Olive leaf was first used medicinally in Ancient Egypt.

Contents

• Botanical classification of olive leaf.
• Phytochemicals and Constituents:Mechanisms.
• Brief Benefits of Olive Leaf Extract.
• Olive Leaf Extract:the Microbe Fighter:Olive Leaf Extract Natural Defense against Biowarfare Antibacterial Agent and Inactivating Viral Invaders.
• Research Progress and identified Applicaitons Olive Leaves.
• Therapeutic Applications and Administration.
• Olive Leaf Extract as a new formulation choice.
• History of The Olive Tree.
• A Simple Combination:Echinacea-Goldenseal-Olive leaf Formula.
• Olive Exorcism.
• Olive Leaf:Research Update.

Olive Leaf:Research Update.:

  Antioxidant capacity of extracts from wild and crop plants of the Mediterranean region.:J Food Sci. 2007 Jan;72(1):S059-63.Mohamed R, Pineda M, Aguilar M.Dept. de Bioquímica y Biología Molecular, Univ. de Córdoba. Campus de Rabanales, Spain.

 There is an increasing demand for natural antioxidants to replace synthetic additives in the food industry. The present work examines the potential of some wild and cultivated plants from the Mediterranean region as sources of natural antioxidants. Samples of different organs and tissues from each of these species were extracted with aqueous and organic solvents and analyzed for their total hydrosoluble and lipid-soluble antioxidant capacity, measured by the phosphomolybdenum method, and for their content in tocopherols. Our results demonstrate that there is more than a 1000-fold difference among total antioxidants in various plant sources. The highest level of water-soluble antioxidant capacity was found in fruit peel and leaf samples, while seeds presented the highest levels of lipid-soluble antioxidant capacity. Tocopherols were more abundant in leaf samples. Holm oak and olive tree leaves showed the highest yields of alpha-tocopherol. These Mediterranean plant species could be used as a very good source of both water-soluble and lipid-soluble antioxidants, particularly alpha-tocopherol.

  Retention and distribution of polyphenols after pan-frying of French fries in oils enriched with olive leaf extract.:J Food Sci. 2007 Oct;72(8):S574-84.Chiou A, Salta FN, Kalogeropoulos N, Mylona A, Ntalla I, Andrikopoulos NK.Laboratory of Chemistry, Biochemistry, Physical Chemistry of Foods, Dept. of Science of Dietetics-Nutrition, Harokopio Univ., 70 El. Venizelou Ave., 176 71 Kallithea, Athens, Greece

 Palm oil, olive oil, and sunflower oil were supplemented with an extract rich in polyphenols obtained from olive tree (Olea europaea) leaves at levels of 120 and 240 mg total polyphenols per kilogram of oil. Pan-frying of potatoes was performed in both the enriched and the nonsupplemented oils under domestic frying conditions. Total polyphenol content was estimated by the Folin-Ciocalteau assay, oleuropein was determined by HPLC analysis, while other individual polyphenols by GC/MS analysis. Fourteen polyphenol species were identified in the olive leaf extract, among which oleuropein predominated (1.25 g/kg olive leaves). All the enriched oils contained oleuropein before and after frying. Oleuropein as well as other polyphenol species were detected in all French fries cooked in enriched oils. Polyphenol intake by consuming French fries pan-fried in the enriched oils was calculated to be 6 to 31 times higher than that in the case of French fries fried in commercial oils, being dependent on the frying oil type.

  Phenolic compounds and antimicrobial activity of olive (Olea europaea L. Cv. Cobran?osa) leaves.:Molecules. 2007 May 26;12(5):1153-62.Pereira AP, Ferreira IC, Marcelino F, Valent?o P, Andrade PB, Seabra R, Estevinho L, Bento A, Pereira JA.CIMO/Escola Superior Agrária, Instituto Politécnico de Bragan?a, Campus Sta Apolónia, Apt. 1172, 5301-855 Bragan?a, Portugal.

 We report the determination of phenolic compounds in olive leaves by reversed-phase HPLC/DAD, and the evaluation of their in vitro activity against several microorganisms that may be causal agents of human intestinal and respiratory tract infections, namely gram positive (Bacillus cereus, B. subtilis and Staphylococcus aureus), gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae) and fungi (Candida albicans and Cryptococcus neoformans). Seven phenolic compounds were identified and quantified: caffeic acid, verbascoside, oleuropein, luteolin 7-O-glucoside, rutin, apigenin 7-O-glucoside and luteolin 4'-O-glucoside. At low concentrations olive leaves extracts showed an unusual combined antibacterial and antifungal action, which suggest their great potential as nutraceuticals, particularly as a source of phenolic compounds.

  Encapsulation of olive leaf extract in beta-cyclodextrin:J Agric Food Chem. 2007 Oct 3;55(20):8088-94. Epub 2007 Sep 1.Mourtzinos I, Salta F, Yannakopoulou K, Chiou A, Karathanos VT.Laboratory of Chemistry-Biochemistry-Physical Chemistry of Foods, Department of Nutrition & Dietetics, Harokopio University, El. Venizelou 70., Kallithea, 176 71 Athens, Greece.

 Olive leaf extract, rich in oleuropein, formed an inclusion complex with beta-cyclodextrin (beta-CD) upon mixing of the components in aqueous media and subsequent freeze-drying. Inclusion complex formation was confirmed by differential scanning calorimetry (DSC). DSC thermograms indicated that the endothermic peaks of both the olive leaf extract and the physical mixture of olive leaf extract with beta-CD, attributed to the melting of crystals of the extract, were absent in DSC thermogram of inclusion complex. Moreover, DSC studies under oxidative conditions indicated that the complex of olive leaf extract with beta-CD was protected against oxidation, since it remained intact at temperatures where the free olive leaf extract was oxidized. Phase solubility studies afforded A L type diagrams, 1:1 complex stoichiometry, a moderate binding constant ( approximately 300 M (-1)), and an increase of the aqueous solubility by approximately 50%. The formation of the inclusion complex was also confirmed by nuclear magnetic resonance (NMR) studies of beta-CD solutions in the presence of both pure oleuropein and olive leaf extract. The NMR data have established the formation of a 1:1 complex with beta-CD that involves deep insertion of the dihydroxyphenethyl moiety inside the cavity from its secondary side.

  Ultrasound-assisted extraction and silylation prior to gas chromatography-mass spectrometry for the characterization of the triterpenic fraction in olive leaves.:J Chromatogr A. 2007 Sep 21;1165(1-2):158-65. Epub 2007 Jul 24.Sánchez Avila N, Priego Capote F, Luque de Castro MD.Department of Analytical Chemistry, Annex C-3 Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain.

 One of the most important fractions of bioactive compounds isolated from plants is that formed by triterpenic compounds, which have proved to be anti-bacterial, antifungal, anti-inflammatory, cytotoxic and anti-tumour. A method for leaching and determination of the main triterpenic compounds (oleanolic acid, ursolic acid, uvaol, erythrodiol) in olive leaves is here presented. Quantitative leaching was obtained with ethanol as leachant and ultrasonic assistance for 20 min, a very short time as compared to conventional procedures by maceration, which usually requires at least 5 h. After isolation, an aliquot of the ethanolic leachate was silylated to derivatize the analytes prior to gas chromatography-mass spectrometry analysis. Silylation reaction was also assisted with ultrasound in order to accelerate the derivatization step, which only required 5 min--a dramatic shortening in comparison to conventional silylation of terpenic compounds with derivatization times ranging from 30 min to 3 h. The proposed method has demonstrated to be useful for isolation and characterization of the triterpenic fraction in plants; the capability of ultrasound to assist sample preparation (acceleration of leaching and derivatization) has also been proved.


