Topoisomerase poisoning by the flavonoid nevadensin triggers DNA damage and apoptosis in human colon carcinoma HT29 cells.

Nevadensin, an abundant polyphenol of basil, is reported to reduce alkenylbenzene DNA adduct formation. Furthermore, it has a wide spectrum of further pharmacological properties. The presented study focuses the impact of nevadensin on topoisomerases (TOPO) in vitro. Considering the DNA-intercalating properties of flavonoids, first, minor groove binding properties (IC50 = 31.63 µM), as well as DNA intercalation (IC50 = 296.91 µM) of nevadensin, was found. To determine potential in vitro effects on TOPO I and TOPO IIα, the relaxation and decatenation assay was performed in a concentration range of 1-500 µM nevadensin. A partial inhibition was detected for TOPO I at concentrations  ≥ 100 µM, whereas TOPO IIα activity is only inhibited at concentrations  ≥ 250 µM. To clarify the mode of action, the isolating in vivo complex of enzyme assay was carried out using human colon carcinoma HT29 cells. After 1 h of incubation, the amount of TOPO I linked to DNA was significantly increased by nevadensin (500 µM), why nevadensin was characterized as TOPO I poison. However, no effects on TOPO IIα were detected in the cellular test system. As a subsequent cellular response to TOPO I poisoning, a highly significant increase of DNA damage after 2 h and a decrease of cell viability after 48 h at the same concentration range were found. Furthermore, after 24 h of incubation a G2/M arrest was observed at concentrations ≥ 100 µM by flow cytometry. The analysis of cell death revealed that nevadensin induces the intrinsic apoptotic pathway via activation of caspase-9 and caspase-3. The results suggest that cell cycle disruption and apoptotic events play key roles in the cellular response to TOPO I poisoning caused by nevadensin in HT29 cells.

Oak Wood Volatiles Impact on Red Wine Fruity Aroma Perception in Various Matrices.

This research examined the impact of oak wood volatile compounds on the perception of red wine fruity aroma in several matrices. Several aromatic reconstitutions were prepared, consisting of 13 esters, representing the fruity pool of red wine, and 14 oak wood compounds at the various concentrations corresponding to the levels released by light, medium, and heavy toasting of barrels. These reconstitutions were prepared in dilute alcohol solution, dearomatized red wine, and commercial red wine. Sensory analysis revealed the impact of the addition or omission of some oak wood compounds. The "detection threshold" of the fruity pool was then evaluated. The presence of 2-furanmethanethiol individually and the oak wood compound mixture, at concentrations representing various toasting levels, had a significant masking effect on the fruity pool, whereas vanillin had a significant enhancing effect in model solution. Sensory profiles highlighted changes in the perception of fruity nuances in the presence of the oak wood compound mixture. The addition of compounds at concentrations representing different toasting levels led to a predictable increase in the perception of the oak wood descriptors (spicy, smoky, and toasty) in all of the matrices tested. The perception of fruity notes also varied depending upon the toasting level and the complexity of the matrix. In dilute alcohol solution and dearomatized red wine, light toasting preserved or intensified the fruity notes. Generally, in all matrices tested, fresh-fruit and red-berry-fruit notes decreased with the addition of wood at medium and heavy toasting levels.