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Dermal fibroblasts are the main actor in many proteins’ secretion, including collagen, preserving skin function. Free radicals are involved in skin aging and damages involving different cellular components. The imbalance between reactive oxygen species (ROS) amount and natural antioxidant enzymes negatively affects skin homeostasis. Natural compounds have recently emerged as a potential anti-aging tool in tissue regeneration. In the present paper we evaluated the antioxidant activity of white and red wines, considering their probable use, as raw materials, for the formulation of cosmetic products with anti-aging properties. We studied a method that would allow the removal of the alcoholic fraction of wines and determined their composition by LC-MS analysis. We then tested the possible cytotoxic effects of red and white wines on fibroblasts by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay, and their antioxidant activity by the catalase activity test in stressing conditions. Finally, we evaluated their anti-aging potential through the β-galactosidase colorimetric assay. Our results showed that wine extracts exhibit a remarkable antioxidant and anti-aging activity, especially on cells exposed to a marked stressful event. These properties could suggest their possible application as cosmetical products for skin regeneration.

(1) Tomentosin is the most representative sesquiterpene lactone extracted by I. viscosa. Recently, it has gained particular attention in therapeutic oncologic fields due to its anti-tumor properties. (2) In this study, the potential anticancer features of tomentosin were evaluated on human Burkitt’s lymphoma (BL) cell line, treated with increasing tomentosin concentration for cytotoxicity screening. (3) Our data showed that both cell cycle arrest and cell apoptosis induction are responsible of the antiproliferative effects of tomentosin and may end in the inhibition of BL cell viability. Moreover, a microarray gene expression profile was performed to assess differentially expressed genes contributing to tomentosin activity. Seventy-five genes deregulated by tomentosin have been identified. Downregulated genes are enriched in immune-system pathways, and PI3K/AKT and JAK/STAT pathways which favor proliferation and growth processes. Importantly, different deregulated genes identified in tomentosin-treated BL cells are prevalent in molecular pathways known to lead to cellular death, specifically by apoptosis. Tomentosin-treatment in BL cells induces the downregulation of antiapoptotic genes such as BCL2A1 and CDKN1A and upregulation of the proapoptotic PMAIP1 gene. (4) Overall, our results suggest that tomentosin could be taken into consideration as a potential natural product with limited toxicity and relevant anti-tumoral activity in the therapeutic options available to BL patients.

Wound-healing is a dynamic skin reparative process that results in a sequence of events, including inflammation, proliferation, and migration of different cell types as fibroblasts. Fibroblasts play a crucial role in repairing processes, from the late inflammatory phase until the fully final epithelization of the injured tissue. Within this context, identifying tools able to implement cell proliferation and migration could improve tissue regeneration. Recently, plants species from all over the world are coming out as novel tools for therapeutic applications thanks to their phytochemicals, which have antioxidant properties and can promote wound healing. In this paper, we aimed at investigating antioxidant activity of waste extracts from different medicinal plants, endemic of the Mediterranean area, on fibroblast proliferation and wound healing. We determined the amount of total phenols and anti-oxidant activity by ABTS assay. We then evaluated the cytotoxicity of the compounds and the proliferative capabilities of fibroblasts by scratch assay. Our results showed that waste extracts retain antioxidant and regenerative properties, inducing tissue re-establishment after environmental stress exposure. Taken together, our findings suggest that waste material could be used in the future also in combinations to stimulate wound healing processes and antioxidant responses in damaged skin.

The production of saffron spice generates large quantities of plant by-products: over 90% of the plant material collected is discarded, and a consideration fraction of this waste is plant stamens. This work investigated the chemical composition and the antimicrobial activities of the non-polar fraction extracted from four different saffron flower stamens. The chemical composition of ethereal extracts of the saffron stamens was qualitatively assessed by means of gas–chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) analyses. These analyses revealed ethereal extracts to possess a high polyunsaturated fatty acid content. In vitro antibacterial activity of stamen extracts showed no large differences between Gram-positive and Gram-negative bacteria in terms of minimal inhibitory concentration (MIC). In food matrix microbial analysis of the bacterial strains belonging to the main foodborne pathogen species, including Staphylococcus aureus DSM 20231, Escherichia coli DSM 30083, and Listeria monocytogenes DSM 20600, using low-fat UHT milk, revealed a statistically significant reduction in the number of cells (particularly for E. coli and S. aureus with a complete elimination of the population of the two target bacteria following incubation in diethyl ether extracts of saffron stamen (DES) at high concentrations tested, both at 37 °C and 6 °C (for 48 h and 7 days, respectively). A synergic effect was observed when the pathogens were incubated at 6 °C with DES. This work shows these by-products to be excellent sources of bioactive compounds, which could be exploited in high-added-value products, such as food, cosmetics, and drugs.

This study was performed to clarify the strategies of Penicillium digitatum during pathogenesis on citrus, assessing, on albedo plugs, the effects of treatment with sodium bicarbonate (NaHCO3 ), at two different pH values (5 and 8.3), on cell-wall-degrading enzyme activity over a period of 72 h.; Results: Treatment with NaHCO3 , under alkaline pH, delayed the polygalacturonase activity for 72 h, or 48 h in the case of the pectin lyase, compared with the control or the same treatment at pH 5. In contrast, pectin methyl esterase activity rapidly increased after 24 h, in plugs dipped in the same solution. In this case, the activity remained higher than untreated or pH 5-treated plugs up to 72 h.; Conclusion: The rapid increase in pectin methyl esterase activity under alkaline conditions is presumably the strategy of the pathogen to lower the pH, soon after the initiation of infection, in order to restore an optimal environment for the subsequent polygalacturonase and pectin lyase action. In fact, at the same time, a low pH delayed the enzymatic activity of polygalacturonase and pectin lyase, the two enzymes that actually cleave the α-1,4-linkages between the galacturonic acid residues. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

Essential oils from 8 Cuminum cyminum local populations collected in Morocco were investigated for their chemical composition and antimicrobial activity. The chemical composition, investigated by gas chromatographic technique, revealed a common fingerprint in all Moroccan samples: twenty-five compounds were identified with γ-terpinen-7-al being the major component in all samples studied. The antimicrobial activity of cumin essential oils were tested against 10 bacterial strains, belonging to 8 different species, and 6 yeast strains, belonging to 4 species. Lactic acid bacteria showed a good resistance to all essential oil tested while overall the cumin essential oils showed a strong antifungal activity that affected both maximum specific growth rate and lag time

