Biomedical subjects

C Gravato

Publications and source records attributed to C Gravato.

11 recordsLinked to original sources

PubMed2006

Oxidative stress, liver biotransformation and genotoxic effects induced by copper in Anguilla anguilla L.--the influence of pre-exposure to beta-naphthoflavone.

Fish are exposed in the aquatic ecosystems to different classes of pollutants. Polycyclic aromatic hydrocarbons (PAHs) and heavy metals represent two important classes of aquatic contaminants. Thus, one lot of European eels (Anguilla anguilla L.) was pre-exposed during 24 h to 2.7 microM beta-naphthoflavone (BNF; a PAH-like compound), and subsequently exposed during 24 h to 0, 1 and 2.5 microM copper (Cu). Additionally, another lot not pre-exposed to BNF was exposed to the same Cu concentrations. BNF pre-exposure promoted a significant increase in liver ethoxyresorufin O-deethylase (EROD) activity, but did not change the other responses investigated in eels. On the other hand, both Cu concentrations did not modify the liver EROD activity either in eels pre-exposed to BNF or not. Liver total cytochrome P450 was increased in eels exposed to Cu 2.5 microM, being significantly only in eels not pre-exposed to BNF. Free sulfhydryl group content was decreased by 1 and 2.5 microM in eels pre-exposed to BNF or not pre-exposed, being significant at 2.5 microM Cu in eels not pre-exposed compared to its control. Liver total glutathione (TG), reduced glutathione (GSH) and GSH/oxidized glutathione (GSSG) levels were slightly decreased by 1 and 2.5 microM Cu in eels pre-exposed to BNF, whereas a slight tendency to increase was observed in eels not pre-exposed. Thus, liver TG and GSH significantly decreased in 2.5 microM Cu BNF pre-exposed eels compared to eels not pre-exposed to BNF. Liver glutathione reductase and catalase activities were significantly inhibited by 1 and 2.5 microM Cu in eels pre-exposed to BNF, concomitantly with a slight liver glutathione peroxidase tendency to decrease. Lipid peroxidation was significantly increased by 1 microM Cu in eels either pre-exposed or not pre-exposed to BNF. Liver H(2)O(2) was significantly increased by 1 microM Cu in eels pre-exposed to BNF. Liver DNA integrity was significantly decreased by 1 and 2.5 microM Cu in eels pre-exposed to BNF. The oxidative stress and genotoxic effects induced by Cu in eels pre-exposed to BNF revealed that the metal effects are potentiated by previous exposure to BNF.

Anguilla↗

PubMed2005

Oxidative stress and genotoxic responses to resin acids in Mediterranean mussels.

This study represents the first attempt to investigate the genotoxic effects and oxidative stress of resin acids in Mediterranean mussels (Mytilus galloprovincialis Lmk). Mussels were exposed to 2.7 microM abietic acid (AA) and dehydroabietic acid (DHAA) for 6, 12, 18, and 24h. Gill and hepatopancreas conjugation activity, antioxidant defense system, lipid peroxidation (LPO), and DNA damage were determined as reduced glutathione (GSH), glutathione S-transferase (GST) activity, glutathione peroxidase (GPx) activity, catalase (CAT) activity, LPO, and DNA strand breaks. AA caused significant GST inhibition in mussel gills at 12, 18, and 24h. Activity of the antioxidant enzymes, namely, GPx and CAT, was inhibited at 24 and 18 h, respectively, in mussel gills. A significant increase in gill LPO was observed at 24h. The DNA integrity of mussel hepatopancreas significantly decreased after 12 and 24 h exposure to AA. A significant increase in LPO was observed after 6h exposure to DHAA, in either mussel gills or hepatopancreas. DNA integrity was significantly decreased in mussel hepatopancreas after 12 and 24 h exposure to DHAA. AA induced oxidative damage and genotoxicity in mussels, because it promoted increases in LPO in gills and DNA strand breaks in hepatopancreas. DHAA promoted oxidative damage and genotoxicity in mussels, as significant increases were observed in LPO in gills and hepatopancreas and in DNA strand breaks in hepatopancreas.

Abietanes↗

PubMed2004

Juvenile sea bass biotransformation, genotoxic and endocrine responses to beta-naphthoflavone, 4-nonylphenol and 17 beta-estradiol individual and combined exposures.

