Cytotoxic Effect Induced by Sicilian Oregano Essential Oil in Human Breast Cancer Cells.

Origanum vulgare L. is an aromatic plant that exerts antibacterial, antioxidant, anti-inflammatory, and antitumor activities, mainly due to its essential oil (EO) content. In this study, we investigated the possible mechanism underlying the in vitro antitumor activity of EO extracted by hydrodistillation of dried flowers and leaves of Origanum vulgare L. grown in Sicily (Italy) in MDA-MB-231 and MCF-7 breast cancer cell lines. Gas chromatography-mass spectrometry analysis of Oregano essential oil (OEO) composition highlighted the presence of twenty-six major phytocompounds, such as p-cymene, γ-terpinene, and thymoquinone p-acetanisole. OEO possesses strong antioxidant capacity, as demonstrated by the DPPH test. Our studies provided evidence that OEO reduces the viability of both MCF-7 and MDA-MB-231 cells. The cytotoxic effect of OEO on breast cancer cells was partially counteracted by the addition of z-VAD-fmk, a general caspase inhibitor. Caspases and mitochondrial dysfunction appeared to be involved in the OEO-induced death mechanism. Western blotting analysis showed that OEO-induced activation of pro-caspases-9 and -3 and fragmentation of PARP decreased the levels of Bcl-2 and Bcl-xL while increasing those of Bax and VDAC. In addition, fluorescence microscopy and cytofluorimetric analysis showed that OEO induces a loss of mitochondrial membrane potential in both cell lines. Furthermore, we tested the effects of p-cymene, γ-terpinene, thymoquinone, and p-acetanisole, which are the main components of OEO. Our findings highlighted that the effect of OEO on MDA-MB-231 and MCF-7 cells appears to be mainly due to the combination of different constituents of OEO, providing evidence of the potential use of OEO for breast cancer treatment.

Impact of Ecklonia maxima Seaweed Extract and Mo Foliar Treatments on Biofortification, Spinach Yield, Quality and NUE.

Seaweed extract (SE) application is a contemporary and sustainable agricultural practice used to improve yield and quality of vegetable crops. Plant biofortification with trace element is recognized as a major tool to prevent mineral malnourishment in humans. Mo deficiency causes numerous dysfunctions, mostly connected to central nervous system and esophageal cancer. The current research was accomplished to appraise the combined effect of Ecklonia maxima brown seaweed extract (SE) and Mo dose (0, 0.5, 2, 4 or 8 µmol L-1) on yield, biometric traits, minerals, nutritional and functional parameters, as well as nitrogen indices of spinach plants grown in a protected environment (tunnel). Head fresh weight (FW), ascorbic acid, polyphenols, N, P, K, Mg and nitrogen use efficiency (NUE) were positively associated with SE treatment. Moreover, head FW, head height (H), stem diameter (SD), ascorbic acid, polyphenols, carotenoids as well as NUE indices were enhanced by Mo-biofortification. A noticeable improvement in number of leaves (N. leaves), head dry matter (DM) and Mo concentration in leaf tissues was observed when SE application was combined with a Mo dosage of 4 or 8 µmol L-1. Overall, our study highlighted that E. maxima SE treatment and Mo supply can improve both spinach production and quality via the key enzyme activity involved in the phytochemical homeostasis of SE and the plant nutritional status modification resulting in an enhanced spinach Mo tolerance.

Effects of Irrigation with Different Sources of Water on Growth, Yield and Essential Oil Compounds in Oregano.

Aromatic plants can benefit from the use of treated wastewater to satisfy their water requirements, but the effects on the essential oil yield and quality need an assessment. The aims of this study were to assess the effects of freshwater and treated wastewater obtained from a Sicilian (Italy) pilot-scale horizontal subsurface flow constructed wetland system on plant growth and yield, essential oil yield and composition of oregano (Origanum vulgare ssp. hirtum (Link) Ietswaart) and soil characteristics. The system had a total surface area of 100 m2 and was planted with giant reed and umbrella sedge. An experimental open field of oregano was set up close to the system. Two years and two different sources of irrigation water were tested in a split-plot design for a two-factor experiment. Treated wastewater was characterized by higher values of mineral and organic constituents than freshwater. The results highlight that short-term irrigation with freshwater and treated wastewater, in both years, led to increased plant growth, dry weight and essential oil yield of oregano plants. However, it did not significantly affect the essential oil content and composition in comparison with the control. Furthermore, the year and source of irrigation water did not significantly vary the chemical composition of the soil. Our results suggest that treated wastewater can be considered an alternative to freshwater for the cultivation of oregano due to the fact that it does not greatly influence the yield quality and quantity of this species in the short-term.

A Two-Year Bio-Agronomic and Chemotaxonomic Evaluation of Wild Sicilian Myrtle (Myrtus communis L.) Berries and Leaves.