The efficacy of different fungicides, belonging to different chemical groups, was in vitro and in vivo tested in glasshouse trials against Allophoma tropica, the causal agent of a new leaf spot on lettuce. The in vitro trials showed that only a few fungicides were effective in inhibiting the mycelial growth of the four tested strains of A. tropica: the most effective fungicide, with the lowest ED50 and MIC values, was penconazole. Prochloraz, propiconazole, boscalid and fludioxonil showed low ED50 values, but high MIC values. The results from the in vivo trials showed that all the tested fungicides significantly reduced Phoma leaf spot incidence and severity, although differences were observed in the achieved disease pressure in the four trials. Generally, in the presence of an average disease severity in the untreated control, the best disease severity reduction (up to 90%) was provided 9 days after the application of fludioxonil, mancozeb, pyraclostrobin + boscalid, propiconazole and iprodione. Fludioxonil and iprodione showed a persistent effect (up to 90%) still being evident 12 days after the application. Generally, in the presence of a high disease severity in the untreated control, 9 days after the application of fludioxonil, mancozeb, iprodione and pyraclostrobin + boscalid provided a disease severity reduction up to 80%. Moreover, copper provided a disease severity reduction from 84 to 89%, applied as preventative treatment, resulting of special interest, particularly for crops grown under organic farming rules.

The chemical composition, antimicrobial activity, and antioxidant activity of Mentha (M.) requienii, M. pulegium L., and M. aquatica (L.) Huds from Sardinia, Italy, were tested. The chemical composition was determined by Gas Chromatography-Mass Spectrometry (GC-MS) and GC-Flame Ionization Detector (GC-FID) analyses. The antiradical activity was assessed by 2,2-Diphenyl-1-picrylhydrazyl and antioxidant capacity assay (ABTS) assay and minimal inhibition concentration was determined by microdilution broth analysis. The major components found in M. requienii EO were pulegone, menthone, and limonene; similarly, the major components of M. pulegium EO were pulegone and iso-menthone. By contrast, the major components of the essential oil (EO) from M. aquatica are terpinyl acetate and eucalyptol. The antioxidant activity of M. aquatica EO was higher than that of M. requienii and M. pulegium. The EO from M. requienii had good antifungal and antioxidant activities. In the concentration range tested, the mint EOs did not show any antibacterial activity against Staphylococcus aureus 20231 Deutsche SammLung von Mikroorganismen und Zellkulturen (DSMZ), Salmonella enterica subsp. bongori serovar 66:z41:- 13772 DSMZ, and four strains of Listeria monocytogenes. Lactic acid bacteria (LAB) strains were not inhibited by the EOs tested, with the exception of Lactobacillus brevis, which showed slight sensitivity to M. pulegium and M. requenii EOs. Yeast strains were in general sensitive to the different Mentha EOs tested and were species and strain dependent.

This study aims to identify and quantify the glucosinolates from different parts of wild radish R. raphanistrum (leaves, flowers, fruits, roots) using liquid chromatography-tandem mass spectrometry. Glucoraphenin is the predominant compound, accounting for about 87% (w/w) of total glucosinolate content, followed by glucobrassicin, glucoraphasatin and glucoraphanin (153 mg 100 g−1, 149 mg 100 g−1 and 141 mg 100 g−1 FW, respectively) in fruits; followed by glucoraphasatin (3 mg 100 g−1 FW) in flowers and by glucobrassicin, 4-hydroxyglucobrassicin and glucoraphasatin (145 mg 100 g−1, 27 mg 100 g−1 and 24 mg 100 g−1 FW, respectively) in leaves. In roots the major glucosinolate is glucoraphasatin (56 mg 100 g−1 FW) followed by the glucoraphenin and methoxyglucobrassicin (16 mg 100 g−1 and 7 mg 100 g−1 FW, respectively). Principal component analysis allowed the discrimination of fruit samples from other parts of the plant for the majority of glucosinolates and the fruits are highlighted as sources of glucosinolates. The results are interesting given that wild radish is one of the most important weeds of crops in the Mediterranean region and is popular for home vegetable production and for its employment in human nutrition both as a food as well as for medicinal purposes.

Hot subdwarfs are helium burning objects in late stages of their evolution. These stars can develop winds driven by light absorption in the lines of heavier elements. The wind strength depends on chemical composition which can significantly vary from star to star. We aim to understand the influence of metallicity on the strength of the winds of the hot hydrogen-rich subdwarfs HD 49798 and BD+18$^\circ\,$2647. We used UV and optical spectra to derive stellar parameters and abundances. For derived stellar parameters, we predicted wind structure (including mass-loss rates and terminal velocities) with our METUJE code. We derived effective temperature $T_\text{eff}=45\,900\,$K and mass $M=1.46\,M_\odot$ for HD 49798 and $T_\text{eff}=73\,000\,$K and $M=0.38\,M_\odot$ for BD+18$^\circ\,$2647. The abundances can be interpreted as a result of interplay between stellar evolution and diffusion. HD 49798 has a strong wind that does not allow for chemical separation and consequently it shows solar chemical composition modified by hydrogen burning. On the other hand, we did not find any wind in BD+18$^\circ\,$2647 and its abundances are therefore most likely affected by radiative diffusion. Accurate abundances do not lead to a significant modification of wind mass-loss rate for HD 49798, because the increase of the contribution of Fe and Ni to the radiative force is compensated by the decrease of the force due to other elements. The resulting wind mass-loss rate $\dot M=2.1\times10^{-9}\,M_\odot\,\text{yr}^{-1}$ predicts an X-ray light curve during the eclipse which closely agrees with observations. On the other hand, the absence of the wind in BD+18$^\circ\,$2647 for accurate abundances is a result of its peculiar chemical composition. Wind models with accurate abundances provide more reliable wind parameters, but the influence of abundances on the wind parameters is limited in many cases., 11 pages, accepted for publication in Astronomy & Astrophysics

A comparison between High-Performance Thin-Layer Chromatography (HPTLC) analysis and Liquid Chromatography High Resolution Mass Spectrometry (LC&ndash, HRMS), coupled with Principal Component Analysis (PCA) was carried out by performing a combined metabolomics study to discriminate Arbutus unedo (A. unedo) plants. For a rapid digital record of A. unedo extracts (leaves, yellow fruit, and red fruit collected in La Maddalena and Sassari, Sardinia), HPTLC was used. Data were then analysed by PCA with the results of the ability of this technique to discriminate samples. Similarly, extracts were acquired by non-targeted LC&ndash, HRMS followed by unsupervised PCA, and then by LC&ndash, HRMS (MS) to identify secondary metabolites involved in the differentiation of the samples. As a result, we demonstrated that HPTLC may be applied as a simple and reliable untargeted approach to rapidly discriminate extracts based on tissues and/or geographical origins, while LC&ndash, HRMS could be used to identify which metabolites are able to discriminate samples.