Juvenile sea bass, Dicentrarchus labrax L., were exposed during 2, 4, 8, and 24 h to 0.9 microM beta-naphthoflavone (BNF), 131 nM 17 beta-estradiol (E(2)), 4.05 microM 4-nonylphenol (NP), as well as to BNF combined either to E(2) or NP (maintaining the previous concentrations). Liver cytochrome P450 content (P450), ethoxyresorufin-O-deethylase (EROD), and glutathione S-transferase (GST) activities were measured in order to evaluate biotransformation responses. Genotoxicity was assessed as erythrocytic nuclear abnormalities (ENA) frequency. The effects on endocrine function were evaluated as plasma cortisol and glucose. Cortisol was not affected by xeno/estrogens tested, either in single exposure or mixed with BNF. Nevertheless, the intermediary metabolism was affected since glucose concentration increased after 4 h exposure to E(2), and after all BNF+NP exposure lengths. Moreover, a synergism between BNF and NP was thoroughly demonstrated, whereas a sporadic antagonistic interaction was found at 4 h BNF + E(2) exposure. Liver EROD and GST activities were not significantly altered by single E(2) or NP exposure. However, both compounds were able to induce EROD activity in the presence of BNF. NP single exposure was able to significantly increase liver P450 content, while its mixture with BNF displayed an antagonistic interference. Considering the xeno/estrogens single exposures, only NP induced an ENA increase; however, both mixtures (BNF + E(2) and BNF + NP) displayed genotoxic effects. Fish responses to mixtures of xenobiotics are complex and the type of interaction (synergism/potentiation or antagonism) in a particular mixture can vary with the evaluated biological response.

Animals↗

PubMed2003

Genotoxicity biomarkers' association with B(a)P biotransformation in Dicentrarchus labrax L.

Dicentrarchus labrax (sea bass) were exposed during 0, 4, 8, 16, 24, 48, and 96 h to 0 and 0.1 microM benzo(a)pyrene (B(a)P), an environmental pollutant, and the following biomarkers were measured: (1) liver cytochrome P450 (P450) content and ethoxyresorufin-O-deethylase (EROD) activity as phase I biotransformation parameters, (2) liver gluthathione-S-transferase (GST) activity as a phase II biotransformation conjugation enzyme, (3) biliary and liver cytosolic B(a)P-type metabolites by fixed wavelength fluorescence detection (FF), and (4) erythrocytic micronuclei (EMN) and erythrocytic nuclear abnormalities (ENA) as genotoxicity biomarkers. Liver EROD activity (4 h), P450 content (24 h), GST activity (4, 8, and 96 h), bile (4-96 h), and liver cytosolic (4-24 h) B(a)P-type metabolites increased significantly in sea bass exposed to B(a)P as well as EMN (8-96 h) and ENA (4-96 h) frequencies. B(a)P genotoxicity is associated with increase in B(a)P-type metabolites in liver cytosol due to an impaired phase II conjugation. This increase seems to be responsible for the decrease in liver EROD and GST activities.

Animals↗

PubMed2003

Dicentrarchus labrax biotransformation and genotoxicity responses after exposure to a secondary treated industrial/urban effluent.

The present research work was designed to study Dicentrarchus labrax (sea bass) biotransformation and genotoxicity responses to the soluble fraction of a secondary treated industrial/urban effluent (SF-STIUE) discharged through a submarine pipe outlet into the Aveiro coastal area. Sea bass was exposed for 4, 8, 16, 24, 48, and 96 h to 0%, 0.1%, and 1% SF-STIUE and the following biological responses were measured: (1) liver cytochrome P450 (P450) content and ethoxyresorufin-O-deethylase (EROD) activity, as phase I biotransformation parameters; (2) liver gluthathione S-transferase (GST) activity as a phase II conjugation enzyme; (3) biliary and liver cytosol naphthalene (Naph)- and benzo(a)pyrene (B(a)P)-type metabolites, by fixed wavelength fluorescence detection (FF); (4) liver DNA strand breaks, erythrocytic micronuclei (EMN), and erythrocytic nuclear abnormalities (ENA) as genotoxicity parameters. Both SF-STIUE dilutions (0.1% and 1%) failed to significantly increase liver EROD activity, despite a significant increase of liver P450 at 16 and 48 h exposure to 0.1%. Liver GST activity increased significantly at 4h of sea bass exposure to 1% SF-STIUE, being inhibited at 96 h of exposure to this SF-STIUE dilution. Naph- and B(a)P-type metabolite contents were not significantly increased in bile. However, Naph-type metabolite contents increased significantly in liver cytosol at 4h exposure to 1% SF-STIUE, and at 24h exposure to 0.1% and 1% SF-STIUE. Furthermore, B(a)P-type metabolites increased significantly in liver cytosol at 4h exposure to 1% SF-STIUE, and 16 h exposure to 0.1% and 1% SF-STIUE. EMN and ENA frequencies increased significantly at 4, 8, 16, 24, 48, and 96 h exposure to 0.1% and 1% SF-STIUE. Liver DNA integrity decreased significantly at 96 h of sea bass exposure to 1% SF-STIUE. The STIUE discharged into Aveiro coastal area is of great ecotoxicological concern due to its genotoxic potential.

