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Since the possibility to biotechnologically produce melanin by Streptomycetes using plant biomass has been so far poorly investigated, Posidonia oceanica egagropili, a marine waste accumulating along the Mediterranean Sea coasts, was explored as a renewable source to enhance extracellular melanin production by Streptomyces roseochromogenes ATCC 13400. Therefore, different amounts of egagropili powder were added to a culture medium containing glucose, malt extract, and yeast extract, and their effect on the melanin biosynthesis was evaluated. A 2.5 g·L-1 supplementation in 120-h shake flask growths at 26 °C, at pH 6.0 and 250 rpm, was found to enhance the melanin production up to 3.94 ± 0.12 g·L-1, a value 7.4-fold higher than the control. Moreover, 2-L batches allowed to reach a concentration of 9.20 ± 0.12 g·L-1 in 96 h with a productivity of 0.098 g·L-1·h-1. Further studies also demonstrated that the melanin production enhancement was due to the synergistic effect of both the lignin carbohydrate complex and the holocellulose components of the egagropili. Finally, the pigment was purified from the broth supernatant by acidic precipitation and reversed-phase chromatography, characterized by UV absorbance and one- and two-dimensional NMR, and also tested for its chemical, antioxidant, and photo-protective properties. KEY POINTS: • S. roseochromogenes ATCC 13400 produces extracellular soluble melanin. • Egagropili added to the growth medium enhances melanin production and productivity. • Both the lignin carbohydrate complex and the holocellulose egagropili components influence the melanin biosynthesis.

'Asprinio' is a white dry wine characteristic for its acidity and aromatic flavour, known as emerging DOP wine in Southern Italy. Nevertheless, little information is available on the metabolomic profile of this wine. Thus, in this paper we evaluated the colourimetric parameters

Soil rebuilding represents a major challenge in the recovery of abandoned quarries. In this study, we explored the possibility of using reconstructed Technosols, to achieve soil rebuilding goals at an abandoned quarry site. We first investigated the use of a mixture containing commercial manure and limestone debris (LD) as pedotechnomaterials for an “ad hoc” (re)constructed Technosol (CT), for the recovery of an opencast limestone quarry in one of most concentrated quarry areas in the world. In a field experiment, we tested and monitored different pedotechnosystems (PTSs) made up of constructed Technosol + pasture species + different Mediterranean plant species. Specifically, a control (CT, without any additional treatments) was compared to treatments with organic amendment (CTOA) and conventional fertilizers (CTCF). Data were collected over a 12-month period and included crop performance, plant nutritional state, soil physical-chemical parameters, and metabolites. Analysis of variance compared differences among treatments, while factor analysis (FA) interpreted multiple relationships while explaining observed variability. Results showed that CTOA had better soil physical-chemical properties, greater plant growth, and overall superior agronomic performances compared to all other treatments due to the improved substrate conditions. According to FA, these results appear related to the creation of fertile soil conditions, with most of the investigated metabolites (i) playing a pivotal role in observed outcomes, together with (ii) a clear potential in being considered as a reliable fingerprint for investigating plant responses in constructed PTSs. The proposed pedotechniques in CTOA development show a great potential for the full recovery of abandoned limestone quarries in degraded Mediterranean areas by providing an excellent medium for plant growth, facilitating environmental reclamation.

Sumac (Rhus coriaria L.), a spice obtained by grinding whole sumac berries, is a complex natural product with a plethora of biological activities that can be favorably explored in nutraceutical and pharmaceutical fields. Sicilian sumac is herein chemically investigated by means of a combined NMR/HR MS-based profiling. A hydroalcoholic extract was prepared and its complexity unraveled following fractionation in an alcoholic and an aqueous fraction. The 1H-NMR spectrum of this latter fraction was dominated by the signals of gallic acid and its derivatives, whereas HR MS and HR MS/MS analyses highlighted the diversity in malic acid derivatives. Gallotannins and galloyl flavonol glycosides with quercetin and myricetin as the main aglycones were highly represented in the alcoholic fraction. To improve bioavailability of chemicals in sumac extract, the feasibility of a microencapsulation process by spray drying was investigated. In particular, the addition of maltodextrin, cyclodextrin and gum arabic as covering agents was evaluated. The tests were carried out at different temperatures and concentrations. The addition of maltodextrin was found to be suitable as a carrier for the spray drying of sumac extract, having the highest yields, always > 82%. The highest values of a* (red index) and Chroma were recorded with cyclodextrin and maltodextrin at the lowest concentrations and temperatures of spray drying (120°C). The correlations between the characteristics of the studied microcapsules (yields, moisture, total and surface polyphenols and colour) have been represented by Principal Component Analysis (PCA).

Myrtaceae Juss. is a very large family of flowering plants, that are a valuable source of bioactive compounds. From the phytochemical point of view, apart from ubiquitous tannins, flavonoid derivatives and volatile oils, Myrtaceae plants are very rich in phloroglucinol derivatives, divided in two principal subclasses: oligomeric acylphloroglucinols and phloroglucinol-terpene adducts. Despite this large structural variability, many studies on phloroglucinols from the Myrtaceae have focused on unique bioactive acylphloroglucinol compounds, named myrtucommulones. Nevertheless, many studies also reported other interesting phloroglucinol derivatives. Therefore, in this review, a comprehensive description of the chemical features of acylphloroglucinols and meroterpenoid-acylphloroglucinols from the Myrtaceae family is presented along with their biological activities. Moreover, since myrtucommulones and their derivatives have become attractive targets for organic chemists, thanks to their structural features and biological activities, relevant approaches and synthetic procedures are herewith discussed.

