Antiprotozoal Activity of Turkish Origanum onites Essential Oil and Its Components.

Essential oil of Origanum species is well known for antimicrobial activity, but only a few have been evaluated in narrow spectrum antiprotozoal assays. Herein, we assessed the antiprotozoal potential of Turkish Origanum onites L. oil and its major constituents against a panel of parasitic protozoa. The essential oil was obtained by hydrodistillation from the dried herbal parts of O. onites and analyzed by Gas Chromatography-Flame Ionization Detector (GC-FID) and Gas Chromatography coupled with Mass Spectrometry (GC-MS). The in vitro activity of the oil and its major components were evaluated against Trypanosoma brucei rhodesiense, T. cruzi, Leishmania donovani, and Plasmodium falciparum. The main component of the oil was identified as carvacrol (70.6%), followed by linalool (9.7%), p-cymene (7%), γ-terpinene (2.1%), and thymol (1.8%). The oil showed significant in vitro activity against T. b. rhodesiense (IC50 180 ng/mL), and moderate antileishmanial and antiplasmodial effects, without toxicity to mammalian cells. Carvacrol, thymol, and 10 additional abundant oil constituents were tested against the same panel; carvacrol and thymol retained the oil's in vitro antiparasitic potency. In the T. b. brucei mouse model, thymol, but not carvacrol, extended the mean survival of animals. This study indicates the potential of the essential oil of O. onites and its constituents in the treatment of protozoal infections.

Repellency of the Origanum onites L. essential oil and constituents to the lone star tick and yellow fever mosquito.

The oregano, Origanum onites L., essential oil (EO) was tested in laboratory behavioural bioassays for repellent activity against Amblyomma americanum (L.) and Aedes aegypti (L.). The O. onites EO was characterised using GC-FID and GC-MS. Carvacrol (75.70%), linalool (9.0%), p-cymene (4.33%) and thymol (1.9%) were the most abundant compounds. At a concentration of 0.413 mg oil/cm2 of filter paper, O. onites EO repelled 100% of the ticks tested and at 0.103 mg oil/cm2 of filter paper, 66.7% of the ticks were repelled. At 0.075 mg oil/cm2 filter paper, thymol repelled 66.7% of the ticks compared to 28.7% by carvacrol at that same concentration. Against Ae. aegypti, O. onites EO was repellent at the minimum effective dosage (MED) of 0.011 (±0.00) mg/cm2 in the cloth patch assay compared to the reference control, N,N-dimethyl-3-methylbenzamide (DEET) with a MED = 0.007 ± (0.003) mg/cm2.

Inhibition of Human Neutrophil Responses by the Essential Oil of Artemisia kotuchovii and Its Constituents.

Essential oils were obtained by hydrodistillation of the flowers+leaves and stems of Artemisia kotuchovii Kupr. (AKEO(f+l) and AKEO(stm), respectively) and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The primary components of the oils were estragole, (E)- and (Z)-ß-ocimenes, methyleugenol, limonene, spathulenol, ß-pinene, myrcene, and (E)-methyl cinnamate. Seventy-four constituents were present at concentrations from 0.1 to 1.0%, and 34 compounds were identified in trace (<0.1%) amounts in one or both plant components. Screening of the essential oils for biological activity showed that AKEO(stm), but not AKEOf+l, inhibited N-formyl-Met-Leu-Phe (fMLF)-stimulated Ca(2+) flux and chemotaxis and phorbol-12-myristate-13-acetate (PMA)-induced reactive oxygen species (ROS) production in human neutrophils. Selected pure constituents, representing >96% of the AKEO(stm) composition, were also tested in human neutrophils and HL-60 cells transfected with N-formyl peptide receptor 1 (FPR1). One component, 6-methyl-3,5-heptadien-2-one (MHDO), inhibited fMLF- and interleukin 8 (IL-8)-stimulated Ca(2+) flux, fMLF-induced chemotaxis, and PMA-induced ROS production in human neutrophils. MHDO also inhibited fMLF-induced Ca(2+) flux in FPR1-HL60 cells. These results suggest that MHDO may be effective in modulating some innate immune responses, possibly by inhibition of neutrophil migration and ROS production.