The fatty acid (FA), polyphenol content and evaluation of the antioxidant capacity of exhausted Myrtus communis berries (EMB) resulting from the production of myrtle liqueur were assessed. All parts of the exhausted berries exhibited high concentrations of carbohydrates, proteins, lipids and phenolic compounds. The lipid fraction contained a high amount of poly unsaturated fatty acids (PUFA), mainly represented by linoleic acid (&gt, 70%). Of the phenolic acids evaluated by liquid chromatography/mass spectrometry, ellagic acid was the most predominant (&gt, 50%), followed by gallic and quinic acids. Quercetin and quercetin3-O-rhamnoside were the most abundant flavonoids. The seed extracts showed a higher antioxidant potential than the pericarp extracts, the same trend was observed for total phenolic compounds evaluated by spectrophotometric assay. The overall high content of bioactive compounds and the high antioxidant potential of this byproduct sustain its suitability for a number of industrial applications, such as a food ingredient in novel foods, an additive in cosmetic formulations or a component of animal feed formulations.

The bioprospecting of several monofloral Moroccan honeys was carried out. The antiradical activity expressed as mmol Trolox equivalents/kg of honey and evaluated by 2,2-diphenyl-1-picrylhydrazyl assay, ranged from 0.15 for euphorb honey to 1.08 for citrus honey. The antioxidant activity expressed as mmol Fe2+/kg and evaluated by ferric ion reducing antioxidant power assay, ranged from 0.96 for euphorb honey to 4.74 for orange honey. The total phenol content was evaluated by colorimetric assay, while the color attributes were evaluated as transmittance data. Significant Pearson correlation factors were found between total polyphenol amount and antioxidant activity and between color attributes and antioxidant activity. Furthermore the chemical composition of volatile organic compounds was determined. The volatile organic compounds chemical composition of the studied honeys was mainly represented by terpene and benzene derivatives, Maillard reaction products, isoprenoids, and hydrocarbons. The volatiles fingerprint, as well as a targeted high-performance liquid chromatography analysis of the polar components, was used to tentatively confirm the declared botanical origin of the samples studied.

Background Increased serum activity of CK isoenzymes and macroenzymes, and in particular of the brain isoenzyme (CK-BB) has been reported in dogs with central nervous system (CNS) disorders. However, no studies on the possible differences in serum activities of CK iso- or macroenzymes (Macro-CK1 and Macro-CK2) in different neurologic diseases are available. Objective The aim of this study was to describe the electrophoretic distribution of CK iso- and macroenzymes in dogs with CNS disorders in order to assess whether this distribution depends on a specific neurologic disease. Methods This study was done on sera from 45 dogs with neurologic diseases (degenerative, n = 7; idiopathic epilepsy [IE], n = 14; inflammatory, n = 16; space occupying lesions [SOL], n = 8) and from 10 clinically healthy dogs. The separation of serum CK isoenzymes and macroenzymes was performed using an automated electrophoretic method already validated in dogs. Results Compared with healthy dogs, dogs with CNS disorders had significantly higher total CK and CK-BB activities, and a significantly lower Macro-CK2 activity (P < .001). Comparison of pathologic subgroups and healthy dogs revealed significant differences (P < .01) in dogs with IE and inflammatory disorders for total CK activity, in all the subgroups for CK-BB (P < .01), and in dogs with IE and SOL for Macro-CK2 (P < .01). Conclusions The results of this study suggest that CK-BB is released by neurons damaged by inflammatory or degenerative conditions or due to compressive effects of SOL. However, the neurologic diseases cannot be differentiated based on CK-BB or Macro-CK2 activities, unless further studies allow the definition of diagnostic thresholds.

The phytochemical content of Fragaria vesca L. (wild strawberry) grown in Campania Region (South Italy) was investigated. Wild and cultivated fruit were collected from different geographic areas and from bothautochthonous and non-autochthonous germplasm. Extracts were submitted to untargeted liquid chromatography coupled to high resolution mass spectrometry (LC–ESI-Orbitrap–MS) for metabolite profiling. 39 different phenolic compounds have been detected and identified in the methanolic extract of spontaneous and cultivated strawberries from different origins. Targeted quantitative analyses of selected known metabolites were performed using UHPLC interfaced to Q-Trap mass spectrometer in Multiple Reaction Monitoring mode. Principal Component Analysis (PCA) was applied to both untargeted and targeted results. PCA was applied to the untargeted profiling data indicating that fruits from locations with different pedoclimatic conditions can be discerned. Results from quantitative analysis indicate that autochthonous strawberries display significantly higher levels of anthocyanins, especially cyanidin 3-O-glucoside, and other phenolic compounds, like gallic acid and procyanidin B1, compared to the one from non-autochthonous germplasm. The targeted and untargeted metabolite profiling approach used in this work is proposed for the assessment of geographical origin of the berries. © 2016 Elsevier Ltd

The stir bar sorptive extraction (SBSE) techniques, coupled with gas chromatography–mass spectrometry, were successfully applied to the study of Eucalyptus honey’s for the determination of volatile organic compounds (VOCs). An optimization of the extraction method was carried out and the variables, NaCl concentration (used as matrix modifier), and the concentration of honey solution were studied targeting the whole VOCs composition. After the evaluation of the experiments, the best condition for the extraction of honey volatile components was 2 mol/L of NaCl and the more concentrated honey solution (0.5 g of honey per mL of water). Additionally, the results were compared with those obtained by two headspace (HS) techniques, namely solid-phase microextraction (SPME) and dynamic headspace (DHS). SBSE volatiles differ qualitatively and quantitatively from those obtained by the SPME and DHS methods. In any event, the chemical composition of Eucalyptus honey volatiles extracted by all three techniques shows the presence of some typical floral markers. Our results confirm a general trend reported in the literature, which show the higher sensitivity of SBSE in the extraction of less volatile compounds in comparison with HS methods.

