Ester Marotta

A mitochondriotropic derivative of quercetin: a strategy to increase the effectiveness of polyphenols.

Mitochondria‐targeted compounds are needed to act on a variety of processes that take place in these subcellular organelles and that have great pathophysiological relevance. In particular, redox‐active molecules that are capable of homing in on mitochondria provide a tool to intervene on a major cellular source of reactive oxygen species and on the processes they induce, notably the mitochondrial permeability transition and cell death. We have linked the 3‐OH of quercetin (3,3′,4′,5,7‐pentahydroxy flavone), a model polyphenol, and the triphenylphosphonium moiety, a membrane‐permeant cationic group, to produce proof‐of‐principle mitochondriotropic quercetin derivatives. The remaining hydroxyls were sometimes acetylated to hinder metabolism and improve solubility. The new compounds accumulate in mitochondria in a transmembrane potential‐driven process and are only slowly metabolised by cultured human colon cells. They inhibit mitochondrial ATPase activity much as quercetin does, and are toxic for fast‐growing cells.

Atmospheric pressure photoionization mechanisms - 1. The case of acetonitrile

Abstract Experimental data show that acetonitrile, even though it has an ionization energy higher than the photon energy employed in atmospheric pressure photoionization (APPI) conditions, participates in the protonation of furocumarins. In order to clarify this unexpected behaviour, the processes activated by APPI on acetonitrile have been studied in detail. The formation of protonated molecules (C2H4N+ cations), and of a complex between acetonitrile and water (C2H6NO+ cations), is observed. To clarify the nature of this species, 13 C and 2 H labelling experiments and product ion spectra have been employed. The data suggest that photon irradiation leads to a first isomerization of acetonitrile molecules that leads to species that exhibit an IE

Biodegradation of Chlorsulfuron and Metsulfuron‐Methyl by Aspergillus niger in Laboratory Conditions

Two sulfonylurea herbicides, chlorsulfuron and metsulfuron‐methyl, were studied under laboratory conditions, in order to elucidate the biodegradation pathway operated by Aspergillus niger, a common soil fungus, which is often involved in the degradation of xenobiotics. HPLC‐UV was used to study the kinetic of degradation, whereas LC‐MS was used to identify the metabolites structure. In order to avoid the chemical degradation induced by a decrease in pH, due to the production of citric acid by the fungus, the experiments were performed in a buffered neutral medium. No significant degradation for both compounds was observed in mineral medium with 0.2% sodium acetate. On the contrary, in a rich medium, after 28 days the degradations, chemical degradation excluded, were about 30% for chlorsulfuron and 33% for metsulfuron‐methyl. The main microbial metabolites were obtained via cleavage of the sulfonylurea bridge. In addition the fungus seems to be able to hydroxylate the aromatic ring of chlorsulfuron. In the case of metsulfuron‐methyl the only detected metabolite was the triazine derivative, while the aromatic portion was completely degraded. Finally, the demethylation of the methoxy group on the triazine ring, previously observed with a Pseudomonas fluorescens strain, was not observed with A. niger.

Comparison of Toluene Removal in Air at Atmospheric Conditions by Different Corona Discharges

Different types of corona discharges, produced by DC of either polarity (+/-DC) and positive pulsed (+pulsed) high voltages, were applied to the removal of toluene via oxidation in air at room temperature and atmospheric pressure. Mechanistic insight was obtained through comparison of the three different corona regimes with regard to process efficiency, products, response to the presence of humidity and, for DC coronas, current/voltage characteristics coupled with ion analysis. Process efficiency increases in the order +DC < -DC < +pulsed, with pulsed processing being remarkably efficient compared to recently reported data for related systems. With -DC, high toluene conversion and product selectivity were achieved, CO(2) and CO accounting for about 90% of all reacted carbon. Ion analysis, performed by APCI-MS (Atmospheric Pressure Chemical Ionization-Mass Spectrometry), provides a powerful rationale for interpreting current/voltage characteristics of DC coronas. All experimental findings are consistent with the proposal that in the case of +DC corona toluene oxidation is initiated by reactions with ions (O(2)(+*), H(3)O(+) and their hydrates, NO(+)) both in dry as well as in humid air. In contrast, with -DC no evidence is found for any significant reaction of toluene with negative ions. It is also concluded that in humid air OH radicals are involved in the initial stage of toluene oxidation induced both by -DC and +pulsed corona.

