Environmental pollutants, such as dioxin-like (DL) PCBs, benzo(a) pyrene (B[a]P), and flavonoids ... more Environmental pollutants, such as dioxin-like (DL) PCBs, benzo(a) pyrene (B[a]P), and flavonoids are aryl hydrocarbon receptor (AHR) ligands and may be excreted in dairy milk. The expression of AHR-target genes, particularly those involved in xenobiotic biotransformation, and their modulation by two DL-PCBs, B[a]P, and β-naphthoflavone was investigated in a bovine mammary epithelial cell line (BME-UV). As assessed by quantitative PCR, BME-UV cells expressed a functional AHR signaling pathway. All the AHR ligands induced a concentration-related increase in the transcription of cytochrome P450 1A1 and 1B1, known to be implicated in the bioactivation of several xenobiotics. Conversely, genes encoding for antioxidant and detoxifying enzymes, like quinone oxidoreductase or glutathione S-transferase A2, were not affected or even depressed. This study demonstrates the occurrence and the modulation by different AHR-ligands of genes involved in xenobiotic metabolism in BME-UV cells, with the...
Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment, 2015
A chemometric class modelling strategy (unequal dispersed classes - UNEQ) was applied for the fir... more A chemometric class modelling strategy (unequal dispersed classes - UNEQ) was applied for the first time as a possible screening method to monitor the abuse of growth promoters in veal calves. Five serum biomarkers, known to reflect the exposure to classes of compounds illegally used as growth promoters, were determined from 50 untreated animals in order to design a model of controls, representing veal calves reared under good, safe and highly standardised breeding conditions. The class modelling was applied to 421 commercially bred veal calves to separate them into 'compliant' and 'non-compliant' with respect to the modelled controls. Part of the non-compliant animals underwent further histological and chemical examinations to confirm the presence of either alterations in target tissues or traces of illegal substances commonly administered for growth-promoting purposes. Overall, the congruence between the histological or chemical methods and the UNEQ non-compliant o...
7,12-Dimethylbenzanthracene (DMBA) is an abundant environmental contaminant, which undergoes bioa... more 7,12-Dimethylbenzanthracene (DMBA) is an abundant environmental contaminant, which undergoes bioactivation, primarily by the CYP1 family, both in liver and extra-hepatic tissues. Dietary acetylsalicylic acid (ASA) has been recently reported to inhibit DMBA-mediated mammary tumour formation in rats. Chemopreventive substances may reduce the risk of developing cancer by decreasing metabolic enzymes responsible for generating reactive species (phase I enzymes) and/or increasing phase II enzymes that can deactivate radicals and electrophiles. To test these hypotheses, Sprague-Dawley female rats were orally administered ASA as lysine acetylsalicylate (50 mg per capita/day for 21 days in water), DMBA (10 mg per capita in olive oil on day 7, 14, and 21), ASA and DMBA in combination, and vehicles only, respectively. Six rats for each group were sacrificed on day 8, 15, and 22. The DMBA-mediated increase in hepatic CYP1A expression and related activities was not significantly affected by ASA, which, conversely, enhanced in a time-dependent manner the liver reduced glutathione content (up to 52%) and the activity of NAD(P)H-quinone oxidoreductase (up to 34%) in DMBA-treated rats. It is proposed that the positive modulation of the hepatic antioxidant systems by ASA may play a role in the chemoprevention of mammary tumourigenesis induced by DMBA in the female rat.
Dexamethasone is a potent synthetic corticosteroid widely employed as a therapeutic agent in catt... more Dexamethasone is a potent synthetic corticosteroid widely employed as a therapeutic agent in cattle. Besides this legal use, corticosteroids are also administered at low dosages as growth-promoters either alone or in combination with other steroids or with beta-agonists. For this reason, appropriate control plans are established to survey corticosteroid misuse, using liver or urine as biological matrices. Since few data are available about the kinetics of dexamethasone excretion in meat cattle, an experimental study was designed to assess the drug residue levels in urines following either a therapeutic (60 microg of dexamethasone sodium phosphate/kg b.w., for three consecutive days) or a growth-promoting schedule (0.7 or 1.4 mg of dexamethasone sodium phosphate per capita/day for 60 days). The urinary elimination of dexamethasone, which was predominantly excreted in the unmodified form, was determined by high-performance liquid chromatography/tandem mass spectrometry at different time intervals, i.e. during the treatments and after appropriate withdrawal times. Our findings confirm the high and rapid rate of dexamethasone urinary excretion irrespective of the nature of the treatment, and provide useful reference values that can be conveniently employed for forensic purposes.