Epidemics of basil downy mildew (DM) incited by Peronospora belbahrii have been very severe in Italy since 2013, in part due to the very favorable weather conditions, and losses have occurred in many commercial farms, even after repeated mefenoxam treatments. DM populations from basil plants and seeds, which are associated with failure in downy mildew control under field and greenhouse conditions, have been tested for their sensitivity to mefenoxam. Basil plants were inoculated with a sporangial suspension of seven DM populations and treated, before and after inoculation with the pathogen, with different dosages of mefenoxam: 100 μg/ml, which corresponds to the currently applied field dosage, 200 μg/ml and 1000 μg/ml. Azoxystrobin was used at the field dosage as the chemical control. Three out of four DM populations from seeds and two out of three from basil plants were found to be able to infect basil plants in the presence of 100 μg/ml and 200 μg/ml of mefenoxam, while the field dosage of azoxystrobin (186 μg/ml) was found to be completely effective. The sensitive populations of P. belbahrii were completely controlled by the field dosage of both chemicals also 14 days after the last treatment. This study provides new information on the potential risk of introducing mefenoxam-resistant P. belbahrii inoculum in the field through seeds infected by resistant strains.

Capparis spinosa L. (Capparidaceae), also known as caper, is widely known for its very aromatic flower buds (capers),that are largely employed as a flavouring in cooking. Capparis species are regarded as a potential source of important bioactive compounds, in fact, due to their botanical relationship with Brassica species; they contain glucosinolates, secondary plant metabolites, that have been studied for their potential anticarcinogenic properties. In addition, the presence of other numerous beneficial compounds such as polyphenols, alkaloids, lipids, vitamins and minerals have been reported. The aim of this study was to individuate and determinate the principal bioactive compounds occurring in different part (leaves, buds and flowers) of wild and cultivated C. spinosa collected from different area of Sardinia (Italy). Ultra-high performance liquid chromatography-triple quadrupole/linear ion trap tandem mass spectrometry methods were used for identification and simultaneous determination of 27 bioactive molecules. Analysis of different samples revealed qualitative and quantitative differences in the content of flavonoids, glucosinolates, anthocyanins and phenolic acids. In particular, glucocapparin resulted the most abundant with values ranging from 112 to 364 mg/100 g Fresh Weight (FW); followed by rutin with highest value of 126 mg/100 g FW, 4-hydroxyglucobrassicin with highest value of 42 mg/100 g FW and isorhamnetin 3-O-rutinoside with highest value of 24 mg/100 g FW. Based on this metabolomic targeted approach, quantitative results were treated by principal component analysis to explore and visualise correlation and discrimination among collections of C. spinosa samples. Copyright © 2016 John Wiley & Sons, Ltd.

Introduction Myrtus communis L. (Myrtaceae) is a self‐seeded shrub, widespread in Sardinia, with anti‐inflammatory, antiseptic, antimicrobial, hypoglycemic and balsamic properties. Its berries, employed for the production of sweet myrtle liqueur, are characterised by a high content of bioactive polyphenols, mainly anthocyanins. Anthocyanin composition is quite specific for vegetables/fruits and can be used as a fingerprint to determine the authenticity, geographical origin and quality of raw materials, products and extracts. Objective To rapidly analyse and determine anthocyanins in 17 samples of Myrtus communis berries by developing a platform based on the integration of UHPLC–MS/MS quantitative data and multivariate analysis with the aim of extracting the most information possible from the data. Methodology UHPLC‐ESI‐MS/MS methods, working in positive ion mode, were performed for the detection and determination of target compounds in multiple reaction monitoring (MRM) mode. Optimal chromatographic conditions were achieved using an XSelect HSS T3 column and a gradient elution with 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Principal component analysis (PCA) was applied to the quantitative data to correlate and discriminate 17 geographical collections of Myrtus communis. Results The developed quantitative method was reliable, sensitive and specific and was successfully applied to the quantification of 17 anthocyanins. Peonidin‐3‐O‐glucoside was the most abundant compound in all the extracts investigated. Conclusion The developed methodology allows the identification of quali‐quantitative differences among M. communis samples and thus defines the quality and value of this raw material for marketed products. Moreover, the reported data have an immediate commercial value due to the current interest in developing antioxidant nutraceuticals from Mediterranean plants, including Sardinian Myrtus communis.

Inflammatory bowel diseases (IBDs) are chronic disorders characterized by disruption and ulceration of the colonic mucosa or of any part of the digestive tract (Crohn's disease). Antioxidant/anti-inflammatory herbal extract supplementation could represent an innovative approach to contrast IBDs. Clinical trials demonstrated the efficacy of natural formulas, containing chamomile, in patients with gastrointestinal disorders. This is consistent, albeit in part, with the antioxidant and anti-inflammatory properties of chamomile. The aim of the present study was to explore the possible protective role of a chamomile extract, on human colorectal adenocarcinoma HT29 cell, and rat colon specimens treated with lipopolysaccharide (LPS) to induce an inflammatory stimulus, a well established model of acute ulcerative colitis. In this context, the activities of different biomarkers of inflammation and lipid peroxidation such as ROS, myeloperoxidase (MPO), serotonin (5-HT), prostaglandin (PG)E2 , 8-iso-prostaglandin (8-iso-PG)F2α , NF-kB, tumor necrosis factor (TNF)α and interleukin (IL)-6 were assessed. We found that chamomile extract was as effective as sulfasalazine (2 mg/ml) in reducing the production of MPO, 5-HT, IL-6, NF-kB, TNFα, PGE2 and 8-iso-PGF2α , after inflammatory stimulus. The observed modulatory effects support a rationale use of chamomile supplementation as a promising pharmacological tool for the prevention and management of ulcerative colitis in humans. Copyright © 2016 John Wiley & Sons, Ltd.

