(SIF), in Calenda- Rio Dal 28 Al 30 Giugno a Pisa, Presso La Sala Convegni – Polo Piagge Dell’Univer- Sità, in Via Matteotti, 11

dizione numero quindici per il congresso della Società Italiana Edi Fitochimica (SIF), in calenda- rio dal 28 al 30 giugno a Pisa, presso la Sala Convegni – Polo Piagge dell’Univer- sità, in via Matteotti, 11. Il XV Congresso Nazionale della SIF si svolge in contemporanea con il 1° Congresso Internazionale sulla piante commestibili, medicinali e aromatiche (1st International Congress on Edible, Medicinal and Aromatic Plants - ICE- MAP 2017). Il Congresso ha inizio nel pomeriggio di mercoledì 28 giugno con due conferenze plenarie e continuerà il giovedi 29 e il venerdì 30 con ulteriori conferenze plenarie, letture brevi, presentazioni di poster e la tavola rotonda sulle più re- centi questioni relative ai botanicals. La lingua ufficiale è l’inglese e non è prevista alcuna traduzione simultanea delle varie relazioni.

Ecco in sintesi gli argomenti affrontati nel corso del Congresso. - Plants and the environment, biodiversity and conservation. - Edible, medicinal and aromatic plants: from the field to the final product. - Isolation and structural elucidation of natural products and their bioactivity. - Edible plants with health‐promoting properties. - Plant biotechnology. - Synthetic approach to natural products. - Production and formulation technology of herbal products, botanicals and cosmetics. - Plants and natural products in veteri- nary medicine.

56 t natural 1 giugno 2017 INVITED SPEAKER INVITED SPEAKER TITLE Bucci M. Enrico Research Integrity and Misconduct: General Considerations and Notable Cases in Plant Biology Choi Young Hae Natural deep eutectic solvents: a potential media for natural products research Colombini Maria Perla Macromolecules in archaeological findings to disclose ancient secrets and daily life Efferth Thomas Microarray-based profiling of natural product inhibitors of NF-κB Peruzzi Lorenzo The role of chemosystematics in biodiversity studies

COMUNICAZIONI ORALI CONTRIBUTOR TITLE Argentieri Maria Pia et al Chemistry and biodiversity of genus Achillea Ascrizzi Roberta et al Wild harenna coffee: flavor profiling from the cherry to the cup Bottone Alfredo et al Flavonoids and terpenes from the leaves of the Italian almond varieties “Mandorla di Toritto” and “Mandorla d’Avola” Brighenti Virginia et al Metabolite profiling of anthocyanins inVaccinium myrtillus L. berries and food products Cosci Francesca et al Bioactivity of Cinnamomum verum essential oil against Varroa destructor, parasitic mite of Apis mellifera D’Ambola Massimiliano et al Antibacterial activity of constituents of Psiadia punctulata DC (Vatke) (Asteraceae) El Gendy AG Nasser et al Growth, Yield and chemical composition of essential oil of Mentha piperita var. multymentha grown under different agro-ecological locations in Egypt Giacomelli Chiara et al Effect of Ribes nigrum bud extract on human gingival mesenchymal stem cell cytokine release: a role on regenerative processes Giuliani Claudia et al Volatile profile of perennial leaves of seven cultivated species of Ceratozamia (cycads) Jameel Bzour Immunomodulatory potential of the Mediterranean sage (Salvia fruticosa) in adjuvant-induced arthritis in rats Lucarini Elena et al Tuscan black kale as source of compounds active on chemotherapy-in duced neuropathic pain: a hypothesis about the mechanism of action Maggi Filippo et al Evaluation of Hypericum androsaemum L. red fruit effects in a mouse model of post-stroke depression Marrelli Mariangela et al Echinophora tenuifolia L. aerial parts: phytochemical constituents and antiproliferative activity against human cancer cell lines Petretto Giacomo et al Volatile organic compounds and antioxidant activity of non-polar extract from Saffron stamens harvested in Morocco Piazzini Vieri et al Solid lipid nanoparticles for oral delivery of silibinin: formulation, characterization and in vitro evaluation using pampa and caco-2 cell models Pozzo Luisa et al Chemical characterization of blackthron (Prunus spinosa) fruit and evaluation of its protective effect against hepatic steatosis in rats Rigano Daniela et al Phytochemical investigation on the Northern African endemic plant Daucus virgatus Sgorbini Barbara et al Artemisia umbelliformis Lam. and génépi liqueur. From the field to the final product: the importance of the volatile fraction in quality control Smeriglio Antonella et al Polyphenolic profile and fatty acids composition of seed oils and their byproducts from Cannabis sativa L. varieties with potential health-promoting properties Sylwester S´ lusarczyk et al Practical aspects of using metabolomic approach in medicinal plants analysis focusing on roots from aeroponics. Taddeo Vito Alessandro et al HPLC investigation of the nutraceutical composition of the edible herb Amaranthus retroflexus L. Tardugno Roberta et al Phytochemical analysis of Cichorium intybus L. varieties from Veneto (Italy) region by means of HPLC-UV/DAD and HPLC-MS/MS Testai Lara et al The Citrus flavonoid, naringenin, protects the myocardium against ageing. Vassallo Antonio et al in-cells antioxidant potential of herbal preparations and components from Galactites elegans

giugno 2017 natural 1 t 57 ABSTRACT

Research Integrity and Miscon- Plant Biology is not exempt; while one could duct: General Considerations think that most of the dishonest behaviour would and Notable Cases be limited to those research fields where larger in Plant Biology funding is available, such as biomedical sciences, in facts some of the most notable cases of recent Enrico M. Bucci scientific misconduct are to be found among people pretending to study plant molecular biology. I think Sbarro Health Research Organization, Temple University, that all of us should know what is going on – what Philadelphia, U.S.A. are we risking, who are the fraudsters, what we can [email protected] do and what responsibility every single citizen has in knowing what Science really is – and what is not. When I started my activity in biomedical data analysis, I was fully convinced that Science could cor- Macromolecules in rect itself – and researchers as a community were archaeolo-gical findings motivated into telling the truth, notwithstanding to disclose ancient secrets some notable exception. In Science, every research- and daily life er builds upon trusted results obtained by others; which was exactly what I wanted to do, albeit on Maria Perla Colombini a scale larger than usual – aggregating the full biomedical literature in a single database, crossing data Dipartimento di Chimica e Chimica Industriale, University of from different sources and finding new potential an- Pisa Istituto per la Conservazione e Valorizzazione dei Beni Culturali swers to diverse scientific problems using the pow- – CNR er of informatics to get it quick and easy. Trusting [email protected] each other experimental findings is the usual habit Since ancient times, a wide variety of natural or- of researchers in every discipline – and I used to ganic materials have been used as adhesives, seal- perceive errors in Science as honest or due to slop- ants, painting and coating materials. Proteins, oils, py science, not to some bad and concealed purpose. gums, natural resins and resinous materials played It then came as a shock to me, when with my small a prominent role, since their intrinsic properties group and armed with the proper technology, we meant that they could be used not only as paint- discovered that, far from being an exception, the ing materials, adhesives, hydro-repellents, coating three cardinal sins of scientific fraud – data and ex- and sealing agents, but also as flavours, incense, in- periment fabrication, falsification and plagiarism – gredients for cosmetics, medicines and mummifica- are largely diffuse in the current scientific record, at tion balms. The chemical characterization of such such a rate that scientific fraud is becoming a seri- organic materials when properly integrated with ous issue. I started writing to scientific journals, to related information from historical sources and uncover potential frauds, ending up into involving archaeological data, has in the last few years con- the police for investigating some serious case. siderably improved our knowledge of painting tech- That because Science is not simply a cultural ex- niques, crafts and technologies of the past, and has ercise; it must help all of us in discriminating facts provided archaeologists with valuable information. from fictions and in making the right predictions, In fact, identifying specific materials from molecu- not to mention its role in the technological develop- lar patterns helps in assessing the role that these ment and in the general progress of humanity. The substances played and in determining the use of ar- human society as a whole, cannot allow the poison- tefacts on which these residues survive. The characterisation of organic materials in archae- ing of the very essence of Science – i.e. the unbiased ology is a challenge for a chemist, due not only to collection of facts to be screened with a method able the complexity of the chemical composition of the to discriminate whether a given hypothesis is true natural substances that may be present alone and or not. in mixtures, but also to changes in the chemical While fraud in science is not a new thing nor un- composition as a consequence of human activities. expected, given the human nature of scientists, the For instance, heating processes applied to plant scale at which today is occurring is previously un- resins deeply modified their chemical composition, seen – simply because there are too many incentives inducing aromatisation, demethylation and decar- and very little risks given the current organization boxilation reactions with formation of new species. of the scientific enterprise. Moreover, degradation due to ageing under the in- The result? More and more dishonest researchers fluence of different environmental circumstances are ruining Science – and if we will allow this, it will or improper storage leads to further changes in the be very hard to step back. composition of the original materials. This lecture

58 t natural 1 giugno 2017 outlines the analytical procedures based on mass verse medicinal plants to characterize their NF-κB spectrometric techniques applied to archaeological inhibitory activity. These plants were Daedalea gib- samples. In particular, direct exposure mass spec- bosa, Eleutherococcus senticosus, Rhodiola rosea, trometry (DEMS), pyrolysis with in situ derivatisa- Salvia miltiorrhiza, Salvia officinalis, Schisan- tion –gas chromatography/mass spectrometry (Py- dra chinensis, Scrophularia ningpoensis, Taxus GC/MS), and gas chromatography/mass spectrom- chinenis, and Ziziphus spina-christi. Furthermore, etry (GC/MS) will be discussed highlighting their we analyzed a number of isolated phytochemicals abilities in solving complex molecular mixtures and from these and other plants for their their NF- in achieving reliable results. κB inhibition, including 6’’’ feruloylspinosin, 6’’’ Basically, the identification of materials is based on sinapoylspinosin, 7-xylosyl-10-deacetylpaclitaxel, the detection of both molecular fingerprint and the artesunate, casticin, epigallocatechin, kaempferol, presence of one or more specific molecular biomark- pomolic acid, rosmarinic acid, scopoletin, spinosin, ers which have survived ageing or which are formed ursolic acid. A wide array of different methods and over the centuries as stable products of ageing pro- technologies have been applied to identify the mo- cesses. Case studies discovering materials used in lecular mechanisms of NF-κB inhibition by these the past as ointments and balms found in vessels of herbal extracts and isolated phytochemicals, such the 3th – 1th century B.C., and as jewels by South as GFP-basded reporter cell assays, Western blot- Italy people in the 7th century B.C. are presented. ting of cytosolic and nuclear protein fractions, flow cytometric detection of apoptosis, electromobility shift assay (EMSA), transcriptome-wide microarray Microarray-based profiling of hybridization, gene promoter binding motif analy- natural product inhibitors of sis, molecular docking and others. In the present NF-κB contribution we give an overview of the results and implications obtained. One of the NF-κB inhibitors, Thomas Efferth artesunate, has not only been tested in vitro and in vivo, but also in clinical phase I/II trials with human Department of Pharmaceutical Biology, Johannes Gutenberg cervical cancer and colorectal cancer patients. In University, Staudinger Weg 5, 55128, Mainz, Germany. conclusion, natural products and medicinal herbs [email protected] represent attractive tools for the targeted NF-κB in- Since the early days of cancer chemotherapy, the hibition. If such inhibitors are combined with stan- development of resistance was persisting as an un- dard chemotherapy, improved treatment responses resolved major impediment for treatment success. may be achieved resulting in prolonged survival of Drug resistance is multi-factorial in nature. Among cancer patients. the numerous resistant phenotypes is resistance to References: apoptosis. One mayjor player regulating apoptosis Efferth, Semin Cancer Biol. 2017 (epub ahead of print) is the transcription factor NF-κB (nuclear factor Seo et al., Molecules. 2016;21(4):496. kappa-light-chain-enhancer of activated B cells). Kadioglu et al., Phytomedicine. 2016;23(3):293-306. Its aberrant activation causes both carcinogenesis Hutterer et al., Antiviral Res. 2015;124:101-9. Wu et al., J Ethnopharmacol. 2015;176:55-68. and drug resistance, because cells compromised by Kadioglu et al., Anticancer Res. 2015;35(5):2645-50. carcinogenes or chemotherapeutic drugs cannot die Kadioglu and Efferth, J Nat Prod. 2015;78(4):762-75. but continue to grow, which ultimately leads to ag- Panossian et al., Front Neurosci. 2013;7:16. gressive malignant tumors. NF- B forms complex- Righeschi et al., Cancer Genomics Proteomics. 2012;9(3):143- κ 51. es with IκB (inhibitor of NF-κB) in the cytoplasm. Shen et al., Pharmaceuticals. 2012;5(2):189-208. Upon appropriate stimuli, proteasomal degradation Bachmeier et al., PLoS One. 2011;6(5):e20550 of IκB takes place followed by IKK (IκB kinase)-me- Ruimi et al., Int J Mol Med. 2010;25(3):421-32. diated phosphorylation and nuclear NF-κB translo- Jiang et al., Int J Oncol. 2008;33(1):103-11. Efferth et al. J Mol Med. 2002;80(4):233-42. cation. NF-κB induces the expression the expression of anti-apoptotic genes that cause drug resistance. Therefore NF-κB along with its signal transduction The role of chemosystematics partners has been recognized as promising target in biodiversity studies for therapeutic interventions. Many synthetic small molecules, peptides, antisense RNAs and gene ther- Lorenzo Peruzzi, apy strategies, but also natural compounds have Dipartimento di Biologia, Università di Pisa, Via Derna 1, been proposed as NF-κΒ-targeted approaches. The 56126 Pisa, Italy; [email protected] concept is to inhibit NF-κB to lower the threshold for apoptosis induction and thereby to re-sensitize The comparative study of chemical profiles in plants tumor cells to standard chemotherapy. Over the can provide a high number of taxonomic characters, past years, we investigated herbal extracts from di- useful to test their congruence with existing clas-