In vitro antioxidant properties and anthocyanin compositions of elderberry extracts.

In this study, dried elderberry fruits growing wild in Turkey were macerated using different solvents and an infusion was prepared according to traditional methods. All extracts were investigated for their total phenolic content, total monomeric anthocyanins, qualitative-quantitative determination of cyanidin-3-glucoside (by HPLC-UV analysis), anthocyanin compositions (by LC/MS-MS), free radical scavenging activity (DPPH and ABTS) and inhibition of ß-carotene/linoleic acid co-oxidation. An extract with 70% ethanol was found to be richer in cyanidin-3-glucoside when compared to the other extracts. The infusion was found to be as rich as the 70% ethanol extract. Ethanol and acetone extracts (both 70%) were found to be more active in the free radical activity and ß-carotene bleaching assays. Water extract showed good ABTS radical scavenging activity when compared with ascorbic acid.

Phoenix dactylifera L. spathe essential oil: chemical composition and repellent activity against the yellow fever mosquito.

Date palm, Phoenix dactylifera L. (Arecaceae), grows commonly in the Arabian Peninsula and is traditionally used to treat various diseases. The aim of the present study was to identify chemical composition of the essential oil and to investigate the repellent activity. The essential oil of P. dactylifera was obtained by hydrodistillation from the spathe, a specialized leaf structure that surrounds the pollinating organs of the palm. The oil was subsequently analyzed by GC-FID and GC-MS. The oil showed promising repellent activity against yellow fever mosquito - Aedes aegypti. Sixteen components were characterized, constituting 99% of the oil. The main components were 3,4-dimethoxytoluene (73.5%), 2,4-dimethoxytoluene (9.5%), ß-caryophyllene (5.5%), p-cresyl methyl ether (3.8%), and caryophyllene oxide (2.4%). The minimum effective dosage (MED) for repellency for the P. dactylifera oil was 0.051mg/cm(2), which had moderately lower potency compared to reference standard N,N-diethyl-3-methylbenzamide, DEET (0.018mg/cm(2)) in the "cloth patch assay". The five major compounds were individually assayed for repellency to determine to what extent each is responsible for repellency from the oil. 3,4-Dimethoxytoluene and 2,4-dimethoxytoluene showed the best repellent activity with the same MED value of 0.063mg/cm(2), respectively. The results indicate that these two constituents which comprise a large proportion of the P. dactylifera oil (83%) are likely responsible for the observed repellent activity. In this aspect, the P. dactylifera spathe oil is a sustainable, promising new source of natural repellents.

Anticandidal activity of the essential oil of Nepeta transcaucasica Grossh.

Hydrodistallation of the aerial parts of Nepeta transcaucasica Grossh. (Lamiaceae), collected in Agri, Dogubayazit Province, afforded an essential oil that was characterized by GC and GC/MS analyses. Twenty-seven compounds, representing 97.69% of the total oil composition, were identified, and 4aα,7α,7aß-nepetalactone (1; 39%), 4aα,7α,7aα-nepetalactone (2; 28%), and germacrene D (3; 15%) constituted the major components. The anticandidal effects of the oil were evaluated against seven Candida strains by using the broth microdilution method. The oil showed good inhibitory effects against C. glabrata and C. tropicalis at minimal inhibitory concentrations (MICs) of 0.09 and 0.375 mg/ml, respectively.

Iron(III) reducing and antiradical activities of three Sideritis from Turkey.