Essential oils (EOs) from several individuals of Myrtus communis L. (M. communis) growing in different habitats in Sardinia have been studied. The analyses were focused on four groups of samples, namely cultivated and wild M. communis var. melanocarpa DC, characterized by red/purple berries, and cultivated and wild M. communis var. leucocarpa DC, characterized by white berries. Qualitative and quantitative analyses demonstrated different EO fingerprints among the studied samples: cultivated and wild leucocarpa variety differs mainly from the melanocarpa variety by a high amount of myrtenyl acetate (>200 mg/mL and 0.4 mg/mL in leucocarpa and melanocarpa varieties respectively). Conversely, the wild group is characterized by a higher amount, compared with the cultivated species, of linalool (about 110 mg/mL and 20 mg/mL respectively), linalyl acetate (about 24 mg/mL and about 6 mg/mL respectively) whereas EOs of the cultivated plants were rich in pinocarveol-cis compared with wild plants (about 2 mg/mL and about 0.5 mg/mL respectively). Principal component analysis applied to the chromatographic data confirm a differentiation and classification of EOs from the four groups of M. communis plants. Finally, antioxidant activity of the studied EOs shows differences between the various categories of samples.

Contamination by Listeria monocytogenes is a particularly challenging problem in the food industry due to the ability of the bacterium to develop under conditions normally used for food preservation. Here, we show that the gaseous phase of Citrus limon var pompia leaf essential oil (hereafter PLEO) exerts specific anti-Listeria activity on ricotta salata cheese stored at 5 °C. The synergic effect of gaseous PLEO treatment and refrigeration was first confirmed in vitro on L. monocytogenes strains treated for 3 h with gaseous PLEO and then stored at 5 °C. Ricotta cheese was then inoculated with L. monocytogenes strains and subjected to hurdle technology with different concentrations of gaseous PLEO. Cell counts revealed gaseous PLEO to exert a bactericidal effect on L. monocytogenes 20600 DSMZ and a bacteriostatic effect on a mix of L. monocytogenes strains. Scanning and transmission electron microscopy analyses of L. monocytogenes cells suggested that gaseous PLEO targets the bacterial cell wall and plasma membrane. Chemical analyses of the liquid and vapor phases of PLEO indicated linalyl acetate to be the predominant compound, followed by limonene and the two isomers of citral, whereas EO composition analysis, although generally in line with previous findings, showed the presence of linalyl acetate for the first time. Solid-phase microextraction coupled with gas chromatography confirmed the presence of all crude oil components in the headspace of the box.

Helichrysum italicum (Asteraceae) is a small shrub of the Mediterranean area which colonizes a wide range of habitats. In Sardinia, Italy, the species is widely dispersed in different environments ranging from sandy beaches to holm hoak forests 1250 m above sea level (a. s. l.). Recently, the species has been attributed an increasing economical value mainly for its pharmacological, aromatic proprieties and ornamental uses. In this study, the chemical profiles of 146 H. italicum ssp. microphyllum genotypes from two contrasting habitats (seaside and mountains) in Sardinia were explored. Significant differences in volatile fractions between the two groups were found, displaying a correlation between secondary metabolite production and habitat type. Multivariate analysis of variance revealed that monoterpenes, sesquiterpenes (oxygenated/not oxygenated), alcohols, ester were quantitatively differentially produced in the plants growing under different environmental conditions (P Considering the importance of climatic conditions in plant development and growth, the relationship between meteorological variables and chemical compounds of the volatile fraction was explored by principal component analysis. Nerolidol is mostly positively correlated to mean winter temperature, while italicene, bergamotene, nerol and curcumene are positively influenced by spring and summer precipitation. Finally, Mantel test revealed a significant correlation (Spearman, P = 0.002) between chemical and meteorological matrices of Euclidean distances, suggesting that the most diverse chemical profiles belonged to plants from the farther climatic locations.

The origin and taxonomy of Citrus monstruosa (Rutaceae) are currently unknown and thus a nontargeted metabolomic analysis on the flavedo volatiles was performed. Headspace solid‐phase micro extraction coupled with gas chromatography analysis was applied to eight Citrus spp. including Citrus monstruosa, and the chemical composition of the volatile organic compounds (VOC) was analyzed to give a chemical comparison between different Citrus species. Forty‐four metabolites were detected across the species: several monoterpenes were found in all Citrus spp. and among them limonene comes first in all Citrus spp. ranging from 96.7 to 66.7%. Conversely, some molecules were found to be specific for only a few of the samples. The classical univariate evaluation of the VOC data is unlikely to give objective data for the cultivar classification; a multivariate approach, principal component analysis, is an easy to use, powerful tool. Therefore, a chemometric comparison between chromatographic data of all Citrus spp. was performed with the aim of understanding the role of VOC in the discrimination of the eight studied cultivars.

Four experimental trials were carried out in order to evaluate the efficacy of different products (chemicals, resistance inducers and natural products) as well as heat air when applied to basil seeds naturally infested by Peronospora belbahrii, the causal agent of downy mildew of basil, a pathogen which is seed transmitted. Seed quality, as vigour index, was also evaluated. In general, seed treatment had a positive effect on the reduction of disease incidence and on plant biomass at the end of each trial, with very satisfactory results when disease incidence was lower than 10% and still satisfactory results with a higher disease incidence. Although many of the fungicides, resistance inducers and thyme oil treatments tested showed a significant disease reduction compared with the untreated control, the protection offered was only partial. Moreover, the effectiveness of the tested seed treatments varied considerably between the trials. The seeds treatment with heat air (65°C for 10 min) significantly reduced the number of infected plants per container. The interaction effect between the fungicides (at the lowest dosage tested, with the exception of acibenzolar-S-methyl, and dimethomorph), thyme oil and the heat air treatment was also significant. In terms of biomass, most treatments not significantly improved the fresh weight in comparison with the untreated control. The highest biomass was observed in the plots where seeds were dressed with mancozeb combined with heat air. Interestingly, thiram is also effective against other pathogens affecting basil and the thermal treatment and potassium phosphite are fully compatible with the rules of organic farming. Seed dressing represents the starting point of a full integrated approach for downy mildew management.