giugno 2017 natural 1 t 59 ABSTRACT

sifications, either at macro- or micro-evolutionary Many scientific questions could be explained by the scale, and to identify peculiar chemotypes. In ad- hypothesis of the presence of alternative media in dition, the study of floral bouquets and other sec- nature, e.g. low-solubility of natural products, mys- ondary metabolites represent a crucial contribution tery of biosynthesis, transportation, and storage of to the knowledge on pollination ecology, on inter- non-water-soluble metabolites, and so on. Based on action of plants with herbivores/pathogens, and on these questions combined with ILs or DES mecha- allelopathy. A brief overview of the potentialities of nisms, we have suggested that in nature there should chemosystematics in current biodiversity studies is be a wide range of ILs or DES to play diverse roles, presented, based on selected case studies from gym- called “Natural Deep Eutectic Solvents” (NADES) nosperms (e.g. Juniperus), monocots (e.g. Crocus), as a common term for these mixtures, ILs or DES eudicots (e.g. Lavandula, Polygala, Stachys). whose ingredients originate from nature. Incidentally, this kind of investigation can also have As the first step to prove this, we made various com- “secondary outcomes” very useful to human health, binations of these candidates, thereby discovering concerning for instance the discovery of new natural more than 150 combinations that form viscous liq- products, often showing bioactivity. Accordingly, an uids. In this presentation, I would like to show di- ever closer collaboration among taxonomists, con- verse combinations of NADES and their potential ap- servationists and phytochemists is expected for the plications, particularly for natural products research future, in order to push forward the knowledge of the and current problems of the liquids for practical ap- plant world in a synergistic manner. proach.

Natural deep eutectic solvents: References: 1. Choi Y.H. van Spronsen J., Dai Y., Verberne M., Hollmann F., a potential media for natural Arends I.W.C.E., Witkamp G.-J, Verpoorte R. Plant Physiol. 156, products research 1701-1705, 2011. 2. Dai Y., van Spronsen J., Witkamp G. -J., Verpoorte G.-J., Choi Young Hae Choi, Yuntao Dai, Hennivanda, Natali Y.H. Anal. Chim. Acta 766, 61-68, 2013. Rianika Mustafa, Hye Kyong Kim, Robert Verpoorte

Natural Products Laboratory, Institute of Biology Leiden University, Sylviusweg 72, 2333 BE Leiden, the Netherlands. [email protected] Mixing salts and/or organic compounds may cause a considerable reduction of the melting point, turn- ing them into liquids even at very low temperatures. The field of ionic liquids (ILs) starts in the beginning of 20th century reported on the physical properties of ethylammonium nitrate. But it is only in recent years that ILs and deep eutectic solvents (DES) have been revisited by chemical engineering to utilize them as green media for chemical processing. Using the liquids made from synthetic chemicals ILs and DES now have many different applications such as dissolving polymers and metals, and media for bio- Achillea millefolium transformation. Although ILs and DES are considered as one of the most promising media for green processing, the ingredients, mostly from chemical Chemistry and biodiversity of synthesis are usually toxic both to human and en- genus Achillea vironments. This hurdle is still to surpass for the a b a utilization of the solvents. In analogy with the syn- Argentieri Maria Pia , Vitalini Sara , Avato Pinarosa thetic ILs our group has recently hypothesized that a the metabolites that occur in large amounts in cells Dipartimento di Farmacia-Scienze del Farmaco, Università degli Studi “Aldo Moro” di Bari, Via Orabona 4, 70125 Bari, Italy. may form a third type of liquid – one separate from bDipartimento di Scienze Agrarie ed Ambientali, Università degli water and lipids (1, 2). Studi di Milano, via G. Celoria 2, 20133 Milano, Italy. Taking the plant metabolomics data, which we have [email protected] collected over past years into consideration we saw a clear parallel with the synthetic ILs. This discov- The rich biodiversity of the Alps offers the opportu- ery could break down a long-believed fundamental nity to explore different botanical species that were theory that water is the unique media in nature. used as traditional remedies in the various valleys.

60 t natural 1 giugno 2017 In this contest we have collected in the Valtellina areas plants belonging to the genus Achillea (A. millefolium L. and A. moschata Wulfen). The genus Achillea (Asteraceae) includes about 130 flowering and perennial species mostly distributed in Europe and temperate areas of Asia. The species belonging to this genus have been used since ancient time for the treatment of wounds, bleeding, headache, inflammation, pains, flatu- lence, dyspepsia. Phytochemical investigations of Achillea species have revealed that many secondary metabolites, such as flavonoids, terpenoids, amino acids derivatives and fatty acids, are highly bioactive. Within the genus, A. millefolium is the most studied and it is the only species listed in the European Pharmacopoeia. Its water infusion or alcohol extract is widely used in Europe as a remedy to treat digestive problems, diabetes, hepato-biliary disease and amenorrhea, and also consumed for its Drupe di caffè antitumor, antimicrobial, anti-inflammatory and antioxidant properties1. Flavonoids (apigenin and Wild harenna coffee: flavor quercetin) and phenolic acids (caffeoylquinic acids) profiling from the cherry to are reported to be the major compounds present the cup in A. millefolium and they are the responsible of Roberta Ascrizzi1, Guido Flamini1,2 the antioxidant activity reported for this species2. In view of the important applications of A. millefo- 1 Department of Pharmacy, University of Pisa, Via Bonanno 6, lium for human health, in this work we have stud- 56126, Pisa, Italy 2 ied the phytochemical diversity of A. millefolium Centro Interdipartimentale di Ricerca “Nutraceutica e Alimentazione per la Salute”, Università di Pisa, Italy and A. moschata collected in different geographi- [email protected] cal areas. A. moschata grows on the Alps where it is extensively used by local people for its medici- Coffee is one of the world’s most popular beverag- nal properties as reported in recent ethnobotani- es, representing a major source of income for many cal studies3. Our research aimed to illustrate the producing countries. The typical aroma is certainly phenolic compositional profile of the aerial parts of the reason of its success: it depends on the coffee these two species. To the best of our knowledge, variety, with its specific set of chemical precursors, no phytochemical and biological reports have been as well as on the processing and consumption phases published previously for A. moschata. [1]. In the Harenna Forest, in the Bale National Park Data showed a different metabolic profile between of Ethiopia, at an altitude of around 1800 meters, an the two species. A. moschata is richest in phenolic Arabica wild coffee grows spontaneously: the Haren- acids than A. millefolium. Moreover in this work na Forest Wild Coffee Presidium recognizes the val- we have observed phytochemical diversity within ue of this unique and high quality product, which has the same species. In fact the methanolic extracts of been bestowed the Slow Food Presidium in 2012. It A. moschata are characterized by the same meta- is naturally dried in loco and its exports volumes are bolic profile but they differed in the amount of main limited, as its availability is greatly variable. compounds. The aroma of coffee is a complex bouquet of odor- ants of different origin: the evaluation of its composi- 1 Dias MI., Barros L., Duenas M., Pereira E., Carvalho AN., Alves RC., Oliveira MBPP., Santos-Buelga C., Ferreira ICFR. Chemi- tion could be used as a tool for the coffee industry to cal composition of wild and commercial Achillea millefolium L. control desirable flavor outcomes of coffee through and bioactivity of the methanolic extract, infusion and decoc- processing or other farm management technique. In tion. Food Chem 2013, 141, 4152-4160. this perspective, the Head-Space analysis could rep- 2 Benedek B., Gjoncaj N., Sukel J., Kopp B. Distribution of phenolic compounds in Middleeuropean taxa of the Achillea millefo- resent a simple, fast and accurate technique to eval- lium L. aggregate. Chemistry & Biodiversity 2007, 4, 849-857. uate the development of flavor-active compounds in 3 Vitalini S, Puricelli C., Mikerezi M., Iriti M. Plants, people and all the phases of the coffee production. tradition: ethnobotanical survey in the Lombard Stelvio Nation- In the present study, the volatile aroma profile of Ha- al Park and neighbouring areas (Central Alps, Italy). J. Ethno- pharmacol 2015, 15(173), 435-458. renna wild coffee has been analysed by means of HS-