  Irrigation effects on quality, phenolic composition, and selected volatiles of virgin olive oils cv. Leccino.:J Agric Food Chem. 2007 Aug 8;55(16):6609-18. Epub 2007 Jul 18.Servili M, Esposto S, Lodolini E, Selvaggini R, Taticchi A, Urbani S, Montedoro G, Serravalle M, Gucci R.Dipartimento di Scienze Economico-Estimative e degli Alimenti, Sezione di Tecnologie e Biotecnologie degli Alimenti, Università degli Studi di Perugia, Via S. Costanzo, 06126 Perugia, Italy. servimau@unipg.it

 Field-grown olive trees (Olea europaea L. cv. Leccino) were used over two growing seasons to determine the effect of deficit irrigation regimes on virgin olive oil (VOO) quality. Drip irrigation was managed to maintain a predawn leaf water potential (PLWP): (a) higher than -1.1 MPa (full irrigation: FI); (b) between -1.0 and -3.3 MPa (deficit irrigation: DI); (c) higher than -4.2 MPa (severe deficit irrigation: SI). The fruit yield and oil yield of DI trees were over 90% of those of FI treatments in both years, respectively, whereas yields of SI trees ranged from 61 to 76%. The irrigation regime had minor effects on the free acidity, peroxide value, and fatty acid composition of VOO. The concentrations of phenols and o-diphenols in VOO were negatively correlated with PLWP. The concentrations of the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol (3,4-DHPEA-EDA), the isomer of the oleuropein aglycon (3,4-DHPEA-EA), and the dialdehydic form of decarboxymethyl elenolic acid linked to (p-hydroxyphenyl)ethanol (p-HPEA-EDA) were lower in FI than in SI treatments. The concentrations of lignans (+)-1-acetoxipinoresinol and (+)-1-pinoresinol were unaffected by the irrigation regime. The tree water status had a marked effect on the concentration of volatile compounds, such as the C(6)-saturated and unsaturated aldehydes, alcohols, and esters.

  Triterpenes in the hexane extract of leaves of Olea europaea L.: analysis using 13C-NMR spectroscopy.:Phytochem Anal. 2007 Jul;18(4):347-53.Duquesnoy E, Castola V, Casanova J.Université de Corse, Equipe Chimie et Biomasse, UMR CNRS 6134, Route des Iles Sanguinaire, 20000 Ajaccio, France.

 Two neutral triterpenes and a triterpene acid were identified and quantified directly, in the absence of any purification steps, in a precipitate obtained during the industrial extraction of the leaves of Olea europaea L. using 13C-NMR spectroscopy (spectrometer operating at 4.7 T equipped with a 10 mm probe). The method was optimised in order to reduce the duration of analysis with a routine NMR spectrometer. Together with long-chain linear compounds, erythrodiol, uvaol and oleanolic acid accounted for 27.3, 18.3 and 12.5% of the precipitate, respectively.

  Effects of inert dust on olive (Olea europaea L.) leaf physiological para.:Environ Sci Pollut Res Int. 2007 May;14(3):212-4.Nanos GD, Ilias IF.School of Agricultural Science, University of Thessaly, Fitoko Str, 38446 Volos, Greece. gnanos@agr.uth.gr

 BACKGROUND: Cement factories are major pollutants for the surrounding areas. Inert dust deposition has been found to affect photosynthesis, stomatal functioning and productivity. Very few studies have been conducted on the effects of cement kiln dust on the physiology of perennial fruit crops. Our goal was to study some cement dust effects on olive leaf physiology.effects on olive leaf physiology. On METHODS: Cement kiln dust has been applied periodically since April 2003 onto olive leaves. Cement dust accumulation and various leaf physiological parameters were evaluated early in July 2003. Measurements were also taken on olive trees close to the cement factory. RESULTS: Leaf dry matter content and specific leaf weight increased with leaf age and dust content. Cement dust decreased leaf total chlorophyll content and chlorophyll a/chlorophyll b ratio. As a result, photosynthetic rate and quantum yield decreased. In addition, transpiration rate slightly decreased, stomatal conductance to H2O and CO2 movement decreased, internal CO2 concentration remained constant and leaf temperature increased. DISCUSSION: The changes in chlorophyll are possibly due to shading and/or photosystem damage. The changes in stomatal functioning were possibly due to dust accumulation between the peltates or othe effects on stomata. CONCLUSIONS: Dust (in this case from a cement kiln) seems to cause substantial changes to leaf physiology, possibly leading to reduced olive productivity. RECOMMENDATIONS: Avoidance of air contamination from cement factories by using available technology should be examined together with any possible methodologies to reduce plant tissue contamination from cement dust. PERSPECTIVES: Longterm effects of dust (from cement kiln or other sources) on olive leaf, plant productivity and nutritional quality of edible parts could be studied for conclusive results on dust contamination effects to perennial crops.

  Studies on the lipophilicity of vehicles (or co-vehicles) and botanical oils used in cosmetic products.:Pharmazie. 2007 May;62(5):351-3.Mbah CJ.DMI Inc., Wharton, NJ, USA.

 The lipophilic character of five vehicles (or co-vehicles): diethylhexylmaleate, dimethicone, light mineral oil, octyldodecanol and oleyl alcohol and eight botanical oils: Aloe vera oil, coconut oil, extra virgin olive oil, grape leaf oil, grape seed oil, hazelnut oil, jojoba oil and safflower oil was determined by partitioning esters of p-hydroxybenzoic acid (parabens) between them and phosphate buffer (pH 7.4). The results were compared to those obtained with 1-octanol. The most lipophilic effects were observed with octyldodecanol and oleyl alcohol for the vehicles (or co-vehicles), coconut oil, jojoba oil and safflower oil for botanical oils. Light mineral oil showed the least lipophilic effect. With butylparaben, it was observed that oleyl alcohol, octyldodecanol, coconut oil and jojoba oil were 0.94, 0.91, 0.74 and 0.68 times as lipophilic as 1-octanol respectively. The study indicates that octyldodecanol and oleyl alcohol could be good substitutes for 1-octanol in partition coefficient determination. The estimated permeability coefficients of the parabens suggest that octyldodecanol, oleyl alcohol, coconut oil and jojoba oil could be potential dermal permeation enhancers.

  Computational study of bindings of olive leaf extract (OLE) to HIV-1 fusion protein gp41.:FEBS Lett. 2007 Jun 12;581(14):2737-42. Epub 2007 May 21. Bao J, Zhang DW, Zhang JZ, Huang PL, Huang PL, Lee-Huang S.Department of Chemistry, New York University, New York, NY 10003, USA.

 Recent experimental study found that OLE (olive leaf extract) has anti-HIV activity by blocking the HIV virus entry to host cells [Lee-Huang, S., Zhang, L., Huang, P.L., Chang, Y. and Huang, P.L. (2003) Anti-HIV activity of olive leaf extract (OLE) and modulation of host cell gene expression by HIV-1 infection and OLE treatment. Biochem. Biophys. Res. Commun. 307, 1029; Lee-Huang, S., Huang, P.L., Zhang, D., Lee, J.W., Bao, J., Sun, Y., Chang, Y.-Tae, Zhang, J.Z.H. and Huang, P.L. (2007) Discovery of small-molecule HIV-1 fusion and integrase inhibitors oleuropein and hydroxytyrosol. Biochem. Biophys. Res. Commun. 354, 872-878, 879-884]. As part of a joint experimental and theoretical effort, we report here computational study to help identify and characterize the binding complexes of several main compounds of OLE (olive leaf extract) to HIV-1 envelop protein gp41. A number of possible binding modes are found by docking oleuropein and its metabolites, aglycone, elenolic acid and hydroxytyrosol, onto the hydrophobic pocket on gp41. Detailed OLE-gp41 binding interactions and free energies of binding are obtained through molecular dynamics simulation and MM-PBSA calculation. Specific molecular interactions in our predicted OLE/gp41 complexes are identified and hydroxytyrosol is identified to be the main moiety for binding to gp41. This computational study complements the corresponding experimental investigation and helps establish a good starting point for further refinement of OLE-based gp41 inhibitors.


