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Bracchitta G.,University of Catania | Bracchitta G.,Institute Curie | Bracchitta G.,French National Center for Scientific Research | Catalfo A.,University of Catania | And 8 more authors.
Photochemical and Photobiological Sciences | Year: 2013

Non-steroidal anti-inflammatory drugs (NSAID) are widely used in the treatment of pain and inflammation associated with several diseases. Naproxen, 2-(6-methoxy-2-naphthyl) propionic acid (NAP), belongs to this pharmacological class and appears to be associated with a high incidence of both photoallergic and phototoxic reactions. In this study, using human fibroblasts, we examined the biological effects of NAP photosensitization induced by UVA, the predominant UV component of sunlight reaching the Earth's surface. We showed that NAP or UVA alone have no cytotoxic effects at the concentrations and doses used in this study. The same result was observed when cells were pre-incubated with NAP but irradiated without NAP. In marked contrast, exposure of cells in the presence of NAP led to a drastic reduction of cell viability. These results suggest that the phototoxicity is mainly due to irradiation of extracellular NAP that damages cell membranes. Moreover, we showed that NAP itself led to a low but reproducible production of reactive oxygen species (ROS), to protein modifications by lipid peroxidation-derived aldehydes, to p38 phosphorylation and to the slowing-down of DNA replication, while UVA treatment alone showed no effects. NAP photosensitization with UVA led to protein S-glutathionylation, oxidation of the proliferating cell nuclear antigen (PCNA), oxidation of cellular tryptophan, phosphorylation of Chk1 and inhibition of DNA replication. However, using small interfering RNA to down regulate Chk1 expression in cells, we showed that Chk1 is not required to slow the S-phase down. Nevertheless, inhibition of Chk1, but not of p38, sensitized the cells to the phototoxic effects of NAP. Collectively, our data suggest that the interaction of NAP with the cells triggers oxidative damage and a replication stress, which are exacerbated by UVA radiation. As oxidative and replication stress-induced genome instability are important factors in aging and tumor predisposition, it is of interest to evaluate the consequence of a non-steroidal anti-inflammatory drug, like naproxen, on genomic instability. © 2013 The Royal Society of Chemistry and Owner Societies. Source


Bracchitta G.,University of Catania | Catalfo A.,University of Catania | De Guidi G.,University of Catania | De Guidi G.,Consorzio Interuniversitario Chimica per lAmbiente
Photochemical and Photobiological Sciences | Year: 2012

HPLC and emission spectroscopy were used to investigate UVA photosensitization of methylene blue (MB) or naproxen (NAP) towards bovine serum albumin (BSA). In addition, time resolved singlet oxygen measurements were carried out. The most stable drug : protein adducts stoichiometry of MB-BSA (1 : 1) and NAP-BSA (9 : 1) were verified by means of binding constant determination. UVA photosensitization of MB or NAP on BSA was studied by monitoring tryptophan (Trp) residue integrity. The sensitized photodegradation of the BSA resulted in different degrees of Trp damage. Thus, protein damage was determined by quantitative measurements of the different Trp (photo)-products. Indeed, many of these Trp derivatives are diagnostic for the photosensitization mechanism and some of them, for the first time in this work, were obtained by UVA photosensitization in proteins. The analysis of quantum yields of the photoproduct distribution allowed to weigh up the type I/II contribution to the UVA photosensitization mechanism. As expected, additional experiments in deuterated solvent resulted in an increase of the photodegradation quantum yields for those species where a singlet oxygen mechanism was involved. The UVA mediated generation of these Trp derivatives is consistent with the occurrence of singlet oxygen formation (almost dominant in MB), and photoionization (significant in NAP) within the protein matrix. Additional experiments at lower NAP concentration, as well as with human serum albumin (which differs for Trp content and, partially, localization), support further the molecular mechanism of photosensitization proposed. The results obtained in the case of this more complex system are in agreement with the free Trp model, even if, in almost all cases, the Trp photoproduct formation quantum yields are lower, due to the higher number of sensitization targets in the proteins. This journal is © The Royal Society of Chemistry and Owner Societies 2012. Source


Facchin V.,University of Verona | Cavinato C.,University of Venice | Pavan P.,University of Venice | Bolzonella D.,University of Verona | Bolzonella D.,Consorzio Interuniversitario Chimica per lAmbiente
Chemical Engineering Transactions | Year: 2013

