Daniele Fraternale - Academia.edu (original) (raw)

Papers by Daniele Fraternale

Research paper thumbnail of Cytotoxic activity of essential oils of aerial parts and ripe fruits of Echinophora spinosa (Apiaceae)

Natural Product Communications, 2013

The cytotoxic effects of the essential oils obtained from the flowering aerial parts (APO) and ri... more The cytotoxic effects of the essential oils obtained from the flowering aerial parts (APO) and ripe fruits (RFO) of Echinophora spinosa L. (Apiaceae) from central Italy toward human U937 promonocytoid cells were studied; the contribution of each of the major constituents to the whole cytotoxic activity of either APO or RFO was also characterized. The major components of APO were beta-phellandrene (34.7%), myristicin (16.5%), p-cymene (16.3%), delta3-carene (12.6%), alpha-pinene (6.7%) and alpha-phellandrene (6.2%); those of RFO p-cymene (50.2%), myristicin (15.3%), alpha-pinene (15.1%) and alpha-phellandrene (8.1%). Both oils tested were toxic to U937 cells, but RFO was much more cytotoxic: indeed, the IC50 values calculated from the linear regression curves of RFO and APO were 14.5 +/- 0.85 and 43.4 +/- 2.81 microg/mL, respectively. alpha-Pinene and alpha-phellandrene were identified as the most toxically relevant constituents: however, they did not completely account for the toxic...

Research paper thumbnail of Prunus spinosa Extract Loaded in Biomimetic Nanoparticles Evokes In Vitro Anti-Inflammatory and Wound Healing Activities

Nanomaterials, 2020

Prunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inf... more Prunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inflammatory activities. Innovative drug delivery systems such as biomimetic nanoparticles could improve the activity of PSF extract by promoting (i) the protection of payload into the lipidic bilayer, (ii) increased accumulation to the diseased tissue due to specific targeting properties, (iii) improved biocompatibility, (iv) low toxicity and increased bioavailability. Using membrane proteins extracted from human monocyte cell line THP-1 cells and a mixture of phospholipids, we formulated two types of PSF-extract-loaded biomimetic vesicles differing from each other for the presence of either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DOPG). The biological activity of free extract (PSF), compared to both types of extract-loaded vesicles (PSF-DOPCs and PSF-DOPGs) and empty vesicles (DOPCs and DOPGs), was evaluated in vitro on HUVEC...

Research paper thumbnail of Ferulago campestris Essential Oil as Active Ingredient in Chitosan Seed-Coating: Chemical Analyses, Allelopathic Effects, and Protective Activity against the Common Bean Pest Acanthoscelides obtectus

Ferulago campestris is an aromatic plant, distributed in Mediterranean Europe, and a source of es... more Ferulago campestris is an aromatic plant, distributed in Mediterranean Europe, and a source of essential oils (EOs) regarded as promising alternatives to synthetic herbicides and pesticides. F. campestris EO, hydrodistilled and analyzed, was tested for its antigerminative activity and for its effect on radicle elongation, hydrogen peroxide concentration, and lipid peroxidation on four infesting weed species (Papaver rhoeas, Taraxacum campylodes, Poa annua, and Setaria verticillata) and on Phaseolus vulgaris. Its repellent ability was also tested against Acanthoscelides obtectus, a pest commonly affecting P. vulgaris during storage. Moreover, a chitosan coating layer was developed with the addition of F. campestris EO and tested for its toxic and oviposition-deterring effects against A. obtectus. Myrcene, α-pinene, and γ-terpinene were detected as the main compounds in F. campestris EO. The EO demonstrated a selective in vitro antigerminative activity towards the weed species, withou...

Research paper thumbnail of Yield, Characterization, and Possible Exploitation of Cannabis Sativa L. Roots Grown under Aeroponics Cultivation

Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and ps... more Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C. sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, β-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth—particularly of the roots—and a significant increase (up to 20-fold in the case of β-sitosterol) in the total content of the aforementioned roots’ bioactive molecules we...

Research paper thumbnail of Bioconversion of Callus-Produced Precursors to Silymarin Derivatives in Silybum marianum Leaves for the Production of Bioactive Compounds

The present study aimed to investigate the enzymatic potential of Silybum marianum leaves to bioc... more The present study aimed to investigate the enzymatic potential of Silybum marianum leaves to bioconvert phenolic acids produced in S. marianum callus into silymarin derivatives as chemopreventive agent. Here we demonstrate that despite the fact that leaves of S. marianum did not accumulate silymarin themselves, expanding leaves had the full capacity to convert di-caffeoylquinic acid to silymarin complex. This was proven by HPLC separations coupled with electrospray ionization mass spectrometry (ESI-MS) analysis. Soaking the leaf discs with S. marianum callus extract for different times revealed that silymarin derivatives had been formed at high yield after 16 h. Bioconverted products displayed the same retention time and the same mass spectra (MS or MS/MS) as standard silymarin. Bioconversion was achieved only when using leaves of a specific age, as both very young and old leaves failed to produce silymarin from callus extract. Only medium leaves had the metabolic capacity to conver...