CONTEXT: Sideritis species (Lamiaceae) are widely used as herbal tea and have been used in folk medicine for their anti-inflammatory, anti-rheumatic, digestive, and antimicrobial activities in Turkey. Sideritis dichotoma Huter., Sideritis erythrantha Boiss. var. cedrotorum, and Sideritis vuralii H. Duman et Baser are available as commercial products in Turkey. OBJECTIVE: The antiradical activities of the various solvent extracts of Sideritis species are investigated here for the first time. MATERIALS AND METHODS: Plant samples were sequentially extracted with n-hexane, dichloromethane, methanol, and aqueous methanol (50%, v/v) in Soxhlet apparatus. The extracts of Camellia sinensis (L.) Kuntze (Theaceae) were also prepared for use as a positive control. Total phenolics, iron(III) reductive effects, and 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging activities of the all extracts were measured colorimetrically. RESULTS: The aqueous MeOH and MeOH extracts contained the highest amount of total phenols, whereas the n-hexane extract contained the lowest amounts. The polar extracts of C. sinensis showed higher antiradical activity and also iron(III) reductive effects than the Sideritis species; however, the non-polar extracts of Sideritis species were found to be more active than those from C. sinensis in the iron(III) reductive assay and in the DPPH(•) assay as well. But none of the extracts was found to be as active as with positive controls, viz., ascorbic acid, butylated hydroxyanisole (BHA), and Trolox. DISCUSSION AND CONCLUSION: These results can be shown to have antioxidant activities of these Sideritis species and support the ethnopharmacological use of these Sideritis plants.

Essential oil compositions and antioxidant properties of the roots of twelve Anatolian Paeonia taxa with special reference to chromosome counts.

Essential oil compositions and antioxidant potentials of fourteen ethanol (75%) root extracts prepared from twelve taxa of the genus Paeonia (Paeoniaceae), including P. arietina Anders., P. daurica Andrews, P. xkayae N. Ozhatay, P. kesrouanensis Thiéb., P. mascula (L.) Miller subsp. arasicola G. Kaynak, ö. Yilmaz & R. Daskin, P. mascula (L.) Miller subsp. bodurii N. Ozhatay, P. cf. mascula L. (Mill.) subsp. mascula (two samples from central and northeastern Anatolia), P. cf. officinalis Retz., P. peregrina Miller (two samples from western and northwestern Anatolia), P. tenuifolia L., P. turcica Davis & Cullen, and P. wittmanniana Hartwiss ex Lindl. were assessed. The chromosome numbers of the root tips of the species were examined using chromosome staining technique with Shiff's reagent under Leitz microscope. The essential oils of the roots of the Paeonia species were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) and the major components were identified as salicylaldehyde (10%-94.4%), cis-myrtanal (5.5%-59.7%), and methyl salicylate (2%-52.2%). Antioxidant potentials were tested against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide (NO) radicals using propyl gallate and rutin as the references. Total phenolic contents of the ethanol extracts were determined using Folin-Ciocalteau's method. The extracts exerted moderate NO scavenger effect and displayed insignificant DPPH radical scavenger activity at 500 microg mL(-1). On the other hand, P. daurica, P. tenuifolia and P. cf. mascula subsp. mascula are diploids with 2n = 10, while other nine taxa are tetraploids with 2n = 20.

Biological and pharmacological activities of carvacrol and carvacrol bearing essential oils.

Oregano essential oils obtained from the genera Origanum, Thymus, Coridothymus, Thymbra, Satureja and Lippia are rich in carvacrol, a monoterpenic phenol isomeric with thymol. Turkey is the biggest exporter of oregano herb and oil to the world markets. Oregano is mainly used in food, spice and pharmaceutical industries. Carvacrol is responsible for the biological activities of oregano. Many diverse activities of carvacrol such as antimicrobial, antitumor, antimutagenic, antigenotoxic, analgesic, antispasmodic, antiinflammatory, angiogenic, antiparasitic, antiplatelet, AChe inhibitory, antielastase, insecticidal, antihepatotoxic and hepatoprotective activities and uses such as feed additive, in honeybee breeding and in gastrointestinal ailments have been shown. This paper highlights these activities and attempts to explain the possible in vivo mechanism of action of carvacrol.