giugno 2017 natural 1 t 61 ABSTRACT

SPME-GC/MS from the coffee cherry to the coffee Flavonoids and terpenes from cup, all throughout the processing phases which lead the leaves of the Italian almond to the final product. The dried green cherries, rich in varieties “Mandorla di Toritto” monoterpenes (especially dihydro myrcenol and lim- and “Mandorla d’Avola” onene), esters (mainly cis-2-tert-butyl-cyclohexanol acetate), alkanes, and phenylpropanoids, also con- Alfredo Bottone, Paola Montoro, Milena Masullo, tain the precursors for the aroma compounds for- Cosimo Pizza, Sonia Piacente. mation through the Maillard reaction, which takes place during the roasting phase. Pyrazines (mainly Università degli Studi di Salerno, Dipartimento di Farmacia, 2,6-dimethylpyrazine, 2-ethyl-6-methylpyrazine, Via Giovanni Paolo II, 132 84084 Fisciano (SA) and 2,3,5-trimethyl pyrazine) begin to be detected [email protected] at 100 °C, to become the most abundant chemical Prunus dulcis (Rosaceae) is a tree native of Minor class on the later roasting phases, when the tempera- Asia, widely cultivated for its edible seeds. Much ture reaches 160, 180, and 204 °C, the final roasting is known about their chemical composition1,2 and temperature. Pyridines (mostly pyridine) are also their benefits for human health, mainly due to the developed during the roasting phase: they are only high content of polyunsaturated fatty acids3. On detected in the samples roasted over the tempera- the other hand, very few is known about the by- ture of 160 °C. Furans (mostly 2-furan methanol, products such as leaves, husks and shells. In the 5-methyl furfural and furfuryl acetate) are also de- frame of a project aimed at defining the metabolome veloped through thermal treatment, while monoter- of the different parts of the Italian varieties of penes decrement as they are volatilized very soon in Prunus dulcis, the leaves of the variety “Mandorla the roasting process. This is also confirmed by their di Toritto” have been investigated. Final target detection in the coffee silverskins, for which the vol- is to valorize such biomasses deriving from the atiles emission profile is similar to the green cherries manufacturing processes as a source of bioactives one: they retain the early-released volatiles as they to employ for the production of nutraceutical and are soon separated from the cherries in the roaster. cosmetic formulations rich in polyphenols, in order Acknowledgment For the present project, we want to acknowledge to cut the high costs related to their disposal. The Mr. Andrea Trinci of Trinci Torrefazione Artigiana di Caffè e Cacao methanolic extract was first submitted to LC-MS (Cascine di Buti, Pisa, Italy): he freely provided us the samples for experiments to obtain a complete profile of the the analyses. There is no financial interest of Trinci Torrefazione secondary metabolites and succesively purified by Artigiana di Caffè e Cacao related to this project. size-exclusion chromatography and by RP-HPLC. [1] Yeretzian, C. Coffee. In Springer Handbook of Odor; Buettner, The structures of the isolated compounds were A; Springer International Publishing AG: Zurich, Switzerland, elucidated by NMR experiments, and confirmed 2017; pp 107-127. by MS experiments. More kinds of extracts were prepared by employing eco-friendly solvents and protocols, and compared by LC-MS experiments. The metabolic profile of the apulian leaves was compared with those of the sicilian varieties of the “Mandorla d’Avola” (Fascionello, Pizzuta and Romana) by LC-MS experiments, and processing the data by multivariate analysis. Furthermore, total phenolic content and radical scavenging activity of the extracts was evaluated. This study showed how the leaves of the apulian “Mandorla di Toritto” and the sicilian “Mandorla d’Avola” varieties are a rich source of phenolic compounds with antioxidant activity, suggesting their application in nutraceutical and cosmetic fields.

1) Milbury, P. E.; Chen, C. Y.; Dolnikowski, G. G.; Blumberg, J. B. Determination of flavonoids and phenolics and their distribution in almonds. J. Agric. Food Chem. 2006, 54, 5027– 5033. 2) Sathe, S. K.; Seeram, N. P.; Kshirsagar, H. H.; Heber, D.; Lapsley, K. A. Fatty acid composition of California grown almonds. J. Food Sci. 2008 , 73 , 607-614. Prunus amygdalus

62 t natural 1 giugno 2017 3) Berryman, C. E.; Preston, A. G.; Karmally, W.; Deckelbaum, in V. myrtillus food derivatives. Prior to analyses, R. J.; Kris-Etherton, P. M. Effects of almond consumption on anthocyanins were selectively extracted from the the reduction of LDL-cholesterol: a discussion of potential mechanisms and future research directions. Nutrition Reviews. samples of food products by means of dynamic 2011 , 69 , 171–185. maceration with acidified methanol as the extraction solvent. The analysis of anthocyanins was then carried out by means of RP-HPLC-UV/DAD and ESI-MS. As a comparison, Vaccinium myrtillus L. berries from wild crops collected in the Italian Northern Apennines were analyzed in parallel. The quali- and quantitative profile of anthocyanins in 3 samples of bilberry fruit, 14 bilberry juice, 14 bilberry liqueurs and 22 bilberry jams from the Italian Northern Appennines was evaluated in order to monitor their quality. In total 14 anthocyanins, comprising the 3-O glucosides, galactosides and arabinosides of the anthocyanidins delphinidin, cyanidin, petunidin, peonidin and malvidin, were detected in the fruit samples as well as in the derived food products considered in this study. The great majority of the analysed samples showed a high content of bioactive compounds, Vaccinium myrtillus thus highlighting the high biological values of the products. Metabolite profiling of The anthocyanin profiles of bilberry jams and juice anthocyanins in was comparable to those of the fruit, suggesting that Vaccinium myrtillus L. berries it wasn’t affected by the process of preparation. On and food products the other hand almost all liqueur samples exhibited a different quantitative profile of the anthocyanins Virginia Brighentia, Chiara Ranieria, Federica detected. a a Pellati , Stefania Benvenuti 1. Smeriglio, A.; Monteleone, D.; Trombetta, D. Health Effects of Vaccinium myrtillus L.: evaluation of efficacy and [email protected] technological strategies for preservation of active ingredients. Mini Rev. Med. Chem. 2014 , 14 , 567 - 584. aDepartment of Life Sciences, University of Modena 2. Smeriglio A.; Barreca, D.; Bellocco E.; Trombetta D. and Reggio Emilia, Chemistry, pharmacology and health benefits of anthocyanins. Via G. Campi 103, 41125 Modena, Italy. Phytother. Res. 2016 , 30 , 1265 - 1286. 3. Queiroz, F.; Oliveira, C.; Pinho, O.; M.P.L.V.O. Ferreira, I. Vaccinium myrtillus L. is a spontaneous plant Degradation of anthocyanins in blueberry jams/stuffed fish. J. Agric. Food Chem. 2009 , 57 , 10712 - 10717. native to the mountain areas of Northern and Central Europe. Its fruit (commonly known as bilberry) is widely employed in the traditional cuisine of this area 1,2. Indeed, several bilberry derivatives are available on the market, including juice, jams and spirits 3. Anthocyanins represent the most abundant class of bioactive compounds present in bilberry fruit, conferring it several health- promoting properties 1,2. However, the content of these secondary metabolites in food products can be affected by the processes of preparation, making the monitoring of the quali- and quantitative profile of these compounds a crucial point for their genuineness and quality assurance 2,3. In the light of all the above, this study was aimed at the determination of the anthocyanin profile Cinnamomum verum

giugno 2017 natural 1 t 63 ABSTRACT

Bioactivity of Cinnamomum bees without causing significant mortality of the verum essential oil against bees. In the field,C. verum EO was able to reduce Varroa destructor, parasitic V. destructor population up to about 70%. mite of Apis mellifera This study indicated that C. verum could be a promising repellent against V. destructor used as an Francesca Cosci1, Guido Flamini2, Roberta alternative to chemical treatment in integrated pest Ascrizzi2, Rossella Bocchino1, Jessica Girardi1, management strategies in bee colonies. Barbara Conti1, Stefano Bedini1* 1. Bedini, S.; Flamini, G.; Girardi, J.; Cosci, F.; Conti, B. Not just 1Department of Agriculture, Food and Environment, University for beer: evaluation of spent hops (Humulus lupulus L.) as of Pisa, Via del Borghetto 80, 56124 Pisa, Italy; a source of eco-friendly repellents for insect pests of stored 2Department of Pharmacy, University of Pisa, Via Bonanno 6, foods. J Pest Sci. 2015 , 88(3) , 583-592. 56126 Pisa, Italy. 2. Bedini, S.; Flamini, G.; Cosci, F.; Ascrizzi, R.; Echeverria, M. [email protected] C.; Guidi, L.; Landi, M.; Lucchi, A.; Conti, B. Artemisia spp. essential oils against the disease-carrying blowflyCalliphora vomitoria. Parasit Vector. 2017 , 10 (1) , 80 The honeybee, Apis mellifera (Hymenoptera Apidae) has a great economic and ecological importance. In fact, beside in the beehive products, A. mellifera plays a major role in the pollination of wild angiosperm and crops species, essential for ecosystem functioning and human food production. Varroa destructor (Acari Mesostigmata Varroidae), an ectoparasitic mite, is the most devastating parasite of the Western honeybees and causes substantial losses of colonies worldwide. Currently, the control of this pest mainly relies on chemicals such as oxalic acid, pyrethroids, and organophosphates. However,

the extensive and repeated use of acaricides Foto di Bart Wursten have caused the development of resistance in V. destructor populations, causing a reduction of their efficacy and are toxic to bees. Moreover, these products may leave residues in the hive, contaminating honey and beeswax. For these reasons, the use of alternative strategies for the control of V. destructor are required. Essential oils of aromatic plant are known to have Psiadia punctulata repellent, insecticidal and acaricidal properties (2) and could represent a very valuable low-cost, environmental-friendly alternative to synthetic Antibacterial activity of products (1). Cinnamomum verum (Lauraceae) J. constituents of Psiadia Presl is an evergreen tropical tree originated in Sri punctulata DC (Vatke) Lanka, traditionally widely used to flavour foods. (Asteraceae) C. verum bark yields an essential oil containing a b cinnamaldehyde and eugenol as main constituents Fabrizio Dal Piaz , Massimilano D’Ambola , Nicola a a a that showed important biological activities, such Malafronte , Anna Maria Petrone , Amalia Porta , c b as peripheral vasodilatory, antitumor, antifungal, Ammar Bader , Lorella Severino , Nunziatina De Tommasia*, Angela Bisiod cytotoxic and antimutagenic as well as insecticidal a and insect repellent activities. Dipartimeno di Farmacia, Università di Salerno, Fisciano, (SA), Italy. In this work, cinnamon bark essential oil was b Dipartimento di Medicina Veterinaria e Produzioni Animali, extracted, chemically analysed and tested to evaluate Università degli Studi di Napoli Federico II, via Delpino 1, its toxic/repellent activity against V. destructor in 80137, Naples, Italy; cDepartment of Pharmacognosy, Umm Al-Qura University, 21955 both laboratory and apiary. Results of laboratory Makkah, Saudi Arabia. tests showed that C. verum EO is able to cause, at dDipartimento di Farmacia, Università di Genova, Via Brigata the concentration of 2%, 24h after the treatment, Salerno 13, 16147 Genova, Italy. the detachment of 50% of the mites present on adult [email protected]

