Effects of pine needle extracts on plasma cholesterol, fibrinolysis and gastrointestinal motility

Pine needle (Pinus densiflora Sieb. et Zucc.) extract has been used to improve cardiovascular disorders, detoxification of nicotine, the infirmities of age and curing diseases of unidentified symptoms in folk medicine. To determine the facts behind the traditional belief, we tried to investigate the effects of fresh and self-fermented pine needle extracts of different aging. Fibrinolytic activities of the extract indicated that activity depends on time and also with aging of the product. It was also found that the extract can lower the blood plasma cholesterol and triglyceride in cholesterol fed rat. Also, Self-Fermented Pine Needle Extracts 7 years old (SFPE 7) (200 μg/mL) reduce the frequency and amplitude of pacemaker currents in Interstitial Cells of Cajal (ICC) of murine small intestine by modulating ATP-sensitive potassium channels. Therefore, the investigation indicated that self-fermentation improves efficacy of the pine needle extracts reducing risk of cardio-vascular related disorders and would be an important source in nutraceutics.     

Rosmarinus officinalis L. essential oils from Spain: composition,antioxidant capacity,lipoxygenase and acetylcholinesterase inhibitory capacities,and antimicrobial activities

Abstract

Six Rosmarinus officinalis L. essential oils (RoEOs) from Murcia (Spain) were studied using gas chromatography. Analysis of their relative and absolute composition showed 1,8-cineole, camphor, α-pinene and camphene to be the main compounds. Moreover, enantioselective gas chromatographies showed different enantiomeric ratios of camphor, limonene and borneol. The antioxidant and chelating capacities of RoEOs and their individual components were measured in an attempt to ascertain the cause of RoEO activity as a whole. All the essential oils were able to inhibit lipoxygenase activity due to bornyl acetate, camphor, terpinen-4-ol and 1,8-cineole and acetylcholinesterase activity due to 3-carene and 1,8-cineole, mainly. When the antimicrobial activities were tested against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Candida albicans, the least sensitive microorganism was P. aeruginosa and the most sensitive C. albicans. This study furthers our knowledge of six RoEOs from two chemotypes, all of which show potential for industrial applications.     

Screening of antibacterial activities of twenty-one oxygenated monoterpenes

Plant essential oils are widely used as fragrances and flavours in the cosmetic, perfume, drug and food industries. Oxygenated monoterpenes are widespread components of the essential oils, usually occurring in high amount. In this paper, the antibacterial activities of twenty-one oxygenated monoterpenes (borneol, borneol acetate, camphor, carvone, 1,8-cineole, citronellal, beta-citronellol, dihydrocarvone, fenchol, fenchone, geraniol acetate, isomenthol, limonene oxide, linalool, linalool acetate, nerol, nerol acetate, terpinen-4-ol, alpha-terpineol, menthol and menthone) and penicillin (standard antibiotic) were determined using a disc diffusion method (in vitro) against 63 bacterial strains, belonging to 37 different genera and 54 species (plant, food and clinic origins). The results showed that the oxygenated monoterpenes exhibited a variable degree of antibacterial activities. These compounds also inhibited the growth of bacterial strains by producing a weak zone of inhibition from 7 to 11 mm in diameter, depending on the susceptibility of the tested bacteria. Among the tested compounds, nerol, linalool alpha-terpineol, fenchol and terpinen-4-ol showed antibacterial activity at a broad spectrum. However, their antibacterial activities were lower than those of penicillin. In contrast to these compounds, camphor and 1,8-cineole exhibited no inhibition effects on the growth of all tested bacteria.     