Saffron may be spoiled by a variety of microorganisms during cultivation, harvesting, and post harvesting. As saffron can be dried and stored in different ways, this preliminary study explored the natural microbiota present in Moroccan saffron when subjected to different drying techniques. An analysis of the carotenoid-derived volatiles present in the saffron was also carried out. The culturable microbiota of the saffron samples dried using different methods, namely in the shade (also called natural), in the sun, or in the oven, were studied using classical and molecular approaches. The effect of the drying methods on head-space chemical volatiles was also determined. Eighty-two isolates grown in the different culture media were chosen from the colonies, and genotype analysis grouped the microorganisms into 58 clusters, revealing a wide diversity. Out of the 82 isolates, 75 belonged to the Bacillaceae family. The other isolates were distributed within the Dietziaceae, Paenibacillaceae and Carnobacteriaceae families. The dominant species was Bacillus simplex, which was detected in all samples, regardless of the drying method used. Lysinibacillus macroides was dominant in the sun-dried saffron. No pathogens were isolated, but an isolate belonging to Dietzia maris, a potential human pathogenic species, was detected. The biodiversity indexes were linked to the drying method and generally decreased as the intensity of the treatment increased. The results of this preliminary work show that the different drying methods strongly influenced the microbiota and affect the saffron volatile profile. Further analysis will be needed to determine possible effects of selected microbiota on saffron volatiles.

The efficacy of the removal of contaminants from wastewater depends on physico-chemical properties of pollutants and the efficiency of treatment plant. Sometimes, low amounts of toxic compounds can be still present in the treated sewage. In this work we considered the effects of contaminant residues in treated wastewaters and of temperatures on Lemna minor L. Treated effluent waters were collected, analyzed and used as duckweed growth medium. In order to better understand the effects of micropollutants and seasonal variation, the plants were grown under ambient conditions for seven days in summer and winter. Relative growth rate, pigments and phenolic compounds concentrations were determined, as well as the activities of catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (G-POD) and polyphenol oxidase (PPO). The pollutant concentrations varied in the two seasons, depending on the industrial and municipal activities and efficiency of treatments. Treated waters contained heavy metals, nitrogenous and phosphorus compounds, surfactants and hydrocarbons. Compared to the control, duckweed growth of treated plants decreased by 25% in summer, while in the winter due to the lower temperatures and the presence of pollutants was completely impeded. The amounts of photosynthetic pigments of treated plants were not significantly affected in the summer, while they were higher than the control in the winter when the effluent had a high nitrogen amount. High CAT activity was registered in both seasons. Treated plants had significantly lower APX activity in the summer (53%) and winter (59%) respect to the controls. The observed inhibition of the peroxidase activities in the exposed plants, confirms the controversy existing in the literature about the variability of enzymatic response in stress condition.

Moringa oleifera is a medicinal plant and an excellent dietary source of micronutrients (vitamins and minerals) and health-promoting phytochemicals (phenolic compounds, glucosinolates and isothiocyanates). Glucosinolates and isothiocyanates are known to possess anti-carcinogenic and antioxidant effects and have attracted great interest from both toxicological and pharmacological points of view, as they are able to induce phase 2 detoxification enzymes and to inhibit phase 1 activation enzymes. Phenolic compounds possess antioxidant properties and may exert a preventative effect in regards to the development of chronic degenerative diseases. The aim of this work was to assess the profile and the level of bioactive compounds in all parts of M. oleifera seedlings, by using different MS approaches. First, flow injection electrospray ionization mass spectrometry (FI-ESI-MS) fingerprinting techniques and chemometrics (PCA) were used to achieve the characterization of the different plant's organs in terms of profile of phenolic compounds and glucosinolates. Second, LC-MS and LC-MS/MS qualitative and quantitative methods were used for the identification and/or determination of phenolics and glucosinolates in M. oleifera.

In Transmissible Spongiform Encephalopathies (TSEs), the localization of the prion protein in the neuronal membrane lipid rafts (LR) seems to play a role in sustaining the protein misfolding. Changes in membrane properties, due to altered lipid composition, affect their organization and interaction between lipids and protein therein, and consequently also membrane resident protein functionality; dietary polyunsaturated fatty acids (PUFAs), gangliosides and cholesterol seem to influence these processes. Aims In this work, the influence of administration of different feed, able to change the composition of lipid membrane, on the clinical progression of prion disease was studied. Main methods The activity of three diets (hyperlipidic with 6% fats; hypolipidic with 0.1% fats; and purified with 4% fats) was tested in CD1 mouse model experimentally infected with RML scrapie strain. Presence and distribution of typical central nervous system (CNS) lesions and deposits of PrP sc were evaluated by histopathological analysis and immunohistochemistry. Analysis of lipids was performed in homogenate and insoluble brain fraction of the neuronal membrane rich in LR. Key findings Results show that a diet with a different lipid level has not a significant role in the development of the scrapie disease. All infected mice fed with different diets died in the same time span. Histology, immunohistochemistry, and neuropathological analyses of the infected brains did not show significant differences between animals subjected to different diets. Significance Independently of the diet, the infection induced a significant modification of the lipid composition in homogenates, and a less noticeable one in insoluble brain fraction.

UNLABELLED The aim of this work was to determine the chemical constituents and in vitro antimicrobial activity of the essential oil (EO) of the aerial parts of Mentha sueveolens spp. insularis grown in Sardinia (Italy) against probiotic and starter microorganisms. The gas chromatography-mass spectrometry (GC-MS) analysis allowed to identified 34 compounds, most of oxygenated monoterpene compounds (82.5%) and among them, pulegone was found as major compound (46.5%). The agar diffusion test carried out employing the EO of Mentha suaveolens spp. insularis showed a low antibacterial activity, in particular no action was noticed for probiotic bacteria belonging to lactic acid bacteria groups, whereas almost all yeasts strains tested were inhibited. The automated microtitter dilution assay showed a clear effect at increasing concentration of EO on the specific growth rate (μ) and extension of the lag phase (λ) only for S. xylosus SA23 among bacteria and for Saccharomyces cerevisiae, Tetrapisispora phaffii CBS 4417, Metschnikowia pulcherrima, and Candida zemplinina among yeasts. Results obtained in this work allow us to broaden the knowledge on the effect of EOs on probiotic and food-related microorganisms. PRACTICAL APPLICATION Mentha suaveolens spp. insularis may be used in combination with probiotic bacteria into the food matrix or encapsulated in coating and edible films for food preservation.