Effect of maturation on the composition and biological activity of the essential oil of a commercially important Satureja species from Turkey: Satureja cuneifolia Ten. (Lamiaceae).

The essential oil obtained by hydrodistillation from aerial parts of Satureja cuneifolia Ten., collected in three different maturation stages such as preflowering, flowering, and postflowering, were analyzed simultaneously by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Thymol (42.5-45.2%), p-cymene (19.4-24.3%), and carvacrol (8.5-13.2%) were identified as the main constituent in all stages. At the same time, the essential oils and main components were evaluated for their antimicrobial activity using a microdilution assay resulting in the inhibition of a number of common human pathogenic bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and the yeasts Candida albicans and Candida tropicalis. The minimum inhibitory concentrations (MIC) varied between 62.5 and 250 microg/mL within a moderate antimicrobial activity range. Furthermore, the antioxidant capacity of the essential oils and major components thymol and carvacrol were examined in vitro. The essential oils obtained from S. cuneifolia in three different stages and its main components were interacted with 1,1-diphenyl-2-picrylhydrazyl (DPPH (*)) as a nitrogen-centered stable radical, resulting in IC 50 = 1.6-2.1 mg/mL. In addition, the effects on inhibition of lipid peroxidation of the essential oils were assayed using the beta-carotene bleaching method. All of the tested oils inhibited the linoleic acid peroxidation at almost the same level as butylated hydroxytoluene (BHT) (93.54-94.65%). BHT and ascorbic acid were used as positive controls in the antioxidant assays.

The effects of intra-rectal and intra-peritoneal application of Origanum onites L. essential oil on 2,4,6-trinitrobenzenesulfonic acid-induced colitis in the rat.

The aim of the present study is to investigate the treatment efficiency of intra-rectal (IR) and intra-peritoneal (IP) application of Origanum onites essential oil (OOEO), which is a well-known antioxidant, in the colitis model induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) and ethanol (E) in comparison with dexamethasone therapy through the morphologic damage score. Monoclonal antibodies against intercellular adhesion molecule-1 (ICAM-1, CD54), anti-rat granulocytes, and myeloperoxidase (MPO), were also investigated immunohistochemically. There was a significant difference in terms of ulceration, mucus cell depletion, inflammatory cell infiltration, vascular dilatation (p<0.001), crypt abscesses (p<0.01), and edema (p<0.05) between OOEO-1mg/kg-IR and control colitis groups. A significant difference was encountered in terms of mucus cell depletion, crypt abscesses, inflammatory cell infiltration, vascular dilatation (p<0.01), and ulceration (p<0.05) between the OOEO-0.1mg/kg-IR and control colitis groups. A significant difference was noticed in terms of ulceration, inflammatory cell infiltration, mucus cell depletion (p<0.001), vascular dilatation (p<0.01), and mucosal atrophy (p<0.05) between the OOEO-1mg/kg-IP and control colitis groups. There was a significant difference in terms of ulceration, mucus cell depletion, inflammatory cell infiltration (p<0.001), crypt abscesses, vascular dilatation (p<0.01), and mucosal atrophy (p<0.05) between the OOEO-0.1mg/kg-IP and control colitis groups. No significant difference was determined in terms of ulceration, inflammatory cyst, mucosal atrophy, edema, and vascular dilatation between the dexamethazone and control colitis groups (p>0.05). Under the present conditions, we concluded that IR and IP OOEO treatment, applied at the dosage of 0.1 or 1mg/kg/day, have a significant protective effect on the colonic injury.

Composition, enantiomeric distribution, and antimicrobial activity of Tanacetum argenteum subsp. flabellifolium essential oil.