RNAi targeting the gene for 1,8-cineole synthase induces recomposition of leaf essential oil in lavandin (Lavandula × intermedia Emeric)

Recomposition of volatile compounds is an effective way to alter the fragrance of an essential oil. Using RNA interference (RNAi), we attempted to reduce the production of 1,8-cineole, a major monoterpene in essential oils, to alter the composition of the essential oil in lavandin. We obtained 12 transgenic regenerants via inoculation with Agrobacterium tumefaciens, including an RNAi inducing vector targeting five regions of the gene for 1,8-cineole synthase (CINS). Of these, two regenerants, targeting a coding region about 250 nt upstream with 5′-UTR and a coding region about 300 nt downstream of CINS mRNA (CINS I and V), respectively, showed a significant decrease of 1,8-cineole production in leaf essential oil, although the overall composition was barely altered because the production of other compounds decreased concurrently. By contrast, in two other regenerants, targeting a coding region about 1000 nt in the middle and a coding region about 300 nt downstream of CINS mRNA (CINS IV and V), respectively, 1,8-cineole production could be barely observed, without any decrease in production of other compounds. Expression of CINS in these transgenic regenerants was extremely suppressed to 0.02 and 0.08 of that of a nontransgenic regenerant control. The composition of leaf essential oil in the transgenic regenerants was changed by the RNAi. Specifically, the major compounds changed from 1,8-cineole, camphor, and borneol, to linalool, camphor, and borneol. Consequently, the fragrance of essential oils of these plants was perceived as more citrusy than the fragrance of the nontransgenic regenerant. These results suggest that the knockdown strategy was a useful tool for altering the fragrance in lavandin.

     

The inflorescence and leaf essential oils of Tanacetum vulgare L. var. vulgare growing wild in Lithuania

Forty samples of inflorescences and leaves of wild Tanacetum vulgare L. var. vulgare were collected in 20 habitats from Lithuania. The essential oils were analyzed by GC and GC/MS. The 57 identified compounds in the oils made up 80.7–99.6%. According to the cluster analysis the volatile oils were divided into four groups with 1,8-cineole (23.6–46.3%, 11 oils), trans-thujone (35.7–78.4%, 6 samples), camphor (19.8–61.8%, 17 oils) and myrtenol (13.1–24.9%, 6 samples) as main constituents. The inflorescences and leaves of tansy plants formed the oils with the same dominating constituent in 15 of the 20 habitats investigated. The leaves in five localities produced oils of the 1,8-cineole chemotype, while the inflorescences biosynthesized oils of the camphor type in three habitats and of the myrtenol type in two habitats. Amounts of the 1,8-cineole in all leaf oils were greater than that in inflorescence oils of the plants from the same locality. An opposite correlation was determined for camphor, myrtenol, cis- and trans-thujone. The myrtenol chemotype was not noticed earlier in the essential oils of T. vulgare.     

Identification of pheromone candidates for the eucalyptus weevil,Gonipterus platensis (Coleoptera,Curculionidae)

The eucalyptus weevil, Gonipterus platensis (Coleoptera, Curculionidae), is a major pest of eucalyptus plantations worldwide. To date, no pheromones have been identified for this species, despite their valuable potential as tools in monitoring or control strategies. Here we report the detection and identification of pheromones candidates of G. platensis. The weevil's volatile compounds were collected by solid phase micro extraction (SPME) and monolithic material sorption extraction (MMSE). Using Gas Chromatography coupled to Mass Spectrometry (GC/MS) analysis, eleven insect specific compounds were detected and identified: verbenene, cis-verbenol, trans-verbenol, verbenone, 2-oxo-1,8-cineole, 9-hydroxy-1,8-cineole, 2-α-hydroxy-1,8-cineole, 3-oxo-1,8-cineole, 2-β-hydroxy-1,8-cineole, 3-α-hydroxy-1,8-cineole and 7-hydroxy-1,8-cineole. Three of these compounds, verbenene, cis-verbenol and trans-verbenol, were shown to be male-specific. Antennal sensitivity towards ten compounds emitted by G. platensis was detected using Gas Chromatography–Mass Spectrometry/Electroantennographic Detection (GC-MS/EAD). Extracts from virgin males proved to be attractive to virgin females in olfactometer bioassays. Further behavioural bioassays showed that both virgin females and virgin males were attracted to the male-specific compound cis-verbenol and that virgin females were attracted to trans-verbenol. Verbenone was attractive to mated females. Regarding 2-α-hydroxy-1.8-cineole and 2-oxo-1,8-cineole, which are produced by both sexes, the alcohol was attractive to virgin males and both the alcohol and the ketone were repellant to mated females. This is, to our knowledge, the first identification of pheromones candidates in Gonipterus spp. and also the first evidence of cineole metabolites acting as semiochemicals.     

