Study of potent cytotoxic activity of Helleborus cyclophyllus Boiss against a human adenocarcinoma cell line.

Helleborus cyclophyllus Boiss is a rhizomatous plant species, with strong allelochemical properties, that has been used since ancient times for its therapeutic properties. In the present study we investigated the ability of an aqueous-soluble fraction of the methanol extract of H. cyclophyllus Boiss leaves, to induce apoptotic cell death on A549 human bronchial epithelial adenocarcinoma cells. A primary human lung fibroblasts' cell line was used as a model of normal-healthy cells for comparison. Cell morphology was examined after appropriate staining, cytotoxic activity of the extract was determined by the MTT assay, the type of cell death was analyzed by flow cytometry, confirmation of apoptosis was evaluated with the analysis of caspase-3, PARP1 by western blotting, while the chemical composition was assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). H. cyclophyllus Boiss extract was selectively active on A549 cells inducing significant morphological changes, even at low concentrations. Characteristic morphological alterations included the release of vesicular formations from A549 cell membranes (ectosomes), detachment of cells from their substrate, generation of a large vesicle into the cytoplasm (thanatosome) and the formation of apoptotic bodies. The selective apoptotic action on treated cells was also confirmed by biochemical criteria. Low concentrations, however, did not affect normal cells. The phytochemical analysis of the extract revealed the presence of cardiac glucosides, bufadienolides and phytoecdysteroids. To the best of our knowledge, the above-mentioned sequences of events leading selectively cancer cells to apoptosis, has not been reported before.

Dissipation and transport of quizalofop-p-ethyl herbicide in sunflower cultivation under field conditions.

In the present study, the field dissipation and transport of quizalofop-p-ethyl by water and sediment runoff were investigated in sunflower experimental cultivation under Mediterranean conditions. The cultivation was carried out in silty clay soil plots with two different slopes of 1 and 5%. The soil dissipation rate of quizalofop-p-ethyl was fast and can be described by both single first-order (SFO) and Gustafson and Holden (first-order multi compartment (FOMC)) kinetics. The half-life of quizalofop-p-ethyl ranged from 0.55 to 0.68 days and from 0.45 to 0.71 days when SFO and FOMC kinetics were applied, respectively. No herbicide residues were detected below the 10-cm soil layer. A single detection of quizalofop-p-ethyl was observed in runoff water (3 days after application (DAA)) at relatively low concentrations (from 1.70 to 2.04 µg L(-1)). In sediment, it was detected in the samplings of 3 and 25 DAA at concentrations that never exceeded 0.126 µg g(-1). The estimated total losses of quizalofop-p-ethyl as percentage of the initial applied active ingredient were low both in water and sediment (less than of 0.021 and 0.005%, respectively). Quizalofop-p-ethyl residues were detectable for 18 DAA in the stems and leaves of the plants and 6 DAA in the root system. No herbicide residues were detected in inflorescences and seeds of sunflower plants. Experimental data showed minimal risk for the contamination of soil and adjacent water bodies.

Rosmarinic acid failed to suppress hydrogen peroxide-mediated apoptosis but induced apoptosis of Jurkat cells which was suppressed by Bcl-2.

Rosmarinic acid (RosA), frequently found as a secondary metabolite in herbs and medicinal plants, has exhibited antioxidative and anti-inflammatory activities. RosA was shown to inhibit the proliferation and induce apoptosis of Jurkat T cells but the mechanism of action of RosA in apoptosis remains elusive. RosA inhibited the proliferation of Jurkat cells in a dose-dependent manner by suppressing the expression of cyclin D3 and p21(Cip1/Waf1) and up-regulating p27(Kip1). RosA induced apoptosis of Jurkat cells in a dose-dependent manner and failed to protect them from hydrogen peroxide (H2O2)-mediated apoptosis. Induction of apoptosis by RosA correlated with suppression of Bcl-2 but not of Bak or PUMA. Overexpression of Bcl-2 protected Jurkat cells from both H2O2- and RosA-induced apoptosis by altering the ratio of anti- to pro-apoptotic members of the Bcl-2 family. In conclusion, RosA inhibited Jurkat cell proliferation by altering the expression of cyclins and cyclin-dependent kinase inhibitors and induced apoptosis most likely acting through the mitochondrial pathway and possessed no anti-oxidant properties.