Metabolomic analysis and bioactivities of Arbutus unedo leaves harvested across the seasons in different natural habitats of Sardinia (Italy).

BACKGROUND: Arbutus unedo L. is a wild tree of Mediterranean regions used as food and in traditional medicine and important for afforestation programs. There is no detailed information available on the variation of A. unedo leaves metabolome across the seasons. The leaves were analyzed by Proton nuclear magnetic resonance (1 H NMR)-based metabolomics, comparing samples harvested across the seasons and in ten different natural habitats of Sardinia (Italy). RESULTS: Multivariate analysis showed the impact of seasonal variation on the metabolome: glucose and quinic acid increased in summer, while in spring sucrose was accumulated. ß-Arbutin, the main known active principle of A. unedo, generally reached the highest concentration in autumn. In winter, O-ß-methylglucose, γ-aminobutyric acid (GABA), flavonols (quercetin-3-O-α-rhamnoside, myricetin-3-O-α-rhamnoside, kaempferol-3-O-α-rhamnoside), catechin, and gallocatechin increased. Characteristic metabolomic features were found also for samples collected in different locations. For instance, trees growing at the highest altitude and exposed to lower temperatures produced less flavonols and catechins. The only sample collected on trees growing on limestones, dolomites, and dolomitic limestones type of soil showed generally the highest content of arbutin. The highest phenolics content was found during spring, while samples collected on flowering branches in winter were the ones with the highest flavonoid content. The antioxidant activity was also variated, ranging from 1.3 to 10.1 mg of Trolox equivalents (TE)/mL of extract, and it was positively correlated to both total phenolics and flavonoid content. Winter samples showed the lowest antibacterial activity, while summer and autumn ones exhibited the highest activity (IC50 values ranging from 17.3 to 42.3 µg/mL against Staphylococcal species). CONCLUSION: This work provides 1 H-NMR fingerprinting of A. unedo leaves, elucidating the main metabolites and their variations during seasons. On the basis of arbutin content, autumn could be considered the balsamic period of this taxon. Samples collected in this season were also the most active ones as antibacterial. Moreover, an interesting metabolomic profile enriched in catechins and flavonols was observed in leaves collected in winter on flowering branches which were endowed with high antioxidant potential.

In vitro α-glucosidase inhibition by Brazilian medicinal plant extracts characterised by ultra-high performance liquid chromatography coupled to mass spectrometry.

Aiming at finding natural sources of antidiabetics agents, 15 extracts from Brazilian medicinal plants of the Atlantic Forest and Amazon region were tested against α-glucosidase enzyme. Plants were selected based on the taxonomic relationships with genera including several species with antidiabetic activity. In this screening, the extracts obtained from the flowers of Hyptis monticola and the leaves of Lantana trifolia and Lippia origanoides resulted endowed with promising anti-α-glucosidase activity. The extracts from H. monticola and from L. origanoides collected in two different areas, were characterised by ultra-high performance liquid chromatography coupled to mass spectrometry. Bioassay-guided fractionation led to the identification of several enzyme inhibiting compounds, among them the mechanism of action of naringenin and pinocembrin was investigated. The two L. origanoides extracts showed differences in bioactivity and in the phytochemical profiles. The fractionation of the extract from H. monticola led to a partial loss of the inhibitory effect.

Extraction, Encapsulation into Lipid Vesicular Systems, and Biological Activity of Rosa canina L. Bioactive Compounds for Dermocosmetic Use.

Valorization of wild plants to obtain botanical ingredients could be a strategy for sustainable production of cosmetics. This study aimed to select the rosehip extract containing the greatest amounts of bioactive compounds and to encapsulate it in vesicular systems capable of protecting their own antioxidant activity. Chemical analysis of Rosa canina L. extracts was performed by LC-DAD-MS/MS and 1H-NMR and vitamins, phenolic compounds, sugars, and organic acids were detected as the main compounds of the extracts. Liposomes, prepared by the film hydration method, together with hyalurosomes and ethosomes, obtained by the ethanol injection method, were characterized in terms of vesicle size, polydispersity index, entrapment efficiency, zeta potential, in vitro release and biocompatibility on WS1 fibroblasts. Among all types of vesicular systems, ethosomes proved to be the most promising nanocarriers showing nanometric size (196 ± 1 nm), narrow polydispersity (0.20 ± 0.02), good entrapment efficiency (92.30 ± 0.02%), and negative zeta potential (-37.36 ± 0.55 mV). Moreover, ethosomes showed good stability over time, a slow release of polyphenols compared with free extract, and they were not cytotoxic. In conclusion, ethosomes could be innovative carriers for the encapsulation of rosehip extract.