  The olive constituent oleuropein exhibits proteasome stimulatory properties in vitro and confers life span extension of human embryonic fibroblasts.:Rejuvenation Res. 2007 Jun;10(2):157-72. Katsiki M, Chondrogianni N, Chinou I, Rivett AJ, Gonos ES.Institute of Biological Research and Biotechnology, Laboratory of Molecular and Cellular Aging, National Hellenic Research Foundation, Athens, Greece.

 Normal human fibroblasts undergo replicative senescence due to both genetic and environmental factors. Senescence and aging can be further accelerated by exposure of cells to a variety of oxidative agents that contribute among other effects to the accumulation of damaged proteins. The proteasome, a multicatalytic nonlysosomal protease, has impaired function during aging, while its increased expression delays senescence in human fibroblasts. The aim of this study was to identify natural compounds that enhance proteasome activity and exhibit antiaging properties. We demonstrate that oleuropein, the major constituent of Olea europea leaf extract, olive oil and olives, enhances the proteasome activities in vitro stronger than other known chemical activators, possibly through conformational changes of the proteasome. Moreover, continuous treatment of early passage human embryonic fibroblasts with oleuropein decreases the intracellular levels of reactive oxygen species (ROS), reduces the amount of oxidized proteins through increased proteasome-mediated degradation rates and retains proteasome function during replicative senescence. Importantly, oleuropein-treated cultures exhibit a delay in the appearance of senescence morphology and their life span is extended by approximately 15%. In summary, these data demonstrate the beneficial effect of oleuropein on human fibroblasts undergoing replicative senescence and provide new insights towards enhancement of cellular antioxidant mechanisms by natural compounds that can be easily up-taken through normal diet.

  Induction of growth inhibition and differentiation of human leukemia HL-60 cells by a Tunisian gerboui olive leaf extract.:Biosci Biotechnol Biochem. 2007 May;71(5):1306-12. Epub 2007 May 7. Abaza L, Talorete TP, Yamada P, Kurita Y, Zarrouk M, Isoda H.Laboratory of Characterization and Olive Oil Quality, Biotechnology Center, Hammam Lif, Tunisia.

 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.

  Yellow tea is more potent than other types of tea in suppressing liver toxicity induced by carbon tetrachloride in rats.:Phytother Res. 2007 Jul;21(7):668-70.

 The present study compared the effects of six Chinese teas categorized by their production process: green, white, yellow, oolong, black and pu-erh teas, on carbon tetrachloride (CCl4)-induced liver injury. Wistar rats were given ad libitum the Chinese teas prepared according to the home-style methods for 1 week, and then intraperitoneally injected with CCl4 (1 mg/kg body weight) or olive oil as a vehicle. The yellow tea significantly ameliorated the increase in the activity of the alanine- and aspartate-aminotransferases in plasma. Thus, the drinking of yellow tea may contribute to protection against liver injury.

  Effects of enrichment of refined olive oil with phenolic compounds from olive leaves.:J Agric Food Chem. 2007 May 16;55(10):4139-43. Epub 2007 Apr 17.Paiva-Martins F, Correia R, Félix S, Ferreira P, Gordon MH.Centro de Investiga??o em Química and Departamento de Química, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre, no. 687, 4169-007 Porto, Portugal. mpmartin@fc.up.pt

 The possibility of preparing olive oil, with the same nutritional value and stability characteristics found in virgin olive oil, by the enrichment of refined olive oil with olive leaf polyphenols was studied. To obtain antioxidant phenols similar to those found in virgin olive oil, these components were extracted from the leaves of several olive cultivars from the Northern region of Portugal, namely, Carrasca, Ripa, Negruche, Cordovil, Verdeal, Madural, and Bical cultivars, under several conditions. The concentration of a leaf extract required for addition to refined olive oil to obtain the same stability as virgin olive oil was determined. The extract from 1 kg of leaves was sufficient to fortify 50-320 L of refined olive oil to a similar stability as a virgin olive oil sample depending on the metal concentration of the oil, cultivar, and time of the year when the leaves were picked.

  Colds and influenza: a review of diagnosis and conventional, botanical, and nutritional considerations.:Altern Med Rev. 2007 Mar;12(1):25-48. Review.Roxas M, Jurenka J.Thorne Research, PO Box 25, Dover, ID 83825, USA. m.roxas@comcast.net

 The common cold is the leading cause of doctor visits in the United States and annually results in 189 million lost school days. In the course of one year the U.S. population contracts approximately 1 billion colds. Influenza infection is still a leading cause of morbidity and mortality, accounting for 20-25 million doctor visits and 36,000 deaths per year in the United States. Conventional therapies for colds and flu focus primarily on temporary symptom relief and include over-the-counter antipyretics, anti-inflammatories, and decongestants. Treatment for influenza also includes prescription antiviral agents and vaccines for prevention. This article reviews the common cold and influenza viruses, presents the conventional treatment options, and highlights select botanicals (Echinacea spp., Sambucus nigra, larch arabinogalactan, Astragalus membranaceous, Baptisia tinctoria, Allium sativa, Panax quinquefolium, Eleutherococcus senticosus, Andrographis paniculata, olive leaf extract, and Isatis tinctoria) and nutritional considerations (vitamins A and C, zinc, high lactoferrin whey protein, N-acetylcysteine, and DHEA) that may help in the prevention and treatment of these conditions.


  The effects of polyphenols in olive leaves on platelet function.:Nutr Metab Cardiovasc Dis. 2007 Mar 6;Singh I, Mok M, Christensen AM, Turner AH, Hawley JA.School of Medical Sciences, RMIT University, Melbourne, Australia.

 INTRODUCTION: The phenolic compounds of olive leaves and olive oils in the Mediterranean diet have been associated with a reduced incidence of heart disease. Accordingly, antioxidant-rich diets may prevent the deleterious effects of oxidative metabolism by scavenging free radicals, thus inhibiting oxidation and delaying atherosclerosis. The process involves phospholipase C activation and arachidonic acid metabolism, and is thought to reduce hydrogen peroxide (H(2)O(2)). In our study, an extract of Olea europaea L. leaves was used. The active phenolic compounds in this extract are part of the secoiridoid family, known for their capacity to scavenge H(2)O(2). The results from this study will help to improve our understanding of effects of polyphenol antioxidants in olive leaf extract on platelet function. METHODS: Full blood examination (FBE), platelet aggregation, and ATP release were performed on samples from fasting, normal, healthy male subjects. Platelet function at increasing concentrations of oleuropein was investigated through measures of platelet aggregation and ATP release from activated platelets. RESULTS: Blood analysis (n=11) revealed a significant dose-dependant reduction in platelet activity with olive extract concentrations of 1.0% v/v (P<0.001). ATP Release showed a similar pattern (P=0.02). CONCLUSIONS: Olive leaf polyphenols derived from O. europaea L. leaves inhibited in vitro platelet activation in healthy, non-smoking males. Further bioavailability studies need to be undertaken to determine the in vivo effect of extract on platelet function and to validate the present results.

  Discovery of small-molecule HIV-1 fusion and integrase inhibitors oleuropein and hydroxytyrosol: Part I. fusion [corrected] inhibition.:Biochem Biophys Res Commun. 2007 Mar 23;354(4):872-8. Epub 2007 Jan 24. Erratum in: Biochem Biophys Res Commun. 2007 May 18;356(4):1068.Abaza L, Talorete TP, Yamada P, Kurita Y, Zarrouk M, Isoda H.Laboratory of Characterization and Olive Oil Quality, Biotechnology Center, Hammam Lif, Tunisia.

 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.