64 t natural 1 giugno 2017 Psiadia punctulata (DC) Vatke (Asteraceae) is small shrub with leaves, especially when young, covered by a gummy exudate. The decoction of the leaves of P. punctulata is traditionally used in Africa for the treatment of colds, fevers, abdominal pains and for the removal of ec- toparasites from cattle (1). P. punctulata is well Foto di Bart Wursten known in the ancient Arabic medicine. P. punctulata is distributed in Africa and tropical Asia (2), and included into the Euro+Med genera of Astereae (3). Chemical studies on P. punctulata extracts reported the presence of ent-kaurane and ent-trachylobane, diterpenes (1, 4), flavones and phenylpropenoids (5, 6) as main components. In this study, a bioassay-oriented fractionation approach of the surface extract of P. punctulata from Arabian region for evaluation of antimicrobial activity against Staphylococcus aureus, Pseudomo- nas aeruginosa and Candida albicans was carried out. the crude surface extract showed antibacteri- Psiadia punctulata al activity against S. aureus (MIC50 = 100 μg/mL, Growth, Yield and chemical MIC100 = 180 μg/mL), while no activity was de- composition of essential tected against P. aeruginosa. Moreover, the extract showed antifungal activity against C. albicans, with oil of Mentha piperita MIC2 = 50 μg/mL and MIC0 = 130 μg/mL. var. multymentha grown The extract was subsequently separated to afford under different agro- thirteen kaurane and trachylobane diterpenes along ecological locations in Egypt with nine flavones and seven diterpenes already described in the literature. Three flavonoids showed El Gendy A. G. a, Hendawy S. F.a , Omer E.A. a, antimicrobial activity (MIC50 > 80 μg/mL). Laura Pistellib and Luisa Pistellic All the diterpenes with exception of one were inac- tive at the concentrations tested. aDepartment of Medicinal and Aromatic Plants Research, National Research Center, 12622, Dokki, Giza, Egypt The ability of these compounds to inhibit the bio- bDepartment of Agriculture, Food and Environment, University of film formation ofC. albicans and S. aureus was Pisa, Pisa, Italy. then analysed. cDepartmentof Pharmacy, University of Pisa, Via Bonanno 33, The maximum antibacterial effect was shown by a 56126 Pisa, Italy [email protected] flavone, that at a concentration of 40 μg/mL was able to inhibit the biofilm formation of 50% ofS. Mentha piperita is a prototypical member of the aureus and of 90% of C. albicans. mint family and has wide adaptation ability in dif- 1. Midiwo, J.O.; Owuor, F.A.O.; Juma, B.F.; Waterman, P.G. Diter- ferent climate and soil conditions, but temperate penes from the leaf exudate of Psiadia punctulata Phytochem- climates are more suitable for its high quality [1]. istry. 1997;45(1):117-20. The yields and phytochemicals of Mentha as in other 2. Xiaoping, Z.; Bremer, K. A cladistic analysis of the tribe Astereae (Asteraceae) with notes on their evolution and subtribal classifi- medicinal plants are influenced by various factors, cation Plant Syst Evol. 1993;184:259-83. such as geographical location [2], environmental 3. Greuter, W.; The Euro Med treatment of Astereae (Composi- conditions [3] and agro-climatic requirements of the tae) – generic concepts and required new names. Willdenowia. crops [4]. Agro-climatic zone is a land unit in terms 2003;33:45-7. 4. Juma, B.F.; Midiwo, J.O.; Yenesew. A. Waterman, P.G.; Heyd- of major climate, superimposed on length of grow- enreich, M. Peter, M.G. Three ent-trachylobane diterpenes ing period i.e. moisture availability period [5]. The from the leaf exudates of Psiadia punctulata. Phytochemistry. Egypt land can be classified to eight main agro-eco- 2006;67:1322–5. 5. Juma, B.F.; Yenesew, A.; Midiwo, J.O.; Waterman, P.G.; Fla- logical zones according to evapotranspiration (ETo) vones and phenylpropenoids in the surface exudate of Psiadia [6].This investigation was carried out during two punctulata. Phytochemistry. 2001;57:571–4. successive seasons to evaluate the growth produc- 6. Abou-Zaid, M.M.; El-Karemy, Z.; El-Negoumy, A.I.; Altosaar, tivity and oil quality of Mentha piperita var. Multy- I.; Saleh, N.A.M.; The flavonoids ofPsiadia punctulata. Bull Chem Soc Ethiop. 1991;5(1):37-40. mentha grown in four different locations in Egypt

giugno 2017 natural 1 t 65 ABSTRACT

(El-Sharkia, El-Fayoum, Ismailia and North Sinai). Effect of Ribes nigrum bud The obtained results refer to presence of high signif- extract on human gingival icant differences between locations under study and mesenchymal stem cell a positive correlation could be observed between cytokine release: a role on the yield of Mentha plants and temperate climate regenerative processes conditions. The samples from El-Fayoum location showed the highest record for all growth parameters Chiara Giacomelli1, Letizia Natali1, Marinella than other locations. On the other hand, there is a De Leo 1, Simona Daniele1, Marco Nisi2, Filippo great variability in the yield of essential oils where Graziani2,3, Alessandra Braca1,3, M. Letizia the EO ranged from 0.285 to 1.240% according to Trincavelli1,3, Claudia Martini1,3 the agro-ecological region of cultivation. The chemical composition of EOs obtained from different loca- 1 Department of Pharmacy, University of Pisa, Via Bonanno 6, tions are characterized by high menthone/menthol 56126 Pisa, Italy; 2 contents and high antioxidant activity especially in Department of Surgical, Medical, Molecular and Critical Area Pathology, University of Pisa, Via Savi 10, 56126 Pisa, Italy; the plants grown in El-Fayoum and Ismailia which 3 Centro Interdipartimentale di Ricerca “Nutraceutica e could be attributed to their high amount of pheno- Alimentazione per la Salute”, University of Pisa, via del Borghetto lic compounds and flavonoids comparing to other 80, 56124 Pisa, Italy Department of Pharmacy, University of Pisa, Via Bonanno 6, locations. The amount of oxygenated compounds in 56126 Pisa, Italy the EOs is high in the warmer locations (El-Fayoum [email protected] 89.62%), while the amount of sesquiterpene hydro- carbons was found to be high in El-Sharkia locations The blackcurrant, Ribes nigrum, has been widely (6.68%). Cultivation of Mentha piperita var. Multy- used in the traditional herbal medicine, and recently mentha in different Egyptian locations characterized its anti-inflammatory, antioxidant and regenerative by different geographical and weather conditions, af- properties have been highlighted1. In the scenario fected the quantities of the major components in a of regenerative medicine, the gingival mesenchymal statistically significant way. stem cells (GMSCs) have arisen as a promising tool to repair damaged tissues. The GMSCs are a pecu- 1 Singh, R.; Muftah, A.M. ; Belkheir, A. Antibacterial and antioxi- liar type of mesenchymal stem cell that besides the dant activities of Mentha piperita L. Arabian Journal of Chemis- try, (2015): 8: 322–328. immunomodulatory and differentiation properties 2 RajeswaraRao, B.R. Biomass and essential oil yields of corn mint they behave as trophic mediators2; furthermore, (Mentha arvensis L. f. piperascens Malinvaud ex Holmes) plant- they could be easily obtained from gingival lamina ed in different months in semi-arid tropical climate. Ind. Crops propria3. Herein, the GMSCs were isolated and char- Prod. 1990: 10, 107-113. 3 Chalchat, J.C.; Garry, R.P.; Michet, A. Variation of the chemical acterized from healthy subjects, and a cell pool was composition of essential oil of Mentha piperita L. during the used to investigate the regenerative effect of a R. ni- growing time. J. Essent. Res. 1997: 9, 463-465. grum bud extract. 4 Chand, S.; Patra, N.K.; Anwar, M.; Patra, D.D. Agronomy and uses of menthol mint (Mentha arvensis) Indian prospective. The R. nigrum bud extract was able to increase Proc. Indian Nat. Sci. Acad. 2004: B70 (3), 269-297. the GMSC proliferation and the gene expression 5 FAO, Land Evaluation of Rain-fed Agriculture. Soil Bull. 1983: of stemness markers (SOX2, Oct4) at least at high 52-FAO, Rome, 237p (1983). concentrations. Classic growth factors and cyto- 6 Khalil F., Ouda S., Osman N.A., and Ghamis A. Determination of Agro-Climatic Zones in Egypt Using a Robust Statistical Pro- kines, such as TGF-β, interleukins (IL-6, IL-10), and cedure. Fifteenth International Water Technology Conference, COX2 serve as paracrine control molecules secreted IWTC-15 2011, Alexandria, Egypt or packaged into extracellular vesicles by GMSCs, in response to an inflammatory stimuli (100 ng/ml TNF-α). The extract (50 µg/ml) was able to counteract the secretion of these inflammatory mediators. The presence of high TNF-α concentration affected the secretome of the GMSCs, inducing a decrease of the endothelial cells migration, so impairing the re- pairing processes. Interestingly, the negative effect of TNF-α was counteract by the R. nigrum bud extract that significantly restore the migration ability of the endothelial cells. Furthermore, the bud extract per se was able to significantly enhance the proliferation of the endothelial cells. Results from the HPLC-PDA/UV-ESI-MS/MS analy- Ribes nigrum

66 t natural 1 giugno 2017 ses revealed that R. nigrum bud preparation is rich 1 Department of Pharmaceutical Sciences (DISFARM), in polyphenol constituents, according to data pre- University of Milan, Via Mangiagalli 25, I-20133 Milan, Italy; ; 2 Mass Spectrometry Center (CISM), University of Florence, Via 4 viously reported. In particular, phenolic acids de- U. Schiff 6, I-50019 Sesto F.no, Florence, Italy; rivatives and flavonol mono- and di-glycosides were 3 Department of Biology (BIO), University of Florence, Via G. La identified. In conclusion, these results highlight the Pira 4, I-50121 Florence, Italy; 4 Department of Plant, Soil and Environmental Sciences R. nigrum bud extract as a novel tool to enhance (DISPAA), University of Florence, Piazzale delle Cascine 28, the reparative processes, through its direct effect I-50144 Florence, Italy on endothelial cells and through the modulation of [email protected] cytokine release from GMSCs. Furthermore, the results could open the way to the development of the Ceratozamia (Zamiaceae) is a genus of New extract as topic agent in wound healing as well as in World Cycads, containing more than 20 species periodontal regeneration. distributed in eastern and southern Mexico, with extensions into Central America1. Morphology, 1. Gopalan, A.; Reuben, S.C.; Ahmed, S.; Darvesh, A.S.; Hohmann, anatomy, and molecular data allowed resolution of J.; Bishayee, A. The health benefits of blackcurrants. Food Funct. 2012, 3, 795-809. some aspects of species relationships, suggesting 2. Hofer, H.R.; Tuan, R.S. Secreted trophic factors of mesenchymal phytogeographic hypotheses2. Renewed interest stem cells support neurovascular and musculoskeletal thera- in cycads has also resulted in works focused on pies. Stem Cell Res. Ther. 2016, 7, 131. phytochemical topics3, however data on the volatile 3. Fawzy El-Sayed, K.M.; Dörfer, C.E. Gingival Mesenchymal Stem/Progenitor Cells: A Unique Tissue Engineering Gem. Stem emission profiles lack. Cells Int. 2016, 2016, 7154327. Seven Ceratozamia species belonging to clade 4. Donno, D.; Beccaro, G.L.; Mellano, M.G.; Cerutti, A.K.; Bounous “C” 2 and cultivated in a cold greenhouse at the G. Medicinal plants, chemical composition and quality: may blackcurrant buds and blackberry sprouts be a new polyphenol Florence Botanic Garden were investigated: C. source for herbal preparations? J Appl Bot Food Qual 2013, 86, fuscoviridis D. Moore, C. hildae Landry & M. 79-89. Wilson, C. kuesteriana Regel, C. latifolia Miq., C. longifolia Miq., C. mexicana Brongn., C. vulgaris J. Schust. Perennial leaves were screened in February, July and September 2011 for the emission of volatile organic compounds (VOCs) by means of headspace solid phase microextraction coupled with gaschromatography and mass spectrometry (HS-SPME-GC/MS); two leaflets per species were sampled from coetaneous leaves, and analysed. In this study we assessed and compared the volatile profiles among species and across sampling periods, with the main objective to collect preliminary information to evaluate the potentiality of VOCs for taxonomic purposes. The VOC analysis revealed the emission of overall 199 compounds, of which 144 identified. The unknown compounds dominated the volatile profiles, accounting for about 27%, followed by aldehydes, terpenes, alcohols, ketones and esters. The highest number of compounds (115) was found in C. fuscoviridis, the lowest (91) in C. longifolia. In all the examined species, February profiles