Plant Secondary Metabolites: An Opportunity for Circular Economy.

Moving toward a more sustainable development, a pivotal role is played by circular economy and a smarter waste management. Industrial wastes from plants offer a wide spectrum of possibilities for their valorization, still being enriched in high added-value molecules, such as secondary metabolites (SMs). The current review provides an overview of the most common SM classes (chemical structures, classification, biological activities) present in different plant waste/by-products and their potential use in various fields. A bibliographic survey was carried out, taking into account 99 research articles (from 2006 to 2020), summarizing all the information about waste type, its plant source, industrial sector of provenience, contained SMs, reported bioactivities, and proposals for its valorization. This survey highlighted that a great deal of the current publications are focused on the exploitation of plant wastes in human healthcare and food (including cosmetic, pharmaceutical, nutraceutical and food additives). However, as summarized in this review, plant SMs also possess an enormous potential for further uses. Accordingly, an increasing number of investigations on neglected plant matrices and their use in areas such as veterinary science or agriculture are expected, considering also the need to implement "greener" practices in the latter sector.

Screening of ninety herbal products of commercial interest as potential ingredients for phytocosmetics.

Elastase and tyrosinase are important targets both for cosmetics and for dermatological disorders. In this work, ninety herbal products were tested as inhibitors of these two enzymes. Eleven extracts resulted strongly active. Four out of them (Camellia sinensis, Ginkgo biloba, Rhodiola rosea, Vitis vinifera) inhibited both enzymes, five (Glycyrrhiza glabra, Ribes nigrum, Rheum officinale, Salvia officinalis, Tilia platyphyllos) were active against tyrosinase only, and two (Ceterach officinarum and Cinnamomum zeylanicum) proved selectively active against elastase. The IC50 ranged from 3.1 to 104.9 µg/mL and 19.3 to 164.3 µg/mL, against elastase and tyrosinase, respectively. The most active extracts resulted enriched in flavonoids (from 1.47 to 56.47 mg RE/g of extract) and phenolics (from 37.43 to 123.56 mg GAE/g of extract), indicating also an antioxidant potential. Finally, a positive correlation between enzymatic bioactivities and phenolic content was also established.

Identification of a Collagenase-Inhibiting Flavonoid from Alchemilla vulgaris Using NMR-Based Metabolomics.

This paper describes the use of 1H NMR profiling and chemometrics in order to facilitate the selection of medicinal plants as potential sources of collagenase inhibitors. A total of 49 plants with reported ethnobotanical uses, such as the healing of wounds and burns, treatment of skin-related diseases, rheumatism, arthritis, and bone diseases, were initially chosen as potential candidates. The in vitro collagenase inhibitory activity of hydroalcoholic extracts of these plants was tested. Moreover, their phytochemical profiles were analyzed by 1H NMR and combined with the inhibitory activity data by an orthogonal partial least squares model. The results showed a correlation between the bioactivity and the concentration of phenolics, including flavonoids, phenylpropanoids, and tannins, in the extracts. Considering the eventual false-positive effect on the bioactivity given by tannins, a tannin removal procedure was performed on the most active extracts. After this procedure, Alchemilla vulgaris was the most persistently active, proving to owe its activity to compounds other than tannins. Thus, this plant was selected as the most promising and further investigated through bioassay-guided fractionation, which resulted in the isolation of a flavonoid, quercetin-3-O-ß-glucuronide, as confirmed by NMR and HRMS spectra. This compound showed not only a higher activity than other flavonoids with the same aglycone moiety, but was also higher than doxycycline (positive control), the only Federal Drug Administration-approved collagenase inhibitor. The approach employed in this study, namely the integration of metabolomics and bioactivity-guided fractionation, showed great potential as a tool for plant selection and identification of bioactive compounds in natural product research.

Hemidesmus indicus (L.) R. Br. extract inhibits the early step of herpes simplex type 1 and type 2 replication.

Herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) cause several clinically relevant syndromes in both adults and neonates. Despite the availability of efficient anti-HSV agents, the search for new therapeutic approaches is highly encouraged due to the increasing drug resistance of virus strains. Medicinal plants represent a source of potential bioactive compounds. In this context we evaluated the anti-herpetic activity of Hemidesmus indicus (L.) R. Br., a plant widely used in traditional Indian medicine. The hydroalcoholic extract prepared from roots was characterized by NMR and HPLC analysis and assayed in vitro by CPE reduction and virus infectivity assays to define its anti-viral effect. The extract's mechanism of action was investigated by virucidal and time-of-addition assays and by in vitro α-glucosidase inhibitory assay. The extract exhibited a remarkable anti-herpetic activity at 100 mg/mL, at non-cytotoxic concentration, through multiple mechanisms: it reduced the infectivity of viral particles released from infected cells possibly through its anti-ER α-glucosidase inhibitory activity and it inhibited the beginning stage of HSV infection acting as a virucide agent and/or preventing virus attachment to the host cell surface.

Analytical Profiling of Bioactive Phenolic Compounds in Argan (Argania spinosa) Leaves by Combined Microextraction by Packed Sorbent (MEPS) and LC-DAD-MS/MS.

INTRODUCTION: The argan tree (Argania spinosa) is an endemic species from south-western Morocco. Argan-based preparations have been widely used in Moroccan traditional medicine for their biological properties, as well as for several cosmetic purposes. Whereas kernel, pulp of fruit and trunk have been extensively studied for their nutritional and pharmacological effects, relatively little is known about argan tree leaves. OBJECTIVE: The main purpose of the present study is to investigate and characterise the bioactive phenolic fractions in both crude and aqueous extracts derived from argan tree leaves. METHODOLOGY: A qualitative profile of the antioxidant phenolic compounds in argan leaves was obtained by means of structural hypothesis based on UV spectra and mass spectrometric fragmentation patterns. Moreover, selected phenolics were quantified in argan leaves by using a fully validated method based on liquid chromatography coupled to diode array detection and tandem mass spectrometry (LC-DAD-MS/MS). All the extracts were purified by a fast and reliable microextraction by packed sorbent (MEPS) procedure, before analysing them by LC-MS/MS. RESULTS: Based on retention times, mass spectrometric fragmentation and UV spectra, 13 phenolic compounds were identified or tentatively elucidated from crude and aqueous extracts derived from Argania spinosa leaves, while seven compounds were quantified in both extracts. CONCLUSION: The obtained results could represent a first step towards a complete characterisation of the argan plant, its bioactive profiling and the valorisation of its by-products as a source of potentially beneficial bioactive molecules.

Antioxidant and α-glucosidase inhibitory activities of Achillea tenorii.

CONTEXT: There is a need for the discovery of novel natural remedies to prevent and treat metabolic disorders such as hyperglycemia, type II non-insulin-dependent diabetes mellitus, and obesity. Several Achillea species have been utilized for centuries all around the world and are generally considered effective as hypoglycemic. OBJECTIVE: Considering the ethnobotanical uses of Achillea genus, we evaluated the in vitro inhibitory activity of Achillea tenorii Grande (Asteraceae) extract on α-glucosidase, which is a valuable target to prevent and treat metabolic disorders. We also tested its antioxidant activity. Moreover, the phytochemical profile was discussed from a chemotaxonomic point of view. MATERIALS AND METHODS: In vitro α-glucosidase inhibition of crude ethanolic extract obtained from the aerial parts was assayed as well as the in vitro antioxidant activity (ABTS, DPPH, and FRAP-FZ tests) was measured. The extract was characterized from a phytochemical point of view by means of spectroscopic analysis. RESULTS: The extract results endowed with α-glucosidase inhibitory activity (IC50 32 µg/mL) with a particular mechanism of action definable as un-competitive, which differed from the mechanism observed for the best-known α-glucosidase inhibitor (acarbose and miglitol). In addition, a considerable antioxidant potential has been found for A. tenorii extract, which resulted mainly constituted by phenolic compounds such as caffeoylquinic acids and flavonoids. DISCUSSION AND CONCLUSIONS: These results suggest the potential of A. tenorii as a possible natural remedy to prevent and treat metabolic disorders of carbohydrates.

Relevant and selective activity of Pancratium illyricum L. against Candida albicans clinical isolates: a combined effect on yeast growth and virulence.