Introduction Mediterranean plants are characterised by a high content of bioactive secondary metabolites that play important roles in plant-plant interactions as plant growth regulators and could be useful for the development of new eco-friendly herbicides. Objective An NMR-based metabolomics approach was reported to seek selective phytotoxic plant extracts and putative plant-derived active molecules. Methods Plant extracts derived from five Mediterranean donor species (Pistacia lentiscus, Bellis sylvestris, Phleum subulatum, Petrohrhagia saxifraga and Melilotus neapolitana) were used to treat the hydroponic cultures of three receiving plants (Triticum durum, Triticum ovatum and Avena fatua). Morphological analyses of the treated receiving plants were carried out. NMR-based metabolomics was applied both to characterise the donor plant extracts and to study the effects of the treatments on the receiving plants. Results This study allowed the identification of Melilotus neapolitana and Bellis sylvestris as phytotoxic plant and good candidates for further studies. Specifically, the NMR-based metabolomics investigation showed that these species affect a specific set of metabolites (such as sugars, amino and organic acids) and therefore metabolic pathways [i.e. tricarboxylic acid (TCA) cycle, amino acid metabolism, etc.] that are crucial for the plant growth and development. Moreover, it was possible to identify the metabolite(s) probably responsible for the phytotoxicity of the active extracts. Conclusion The NMR-based metabolomics approach employed in this study led to the identification of two phytotoxic plant extracts and their putative active principles. These new insights will be of paramount importance in the future to find plant derived molecules endowed with phytotoxic activities.

Wastewater irrigation of crops may be effective to avoid depletion (about 70%) of freshwater resources. However, the use of reclaimed waters containing persistent microcontaminants such as antineoplastic drugs is of high environmental concern. These active compounds may affect human health with potentially severe adverse effects. To better understand the impact on human health following irrigation of crops with reused contaminated waters, we exposed four edible plants, Brassica rapa, Lactuca sativa, Raphanus sativus, and Triticum durum, to two commonly used antitumoral drugs: 5-fluorouracil (5-FU), and Cisplatin (CDDP), using metabolomics as a potential functional genomics tool to combine with genotoxicity experiments. The metabolome of the treated and untreated plants was analysed to detect biochemical alterations associated to the exposure, and the potential genotoxic damage related to human exposure to the treated plants was evaluated using the comet assay in human lymphocytes, which are characterized by high sensitivity to genotoxic substances. The edible species were able to assimilate 5-FU and CDDP during the treatment, affecting the biochemical pathways of these plants with subsequent metabolome modifications. These metabolic alterations differed according to the specific species used for the test. Furthermore, all vegetables treated with two concentrations of the selected drugs (10 and 100 μg/L) caused significant (p < 0.0001) genotoxic damage in the cells of the immune system at a higher level than in the lymphocytes directly exposed to single antineoplastic drugs.

Made available in DSpace on 2022-04-28T19:29:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Exploitation of tuff deposits represents an intensive and destructive human activity that can completely modify natural soils, sometimes leading to their total disappearance. Soil reclamation of quarry areas can usually be done by soil rebuilding, employing complex and expensive techniques. In this research, we aimed to evaluate reconstructed Technosols and associated vegetation, called pedotechnosystem (PTS) and their development and behaviour for tuff quarry reclamation for both environmental rehabilitation and agricultural purposes. Technosols were investigated under three different conditions: i) a control consisting of a constructed Technosol (CT) without additional treatments; ii) CT treated with conventional mineral fertilization (CTCF); and, iii) CT treated with a composted mix of commercial organic amendments (CTOA). Different open-air mesocosms (pots) were investigated and evaluated in terms of: i) development, production, and metabolomic profile of rosemary (Rosmarinus officinalis L.), olive (Olea europaea L.), and grapevine (Vitis Vinifera L. ssp. Sativa) in a mixture with a pasture species: ii) the Technosols physical-chemical responses. Specifically, after 15 months, all investigated PTS were evaluated in terms of: i) pasture species total dry matter (TDM) production, height (H), stem diameter (D) and nitrogen (N), phosphorus (P), and potassium (K) content of dry matter; ii) physical-chemical analyses of all investigated substrates; iii) olive, grape, and rosemary characterization by analysing N, P, and K content in dry leaves as well as their metabolomic profile. Statistical differences among treatments were ascertained through ANOVA, while observed variability was interpreted using a principal factor analysis (PFA). Use of CTOA resulted in statistically (p < 0.05) higher performances in terms of TDM as well as P contents in harvested pasture species. In CTOA, more favourable pH conditions (near neutral) for plant growth were observed; additionally, such PTS showed the best performances in terms of soil organic matter (SOM), total N, and P as well as an improvement for most of the investigated organic fractions and relative humification indexes. Identified metabolites varied greatly by species, underscoring the capability of metabolites in fingerprinting different species and related responses to environmental stress factors. Results from PFA showed that: i) CTOA produced better growing conditions and agronomic performance among all investigated PTS; ii) metabolomic profiles are a reliable fingerprint for investigating plant responses to environmental stress and can help differentiate behaviour that follows. Overall, the research demonstrated that CTOA represents a low-cost easily applicable technique for soil rebuilding in tuff quarry areas. Dipartimento di Scienze e Tecnologie Ambientali Biologiche e Farmaceutiche Università degli Studi della Campania “Luigi Vanvitelli”, Via Vivaldi n° 43 Dipartimento di Architettura e Disegno Industriale Università degli Studi della Campania “Luigi Vanvitelli” Abazia di San Lorenzo, via San Lorenzo Istituto di Ricerca sulle Acque Consiglio Nazionale delle Ricerche (IRSA-CNR), V.le F. de Blasio, 5 Department of Plant Protection Rural Engineering and Soils São Paulo State University, Av. Brazil Sul n◦ 56 Center of Nuclear Energy in Agriculture Universidade de São Paulo, Av. Centenário n◦ 303 USDA-NRCS Ecological Sciences Division Dipartimento di Architettura Design e Urbanistica Università Degli Studi di Sassari, Via Piandanna No 4 Desertification Research Centre Università Degli Studi di Sassari, Viale Italia No 39 Department of Plant Protection Rural Engineering and Soils São Paulo State University, Av. Brazil Sul n◦ 56