Foto di Krzysztof Ziarnek, Kenraiz Foto di Krzysztof Ziarnek, invariably exhibited the major amount of volatiles in Ceratozamia mexicana comparison to July and September. Statistical data processing proved: (i) a low level of diversity among the VOC profiles of the examined Volatile profile of perennial le- species referred to each collection time; (ii) a great aves of seven cultivated species variability in the temporal emission patterns of all of Ceratozamia (cycads) the species across the three sampling periods. These evidences revealed the predominance of Claudia Giuliani1, Riccardo Romoli2, Roberto the environmental components at the study area Calamassi3, Luca Calamai4, Gelsomina Fico1 in affecting the volatile profiles ofCeratozamia

giugno 2017 natural 1 t 67 ABSTRACT

species, at the expense of the metabolic resilience in the pharmacological potential of natural prod- related to the different climatic conditions of the ucts [3]. Salvia are being used extensively in folk native regions. Possible ecological roles of the medicine in Jordan and many other Mediterranean VOC temporal variation in relation to the plant countries and this study aimed to investigate the phenological stages are discussed. ability of S. fruticosa to inhibit pro-inflammatory cytokines production[4]. We first examined the in- 1 Vovides A.P.; González, D.; Pérez-Farrera, M.A.; Avendaño, S.; hibitory effects of Greek or Mediterranean sage S. Bárcenas, C. A review of research on the cycad genus Ceratozamia Brongn.(Zamiaceae) in Mexico. Taxon 2004, 53, 291-297. fruticosa methanolic crude extract on TNF-α, IL-1β 2 Gonzáles, D.; Vovides, A.P. (). Low intralineage divergence in and IL-6 production in different murine cell types Ceratozamia (Zamiaceae) detected with nuclear ribosomal DNA as in vitro cellular models and in LPS-challenged ITS and chloroplast DNA trnL-F non-coding region. Syst Bot., Balb/c mice as an in vivo animal model. 2002, 27, 654-661 3 Cardini, F.; Morassi Bonzi, L. Carotenoid composition and its The results obtained showed a potent inhibitory chemotaxonomic significance in leaves of ten species of the genus effect of S. fruticosa on the production level of Ceratozamia (Cycads). J Plant Physiol, 2005, 52, 517-528 pro-inflammatory cytokines in both cellular and animal models. Adjuvant-induced arthritis in rats was used as a recognized model represents a systemic inflammatory disease with bone and cartilage changes similar to those observed in humans rheumatoid arthritis[5],[6]. Rats treated with S. fruticosa showed no clinical manifestations, no significant increase in paw volume and no significant radiological observa- tions related to development of arthritis. Moreover, there was no significant difference between treated group and normal control group. Whereas, the arthritic group showed all clinical manifestations, increased paw volume and typical radiological findings of subchondral sclerosis, decreased joint space and soft tissue swelling which are the hallmarks of RA, in addition to behavioral changes related to the Salvia fruticosa painful nature of the disease. Immunomodulatory potential of It seems likely that S. fruticosa may act at various the Mediterranean sage therapeutic targets in RA and it may offer a promising anti-inflammatory and anti-rheumatic agent for (Salvia fruticosa) drug discovery researches. More experimental and in adjuvant-induced clinical trials are also needed to validate the useful- arthritis in rats ness of its use either alone or in combination with existing therapy. Jameel Fayyad Bzour

Assistant Professor 1. Taylor, P.; Feldmann, M. Rheumatoid Arthritis: Pathogenic Zarqa University, Faculty of Allied Medical Sciences, Department Mechanisms and Therapeutic Targets. Drug Discov Today. of Medical Technology, P.O. Box 132222, Zarqa 13132 Jordan 2004, 3, 289-295. [email protected] 2. Firestein, S. Inhibiting Inflammation in Rheumatoid Arthritis. N Engl J Med. 2006, 354, 80-82. 3. Spelman, K.; Burns, J.; Nichols, D.; Winters, N.; Ottersberg, S.; Inflammatory arthritis, in particular rheumatoid Tenborg, M. Modulation of Cytokine Expression by Tradition- arthritis (RA), is a chronic inflammatory disease al Medicines: A Review of Herbal Immunomodulators, Altern which is an important health problem for millions Med Rev. 2006, 11(2), 128-150. 4. Singh, D.; Aggarwal, A.; Mathias, A.; Naik, S. Immunomodu- of people worldwide. It has been the focus of in- latory Activity of Semecarpus anacardium Extract in Mono- tense investigation, but its etiology and pathogene- nuclear Cells of Normal Individuals and Rheumatoid Arthritis sis remains controversial [1]. Patients. J Ethnopharmacol. 2006 108(3), 398-406. 5. Barsante, M.; Roffê, E.; Yokoro, C.; Tafuri, W.; Souza, D.; Pin- The current pharmacological treatment continues ho, V.; Castro, M.; Teixeira, M. Anti-inflammatory and Analge- to be inadequate in preventing the progression of sic Effects of Atorvastatin in ARat Model of Adjuvant-Induced this disease to the stage of irreversible joint ero- Arthritis. Eur J Pharmacol. 2005, 516, 282-289. sion and deformity [2]. Therefore, development of 6. Jawed, H.; Shah, S.;. Jamall, S.; Simjee, S. N-(2-hydroxy phenyl) Acetamide Inhibits Inflammation-Related Cytokines and new anti-inflammatory and anti-rheumatic drugs ROS in Adjuvant-Induced Arthritic (AIA) Rats. Int Immuno- continues to be essential. There is growing interest pharmacol. 2010, 10, 900-905.

68 t natural 1 giugno 2017 or subcutaneous administration of GRA or SFN as well as TBK-sprouts extract (equimolar dosage of GRA) reduced the hypersensitivity to cold non-noxious stimuli (allodynia-related measurement) in a dose-dependent

manner. The H2S-binding molecule Hb abolished the pain-relieving effects of GRA and SFN. The anti-neuropathic properties of both molecules were reverted by the Kv7 potassium channel blocker XE991. More- over, to evaluate possible preventive effects, GRA and SFN were administered in a different group once daily p.o. and s.c. starting from the first day of oxaliplatin injection until the 14th and behavioural test were performed on days 7 and 14. Repeated GRA and SFN administration completely prevented the chemotherapy-induced hypersensitivity to thermal stimuli back to the level of healthy controls. GRA and SFN reduce

neuropathic pain by releasing H2S and show a protec- Cavolo nero tive effect against the development of chemotherapy-induced neuropathy. The anti-hyperalgesic effect of TBK-sprouts extract is closely related to the content Tuscan black kale as source of GRA. SFN is three times more potent than GRA of compounds active on suggesting that the glucosinolate must be metabolized chemotherapy-induced to isothiocyanate before releasing H2S. The anti-hy- neuropathic pain: a hypothesis peralgesic effect is largely mediated by the activation about the mechanism of action of Kv7 channels.

Elena Lucarinia, Lorenzo Di Cesare Mannellia, Lau- 1. De Nicola, G.R.; Rollin, P.; Mazzon, E.; Iori, R. Novel gram-scale a a a production of enantiopure R-sulforaphane from Tuscan black ra Micheli , Mario Maresca , Barbara Tenci , Renato kale seeds. Molecules. 2014, 19(6), 6975-6986. b c a Iori , Vincenzo Calderone , Carla Ghelardini 2. Citi, V.; Martelli, A.; Testai, L.; Marino, A.; Breschi, M.C.; Cal- derone, V. Hydrogen sulfide releasing capacity of natural isothio- aDept. of Neurofarba, University of Florence, Viale Pieraccini 6, cyanates: is it a reliable explanation for the multiple biological 50139, Florence, Italy. bCREA-CIN, Via di Corticella 133, 40128, effects of Brassicaceae? Planta Med. 2014, 80(8-9), 610-613. Bologna, Italy. cDept. of Pharmacy, University of Pisa, Via Bonan- 3. Martelli, A.; Testai, L.; Citi, V.; Marino, A.; Bellagambi, F.G.; no 6, 56126, Pisa, Italy. Ghimenti, S.; Breschi, M.C.; Calderone, V. Pharmacological [email protected] characterization of the vascular effects of aryl isothiocyanates: is hydrogen sulfide the real player? Vascul. Pharmacol. 2014, l60(1), 32-41. The glucosinolate glucoraphanin (GRA), found nota- 4. Busserolles, J.; Tsantoulas, C.; Eschalier, A.; García, J.A.L. Po- bly in Tuscan black kale (TBK, Brassica oleracea L. tassium channels in neuropathic pain: advances, challenges, and var. acephala sabellica), may release by the enzy- emerging ideas. Pain. 2016, 157 Suppl 1, S7-14. matic action of myrosinase the isothiocyanate sulforaphane (SFN)1. Recently, the release of hydrogen sul- phide (H2S) has been related to many pharmacological properties of isothiocyanates-based compounds2. The gasotransmitter can modulates the activity of Kv73, a class of voltage-gated potassium channels which plays a pivotal role in pain modulation4. The aim of the present study was to evaluate the effectiveness of a TBK-sprouts extract enriched in GRA in reducing

neuropathic pain induced by chemotherapeutic drugs. Agostinelli Foto di B. Pain relieving effects of GRA and SFN were studied to individuate their role in the pharmacodynamic of the extract. With this purpose the importance of H2S releasing ability and the potential involvement of Kv7 potassium channels in pain modulation of these compounds were investigated. Mice were treated daily with oxaliplatin to induce neuropathic pain and acute behavioral tests were performed on day 15. A single oral Hypericum androsaemum

giugno 2017 natural 1 t 69 ABSTRACT

Evaluation of Hypericum ined by measuring the TBARS, SOD, Cat and GSH androsaemum L. red fruit levels. effects in a mouse model of Finally, the content of ten bioactive compounds in post-stroke depression the H. androsaemum red berry extracts was mea- sured by HPLC-DAD analysis. Seyed Mohammad Nabavia, Seyed Fazel Nabavia, Results showed that H. androsaemum red berry Antoni Suredab, Giovanni Capriolic, Romilde aqueous extracts reduced the overall symptoms of Iannarellic, Arold Jorel Tsetegho Sokengd, Maria post-stroke depression, restoring near-normal be- Dagliad, Nady Braidye, Sedigheh Khanjanif, Akbar havior. Correlation between pharmacological re- Hajizadeh Moghaddamg, Filippo Maggic sults and the presence in the fruit extract of some bioactive compounds was discussed. a Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran 1 Lloyd-Jones, D.; Adams, R.; Carnethon, M.; De Simone, G.; Fer- b Research Group on Community Nutrition and Oxidative Stress, guson, T.B.; Flegal, K.; Ford, E.; Furie, K.; Go, A.; Greenlund, Lab of Physical Activity Sciences, IUNICS, University of Balearic K. Heart disease and stroke statistics—2009 update a report Islands, Palma de Mallorca, Spain from the American Heart Association Statistics Committee c School of Pharmacy, University of Camerino, Camerino, Italy and Stroke Statistics Subcommittee. Circulation 2009, 119, d Department of Drug Sciences, Medicinal Chemistry and Phar- e21-e181. 2 Nabavi, S.-F.; Dean, O.-M.; Turner, A.; Sureda, A., maceutical Technology Section, University of Pavia, Pavia Italy Daglia, M.; Nabavi, S.-M. Oxidative stress and post-stroke de- e Centre for Healthy Brain Ageing, School of Psychiatry, Universi- pression: possible therapeutic role of polyphenols? Curr. Med. ty of New South Wales, Australia Chem. 2015, 22, 343–351. 3 Caprioli, G.; Alunno, A.; Beghelli, f Department of Physiology, Faculty of Biological Sciences, Shahid D.; Bianco, A.; Bramucci, M.; Frezza, C.; Iannarelli, R.; Papa, Behshti University, Tehran, Iran F.; Quassinti, L.; Sagratini, G.; Tirillini, B.; Venditti, A.; Vit- g Department of Biology, Faculty of Basic Sciences, University of tori, S.; Maggi, F. Polar constituents and biological activity of Mazandaran, Iran the berry-like fruits of Hypericum androsaemum. Front. Plant [email protected] Sci. 2016, 7, 232. 4 Antognoni, F.; Lianza, M.; Poli, F.; Buc- cioni, M.; Santinelli, C.; Caprioli, G.; Iannarelli, R.; Lupidi, G.; Stroke is one of the most important cerebrovascu- Damiani, E.; Beghelli, D.; Alunno, A.; Maggi, F. Polar extracts lar diseases, and is classified into two categories, from the berry-like fruits of Hypericum androsaemum L. as a hemorrhagic and ischemic stroke, with the latter promising ingredient in skin care formulations. J. Ethnophar- macol. 2017, 195, 255-265. causing infarction and brain injury during which alteration of antioxidant defenses and generation of reactive oxygen species (ROS) cause neuronal dysfunction and death.1 Oxidative stress thus plays a pivotal role for the development of major depres- sive disorder (MDD). Indeed, depression is an important and frequent neuropsychiatric disorder occurring in stroke patients.2 Searching for medicinal plants endowed with antidepressant and antioxidant properties, thus potentially useful in the treatment of post- stroke depression, we focused our attention on Hypericum androsaemum L., an evergreen shrub growing in thermophilous woods and considered one of the most used traditional remedies in the genus Hypericum throughout the Mediterranean area.3,4 In particular, we assumed that the antioxidant potential of H. androsaemum berry-like fruits, previously demonstrated,3,4 may be helpful Echinophora tenuifolia in treating the symptoms of post-stroke depression. For the purpose, a mouse model of post-stroke depression was used. Echinophora tenuifolia L. Ischemic stroke was induced in mice by bilateral aerial parts: phytochemical common carotid artery occlusion and the antide- constituents and pressive-like effects of two aqueous extracts of H. antiproliferative activity androsaemum red fruits at two doses (15 and 30 against human cancer cell lines mg/kg bw i.p.) were evaluated by despair swimming test (DST) and tail suspension test (TST). Further- Mariangela Marrelli, Giancarlo Statti, more, the oxidative stress in mice brain was exam- Filomena Conforti