Animals↗

PubMed2002

Juvenile sea bass liver P450, EROD induction, and erythrocytic genotoxic responses to PAH and PAH-like compounds.

Sea bass were exposed, for 0, 2, 4, 6 and 8 h, to 0 or 2.7 microM beta-naphthoflavone (BNF), or to 0, 0.1, 0.3, 0.9, or 2.7 microM benzo[a]pyrene (B(a)P) and naphthalene (NAPH). Liver cytochrome P-450 content (P450) and liver ethoxyresorufin-O-deethylase activity (EROD) induction were determined as phase I biotransformation responses, whereas erythrocytic micronuclei (EMN) and erythrocytic nuclear abnormalities (ENA) tests were performed to assess genotoxic effects. Liver alanine aminotransferase activity and liver somatic index were determined, respectively, as liver damage and common health indicators. A significant increase in sea bass EROD activity began, respectively, at 2 and 4 h exposure to 2.7 microM B(a)P and 2.7 microM BNF, whereas NAPH failed to induce EROD activity. Maximal EROD activity was observed after 6 h exposure to 2.7 microM BNF (9-fold increase), 2.7 microM B(a)P (4-fold increase), and 2.7 microM NAPH (2-fold increase), indicating BNF as the most potent EROD inducer (BNF>B(a)P>NAPH). A significant increase in liver P450 content was observed at 6 h exposure to 2.7 microM BNF (6.5-fold increase), indicating BNF as the most potent P450 inducer. A significant P450 increase was observed at 8 h exposure only to 0.1 microM B(a)P (2-fold increase), whereas it slightly increased at 2 h exposure to 2.7 microM NAPH, within a wide variable range. The BNF, NAPH, and B(a)P genotoxic potential was demonstrated as sea bass EMN and ENA. B(a)P promoted at 2 h exposure a significant EMN (24-fold) and ENA (2.2-fold) increase, whereas NAPH exhibited similar results only at 8 h exposure. BNF also increased significantly sea bass EMN (8-fold) and ENA (1.5-fold) after 8 h. The results indicated B(a)P as the most genotoxic compound, followed by NAPH and BNF (B(a)P>NAPH>BNF). Despite the low liver P450 content and EROD activity induction by NAPH and B(a)P, their genotoxic potential was higher than that of BNF.

Alanine Transaminase↗

PubMed2002

Liver phase I and phase II enzymatic induction and genotoxic responses of beta-naphthoflavone water-exposed sea bass.

Sea bass were exposed to 0 (control), 0.1, 0.3, 0.9, and 2.7 microM beta-naphthoflavone (BNF) for 0, 2, 8, and 16 h in order to assess the chronological and concentration relationships between BNF phase I and II biotransformation responses, such as liver cytochrome P450 (P450) content, ethoxyresorufin-O-deethylase (EROD), uridine diphosphate-glucuronosyl transferase (UDP-GT), and the genotoxic effects, measured either by erythrocytic micronuclei (EMN) or erythrocytic nuclear abnormalities (ENA) tests. Liver alanine aminotransferase (ALT) activity and liver somatic index (LSI) were also measured. A significant liver EROD activity was found at 8 h exposure, respectively, to 0.1, 0.3, 0.9, and 2.7 microM BNF. Maximal liver EROD activity increase was observed at 16 h exposure to 0.9 microM BNF, whereas the highest liver P450 was reached at 8 h exposure to 2.7 microM BNF. Liver UDP-GT activity was significantly increased at 2 h exposure to 0.1 and 0.3 microM BNF and at 8 h exposure to 0.1, 0.3, and 0.9 microM BNF, decreasing at 16 h, for every exposure concentration. Significant ENA increase was observed at 2h exposure, respectively, to 0.3, 0.9, and 2.7 microM BNF. Maximal ENA increase was observed at 16 h exposure to 0.9 microM BNF. The MN was significantly increased at 8 and 16 h exposure, respectively, to 2.7 and 0.9 microM BNF. Liver ALT activity significantly increases at 8 h exposure to 0.1 and 0.3 microM BNF, whereas liver somatic index was significantly increased from 2 to 16 h exposure for every BNF concentration. A slight liver EROD activity increase with a concomitant lack of liver UDP-GT activity is able to induce significant erythrocytic genotoxic effects. Liver UDP-GT high levels are important in sea bass BNF detoxification. However, high liver UDP-GT activity is not enough to prevent the BNF metabolite genotoxic effects on sea bass erythrocytes when liver EROD activity is induced at 2 and 8 h exposure to 0.3 and 0.9 microM BNF. The genotoxic effects measured as EMN and ENA suggest that the balance between the rates of liver BNF reactive and conjugated metabolites seems to be critical.