In the search of new natural products to be explored as possible anticancer drugs, two plant species, namely Ononis diffusa and Ononis variegata, were screened against colorectal cancer cell lines. The cytotoxic activity of the crude extracts was tested on a panel of colon cancer cell models including cetuximab-sensitive (Caco-2, GEO, SW48), intrinsic (HT-29 and HCT-116), and acquired (GEO-CR, SW48-CR) cetuximab-resistant cell lines. Ononis diffusa showed remarkable cytotoxic activity, especially on the cetuximab-resistant cell lines. The active extract composition was determined by NMR analysis. Given its complexity, a partial purification was then carried out. The fractions obtained were again tested for their biological activity and their metabolite content was determined by 1D and 2D NMR analysis. The study led to the identification of a fraction enriched in oxylipins that showed a 92% growth inhibition of the HT-29 cell line at a concentration of 50 µg/mL.

Cannabidiolic acid (CBDA) is the main phytocannabinoid in fiber and seed-oil hemp (Cannabis sativa L.) plants, but its potential health-related capabilities have been masked for years by a greater scientific interest towards its neutral derivative cannabidiol (CBD). This review aims to collect from the literature and critically discuss all the information about this molecule, starting from its biosynthesis, and focusing on its bioactivity, as an anti-inflammatory, anti-emetic, anti-convulsant, and anti-cancerogenic drug. Furthermore, in the awareness that, despite its multiple bioactive effects, currently poor efforts have been made to achieve its reliable purification, herein, we propose a relatively simple, fast, and inexpensive procedure for its recovery from pollen of industrial hemp cultivars. Spectroscopic and spectrometric techniques allowed us to unequivocally identify pure isolated CBDA and to distinguish it from the constitutional isomer tetrahydrocannabinolic acid (THCA-A).

The need for plants to defend themselves, communicate, and somehow contribute to the social life in their ecosystems has triggered the evolution of an astonishing number of diverse chemicals, some of which involved in plant–plant interactions. In the present study, specific aspects of allelopathy are investigated. A combination of bioassays and metabolomics was used in order to study the chemical interactions occurring between three donor species of Mediterranean area (Arbutus unedo, Medicago minima, Myrtus communis) and a receiving species (Aegilops geniculata). The biochemical changes occurring in the receiving plant upon the treatments with the donor extracts were studied. Oxidative stress and altered water balance were found to be the major changes in the receiving plant. Putative allelochemicals synthesized by the donor plants were also identified and it was shown that their activity was enhanced by co-occurring metabolites. This study provides evidence that metabolite mixtures are to be taken into consideration for allelopathic activity. Furthermore, not only it reports the chemicals responsible for the activity in the specific system, but it also shows that the response of the receiving plant to the treatment with extracts from donor plants is comparable to the response to other stresses.

Metabolomics is the analysis of endogenous and exogenous low molecular mass metabolites within cells, tissues, or biofluids of an organism in response to an external stressor. In this chapter, we highlight the importance of the subdiscipline of environmental metabolomics, which investigates the interactions of organisms with environmental stressors such as biotic and abiotic factors, xenobiotics, temperature shifts, and chemical contaminants. Over the past decade, there has been increasing scientific interest in environmental metabolomics, most likely attributable to the comprehensive nature of nontargeted metabolomics. Hypotheses have therefore been developed on complex environmental stressor effects, especially those with unknown modes of action. The availability of a wide variety of model organisms such as freshwater organisms of the food chain has promoted the potential of metabolomics to detect stress from an extensive range of external factors. Furthermore, these dynamics may shift from individuals to populations, contemplating the traditional fields of the ecophysiology and ecology from instantaneous effects to those over evolutionary timescales. In this chapter, we provide an overview of analytical instrumentation, extraction methods, general experimental design, and the statistical methods generally used in environmental metabolomics. Despite the difficulty in understanding the consequences of environmental exposure due to inter- and intra-individual variability, we believe that environmental metabolomics may enrich our understanding of the responses of organisms to the numerous types of environmental stressors.