Impact of mitochondriotropic quercetin derivatives on mitochondria.

Mitochondria-targeted polyphenols are being developed with the intent to intervene on the levels of reactive oxygen species (ROS) in mitochondria. Polyphenols being more than just anti-oxidants, the interaction of these derivatives with the organelles needs to be characterised. We have studied the effects of two quercetin derivatives, 3-(4-O-triphenylphosphoniumbutyl)quercetin iodide (Q3BTPI) and its tetracetylated analogue (QTA3BTPI), on the inner membrane aspecific permeability, transmembrane voltage difference and respiration of isolated rat liver mitochondria. While the effects of low concentrations were too small to be reliably defined, when used in the 5-20 microM range these compounds acted as inducers of the mitochondrial permeability transition (MPT), an effect due to pro-oxidant activity. Furthermore, Q3BTPI behaved as an uncoupler of isolated mitochondria, causing depolarisation and stimulating oxygen consumption. When applied to tetramethylrhodamine methyl ester (TMRM)-loaded HepG2 or Jurkat cells uptake of the compounds was predictably associated with a loss of TMRM fluorescence, but there was no indication of MPT induction. A production of superoxide could be detected in some cells upon prolonged incubation of MitoSOX-loaded cells with QTA3BTPI. The overall effects of these model mitochondriotropic polyphenols may thus differ considerably depending on whether their hydroxyls are protected or not and on the experimental system. In vivo assays will be needed for a definitive assessment of their bioactivities.

Regioselective O-derivatization of quercetin via ester intermediates. An improved synthesis of rhamnetin and development of a new mitochondriotropic derivative.

The regioselective synthesis of several quercetin (3,3’,4’,5,7-pentahydroxy flavone) tetraesters bearing a single free OH on 5-C was achieved in good yield by proper choice of reaction conditions using common esterification procedures. Tetracetylated quercetin with the free OH on 7-C was selectively obtained instead via imidazole-promoted deacylation of the corresponding pentaester. Unambiguous structural characterization of the two isomeric tetraacetyl quercetin derivatives was obtained by combined HSQC and HMBC 2D-NMR analysis. These molecules can be used as starting materials for the regioselective synthesis of other derivatives. High yield syntheses of the natural polyphenol rhamnetin (7-O-methylquercetin) and of the new mitochondriotropic compound 7-(4-triphenylphosphoniumbutyl) quercetin iodide are reported as examples.

Comparison of the rates of phenol advanced oxidation in deionized and tap water within a dielectric barrier discharge reactor

Electric non-thermalizing discharges provide promising novel means to induce oxidation of organic pollutants in water. The decomposition of phenol in solutions prepared with deionized (milliQ) and tap water was studied and compared in a Dielectric Barrier Discharge (DBD) reactor. Interestingly, a significant rate increase was found in tap with respect to milliQ water. Control experiments proved that this was not the effect of conductivity or of traces of iron or of residual active chlorine from the depuration process operated in the aqueducts of Italian cities. The same increase in efficiency as observed in tap water was instead obtained when phenol was treated in solutions containing bicarbonate anions in the same concentration as present in tap water, an effect attributed to buffering of the solution pH. The role of pH has been investigated thoroughly by measuring the process efficiency over a wide pH range, from 2 to 10, by using different buffer systems to probe reactivity at near neutral pH, the most relevant for drinking water applications, and by testing the effect of different buffer concentrations. These latter experiments failed to detect any significant kinetic effect attributable to the well known reactivity of bicarbonate as quencher of OH radicals.

Effect of vegetative filter strips on herbicide runoff under various types of rainfall.