Animals↗

PubMed2002

Beta-naphthoflavone liver EROD and erythrocytic nuclear abnormality induction in juvenile Dicentrarchus labrax L.

Juvenile Dicentrarchus labrax L. (sea bass) was exposed to five different beta-naphthoflavone (BNF) concentrations-0, 0.1, 0.3, 0.9 and 2.7 microM-for 0, 2, 4, 6, 8, 16, 24, 48, 72, 144, and 216 h. A battery of biomarkers was investigated, such as liver ethoxyresorufin-O-deethylase (EROD), liver cytochrome P450 (P450 content), liver aminotransferase (ALT activity), liver somatic index (LSI), micronuclei (MN), and erythrocytic nuclear abnormality (ENA) frequencies. Juvenile D. labrax L. liver EROD induction started at 2 h exposure to 2.7 microM BNF and 6 h exposure to 0.1, 0.3, and 0.9 microM BNF, respectively. A significant liver EROD decrease was observed between 8 and 16 h for all BNF concentrations, followed by a slight increase after 48 h exposure to 0.9 and 2.7 microM BNF and a definitive decrease from 72 h exposure onward. Liver P450 content significantly increased at 2, 6, and 8 h exposure, respectively, to 2.7 microM, 0.9, 0.3, and 0.1 microM BNF. However, liver P450 content remained significantly higher than that of the control from 72 to 216 h in the sea bass exposed to 2.7 microM BNF. Sea bass ENA induction started at 4h exposure to 0.9 and 2.7 microM BNF, and significantly increased to 16 and 24 h exposure, whereas for 0.3 microM BNF a significant increase started after 8 h exposure. A significant ENA frequency increase was still observed at 144 and 216 h exposure to 0.9 and 2.7 microM BNF. The micronuclei induction was observed at 4, 6, and 8 h, respectively, after 2.7, 0.9, and 0.3 microM BNF exposure. However, there was a micronuclei frequency decrease for 0.3, 0.9, and 2.7 microM BNF exposure concentrations between 8 and 16 h, followed by a slight increase after 48, 72, and 144 h exposure, respectively, to 2.7, 0.9, and 0.3 microM BNF. Liver somatic index significantly increased after 216 h, whereas ALT activity significantly decreased at 144 and 216 h 2.7 microM BNF exposure.

Animals↗

PubMed2002

Juvenile sea bass liver biotransformation induction and erythrocytic genotoxic responses to resin acids.

Juvenile Dicentrarchus labrax L. (sea bass) were exposed for 0, 2, 4, 6, and 8 h to abietic acid (AA) and dehydroabietic acid (DHAA) in concentration ranges 0, 0.0125, 0.025, 0.05, 0.1, 0.3, 0.9, and 2.7 microM. Liver cytochrome P-450 content (P450) and liver ethoxyresorufin-O-deethylase activity (EROD) were determined as phase I biotransformation biomarkers. Genotoxicity was measured as erythrocytic micronuclei (EMN) and nuclear abnormalities (ENA). Liver damage was assessed as liver alanine aminotransferase activity (ALT) and liver somatic index (LSI) was used as a general health indicator. AA inhibited EROD (at 2 h exposure to 0.05 microM) or failed to induce it, whereas a significant P450 increase was observed at 2 h exposure to 0.05 (2.3-fold) and 2.7 microM (6.3-fold). A significant P450 increase was also observed at 6 and 8 h exposure, respectively, to 0.0125 (3.4-fold) and 0.025 microM (4.9-fold) AA. EMN and ENA frequency were significantly increased at 2 h exposure to 0.9 microM AA. A significant EMN and ENA time-related increase was observed with an increased exposure length up to 8 h. Therefore, all the AA concentrations tested promoted an EMN and ENA increase at 8 h exposure. DHAA induced a significant EROD increase (3.2-fold) at 2 h exposure to the lowest concentration tested. Liver P450 was significantly increased at 2 h exposure to 0.025 (1.8-fold) and 0.3 microM (2.5-fold), at 4 h exposure to 0.1 microM (3.6-fold), and at 6 h exposure to 0.1 (3.2-fold) and 0.3 microM (2.8-fold), whereas it was significantly decreased (30% of control value) at 4 h exposure to 0.9 microM. EMN and ENA frequency were significantly increased from 0 to 2 h exposure to all DHAA concentrations tested and remained high from 2 to 8 h. EMN frequency was increased 20 times over control at 2 h exposure to 0.0125 microM DHAA, whereas it increased 9-fold at 2 h exposure to 0.9 microM AA. Furthermore, ENA frequency was increased 3 times over control at 2 h exposure to 0.0125 microM DHAA, and 2-fold after exposure to 0.9 microM AA. Maximal EMN and ENA induction was observed at 2 h exposure to 0.0125 microM DHAA and only at 4 h exposure to 2.7 microM AA. Therefore, the comparative analysis of the two resin acids genotoxic effects, measured as EMN and ENA, indicated that DHAA is more genotoxic than AA.