Trace metals are essential for the enzyme cofactors involved in the biochemistry of methane formation and are needed in a balanced anaerobic digestion process. Food and kitchen waste generally contains low concentrations of trace elements, especially metals. As a consequence the anaerobic digestion process may result instable. The aim of this study is to evaluate the effect of metals addition on mesophilic anaerobic digestion of food waste, both in batch tests and in laboratory scale CSTR reactors. Batch anaerobic trials using source-separated food waste as substrate with inoculums of different origins were carried out under mesophilic conditions. Reactions were operated both with and without trace elements (Co, Mo, Ni, Se, W) supplementation. Supplementation with trace metals had either neutral or slightly negative effects with inoculums originating from reactors with a high background level of metals, such as those for the co-digestion of biowaste and waste activated sludge. For inoculums from reactors treating food waste only, which inherently contain low levels of trace metals, supplementation with these metals increased methane production. In particular, Mo concentrations in the range of 3-12 mg/kgTSfed and Se concentrations of 10 mg/kgTSfed increased methane production to as high as 30-40 %. Supplementation with a metal mixture (Co, Mo, Ni, Se, W) increased the methane production to the range 45-65 % for inoculums with low background concentrations of trace metals. These findings demonstrate the importance of Co, Mo, Ni, Se, W for high-performance anaerobic digestion process. Trace metals additions that showed the best batch results (100mgNi/kgTSfed, 100mgCo/kgTSfed, 6mgMo/kgTSfed, 10mgSe/kgTSfed, 10mgW/kgTSfed) were selected for CSTR experimentation. Performances of a continuous anaerobic digester fed with trace elements were compared to a "control reactor" fed with the same substrate but without metals addition. Preliminary CSTR results showed that metals addition allowed for a stable anaerobic digestion process at Organic Loading Rate greater than 3 kgVSfed m3•d but were not essential. Copyright © 2013, AIDIC Servizi S.r.l. Source


Passananti M.,Institute of Chemistry of Clermont-Ferrand | Passananti M.,CNRS Institute of Chemistry | Passananti M.,University of Naples Federico II | Temussi F.,University of Naples Federico II | And 7 more authors.
Water Research | Year: 2013

In this paper we investigated the degradation of the rivastigmine drug induced by hydroxyl radical in synthetic and natural waters focusing on both reactivity and photoproducts identification. The hydroxyl radical formation rate was quantified by using terephthalic acid as trapping molecule and it was related with the rivastigmine degradation rate. The second order rate constant between hydroxyl radical and rivastigmine was estimated to be ~5.8×109M-1s-1. Irradiation of rivastigmine in three natural waters (rain, lake and river) and comparison with degradation rates observed in synthetic solutions using nitrite, nitrate and hydrogen peroxide suggest that, in addition to hydroxyl radical, also nitroderived radicals (NO/NO2) are responsible for the pollutant degradation in natural media. In fact, the evaluated degradation rates in three natural waters are greatly higher than those estimated considering only the reactivity with photogenerated hydroxyl radical. Using nitrites and nitrates as photochemical OH source, the rivastigmine degradation cannot be described considering only the hydroxyl radical reactivity suggesting that NO and NO2 radicals could play a key role during indirect degradation. Moreover main degradation products have been identified by means of HPLC-MS. Hydroxylation of the aromatic ring as well as carbamate and amino chain oxidation were suggested as main reaction mechanisms, but also nitroderived compounds were characterized.Finally polychromatic irradiations of three rivastigmine doped natural waters (rain, river and lake) underlined the role of the indirect degradation that needs to be considered when direct degradation of selected pollutants is negligible under environmental-like conditions. © 2013 Elsevier Ltd. Source


Facchin V.,University of Verona | Cavinato C.,University of Venice | Fatone F.,University of Verona | Fatone F.,Consorzio Interuniversitario Chimica per lAmbiente | And 5 more authors.
Biochemical Engineering Journal | Year: 2013

Batch anaerobic trials using a source-separated food waste as a substrate with inoculums of different origins were carried out under mesophilic conditions. Reactions were operated both with and without trace element (Co, Mo, Ni, Se, and W) supplementation. Supplementation with trace metals had either neutral or slightly negative effects with inoculums originating from reactors with a high background level of metals, such as those for the co-digestion of biowaste and waste activated sludge. For inoculums from reactors treating food waste only, which inherently contain low levels of trace metals, supplementation with these metals increased methane production. In particular, Mo concentrations in the range of 3-12. mg/kg dry matter and Se concentrations of 10. mg/kg dry matter increased methane production to as high as 30-40%. Supplementation with a metal mixture (Co, Mo, Ni, Se and W) increased the methane production to the range 45-65% for inoculums with low background concentrations of trace metals. These findings may have an important impact in the commercial production of methane from food waste. © 2012 Elsevier B.V. Source

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