Research paper thumbnail of Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production

International Journal of Molecular Sciences, 2021

Genetic engineering of plants has turned out to be an attractive approach to produce various seco... more Genetic engineering of plants has turned out to be an attractive approach to produce various secondary metabolites. Here, we attempted to produce kynurenine, a health-promoting metabolite, in plants of Nicotiana tabacum (tobacco) transformed by Agrobacterium tumefaciens with the gene, coding for human indoleamine 2,3-dioxygenase 1 (IDO1), an enzyme responsible for the kynurenine production because of tryptophan degradation. The presence of IDO1 gene in transgenic plants was confirmed by PCR, but the protein failed to be detected. To confer higher stability to the heterologous human IDO1 protein and to provide a more sensitive method to detect the protein of interest, we cloned a gene construct coding for IDO1-GFP. Analysis of transiently transfected tobacco protoplasts demonstrated that the IDO1-GFP gene led to the expression of a detectable protein and to the production of kynurenine in the protoplast medium. Interestingly, the intracellular localisation of human IDO1 in plant cell...

Research paper thumbnail of Antioxidant and Anti-Inflammaging Ability of Prune (Prunus Spinosa L.) Extract Result in Improved Wound Healing Efficacy

Antioxidants, 2021

Prunus spinosa L. fruit (PSF) ethanol extract, showing a peculiar content of biologically active ... more Prunus spinosa L. fruit (PSF) ethanol extract, showing a peculiar content of biologically active molecules (polyphenols), was investigated for its wound healing capacity, a typical feature that declines during aging and is negatively affected by the persistence of inflammation and oxidative stress. To this aim, first, PSF anti-inflammatory properties were tested on young and senescent LPS-treated human umbilical vein endothelial cells (HUVECs). As a result, PSF treatment increased miR-146a and decreased IRAK-1 and IL-6 expression levels. In addition, the PSF antioxidant effect was validated in vitro with DPPH assay and confirmed by in vivo treatments in C. elegans. Our findings showed beneficial effects on worms’ lifespan and healthspan with positive outcomes on longevity markers (i.e., miR-124 upregulation and miR-39 downregulation) as well. The PSF effect on wound healing was tested using the same cells and experimental conditions employed to investigate PSF antioxidant and anti-i...

Research paper thumbnail of A New Strategy to Increase Production of Genoprotective Bioactive Molecules from Cotyledon-Derived Silybum marianum L. Callus

Genes, 2020

There is a need to enhance the production of bioactive secondary metabolites and to establish new... more There is a need to enhance the production of bioactive secondary metabolites and to establish new production systems, e.g., for liver-protective compounds of Silybum marianum seeds. Quantifying and identifying the produced phytochemicals, and examining their protective effects against genotoxic agents, is of great interest. This study established a protocol for the qualitative and quantitative production of hepatoprotective compounds in cotyledon-derived Silybum marianum callus through optimized supplementation of the MS medium with the growth regulators 2,4-D, benzylaminopurine, myoinositol, and asparagine. High-performance liquid chromatography (HPLC) coupled with electrospray ionisation mass spectrometry (ESI-MS) allowed for identification and quantification of the produced compounds. None of the growth medium combinations supported a detectable production of silymarin. Instead, the generated calli accumulated phenolic acids, in particular chlorogenic acid and dicaffeoylquinic ac...

Research paper thumbnail of Bioeffects of Prunus spinosa L. fruit ethanol extract on reproduction and phenotypic plasticity of Trichoplax adhaerens Schulze, 1883 (Placozoa)

PeerJ, 2019

The aim of this work was to test and analyse the bioeffects of Prunus spinosa L. (Rosacaee) fruit... more The aim of this work was to test and analyse the bioeffects of Prunus spinosa L. (Rosacaee) fruit ethanol extract on Trichoplax adhaerens Schulze, 1883 (Placozoa) laboratory cultures which—for the first time—were employed as in vivo biological model to assess the bioactivity of a natural extract. The ethanol extract of P. spinosa was administrated during a 46 day experimental period; ultrastructural (by optical, confocal, TEM and SEM microscopy) and morphometric analyses indicated that treated Trichoplax adhaerens showed significant differences in viability, reproductive modalities, body shape and colour with respect to the control group. Finally, P. spinosa bioactive compounds seem to exert profound protective effects on T. adhaerens reproduction and phenotype. Our results may support additional investigations related to other bioactive compounds properties useful for nutraceutical preparations to be used as food supplements.