BACKGROUND: Alkaloids present in plants of the Amaryllidaceae family are secondary metabolites of high biological interest, possessing a wide range of pharmacological activities. In the search for new plant-derived compounds with antimicrobial activities, two alkaloid extracts obtained from bulbs and leaves of Pancratium illyricum L., a plant of the Amarillydaceae family, were tested for their effect on bacterial and yeast growth. METHODS: The broth microdilution susceptibility test was applied to study the effect of plant extracts on the growth of reference bacterial strains and Candida albicans reference and clinical isolates strains. Extracts obtained from the different parts of the plant were tested and compared with the pure components identified in the extracts. Since matrix metalloproteinase enzymes play a role in the dissemination process of Candida albicans, the effect of the bulb extract and pure alkaloids on in vitro collagenase activity was tested. Cell viability test was carried out on human embryo lung fibroblasts (HEL 299). RESULTS: Whilst both extracts did not show any inhibitory activity against neither Gram positive nor Gram negative bacteria, a strong antifungal activity was detected, in particular for the bulb extract. All clinical isolates were susceptible to the growth inhibitory activity of the bulb extract, with endpoint IC50 values ranging from 1.22 to 78 µg/mL. The pure alkaloids lycorine and vittatine, identified as components of the extract, were also assayed for their capacity of inhibiting the yeast growth, and lycorine turned very active, with endpoint IC50 values ranging from 0.89 to 28.5 µg/mL. A potent inhibition of the in vitro collagenase activity was found in the presence of the bulb extract, and this effect was much higher than that exerted by the pure alkaloids. Viability of cell lines tested was not affected by the extract. CONCLUSIONS: Taken together, results suggest that the extract of Pancratium illyricum may act as antifungal agent both directly on the yeast growth and by altering the tissue invasion process.

Expression of pro-inflammatory interleukin-8 is reduced by ayurvedic decoctions.

Eleven decoctions, obtained from indian plants widely used in ayurvedic medicine, have been investigated as a possible source of molecules exhibiting biological activity on the interaction between DNA and NF-kB, a transcription factor involved in the expression of proinflammatory genes. Cystic fibrosis (CF) cell line stimulated by TNF-α has been used as inflammatory cellular model to determinate interleukin-8 (IL-8), one of the most relevant pro-inflammatory mediator in CF regulated by the NF-kB. The chemical characterization of these 11 decoctions by spectrophotometric analysis and NMR fingerprinting highlighted that sugars and polyphenols seemed to be the main compounds. Our results demonstrated that Azadirachta indica, Terminalia bellerica, Terminalia chebula, Hemidesmus indicus, Emblica officinalis and Swertia chirata are the most active decoctions in inhibiting NF-kB/DNA interactions by EMSA assay and in reducing pro-inflammatory IL- 8 expression in CF cells at IC50 concentrations by Real-Time and Bio-plex analyses. Finally, we observed the increase of all inhibitory activities with the rise of total polyphenols, procyanidins and flavonoids, except for the levels of IL-8 mRNA accumulation, that were as high as flavonoid content grown up by the statistical multivariate analyses. In conclusion, these six decoctions might be interesting to explore new anti-inflammatory treatments for diseases, such as CF.

Saving the local tradition: ethnobotanical survey on the use of plants in Bologna district (Italy).