Enhanced antioxidant and antitumor activities of Antrodia cinnamomea cultured with cereal substrates in solid state fermentation

Growing exclusively on stout camphor trees in Taiwan, Antrodia cinnamomea is known for its extraordinary antioxidant and antitumor activities. As an alternative to the limited supply of natural source, cultured A. cinnamomea from solid state or submerged liquid fermentation still offers many of its medicinal effects. To further enhance the production of functional compounds and corresponding activities, oat, wheat, buckwheat and pearl barley were used as substrates for solid state fermentation of A. cinnamomea in this study. Among these cereal-based culturing, the methanol extract of A. cinnamomea mycelia grown on oats showed stronger overall antioxidant properties. EC₅₀ for the antioxidant activity (conjugated diene method), the DPPH radicals scavenging ability and reducing power were estimated to be around 0.57 mg/mL, 1.07 mg/mL and 0.31 mg/mL, respectively. Incubating cultured cells with 150 ppm of the oat-cultured mycelial extract for 24 h greatly reduced the viability of MCF-7 breast cancer cells and HepG2 hepatocellular carcinoma cells to 29% and 76%, while 3T3 normal fibroblasts were virtually unaffected. In general, cereal-based solid state fermentation of A. cinnamomea produced more of the secondary metabolites and their methanolic extracts showed stronger antioxidant and anti-tumor activities than extracts obtained from liquid fermentation at the same concentration.     

Volatiles from spruce trap-trees detected by Ips typographus bark beetles: chemical and electrophysiological analyses

In the search for compounds that contribute to the host or habitat discrimination, antennae of Ips typographus were screened for sensitivity to volatiles released by spruce trap-trees using gas chromatography linked to electroantennography. The antennally active compounds were determined using comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometric detection. Data show that I. typographus antennae respond to compounds emitted by the host. In total, 18 of antennally active compounds were detected: α-pinene, camphene, sabinene, β-pinene, myrcene, Δ-3-carene, p-cymene, limonene, β-phellandrene, 1,8-cineole, γ-terpinene, terpinolene, nonanal, camphor, trans-pinocamphone, cis-pinocamphone, terpinen-4-ol, and verbenone. Unequivocal identification of all active minor compounds is provided and confirmed using synthetic standards. Compounds in minor quantities like 1,8-cineole, β-phellandrene, camphor, cis-pinocamphone, and trans-pinocamphone were more active than major spruce monoterpenes. We hypothesize that the minor spruce compounds may play so far unrecognized role in conveying information about host suitability for I. typographus.     

Chemical composition of essential oils of four Eucalyptus species and their phytotoxicity on silverleaf nightshade (Solanum elaeagnifolium Cav.) in Australia