Research paper thumbnail of Essential Oil Composition and Antigermination Activity of Artemisia Dracunculus (Tarragon)

Natural Product Communications, 2015

The chemical composition of an Italian oil of tarragon ( Artemisia dracunculus L.) was analyzed b... more The chemical composition of an Italian oil of tarragon ( Artemisia dracunculus L.) was analyzed by GC/EIMS. The major compound of the oil was estragole (73.3%), followed by limonene (5.4%), ( E)-β-ocimene (5.3%), β-pinene (3.4%) and ( Z)-β-ocimene (3.0%). The essential oil was tested “ in vitro” for its antigermination activity against Raphanus sativus L., Lepidium sativum L, Papaver rhoeas L. and Avena fatua L. seeds and demonstrating a good inhibitory activity in a dose-dependent way.

Research paper thumbnail of In Vitro Culture for Ex Situ Conservation of “Roveja” and Nutritional Considerations on this Italian Rare Legume

Natural Product Communications, 2018

Roveja is a pea cultivar from the Sibillini Mountains (Umbria-Marche Apennine mountain range, Ita... more Roveja is a pea cultivar from the Sibillini Mountains (Umbria-Marche Apennine mountain range, Italy). Abandoned for long time, Roveja is nowadays cultivated only in few farms. The present work deals with the in vitro plant growth of this pea by tissue culture for shoots regeneration and ex situ conservation. Total polyphenols content evaluation, DPPH (2,2-diphenyl-1-picrylhydrazyl) antioxidant activity test and ORAC (Oxygen Radical Absorbance Capacity) assay were performed on the flour obtained from Roveja seeds and the results were compared to those collected by performing the same tests on the flour from two common green peas. The best shoots regeneration from embryonic axes was obtained in Murashige & Skoog (MS) media with combinations of 4.0 mg/L BA (6-Benzyladenine) plus 0.4 mg/L NAA (1-Naphthaleneacetic acid), and the best roots induction from the regenerated shoots was obtained in half strength MS media containing 1.0 mg/L IBA (3-Indolebutyric acid). The embryonic axes of Rov...

Research paper thumbnail of Activity of Vitis vinifera Tendrils Extract Against Phytopathogenic Fungi

Natural Product Communications, 2015

The in vitro antifungal activity was determined of an ethanolic extract of Vitis vinifera L. tend... more The in vitro antifungal activity was determined of an ethanolic extract of Vitis vinifera L. tendrils (TVV) against ten plant pathogenic fungi, using the agar dilution method; activity was shown against all tested fungi. Fusarium species were the most sensitive with MIC values ranging from 250 to 300 ppm, while the basidiomycete fungus Rhizoctonia solani was the most resistant, with a MIC value of 500 ppm. Electrospray ionization tandem mass spectrometry (ESI-MSn) was used to obtain qualitative information on the main components of TVV. The high amount of polyphenolic compounds contained in TVV is likely to contribute significantly to its antifungal activity.

Research paper thumbnail of Composition and Antioxidant Activity of Inula crithmoides Essential Oil Grown in Central Italy (Marche Region)

Natural Product Communications, 2010

The chemical composition of the essential oil obtained from the aerial parts of Inula crithmoides... more The chemical composition of the essential oil obtained from the aerial parts of Inula crithmoides L. was analyzed by GC and GC/MS and 22 components were identified, the major ones being p-cymene (30.1%), 1-methylethyl-trimethylbenzene (18.7%), scopoletin (15.3%) and α-pinene (13.1%). The antioxidant activity of the oil was evaluated by the DPPH test and 5-lipoxygenase assay. The essential oil exerted a good antioxidant activity in the protection of lipid peroxidation when compared with known antioxidants.

Research paper thumbnail of Lipoxygenase Inhibitory Activity of Boropinic Acid, Active Principle of Boronia Pinnata

Natural Product Communications, 2006

Boropinic acid and other natural prenyloxycinnamic and benzoic acids were easily synthesized in h... more Boropinic acid and other natural prenyloxycinnamic and benzoic acids were easily synthesized in high yield by a two-step sequence from the corresponding p-hydroxy aromatic acids and were assayed for radical scavenging activity using the DPPH test and for inhibition of enzymatic lipid peroxidation mediated by soybean 5-lipoxygenase. Compared to other acids and to known antioxidant compounds like BHT, Trolox and ascorbic acid, boropinic acid was far more active in the lipoxygenase test (IC50 = 7.6 ng/mL, p < 0.05). The recorded inhibition value suggested that boropinic acid acted as an enzyme inhibitor rather than a mere radical or peroxide scavenger. This hypothesis was confirmed by studying the interaction between boropinic acid and soybean 5-lipoxygenase by molecular modelling techniques.

