Synergistic antileishmanial activity of erythrodiol, uvaol, and oleanolic acid isolated from olive leaves of cv. Chemlali.

This study aimed to assess the antileishmanial activity of biomolecules obtained from Olea europaea L. leaves and twigs recovered from eight Tunisian cultivars. The extraction was first carried out with 80% methanol, and then the obtained extract was fractionated using three solvents of increasing polarity: cyclohexane (CHX), dichloromethane (DCM) and ethyl acetate (EtOAc). The antileishmanial activity was determined against leishmanial strains responsible for cutaneous, visceral, and mucocutaneous leishmaniasis. The cyclohexane fraction of the leaves of cv. Chemlali from the region of Sidi-Bouzid exhibited the strongest leishmanicidal activity against all the tested leishmanial strains. The inhibition concentrations (IC50) were 16.5, 14.5, and 7.4 µg mL-1 for Leishmania mexicana (cutaneous), Leishmania braziliensis (mucocutaneous), and Leishmania donovani (visceral), respectively. Interestingly, low cytotoxicity was observed on THP-1 cells with selective indexes (SI) ranging from 22.8 to 50.5. HPLC-HRMS and full-house NMR analysis allowed the identification of three triterpenic compounds, oleanolic acid (IC50 = 64.1 µg mL-1), erythrodiol (IC50 = 52.0 µg mL-1), and uvaol (IC50 = 53.8 µg mL-1). Antileishmanial activity of uvaol and oleanolic acid has been previously reported. However, this work constitutes the first report of the antileishmanial activity of erythrodiol which showed combinatorial interaction with uvaol (IC50 = 26.1 µg mL-1) against Leishmania tropica. The mixture of the three compounds, as major ones, exhibited an enhanced activity against Leishmania tropica (IC50 = 16.3 µg mL-1) compared to erythrodiol alone or the combination of uvaol and erythrodiol. This finding is of great importance and needs further investigation. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03825-3.

Bioavailability of Phenolic Compounds in Californian-Style Table Olives with Tunisian Aqueous Olive Leaf Extracts.

Recent advances in biotechnology have ensured that one of the main olive tree by-products is olive leaf extract (OLE), a rich source in bioactive compounds. The aim of this work was to study the phenolic composition in different OLEs of three Tunisian varieties, namely, 'Sayali', 'Tkobri', and 'Neb Jmel'. The in vitro biodigestibility effect after 'Sayali' OLE addition to Californian-style 'Hojiblanca' table olives was also studied. This OLE contained bioactive molecules such as hydroxytyrosol, tyrosol, oleropeine, Procianidine B1 (PB1), and p-cumaric acid. These compounds were also found in fresh olives after OLE was added. Furthermore, from fresh extract to oral digestion, the detected amount of bioavailable phenol was higher; however, its content decreased according to each phase of gastric and intestinal digestion. In the final digestion phase, the number of phenols found was lower than that of fresh olives. In addition, the phenolic content of Californian-style 'Hojiblanca' table olives decreased during the in vitro digestion process. The antioxidant activity of this variety decreased by 64% and 88% after gastrointestinal digestion, being the highest antioxidant capacity found in both simulated gastric and intestinal fluid, respectively. The results show us that the 'Sayali' variety is rich in phenolic compounds that are bioavailable after digestion, which could be used at an industrial level due to the related health benefits.

Evaluation of Tunisian Olive Leaf Extracts to Reduce the Bioavailability of Acrylamide in Californian-Style Black Olives.

The aim of this work was analyzing the use of olive leaf extracts (OLE) obtained from two local Tunisian olive tree cultivars 'Chemlali' and 'Sayali' to reduce the acrylamide in Californian-style black olives. The phenol profile, antioxidant, and antibacterial activity of the two OLE extracts were evaluated. The principal phenols found were hydroxytyrosol (1809.6 ± 25.3 mg 100 g-1), oleuropein (2662.2 ± 38 mg 100 g-1) and luteolin-7-O-glucoside (438.4 ± 38 mg 100 g-1) presented higher levels in 'Sayali' variety. Small differences were observed between the two kinds of extracts used; the greatest activity of OLE was observed against S. choleraesuis, with values up to 50% inhibition. The extract of 'Chemlali' cultivar was added to the Californian-style table olive, improving its phenol content and its antioxidant characteristics without negatively affecting its sensorial characteristics; these olives showed the highest firmness and proper quality characteristics. The gastrointestinal activity on the acrylamide concentration showed a partial degradation of this compound through the digestion, although the addition of the extract does not seem influence in its gastrointestinal digestion. These findings prove the usefulness of by-products to generate a high-quality added-value product, and this would also be relevant as a step towards a more sustainable, circular economy model.

Exogenous nitric oxide alleviates manganese toxicity in bean plants by modulating photosynthesis in relation to leaf lipid composition.