BACKGROUND: Traditional knowledge about plants is unfortunately subjected to a progressive loss, mainly due to globalization and depopulation of the rural areas. This work enhances the ethnobotanical knowledge from Northern Italy, specifically Bologna district, and contributes to preserving Italy's plant-based traditional knowledge and to valorize local resources also in view of an ecological transition. METHODS: The study was conducted between 2010 and 2016 in Bologna district encompassing 22 municipalities, which were grouped into three areas: hill, mountain, and plain. In total, 1172 key informants were interviewed, ranging in age from 50 to 85 years, and having strong links with traditional activities in the area. RESULTS: The final inventory included 374 taxa belonging to 91 families. Among these, 251 were wild native, 40 wild alien, 74 cultivated and 6 were products bought from the market. Hill, mountain, and plain provided information on 278, 213, and 110 taxa, respectively. The most cited families were Asteraceae, Lamiaceae, and Rosaceae. The information was systematized in 12 use categories (UC): medicinal (MED), food, cosmetic, domestic, superstitious-magical-religious (SMR), agropastoral, craft, repellent-insecticide, veterinary, toxic, games, other uses and information. The most relevant UC were in turn divided into subcategories. A descriptive table with all the results was also created. MED was the most relevant UC (310 taxa), and among the 17 MED subcategories, the most significant ones were: gastroenteric (160 taxa), respiratory (133 taxa), and dermatologic (122 taxa). Food was also relevant (197 taxa, and 16 subcategories), and the widest food subcategory was nutraceutical (98 taxa). In cosmetic, the most relevant subcategory was skin treatment (37 taxa). Within SMR, the majority of the plants were cited to heal a disease in a ritual or superstitious way (15 taxa), while for agropastoral, the majority of the taxa (29) were cited as feed. CONCLUSIONS: The data collected has highlighted a significant traditional use of plants in Bologna district. Some plants or uses emerged for the first time from an ethnobotanical study carried out in Italy. The inclusion of a large number of municipalities and informants enabled the collection of a wide spectrum of data, encompassing various uses, anecdotes, and historical curiosities, which are crucial to preserve from being forgotten.

Citrus Pomace as a Source of Plant Complexes to Be Used in the Nutraceutical Field of Intestinal Inflammation.

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 (1H-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-1ß, 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.

Arbutus unedo L. Fractions Exhibit Chemotherapeutic Properties for the Treatment of Gastrointestinal Stromal Tumors.

Novel treatments in gastrointestinal stromal tumors (GISTs) are essential due to imatinib resistance and the modest results obtained with multi-target tyrosine kinase inhibitors. We investigated the possibility that the hydroalcoholic extract from the leaves of Arbutus unedo L. (AUN) could harbor novel chemotherapeutics. The bio-guided fractionation of AUN led to a subfraction, FR2-A, that affected the viability of both imatinib-sensitive and -resistant GIST cells. Cells treated with FR2-A were positive for Annexin V staining, a marker of apoptosis. A rapid PARP-1 downregulation was observed, although without the traditional caspase-dependent cleavage. The fractionation of FR2-A produced nine further active subfractions (FRs), indicating that different molecules contributed to the effect promoted by FR2-A. NMR analysis revealed that pyrogallol-bearing compounds, such as gallic acid, gallic acid hexoside, gallocatechin, myricetin hexoside, and trigalloyl-glucose, are the main components of active FRs. Notably, FRs similarly impaired the viability of GIST cells and peripheral blood mononuclear cells (PBMCs), suggesting a non-specific mechanism of action. Nevertheless, despite the lack of specificity, the established FRs showed promising chemotherapeutic properties to broadly affect the viability of GIST cells, including those that are imatinib-resistant, encouraging further studies to investigate whether pyrogallol-bearing compounds could represent an alternative avenue in GISTs.

Exploring the Anti-Inflammatory and Antioxidant Potential, Metabolite Composition and Inorganic Profile of Cistus monspeliensis L. Aerial Parts and Roots.

This work focuses on Cistus monspeliensis L. aerial parts (AP) and roots (R) extracts, investigating the anti-inflammatory and antioxidant potential of the two organs in comparison. At dosages between 1.56 and 6.25 µg/mL, both extracts showed a protective effect against LPS inflammatory stimulus on a macrophage cell line (RAW 264.7). Interestingly, only R was able to significantly reduce both IL-1ß and IL-6 mRNA gene expression in the presence of LPS. Moreover, the treatment of a neuroblastoma cell line (SH-SY5Y) with AP and R at 6.25 µg/mL increased the cell survival rate by nearly 20% after H2O2 insult. However, only R promoted mitochondria survival, exhibited a significantly higher production of ATP and a higher activity of the enzyme catalase than the control. Both AP and R had similar primary metabolites; in particular, they both contained 1-O-methyl-epi-inositol. Labdane and methoxylated flavonoids were the most characteristic compounds of AP, while R contained mainly catechins, gallic acid, and pyrogallol derivatives. Considering the importance of elemental composition in plants, the inorganic profile of AP and R was also investigated and compared. No potentially toxic elements, such as Pb, were detected in any sample.

Phytochemical Profile and In Vitro Bioactivities of Plant-Based By-Products in View of a Potential Reuse and Valorization.