  Influence of rosemary (Rosmarinus officinalis, L.) on plant sterol oxidation in extra virgin olive oil.:Mol Nutr Food Res. 2006 Sep;50(9):818-23.D'Evoli L, Huikko L, Lampi AM, Lucarini M, Lombardi-Boccia G, Nicoli S, Piironen V.Department of Applied Chemistry and Microbiology, Food Chemistry, University of Helsinki, Helsinki, Finland. devoli@inran.it

 The objective of this study was to investigate the potential role of rosemary compounds in inhibiting the plant sterol oxidation in extra virgin olive oil during heating. The stability of plant sterols was measured by quantification of plant sterol and sterol oxide formation upon 6 h of heating in both the extra virgin olive oil and its respective oil at 10% rosemary concentration (ROE). The total sterol and sitosterol oxide contents were determined by GC-flame ionization detection (GC-FID) and GC-MS techniques, respectively. Heating experiments were carried out at 180 degrees C for 0, 1, 3 and 6 h. The total sterol content in the extra virgin olive oil was 255 mg/100 g and that in the ROE 270 mg/100 g. Sitosterol was the most abundant sterol in both samples (50% of total sterol). The ROE showed a lower content of sitosterol oxides with respect to the extra virgin olive oil during heating: after 6 h of heating only 6.1% of sitosterol oxides were formed, while up to 11.5% of sitosterol oxidized in the extra virgin olive oil. Our findings suggested that rosemary compounds were able to counteract the oxidation of plant sterols in the extra virgin olive oil during heating, preventing formation of potentially harmful compounds to human health.

  Multivariate optimisation of the microwave-assisted extraction of oleuropein and related biophenols from olive leaves.:Anal Bioanal Chem. 2006 Jun;385(4):753-9. Epub 2006 May 17.Japón-Luján R, Luque-Rodríguez JM, Luque de Castro MD.Analytical Chemistry Department, Annex Marie Curie Building, Rabanales Campus, University of Córdoba, 14071 Córdoba, Spain.

 Microwave assistance is proposed for the first time in order to accelerate the extraction of biophenols from olive leaves. Under optimal working conditions, obtained using a multivariate methodology, complete extraction of the target analytes was achieved in 8 min. The extracts required no clean-up nor concentration prior to injection into a chromatograph-photodiode array detector assembly for individual separation-quantification. The optimal extractant (an 80:20 ethanol-water mixture) was also used in the development of a stirring-based extraction method which required around 24 h for complete extraction of the target compounds. These mixtures can be used as replacements for toxic extractants, with a view to exploiting olive leaves in order to obtain biophenols for human use.

  Novel Secoiridoid glucosides in Olea europaea leaves suffering from boron deficiency.:Biosci Biotechnol Biochem. 2006 Aug;70(8):1898-903.Karioti A, Chatzopoulou A, Bilia AR, Liakopoulos G, Stavrianakou S, Skaltsa H.Department of Pharmacognosy and Chemistry of Natural Products, School of Pharmacy, Athens, Greece.

 From the methanol extract of boron deficient Olea europaea leaves, two secoiridoid glycosides, not detected in leaf extracts of untreated plants, 6'-E-p-coumaroyl-secologanoside and 6'-O-[(2E)-2,6-dimethyl-8-hydroxy-2-octenoyloxy]-secologanoside, were isolated together with three known secoiridoid glycosides, oleuropein, oleoside dimethyl ester, and secologanoside. The structures of the isolated compounds were established by means of NMR and MS spectral analyses. The above novel secoiridoids were synthesized by the plant as a physiological response to nutrient stress.


  Dynamic ultrasound-assisted extraction of oleuropein and related biophenols from olive leaves.:J Chromatogr A. 2006 Mar 3;1108(1):76-82. Epub 2006 Jan 26.Japón-Luján R, Luque-Rodríguez JM, Luque de Castro MD.Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, E-14071 Córdoba, Spain.

 A continuous approach for the ultrasound-assisted extraction of olive biophenols (OBPs) from olive leaves is proposed. Multivariate methodology was used to carry out a detailed optimisation of extraction. Under the optimal working conditions, complete extraction of the target analytes (namely, oleuropein, verbacoside, apigenin-7-glucoside and luteolin-7-glucoside with LODs 11.04, 2.68, 1.49 and 3.91 mg/kg, respectively) was achieved in 25 min. The extract was injected into a chromatograph-photodiode array detector assembly (HPLC-DAD) for individual separation-quantification. No clean-up or preconcentration steps were required. Gas chromatography-mass spectrometry (without derivatization of the analytes) was used to identify OBPs at concentrations below the LODs obtained by HPLC-DAD. The efficacy of ethanol-water mixtures to extract OBPs from olive leaves has been demonstrated and compared with that of a conventional method which requires 24h for complete extraction; so these mixtures can substitute toxic extractants used to date.

  Factors affecting the contents of iridoid oleuropein in olive leaves (Olea europaea L.).:J Agric Food Chem. 2006 Jan 25;54(2):434-40.Ranalli A, Contento S, Lucera L, Di Febo M, Marchegiani D, Di Fonzo V.Istituto Sperimentale per l'Elaiotecnica, Viale Petruzzi 75, 65013 Città S. Angelo, Pescara, Italy. alfonso.ranalli@entecra.it

 In this study, for the first time, the impact of the genetic factor on the contents of oleuropein in olive leaves was not only evaluated but the influence exerted by the color/age of leaves (green, green-yellowish, and yellow) and the collecting period (spring or autumn) was also evaluated. A repetitive high-resolution gas chromatographic quantitation method and an accurate high-performance liquid chromatographic method were developed. These analytical methods gave results showing a highly linear relationship. Samples of olive leaves were taken from seven major Italian olive cultivars, such as Dritta, Leccino, Caroleo, Coratina, Castiglionese, Nebbio, and Grossa di Cassano. Such a vegetal raw material could actually be exploited for recovering oleuropein, considered to be a high-added value molecule. This could be converted into hydrxytyrosol, a compound known to possess strong bioactive properties. Olive leaves showed considerable contents of oleuropein, which with some cultivars were even higher with respect to those present in the corresponding olive fruits (reported in the literature). The amounts of oleuropein in the collected leaves were markedly modified by the color/age and genetic factors, whereas meaningless variations were ascribable to the quantitation method and the collecting period factors. Various chemometrics, applied to the obtained analytical data, appeared to be effective in discriminating the samples on the basis of the above-examined experimental factors, thus confirming how these should be taken into account in future industrial recovery of oleuropein from olive leaves.

  Natural compounds derived from foods modulate nitric oxide production and oxidative status in epithelial lung cells.:J Agric Food Chem. 2005 Dec 28;53(26):9934-9.Zaslaver M, Offer S, Kerem Z, Stark AH, Weller JI, Eliraz A, Madar Z.Institute of Biochemistry, Food Science and Nutrition, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel.

 The effects of natural antioxidants on nitric oxide (NO) modulation and oxidative status were determined in rat epithelial lung cells (L-2). Cells were stimulated with cytokines and treated with one of the following: resveratrol, soybean saponin group B (SSB), quercetin, genistein, olive leaf polyphenol concentrate (OLPC), or N-acetyl-L-cystein (NAC). NAC had no effect on NO levels, whereas resveratrol and OLPC were found to be effective in reducing nitrite levels, modifying iNOS mRNA, and decreasing free radical production. OLPC affected the levels of MnSOD while resveratrol did not, indicating that they act via different pathways. Quercetin and genistein reduced nitrite levels without affecting iNOS levels, presumably by scavenging NO. SSB did not affect nitrite levels, but exposure did reduce iNOS mRNA expression and protein levels, possibly due to antioxidant activity. Naturally occurring antioxidants, in particular resveratrol and OLPC, may have therapeutic potential in the treatment of inflammatory diseases.

  Hypoglycemic and antioxidant effect of oleuropein in alloxan-diabetic rabbits..:Life Sci. 2006 Feb 16;78(12):1371-7. Epub 2005 Oct 19.Al-Azzawie HF, Alhamdani MS.Biochemistry Department, College of Science, University of Baghdad, Baghdad, Iraq.