Narrow vegetative filter strips proved to effectively reduce herbicide runoff from cultivated fields mainly due to the ability of vegetation to delay surface runoff, promote infiltration and adsorb herbicides. A field trial was conducted from 2007 to 2009 in north-east Italy in order to evaluate the effectiveness of various types of vegetative filter strips to reduce spring-summer runoff of the herbicides mesotrione, metolachlor and terbuthylazine, widely used in maize, and to evaluate the effect of the rainfall characteristics on the runoff volume and concentration. Results show that without vegetative filter strip the herbicide load that reaches the surface water is about 5-6 g ha(-1)year(-1) for metolachlor and terbuthylazine (i.e. 0.5-0.9% of the applied rate), confirming that runoff from flat fields as in the Po Valley can have a minor effect on the water quality, and that most of the risk is posed by a few, or even just one extreme rainfall event with a return period of about 25-27 years, causing runoff with a maximum concentration of 64-77 μg L(-1). Mesotrione instead showed rapid soil disappearance and was observed at a concentration of 1.0-3.8 μg L(-1) only after one extreme (artificial) rainfall. Vegetative filter strips of any type are generally effective and can reduce herbicide runoff by 80-88%. Their effectiveness is steady even under severe rainfall conditions, and this supports their implementation in an environmental regulatory scheme at a catchment or regional scale.

Absorption and Metabolism of Resveratrol Carboxyesters and Methanesulfonate by Explanted Rat Intestinal Segments

Model prodrugs of resveratrol carrying protecting substituents at the hydroxyls have been synthesised and tested. Resveratrol triacetate and resveratrol-tri-mPEG1900 were formed by linking methyl groups or poly(ethylene glycol) chains, respectively, via carboxyester bonds. Resveratrol trimesylate, a molecule less susceptible to hydrolytic attack, was synthesised as well. This latter compound proved to be stable in vitro, while the carboxyester derivatives were slowly hydrolysed in solutions mimicking the gastric or intestinal environment, and rapidly converted to resveratrol in blood. In ex vivo permeation experiments with explanted intestinal segments, resveratrol and its triacetate derivative appeared in the basolateral compartment essentially as a mixture of Phase II metabolites. When the PEGylated derivative was provided on the apical side, unconjugated resveratrol accounted for about 50% of the compounds in the basolateral-side chamber. The same result was obtained by providing an equivalent physical mixture of resveratrol and PEG polymer, indicating that this behaviour is likely due to an adjuvating effect of PEG rather than to the covalent polymer conjugation. These observations suggest that the ester derivatives are rapidly hydrolysed at the intestinal surface or inside enterocytes, and are then processed as resveratrol. On the other hand, the mesylate was transported from the apical to the basolateral side without modification. It may thus be possible to enhance absorption and hinder metabolism of natural polyphenols by constructing pro-drugs incorporating bonds with appropriate resistance to enzymatic hydrolysis.

Oxidation mechanisms of CF2Br2 and CH2Br2 induced by air nonthermal plasma.

Oxidation mechanisms in air nonthermal plasma (NTP) at room temperature and atmospheric pressure were investigated in a corona reactor energized by +dc, -dc, or +pulsed high voltage.. The two bromomethanes CF(2)Br(2) and CH(2)Br(2) were chosen as model organic pollutants because of their very different reactivities with OH radicals. Thus, they served as useful mechanistic probes: they respond differently to the presence of humidity in the air and give different products. By FT-IR analysis of the postdischarge gas the following products were detected and quantified: CO(2) and CO in the case of CH(2)Br(2), CO(2) and F(2)C ═ O in the case of CF(2)Br(2). F(2)C ═ O is a long-lived oxidation intermediate due to its low reactivity with atmospheric radicals. It is however removed from the NTP processed gas by passage through a water scrubber resulting in hydrolysis to CO(2) and HF. Other noncarbon containing products of the discharge were also monitored by FT-IR analysis, including HNO(3) and N(2)O. Ozone, an important product of air NTP, was never detected in experiments with CF(2)Br(2) and CH(2)Br(2) because of the highly efficient ozone depleting cycles catalyzed by BrOx species formed from the bromomethanes. It is concluded that, regardless of the type of corona applied, CF(2)Br(2) reacts in air NTP via a common intermediate, the CF(2)Br radical. The possible reactions leading to this radical are discussed, including, for -dc activation, charge exchange with O(2)(-), a species detected by APCI mass spectrometry.