Phytotoxicity and chemical composition of essential oils from four selected Eucalyptus species in Australia were investigated. Essential oils had stronger inhibitory effects on germination and seedling growth of silverleaf nightshade (Solanum elaeagnifolium Cav.) when compared with a commercial eucalyptus oil and with 1,8-cineole. E. salubris oil had the highest inhibition index for silverleaf nightshade germination, root growth and shoot growth, while E. spathulata had the lowest inhibitory effect except root growth. Gas chromatography-mass spectrometry analysis revealed 56 compounds present in E. salubris oil, with 1,8-cineole (57.6?%), ??-pinene (10.9?%) and p-cymene (8.3?%) predominant. E. dundasii oil contained 55 identified compounds with 1,8-cineole (65.5?%) and ??-pinene (19.9?%) being the richest fractions. There were 56 compounds identified from E. brockwayii oil with ??-pinene (31.1?%), isopentyl isovalerate (20.2?%) and 1,8-cineole (16.9?%) as the most abundant components. E. spathulata oil contained 60 compounds, predominantly 1,8-cineole (52.9?%) and ??-pinene (31.0?%). Further study is required to determine the phytoxicity of the individual identified compounds on silverleaf nightshade and whether the observed phytotoxicity is attributable to a single compound or to the synergistic effects of several compounds.     

Variation of Salvia fruticosa Essential Oils on the Island of Crete (Greece)

The essential oil content and composition of Salvia fruticosa (Greek sage) plants growing wild in 20 localities scattered on the island of Crete are studied. The results of our analyses have shown a noticeable variation in the essential oil content (ranging from 1.1 up to 5.1 %) and the amount of the four main oil components: 1,8-cineole (22.7 ? 64.2% of total oil), α-thujone (1.0 ? 19.2%) β-thujone (0.9 ? 25.6%) and camphor (0.8 ? 30.3%). Discriminant analysis revealed that the variation pattern of the essential oil content and the amount of the four main oil components is geographically related, following a W → E direction. Plants grown in Western Crete show a lower essential oil content and their oils are characterised by the predominance of 1,8-cineole. On the other hand, those collected from Eastern Crete exhibit higher values in essential oil content and their oils, besides 1,8-cineole, are rich in α- and β-thujone or camphor. Our findings are further discussed in relation to literature data.     

Anti-oxidant activity and effect of Pinus morrisonicola Hay. on the survival of leukemia cell line U937

The free radical scavenging and anti-cancer activites of Pinus morrisonicola Hay. were studied using different parts of the pine, namely, needle, bark and cone. Results showed that pine needle water extract has the highest scavenging superoxide anion activity and the lowest IC50 value in inhibiting superoxide anion formation; however, the bark water extract showed the best anti-lipid peroxidation activity. Additionally, needle water extract displayed the highest inhibition of leukemia cell line U937 growth. The results indicated that P. morrisonicola Hay. possesses potential chemopreventative and therapeutic properties.     

1,8-cineole and castor oil in sodium lauryl ether sulphate disrupt reproduction and ovarian tissue of Rhipicephalus (Boophilus) microplus

Essential and fixed oils have been researched as alternatives to chemical acaricides. The activity of volatile compounds from essential oils (1,8-cineole, citral and eugenol) at 1.0% (w/v) and fixed oil (castor oil) at 0.3% (w/v) dissolved in 2.0% (v/v) dimethyl sulfoxide (DMSO) + 0.2% (w/v) Tween 80^® was assessed against Rhipicephalus microplus using immersion tests. 1,8-cineole (29.0%) and castor oil (30.2%) had the highest reproductive inhibition rate. A second experiment was performed to verify the effect of the 1,8-cineole (10.0% w/v) and, or castor oil (0.3% w/v) on tick reproduction using different solubilizing agents. The highest reproductive inhibition was observed for the combination of 1,8-cineole/castor oil (94.1%) and 1,8-cineole in 2.0% (w/v) sodium lauryl ether sulphate (SLES) (92.8%). A third experiment showed morphological changes in R. microplus oocytes at different stages of development, as well as in pedicel cells. The most intense effects were observed when ticks were immersed in the formulation containing 1,8-cineole (10.0% w/v) and castor oil (0.3% w/v) dissolved in 2% (w/v) SLES. These findings highlight the potential of this formulation as an alternative for managing cattle ticks as their cytotoxic effects can reduce R. microplus reproductive success.     