 Patients with diabetes mellitus are likely to develop certain complication such as retinopathy, nephropathy and neuropathy as a result of oxidative stress and overwhelming free radicals. Treatment of diabetic patients with antioxidant may be of advantage in attenuating these complications. Oleuropein, the active constituent of olive leaf (Olea europaea), has been endowed with many beneficial and health promoting properties mostly linked to its antioxidant activity. This study aimed to evaluate the significance of supplementation of oleuropein in reducing oxidative stress and hyperglycemia in alloxan-induced diabetic rabbits. After induction of diabetes, a significant rise in plasma and erythrocyte malondialdehyde (MDA) and blood glucose as well as alteration in enzymatic and non-enzymatic antioxidants was observed in all diabetic animals. During 16 weeks of treatment of diabetic rabbits with 20 mg/kg body weight of oleuropein the levels of MDA along with blood glucose and most of the enzymatic and non-enzymatic antioxidants were significantly restored to establish values that were not different from normal control rabbits. Untreated diabetic rabbits on the other hand demonstrated persistent alterations in the oxidative stress marker MDA, blood glucose and the antioxidant parameters. These results demonstrate that oleuropein may be of advantage in inhibiting hyperglycemia and oxidative stress induced by diabetes and suggest that administration of oleuropein may be helpful in the prevention of diabetic complications associated with oxidative stress.

  Differential effects of oleuropein, a biophenol from Olea europaea, on anionic and zwiterionic phospholipid model membranes.:Chem Phys Lipids. 2005 Oct;137(1-2):2-17. Epub 2005 Jun 21.Caturla N, Pérez-Fons L, Estepa A, Micol V.Instituto de Biología Molecular y Celular, Universidad Miguel Hernández. Avda. de la Universidad s/n, 03202-Elche, Alicante, Spain.

 Oleuropein (Ole) is the major phenolic constituent of the olive leaf (Olea europaea) and it is also present in olive oil and fruit. In the last years several compounds from olive tree, oleuropein among them, have shown a variety of biological activities such as antimicrobial or antioxidant. A phospholipid model membrane system was used to study whether the Ole biological effects could be membrane related. Ole showed a significant partition level in phospholipid membranes, i.e. 80%, at lipid-saturating conditions. Moreover, fluorescence quenching experiments indicated a shallow location for Ole in membranes. Ole promoted weak effects on zwiterionic phospholipids such as phosphatidylcholine or phosphatidylethanolamine. In contrast, differential scanning microcalorimetry, light scattering and fluorescence anisotropy pH titration studies revealed strong effects on anionic phospholipids such as phosphatidylglycerol at physiological pH and salt conditions. These effects consisted on perturbations at the phospholipid membrane surface, which might involve specific molecular interactions between Ole and the negatively charged phosphate group and therefore modify the phospholipid/water interface properties. It is proposed that Ole induces lipid structures similar to the gel-fluid intermediate phase (IP) described for PG membranes, in a similar way than low ionic strength does. These effects on phosphatidylglycerol may account for the antimicrobial activity of Ole.


  The olive leaf extract exhibits antiviral activity against viral haemorrhagic septicaemia rhabdovirus (VHSV).:Antiviral Res. 2005 Jun;66(2-3):129-36. Epub 2005 Apr 18. Micol V, Caturla N, Pérez-Fons L, Más V, Pérez L, Estepa A.Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, E-03202-Elche, Alicante, Spain.

 A commercial plant extract derived from olive tree leaf (Olea europaea) (LExt) and its major compound, oleuropein (Ole), inhibited the in vitro infectivity of the viral haemorrhagic septicaemia virus (VHSV), a salmonid rhabdovirus. Incubation of virus with LExt or Ole before infection reduced the viral infectivity to 10 and 30%, respectively. Furthermore, LExt drastically decreased VHSV titers and viral protein accumulation (virucidal effect) in a dose dependent manner when added to cell monolayers 36 h post-infection. On the other hand, both the LExt and Ole were able to inhibit cell-to-cell membrane fusion induced by VHSV in uninfected cells, suggesting interactions with viral envelope. Therefore, we propose that O. europaea could be used as a potential source of promising natural antivirals, which have demonstrated to lack impact on health and environment. In addition, Ole could be used to design other related antiviral agents.

  Analysis and quantification of flavonoidic compounds from Portuguese olive (Olea europaea L.) leaf cultivars.:Nat Prod Res. 2005 Feb;19(2):189-95.Meirinhos J, Silva BM, Valent?o P, Seabra RM, Pereira JA, Dias A, Andrade PB, Ferreres F.Escola Superior Agrária, Instituto Politécnico de Bragan?a, Quinta de Sta Apolónia, 5300 Bragan?a, Portugal.

 Twenty three samples of 18 Portuguese olive leaf cultivars were analysed by a reversed-phase HPLC/DAD procedure and eight flavonoidic compounds were identified and quantified (luteolin 7,4'-O-diglucoside, luteolin 7-O-glucoside, rutin, apigenin 7-O-rutinoside, luteolin 4'-O-glucoside, luteolin, apigenin and diosmetin). Luteolin 7,4'-O-diglucoside and luteolin 4'-O-glucoside were identified by HPLC/DAD/MS/MS - ESI. The studied olive leaf samples showed a common phenolic pattern, in which luteolin 4'-O-glucoside was almost always the major compound.

  Boron deficiency and concentrations and composition of phenolic compounds in Olea europaea leaves: a combined growth chamber and field study.:Tree Physiol. 2005 Mar;25(3):307-15.Liakopoulos G, Karabourniotis G.Laboratory of Plant Physiology, Department of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, Botanikos, 11855, Athens, Greece. g_liak@aua.gr

 Boron deficiency is the most frequent micronutrient disorder in olive (Olea spp.) orchards. We tested the hypothesis that plant boron status affects phenolic metabolism, which, in turn, influences several ecophysiological traits of olive (Olea europaea L.) trees, by studying the effects of boron deficiency on leaf phenolic compounds of olive in a growth chamber experiment (CE) and a field experiment (FE). In the CE, a semi-hydroponic system was used to control nutrient supply. Plants received complete nutrient solution containing either 23 (control) or 0 microM H3BO3 (boron-deficient treatment). In the FE, boron-deficient trees were chosen based on visible boron-deficiency symptoms and analysis of their leaf boron concentration. Boron deficiency caused significant accumulation of phenolic compounds in leaves of CE plants (1.7 to 5.8 times more, depending on leaf age), but not in leaves of FE plants. However, in both experiments, the concentration of an unidentified phenolic compound, with a UV-spectrum resembling that of caffeic acid, increased in response to boron deficiency (by a factor of 40 to 184 in the CE and by a factor of three in the FE). Regression analysis showed that the concentration of this compound was negatively correlated to leaf boron concentration, irrespective of growth conditions and treatment. We conclude that, under field conditions, boron deficiency may not be the only factor determining the concentration of total phenolics, but it may be responsible for the accumulation of a distinct phenolic metabolite in olive leaves.

  1,5-Anhydroxylitol from leaves of Olea europaea.:Carbohydr Res. 2004 Nov 15;339(16):2731-2. Campeol E, Flamini G, Cioni PL, Morelli I, D'Andrea F, Cremonini R.Dipartimento di Chimica Bioorganica e Biofarmacia, Università di Pisa, via Bonanno 33, I-56126 Pisa, Italy. camelis@farm.unipi.it

 1,5-Anhydroxylitol, a compound never found previously in the vegetal kingdom was obtained from Olea europaea leaves in approximately 0.5-1% yield.

  Ruminal fermentation and degradation patterns, protozoa population and urinary purine derivatives excretion in goats and wethers fed diets based on olive leaves.:J Anim Sci. 2004 Oct;82(10):3006-14. Ruiz DR, García AI, Moumen A, Alcaide EM.Unidad de Nutrición Animal, Estación Experimental del Zaidín (CSIC), Armilla (Granada), Spain.