Aims: Oxidative stress is an imbalance in the pro-oxidant/antioxidant homeostasis, characterized by excess accumulation of reactive oxygen/nitrogen species (ROS/RNS) and free radicals that can be toxic for cells by initiating disruptive peroxidation reactions on cellular substrates such as proteins, lipids, and nucleic acids. Neurons have a high content of unsaturated fatty acids which are easily peroxidable by the elevated levels of ROS and RNS produced by brain oxygen metabolism, yielding isoprostanes among which 8-iso-PGF2α derived from arachidonic acid represents a stable marker of lipoperoxidation, in vivo. Numerous findings pointed to the protective role of natural products against oxidative stress in the brain. Methodology: In the present work we evaluated the effects of a natural formula containing bacopa extract, vitamin E, astaxanthin and phosphatidylserine on lipoperoxidation in rat brain cortex, both in vivo and in vitro. Results: The results demonstrate that the natural formula could reduce basal and hydrogen peroxide- and amyloid β peptide-induced oxidative stress, as evidenced by the reduction of 8-iso-PFG2α and ROS/RNS production in the rat brain. Conclusion: Results could account for a rational use of herbal products in the treatment of conditions characterized by increased burden of oxidative stress and defective antioxidant mechanisms, such as aging and neurodegenerative disorders.

Aims: Sideritisitalicais a medicinal plant used for medical purposes mainly based on experiences rather than scientific evidence. Biological properties, composition of primary and secondary metabolites as well as the antioxidant capacity were investigated on samples from wild plant. Methodology: The ultrastructure of aerial parts and quantitative distribution of pigments, including chlorophylls and amino acids, as well as the main class of secondary metabolites (phenols, flavonoids, flavonols and proanthocyanidins) wer e investigated. The extracts were tested by radical scavenging assays (DPPH, ABTS) and pharmacological assays (antiproliferative activity, effects on ROS production and protective effects against DNA damage induced by hydrogen peroxide) for their effects o

Seeds of Tagetes lucida were imported to Egypt from Canada and propagated under greenhouse conditions in peat moss media. Soil was sandy in texture and the irrigation system was dripping irrigation. The growth parameters were determined at five successive plant ages, fresh and dry weights of herb were determined at three successive plant ages. The yield of aerial parts after 175 days, was about 7.5 Mg/ha. The essential oil (EO) was extracted by hydro-distillation for three hours with a yield of about 0.5% (w/v). The EO of each sample was subjected to gas-chromatography/mass spectrometry analyses to study the chemical composition. The main component of the EO was identified as methyl chavicol which matched over 90% of the whole composition. Chlorophyll a and carotenes increased with increasing plant age in both sites and seasons. Flavonoids decreased with the development of plant age, while the opposite was true with coumarines content.

The chemical composition of essential oil obtained by hydrodistillation of the fresh aerial parts of Mentha requienii Bentham (Lamiaceae) collected on the Gennargentu Mountains (Sardinia, Italy) has been investigated by gas chromatography and gas chromatography-mass spectrometry. The main constituents that resulted were pulegone (78%), menthone (0.5%), isomenthone (18%), isopulegone (1.3%) and limonene (1.76%). In vitro antifungal activity is evaluated in order to identify new means that could be helpful in the prevention of contamination in indoor environments.

Cannabinoids, endogenous and exogenously administered, are known to positively regulate food intake and energy balance. Since CB1 receptor antagonists reduce food intake and antagonize overweight, we developed a new CB1 receptor antagonist in an attempt to identify a compound with potential application in overeating disorders. The newly developed SM-11 compound dose-dependently decreases food intake in rats by 15–20%. Moreover, SM-11 reduces self-administration of palatable food in both food restricted and ad libitum fed rats, suggesting an action on the hedonic component of food intake. Thus, we next tested the effect of SM-11 on the stimulating properties of the CB1 receptor agonist WIN55,212-2 (WIN) on the electrophysiological activity of Nucleus Accumbens-projecting dopaminergic neurons of the ventral tegmental area (VTA). SM-11 fully and readily antagonized the WIN-induced increments in single spiking and burst firing of antidromically-identified dopamine neurons. When administered to naive (no WIN-pretreated) rats, SM-11 did not alter basal neuronal activity, thereby suggesting a pure antagonistic profile. SM-11 thus appears as a promising candidate in the search of potential anti-obesity medications.

We investigated the degradation of chlorinated phenols under reductive electron transfer reaction conditions. Although Li and Na metal proved useless, activated forms of these metals, either their soluble naphthalene radical anions or 1,2-diarylethane dianions, promoted the degradation of the starting materials to various extents. Additionally, efficient dehalogenation of the sodium salts of several mono-, di-, and tri-chlorophenols was obtained by their reduction with an excess of Na metal and a catalytic amount of naphthalene. .

The components of pompia, a hybrid Citrus species cultivated only in Sardinia (Italy), were extracted using an environmentally-friendly method and food-grade solvents. Taking into account that only few data are available on pompia composition, the phytochemical fingerprint of its rind extract was obtained by accurate component separation and identification, combining HPLC and mass spectrometry. Different flavones such as naringin (23.77 mu g/mg), neoeriocitrin (46.53 mu g/mg) and neohesperidin (44.57 mu g/mg) were identified. Additionally, the antioxidant activity and phenolic content were confirmed by DPPH and Folin-Ciocalteu assays. The whole extract was incorporated in innovative phospholipid vesicles, namely glycerosomes, hyalurosomes and glycerol containing hyalurosomes, which were prepared using a high ratio of extract/phospholipid (1/3.5 w/w). The in vitro biocompatibility of the nanoincorporated extract and its ability to potentiate the aptitude of the extract to counteract oxidative stress in skin cells were evaluated. The vesicles, especially glycerol containing hyalurosomes, were able to prevent oxidative damage and death of both keratinocytes and fibroblasts, promoting their viability. (C) 2016 Elsevier B.V. All rights reserved.