70 t natural 1 giugno 2017 Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, I- 87036 Rende (CS), Italy, [email protected]

Echinophora tenuifolia L. belongs to the Apiaceae family and is one of the ten species of this genus distributed in Mediterranean and Middle East re- gions1. In Turkish traditional medicine this plant was used as a remedy against wounds and gastric Longo Foto di R. ulcers due to its antifungal, carminative and digestive properties2. Some Echinophora species are usually added to soup, meat, cheese and yoghourt to enhance their sensory properties and, recently, the essential oil of E. platyloba DC. has been used as a natural pre- servative in dairy products industries to prevent fungal contaminations3,4. A number of studies dealt with the phytochemical composition of essential oils of different species, above all E. platyloba DC. and E. sibthorpiana (Guss.) Tutin, but till now, to the best of our knowledge, a few number of works focused on the biological activity of E. tenuifolia L.

The aim of this study was to evaluate the antipro- Crocus sativus liferative activity against human cancer cell lines of different aerial parts of E. tenuifolia L.: branches and inflorescences. Samples were extracted Volatile organic compounds and through maceration and the resultant raw extracts antioxidant activity of were fractionated using solvents with increasing non-polar extract from Saffron polarity: n-hexane, dichloromethane and ethyl ace- stamens harvested in Morocco tate. Total phenolic and flavonoid contents of both branches and inflorescences were evaluated, and G.L. Petrettoa, K. Bakhyb, M.Lageb, C. Al Faizb, polar and non polar constituents were identified as F. Fancelloc, S. Zarac, R. Addisa, well. M. Chessaa and G. Pintorea The antioxidant properties of E. tenuifolia L. sam- aDepartment of Chemistry and Pharmacy, ples were also investigated by means of DPPH and University of Sassari, Italy β-carotene bleaching test. bInstitut national de la Recherche Agronomique (INRA), BP 6570 The in vitro antiproliferative activity of alcoholic Rabat-Instituts, Maroc cDipartimento di Agraria, University of Sassari, italy extracts and their fractions was assessed against hu- [email protected] man cancer cell lines by MTT assay. The dichloromethane fraction of branches extract, particularly, Crocus sativus Linn. (Iridaceae), commonly known caused a significant inhibition of cell viability. as saffron is the raw material for the production of the world’s most costly spice which is famous for its 1 Georgiou, C.; Koutsaviti, A.; Bazos, I. ; Tzakou O. Chemical distinct color, taste, aroma and medicinal properties. composition of Echinophora tenuifolia subsp. sibthorpiana Among the whole saffron flower only the stigmas are essential oil from Greece. Rec. Nat. Prod. 2010, 4, 167-170. 2 Glamoclija, J.M.; Sokovic, M.D.; Šiljegovic, J.D.; Ristic, collected and processed for spice production. The M.S.; Ciric, A.D.; Grubišic D.V. Chemical composition and stigmas represent a very small percent of the total antimicrobial activity of Echinophora spinosa L. (Apiaceae) flower and the production of saffron spice generate a essential oil. Rec. Nat. Prod. 2011, 5, 319-323. great amount of by-products. 3 Asghari, J.; Touli, C. K.; Mazaheritehrani M. Microwave-assisted Despite the great number of papers focused on the hydrodistillation of essential oils from Echinophora platyloba characterization of stigmas1, very little is known on DC. J. Med. Plants Res. 2012, 6, 4475-4480. 4 Gokbulut, I.; Bilenler, T.; Karabulut I. Determination of the other saffron organs. chemical composition, total phenolic, antimicrobial, and The colour of saffron stigmas (saffron name derived antioxidant activities of Echinophora tenuifolia essential oil. from the translation of the Arab word “yellow”) is Int. J. Food Prop. 2013, 16, 1442–1451. strictly related to the presence of carotenoid com-

giugno 2017 natural 1 t 71 ABSTRACT

pounds which are also the main responsible for the 1 University of Florence, Department of Chemistry, Via Ugo Schiff characteristic aroma. Enzymatic, photo-oxygen- 6, 50019 Sesto Fiorentino (FI), [email protected]; 2 University of Florence, Department of Neuroscience, Psycholo- ation, auto-oxidation and thermal degradation of ca- gy, Drug Research and Child Health, Section of Pharmacology and rotenoid compounds yield the aroma components2. Toxicology, Viale Gaetano Pieraccini 6, 50139 Firenze Since in the literature3 has been demonstrated the [email protected] presences of several carotenoids compounds also in The study is focused on the preparation and evalua- the discarded saffron stamens, the aim of this study tion of naked and chitosan-coated stealth solid lipid is then to characterize Moroccan stamen saffron non nanoparticles (SLNs) containing silibinin (Sb) to in- polar extract. Chemical characterization was carried crease its solubility and favour its intestinal absorption. out by GC and GC/MS analysis on the diethyl ether Sb is the key biologically active flavonolignan pres- extract as well as by solid phase micro extraction ent in silymarin isolated from the fruits and seed of (SPME) followed by GC analysis. Furthermore the Silybum marianum L. Gaertn.. Sb is used for treat- antioxidant and antimicrobial activity of the ether ment of acute and chronic liver toxicities, inflam- extract were evaluated. As comparison and control mation, fibrosis and oxidative stress. It is also active also the antioxidant activity and the total polyphe- against different carcinomas. Recently Sb has been nol content of methanolic extract were assessed. proposed to be beneficial in type 2 diabetes patients. 1. Lage, M.; Melai, B.; Cioni, P. L.; Flamini, G.; Gaboun, F.; Bakhy, However, its hydrophobicity nature limits bioavail- K.; Zouahri, A.; Pistelli, L. Phytochemical composition of Mo- ability compromising in vivo biological effects [1]. roccan saffron accessions by headspace solid-phase-microex- SLNs were designed as exceptionally safe colloidal traction. Am. J. Ess. Oils Nat. Prod. 2015, 2, 1-7. 2. Winterhalter; Rouseff;. 2002. Carotenoid derived aroma com- carriers for the oral delivery of poorly soluble drug. pounds: an introduction. ACS symposium series; American In this work SLNs were prepared by emulsion/evap- Chemical Society: Washington, DC, 2001. oration/solidifying method, using biodegradable and 3. Montoro, P.; Maldini, M.; Luciani, L.; Tuberoso, C.I.; Congiu, bioacceptable components such as stearic acid and F.; Pizza, C. Radical Scavenging Activity and LC-MS Metabolic Profiling of Petals, Stamens, and Flowers of Crocus sativus L. J. brij 78. In order to increase the absorption rate and Food Sci. 2012, 77, C893–900. improve the delivery efficacy of SLNs we applied a chitosan coating. Chitosan is a naturally occurring cationic poly- saccharide derived by deacetylation of chitin. The primary amino group offers some special properties such as water solubility and hemocompatibil- ity. Based on cationic property chitosan-coated nanoparticles exhibit mucoadhesive features and a Foto di F. Mearelli Foto di F. prolonged residence time in the negatively charged epithelia in small intestine, thus increasing the drug concentration at the site of absorption [2]. Developed SLNs exhibited particle size and z-potential in the range of 150-200 nm and -48 - +40 mV respectively. Encapsulation efficiency and recovery were greater than 90%. Moreover SLNs showed excellent stability in both simulated gastric fluid and simulated intestinal fluid with a small release of Sb during at least 24 hours. Further we demonstrated that SLNs were successful in increasing the permeation of Sb by Parallel Sylibum marianum Artificial Membrane Permeability Assay (PAMPA). Fluorescein isothiocyanate was selected as a model Solid lipid nanoparticles for dye to analyse in vitro uptake and permeation of oral delivery of silibinin: drug using Caco-2 cell line. The obtained results formulation, characterization suggest that the developed SLNs could improve the and in vitro evaluation using bioavailability of Sb. pampa and caco-2 cell models [1] Pepping, J. Milk thistle: Silybum marianum. American journal of health-system pharmacy. 1999, 56(12), 1195-1197. 1 1 1 Vieri Piazzini , Giulia Graverini , Giulia Vanti , [2] Fonte, P.; Andrade, F.; Araujo, F.; Andrade, C.; Neves, J. D.; Elisabetta Bigagli2, Lorenzo Cinci2, Cristina Luceri2, Sarmento, B. Chitosan-coated solid lipid nanoparticles for Anna Rita Bilia1, Maria Camilla Bergonzi1 insulin delivery. Methods Enzymol. 2012, 508, 295-314.

72 t natural 1 giugno 2017 gate the ex vivo antioxidant effect and the in vivo protective effect against hepatic steatosis in rats. We characterized the total polyphenol content (TPC), total flavonoid content (TFC), flavonols and anthocyanins of aqueous extract of Prunus spinosa. The polyphenolic profile was analyzed by HPLC- DAD. In human red blood cells we studied the antioxidant activity, by the Cellular Antioxidant Activity (CAA-RBC) test, and the capacity to inhibit hemolysis. Moreover, we treated rats with two doses of blackthorn (400 mg/kg b.w. and 800 mg/kg b.w.) for 28 days, after induction of hepatic steatosis by high fat diet and Sreptozotocin (45 mg/kg i.p.). The aqueous extract of blackthorn showed a good content of TPC (4.16±0.14 mg GAE/g), TFC (8.47±0.33 mg QE/g), flavonols (0.29±0.02 mg CE/g) and anthocyanins (0.06±0.03 mg Cyanidin EQ/g). The polyphenolic profile showed the presence of neochlorogenic acid, protocatechuic acid, chlorogenic acid, syringic acid, rutin, vitexin, elaigic acid, cynaroside, trans sinapic acid and trans ferulic acid. The CAA-RBC and percentage of hemolysis inhibition capacity were similar to the Trolox (standard). Rats treated with 800 mg/kg b.w. of Prunus spinosa showed a reduced hepatic lipid content compared to positive control group (162.2±35.5 vs 209.9±11.9 mg/g, p<0.05). In conclusion, the results Prunus spinosa of the present study highlighted the good content of bioactive compounds of Prunus spinosa, its ex vivo Chemical characterization of antioxidant activity and its protective effect against blackthron (Prunus spinosa) hepatic steatosis in rats.

fruit and evaluation of its 1. Calvo, M. I.; Akerreta, S.; Cavero, R. Y. Eur. J. Int. Med. 2013, protective effect against hepatic 5, 537–546. steatosis in rats 2. Veličković, J.M.; Kostić, D.A.; Stojanović, G.S.; Mitić, S.S.; Mitić, M.N.; Ranđelović, S.S.; Đorđević, A.S. Hem. Ind. 2014, 68 (3), 297–303. Luisa Pozzo1, Francesco Vizzarri2, Clara M. Della Croce1, Rossella Russo1, Marisa Palazzo2, Donato Casamassima2, Souid Aymen1,3, Martina Menchini1, Vincenzo Longo1