Effect of monoterpenes on lipid oxidation in maize

The monoterpenes 1,8-cineole, thymol, geraniol, menthol and camphor strongly inhibited the root growth of Zea mays L. seedlings. They induced an oxidative stress as measured by the production of malondialdehyde, conjugated dienes and peroxides. This oxidative stress depended on the length of the exposure and on the monoterpene applied. The total fatty acid content was measured and fatty acid composition was analyzed. Unsaturated fatty acids increased in the treated samples. The alcoholic and non-alcoholic monoterpenes appeared to have different modes of action.Abbreviations MDA Malondialdehyde - TFA Total fatty acid content - FA Fatty acid - IC₈₀ Concentration causing 80% inhibition     

Chemical composition, antioxidant and antimicrobial properties of the essential oils of three Salvia species from Turkish flora

Essential oils of three different Salvia species [Salvia aucheri var. aucheri (endemic), Salvia aramiensis and Salvia pilifera (endemic)] were screened for their possible antioxidant and antimicrobial properties as well as their chemical compositions. According to the gas chromatography (GC)/EIMS (gas chromatography/electron impact mass spectrum) analysis results; 41 (97.2%), 51 (98.5%) and 83 compounds (98.2%) were identified, respectively. While 1,8-cineole (30.5%), camphor (21.3%) and borneol (8.50%) are the major compounds for S. aucheri var. aucheri oil, beta-pinene (10.3%), was the main constituent for S. aramienesis together with 1,8-cineole (46.0%) and camphor (8.7%). In the case of S. pilifera oil, alpha-thujene (36.1%) and alpha-pinene (13.8%) determined as the major compounds. Antioxidant activity was employed by two complementary test systems namely 2,2'-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and beta-carotene/linoleic acid systems. Antioxidant activity of S. aramiensis was found to be higher than those of the others for the both systems (12.26+/-1.09 and 92.46%+/-1.64 microg mg(-1), respectively). Additionally, antioxidant activities of BHT, curcumin, ascorbic acid and alpha-tocopherol were determined in parallel experiments. In the case of antimicrobial activity, similar activity pattern was obtained (both in disc diffusion and MIC tests). Antimicrobial activity of S. aramiensis was followed by S. aucheri var. aucheri and S. pilifera, respectively. In these experiments, the most sensitive microorganism Acinetobacter lwoffii was followed by Candida albicans.     

Composition of essential oils from turkish Salvia species

The chemical composition of the essential oils from five Salvia species from Turkey was determined by GC. The species were S. candidissima, S. cryptantha, S. fruticosa, S. officinalis and S. tomentosa. 24, 22, 20, 19, and 22 components were identified, respectively, the major ones being β-pinene (candidissima), borneol (cryptantha), 1,8-cineole (fruticosa), camphor (officinalis) and β-pinene (tomentosa). α-Pinene in candidissima, camphor in cryptantha, α-thujone in officinalis and 1,8-cineole in tomentosa were the other important components.     

Investigation of monoterpenoids rich essential oils of two Ocimum basilicum L. varieties at different agro-climatic conditions in India

