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The effect of anoxia on the energy economy of root cells was studied by measuring heat production, ethanol and ATP production, K(+) fluxes and electrical activity in two Vitis species, V. riparia and V. rupestris, that differ in their tolerance to anoxia. Anoxia triggered a marked decrease of metabolic activity (measured by microcalorimetry) and of ATP levels in both species. In V. riparia after the first 2 h of anoxia, the decrease in the rate of heat production was not associated with a further significant decrease in ATP content, whereas in V. rupestris the ATP level continued to decrease until very low values were reached. The concomitant increase in the rate of ethanol production did not compensate for the decreased aerobic ATP supply. In V. rupestris, anoxia typically led to energy deficit and ATP imbalance, together with the subsequent disruption of ion homeostasis and cell death. In V. riparia, the strong decrease in K(+) membrane permeability together with the fast down-regulation of the electrical signals allowed the cells to avoid severe ion imbalances during prolonged anoxic episodes.

1 Hz rTMS over the supplementary motor area appeared to be effective in approximately 2/3 of SSRI-refractory OCD subjects, whereas in the TAU group only 1/4 of subjects were responders. The supplementary motor area might be a new target area to be further explored with neuromodulation for OCD treatment.

A simple procedure is described for the fabrication of micrometer to nanometer-scale platinum electrodes to be used in a vibrating oxygen-selective system. The electrode was prepared by etching a fine platinum wire and insulating it with an electrophoretic paint. The dimensions allowed this electrode to be used with the "vibrating probe technique" in exploratory studies aimed at mapping and measuring the patterns of net influxes as well as effluxes of oxygen in Olea europaea L. leaves and roots with spatial and temporal resolutions of a few microns and a few seconds, respectively. The magnitude and spatial localisation of O2 influxes in roots was characterised by two distinct peaks. The first, in the division zone, averaged 38 +/- 5 nmol m(-2) s(-1); the second, in the elongation region, averaged 68 +/- 6 nmol m(-2) s(-1). Long-term records of oxygen influx in the elongation region of the root showed an oscillatory regime characterised by a fast oscillation with periods of about 8-9 min. In leaves, the system allowed the measurement of real-time changes in O2 evolution following changes in light. Furthermore, it was possible to obtain "topographical" images of the photosynthetically generated oxygen diffusing through different stomata from a region of the leaf of 120 microm x 120 microm. The combination of topographic and electrochemical information at the micrometer scale makes the system an efficient tool for studying biological phenomena involving oxygen diffusion.

The effect of a hypoxic pre-treatment (HPT) on improving tolerance to prolonged anoxia conditions in two contrasting Vitis species (V. riparia, anoxia tolerant; V. rupestris, anoxia sensitive) was evaluated. The energy economy of root cells was studied by measuring heat production, the activity of pyruvate decarboxylase (PDC) and alcohol dehdrogenase (ADH), ethanol and ATP production, and K(+) fluxes. The results showed that HPT is an effective tool in order to maintain a sustainable metabolic performance in both the species under anoxia conditions, especially in sensitive species such as V. rupestris. Our results showed that the improved tolerance was mainly driven by: (i) an enhanced activity of key enzymes in alcohol fermentation (ADC and PDC); (ii) the capability to maintain a higher level of respiration, evidenced by a lesser decrease in heat development and ATP production; and (iii) the maintenance of a better ion homeostasis (highlighted by measurement of K(+) fluxes) and K(+) channel functionality.

Gaseous transport through lenticels is widely accepted to be the main pathway for oxygen supply to the parenchymatous tissues of the wood. Circumstantial evidence exists that the oxygen required for respiration by these living cells can be obtained from the transpiration stream. However, there has been no functional confirmation of this role. To address this problem and to quantify the contribution of the different pathways to the oxygen supply of the sapwood, we have developed a three-electrode miniaturized oxygen-selective sensor to be implanted into the sapwood for long-term determination of the oxygen concentration. In spring, during the active growing season, the oxygen concentration of the sapwood of young olive (Olea europaea L.) trees changed from 80-90 lmol O 2 l )1 around midday to 20-30 lmol O 2 l )1 in the night. These concentrations correspond to a deficit of oxygen for the sapwood between 65-70% and 88-90% of an aqueous solution saturated with air. In the daylight hours, almost all the oxygen present in the sapwood was delivered by the transpiration stream, driven by the soil-plant-atmosphere water-potential gradient. During the night the diffusion of oxygen via the sap-filled lumina of the tracheids and vessels (xylary diffusion in the aqueous phase) accounted for about 87% of all the oxygen present, whereas only the remaining 13% was assessed as supplied by radial diffusion in the aqueous or gaseous phase.

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Microemulsion EKC (MEEKC) was used for the determination of ketorolac and its three impurities. The microemulsion system was optimized, for the first time in the literature, using a multivariate strategy involving a mixture design. A 13-run experimental plan covering an experimental domain defined by the components aqueous phase (10 mM borate buffer pH 9.2), oil phase (n-heptane) and surfactant/cosurfactant (SDS/n-butanol) was carried out. Good results were obtained with all microemulsions tested considering as responses analysis time and resolution, and according to the desirability function the best microemulsion system was constituted by 90.0% 10 mM borate buffer, 2.0% n-heptane, 8.0% of SDS/n-butanol in 1:2 ratio. Finally, with the aim of reducing analysis time, a response surface study was carried out in the experimental domain defined by the process variables temperature and voltage and the best values were 17 degrees C and -17 kV, respectively. Applying the optimised conditions, a complete resolution among the analytes was obtained in about 3 min using the short-end injection method. The method was validated for both drug substances and drug product and was applied to the quality control of ketorolac in coated tablets. A comparison of MEEKC, MEKC and CEC for assaying ketorolac and its related substances has been made.