Tanacetum argenteum (Lam.) Willd. subsp. flabellifolium (Boiss. & Heldr.) Grierson of Asteraceae is an endemic species in Turkey. Hydrodistillation of aerial parts using a Clevenger apparatus yielded an essential oil, which was subsequently analyzed by gas chromatography-mass spectroscopy (GC-MS). alpha-Pinene (29%), (E)-sesquilavandulol (16%), and camphor (14%) were found as main constituents. Enantiomeric distribution of the monoterpenes alpha-pinene and camphor was determined on a fused silica Lipodex E capillary column using a multidimensional gas chromatography-mass spectroscopy (MDGC-MS) system, (-)-alpha-pinene (86%), (+)-alpha-pinene (14%), and (-)-camphor (100%) enantiomeric distributions were found in the oil. Furthermore, antimicrobial activity of the oil was carried out using a micro-dilution assay against human pathogenic bacteria and the yeast Candida albicans resulting in moderate inhibitory concentrations (MIC=125 microg/mL).

Gas chromatographic-mass spectrometric analysis of essential oils from Pimpinella species gathered from Central and Northern Turkey.

Essential oils from 15 Pimpinella species were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) techniques. One species, Pimpinella anisum, in which only fruits were evaluated, was also included in the study. A total of 140 different compounds were identified and significant qualitative and quantitative differences were observed among the samples. Pimpinella essential oils were characterized as having mono-, sesqui- and trinorsesquiterpenoids, propenylphenols, and pseudoisoeugenols. Trinorsesquiterpenoids and phenylpropanoids appear to be chemical markers of Pimpinella species analyzed thus far. Essential oils obtained from Pimpinella roots share the same principal compound, epoxypseudoisoeugenyl-2-methylbutyrate at concentrations from 20 to 82.6%.

The essential oil constituents and antimicrobial activity of Anthemis aciphylla BOISS. var. discoidea BOISS.

The essential oil of aerial parts, leaves and flowers of the endemic Anthemis aciphylla BOISS. var. discoidea BOISS. (Asteraceae) were obtained by hydrodistillation. The oils were analyzed both by GC and GC-MS on a polar column. The monoterpenes alpha-pinene (9-49%) and terpinen-4-ol (22-32%) were characterized as the main constituents. An unknown component isolated from the essential oil was characterized by means of MS, HR-MS, FT-IR, 1D- and 2D-NMR techniques as isofaurinone (1). Furthermore, the biological activity of the essential oils was evaluated in various human pathogenic microorganisms using the broth microdilution method. Weak to moderate inhibitions (0.06-1.0 mg/ml) was observed.

Gas chromatographic-mass spectrometric analysis of essential oils from Pimpinella aurea, Pimpinella corymbosa, Pimpinella peregrina and Pimpinella puberula gathered from Eastern and Southern Turkey.

Essential oils from fruits, stems and leaves and roots of Pimpinella aurea DC., P. corymbosa Boiss., P. peregrina L. were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) techniques. Fruits and aerial parts of P. puberula (DC.) Boiss were also evaluated. A total of 140 different compounds were identified, and significant qualitative and quantitative differences were observed among the samples. In fact, the main constituents of each species were different and only the oils extracted from roots shared the same principal compound, epoxy pseudoisoeugenyl-2-methyl butyrate (26.8-42.8%). The other fractions were dominated by different sesquiterpene compounds although in three of them, P. aurea stem and leaves, P. puberula fruits and P. puberula stems and leaves, monoterpene constituents also appear as main ones.

Estrogenic activity of isolated compounds and essential oils of Pimpinella species from Turkey, evaluated using a recombinant yeast screen.

Several plants and plant-derived pure compounds, designated as phytoestrogens, have been reported to cause estrogenic effects. They have been used for alleviation of menopausal symptoms, prevention of osteoporosis, heart disease and cancer. There is an increased interest in studying phytoestrogens such as isoflavones and lignans for their use as replacements for synthetic estrogens. In this study, the estrogenic activity of essential oils of eleven Pimpinella species and the compounds isolated from these species were evaluated using the yeast estrogen screen (YES) assay. The essential oils containing (E)-anethole as major compound showed estrogenic activity in the YES assay, except for the aerial parts without fruits of P. anisetum and P. flabellifolia. The percent maximal response produced by most anethole-containing oils was 30-50%. Fruits of P. isaurica and P. peucedanifolia were estrogenic in spite of the absence or trace amount of anethole, respectively. This study indicates that the estrogenic activity of Pimpinella oils is not solely due to the presence of anethole. Components other than anethole may be responsible for contributing towards the estrogenic activity. The essential oils from different species varied in their estrogenic potencies (relative potency from 8.3 x 10(-8) to 1.2 x 10(-6) compared to 17 beta-estradiol) and among the different plant parts, the fruit oils of most species were estrogenic followed by the aerial parts without fruits and the root oils and their EC50 values varied from 45 micrograms/mL to 650 micrograms/mL.