Ethnopharmacological relevance Allium hooshidaryae (sect. Pseudoprason) is a wild plant in northwestern Iran. The plant is traditionally used, besides as spice, also for its medicinal properties. Aim of the study Due to the shortcoming evidence in scientific research and the importance of this plant in folk medicine, this study aims to assess the chemical compositions and biological activities, which have no longer reported to date. Materials and methods The bulbs of A. hooshidaryae were collected from West Azerbaijan, Iran. The plant essential oil was obtained by hydrodistillation using Clevenger-type apparatus according to the European pharmacopeia. The plant hydromethanolic extract was obtained using maceration method. The volatile oil compositions of A. hooshidaryae bulbs were evaluated by use of combined gas chromatography/flame ionization detector (GC/FID) and gas chromatography/mass spectrometry (GC/MS) techniques. Furthermore, different biological activities of the yielded essential oil and hydromethanolic extract were in vitro evaluated. The antibacterial and antifungal activities were assessed using disc diffusion assay, tube dilution assay, minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), and minimal fungicidal concentration (MFC). The cytotoxic activities were assayed by reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) toward two human cancerous cell lines (MOLT-4 and MCF-7). Antioxidant activity was investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radical scavenging assay. Results GC/FID and GC/MS analyses allowed detecting 62 components in the A. hooshidaryae essential oil representing the 91.87% of the total oil. The volatile compounds were identified by comparison of the relative retention indices (RRI), mass spectra with those in NIST08/NIH and Wiley (257 and 7 L) libraries and co-elution with authentic samples where available. Surprisingly, the most abundant compound was obtained as menthol (19.0%) followed by carvacrol (10.1%), menthone (6.4%), methyl (methylthiomethyl) disulfide (4.2%), dimethyl disulfide (3.8%), and thymol (3.8%). Contrary to the other Allium species enriched by sulfur compounds, just three compounds accounting for 10.7% of the total oil were obtained as the sulfur-sulfur bond containing components (Dimethyl disulfide, Methyl (methylthio) methyl disulfide, Bis-methylthiomethyl disulfide). The hydromethanolic extract of A. hooshidaryae showed higher anti-radical (IC50 DPPH of 9.81 μg/mL) and cytotoxic (for MOLT-4 and MCF-7, IC50s were 76.3 and 128.6 μg/mL, respectively) activities rather than that of the obtained essential oil (IC50 DPPH of 39.9 μg/mL; IC50 MOLT-4 of 109.2 μg/mL, and IC50 MCF-7 of 297.5 μg/mL). While, the essential oil exhibited the anti-Staphylococcus aurous and anti-Escherichia coli activities approximately the same as Chloramphenicol (positive control). The MIC values were 31.25 and 62.5 μg/mL and the disk inhibition zone values were 23 and 21 mm, respectively. In addition, Candida albicans had moderate sensitivity (MFC of 62.5 μg/mL) for the essential oil. Conclusions The hydromethanolic extract of A. hooshidaryae shows the potency to be used for food protection in addition to further cytotoxic investigations. Associated with antimicrobial abilities of both A. hooshidaryae products, the compatible results was observed with the traditional claim having being not investigated to date. These findings will facilitate the development of A. hooshidaryae for further deep investigations.

In this study, the effects of the allelochemical coumarin through a metabolomic, proteomic and morphophysiological approach in Arabidopsis adult plants (25 days old) were investigated. Metabolomic analysis evidenced an increment of amino acids and a high accumulation of soluble sugars, after 6 days of coumarin treatment. This effect was accompanied by a strong decrease on plant fresh and dry weights, as well as on total protein content. On the contrary, coumarin did not affect leaf number but caused a reduction in leaf area. An alteration of water status was confirmed by a reduction of relative water content and an increase in leaf osmotic potential. Moreover, coumarin impaired plant bio-membranes through an increase of lipid peroxidation and H2O2 content suggesting that coumarin treatment might induce oxidative stress. Coumarin reduced the effective quantum yield of the photosystem II, the energy dissipation in the form of heat, the maximum PSII efficiency, the coefficient of the photochemical quenching and the estimated electron transport rate, while it significantly stimulated the fluorescence emission and the coefficient of the non photochemical quenching. Finally, the proteomic characterization of coumarin-treated plants revealed a down-regulation of the ROS detoxifying proteins, responsible of oxidative damage and consequently of physiological cascade effects. (C) 2017 Elsevier GmbH. All rights reserved.

Three unusual amino-phloroglucinols, named helichrytalicines A-C, along to seventeen known compounds including acetophenones, tremetrone derivatives, low-molecular weight phenols, flavonol glucosides, have been isolated from the medium-polar extract of Helichrysum italicum (Roth) G. Don, a medicinal plant typical of the Mediterranean vegetation. The structures of the compounds have been elucidated based on extensive 2D-NMR spectroscopic analyses, including COSY, TOCSY, HSQC, CIGAR-HMBC, H2BC and HSQC-TOCSY, along with Q-TOF HRMS2 analysis. Stereostructure of the new compounds has been elucidated by Mosher's method and NOESY experiment. Antimicrobial properties against Staphylococcus epidermidis of selected compounds have been evaluated. Three unusual amino-phloroglucinols, named helichrytalicines A-C, along with seventeen known compounds including acetophenones, tremetrone derivatives, low-molecular weight phenols, flavonol glucosides, have been isolated from the medium-polar extract of Helichrysum italicum (Roth) G. Don, a medicinal plant typical of the Mediterranean vegetation.The structures of the compounds have been elucidated based on extensive 2D-NMR spectroscopic analyses, including COSY, TOCSY, HSQC, CIGAR-HMBC, H2BC and HSQC-TOCSY, along with Q-TOF HRMS2 analysis. Stereostructure of the new compounds has been elucidated by Mosher's method and NOESY experiment. Antimicrobial properties against Staphylococcus epidermidis of selected compounds have been evaluated. (C) 2016 Elsevier Ltd. All rights reserved.