Seven commercial Moroccan honeys were considered for chemical characterisation. Volatile fraction, total polyphenols content, antioxidant and antiradical activities were evaluated by employing different analytical methodologies. Several physical parameters such as refractive index, pH, water content, solids content and colour were measured. Volatile fraction revealed an abundant presence of cis- and trans-linalool oxide in the seven studied samples. The presence of high levels of compounds related to the Maillard reaction, like furfural and hydroxymethylfurfural, could be the result of thermal treatments used to liquefy commercial honeys or of long storage times. The CIE L*a*b*Cab*hab° chromatic coordinates confirmed the advanced stage of the Maillard reaction, showing L* values lower than the common values found for honey of similar typologies.

A new combined “untargeted” and “targeted” metabolomics approach to investigate Rosmarinus officinalis L. plants was proposed by using the merging of high-performance thin-layer chromatography (HPTLC) assay and liquid chromatography mass spectrometry (LC–MS) with principal component analysis (PCA). Rapid fingerprints of Rosmarinus extracts were obtained and compared by using HPTLC technique; targeted compounds were identified and quantified by LC–MS/MS (MRM), while the antioxidant capacity of extracts was evaluated by methods based on the ability to scavenge radicals such as ABTS+* and DPPH*, since the metabolite expression is generally correlated with the biological activity. By HPTLC fingerprints, a rapid visual comparison of secondary metabolites in Rosmarinus samples from different places of collection was obtained and PCA was performed providing an overview of the capacity of the HPTLC variables to discriminate samples. HPTLC fingerprint assisted by PCA results a reliable untargeted approach for the discrimination and classification of different samples on the basis of their geographical area, from west to east of Sardinia. The ‘targeted’ approach was performed by developing LC–MS/MS (MRM) method allowing the identification and quantification of 12 compounds belonging to the family of flavonoids and phenolic acids. The combination of quantitative data with PCA allowed to discriminate La Maddalena Rosmarinus sample from others, in a correlated way with antioxidant activities results, showing values significantly higher. A positive Pearson correlation between antioxidant activity measured with DPPH and ABTS and total phenolic contents, finally confirmed that antioxidant activity depends on phenolic amounts.

High concentrations of human non pancreatic secretory phospholipase A2 (hnps-PLA2) have been reported as inducing factors in different inflammatory diseases. Thus, hnps-PLA2 inhibitors would be potential drugs against disorders generated by high levels of this enzyme. The latter has been crystallized with different ligands and several classes of inhibitors are known, but the optimization of their therapeutic properties requires: (i) a better understanding of the inhibitorprotein interaction mechanism, and (ii) finding a strategy to predict the activity of new molecules. Approaches related to computational chemistry may help to resolve these two problems. An automated docking study was performed on a series of 188 competitive hnps-PLA2 inhibitors. The docking data were then used to establish 3D QSAR models by combining Comparative Molecular Field Analysis (CoMFA) and PLS modeling. The robustness and prediction power of the best model were assessed with help of cross-validation and test set procedures that delivered excellent scores. The search for the best inhibitors against hnps-PLA2 has to be associated with a high specificity of the molecules selected, minimizing possible human side effects. This requires keeping at an extremely low level the inhibitors activity against human pancreatic phospholipase A2 (hp-PLA2) which is in negligible concentration in all tissues except in pancreatic ones. Then, the above mentioned modeling procedure was applied also on a series of hp-PLA2 inhibitors and, once more, the 3D QSAR model thus generated showed an excellent robustness and prediction power. Finally, the combination of the two models generated on hnps-PLA2 and hp-PLA2 offered a global predictive tool able to select new strong anti-inflammatory drugs with negligible side effects, at least at pancreatic level.

Recent improvements in many computational procedures able to establish predictive biological models, reinforce the necessity to introduce virtual High Throughput Screening (v-HTS) strategies in the early steps of the Drug Discovery process for a better rationalization. More particularly, Genetic Algorithms, Artificial Neural Networks and Fuzzy Logic methods seem to be the most promising tools to speed up and optimize the search for new leads and focused libraries. However, whatever method is used, real industrial applications need to establish well-defined and widely accepted protocols for validating the models and defining their robustness, prediction power and applicability domain.

An adaptive fuzzy partition (AFP) algorithm was applied on two bioavailability data sets subdivided into four ranges of activity. A large set of molecular descriptors was tested and the most relevant parameters were selected with help of a procedure based on genetic algorithm concepts and stepwise method. After building several AFP models on a training set, the best ones were able to predict correctly 75% of the validation set compounds. Furthermore, an improvement of about 15% in the validation results was got, on the same data set, as regard to other prediction methods. The importance to work with data sets including a large molecular diversity, and to use tools able to manage it, was also shown. The prediction power was increased up to 25% employing a data set with a better-optimised molecular diversity.

The essential oils from nine wild populations of the Sardinian Thymus herba-barona Loisel (Labiatae) were chemically characterized to identify the different chemotypes. GC and GC–MS analysis showed that in Sardinia only two chemotypes (thymol and carvacrol) are present. The carvacrol chemotype is predominant and this compound was found in high concentration (50.7–79.4%) in seven of the nine populations studied. The thymol chemotype was found in only two populations (34.2–56.2%). Analyses also showed that the other main components of the oils of T. herba-barona, were 3-heptanone (1.4–5.6%), 3-octanone (1.4–5.5%) and borneol (2.5–8.3%). p-Cymene is present at 1.2–5.6% but is not present in plants from the Tonara medium station, where O-cymene (1.9%) is present instead. Copyright © 2002 John Wiley & Sons, Ltd.

Ethanol extracts of Stachys glutinosa L. (Lamiaceae) were investigated for antioxidative properties, as well as antiproliferative action on various cell lines. The antioxidant activities were investigated by ABTS (2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid) assay, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, β-carotene/linoleic acid assay, scavenging of hydrogen peroxide (horseradish peroxidase test), superoxide anion scavenging, and hypochlorous acid scavenging (taurine test). The antioxidant activity was reported as IC50 and reveals antioxidant effects. Antiproliferative effects were measured in vitro on three cell lines: HepG2 (human hepatocarcinoma), MCF7 (breast human adenocarcinoma) and C2C12 (mouse myoblast) cell lines by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The ethanol extract induced variations in cell viability on all cell lines tested. At 200 μg/mL, the effects on cell viability were - 23%, - 27% and - 37%, respectively, for C2C12, MCF7 and HepG2.