Wastes and by-products of plant origin are of particular interest to develop a circular economy approach, which attempts to turn them into resources. In this work, thirty-seven neglected plant matrices, including agricultural residues, pest plants, and by-products from the herbal and food industry were extracted and tested for their in vitro anti-tyrosinase, antioxidant, and antibacterial activity against the phytopathogens Pseudomonas syringae pv. syringae ATCC 19310 and Clavibacter michiganensis subsp. nebraskense ATCC 27822. Antioxidant activity ranged from 0.3 to 5 mg of Tr. eq/mL of plant extract, and extract of Castanea sativa pericarp (Csp), Rosa damascena buds (post-distillation) (Rod), and Prunus amygdalus exocarp and mesocarp (Pam) were the most powerful ones. Csp was also capable of inhibiting tyrosinase (IC50 = 16.5 µg/mL), as well as three distillation by-products, namely: Cupressus sempervirens (Css) (IC50 = 95.5 µg/mL), Salvia officinalis (Sco) (IC50 = 87.6 µg/mL), and Helichrysum italicum (Hei) (IC50 = 90.1 µg/mL). Five residues from distillation showed antibacterial activity against C. michiganensis (MICs ranging from 0.125 to 1 mg/mL), namely: Salvia sclarea L. (Sas), Salvia rosmarinus Schleid (Sar), Sco, Hei, and Css. The 1H NMR fingerprinting of the bioactive matrices was acquired, detecting primary and secondary metabolites (rosmarinic acid, shikimic acid, sclareol, and hydroxycinnamic acids).

Metabolomics and genetics of reproductive bud development in Ficus carica var. sativa (edible fig) and in Ficus carica var. caprificus (caprifig): similarities and differences.

In figs, reproductive biology comprises cultivars requiring or not pollination, with female trees (edible fig) and male trees (caprifig) bearing different types of fruits. Metabolomic and genetic studies may clarify bud differentiation mechanisms behind the different fruits. We used a targeted metabolomic analysis and genetic investigation through RNA sequence and candidate gene investigation to perform a deep analysis of buds of two fig cultivars, 'Petrelli' (San Pedro type) and 'Dottato' (Common type), and one caprifig. In this work, proton nuclear magnetic resonance (1H NMR-based metabolomics) has been used to analyze and compare buds of the caprifig and the two fig cultivars collected at different times of the season. Metabolomic data of buds collected on the caprifig, 'Petrelli', and 'Dottato' were treated individually, building three separate orthogonal partial least squared (OPLS) models, using the "y" variable as the sampling time to allow the identification of the correlations among metabolomic profiles of buds. The sampling times revealed different patterns between caprifig and the two edible fig cultivars. A significant amount of glucose and fructose was found in 'Petrelli', differently from 'Dottato', in the buds in June, suggesting that these sugars not only are used by the ripening brebas of 'Petrelli' but also are directed toward the developing buds on the current year shoot for either a main crop (fruit in the current season) or a breba (fruit in the successive season). Genetic characterization through the RNA-seq of buds and comparison with the literature allowed the identification of 473 downregulated genes, with 22 only in profichi, and 391 upregulated genes, with 21 only in mammoni.

By-Product Extracts from Castanea sativa Counteract Hallmarks of Neuroinflammation in a Microglial Model.

Castanea sativa is very common in Italy, and the large amount of waste material generated during chestnut processing has a high environmental impact. Several studies demonstrated that chestnut by-products are a good source of bioactive compounds, mainly endowed with antioxidant properties. This study further investigates the anti-neuroinflammatory effect of chestnut leaf and spiny bur extracts, together with the deepest phytochemical characterisation (by NMR and MS) of active biomolecules contained in leaf extracts, which resulted in being more effective than spiny bur ones. BV-2 microglial cells stimulated with lipopolysaccharide (LPS) were used as a model of neuroinflammation. In BV-2 cells pre-treated with chestnut extracts, LPS signalling is partially blocked via the reduced expression of TLR4 and CD14 as well as the expression of LPS-induced inflammatory markers. Leaf extract fractions revealed the presence of specific flavonoids, such as isorhamnetin glucoside, astragalin, myricitrin, kaempferol 3-rhamnosyl (1-6)(2″-trans-p-coumaroyl)hexoside, tiliroside and unsaturated fatty acids, all of which could be responsible for the observed anti-neuroinflammatory effects. Interestingly, the kaempferol derivative has been identified in chestnut for the first time. In conclusion, the exploitation of chestnut by-products is suitable for the achievement of two goals: satisfaction of consumers' demand for new, natural bio-active compounds and valorisation of by-products.

