Synthesis and Characterization of a Biopolymer Pectin/Ethanolic Extract from Olive Mill Wastewater: In Vitro Safety and Efficacy Tests on Skin Wound Healing.

Wound-healing delay is one of the major problems of type 2 diabetes, representing also a clinical emergency in non-healing chronic wounds. Natural antioxidants show interesting wound-healing properties, including those extracted from waste derived from olive oil production. Olive mill wastewater is one of the main by-products of the olive oil-making process, and it is rich in high-value secondary metabolites, mainly hydroxytyrosol. We proposed an eco-friendly extraction method, employing both ultrasound-assisted and Soxhlet techniques and ethanol as a solvent, to recover valuable molecules from Roggianella cv (Olea europea L.) olive mill wastewater, which was further entrapped in a pectin polymer via an enzymatic reaction using porcine pancreatic lipase. Pectin, in combination with other substances, promoted and accelerated wound healing and demonstrated good potential to produce a biomedical conjugate for wound treatment. The antioxidant activity of the extracts and conjugate were evaluated against lipophilic (IC50 equal to 0.152 mg mL-1) and hydrophilic (IC50 equal to 0.0371 mg mL-1) radical species as well as the in vitro cytotoxicity via NRU, h-CLAT, and a wound-healing scratch assay and assessment. The pectin conjugate did not exert hemolytic effects on the peripheral blood, demonstrating interesting wound-healing properties due to its ability to stimulate cell proliferation in a dose-dependent manner.

Phenolic Extract from Olive Leaves as a Promising Endotherapeutic Treatment against Xylella fastidiosa in Naturally Infected Olea europaea (var. europaea) Trees.

(1) Background: Since 2013, the pathogenic bacterium Xylella fastidiosa has been severely affecting olive production in Apulia, Italy, with consequences for the economy, local culture, landscape and biodiversity. The production of a phenolic extract from fresh olive leaves was employed for endotherapeutic injection into naturally infected olive trees by Xylella fastidiosa in Apulia region, Italy. (2) Methods: The effectiveness of the extract was tested in vitro and in planta in comparison with analogous treatments based on garlic powder and potassium phosphite. (3) Results: The uptake of phenolic compounds from olive leaves through a trunk injection system device resulted in a statistically significant increase in leaf area index and leaf area density, as well as in the growth of newly formed healthy shoots. Plant growth-promoting effects were also observed for potassium phosphite. Moreover, the bacteriostatic activities of the phenolic extract and of the garlic-powder-based solution have been demonstrated in in vitro tests. (4) Conclusions: The results obtained and the contained costs of extraction make the endotherapeutic treatment with phenolic compounds a promising strategy for controlling X fastidiosa to be tested on a larger scale, although the experiments conducted in this study proved not to be suitable for centenary trees.

Olea europaea L. Flowers as a new promising anticancer natural product: phenolic composition, antiproliferative activity and apoptosis induction.

The aim of this study was to characterise phenolic compounds of olive flower obtained from Olive tree cultivar Chemlali and to investigate their anticancer effect on MCF-7 cells. Phenolic characterisation was determined using LC/MS-MS. Cytotoxicity of the extract was determined using MTT. Biochemical markers of apoptosis were evaluated by immunoblotting. Our results showed that olive flower contained significant amounts of phenolic compounds mainly flavonoids, secoiridoids and simple phenols. Furthermore, the phenolic extract exerted a significant reduction in MCF-7 cell viability (EC50 values equal to 220.8 µg/ml). Western blot analysis revealed the presence of the cleaved forms of Parp-1. The DAPI staining analysis demonstrated a significant reduction in the number of cells and a considerable change in the morphology of the treated cells. In conclusion, Olea europaea. L flower contained great amounts of different bio-phenols able to reduce the proliferative activity of breast cancer MCF-7 cells by the induction of apoptosis.

Eco-Friendly Extraction and Characterisation of Nutraceuticals from Olive Leaves.

Olive tree (Olea europaea L.) leaf, a waste by-product of the olive oil industry, is an inexpensive and abundant source of biophenols of great interest for various industrial applications in the food supplement, cosmetic, and pharmaceutical industries. In this work, the aqueous extraction of high-added value compounds from olive leaves by using microfiltered (MF), ultrapure (U), and osmosis-treated (O) water was investigated. The extraction of target compounds, including oleuropein (Olp), hydroxytyrosol (HyTyr), tyrosol (Tyr), verbascoside (Ver), lutein (Lut), and rutin (Rut), was significantly affected by the characteristics of the water used. Indeed, according to the results of liquid chromatography tandem mass spectrometry, the extracting power of microfiltered water towards rutin resulted very poor, while a moderate extraction was observed for oleuropein, verbascoside, and lutein. On the other hand, high concentrations of hydroxytyrosol were detected in the aqueous extracts produced with microfiltered water. The extraction power of ultrapure and osmosis-treated water proved to be very similar for the bio-active compounds oleuropein, verbascoside, lutein, and rutin. The results clearly provide evidence of the possibility of devising new eco-friendly strategies based on the use of green solvents which can be applied to recover bioactive compounds from olive leaves.