Astragalus is a cosmopolitan genus characterised by a large number of species with an exceptional rate of diversification. Many of these species are used since ancient times in the Traditional Chinese Medicine (TCM) to cure a condition of general weakness known as “qi”-deficiency. The main phytochemical constituents of these species have been identified as imidazoline alkaloids, nitro toxins, selenium derivatives, polysaccharides, phenols and saponins. Among these, oleanane saponins and cycloartane glycosides are considered as the main responsible for the bioactivity of the plant crude extracts. However, although a plethora of literature data regarding the pharmacological activities of the Astragalus saponins is available, a clear understanding of their mode of action is still missing. In this review, a brief outline of the taxonomy, distribution and morphology of the Astragalus genus is presented along with a comprehensive description of the chemical features and the biological activities of the the saponins from it derived. Moreover, in an attempt to summarise the current mechanistic knowledge on these metabolites, we elaborated a detailed overview of the key signalling pathways that are known to be affected by Astragalus saponins. Finally, we dedicated a section to Astragaloside IV, the best-characterised cycloartane glycoside from Astragalus, to summarise its potentiality as a pharmacological agent. The ultimate goal of this review is to provide the foundations for future investigations, which would eventually enable the introduction of Astragalus saponins in the clinical setting. Astragalus is a cosmopolitan genus characterised by a large number of species with an exceptional rate of diversification. Many of these species are used since ancient times in the Traditional Chinese Medicine (TCM) to cure a condition of general weakness known as "qi"-deficiency. The main phytochemical constituents of these species have been identified as imidazoline alkaloids, nitro toxins, selenium derivatives, polysaccharides, phenols and saponins. Among these, oleanane saponins and cycloartane glycosides are considered as the main responsible for the bioactivity of the plant crude extracts. However, although a plethora of literature data regarding the pharmacological activities of the Astragalus saponins is available, a clear understanding of their mode of action is still missing. In this review, a brief outline of the taxonomy, distribution and morphology of the Astragalus genus is presented along with a comprehensive description of the chemical features and the biological activities of the the saponins from it derived. Moreover, in an attempt to summarise the current mechanistic knowledge on these metabolites, we elaborated a detailed overview of the key signalling pathways that are known to be affected by Astragalus saponins. Finally, we dedicated a section to Astragaloside IV, the best-characterised cycloartane glycoside from Astragalus, to summarise its potentiality as a pharmacological agent. The ultimate goal of this review is to provide the foundations for future investigations, which would eventually enable the introduction of Astragalus saponins in the clinical setting.

The phytochemical profile, antioxidant activity and α-glucosidase inhibitory activity of Hypericum scruglii Bacchetta, Brullo et Salmeri and Hypericum hircinum L. were analyzed and compared to those of the best-known Hypericum perforatum L. Samples were prepared using three different extraction methods (maceration in 70% ethanol, extraction in MeOH/phosphate buffer and decoction), the influence of which on the metabolic profile and bioactivities are discussed. H. scruglii and H. hircinum extracted in 70% ethanol strongly inhibited α-glucosidase (IC50 7.25 and 14.05 μg/ml, respectively), which is a valuable enzymatic target for treating metabolic disorders, while H. perforatum was found to be less powerful than the other two species in all the performed biological tests. The phytochemical profile was analyzed by NMR, HPLC-DAD and HPLC-FLD, revealing remarkable differences among the species. In particular, H. scruglii, which is a species endemic to Sardinia Island (Italy), was particularly enriched in two phloroglucinol-derivatives identified by means of 2D NMR and LC/MS/MS experiments as 3-geranyl-1-(2′-methylbutanoyl)-phloroglucinol and 3-geranyl-1-(2′-methylpropanoyl)-phloroglucinol.

The phytochemical study of Bellis sylvestris has been carried out. This study led to the isolation of 28 secondary metabolites belonging to different classes. The structures of these compounds have been elucidated on the basis of extensive 2D-NMR spectroscopic analyses, including COSY, TOCSY, HSQC, CIGAR-HMBC, H2BC and HSQC-TOCSY, along with Q-TOF HRMS2 analysis. Three of them, two megastimane derivatives and a glycoside of 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone, were reported for the first time. The compounds isolated from Bellis sylvestris were tested for their antimicrobial activity against some microorganisms associated with urinary tract infections (Proteus mirabilis, Pseudomonas aeruginosa, Staphycoccus aureus and Candida albicans). The bacterial strains showed variable degrees of susceptibility to the compounds. Selected compounds were evaluated for their anti-biofilm properties against Pseudomonas aeruginosa and Candida albicans. Caffeic and rosmarinic acids were the once showing a higher reduction rate of biofilm formation. The phytochemical study of Bellis sylvestris has been carried out. This study led to the isolation of 28 secondary metabolites belonging to different classes. The structures of these compounds have been elucidated on the basis of extensive 2D-NMR spectroscopic analyses, including COSY, TOCSY, HSQC, CIGAR-HMBC, H2BC and HSQC-TOCSY, along with Q-TOF HRMS2 analysis. Three of them, two megastimane derivatives and a glycoside of 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone, were reported for the first time.The compounds isolated from Bellis sylvestris were tested for their antimicrobial activity against some microorganisms associated with urinary tract infections (Proteus mirabilis, Pseudomonas aeruginosa, Streptococcus aureus and Candida albicans). The bacterial strains showed variable degrees of susceptibility to the compounds. Selected compounds were evaluated for their anti-biofilm properties against Pseudomonas aeruginosa and Candida albicans. Caffeic and rosmarinic acids were the once showing a higher reduction rate of biofilm formation. (C) 2016 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.