Research paper thumbnail of Antifungal Evaluation of Hypericum Triquetrifolium Polar Extracts against Fusarium spp

Natural Product Communications, 2006

The chemical composition of different kinds of Hypericum triquetrifolium extracts was analyzed by... more The chemical composition of different kinds of Hypericum triquetrifolium extracts was analyzed by LC-DAD-ESI-MS. Hyperoside, rutin, isoquercitrin and biapigenin were the main constituents. As these natural compounds were reported in the literature for their antifungal activity, the total extracts were tested for their antifungal activity against eight phytopathogenic strains of Fusarium species.

Research paper thumbnail of Aqueous Extract from Vitis vinifera Tendrils is Able to Enrich Keratinocyte Antioxidant Defences

Natural Product Communications, 2011

An aqueous extract of V. vinifera L. tendrils was evaluated for its ability to enrich the antioxi... more An aqueous extract of V. vinifera L. tendrils was evaluated for its ability to enrich the antioxidant capacity of cultured cells. The long-time antioxidant capability of the extract was measured by in vitro chemical methods, and its influence on reduced glutathione levels and plasma membrane oxido reductase activity was determined in cultured human keratinocytes (NCTC 2544). Keratinocytes are cells normally exposed to oxidative stress, and for this reason adequately equipped with antioxidant defences. However, it has long been suggested that exogenous antioxidants may play an important role in minimizing the adverse effects of oxidative stress on skin. We demonstrated that V. vinifera tendril aqueous extract was able to increase, in a time- and dose-dependent manner, the reduced glutathione concentration and activity of trans plasma membrane oxido reductase as an indirect evaluation of the intracellular redox status of the cells demonstrating a relevant antioxidant activity of this ...

Research paper thumbnail of Phytotoxic Activity of Salvia x jamensis

Natural Product Communications, 2009

A study has been carried out on the surface exudate of Salvia x jamensis, which showed a signific... more A study has been carried out on the surface exudate of Salvia x jamensis, which showed a significant phytotoxic activity against Papaver rhoeas L. and Avena sativa L.. Bioguided separation of the exudate yielded active fractions from which 3β-hydroxy-isopimaric acid (1), hautriwaic acid (2), betulinic acid (3), 7,8β-dihydrosalviacoccin (4), isopimaric acid (5), 14α-hydroxy-isopimaric acid (7), 15,16-epoxy-7α,10β-dihydroxy-clerod-3,13(16),14-trien-17,12;18,19-diolide (8), cirsiliol (5,3′,4′-trihydroxy-6,7-dimethoxyflavone, 9) and two new neoclerodane diterpenes (6 and 10) were isolated. The structures of 6 and 10 were identified as 15,16-epoxy-10β-hydroxy-clerod-3,13(16),14-trien-17,12;18,19-diolide and 15,16-epoxy-7α,10-dihydroxy-clerod-2,13(16),14-trien-17,12;18,19-diolide respectively on the basis of spectroscopic data analysis. All compounds, but 7, 8 and 10, were active in inhibiting the germination of the tested species.

Research paper thumbnail of The Essential Oil of Monarda didyma L. (Lamiaceae) Exerts Phytotoxic Activity in Vitro against Various Weed Seed

Molecules (Basel, Switzerland), Jan 2, 2017

The chemical composition of the essential oil of the flowering aerial parts of Monarda didyma L. ... more The chemical composition of the essential oil of the flowering aerial parts of Monarda didyma L. cultivated in central Italy was analyzed by Gas Chromatography/Mass Spectrometry (GC/MS). The major compounds of the oil were thymol (59.3%), p-cymene (10.3%), terpinolene (9.2%), δ-3-carene (4.4%), myrcene (3.7%), and camphene (3.4%). The essential oil was tested in vitro for its anti-germination activity against Papaver rhoeas L., Taraxacum officinale F. H. Wigg., Avena fatua L., Raphanus sativus L. and Lepidium sativum L. seeds, demonstrating good inhibitory activity in a dose-dependent way. The exposure of the employed weed seeds to M. didyma essential oil and thymol solution (59.3%) increased the level of hydrogen peroxide (H2O2) and malondialdehyde (MDA), markers of oxidative stress, in emerging 5-day-old rootlets.

Research paper thumbnail of Essential oil composition of Angelica archangelica L. (Apiaceae) roots and its antifungal activity against plant pathogenic fungi

Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology, 2014

Research paper thumbnail of Control of isolates of Fusarium species by using extracts of Rubus ulmifolius micropropagated plantlets

We developed a micropropagation protocol to obtain in vitro plantlets from nodal explants of Rubu... more We developed a micropropagation protocol to obtain in vitro plantlets from nodal explants of Rubus ulmifolius. The methanolic extract of dried plantlets was assayed for antifungal activity on the growth of eight isolates of Fusarium species. The molecular components of the extract, separated by column chromatography, were analysed by HPLC. Rubus ulmifolius micropropagated plantlets provide uniformly conditioned material available all year around. Their phytochemicals are characterized by a high antioxidant capacity, water-solubility and ability to inhibit the growth of the eight isolates of Fusarium species.