India is an emerging basil essential oil producer in South-east Asia. Two high essential oil yielding hybrids, namely one inter specific hybrid between of O. basilicum and O. kilimandscharicum Gürke (HYBL-1) and another intraspecific hybrid of O. basilicum × O. basilicum (OBL-1) of basil were analyzed using GC, enantiomeric GC, NMR, enantio-GC–MS and GC–MS methods. Inter specific hybrid HYBL-1 contained high essential oil-rich in linalool (68.5%), camphor (8%), and 1,8-cineole (4.6%) as characteristic constituents among monoterpenoids, whereas β-caryophyllene (1.9%), germacrene D (1.0%), and epi-α-cadinol (1.9%) were the sesquiterpenoids at the Lucknow (North Indian conditions) and linalool (71.8%), camphor (9.4%) and 1,8-cineole (4.3%) at Hyderabad (South Indian conditions) locations. Intraspecific hybrid (OBL-1) possessed linalool (66.1%), 1,8-cineole (5.4%) and geraniol (8.6%) with sesquiterpenoids in low proportions. Inter specific hybrid HYBL-1 showed superiority over OBL-1 in the multi-location trials conducted at Lucknow and Hyderabad. Average mean performance of inter specific hybrid over locations was: herb yield 44.80 t/ha, oil content 0.63%, oil yield 188.50 kg/ha, linalool content 67.65%, camphor content 8.90% v/s OBL-1 herb yield 21.32 t/ha, oil content 0.53%, oil yield 97.50 kg/ha, linalool content 65.55%, camphor content 0.00%, respectively. The essential oil of these two hybrids subjected to enantiomer differentiation revealed a high enantiomeric excess for (3R) -(?)-linalool, whereas (1R)- (+)-camphor was recorded exclusively in inter specific hybrid. The extensive NMR experiments were performed to confirm constituents in these hybrids and found that NMR spectroscopy could also be an ideal tool for the differentiation of essential oils from commercial samples declared as natural.     

Anti-fungal activity of essential oils of Ceylon Eucalyptus species for the control of Fusarium solani and Sclerotium rolfsii

The essential oils were extracted from the leaves of Eucalyptus microcorys, Eucalyptus grandis and Eucalyptus robusta which were grown in Sri Lanka and their major chemical compounds were determined. 1,8-Cineole and α-pinene were identified as major aroma compounds in these oils. In this study, the anti-fungal activity of essential oils of E. microcorys, E. grandis and E. robusta, ethanol extract of E. microcorys and 1,8-cineole were evaluated against Sclerotium rolfsii, a fungi responsible for leaf spot disease of indoor plants and Fusarium solani, a fungi responsible for dry rot diseases of potato by poisoned food technique, and minimum inhibitory concentrations (MICs) were determined. The essential oils from three Eucalyptus species showed significant inhibitory effect against S. rolfsii and F. solani than the ethanol extract of E. microcorys. Of treatments, the essential oil of E. grandis showed the best anti-fungal activity with the MIC values of less than 0.1% for S. rolfsii and 0.5% for F. solani. The MICs of the oils of E. microcorys and E. robusta were between 0.3–0.5% against S. rolfsii and 0.5–0.75% for F. solani. The 1,8-cineole did not exhibit inhibition activity as much of Eucalyptus essential oils and hence, it can be assumed that minor chemical components of the oils contribute to the growth inhibition of the tested fungi. This is the first report of anti-fungal activity of Sri Lankan oils of E. microcorys, E. grandis and E. robusta and ethanol extract of E. microcorys against S. rolfsii and F. solani. These findings would be useful for the designing of natural fungicide for agriculture- and food-based industries.     

Environment-related variations of the composition of the essential oils of rosemary (Rosmarinus officinalis L.) in the Balkan Penninsula

Composition of the essential oils of Rosmarinus officinalis of ten populations from the Balkan Peninsula were determined by GC/FID and GC/MS. The main constituents were 1,8-cineole, camphor, α-pinene, and borneol. Multivariate statistical analysis (UPGMA cluster analysis and principal-component analysis (PCA)) revealed two major types of rosemary oil, i.e., 1,8-cineole and camphor-type, and two intermediate types, i.e., camphor/1,8-cineole/borneol type and 1,8-cineole/camphor type. The regression analyses (simple linear regression and stepwise multiple regression) have shown that, with respect to basic geographic, orographic, and 19 bioclimatic characteristics of each population, bioclimatic factor temperature of habitat represented the dominant abiogenetic factor, which, in chemical sense, led to differentiation of populations in the studied region. Also, the regression analysis have shown that some constituents of essential oils are independent of any single bioclimatic factors. However, some constituents display statistically significant correlations with some abiotic factors.