Bacteria are social organisms able to build complex structures, such as biofilms, that are highly organized surface-associated communities of microorganisms, encased within a self- produced extracellular matrix. Biofilm is commonly associated with many health problems since its formation increases resistance to antibiotics and antimicrobial agents, as in the case of Pseudomonas aeruginosa and Staphylococcus aureus, two human pathogens causing major concern. P. aeruginosa is responsible for severe nosocomial infections, the most frequent of which is ventilator-associated pneumonia, while S. aureus causes several problems, like skin infections, septic arthritis, and endocarditis, to name just a few. Literature data suggest that natural products from plants, bacteria, fungi, and marine organisms have proven to be effective as anti-biofilm agents, inhibiting the formation of the polymer matrix, suppressing cell adhesion and attachment, and decreasing the virulence factors’ production, thereby blocking the quorum sensing network. Here, we focus on plant derived chemicals, and provide an updated literature review on the anti-biofilm properties of terpenes, flavonoids, alkaloids, and phenolic compounds. Moreover, whenever information is available, we also report the mechanisms of action.

Few topics are able to channel the interest of researchers, the public, and industries, like nutraceuticals. The ever-increasing demand of new compounds or new sources of known active compounds, along with the need of a better knowledge about their effectiveness, mode of action, safety, etc., led to a significant effort towards the development of analytical approaches able to answer the many questions related to this topic. Therefore, the application of cutting edges approaches to this area has been observed. Among these approaches, metabolomics is a key player. Herewith, the applications of NMR-based metabolomics to nutraceutical research are discussed: after a brief overview of the analytical workflow, the use of NMR-based metabolomics to the search for new compounds or new sources of known nutraceuticals are reviewed. Then, possible applications for quality control and nutraceutical optimization are suggested. Finally, the use of NMR-based metabolomics to study the impact of nutraceuticals on human metabolism is discussed. © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

The discovery of bioactive compounds from natural sources entails an extremely lengthy process due to the timescale and complexity of traditional methodologies. In our study, we used a rapid NMR based metabolomic approach as tool to identify secondary metabolites with anti-proliferative activity against a panel of human colorectal cancer cell lines with different mutation profiles. For this purpose, fourteen Fabaceae species of Mediterranean vegetation were investigated using a double screening method: 1H NMR profiling enabled the identification of the main compounds present in the mixtures, whilst parallel biological assays allowed the selection of two plant extracts based on their strong anti-proliferative properties. Using high-resolution 2D NMR spectroscopy, putative active constituents were identified in the mixture and isolated by performing a bio-guided fractionation of the selected plant extracts. As a result, we found two active principles: a cycloartane glycoside and protodioscin derivative. Interestingly, these metabolites displayed a preferential anti-proliferative effect on colon cancer cell lines with an intrinsic resistance to anti-EGFR therapies. Our work provides an NMR-based metabolomic approach as a powerful and efficient tool to discover natural products with anticancer activities circumventing time-consuming procedures.

An NMR-basedmetabolomic approach has been applied to analyse seven aromaticMediterranean plant species used in traditional cuisine. Based on the ethnobotanical use of these plants, the approach has been employed in order to study the metabolic changes during different seasons. Primary and secondary metabolites have been detected and quantified. Flavonoids (apigenin, quercetin, and kaempferol derivatives) and phenylpropanoid derivatives (e.g., chlorogenic and rosmarinic acid) are the main identified polyphenols. The richness in these metabolites could explain the biological properties ascribed to these plant species. An NMR-based metabolomic approach has been applied to analyse seven aromatic Mediterranean plant species used in traditional cuisine. Based on the ethnobotanical use of these plants, the approach has been employed in order to study the metabolic changes during different seasons. Primary and secondary metabolites have been detected and quantified. Flavonoids (apigenin, quercetin, and kaempferol derivatives) and phenylpropanoid derivatives (e.g., chlorogenic and rosmarinic acid) are the main identified polyphenols. The richness in these metabolites could explain the biological properties ascribed to these plant species.