Research paper thumbnail of Cytotoxic activity of essential oils of aerial parts and ripe fruits of Echinophora spinosa (Apiaceae)

Natural Product Communications, 2013

The cytotoxic effects of the essential oils obtained from the flowering aerial parts (APO) and ri... more The cytotoxic effects of the essential oils obtained from the flowering aerial parts (APO) and ripe fruits (RFO) of Echinophora spinosa L. (Apiaceae) from central Italy toward human U937 promonocytoid cells were studied; the contribution of each of the major constituents to the whole cytotoxic activity of either APO or RFO was also characterized. The major components of APO were beta-phellandrene (34.7%), myristicin (16.5%), p-cymene (16.3%), delta3-carene (12.6%), alpha-pinene (6.7%) and alpha-phellandrene (6.2%); those of RFO p-cymene (50.2%), myristicin (15.3%), alpha-pinene (15.1%) and alpha-phellandrene (8.1%). Both oils tested were toxic to U937 cells, but RFO was much more cytotoxic: indeed, the IC50 values calculated from the linear regression curves of RFO and APO were 14.5 +/- 0.85 and 43.4 +/- 2.81 microg/mL, respectively. alpha-Pinene and alpha-phellandrene were identified as the most toxically relevant constituents: however, they did not completely account for the toxic...

Research paper thumbnail of Prunus spinosa Extract Loaded in Biomimetic Nanoparticles Evokes In Vitro Anti-Inflammatory and Wound Healing Activities

Nanomaterials, 2020

Prunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inf... more Prunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inflammatory activities. Innovative drug delivery systems such as biomimetic nanoparticles could improve the activity of PSF extract by promoting (i) the protection of payload into the lipidic bilayer, (ii) increased accumulation to the diseased tissue due to specific targeting properties, (iii) improved biocompatibility, (iv) low toxicity and increased bioavailability. Using membrane proteins extracted from human monocyte cell line THP-1 cells and a mixture of phospholipids, we formulated two types of PSF-extract-loaded biomimetic vesicles differing from each other for the presence of either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DOPG). The biological activity of free extract (PSF), compared to both types of extract-loaded vesicles (PSF-DOPCs and PSF-DOPGs) and empty vesicles (DOPCs and DOPGs), was evaluated in vitro on HUVEC...

Research paper thumbnail of Ferulago campestris Essential Oil as Active Ingredient in Chitosan Seed-Coating: Chemical Analyses, Allelopathic Effects, and Protective Activity against the Common Bean Pest Acanthoscelides obtectus

Ferulago campestris is an aromatic plant, distributed in Mediterranean Europe, and a source of es... more Ferulago campestris is an aromatic plant, distributed in Mediterranean Europe, and a source of essential oils (EOs) regarded as promising alternatives to synthetic herbicides and pesticides. F. campestris EO, hydrodistilled and analyzed, was tested for its antigerminative activity and for its effect on radicle elongation, hydrogen peroxide concentration, and lipid peroxidation on four infesting weed species (Papaver rhoeas, Taraxacum campylodes, Poa annua, and Setaria verticillata) and on Phaseolus vulgaris. Its repellent ability was also tested against Acanthoscelides obtectus, a pest commonly affecting P. vulgaris during storage. Moreover, a chitosan coating layer was developed with the addition of F. campestris EO and tested for its toxic and oviposition-deterring effects against A. obtectus. Myrcene, α-pinene, and γ-terpinene were detected as the main compounds in F. campestris EO. The EO demonstrated a selective in vitro antigerminative activity towards the weed species, withou...

Research paper thumbnail of Yield, Characterization, and Possible Exploitation of Cannabis Sativa L. Roots Grown under Aeroponics Cultivation

Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and ps... more Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C. sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, β-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth—particularly of the roots—and a significant increase (up to 20-fold in the case of β-sitosterol) in the total content of the aforementioned roots’ bioactive molecules we...

Research paper thumbnail of Bioconversion of Callus-Produced Precursors to Silymarin Derivatives in Silybum marianum Leaves for the Production of Bioactive Compounds

The present study aimed to investigate the enzymatic potential of Silybum marianum leaves to bioc... more The present study aimed to investigate the enzymatic potential of Silybum marianum leaves to bioconvert phenolic acids produced in S. marianum callus into silymarin derivatives as chemopreventive agent. Here we demonstrate that despite the fact that leaves of S. marianum did not accumulate silymarin themselves, expanding leaves had the full capacity to convert di-caffeoylquinic acid to silymarin complex. This was proven by HPLC separations coupled with electrospray ionization mass spectrometry (ESI-MS) analysis. Soaking the leaf discs with S. marianum callus extract for different times revealed that silymarin derivatives had been formed at high yield after 16 h. Bioconverted products displayed the same retention time and the same mass spectra (MS or MS/MS) as standard silymarin. Bioconversion was achieved only when using leaves of a specific age, as both very young and old leaves failed to produce silymarin from callus extract. Only medium leaves had the metabolic capacity to conver...

