Sonoporation of adherent cells under regulated ultrasound cavitation conditions.

A sonoporation device dedicated to the adherent cell monolayer has been implemented with a regulation process allowing the real-time monitoring and control of inertial cavitation activity. Use of the cavitation-regulated device revealed first that adherent cell sonoporation efficiency is related to inertial cavitation activity, without inducing additional cell mortality. Reproducibility is enhanced for the highest sonoporation rates (up to 17%); sonoporation efficiency can reach 26% when advantage is taken of the standing wave acoustic configuration by applying a frequency sweep with ultrasound frequency tuned to the modal acoustic modes of the cavity. This device allows sonoporation of adherent and suspended cells, and the use of regulation allows some environmental parameters such as the temperature of the medium to be overcome, resulting in the possibility of cell sonoporation even at ambient temperature.

Exposure of juvenile Danio rerio to aged TiO2 nanomaterial from sunscreen.

The toxicity of dietary exposure to artificially aged TiO2 nanomaterial (T-Lite) used in sunscreen cream was studied on Danio rerio. Embryolarval assays were conducted to assess the effects of TiO2 residues of nanomaterial (RNM) on fish early life stages. Juvenile fishes were exposed by the trophic route in two experiments. During the first experiment, juvenile fishes were exposed to TiO2 RNM for 14 days by adding RNM to commercial fish food. The second one consisted in producing a trophic food chain. Pseudokirchneriella subcapitata algae, previously contaminated with TiO2 RNM in growth medium, was used to feed Daphnia magna neonates over a 48-h period. Daphnia were used next to feed juvenile fishes for 7 days. Accumulation of Ti, life traits (survival and growth) and biochemical parameters such as energy reserves, digestive (trypsin, esterase, cellulose and amylase) and antioxidant (superoxide dismutase and catalase) enzyme activity were measured at the end of exposures. As expected in the receiving aquatic system, TiO2 RNM at low concentrations caused a low impact on juvenile zebrafish. A slight impact on the early life stage of zebrafish with premature hatching was observed, and this effect appeared mainly indirect, due to possible embryo hypoxia. When juvenile fish are exposed to contaminated food, digestive enzyme activity indicated a negative effect of TiO2 RNM. Digestive physiology was altered after 14 days of exposure and seemed to be an indirect target of TiO2 RNM when provided by food.

Effects of aged TiO2 nanomaterial from sunscreen on Daphnia magna exposed by dietary route.

The toxicity of dietary exposure to artificially aged TiO(2) nanomaterial (T-Lite™) used in sunscreen cream was studied on D. magna. Pseudokirchneriella subcapitata cultures were contaminated with TiO(2)-residues, obtained by artificial aging. Significant association of TiO(2)-residues on algae was detected by X-ray fluorescence spectromicroscopy. A D. magna dietary chronic exposure of these contaminated algae with TiO(2)-residues was performed. X-ray chemical imaging revealed that Ti was localized only in the digestive tract of the daphnia. Chronic exposure of daphnia to by-product of aged TiO(2) nanoparticles brought by food induced low mortality but decreased growth and reproduction which can be partly related to the modification of the digestive physiology of daphnia. This study demonstrated that the assessment of the ecotoxicological impact of nanomaterials in aquatic environment should take into account the aging of these materials which can further influence their bioavailability for aquatic organisms.

Dynamics of short-term acclimation to UV radiation in marine diatoms.

In order to investigate the dynamics of the acclimation of marine diatoms to ultraviolet radiation (UVR), Amphora coffeaeformis, Odontella aurita and Skeletonema costatum were exposed for 5 h per day to a combination of UVA and UVB (UVBR/UVAR ratio 4.5%) with a total UVR daily dose of 110 kJ m(-2), which is equivalent to that observed in the natural environment. This treatment was applied in the middle of the photoperiod and was repeated on five successive days. During the UVR treatment, chlorophyll fluorescence parameters were monitored, damage and repair constants were calculated from effective quantum yield values (phi(PSII)), and rapid light curves (electron transport rate versus irradiance curves using short light steps of different intensity) were plotted to determine the maximum relative electron transport rate (rETR(max)) and maximum light use efficiency (alpha). In all species the growth rate was lower than control from day 1-3, but increased thereafter, except for S. costatum. The cellular chlorophyll a content increased significantly with repeated daily exposure to UVR for A. coffeaeformis only. In all species, the fluorescence parameters (F(m), the maximum fluorescence level measured in the dark, phi(PSII), rETR(max) and alpha) decreased during UVR exposure, in contrast to F(0) (the minimum fluorescence level measured in the dark). The response to UVR stress was species-specific. S. costatum was very sensitive, and failed to survive for more than three days, whereas A. coffeaeformis and O. aurita were able to acclimate to UVR stress. These two species used different strategies. In A. coffeaeformis, the repair constant was lower than the damage constant, but phi(PSII) values returned to baseline values at the beginning of each experimental day, indicating that an effective active recovery process occurred after stress. In O. aurita, the repair processes took place during the stress, and could account for the UVR tolerance of this species.