1Istituto di Biologia e Biotecnologia Agraria, CNR, Via Moruzzi 1, 56124, Pisa; 2Dipartimento di Ambiente, Agricoltura e Alimenti, Università del Molise, Via De Sanctis snc, 86100, Campobasso; 3Laboratoire des Plantes Extrêmophiles. Centre de Biotechnologie de Borj Cedria. BP 901, Hammam Lif 2050. Tunisia [email protected]

Blackthorn (Prunus spinosa L.), which belongs to the rose family (Rosaceae), is a perennial plant growing in moderate continental climate in northern hemisphere and off spread in the Southern regions of Italy, where the fruits are used to produce a liqueur. It is used in phytotherapy for the treatment of many diseases, due to its diuretic, spasmolytic, anti-septic and anti-inflammatory activity on the mucosal layer of the digestive system.1,2 The aim of this study was to characterize the blackthorn fruit and to investi- Daucus muricatus

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Phytochemical investigation on the Northern African endemic plant Daucus virgatus

Daniela Rigano1; Alì Snene2; Carmina Sirignano1; Carmen Formisano1; Sofia Tapanelli3, Angela Ianaro1; Saoussen Hammami2; Annette Habluetzel3, Orazio Taglialatela-Scafati1

1Department of Pharmacy, University Federico II, Via D. Foto di Joan Simon Montesano, 49, I-80131 Naples, Italy; 2Research Unit Applied Chemistry and Environment 13ES63, Monastir University, Faculty of Sciences of Monastir, 5000 Monastir, TUNISIA 3School of Pharmacy, University of Camerino, Piazza dei Costanti, 62032 Camerino, MC, Italy [email protected]

The genus Daucus (family Apiaceae) includes more than 80 accepted species distributed mostly in Europe, North Africa and West Asia; about five Daucus species are widespread in Tunisia. Daucus virgatus (Poir.) Maire grows in Northern Africa as an herbaceous annual or biennial species and is used in folk medicine in Tunisia1. The first phytochemical screening on D. virgatus aerial parts, collected Artemisia umbelliformis during the flowering stage, yielded to the isolation of a series of sesquiterpenoids in the pure form Artemisia umbelliformis Lam. belonging to distinct structural classes. Along with and génépi liqueur. five known metabolites, several new molecules From the field to the final product: the importance of the volatile fraction in quality control

Barbara Sgorbini, Lorenzo Boggia, Maria Laura Colombo, Arianna Marengo, Stefano Acquadro, Carlo Bicchi, Patrizia Rubiolo

Dipartimento di Scienza e Tecnologia del Farmaco (DSTF), Figure 1. Representative examples of isolated sesquiterpenoids University of Turin, Via Pietro Giuria 9, 10125 Torino, Italy - E-mail: barbara. (Figure 1) were identified and their stereostructures [email protected] determined by a detailed MS and NMR analysis. The isolated metabolites belonging to elemane-, Volatile fraction plays a fundamental role for quali- eudesmane- and daucane- type were evaluated and quantitative characterization of aromatic plants for their antiproliferative activity against three and their relative products, i.e. alcoholic beverages. human cell lines, A375 (melanoma), MCF-7 (breast A. umbelliformis, commonly known as “white adenocarcinoma), and HACAT (keratinocyte), while génépi”, is traditionally used to prepare a precious germacranes were assayed for their transmission liqueur, and it is characterized by a volatile fraction blocking effects on Plasmodium berghei early rich in two monoterpenoids, i.e. a- and β- thujone. sporogonic development. The structure elucidation The maximum content of thujones in Artemisia- and the results of biological evaluation of new based beverages is limited to 35 mg/L in the EU1 compounds will be presented. because of their recognized activity on human central nervous system. 1. Hostettmann, K.; Marston, A.; Ndjoko, K.; Wolfender, J.L. The This study reports the results of an investigation to potential of African plants as a source of drugs. Curr. Org. define the geographical origin and thujone content of Chem. 2000, 4, 973-1010.

74 t natural 1 giugno 2017 individual plants of A. umbelliformis from different ronmental Sciences, University of Messina, 98168 Messina, Italy; geographical sites, cultivated experimentally at [email protected] 2 Foundation Prof. Antonio Imbesi, University of Messina, Messi- a single site, and to predict the thujones content na, Italy in the resulting liqueurs, through their volatile fraction. Headspace Solid Phase Microextraction Non-drug Cannabis sativa L. (Cannabaceae) va- (HS-SPME) coupled with Gas Chromatography- rieties, known to contain less than 0.2% of Δ9- Mass Spectrometry (GC-MS) or directly with tetrahydrocannabinol in the whole plant, can be mass spectrometry (HS-SPME-MS) were used in grown for their fiber, seed or oil. Oilseed ofC. sativa combination with chemometric descriptive and varieties is considered one of the few seed oils that predictive tools. HS-SPME-MS was applied to speed- contains about 80% polyunsaturated fatty acids in a up the total analysis time and to make the adopted perfect 3:1 ratio of Omega-6 to Omega-3, which is method suitable for the screening of a large number suggested as optimal for human nutrition1. Further- of samples. more, the antioxidant properties of cold-pressed seed With both approaches, a diagnostic prediction of: i) oil from cultivar of Cannabis sativa L.2, determines plant geographical origin, and ii) thujones content of its potential application in health promotion and dis- plant-related liqueurs could be achieved. ease prevention from oxidative damages. The antioxidants in edible oils include phenolic compounds that 1. Regulation (EC) No 110/2008 of the European Parliament and of the Council, Off. J. Eur. Union 2008, 16–54. act as anti-oxidants, or specific anti-oxidants such as the tocopherols. In our study, seed oil and its byprod- uct (sludge) from three Cannabis sativa L. varieties such as felina 32, uso 31 and futura 75 showed significant antioxidant properties in several methodologies in vitro. Polyphenolic compounds of the seed oils tested for antioxidant activity were characterized by HPLC-DAD. While, their fatty acids composition was analyzed by GC-MS. The results revealed that cold-pressed seed oils felina 32, uso 31 and futura 75, contain different classes of flavonoids such as flavonols, isoflavones, flavanones, flavones as well as significant amounts of phenolic acids, at different concentrations. The antioxidant activity was higher for the seed oils assayed than their byproducts and with the following order of potency felina 32 > uso 31 > futura 75. The different degree of antioxidant activity shown most likely may be due to different amounts of the phenolic compound classes contained in seed oils. The content of polyunsaturated fatty acids (PUFA) in seed oils tested was highlighted in the optimal 3:1 ratio of Omega-6 to Omega-3. Furthermore, stearidonic acid (SDA; 18:4, n-3), a long-chain PUFA (LCPUFA) that is a metabolic intermediate in the conversion of α-linolenic acid (ALA; 18:3, n-3) to eicosapentaeno- Cannabis sativa ic acid (EPA; 20:5, n-3) and docosahexaenoic acid (DHA; 22:6, n-3), it has been highlighted in significant Polyphenolic profile and fatty amount in all samples tested. These findings, suggest acids composition of seed oils that seed oils and their byproducts from felina 32, and their byproducts from uso 31 and futura 75 varieties of Cannabis sativa L., Cannabis sativa L. varieties with possess potential health-promoting properties. potential health-promoting properties 1. Scorletti, E.; Byrne, C.D. Omega-3 fatty acids, hepatic lipid me- tabolism, and nonalcoholic fatty liver disease. Annu. Rev. Nutr. 2013, 33, 231–248. Antonella Smeriglio1,2, Enza M. Galati1, Gianluigi 2. Smeriglio, A.; Galati, E.M.; Monforte, M.T.; Lanuzza, F.; D’An- Aliberto1,2, Maria T. Monforte1 and Clara Circosta1 gelo, V.; Circosta, C. Polyphenolic Compounds and Antioxidant Activity of Cold-Pressed Seed Oil from Finola Cultivar of Canna- 1Department of Chemical, Biological, Pharmaceutical and Envi- bis sativa L. Phytother. Res. 2016, 30, 1298–1307.

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vestigate how plant organisms respond to environmental factors (e.g. biotic and abiotic stress), aging or disease. In biotechnology, where plant cells are modified by the expression of engineered genes we can regulate the production of particular metabolites in plants to improve their adaptation to environmental stress, to improve food quality, and to increase crop yield. In our project, we use greenhouse and microplot cultivation of Belamcanda chinensis L. DC. (Iridaceae), Scutellaria baicalensis Georgi (Lamiaceae), Salvia miltiorrhiza L. (Lamiaceae), Perovskia atriplicifolia Benth. (Lamiaceae). We use aeroponic cultivation of the same plants to increase root biomass and increase of specific metabolites as well as for easy access to uncontaminated roots for precise metabolomic analyses. For data acquisition we use high resolution mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) , both methods are currently the most popular for metabolome identification.

1. Kim, HK.; Choi, YH.; Verpoorte, R. NMR-based plant metabolomics: where do we stand, where do we go? Trends Biotechnol. 2011, 29 , 6 , 267-275. Belamcanda chinensis 2. Rajalahti, T.; Kvalheim, OM. Multivariate data analysis in pharma- ceutics: A tutorial review. Int J Pharm. 2011,417, 1-2 , 280- 290. 3. Arbona, V.; Manzi, M.; Ollas, C.De.; Gómez-Cadenas, A. Practical aspects of using Metabolomics as a tool to investigate abiotic stress tolerance in metabolomic approach in plants. Int J Mol Sci. 2013, 14 , 3 , 4885-4911. medicinal plants analysis Acknowledgements: Study was supported by the Polish Nation- focusing on roots from al Science Center postdoctoral grant” to S. Ślusarczyk (No. aeroponics. FUGA - DEC/2014/12/S/NZ9/00715) .

Sylwester Ślusarczyk a,b Joanna Jaśpińskab Adam Matkowskib Mariusz Kowalczyka Anna Stochmala

a Department of Biochemistry and Crop Quality , IUNG-Institute of Soil Science and Plant Cultivation, State Research Institute, Czartoryskich 8, 24-100 Pulawy, Poland, b Department of Pharmaceutical Biology and Botany, Botanical Garden of Medicinal Plants, Wroclaw Medical University, Jana Kochanowskiego 10-14, Wroclaw, Poland [email protected]

Plants metabolomics is a quite new discipline growing rapidly for last 10 years and has received increasing attention in many areas of plant sciences 3.0) media - CC BY-SA Foto di (Lynk .1 Metabolomics means identification and quantifica- tion of all metabolites both primary and secondary in biological system under certain conditions. Plant metabolomics provided an overview of phytochemicals in cells, tissues, organs, and whole plant and give us more information and complete picture of living organisms. Many applications are based on metabolomic approach: drug discovery from natural resources, discovering lead compounds, quality control of herbal material, optimizing of agronomic 2,3 measures, or discrimination of taxa . We can in- Amaranthus retroflexus

76 t natural 1 giugno 2017 HPLC investigation of the Phytochemical analysis of nutraceutical composition of Cichorium intybus L. varieties the edible herb Amaranthus from Veneto (Italy) region retroflexus L. by means of HPLC-UV/DAD and HPLC-MS/MS Vito Alessandro Taddeo, Francesco Epifano, Serena Fiorito, Salvatore Genovese. Roberta Tardugnoa, Michele Pozzebonb, Marica Beg- giob, Giulio Pojanaa Dipartimento di Farmacia, Università “G. d’Annunzio” Chieti-Pescara, Via dei Vestini 31, 66100 Chieti Scalo (CH), Italy aDepartment of Philosophy and Cultural Heritage, University of [email protected] Venice, Dorsoduro 3484/D, 30123, Venice, Italy.