Research paper thumbnail of Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production

International Journal of Molecular Sciences, 2021

Genetic engineering of plants has turned out to be an attractive approach to produce various seco... more Genetic engineering of plants has turned out to be an attractive approach to produce various secondary metabolites. Here, we attempted to produce kynurenine, a health-promoting metabolite, in plants of Nicotiana tabacum (tobacco) transformed by Agrobacterium tumefaciens with the gene, coding for human indoleamine 2,3-dioxygenase 1 (IDO1), an enzyme responsible for the kynurenine production because of tryptophan degradation. The presence of IDO1 gene in transgenic plants was confirmed by PCR, but the protein failed to be detected. To confer higher stability to the heterologous human IDO1 protein and to provide a more sensitive method to detect the protein of interest, we cloned a gene construct coding for IDO1-GFP. Analysis of transiently transfected tobacco protoplasts demonstrated that the IDO1-GFP gene led to the expression of a detectable protein and to the production of kynurenine in the protoplast medium. Interestingly, the intracellular localisation of human IDO1 in plant cell...

Research paper thumbnail of Antioxidant and Anti-Inflammaging Ability of Prune (Prunus Spinosa L.) Extract Result in Improved Wound Healing Efficacy

Antioxidants, 2021

Prunus spinosa L. fruit (PSF) ethanol extract, showing a peculiar content of biologically active ... more Prunus spinosa L. fruit (PSF) ethanol extract, showing a peculiar content of biologically active molecules (polyphenols), was investigated for its wound healing capacity, a typical feature that declines during aging and is negatively affected by the persistence of inflammation and oxidative stress. To this aim, first, PSF anti-inflammatory properties were tested on young and senescent LPS-treated human umbilical vein endothelial cells (HUVECs). As a result, PSF treatment increased miR-146a and decreased IRAK-1 and IL-6 expression levels. In addition, the PSF antioxidant effect was validated in vitro with DPPH assay and confirmed by in vivo treatments in C. elegans. Our findings showed beneficial effects on worms’ lifespan and healthspan with positive outcomes on longevity markers (i.e., miR-124 upregulation and miR-39 downregulation) as well. The PSF effect on wound healing was tested using the same cells and experimental conditions employed to investigate PSF antioxidant and anti-i...

Research paper thumbnail of A New Strategy to Increase Production of Genoprotective Bioactive Molecules from Cotyledon-Derived Silybum marianum L. Callus

Genes, 2020

There is a need to enhance the production of bioactive secondary metabolites and to establish new... more There is a need to enhance the production of bioactive secondary metabolites and to establish new production systems, e.g., for liver-protective compounds of Silybum marianum seeds. Quantifying and identifying the produced phytochemicals, and examining their protective effects against genotoxic agents, is of great interest. This study established a protocol for the qualitative and quantitative production of hepatoprotective compounds in cotyledon-derived Silybum marianum callus through optimized supplementation of the MS medium with the growth regulators 2,4-D, benzylaminopurine, myoinositol, and asparagine. High-performance liquid chromatography (HPLC) coupled with electrospray ionisation mass spectrometry (ESI-MS) allowed for identification and quantification of the produced compounds. None of the growth medium combinations supported a detectable production of silymarin. Instead, the generated calli accumulated phenolic acids, in particular chlorogenic acid and dicaffeoylquinic ac...

Research paper thumbnail of Bioeffects of Prunus spinosa L. fruit ethanol extract on reproduction and phenotypic plasticity of Trichoplax adhaerens Schulze, 1883 (Placozoa)

PeerJ, 2019

The aim of this work was to test and analyse the bioeffects of Prunus spinosa L. (Rosacaee) fruit... more The aim of this work was to test and analyse the bioeffects of Prunus spinosa L. (Rosacaee) fruit ethanol extract on Trichoplax adhaerens Schulze, 1883 (Placozoa) laboratory cultures which—for the first time—were employed as in vivo biological model to assess the bioactivity of a natural extract. The ethanol extract of P. spinosa was administrated during a 46 day experimental period; ultrastructural (by optical, confocal, TEM and SEM microscopy) and morphometric analyses indicated that treated Trichoplax adhaerens showed significant differences in viability, reproductive modalities, body shape and colour with respect to the control group. Finally, P. spinosa bioactive compounds seem to exert profound protective effects on T. adhaerens reproduction and phenotype. Our results may support additional investigations related to other bioactive compounds properties useful for nutraceutical preparations to be used as food supplements.

