduce the frequency and intensity of headaches. Our study aims to determine if an adequate omega-3 (ω-3) intake in a Modified Mediterranean Diet (MMD) can improve the frequency and intensity of MH and, subsequently, the quality of life. Of the 150 enrolled subjects, 95 met the inclusion criteria (63.3%). Patients were included in the study and randomized into two groups: group A following MMD A and group B following MMD B. The MMD A group was designed to arise at least a 1.5:1 ω-6/ω-3 ratio; the MMD B group was designed to derive at least a 4:1 ω-6/ω-3 ratio. Eighty-four subjects completed the 6 weeks intervention (56%). After 6 weeks (t1), group A showed a significant reduction in both Headache Impact Test-6 and Visual Analog Scale scores ( p < 0.001, p < 0.001, respectively). During dietary treatment, group A progressively dismissed ketoprofen-based pharmacological treatment ( p = 0.002) from baseline to t1. Furthermore, a decrease in the platelet-to-lymphocyte ratio and neutrophil-to-lymphocyte ratio at t1 in group A ( p = 0.02) was observed compared to group B. Concluding, this study provides evidence of a positive impact of ω-3-enriched MMD regimen on the inflammatory status and MH. Clinical trial registration: https://clinicaltrials.gov/ct2/show/NCT01890070, identifier: NCT01890070., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Marchetti, Gualtieri, De Lorenzo, Trombetta, Smeriglio, Ingegneri, Cianci, Frank, Schifano, Bigioni and Di Renzo.)

The SARS-CoV-2 pandemic has caused approximately 6.3 million deaths, mainly due to the acute respiratory distress syndrome or multi-organ failure that characterizes COVID-19 acute disease. Post-acute COVID-19 syndrome, also known as long-COVID, is a condition characterized by a complex of symptoms that affects 10-20% of the individuals who have recovered from the infection. Scientific and clinical evidence demonstrates that long-COVID can develop in both adults and children. It has been hypothesized that multi-organ effects of long-COVID could be associated with the persistence of virus RNA/proteins in host cells, but the real mechanism remains to be elucidated. Therefore, we sought to determine the effects of the exogenous expression of the papain-like protease (PLpro) domain of the non-structural protein (NSP3) of SARS-CoV-2 in polarized human airway (Calu-3), intestinal (Caco-2), and liver (HepG2) epithelial cells, and to evaluate the ability of the natural antioxidant hydroxytyrosol (HXT) in neutralizing these effects. Our results demonstrated that PLpro was able to induce a cascade of inflammatory genes and proteins (mainly associated with the interferon pathway) and increase the apoptotic rate and expression of several oxidative stress markers in all evaluated epithelial cells. Noteably, the treatment with 10 μM HXT reverted PL-pro-dependent effects almost completely. This study provides the first evidence that SARS-CoV-2 PLpro remaining in host cells after viral clearance may contribute to the pathogenetic mechanisms of long-COVID. These effects may be counteracted by natural antioxidants. Further clinical and experimental studies are necessary to confirm this hypothesis.

Peucedanum ostruthium (L.) W. D. J. Koch (Apiaceae) is a worldwide perennial herb native to the mountains of central Southern Europe. The rhizome has a long tradition in popular medicine, while ethnobotanical surveys have revealed local uses of leaves for superficial injuries. To experimentally validate these uses, plant material was collected in the Gran Paradiso National Park, Aosta Valley, Italy, and the rhizome and leaves were micromorphologically and phytochemically characterized. Polyphenol-enriched hydroalcoholic rhizome and leaf extracts, used in cell-free assays, showed strong and concentration-dependent antioxidant and anti-inflammatory activities. In vitro tests revealed cyclooxygenase and lipoxygenase inhibition by the leaf extract, while the rhizome extract induced only lipoxygenase inhibition. MTT assays on HaCaT keratinocytes and L929 fibroblasts showed low cytotoxicity of extracts. In vitro scratch wound test on HaCaT resulted in a strong induction of wound closure with the leaf extract, while the effect of the rhizome extract was lower. The same test on L929 cells showed similar wound closure induction with both extracts. The results confirmed the traditional medicinal uses of the rhizome as an anti-inflammatory and wound healing remedy for superficial injuries but also highlighted that the leaves can be exploited for these purposes with equal or superior effectiveness.

Recently, there has been a growing interest in the recovery of agri-food waste within the circular economy perspective. In this study, the nutritional, phytochemical, and biological features of the cold-pressed hempseed oil (HSO) and hempseed meal (HSM) of two industrial hemp varieties (USO 31 and Futura 75, THC ≤ 0.2%) were evaluated. The HSOs showed a high total phenols and flavonoid content, which were confirmed by LC-DAD-ESI-MS analysis, with rutin as the most abundant compound (56.93-77.89 µg/100 FW). They also proved to be a rich source of tocopherols (81.69-101.45 mg/100 g FW) and of a well-balanced ω-6 to ω-3 fatty acid ratio (3:1) with USO 31, which showed the best phytochemical profile and consequently the best antioxidant activity (about two times higher than Futura 75). The HSMs still retained part of the phytochemicals identified in the HSOs (polyphenols, tocopherols, and the preserved ω-6/ω-3 fatty acids ratio) and a modest antioxidant activity. Furthermore, they showed a very interesting nutritional profile, which was very rich in proteins (29.88-31.44 g/100 g FW), crude fibers (18.39-19.67 g/100 g), and essential and non-essential amino acids. Finally, only a restrained amount of anti-nutritional factors (trypsin inhibitors, phytic acid, and condensed tannins) was found, suggesting a promising re-use of these byproducts in the nutraceutical field.