A collection of nine Myrtus communis samples from different localities of Sicily was evaluated. Morphological traits and production characteristics have been chosen as parameters to arrange the samples into homogeneous groups and to identify the best biotypes for possible future agro-industrial exploitation. The plant material has been subjected to taxonomic characterization from biometric and phytochemical perspectives. Myrtle berries and leaves have been analyzed for their content in metabolites, applying a cascade extraction protocol for M. communis leaves and a single hydroalcoholic extraction for berries, whereas hydrodistillation procedures have been applied to obtain the essential oils from berries and leaves. The analyses of non-volatile components were carried out by LC-UV-DAD-ESI-MS. All the extracts were characterized by the presence of numerous polyphenols, namely highly hydroxylated flavonols such as quercetin and myricetin; and ellagic acid detected in all samples. In addition, myrtle berries contained nine different anthocyanins, namely delphinidin, petunidin, cyanidin and malvidin derivatives. The essential oils (EOs) were analyzed by a combination of GC-FID and GC/MS. A total of 33 and 34 components were fully characterized with the predominance of α-pinene, myrtenyl acetate, linalool, 1,8-cineole and linalyl acetate. All phytochemical profiles were subjected to cluster analyses, which allowed subdividing the myrtle samples in different chemical groups.

Phytochemical, Ecological and Antioxidant Evaluation of Wild Sicilian Thyme: Thymbra capitata (L.) Cav.

In a broad survey conducted throughout the Sicily region, 45 different sites were identified where thyme grows wild. All the biotypes collected were classified as Thymbra capitata (L.) Cav. (syn. Thymus capitatus (L.) Hoffmanns. & Link). Cluster analysis based on the main morphological characteristics of the plant led to the division of the biotypes into 3 major groups. All samples were analyzed for their secondary phytochemical metabolites identified in the extracts and the essential oils. LC-UV-DAD/ESI-MS and GC-FID/GC-MS have been applied to characterize the extracts and the essential oils, respectively. In the extracts, 15 flavonoid derivatives with taxifolin-di-O-glucoside and thymusin as main components, and 2 organic acids, with a large predominance of rosmarinic acid, were identified. On the whole 37 compounds were fully characterized in the essential oils, carvacrol was identified as the main component with an average value of 73.93%. The total phenol content and the antioxidant activity of all phytochemical complexes were determined with the Folin-Ciocalteu (FC) assay, the UV radiation-induced peroxidation in liposomal membranes (UV-IP test), and the scavenging activity of superoxide radical (O2∙-).

Composition and Variability of the Essential Oil of the Flowers of Lavandula stoechas from Various Geographical Sources.

Samples of flowers of wild Lavandula stoechas L. spp. stoechas populations were collected in three areas of Sicily (Italy) and were characterized in agronomic and chemical terms. Essential oil (EO) was extracted by hydrodistillation and analyzed by GC-FID and GC-MS. GC-FID and GC-MS analyses permitted identification of 89 compounds from the EO. The samples were separated into 3 groups using PCA (Principal Component Analysis) statistical method, with reference to the chemical composition of the EO. All three Sicilian populations of lavender were identified as the fenchone chemotype with percentage content ranged between 52.8-71.1%. The population of Partinico showed the highest dry weight of flowers per plant (221.3 g), but the lowest EO yield (0.37%). The essential oils of the three wild Sicilian populations of L. stoechas L. spp. stoechas showed a greater chemical differentiation than those obtained from other Mediterranean areas.

Biomolecular characterization of wild sicilian oregano: phytochemical screening of essential oils and extracts, and evaluation of their antioxidant activities.

An extensive survey of wild Sicilian oregano was made. A total of 57 samples were collected from various sites, followed by taxonomic characterization from an agronomic perspective. Based on morphological and production characteristics obtained from the 57 samples, cluster analysis was used to divide the samples into homogeneous groups, to identify the best biotypes. All samples were analyzed for their phytochemical content, applying a cascade-extraction protocol and hydrodistillation, to obtain the non volatile components and the essential oils, respectively. The extracts contained thirteen polyphenol derivatives, i.e., four flavanones, seven flavones, and two organic acids. Their qualitative and quantitative characterization was carried out by LC/MS analyses. The essential oils were characterized using a combination of GC-FID and GC/MS analyses; a total of 81 components were identified. The major components of the oils were thymol, p-cymene, and γ-terpinene. Cluster analysis was carried out on both phytochemical profiles and resulted in the division of the oregano samples into different chemical groups. The antioxidant activity of the essential oils and extracts was investigated by the Folin-Ciocalteau (FC) colorimetric assay, by UV radiation-induced peroxidation in liposomal membranes (UV-IP test), and by determining the O(2)(∙-)-scavenging activity.