Research paper thumbnail of Essential Oil Composition and Antigermination Activity of Artemisia Dracunculus (Tarragon)

Natural Product Communications, 2015

The chemical composition of an Italian oil of tarragon ( Artemisia dracunculus L.) was analyzed b... more The chemical composition of an Italian oil of tarragon ( Artemisia dracunculus L.) was analyzed by GC/EIMS. The major compound of the oil was estragole (73.3%), followed by limonene (5.4%), ( E)-β-ocimene (5.3%), β-pinene (3.4%) and ( Z)-β-ocimene (3.0%). The essential oil was tested “ in vitro” for its antigermination activity against Raphanus sativus L., Lepidium sativum L, Papaver rhoeas L. and Avena fatua L. seeds and demonstrating a good inhibitory activity in a dose-dependent way.

Research paper thumbnail of In Vitro Culture for Ex Situ Conservation of “Roveja” and Nutritional Considerations on this Italian Rare Legume

Natural Product Communications, 2018

Roveja is a pea cultivar from the Sibillini Mountains (Umbria-Marche Apennine mountain range, Ita... more Roveja is a pea cultivar from the Sibillini Mountains (Umbria-Marche Apennine mountain range, Italy). Abandoned for long time, Roveja is nowadays cultivated only in few farms. The present work deals with the in vitro plant growth of this pea by tissue culture for shoots regeneration and ex situ conservation. Total polyphenols content evaluation, DPPH (2,2-diphenyl-1-picrylhydrazyl) antioxidant activity test and ORAC (Oxygen Radical Absorbance Capacity) assay were performed on the flour obtained from Roveja seeds and the results were compared to those collected by performing the same tests on the flour from two common green peas. The best shoots regeneration from embryonic axes was obtained in Murashige & Skoog (MS) media with combinations of 4.0 mg/L BA (6-Benzyladenine) plus 0.4 mg/L NAA (1-Naphthaleneacetic acid), and the best roots induction from the regenerated shoots was obtained in half strength MS media containing 1.0 mg/L IBA (3-Indolebutyric acid). The embryonic axes of Rov...

Research paper thumbnail of Activity of Vitis vinifera Tendrils Extract Against Phytopathogenic Fungi

Natural Product Communications, 2015

The in vitro antifungal activity was determined of an ethanolic extract of Vitis vinifera L. tend... more The in vitro antifungal activity was determined of an ethanolic extract of Vitis vinifera L. tendrils (TVV) against ten plant pathogenic fungi, using the agar dilution method; activity was shown against all tested fungi. Fusarium species were the most sensitive with MIC values ranging from 250 to 300 ppm, while the basidiomycete fungus Rhizoctonia solani was the most resistant, with a MIC value of 500 ppm. Electrospray ionization tandem mass spectrometry (ESI-MSn) was used to obtain qualitative information on the main components of TVV. The high amount of polyphenolic compounds contained in TVV is likely to contribute significantly to its antifungal activity.

Research paper thumbnail of Composition and Antioxidant Activity of Inula crithmoides Essential Oil Grown in Central Italy (Marche Region)

Natural Product Communications, 2010

The chemical composition of the essential oil obtained from the aerial parts of Inula crithmoides... more The chemical composition of the essential oil obtained from the aerial parts of Inula crithmoides L. was analyzed by GC and GC/MS and 22 components were identified, the major ones being p-cymene (30.1%), 1-methylethyl-trimethylbenzene (18.7%), scopoletin (15.3%) and α-pinene (13.1%). The antioxidant activity of the oil was evaluated by the DPPH test and 5-lipoxygenase assay. The essential oil exerted a good antioxidant activity in the protection of lipid peroxidation when compared with known antioxidants.

Research paper thumbnail of Lipoxygenase Inhibitory Activity of Boropinic Acid, Active Principle of Boronia Pinnata

Natural Product Communications, 2006

Boropinic acid and other natural prenyloxycinnamic and benzoic acids were easily synthesized in h... more Boropinic acid and other natural prenyloxycinnamic and benzoic acids were easily synthesized in high yield by a two-step sequence from the corresponding p-hydroxy aromatic acids and were assayed for radical scavenging activity using the DPPH test and for inhibition of enzymatic lipid peroxidation mediated by soybean 5-lipoxygenase. Compared to other acids and to known antioxidant compounds like BHT, Trolox and ascorbic acid, boropinic acid was far more active in the lipoxygenase test (IC50 = 7.6 ng/mL, p < 0.05). The recorded inhibition value suggested that boropinic acid acted as an enzyme inhibitor rather than a mere radical or peroxide scavenger. This hypothesis was confirmed by studying the interaction between boropinic acid and soybean 5-lipoxygenase by molecular modelling techniques.