Investigation of the Origanum onites L. essential oil using the chorioallantoic membrane (CAM) assay.

The in vivo test on the chorioallantoic membrane of the fertilized hen's egg (CAM assay) is a current method to determine antiangiogenic, antiinflammatory activity and toxic effects of individual compounds or complex plant extracts. The method is used for testing natural compounds in small amounts for revealing various modes of action and the complex mechanisms related to angiogenesis and inflammation. Furthermore, possible side effects such as membrane irritation, toxic, and anticoagulant properties of the investigated material in question can be detected. For the evaluation, the essential oil obtained by hydrodistillation of the aerial parts of Origanum onites L., a common spice and medicinal plant, was tested for its effect in the chorioallantoic membrane (CAM) assay. The essential oil composition was revealed by means of gas chromatography-mass spectrometry (GC-MS). Eighty three components were identified, representing 99.1% of the total oil. Carvacrol, thymol, p-cymene, and gamma-terpinene were found as major components and were also individually tested in the CAM assay. Along with the monoterpenes carvacrol and thymol, their methyl ether derivatives were also examined for comparison of their physiological action. Neither the essential oil nor its components showed any pronounced antiinflammatory or antiangiogenic property in the CAM assay, at 10-250 microg/pellet. However, the irritant effect of the essential oil was linked to thymol in a dose-response fashion, up to 10 microg/pellet, where it was still showing irritation.

The bioactive essential oil of Heracleum sphondylium L. subsp. ternatum (Velen.) Brummitt.

The essential oil of Heracleum sphondylium L subsp. ternatun (Velen.) Brummit (Umbelliferae) was isolated from crushed seeds by means of hydrodistillation and analyzed by GC and GC/MS. Major components were identified as 1-octanol (50.3%), octyl butyrate (24.6%), and octyl acetate (7.3%). Furthermore, antimicrobial activity of the oil was evaluated using microdilution broth and agar diffusion methods. The bioactive constituent of the essential oil was determined as 1-octanol by using a bioautography assay.

Composition and antimicrobial activity of the essential oil of Achillea multifida.

The composition of the water-distilled essential oil of Achillea multifida (DC.) Boiss. (Compositae) was analysed by GC and GC/MS. Fifty-eight compounds were identified representing 93.9 % of the total oil. alpha-Thujone (60.9 %), beta-thujone (9.1 %), sabinene (4.1 %) and camphor (3.7 %) were characterised as the main constituents. The essential oil was tested for its antimicrobial activity using a micro-dilution assay resulting in the inhibition (MIC: 62.5-250 microg/ml) of human pathogenic bacteria and yeast.

Ferulagone: a new monoterpene ester from Ferulago thirkeana essential oil.

Essential oil obtained by micro-distillation and hydrodistillation of the endemic Ferulago thirkeana (Boiss.) Boiss. (Apiaceae) was analysed by GC/MS. An unknown component isolated from the essential oil was characterized by chromato-spectral techniques (1D-, 2D-NMR, HRMS, IR and UV) as (1 S)-2,6,6-trimethyl-4-oxobicyclo[3.1.1]hept-2-enyl(2 E)-2-methylbut-2-enoate (= ferulagone or 1-angeloyloxyverbenone) (1). Biological activities of the essential oil and the purified major component (1) were tested against various human pathogenic microorganisms resulting in moderate inhibition (62.5 - 125 microg/ml).