In this paper the presence of selected prenylated bDtoLabs, via Fratta, 25, 31023 Resana (TV), Italy. and unprenylated phenylpropanoids of nutraceutical value, namely umbelliferone, apigenin, 4’-ge- [email protected] ranyloxyferulic acid, 7-isopentenyloxycoumarin, au- The scientific interest in edible plants as potential raptene, and umbelliprenin have been determined in sources of health-beneficial bioactive compounds all parts of the edible herb Amaranthus retroflexus increased significantly in the last decades. The Med- extracted with different methodologies. Roots were iterranean basin and expecially Italy, are naturally seen to contain the widest variety of unprenylated rich in endemic plants consumed as traditional food and prenylated phenylpropanoids both in terms of with well known beneficial effects. Vegetables from number of secondary metabolites and their quanti- this territory are attracting great attention due to the tites. ABTS+• scavenging assay[1] revealed that its eth- large variety of compounds with bioactive properties anol extract exerted a powerful anti-oxidant activi- produced by various genera and species1. Among edi- ty. Findings described in the present study underline ble plants, Cichorium intybus L. (fam. Asteraceae), how A. retroflexuscan be considered as a valuable commonly known as ‘red chicory’ or ‘radicchio’, a nutraceutical having beneficial effects for human late winter salad cultivated in the Northern-East of welfare with great potentialities. Italy (Veneto region) can be a potential source for nutraceuticals since is known to contain bioactive natural antioxidants2. Red chicory, particularly resistant to low temperatures, is mostly consumed as raw salad or stewed in wintertime, when most vegetables are not available2, 3. This study aimed at investigating the antioxidant fingerprint of several C. intybus varieties, with particular interest in flavonoids and anthocyanins.

[1] Fan, R., Lin, N., Jiang, X.X., Yuan, F., Gao, Y.X. (2015). “HPLC- Various techniques were compared for the extraction DAD-MS/MS identification and ABTS+• on-line antioxidant activity of target compounds, including maceration and ul- evaluation of bioactive compounds in liquorice (Glycyrrhiza trasound-assisted extraction in order to obtain high uralensis Fisch.) extract”. European Food Research and Technology 240, 1035-1048. recoveries of polyphenols. The chemical analyses were carried out by Reversed phase -HPLC coupled with UV/DAD and jet stream ESI-MS/MS detection. A phenyl hexyl stationary phase was selected and applied under gradient elution. The chromatographic analysis revealed a various polyphenols belonging to hydroxycinnamic acid, flavone, flavonol and anthocyanin classes: caffeoylquinic acid, cichoric acid, cyanidin malonylglucoside, kaempferol malonylglucoside, luteolin glucuronide and quercetin malonylglucoside were detected in most samples. The method validation was performed according to ICH guidelines4. The developed analytical method was applied to the Foto di Delunas phytochemical analysis of eight autochthonous C. Cichorium intybus intybus varieties, namely ‘Castelfranco’, ‘Chioggia’,

giugno 2017 natural 1 t 77 ABSTRACT

‘Lusia’, ‘Rosa di Gorizia’, ‘Treviso precoce’, ‘Treviso Ageing represents an important health issue for tardivo’, ‘Verdon’ and ‘Verona’, demonstrating to be modern society. Many studies suggest that ageing a reliable tool for the antioxidant fingerprinting of enhances the vulnerability of several tissue, such as this peculiar endemic salad. brain, liver and heart, against ischemia/reperfusion (I/R) injury. 1.Guarrera, P.M.; Savo, V. Perceived health properties of wild and In this context, sirtuin 1 enzyme (SIRT1) seems to cultivated food plants in local and popular traditions of Italy: A review. J. Ethnopharmacol. 2013, 146, 659–680. play a pivotal role, because of it is involved in the 2.Carazzone, C.; Mascherpa, D.; Gazzani, G.; Papetti, A. Identifi- regulation of many physiological functions and its cation of phenolic constituents in red chicory salads (Cichori- expression decreases with age [1]. In particular, a um intybus) by high-performance liquid chromatography with critical role of SIRT1 in cardioprotection has been diode array detection and electrospray ionisation tandem mass spectrometry. Food Chem. 2013, 138, 1062–1071. reported using SIRT1-deficient and SIRT1-overex- 3.Rossetto, M.; Lante, A.; Vanzani, P.; Spettoli, P.; Scarpa, M.; Rigo, pressing mice [2]; therefore, an effective strategy A. Red chicories as potent scavengers of highly reactive radi- to extend the life-span and to improve tolerance to cals: a study on their phenolic composition and peroxyl radical I/R damage is focused on improving the expression/ trapping capacity and efficiency. J. Agric. Food Chem. 2005, 53, 8169–8175. activity of SIRT1 with exogenous agents, in order 4.International Conference on Harmonization of Technical Re- to modulate different intracellular downstream quirements for the Registration of Pharmaceuticals for Human pathways strictly related with ageing. The poly- Use (ICH). Guidelines Q2 (R1) -Validation of Analytical Proce- dures: Text and Methodology, 2005. ICH Secretariat, c/o IFP- phenol resveratrol represents the lead compound MA, Geneva, 2005, 1–13. of SIRT1-activators but other natural compounds have been reported in the literature, including naringenin (NAR), a flavonoid typical ofCitrus genus [1,3]. Interestingly, NAR conferred cardioprotection in rat myocardium submitted to I/R injury, through the activation of mitochondrial big conductance calcium-activated potassium (mitoBK) channels [4]. Unfortunately, besides SIRT1, an ageing-dependent decline of the mitoBK expression has been observed by us and others [5] and it has been spec- ulated to be a major cause of lesser tolerance of myocardium to I/R injury. This work aims to evaluate the possible anti-ageing effects of NAR, chronically administered to 6 months old male mice. In particular, a first group of animals was treated for 3 months while the second one received NAR for 6 months. At the end of treat- ments, in mice of both the first group (9 months old) and of the second one (12 months old), an im- provement of the mitoBK channel expression was observed. Moreover, as concern SIRT1 enzyme and its down- Fiore di Citrus limon stream pathways, an enhanced expression of SIRT1 accompanied by a marked reduction of the reac- The Citrus flavonoid, tive oxygen species (ROS) production have been naringenin, protects the shown. The effects of NAR-treatment on other age- myocardium against ageing. ing markers correlated with SIRT1, have been also evaluated in order to define the machinery regulat- ed by the Citrus flavonoid. Testai L.1,2, Piragine E.1, Piano I.1, Martelli A.1,2, Citi These preliminary results suggest that a nutraceuti- V. 1, Gargini M.C.1,2, Da Pozzo E. 1,2, Martini C. 1,2, cal approach with NAR may have a positive impact Breschi M.C. 1, Calderone V. 1,2 on critical markers of ageing, and that the use of Citrus flavanone may be suitable for improving the 1 Department of Pharmacy, University of Pisa, Pisa, Italy tolerance of myocardium at I/R damage. 2 Interdepartmental Research Center “Nutraceuticals and Food for Health”, University of Pisa, Pisa, Italy [email protected] Funding: Progetti di Ricerca di Ateneo 2017 (PRA2017).

78 t natural 1 giugno 2017 [1] Meng et al., Cell Mol Neurobiol. 2017 Jan;37(1):17-28. g) were macerated with MeOH-H2O (8:2) at room [2] Nadtochiy et al., Am J Physiol Heart Circ Physiol. 2011 temperature. The residue was suspended in H O Oct;301(4):H1506-12. 2 [3] Howitz et al., Nature. 2003 Sep 11;425(6954):191-6. and successively partitioned with petroleum ether, [4] Testai et al., Biochem Pharmacol. 2013 Jun 1;85(11):1634-43. CHCl3, EtOAc, and n-BuOH affording a CHCl3 solu- [5] Testai et al., Front Pharmacol. 2017 Feb 27;8:71. ble fraction (2 g), an EtOAc-soluble fraction (5.5 g),

and a n-BuOH soluble fraction (19 g). The CHCl3 and n-BuOH extracts were tested for the antioxidant

activity showing radical scavenging ability (CHCl3Ext

DPPH IC50 0.041±0.001 mg/mL, TPC 116.5±0.7

μgGAE/mg; n-BuOHExt DPPH IC50 0.052±0.001 mg/ mL, TPC 94.4±0.9 μgGAE/mg).

Chromatography separation of CHCl3 and n-BuOH extracts led to isolation of 18 pure compounds whose Foto di Alvesgaspar structures were elucidated by 1D-and 2D-NMR spectroscopy and confirmed by mass spectrometry analysis. Sinapic aldehyde, abietin, chlorogenic acid, 5-caffeoyl quinic acid, 8α-hydroxypinoresinol, 9α-hydroxypinoresinol, pinoresinol, 4-ketopinoresinol, nor-trachelogenin, erythro-guaiacylglyc- erol-β-O-4’-dihydroconiferyl alcohol were isolated

from CHCl3 extract; while luteolin 4’-O-glucuronide, naringenin-7-O-neohesperidoside, kaempferol-3-O-a-L-rhamnopyranosyl-(1→6)-b-D-glucopyranoside, kaempferol-7-O-a-L-rhamnopyranosyl-(1→6)-b-D-glucopyranoside, quercitrin, quercetin-3-O-a-l-rhamnopyranosyl-(1→6)-b-D-glucopyra- Galactites noside, ciwujiatone, nor-trachelogenin-4,4’-di-O-β-D- glucopyranoside were obtained from n-BuOH extract. The ability of compounds to reduce the level of per- in-cells antioxidant potential oxides in control and BHP-treated Jurkat cells was of herbal preparations and studied according to Rothe et al.4 components The majority of isolates displayed a significant antiox- from Galactites elegans idant potential in vitro text (DPPH, FRAP, TPC). The lignan derivatives were also able to reduce at 50 μM Roberta Cotugnoa, Omar Tebboubb, Feyza Oke-Al- the basal level of peroxides in Jurkat cells as well as to tuntasc, Mohamed Bouheroumb, Nicola Malafrontea, counteract peroxide increase induced by BHP treat- Fabrizio Dal Piaza, Antonio Vassallod, ment. Particularly 4-ketopinoresinol was the most a active showing 65% of peroxide level inhibition. The Dipartimento di Farmacia, Università di Salerno, Fisciano, slight lower antioxidant potential of nortrachelogenin (SA), Italy. bUnité de Recherche Valorisation des Ressources Naturelles, Fac- 4,4’-di-O-β-D-glucopyranoside (45% of peroxide level ulty of Exact Sciences, Universitè de Constantine1,Algeria inhibition) could be ascribed to the presence of a car- cDepartment of Biology, Faculty of Science, Gazi University, An- bohydrate mojety. kara 06500, Turkey dDipartimeno di Scienze, Università degli Studi della Basilicata, Potenza, (PZ), Italy. References [email protected] 1. Conrad, M. In Methodus Plantas Horti Botanici et Agri Mar- burgensis: a staminum situ describendi. Ed. In officina nova libraria academiae; Marburgi Cattorum, Germany, 1794, Vol. Galactites is a genus of flowering plants belonging 1, p 558. to Asteraceae family. This genus Galactites is main- 2. S. Pignatti. In Flora d’Italia. Ed. Edagricole, Bologna, Italy, ly represented by the Galactites elegans, the milky 1982; Vol. 3, p. 164. 3. Jerković, I.; Roje, M.; Tuberoso, C.I.; Marijanović, Z.; Ka- thistle, a plant of Mediterranean origin (synonym: sum, A.; Obradović, M.; Bioorganic Research of Galactites Galactites tomentosa). tomentosa Moench. Honey Extracts: Enantiomeric Purity of Galactites elegans is consumed as a monofloral boar Chiral Marker 3-Phenyllactic Acid. Chirality. 2014; 26(8), 1-3 405-410. thistle honey. The aerial parts of G. elegans were 4. Rothe, G.; Valet, G. Flow cytometric analysis of respiratory collected in Hamma Bouziane, Constantine, Algeria. burst activity in phagocytes with hydroethidine and 2’, 7’-di- Dried and powdered aerial parts of G. elegans (966 chlorofluorescin. J. leukocyte biol. 1990; 47(5), 440-448.

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