Research paper thumbnail of Antifungal Evaluation of Hypericum Triquetrifolium Polar Extracts against Fusarium spp

Natural Product Communications, 2006

The chemical composition of different kinds of Hypericum triquetrifolium extracts was analyzed by... more The chemical composition of different kinds of Hypericum triquetrifolium extracts was analyzed by LC-DAD-ESI-MS. Hyperoside, rutin, isoquercitrin and biapigenin were the main constituents. As these natural compounds were reported in the literature for their antifungal activity, the total extracts were tested for their antifungal activity against eight phytopathogenic strains of Fusarium species.

Research paper thumbnail of Aqueous Extract from Vitis vinifera Tendrils is Able to Enrich Keratinocyte Antioxidant Defences

Natural Product Communications, 2011

An aqueous extract of V. vinifera L. tendrils was evaluated for its ability to enrich the antioxi... more An aqueous extract of V. vinifera L. tendrils was evaluated for its ability to enrich the antioxidant capacity of cultured cells. The long-time antioxidant capability of the extract was measured by in vitro chemical methods, and its influence on reduced glutathione levels and plasma membrane oxido reductase activity was determined in cultured human keratinocytes (NCTC 2544). Keratinocytes are cells normally exposed to oxidative stress, and for this reason adequately equipped with antioxidant defences. However, it has long been suggested that exogenous antioxidants may play an important role in minimizing the adverse effects of oxidative stress on skin. We demonstrated that V. vinifera tendril aqueous extract was able to increase, in a time- and dose-dependent manner, the reduced glutathione concentration and activity of trans plasma membrane oxido reductase as an indirect evaluation of the intracellular redox status of the cells demonstrating a relevant antioxidant activity of this ...

Research paper thumbnail of Phytotoxic Activity of Salvia x jamensis

Natural Product Communications, 2009

A study has been carried out on the surface exudate of Salvia x jamensis, which showed a signific... more A study has been carried out on the surface exudate of Salvia x jamensis, which showed a significant phytotoxic activity against Papaver rhoeas L. and Avena sativa L.. Bioguided separation of the exudate yielded active fractions from which 3β-hydroxy-isopimaric acid (1), hautriwaic acid (2), betulinic acid (3), 7,8β-dihydrosalviacoccin (4), isopimaric acid (5), 14α-hydroxy-isopimaric acid (7), 15,16-epoxy-7α,10β-dihydroxy-clerod-3,13(16),14-trien-17,12;18,19-diolide (8), cirsiliol (5,3′,4′-trihydroxy-6,7-dimethoxyflavone, 9) and two new neoclerodane diterpenes (6 and 10) were isolated. The structures of 6 and 10 were identified as 15,16-epoxy-10β-hydroxy-clerod-3,13(16),14-trien-17,12;18,19-diolide and 15,16-epoxy-7α,10-dihydroxy-clerod-2,13(16),14-trien-17,12;18,19-diolide respectively on the basis of spectroscopic data analysis. All compounds, but 7, 8 and 10, were active in inhibiting the germination of the tested species.

Research paper thumbnail of The Essential Oil of Monarda didyma L. (Lamiaceae) Exerts Phytotoxic Activity in Vitro against Various Weed Seed

Molecules (Basel, Switzerland), Jan 2, 2017

The chemical composition of the essential oil of the flowering aerial parts of Monarda didyma L. ... more The chemical composition of the essential oil of the flowering aerial parts of Monarda didyma L. cultivated in central Italy was analyzed by Gas Chromatography/Mass Spectrometry (GC/MS). The major compounds of the oil were thymol (59.3%), p-cymene (10.3%), terpinolene (9.2%), δ-3-carene (4.4%), myrcene (3.7%), and camphene (3.4%). The essential oil was tested in vitro for its anti-germination activity against Papaver rhoeas L., Taraxacum officinale F. H. Wigg., Avena fatua L., Raphanus sativus L. and Lepidium sativum L. seeds, demonstrating good inhibitory activity in a dose-dependent way. The exposure of the employed weed seeds to M. didyma essential oil and thymol solution (59.3%) increased the level of hydrogen peroxide (H2O2) and malondialdehyde (MDA), markers of oxidative stress, in emerging 5-day-old rootlets.

Research paper thumbnail of Essential oil composition of Angelica archangelica L. (Apiaceae) roots and its antifungal activity against plant pathogenic fungi

Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology, 2014

Research paper thumbnail of Control of isolates of Fusarium species by using extracts of Rubus ulmifolius micropropagated plantlets

We developed a micropropagation protocol to obtain in vitro plantlets from nodal explants of Rubu... more We developed a micropropagation protocol to obtain in vitro plantlets from nodal explants of Rubus ulmifolius. The methanolic extract of dried plantlets was assayed for antifungal activity on the growth of eight isolates of Fusarium species. The molecular components of the extract, separated by column chromatography, were analysed by HPLC. Rubus ulmifolius micropropagated plantlets provide uniformly conditioned material available all year around. Their phytochemicals are characterized by a high antioxidant capacity, water-solubility and ability to inhibit the growth of the eight isolates of Fusarium species.