Abietanes↗

PubMed2002

Juvenile sea bass liver biotransformation and erythrocytic genotoxic responses to pulp mill contaminants.

This research represents the first study concerning liver phase I biotransformation induction, measured as cytochrome P-450 (P450) and ethoxyresorufin-O-deethylase (EROD), and genotoxic responses, measured as erythrocytic micronuclei (EMN) and nuclear abnormalities (ENA), in Dicentrarchus labrax L. (sea bass) exposed to pulp and paper mill contaminants. Juvenile sea bass were exposed for 6 h to 0 (control) and 0.0125 microM concentrations of the resin acids (RAs) abietic acid (AA) and dehydroabietic acid (DHAA) or 7-isopropyl-1-methylphenanthrene (retene) (Experiment 1). Sea bass were exposed for 6 h to 0 (control), 0.78, 1.56, 3.12, 6.25, and 12.5% bleached kraft pulp mill environmental contaminated water (BKPMECW) collected at the old sewage outlet of a pulp and paper industry (Experiment 2). The time-dependent response was studied in sea bass at 0, 8, 16, 24, 48, and 72 h exposure to 0.78 and 12.5% BKPMECW (Experiment 3). The experimental results demonstrated the presence of genotoxic compounds in BKPMECW. AA, DHAA, and retene may be the constituents responsible for high BKPMECW genotoxicity, since they induced similar sea bass EMN and ENA frequency increases. The BKPMECW, the RAs, and retene failed to significantly increase liver EROD activity and P450 content at 6 h. Furthermore, 3.12% BKPMECW, 0.0125 microM AA, and 0.0125 microM retene significantly decreased liver EROD activity. However, P450 was significantly increased from 8 up to 72 h exposure to BKPMECW. Therefore, the low or inhibited EROD levels could be a consequence of a general membrane disturbance by BKPMECW, RAs, and retene. However, liver ALT results indicate significant liver damage or enzyme inhibition only at 8, 16, 48, and 72 h exposure to BKPMECW.

Animals↗

PubMed1998

Ca2+-H+ antiport activity in synaptic vesicles isolated from sheep brain cortex.

Synaptic vesicles isolated from sheep brain cortex exhibit an ATP-dependent Ca2+ accumulation that is inhibited by the protonophore uncoupler carbonyl cyanide m-chorophenylhydrazone (CCCP) and completely released by the Ca2+ ionophore ionomycin. This transport activity was sensitive to the V-type ATPase inhibitor, bafilomycin, but not to the P-type ATPase inhibitor, vanadate. We also observed that the proton gradient, established across the synaptic vesicle membranes in the presence of ATP, is partially dissipated by the addition of Ca2+ (100-860 microM) in correlation to an increase of ATP hydrolysis by the H+-pumping ATPase. In contrast, the activity of the H+-ATPase, measured under uncoupling conditions (presence of CCCP), appears to be unaltered by the calcium ion. The Ca2+-induced H+ release visualized by fluorescence quenching of acridine orange correlates well with the Ca2+ uptake determined isotopically. These results indicate that synaptic vesicles accumulate Ca2+, via a low affinity Ca2+-H+ antiport system energized by the protonmotive force originated from the H+-pumping ATPase activity.

Adenosine Triphosphate↗