Elemental Characterization of Medicinal Plants and Soils from Hazarganji Chiltan National Park and Nearby Unprotected Areas of Balochistan, Pakistan.

The aim of this work was to evaluate the variability in elemental composition of seven medicinal plants and their respective soils belonging to protected and nearby unprotected sites of the Hazarganji Chiltan National Park. The medical plants under study were Achillea wilhelmsii C. Koch, Peganum harmala Linn, Sophora mollis (Royle) Baker, Perovskia atriplicifolia Benth, Seriphidium quettense (Podlech.) Ling, Hertia intermedia (Bioss) O. Ktze, and Nepeta praetervisa Rech. F. Macro (C, H, N, S, K, Ca), micro (Cl, Cu, Fe, Mn, and Zn), beneficial (Al, Co, Na), others (As, Br, Cr, Cs, Hf, Pb, Rb, Sb, Sr, Sn, V and Th) and rare earth elements (Ce, Eu, La, Lu, Nd Sc, Sm, Tb and Yb) were characterized by means of standard organic elemental and instrumental neutron activation methodologies and by flame atomic absorption spectroscopy. Results showed that, among macro nutrients, carbon concentration was the highest element in both plant and soil samples followed by H and K. Elements such as Cl, Na and Fe were detected in considerably good amounts; all the other elements were found in trace quantities. Principal component analysis (PCA) was applied to identify spatial variation in elemental composition of medicinal plants, in which 80-90% of the total variance in whole set of data was found. In particular, the findings highlighted the presence of essential and beneficial elements such as C, H, N, K, Ca, Fe, Mn and Na, in samples from protected sites, while potentially dangerous elements such as Al, As, Br and Cr were detected in samples from unprotected sites. These results emphasized on the need for rational exploitation of valuable medicinal plants and supporting protected areas as an excellent source of biodiversity conservation.

Quality and trace element profile of Tunisian olive oils obtained from plants irrigated with treated wastewater.

In the present work the use of treated wastewater (TWW) to irrigate olive plants was monitored. This type of water is characterized by high salinity and retains a substantial amount of trace elements, organic and metallic compounds that can be transferred into the soil and into the plants and fruits. In order to evaluate the impact of TWW on the overall quality of the oils, the time of contact of the olives with the soil has been taken into account. Multi-element data were obtained using ICP-MS. Nineteen elements (Li, B, Na, Mg, Al, K, Ca, Sc, Cr, Mn, Fe, Co, Ni, Cu, Zn, Sr, Mo, Ba and La) were submitted for statistical analysis. Using analysis of variance, linear discriminant analysis and principal component analysis it was possible to differentiate between oils produced from different batches of olives whose plants received different types of water. Also, the results showed that there was correlation between the elemental and mineral composition of the water used to irrigate the olive plots and the elemental and mineral composition of the oils.

Determination of trace element in Italian virgin olive oils and their characterization according to geographical origin by statistical analysis.

Multi-element analysis of organic virgin olive oils from different Italian regions was carried out by inductively coupled plasma mass spectrometry (ICP-MS) aiming at developing a reliable method in the traceability of the origin of oils. The data were processed by means of the chemiometric approach of linear discriminant analysis (LDA) that allows classifying unknown samples after checking possible differentiation of samples of known origin. An external calibration curve was build for the quantitative analysis. The calibration curves for each element were linear in the range between 0.01 and 100 ng mL(-1) and 0.2 to 2000 ng mL(-1), the correlation coefficients were ranging between 0.996 and 0.999. Results from spike and recovery experiments at levels of 30 and 65 ng mL(-1) were in the range of 91-119%, whereas the quantitation limits, based on 10 times standard deviation of the blank, were also in the range of 0.009-10.2 ng g(-1), for almost all the elements.

Assay of aroma active components of virgin olive oils from southern Italian regions by SPME-GC/ion trap mass spectrometry.

An SPME-GC/ion trap method was exploited to determine the chromatogram of volatile compounds of organic olive oils of southern Italian regions. The method is based on the assay of the terminal species of the "lipoxygenase pathway", which are present in the volatile fraction of the sampled compounds. Ethyl isobutanoate was used as internal standard in either the EI or CI ionization mode. The absolute concentration values of each analyte were evaluated through good-to-excellent calibration curves. Case studies on oils obtained from different cultivars or harvesting times are presented. The quantitative data for each compound were subjected to principal component analysis to characterize the different cultivars of this work.