Allelopathy is the chemical mediated communication among plants. While on one hand there is growing interest in the field, on the other hand it is still debated as doubts exist at different levels. A number of compounds have been reported for their ability to influence plant growth, but the existence of this phenomenon in the field has rarely been demonstrated. Furthermore, only few studies have reported the uptake and the effects at molecular level of the allelochemicals. Allelopathy has been reported on some plants of Mediterranean vegetation and could contribute to structuring this ecosystem. Sixteen plants of Mediterranean vegetation have been selected and studied by an NMR-based metabolomics approach. The extracts of these donor plants have been characterized in terms of chemical composition and the effects on a selected receiving plant, Aegilops geniculata, have been studied both at the morphological and at the metabolic level. Most of the plant extracts employed in this study were found to have an activity, which could be correlated with the presence of flavonoids and hydroxycinnamate derivatives. These plant extracts affected the receiving plant in different ways, with different rates of growth inhibition at morphological level. The results of metabolomic analysis of treated plants suggested the induction of oxidative stress in all the receiving plants treated with active donor plant extracts, although differences were observed among the responses. Finally, the uptake and transport into receiving plant leaves of different metabolites present in the extracts added to the culture medium were observed. © 2014 Elsevier Ltd. All rights reserved. Allelopathy is the chemical mediated communication among plants. While on one hand there is growing interest in the field, on the other hand it is still debated as doubts exist at different levels. A number of compounds have been reported for their ability to influence plant growth, but the existence of this phenomenon in the field has rarely been demonstrated. Furthermore, only few studies have reported the uptake and the effects at molecular level of the allelochemicals.Allelopathy has been reported on some plants of Mediterranean vegetation and could contribute to structuring this ecosystem. Sixteen plants of Mediterranean vegetation have been selected and studied by an NMR-based metabolomics approach. The extracts of these donor plants have been characterized in terms of chemical composition and the effects on a selected receiving plant, Aegilops geniculata, have been studied both at the morphological and at the metabolic level. Most of the plant extracts employed in this study were found to have an activity, which could be correlated with the presence of flavonoids and hydroxycinnamate derivatives. These plant extracts affected the receiving plant in different ways, with different rates of growth inhibition at morphological level.The results of metabolomic analysis of treated plants suggested the induction of oxidative stress in all the receiving plants treated with active donor plant extracts, although differences were observed among the responses. Finally, the uptake and transport into receiving plant leaves of different metabolites present in the extracts added to the culture medium were observed. (C) 2014 Elsevier Ltd. All rights reserved.

A metabolomic-based approach for the study of allelopathic interactions in the Mediterranean area is proposed using Aegilops geniculata Roth (Poaceae), a Mediterranean herbaceous plant, as test species. Its metabolome has been elucidated by 1D and 2D NMR experiments. Hydroponic plant cultures of A. geniculata were treated with specific compounds of known allelopathic potential: catechol, coumarin, p-coumaric acid, p-hydroxybenzoic acid, ferulic acid and juglone. The metabolic variations due to the presence of allelochemicals have been analyzed and measured. All of the compounds showed the strongest effects at the highest concentration, with coumarin and juglone as the most active compounds, causing an increase of several metabolites. The metabolome changes in test plants confirmed the allelochemicals' reported modes of action. The results demonstrated that the proposed method is a promising tool. It can be applied to plant extracts, making it possible to evidence the metabolites responsible for the activity, as well as their mechanisms of action. © 2013 Elsevier B.V.

Four novel γ-pyrone (=4H-pyran-4-one) metabolites, petrorhagiosides A-D, along with four known analogs, have been isolated from the MeOH extract of Petrorhagia saxifraga, a perennial herbaceous plant typical of Mediterranean vegetation. The structures of the new compounds were established on the basis of extensive spectroscopic analyses including 1D- an 2D-NMR (1H, 1H-DQ-COSY, TOCSY, HSQC, CIGAR-HMBC, and HSQC-TOCSY) experiments. Copyright © 2013 Verlag Helvetica Chimica Acta AG, Zürich.

The polyphenolic composition and antioxidant properties of the methanol leaf extract of Vitis × labruscana cv. 'Isabella' has been performed by means of different spectrophotometric antioxidant methods. Intracellular ROS produc- tion was measured by cell fluorescence after loading with 2',7'-H2DCF-DA. The extract promising efficacy has laid the basis to its phytochemical characterization. Three hydroxycinnamoyl tartaric acids (1-3), isoquercitrin 4, and five flavonol glycuronides (5-9) were isolated from Vitis × labruscana cv. 'Isabella' leaves. Chemical structures were elucidated on the basis of their spectroscopic features. Metabolites 6, 7, and 9, characterized by glucuronic or galacturonic acid methyl es- ter, are described from the genus Vitis for the first time. The metabolites isolated as major constituents were investigated for their DPPH radical scavenging capacity.

Golden germander (Teucrium polium L.) is a Mediterranean shrub of the Labiatae family, used in traditional medicine for its diuretic, antipyretic, diaphoretic, antispasmodic, tonic, anti-inflammatory, antihypertensive, anorexic, analgesic, antibacterial and antidiabetic effects. Like other plants of the Teucrium genus, it was widely popular because of its hypoglycemic and hypolipidemic properties but various cases of T. polium-induced hepatitis have been reported. neo-Clerodane diterpenoids, considered chemotaxonomic markers for the Teucrium genus, are believed to be responsible for the observed hepatotoxicity. The plant also produces flavonoids and phenylethanoid glycosides to which the antioxidant and cytoprotective therapeutic properties of its preparations can be traced back. In order to establish a herbal formula that preserves the plant beneficial properties, T. polium leaf drug has been subjected to a bio-guided fractionation. The different phytocomplexes obtained were analyzed by means of an extensive antioxidant screening and hepatotoxicity evaluation against HepG2, a human hepatoblastoma cell line. The cytotoxicity of the fractions was also evaluated against HeLa and A549 cell lines. In order to identify the substances responsible for the bioactivities, NMR-based metabolic profiling techniques of all the phytocomplexes were performed. Data obtained highlighted the possibility of preparing strong antioxidant extracts, useful as food additives, such as MeOH-2, and MeOH-3, completely devoid of hepatotoxic components. © 2012 Elsevier Ltd. All rights reserved.