Metabolomic Study of Dactylis glomerata Growing on Aeolian Archipelago (Italy).

The Aeolian Islands (Italy) are a volcanic archipelago in the Tyrrhenian Sea comprising seven main islands, among which are two active volcanoes. The peculiar geological features and the wide variety of environments and soils have an important impact on native plants, and in particular, the Aeolian populations of Dactylis glomerata (a perennial cool-season bunchgrass) exhibit remarkable phenotypic variability. Considering that environmental drivers also strongly affect the production of plant metabolites, this work aimed at comparing the metabolomic profiles of D. glomerata (leaves) harvested at different altitudes on four islands of the Aeolian archipelago, namely: Lipari, Vulcano, Stromboli and Panarea. Samples were analyzed by 1H NMR profiling, and data were treated by PCA. Samples collected on Stromboli were very different from each other and from the samples collected in the other islands. Through an Orthogonal Partial Least Squares (OPLS) model, using altitude as the y variable, it emerged that the concentration of proline, glycine betaine, sucrose, glucose and chlorogenic acid of D. glomerata growing on Stromboli decreased at increasing altitude. Conversely, increasing altitude was associated with an increment in valine, asparagine, fumaric acid and phenylalanine.

Effects of LED supplemental lighting on the growth and metabolomic profile of Taxus baccata cultivated in a smart greenhouse.

Light emitting diode (LED) lamps are increasingly being studied in cultivation of horticultural, ornamental and medicinal plants as means to increase yield, quality, stress resistance, and bioactive compounds content. Enhancing the production of metabolites for medicinal or pharmaceutical use by regulating LED intensity and spectra is a challenging subject, where promising results have been achieved. Nevertheless, some species have been poorly investigated, despite their interest as a source of medicinally active substances, with particular reference to LED effects at the plant cultivation level. This study evaluates the effects of supplementary top-light LED treatments on Taxus baccata, one of the main sources of taxane precursors. Blue, red and mixed red-and-blue spectra were tested at 100 µM m-2 s-1. Moreover, 50 and 150 µM m-2 s-1 intensities were tested for the mixed spectrum. All treatments were set for 14 hours a day and were tested against natural light as control treatment, in a controlled environment, from 19 August to 9 December 2019, this latter date representing 112 days after treatment (DAT) began. A smart monitoring and control system powered by environmental and proximal sensors was implemented to assure homogeneity of temperature, humidity, and base natural light for all the treatments. It resulted in negligible deviations from expected values and reliable exclusion of confusing factors. Biometric measurements and 1H-NMR based metabolomic analysis were performed to investigate growth and phytochemical profile throughout the trial. One-way ANOVA showed that supplemental LED lighting increased plant height and number of sprouts. Considering the mixed red-and-blue spectrum, plant height increased almost proportionally from control to 100 µM m-2 s-1 (+20% at 112 DAT), with no further increase at higher intensity. The number of sprouts was strongly enhanced by LED treatments only in the early phase (48.9 vs. 7.5 sprouts in the averaged 50, 100 and 150 µM m-2 s-1 vs. the control at 28 DAT), with no differences related to intensity in the very early stage, and more persisting effects (up to 56 DAT) for higher intensities. After the very early growth stages (28 DAT), plant vigor showed a modest although significant increase over time compared to the control, with no differences related to light intensity (0.81 vs. 0.74 of NDVI in the averaged 50, 100 and 150 µM m-2 s-1 vs. the control, across 56, 84 and 112 DAT). The different spectra tested at 100 µM m-2 s-1 showed no significant differences in growth parameters, except for a slight beneficial influence of blue (alone or with red) compared to only red for sprouting. According to the metabolomic analysis, treated plants at 28 DAT were characterized by the highest content of sucrose and aromatic compounds. Signals of a putative taxane were detected in the 1H NMR profiles of plants, which were compared to the spectrum of baccatin III standard. However, the intensity of these spectral signals was not affected by the treatment, while they increased only slightly during time. Light at 150 µM m-2 s-1 induced the strongest variation in the metabolome. Conversely, light composition did not induce significant differences in the metabolome.