 Olives leaves, accrued during the processing of olive harvests for oil extraction, are poor in N, rich in crude fat and ADF (1.19, 8.03 and 28.2 g/100 g of DM, respectively), and relatively low in condensed tannins (11.1 mg/g of DM). Three experiments were conducted in a 2 x 3 (two animal species: goats vs. wethers; and three experimental diets: olive leaves without or with polyethylene glycol supply and olive leaves supplemented with barley and faba beans) factorial design to evaluate ruminal degradation and passage kinetics (Exp. 1), fermentation pattern and protozoa population (Exp. 2), and urinary purine derivatives excretion (Exp. 3). Polyethylene glycol was supplied to evaluate the effects of condensed tannins contained in olive leaves. Ruminal degradability of CP was low in both goats and wethers, although goats showed higher (P < 0.05) values than wethers. Supplementation of olive leaves with barley and faba beans increased (P < 0.001) ruminal degradability of DM and CP. Both goats and wethers fed olive leaves showed similarly low particulate fractional passage rates (0.021 and 0.023/h, respectively). Ingestion of olive leaves promoted low NH3-N and VFA concentrations, which reflect poor microbial activity. These concentrations, especially that of VFA, increased when barley and faba beans were added. Ingestion of olive leaves affected ruminal protozoa: Entodiniomorphida showed low concentrations and Holotricha completely disappeared. When animals received a diet based on olive leaves, barley, and faba beans, Holotricha appeared in the ruminal liquor and Entodiniomorphida increased (P < 0.001). In goats and wethers fed olive leaves alone, urinary allantoin excretion was very low (163 and 164 micromol/kg BW0.75 in goats and wethers, respectively), and moderate values (352 and 389 micromol/kg BW0.75 in goats and wethers, respectively) were observed when a diet of olive leaves, barley, and faba beans was fed. The polyethylene glycol supply did not have an effect in goats or in wethers, indicating the lack of an effect of condensed tannins in olive leaves. Ingestion of olive leaves promotes a low microbial activity, although its supplementation with readily degraded carbohydrates and protein improves microbial activity and, as a consequence, increases its ruminal degradation. In general, for most of the measured variables, there were no animal species x diet interactions. Thus, goats and wethers had similar ruminal activities when fed diets based on olive leaves.


  Countercurrent supercritical fluid extraction and fractionation of high-added-value compounds from a hexane extract of olive leaves.:J Agric Food Chem. 2004 Jul 28;52(15):4774-9.Tabera J, Guinda A, Ruiz-Rodríguez A, Se?oráns FJ, Ibá?ez E, Albi T, Reglero G.Area de Tecnología de Alimentos, Facultad de Ciencias, Universidad Autónoma de Madrid, 28049 Cantoblanco, Madrid, Spain. javier.tabera@uam.es

 Countercurrent supercritical fluid extraction (CC-SFE) at a pilot scale plant was used for fractionation of high-added-value products from a raw extract of olive leaves in hexane. Compounds found in the raw extract were waxes, hydrocarbons, squalene, beta-carotene, triglycerides, alpha-tocopherol, beta-sitosterol, and alcohols. The CC-SFE extraction process was investigated according to a 2(3) full factorial experimental design using the following variables and ranges: extraction pressure, 75-200 bar; extraction temperature, 35-50 degrees C; and ethanol as modifier, 0-10%. Data were analyzed in terms of extraction yield, enrichment, recovery, and selectivity. Higher extraction yields were attained at 200 bar. For most of the compounds analyzed enrichment was attained at the same conditions, that is, 75 bar, 35 degrees C, and 10% ethanol. Hydrocarbons were usually recovered in the separators, whereas waxes and alpha-tocopherol remain in the raffinate. Selectivity data reveal that alpha-tocopherol is the most easily separable compound. The influence of the experimental factors on the recovery of all the compounds was studied by means of regression models. The best fitted model was attained for beta-sitosterol, with R2 = 99.25%.

  Production of highly purified hydroxytyrosol from Olea europaea leaf extract biotransformed by hyperthermophilic beta-glycosidase.:Briante R, Patumi M, Febbraio F, Nucci R.Istituto di Biochimica delle Proteine del CNR, Via Marconi 10-80125 Napoli, Italy.

 A large amount of highly purified hydroxytyrosol (91-94% in weight) is obtained in short time by a simple biotransformation of Olea europaea leaf extract by a partially purified hyperthermophilic beta-glycosidase immobilized on chitosan support. The biotransformation conditions have been modulated for increasing the hydroxytyrosol yield, whilst chitosan and chitin matrices are used as adsorbent materials in liquid phase hydroxytyrosol extraction from the biotransformed mixtures. Natural and non-toxic hydroxytyrosol has been by this way produced from a vegetal source, and this compound appeared for the first time highly purified by natural and biocompatible safe biopolymers in comparison to previous results. Moreover, the GC analyses have displayed that the eluates from a two-step bioreactor have qualitative composition very similar to that of the extra-virgin olive oil polar fraction. The proposed bioreactor could also find application in the utilization of olive mill waste waters (OMWW), medium rich in large amounts of oleuropein, which can be converted in pharmacologically active compounds.

  A gas chromatographic determination of residues of eleven insecticides and two metabolites on olive tree leaves.:J AOAC Int. 2004 Jan-Feb;87(1):146-50.Aplada-Sarlis P, Miliadis GE, Liapis K, Tsiropoulos NG.Benaki Phytopathological Institute, Pesticide Residues Laboratory, 7 Ekalis St, 145 61, Kifissia, Greece.

 A gas chromatographic (GC) method was developed and statistically validated for the simultaneous determination of residues of pyrethroid, endosulfan, and organophosphorus insecticides and some of their metabolites on olive tree leaves. Pesticide residues were extracted by static extraction with acetone-dichloromethane. After evaporation of the extract to dryness and redissolution in acetone, the organophosphorus insecticides were determined by GC with nitrogen-phosphorus detection. Another portion of the extract, after solvent change to acetonitrile, was cleaned up on an Alumina-N cartridge and analyzed for insecticides sensitive to electron-capture detection (ECD), i.e., pyrethroids and endosulfan and its metabolite. Recoveries of the organophosphorus insecticides ranged from 80.7 to 93.3% with relative standard deviations (RSDs) of < or = 7.2%; recoveries of the ECD-sensitive insecticides ranged from 71.6 to 89.5% with RSDs of < or = 11.6%. The method was used to analyze 26 samples of olive tree leaves from organic olive groves all over Greece, and the results confirmed the viability of the method for routine analysis. Residues of fenthion and fenthion sulfoxide were found in one and 3 samples, respectively, and their identities were confirmed by GC with mass spectrometry.

  Anti-HIV activity of olive leaf extract (OLE) and modulation of host cell gene expression by HIV-1 infection and OLE treatment.:Biochem Biophys Res Commun. 2003 Aug 8;307(4):1029-37.Lee-Huang S, Zhang L, Huang PL, Chang YT, Huang PL.Department of Biochemistry, New York University School of Medicine, New York, NY 10016, USA. Sylvia.lee-huang@med.nyu.edu

 We investigated the antiviral activity of olive leaf extract (OLE) preparations standardized by liquid chromatography-coupled mass spectrometry (LC-MS) against HIV-1 infection and replication. We find that OLE inhibits acute infection and cell-to-cell transmission of HIV-1 as assayed by syncytia formation using uninfected MT2 cells co-cultured with HIV-1-infected H9 T lymphocytes. OLE also inhibits HIV-1 replication as assayed by p24 expression in infected H9 cells. These anti-HIV effects of OLE are dose dependent, with EC(50)s of around 0.2 microg/ml. In the effective dose range, no cytotoxicity on uninfected target cells was detected. The therapeutic index of OLE is above 5000. To identify viral and host targets for OLE, we characterized gene expression profiles associated with HIV-1 infection and OLE treatment using cDNA microarrays. HIV-1 infection modulates the expression patterns of cellular genes involved in apoptosis, stress, cytokine, protein kinase C, and hedgehog signaling. HIV-1 infection up-regulates the expression of the heat-shock proteins hsp27 and hsp90, the DNA damage inducible transcript 1 gadd45, the p53-binding protein mdm2, and the hedgehog signal protein patched 1, while it down-regulates the expression of the anti-apoptotic BCL2-associated X protein Bax. Treatment with OLE reverses many of these HIV-1 infection-associated changes. Treatment of HIV-1-infected cells with OLE also up-regulates the expression of the apoptosis inhibitor proteins IAP1 and 2, as well as the calcium and protein kinase C pathway signaling molecules IL-2, IL-2Ralpha, and ornithine decarboxylase ODC1.