Nine different training systems for "Annurca Rossa del Sud" apple fruits, including oblique palmette, free palmette, V-shaped, Tatura trellis, Bibaum (R), modified Bibaum (R), triple leader, slender spindle and Solaxe, were evaluated based on agronomic, qualitative and metabolomic traits. Fruits were analysed at harvest and after the reddening process. The slender spindle training system showed the highest cumulative efficiency yield compared to the others. Furthermore, an increase in the content of bioactive compounds in flesh and fruit peels was observed after the reddening process and was influenced by the different training systems. The metabolic variations in apple peel were measured and analysed. Changes in the metabolome highlight the influence of different training systems on apple quality.This multidisciplinary study expands our knowledge of the influence of training systems on a typical Italian apple cultivar. (C) 2016 Elsevier Ltd. All rights reserved.

Teucrium chamaedrys, one of the most common and investigated species of the genus Teucrium, has been used for centuries in traditional medicine for many purposes. Its phytochemical components comprise, among others, phenylethanoid glycosides (PGs) and neo-clerodane diterpenoids. Several reports have demonstrated a wide range of beneficial biological and pharmacological activities of the phenylethanoid components, while the diterpenes were shown to be strongly hepatotoxic. In this work, in vitro cultures were established from leaf explants of T. chamaedrys. Both solid (callus) and liquid (cell suspension) cultures maintained the capacity to produce PGs, with teucrioside (TS) representing the most abundant one. Cell suspensions had a lower TS content than that found in leaf extracts, but higher than that of calli. An NMR-based metabolomics approach was used to compare the product profile of intact plants vs. cell suspension cultures, and results showed that neo-clerodane diterpenes, present in the intact plant, were not detected in cell cultures. Several elicitors were supplied to cell cultures with the aim of increasing TS production, and elicitation was tested at different growth phases and by exposing cells for different periods. Methyl jasmonate and fungal mycelia from Trichoderma viridae and Fusarium moniliforme were able to significantly increase TS production if supplied at the early-exponential growth phase for 24 h. Based on the proposed link between proline and the phenylpropanoid pathways, proline accumulation in cell cultures was followed throughout a 14-day culture period, showing that it strictly reflected that of TS production. Moreover, exogenously supplied proline, and its analogue hydroxyproline, turned out to be very effective in increasing teucrioside production. © 2012 Elsevier Ltd. All rights reserved.

Over time, a large number of cultivars of Olea europaea, one of the oldest cultivated trees in the world, appeared as a result of events such as outcrossing, mutation, and selective pressure (including grower requirements) on the original olive germplasm. Some cultivars are characteristic of a particular country, whereas others are scattered over an area of cultivation. In the Cilento and Vallo di Diano National Park, numerous olive varieties are grown as the native cultivars 'Pisciottana', 'Rotondella', and 'Salella', the varieties 'Frantoio' and 'Biancolilla', widely cultivated in Southern Italy, and a new variety, known as 'FS17', obtained by self-pollination of 'Frantoio' cultivar. In order to assess the phenolic composition of these cultivars, metabolomic techniques (1D and 2D NMR, and HPLC analyses) were extensively applied. 1H-NMR spectral data were processed and analyzed by Principal Component Analysis (PCA) to point out possible differences in polyphenol composition between the cultivars. The radical scavenging capacity of the investigated cultivars was also evaluated through 2,2-diphenyl-1-picrylydrazyl (DPPH). Folin-Ciocalteu reagent was used to estimate the total phenolic content. The cultivar 'FS17' showed lower total polyphenol content and antioxidant activity than the other analyzed varieties. © 2012 Elsevier Ltd.

Eight flavonoid C-glycosides, including three new analogues, have been isolated from leaf and root methanolic extracts of Petrorhagia velutina, a Mediterranean herbaceous plant. The antiproliferative activity against human hepatoblastoma cancer cell line HepG2 has been analyzed by the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) test. Isoorientin (4) significantly reduces the proliferation of HepG2 cells as determined by the complete conversion of the tetrazolium probe into formazan after 48 h of exposure. © 2010 The American Chemical Society and American Society of Pharmacognosy.

From the aerial parts of Petrorhagia velutina, a new dimeric p-coumaroyl glucose and two monomeric cis p-coumaroyl and feruloyl derivatives have been isolated along with three known related compounds. The structural characterization of these compounds has been elucidated by 1D and 2D NMR techniques. CAD mass spectra have allowed to confirm the spectroscopic structural analysis and to disclose the fragmentation pattern of the dimeric p-coumaroyl derivative. Radical scavenging efficacy of all the isolated metabolites was assessed by measuring their ability to scavenge DPPH radical and ABTS radical cation. The dimeric metabolite and the monomeric trans isomers showed an effective reducing power the oxidant probes. © 2009 Phytochemical Society of Europe.