Nitric oxide (NO) is a signaling molecule controlling several steps of plant development and defense process under stress conditions. NO-induced alleviation of manganese (Mn) toxicity was investigated on bean plants submitted for 28 days to 500 µM MnCl2. Manganese excess decreased plant dry weight and elongation and increased levels of reactive oxygen species and lipid peroxidation leading to up-regulation of superoxide dismutase, catalase, and ascorbate peroxidase activities. The inhibitory effects of Mn on plant growth were associated to reduction of light-saturated carbon assimilation (Amax), stomatal conductance (gs), and transpiration (E). By contrast, Mn induced significant increase in the apparent quantum yield (ɸ) and light compensation point (LCP). Interestingly, intracellular CO2 (Ci) remains stable under Mn stress. Concomitantly, leaf membrane lipids have drastically reduced under high Mn concentration. After Mn exposition, leaf fatty acids exhibited a significant loss of linolenic acid, accompanied by an accumulation of palmitoleic, stearic, and linoleic acids leading to alteration of lipid desaturation. NO supply reversed Mn toxicity as evidenced by enhancement of growth biomass and recovery of Amax, E, ɸ, and LCP. Similarly, NO addition has positive effects on leaf lipid content and composition leading to restoration of lipid unsaturation. The modulation of fatty acid composition can be a way to reduce leaf membrane damages and maintain optimal photosynthesis and plant growth. Despite the absence of enough evidences in how NO is involved in lipid and photosynthesis recovery under Mn stress conditions, it is assumed that NO beneficial effects are attributable to NO/Mn cross-talk.

Effectiveness of five flavored Tunisian olive oils on Anisakis larvae type 1: application of cinnamon and rosemary oil in industrial anchovy marinating process.

BACKGROUND: Anisakidosis is caused by the ingestion of raw or undercooked fish or cephalopods containing viable Anisakis larvae. Several natural extracts, oils, essential oils, and their compounds have been tested against Anisakis. In this study the effectiveness of Tunisian olive oil with different spices or plants (cardamom, cinnamon, ginger, laurel, and rosemary) was tested against Anisakis larvae type 1. RESULTS: For the in vitro test, larvae were submerged separately in the oils mentioned above and observed to check viability. Cinnamon oil was the most effective against parasites with lethal time (LT) scores being LT50 = 1.5 days and LT100 = 3 days, followed by rosemary. Laurel, cardamom, and ginger oils were less effective. For the ex vivo experiment, cinnamon, and rosemary oils were tested in anchovy fillets, previously artificially parasitized. Cinnamon was the most effective against parasites (dead after 4 days) as compared to rosemary (7 days). CONCLUSION: The use of cinnamon and rosemary-flavored olive oil in the industrial marinating process can be considered as an efficient alternative to the freezing process required by European Regulation EC No 853/2004 to devitalize Anisakis. © 2019 Society of Chemical Industry.

Changes in phenolic composition in olive tree parts according to development stage.

The aim of this study was to investigate the changes in the phenolic profile in different organs (buds, flowers, fruits and leaves) from Chemlali olive variety cultivated in the Center of Tunisia according to development stage. Phenolics recovery was carried out using aqueous methanol from freeze-dried powdered tissues. The extracts were then analyzed by using high performance liquid chromatography coupled to time of flight and ion trap mass spectrometry detectors. Qualitatively, secoiridoids, flavonoids, simple phenols, cinnamic acid derivatives and lignans were identified in the analyzed extracts. Quantitatively, floral buds showed the highest phenolic contents compared to the other tissues under study. The highest content of secoiridoids was observed for unripe fruit extract, whereas the highest content of flavonoids was registered for floral buds.

LC-MS-based metabolite profiling of methanolic extracts from the medicinal and aromatic species Mentha pulegium and Origanum majorana.

INTRODUCTION: There has been increasing interest dedicated to the phenolic compounds with a view to their antioxidant and healthy properties. Recent studies have focused on plants from the Lamiaceae family with special interest in phenolic compounds antioxidant potential. OBJECTIVE: The metabolite profile of methanolic extracts from two Lamiacea medicinal plants was investigated. MATERIALS AND METHODS: Mentha pulegium and Origanum majorana methanolic extracts were analysed using reversed-phase ultra-high-performance liquid chromatography (RP-UHPLC) coupled to electrospray ionisation quadrupole time-of-flight mass spectrometry (ESI-QTOF-MS) detection in the negative ion mode. RESULTS: A total of 85 metabolites were characterised from different families, such as organic acids and derivatives, amino acids and derivatives, nucleosides, phenolic compounds as well as other polar metabolites, by using the MS and MS/MS information provided by the QTOF-MS. However, the total phenols and flavonoids were also quantified spectrophotometrically and they registered higher amounts in Mentha pulegium than in Origanum majorana extract. Gallocatechin was the major compound in M. pulegium extract whereas quercetin dimethyl ether, jaceidin and dihydrokaempferide were the major ones in O. majorana extract. CONCLUSION: The distribution of phenolic compounds in the methanolic extract showed a variation among studied plants. Mentha pulegium can be considered as a source of gallocatechin.