  In vitro antimicrobial activity of olive leaves.:Mycoses. 2003 Apr;46(3-4):132-6.Markin D, Duek L, Berdicevsky I.Department of Microbiology, Rappaport Faculty of Medicine, Technion-Institute of Technology, Haifa, Israel.

 We investigated the antimicrobial effect of olive leaves against bacteria and fungi. The microorganisms tested were inoculated in various concentrations of olive leaf water extract. Olive leaf 0.6% (w/v) water extract killed almost all bacteria tested, within 3 h. Dermatophytes were inhibited by 1.25% (w/v) plant extract following a 3-day exposure whereas Candida albicans was killed following a 24 h incubation in the presence of 15% (w/v) plant extract. Olive leaf extract fractions, obtained by dialysis, that showed antimicrobial activity consisted of particles smaller than 1000 molecular rate cutoffs. Scanning electron microscopic observations of C. albicans, exposed to 40% (w/v) olive leaf extract, showed invaginated and amorphous cells. Escherichia coli cells, subjected to a similar treatment but exposed to only 0.6% (w/v) olive leaf extract showed complete destruction. These findings suggest an antimicrobial potential for olive leaves.


  Safety evaluation of olive phenolic compounds as natural antioxidants.:Int J Food Sci Nutr. 2003 May;54(3):159-74.Farag RS, El-Baroty GS, Basuny AM.Department of Biochemistry, Faculty of Agriculture, Cairo University, Giza, Egypt.

 Free and total polyphenolic compounds were extracted from the fruits and leaves of the Picual cultivar. The safety limits of these compounds were recognized by measuring the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the levels of high-density lipoprotein (HDL) cholesterol and total lipids of rat serum. The free and total phenolic compounds (400, 800, and 1600 ppm) and butylated hydroxy toluene (BHT) (200 ppm) were daily ingested for 7 weeks. The administration of olive total and free phenolic compounds at 400 and 800 ppm did not cause any significant changes on ALT and AST activities and serum total lipids. These compounds at 1600 ppm caused significant increase in ALT and AST activities and the content of total lipids. Both olive phenolic compounds were superior to that of BHT in increasing HDL-cholesterol level. Nutritional experiments demonstrated that BHT at 200 ppm caused an enlargement in the kidney and liver of the rat compared with the administration of total and free olive phenolic compounds at 1200 and 1600 ppm. Microscopical examination of kidney and liver tissues of rats administered free and total phenolic compounds at 1200 ppm had the same histological character as that of control rats, while the administration of BHT (200 ppm) and phenolic compounds (1600 ppm) induced severe damage to the tissues of the rat kidney and liver.

  Quantitative changes in phenolic content during physiological development of the olive (Olea europaea) cultivar Hardy's Mammoth.:J Agric Food Chem. 2003 Apr 23;51(9):2532-8. Ryan D, Prenzler PD, Lavee S, Antolovich M, Robards K.School of Science and Technology and Farrer Centre, Charles Sturt University, Locked Bag 588, Wagga Wagga 2678, Australia.

 This investigation was designed to characterize phenolic metabolism of the olive cultivar, Hardy's Mammoth, by examining its constitutive tissues. The phenolic profiles of pulp, seed, stone, and new and old season leaves were monitored over two fruiting seasons, to investigate possible relationships between tissues and phenol content and to determine the impact of alternate fruit bearing. No major qualitative differences in phenolic composition were found between the various tissues; however, distinct differences between the tissues with respect to quantifiable phenols were established. Relationships between 2-(3,4-dihydroxyphenyl)ethyl (3E,4E)-4-formyl-3-(2-oxoethyl)hex-4-enoate ester, oleuropein, and hydroxytyrosol in pulp and leaf were identified and found to be related to alternate bearing. Concentrations of 5-caffeoylquinic acid in old season leaves differed dramatically between seasons, confirming earlier studies.

  Antihypertensive, antiatherosclerotic and antioxidant activity of triterpenoids isolated from Olea europaea, subspecies africana leaves.:J Ethnopharmacol. 2003 Feb;84(2-3):299-305.Somova LI, Shode FO, Ramnanan P, Nadar A.Department of Human Physiology, University of Durban-Westville, Private Bag X54001, Durban 4000, South Africa. somova@pixie.udw.ac.za

 For the first time a biossay-directed study of triterpenoids isolated from the leaves of Olea europaea from Greece, from wild African olive and from a cultivar of O. europaea grown in Cape Town was reported. The experiment was undertaken since our preliminary analyses showed that the African wild olive leave is rich in triterpenoids and contain only traces of the glycoside oleuropein, which is typical for the European olive leaves. The isolate of the African wild olive leaves (AO) used in the experiments was found to contain 0.27% 1:1 mixture of oleanolic acid and ursolic acid, named oleuafricein. The isolate of Greek olive leaves (GO) was found to contain 0.71% oleanolic acid, and the Cape Town cultivar (CT) contained 2.47% oleanolic acid. No ursolic acid was found in either GO or CT. The antihypertensive, diuretic, antiatherosclerotic, antioxidant and hypoglycemic effects of authentic oleanolic and ursolic acid and the three isolates (GO, AO and CT) were studied on Dahl salt-sensitive (DSS), insulin-resistant rat genetic model of hypertension. All three isolates, in a dose 60 mg/kg b.w. for 6 weeks treatment, prevented the development of severe hypertension and atherosclerosis and improved the insulin resistance of the experimental animals. GO, OA and CT isolates could provide an effective and cheap treatment of this particular, most common type of salt-sensitive hypertension in the African population.

  Volatile fractions from three cultivars of Olea europaea L. collected in two different seasons.:J Agric Food Chem. 2003 Mar 26;51(7):1994-9.Campeol E, Flamini G, Cioni PL, Morelli I, Cremonini R, Ceccarini L.Dipartimento di Chimica Bioorganica e Biofarmacia, Università di Pisa, via Bonanno 33, 56126 Pisa, Italy. camelis@farm.unipi.it

 The chemical composition of the volatile fractions from leaves of three Olea europaea L. cultivars (Leccino, Frantoio, and Cipressino) harvested at two different times of the year were examined by GC and GC-MS. The results showed a high content of aliphatic aldehydes in the three cultivars during both harvesting periods and an increase of (E)-2-hexenal (an aldehyde with high antimicrobial properties) percentage from July to November.

  Radioprotective effects in vivo of phenolics extracted from Olea europaea L. leaves against X-ray-induced chromosomal damage: comparative study versus several flavonoids and sulfur-containing compounds.:J Med Food. 2002 Fall;5(3):125-35.Benavente-García O, Castillo J, Lorente J, Alcaraz M.Research and Development Department, Furfural Espa?ol S.A., Camino Viejo de Pliego s/n, 80320 Alcantarilla, Murcia, Spain. laboratorio@furesa.es

 The radioprotective effects of a polyphenolic extract of Olea europaea L. leaves (OL); the flavonoids diosmin and rutin, which are widely used as pharmaceuticals; and the sulfur-containing compounds dimethylsulfoxide (DMSO) and 6-n-propyl-2-thiouracil (PTU) were determined by using the micronucleus test for anticlastogenic activity, evaluating the reduction of the frequency of micronucleated polychromatic erythrocytes (MnPCEs) in bone marrow of mouse before and after X-ray irradiation. With treatment before X-irradiation, the most effective compounds were, in order, rutin > DMSO > OL > PTU > diosmin. These results showed, for the polyphenols studied, a linear correlation (r(2) = 0.965) between anticlastogenic activity and antioxidant capacity. The magnitude of protection with treatment after X-irradiation were lower, and the most effective compounds were, in order, OL > diosmin > rutin; DMSO and PTU lacked radioprotective activity. Therefore, OL is the only substance that showed a significant anticlastogenic activity both before and after X-ray irradiation treatments. Structurally, the free oxygen radicals and lipoperoxyradicals scavenging capacity and, consequently, the anticlastogenic activity of these polyphenolic compounds are based principally on the presence of specific functional groups, mainly catechol groups (rutin, oleuropein, hydroxytyrosol, verbascoside, luteolin), that also increase the stability of the aroxyl-polyphenol radical generated in the above processes.


