Bioactivity of some Apiaceae essential oils and their constituents against Sitophilus zeamais (Coleoptera: Curculionidae).

Sitophilus zeamais is a key pest of stored grains. Its control is made, usually, using synthetic insecticides, despite their negative impacts. Botanical insecticides with fumigant/repellent properties may offer an alternative solution. This work describes the effects of Anethum graveolens, Petroselinum crispum, Foeniculum vulgare and Cuminum cyminum essential oils (EOs) and (S)-carvone, cuminaldehyde, estragole and (+)-fenchone towards adults of S. zeamais. Acute toxicity was assessed by fumigation and topical application. Repellence was evaluated by an area preference bioassay and two-choice test, using maize grains. LC50 determined by fumigation ranged from 51.8 to 535.8 mg L-1 air, with (S)-carvone being the most active. LD50 values for topical applications varied from 23 to 128 µg per adult for (S)-carvone > cuminaldehyde > A. graveolens > C. cyminum > P. crispum. All EOs/standard compounds reduced significantly the percentage of insects attracted to maize grains (65-80%) in the two-choice repellence test, whereas in the area preference bioassay RD50 varied from 1.4 to 45.2 µg cm-2, with cuminaldehyde, (S)-carvone and estragole being strongly repellents. Petroselinum crispum EO and cuminaldehyde affected the nutritional parameters relative growth rate, efficiency conversion index of ingested food and antifeeding effect, displaying antinutritional effects toward S. zeamais. In addition, P. crispum and C. cyminum EOs, as well as cuminaldehyde, showed the highest acetylcholinesterase inhibitory activity in vitro (IC50 = 185, 235 and 214.5 µg mL-1, respectively). EOs/standard compounds exhibited acute toxicity, and some treatments showed antinutritional effects towards S. zeamais. Therefore, the tested plant products might be good candidates to be considered to prevent damages caused by this pest.

Antigenotoxic effects of quercetin, rutin and ursolic acid on HepG2 cells: evaluation by the comet assay.

In the present study, the chemoprotective effects of quercetin, rutin and ursolic acid on tert-butyl hydroperoxide (t-BHP)-induced DNA damage in a human hepatoma cell line (HepG2) were investigated by the comet assay. To determine whether protection was due to direct chemical interactions alone or to cellular-mediated responses three different types of treatments were used: simultaneous incubation of cells with individual test compounds and the toxicant; pre-treatment with test compound before addition of the toxicant followed or not by a recovery period. The expression of Hsp70 was quantified by Western blotting to test the involvement of heat shock proteins in the cellular responses to the test compounds. In addition, effects on proliferation were evaluated by the MTT assay. The results show that quercetin and ursolic acid prevented DNA damage and had antiproliferative properties in HepG2 cells suggesting an anticarcinogenic potential for these compounds. The protective effects of quercetin against t-BHP-induced DNA damage seem to be due to both direct effects on t-BHP toxicity and to cellularly mediated indirect effects which reflect the potentiation of the cellular antioxidant defenses. Ursolic acid seems to exert effects only through cellularly mediated mechanisms since it was not protective in simultaneous incubation. Quercetin and ursolic acid also showed to increase the rate of DNA repair. Rutin did not have effects at any level. These results, obtained with liver cells, emphasize and confirm the chemopreventive potential of quercetin and ursolic acid, which may help explain the lower cancer incidence in human population with high dietary intakes of fruits and vegetables. These results also demonstrate that Hsp70 is not involved in the observed effects in HepG2.

Variation of the essential oil content and composition in leaves from cultivated plants of Hypericum androsaemum L.

The amount and composition of the essential oil from leaves of Hypericum androsaemum L. cultivated in Arouca (Portugal) were determined in six samples harvested during 1 year at intervals of 2 months. The seasonally dependent essential oil content ranged from 0.7 mg/g biomass dry weight in September to 3.4 mg/g in February. The oil contained more than 80 compounds, 70 of which (constituting 88-93% of the total oil) were identified by GC and GC-MS. An approximation of the absolute quantification of each compound and compound class was performed using a GC method with an internal standard. The relative and the absolute content of each compound and compound class changed during the year. At the end of the winter and in the spring, the essential oil was dominated by sesquiterpene hydrocarbons and accumulated a high number of intermediate to long chain n-alkanes and 1-alkenes. In September, the essential oil contained the lowest levels of sesquiterpene hydrocarbons (43%) and the highest levels of 1-octene and 2-hexenal (38%). In February, the essential oil had the highest level of sesquiterpene hydrocarbons (73%) and the highest diversity of intermediate to long chain n-alkanes and 1-alkenes.