Olive Tree (Olea europeae L.) Leaves: Importance and Advances in the Analysis of Phenolic Compounds.

Phenolic compounds are becoming increasingly popular because of their potential role in contributing to human health. Experimental evidence obtained from human and animal studies demonstrate that phenolic compounds from Olea europaea leaves have biological activities which may be important in the reduction in risk and severity of certain chronic diseases. Therefore, an accurate profiling of phenolics is a crucial issue. In this article, we present a review work on current treatment and analytical methods used to extract, identify, and/or quantify phenolic compounds in olive leaves.

Characterisation of phenolic compounds by HPLC-TOF/IT/MS in buds and open flowers of 'Chemlali' olive cultivar.

INTRODUCTION: Plant phenolics are secondary metabolites that constitute one of the most widely occurring groups of phytochemicals that play several important functions in plants. In olive (Olea europaea L), there is not enough information about the occurrence of these compounds in buds and flowers. OBJECTIVE: To conduct a comprehensive characterisation of buds and open flowers from the olive cultivar 'Chemlali'. METHODS: The polar fraction of buds and open flowers was extracted using solid-liquid extraction with hydro-alcoholic solvent. Then extracts were analysed using high performance liquid chromatography (HPLC) coupled to electrospray ionisation time-of-flight mass spectrometry (ESI/TOF/MS) and electrospray ionisation ion-trap tandem mass spectrometry (ESI/IT/MS²) operating in negative ion mode. RESULTS: Phenolic compounds from different classes including secoiridoids, flavonoids, simple phenols, cinnamic acid derivatives and lignans were tentatively identified in both extracts. Qualitatively, no significant difference was observed between flower buds and open flowers extracts. However, quantitatively the secoiridoids presented higher percentage of total phenols in open flowers (41.7%) than in flower buds (30.5%) in contrast to flavonoids, which decreased slightly from 38.1 to 26.7%. Cinnamic acid derivatives and simple phenols did not show any change. Lignans presented the lowest percentage in both extracts with an increase during the development of the flower bud to open flower. CONCLUSION: The HPLC-TOF/IT/MS allowed the characterisation, for the first time, of the phenolic profile of extracts of 'Chemlali' olive buds and open flowers, proving to be a very useful technique for the characterisation and structure elucidation of phenolic compounds.

Sterolic composition of Chétoui virgin olive oil: Influence of geographical origin.

The sterol profile of Tunisian virgin olive oils produced from Chétoui cultivar, the second main variety cultivated in the north of the country, grown under different environmental conditions, was established by gas chromatography using a flame ionisation detector. More than ten compounds were identified and characterised. As expected for virgin olive oil, the main sterols found in all Chétoui olive oils were ß-sitosterol, Δ5-avenasterol, campesterol and stigmasterol. Cholesterol, 24-methylenecholesterol, clerosterol, campestanol, sitostanol, Δ7-stigmastenol, Δ5,24-stigmastadienol, and Δ7-avenasterol were also found in all samples, but in lower amounts. Most of these compounds are significantly affected by the geographical origin. The majority of the Chétoui virgin olive oils analysed respected EC Regulation No. 2568, and in all cases total sterol amounts were higher than the minimum limit set by legislation, ranging from 1017 to 1522mg/kg. Two triterpenic dialcohols (erythrodiol and uvaol), were also detected besides the sterolic components. Their content was below the upper legal limit of 4% in all analysed samples, with a range from 1.2% to 3.2%. These results suggest that, besides the genetic factor, environmental conditions influence the sterolic fraction.

Induction of growth inhibition and differentiation of human leukemia HL-60 cells by a Tunisian gerboui olive leaf extract.

Cancer protection associated with the consumption of olive products is well established, but not for leukemia. The protective effects of olive (Olea europaea L.) leaves were investigated by incubating human promyelocytic leukemia HL-60 cells with olive leaf extracts (OLEs) from seven principal Tunisian olive varieties, namely, Chemchali, Chemlali, Chétoui, Gerboui, Sayali, Zalmati and Zarrazi. The results showed significant growth inhibition of HL-60 cells incubated for 48 h with a 100-fold dilution of each OLE which had been obtained by incubating 10 g of dried leaves in 100 ml of 70% ethanol for one week with subsequent ultrafiltration. DNA fragmentation was observed in the cells incubated for 19 h with a 100-fold dilution of the Chemchali, Chemlali and Zalmati extracts. The results of a nitroblue tetrazolium (NBT) assay revealed NBT reduction, a differentiation marker, by the OLE-treated cells after an overnight incubation. The Gerboui extract showed the highest NBT reduction ability at more than 90%. An HPLC analysis revealed the presence of apigenin 7-glucoside in the extract, which was found in subsequent experiments to be responsible for the Gerboui extract-mediated cell differentiation.