  Blood pressure lowering effect of an olive leaf extract (Olea europaea) in L-NAME induced hypertension in rats.:Arzneimittelforschung. 2002;52(11):797-802.Khayyal MT, el-Ghazaly MA, Abdallah DM, Nassar NN, Okpanyi SN, Kreuter MH.Department of Pharmacology, Faculty of Pharmacy, Cairo University, Cairo, Egypt. mtkhayyal@hotmail.com

 A specially prepared olive leaf extract (EFLA 943) has been tested for its blood pressure lowering activity in rats rendered hypertensive by daily oral doses of L-NAME (NG-nitro-L-arginine methyl ester, 50 mg/kg) for at least 4 weeks. Oral administration of the extract at different dose levels at the same time as L-NAME for a period of 8 weeks showed a dose dependent prophylactic effect against the rise in blood pressure induced by L-NAME, best effects being induced by a dose of 100 mg/kg of the extract. In rats previously rendered hypertensive by L-NAME for 6 weeks and then treated with that dose of the extract for a further 6 weeks without discontinuation of L-NAME, normalisation of the blood pressure was observed. The findings confirm previous reports on the hypotensive effects of olive leaf. The special extract, EFLA 943, was shown to give consistent results with little individual variability. The antihypertensive effect of the extract may be related to a variety of factors involving reversal of vascular changes involved in the L-NAME induced hypertension.

  Olea europaea L. leaf extract and derivatives: antioxidant properties.:J Agric Food Chem. 2002 Aug 14;50(17):4934-40.Briante R, Patumi M, Terenziani S, Bismuto E, Febbraio F, Nucci R.Istituto di Biochimica delle Proteine ed Enzimologia del CNR, Via Marconi 10, 80125 Napoli, Italy.

 This paper reports a very simple and fast method to collect eluates with high amounts of hydroxytyrosol, biotransforming Olea europaea L. leaf extract by a thermophilic beta-glycosidase immobilized on chitosan. Some phenolic compounds in the leaf tissue and in the eluates obtained by biotransformation are identified. To propose the eluates as natural substances from a vegetal source, their antioxidant properties have been compared with those of the leaf extract from which they are originated. The eluates possess a higher concentration of simple phenols, characterized by a stronger antioxidant capacity, than those available in extra virgin olive oils and in many tablets of olive leaf extracts, commercially found as dietetic products and food integrators.

  Effect of freeze dried extract of Olea europaea on the pituitary-thyroid axis in rats.:Phytother Res. 2002 May;16(3):286-7.Al-Qarawi AA, Al-Damegh MA, ElMougy SA.Department of Veterinary Medicine, College of Agriculture and Veterinary Medicine, King Saud University, Saudi Arabia.

 The effect of an aqueous extract of olive (Olea europaea) leaf on the thyroid activity was studied. The results suggest a stimulatory action of the extract on the thyroid, unrelated to the pituitary.

  Isolation and characterization of the antioxidant component 3,4-dihydroxyphenylethyl 4-formyl-3-formylmethyl-4-hexenoate from olive (Olea europaea) leaves.:J Agric Food Chem. 2001 Sep;49(9):4214-9. Paiva-Martins F, Gordon MH.Centro de Investiga??o em Química, Departamento de Química, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre, number 687, 4169-007 Porto, Portugal.

 Storage of olive (Olea europaea) leaves for 22 h at 37 degrees C in closed plastic bags caused the content of a nonglycosidic secoiridoid, 3,4-dihydroxyphenylethyl 4-formyl-3-formylmethyl-4-hexenoate (3,4-DHPEA-EDA) to rise from 15% to 50% of the phenolic extract with corresponding falls in the content of oleuropein and two oleuropeindials, which were identified as precursors of 3,4-DHPEA-EDA. Pure product was isolated from one set of stored olive leaves in a 0.16% yield. Storage of olive leaves under various conditions showed that the moisture present in closed plastic bags was important for the formation of 3,4-DHPEA-EDA. The time taken to reach the maximum concentration of the product varied widely for different samples of olive leaves, with a shorter time for the sample with lower initial oleuropein content. The oleuropeindial precursors of the product were readily hydrolyzed to carboxylic acid derivatives, which have been identified by NMR. The antiradical activity of 3,4-DHPEA-EDA, evaluated by scavenging of 2,2-diphenyl-1-picrylhydrazyl radicals, was comparable to that of alpha-tocopherol.

  In vitro anti-complementary activity of flavonoids from olive (Olea europaea L.) leaves.:Pharmazie. 1996 Oct;51(10):765-8. Pieroni A, Heimler D, Pieters L, van Poel B, Vlietinck AJ.Department of Pharmaceutical Sciences, University of Antwerp, Belgium.

 From extracts of olive (Olea europaea L., Oleaceae) leaves showing anti-complementary activity, the flavonoids apigenin, apigenin-4'-O-rhamnosylglucoside, apigenin-7-O-glucoside, luteolin, luteolin-4'-O-glucoside, luteolin-7-O-glucoside, chrysoeriol, chrysoeriol-7-O-glucoside and quercetin-3-O-rhamnoside were isolated. Major isolated constituents strongly inhibited the classical pathway of the complement system.


  Vasodilator effect of olive leaf.:Planta Med. 1991 Oct;57(5):417-9.Zarzuelo A, Duarte J, Jiménez J, González M, Utrilla MP.Department of Pharmacology, School of Pharmacy, University of Granada, Spain

 We studied the importance of the smooth vascular muscle endothelium in the vasodilator action of the decoction of olive (Olea europaea) leaf. The decoction caused relaxation of isolated rat aorta preparations both in the presence (IC50 1.12 +/- 0.33 mg/ml) and in the absence (IC50 1.67 +/- 0.16 mg/ml) of endothelium. The results indicate that the relaxant activity of the lyophilized decoction is independent of the integrity of the vascular endothelium. We also showed that oleuropeoside is a component responsible for vasodilator activity but, from the results, it seems likely that at least one other principle is to be found in the olive leaf which is either a vasodilator itself or else potentiates the relaxant effect of oleuropeoside.

  Peroxidases and ethylene formation in olive tree leaves.:Rev Esp Fisiol. 1989 Mar;45(1):47-52. Spanish.Vioque B, Fernández-Maculet JC, Albi MA, Castellano JM, Vioque A.Departamento de Fisiología y Tecnología Postrecolección, Instituto de la Grasa y sus Derivados, Sevilla, Espa?a.

 The loss of intercellular peroxidases from olive tree leaf sections subjected to isoelectric focusing correlates well with the capacity of the referred sections to convert 1-aminocyclopropane-1-carboxylic acid (ACC) into ethylene. Both activities are compared in leaves of different ages (according to their position on the branch) and it is found that they follow a parallel course. The inhibition of the capacity to convert ACC into ethylene in homogenized olive tree leaves is due to inhibitors release. The results, in relation to other authors' reports, are discussed.

Reference:

• 1.Olive leaf was first used medicinally in Ancient Egypt.

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♣ last edit date:Nov 19th 2012 at 14:30