Evaluation of toxic/protective effects of the essential oil of Salvia officinalis on freshly isolated rat hepatocytes.

For this study the essential oil (EO) of sage (Salvia officinalis L.) was isolated from air-dried vegetative aerial parts of the plants by hydrodistillation and analysed by GC and GC-MS. A total yield of 12.07 mg of EO per g of plant dry mass was obtained and more than 50 compounds identified. The major compounds were cis-thujone (17.4%), alpha-humulene (13.3%), 1,8-cineole (12.7%), E-caryophyllene (8.5%) and borneol (8.3%). The EO fraction of sage tea was also isolated by partition with pentane and the respective components identified. The toxic and antioxidant protective effects of S. officinalis EO were evaluated on freshly isolated rat hepatocytes. Cell viability (LDH leakage), lipid peroxidation and glutathione status were measured in experiments undertaken with cells (suspensions of 1 x 10(6) cells per millilitre) exposed to EO alone (toxicity of the EO;t-BHP as positive control); and with cells exposed to EO and an oxidative compound (t-BHP) together (in EO protection evaluation; quercetin as positive control) for 30 min. The results show that the EO is not toxic when present at concentrations below 200 nl/ml; it was only at 2000 nl EO/ml that a significant LDH leakage and GSH decrease were observed indicating cell damage. In the range of concentrations tested, the EO did not show protective effects against t-BHP-induced toxicity.

Organ- and season-dependent variation in the essential oil composition of Salvia officinalis L. cultivated at two different sites.

More than 50 compounds were identified in essential oils from stems and leaves of Salvia officinalis L. plants harvested in July, in Arouca, in northern Portugal. About 40 of those compounds were also present in flower essential oils, collected from the same plants. alpha-Thujone was the major compound, representing about 55, 30, and 18% of the essential oils from stems, leaves, and flowers, respectively. Significant percentage variations in the main compound classes of the essential oils from shoots sampled over the year were recorded at two different sites in northern Portugal. From December to April, oxygenated monoterpenes (MO) decreased from approximately 67-72% to values of 42-43% of the essential oils. During the same time interval, the percentage of monoterpene hydrocarbons (MH) rose from 8-11% to 17-22%. At both sites, sesquiterpene hydrocarbons (SH) rose from approximately 7% in February to 19-22% in April, decreasing thereafter to approximately 9% in July. Oxygenated sesquiterpenes (SO) increased from a minimum of approximately 5% in July to a maximum of 8-11% in February, decreasing thereafter. The compounds that mostly accounted for the essential oil composition variation were alpha-pinene, beta-pinene, and camphene, as MH; alpha-thujone and camphor, as MO; alpha-humulene and beta-caryophyllene, as SH; and viridiflorol, as SO.

Ontogenic variations in n-alkanes during somatic embryogenesis of flax (Linum usitatissimum L.).

Hypocotyl segments (HS) of flax seedlings germinated in vitro, were used to induce indirect somatic embryogenesis on solid medium. The composition and distribution of n-alkanes in flax tissues collected at different developmental stages were studied by capillary gas chromatography (GC) and capillary gas chromatography-mass spectrometry (GC-MS). During induction and development of callus from hypocotyl tissues a decrease in the percentage of total lipids was observed. In all types of tissue sampled - HS used as primary explants, HS with differentiating calli at the cut ends (HSC), embryogenic (EC) and non-embryogenic calli (NEC) and somatic embryos (SE) - a skewed-normal distribution of n-alkanes with a low mass range (C(13)-C(21)) were found. The highest content of n-alkanes occurred in the primary hypocotyl explants and in the early stages of callus development. Longer carbon chain n-alkanes were observed only in the mature or differentiated tissues of hypocotyls and SE. Although the n-alkane contents decreased with time, in SE and calli, a significantly lower n-alkane content was observed in EC when compared to NEC independent of the time in culture. These results suggest the utilisation of n-alkanes for heterotrophic cellular growth as well as its mobilisation from EC to developing SE.