Dietary lipid supplements affect mammary lipid metabolism partly through changes in lipogenic gen... more Dietary lipid supplements affect mammary lipid metabolism partly through changes in lipogenic gene expression. Quantitative PCR (qPCR) is a sensitive, reliable, and accurate technique for gene expression analysis. However, variation introduced in qPCR data by analytical or technical errors needs to be accounted for via normalization using appropriate internal control genes (ICG). Objectives were to mine individual bovine mammary microarray data on >13,000 genes across 66 cows from 2 independent studies to identify the most suitable ICG for qPCR normalization. In addition to unsupplemented control diets, cows were fed saturated or unsaturated lipids for 21 d or were infused with supplements (butterfat, conjugated linoleic acid mixture, long-chain fatty acids) into the abomasum to modify milk fat synthesis and fatty acid profiles. We identified 49 genes that did not vary in expression across the 66 samples. Subsequent gene network analysis revealed that 22 of those genes were not c...
Dioxins and polychlorinated biphenyls (PCBs) are widely spread and long persistent contaminants. ... more Dioxins and polychlorinated biphenyls (PCBs) are widely spread and long persistent contaminants. The aim of this study was to evaluate physiological changes associated with the decontamination of animals previously exposed to environmental pollutants. Eight Limousine heifers were removed from a polluted area and fed a standard ration for six months. The extent of contamination was defined by measuring total toxic equivalents (TEQ) values of dioxin like-PCBs (DL-PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs), and NDL-PCBs amount in pericaudal fat two weeks after the removal from the contaminated area (day 0) and then bimonthly for six months during the decontamination (days 59, 125, and 188). The concentrations of both DL-PCBs + PCDD/Fs and NDL-PCBs at the start of decontamination (day 0) were higher than those legally admitted, and they were strongly decreased at the end of the experimental period. Specific indices of blood redox homeostasis and inflammation were also measured at each time. Serum concentrations of Retinol, Tocopherol and Ascorbate, the total antioxidant capacity (TAC) and the activities of superoxide dismutase and glutathione peroxidase were lower at day 0 than after 59, 125 or 188 days of decontamination. Protein-bound carbonyls (PC), nitro-tyrosine (N-Tyr), and lipid hydroperoxides concentrations were higher at day 0 than during decontamination. In addition, TAC, PC and N-Tyr levels correlated with both DL-PCB and NDL-PCB concentrations only at day 0. Serum concentrations of TNF-alpha and Haptoglobin were higher in samples collected at day 0 than in those obtained during decontamination. As Haptoglobin and TNF-alpha levels correlated with both DL-PCB and NDL-PCB concentrations at day 0 and at day 59 (when these concentrations are still over legal limit), they might represent easily measurable parameters for assessing acute exposure to pollutants. Further both N-Tyr and TNF-alpha concentrations could be used as bio-monitoring markers of the decontamination procedure.
Environmental pollutants, such as dioxin-like (DL) PCBs, benzo(a) pyrene (B[a]P), and flavonoids ... more Environmental pollutants, such as dioxin-like (DL) PCBs, benzo(a) pyrene (B[a]P), and flavonoids are aryl hydrocarbon receptor (AHR) ligands and may be excreted in dairy milk. The expression of AHR-target genes, particularly those involved in xenobiotic biotransformation, and their modulation by two DL-PCBs, B[a]P, and β-naphthoflavone was investigated in a bovine mammary epithelial cell line (BME-UV). As assessed by quantitative PCR, BME-UV cells expressed a functional AHR signaling pathway. All the AHR ligands induced a concentration-related increase in the transcription of cytochrome P450 1A1 and 1B1, known to be implicated in the bioactivation of several xenobiotics. Conversely, genes encoding for antioxidant and detoxifying enzymes, like quinone oxidoreductase or glutathione S-transferase A2, were not affected or even depressed. This study demonstrates the occurrence and the modulation by different AHR-ligands of genes involved in xenobiotic metabolism in BME-UV cells, with the...
Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment, 2015
A chemometric class modelling strategy (unequal dispersed classes - UNEQ) was applied for the fir... more A chemometric class modelling strategy (unequal dispersed classes - UNEQ) was applied for the first time as a possible screening method to monitor the abuse of growth promoters in veal calves. Five serum biomarkers, known to reflect the exposure to classes of compounds illegally used as growth promoters, were determined from 50 untreated animals in order to design a model of controls, representing veal calves reared under good, safe and highly standardised breeding conditions. The class modelling was applied to 421 commercially bred veal calves to separate them into 'compliant' and 'non-compliant' with respect to the modelled controls. Part of the non-compliant animals underwent further histological and chemical examinations to confirm the presence of either alterations in target tissues or traces of illegal substances commonly administered for growth-promoting purposes. Overall, the congruence between the histological or chemical methods and the UNEQ non-compliant o...
7,12-Dimethylbenzanthracene (DMBA) is an abundant environmental contaminant, which undergoes bioa... more 7,12-Dimethylbenzanthracene (DMBA) is an abundant environmental contaminant, which undergoes bioactivation, primarily by the CYP1 family, both in liver and extra-hepatic tissues. Dietary acetylsalicylic acid (ASA) has been recently reported to inhibit DMBA-mediated mammary tumour formation in rats. Chemopreventive substances may reduce the risk of developing cancer by decreasing metabolic enzymes responsible for generating reactive species (phase I enzymes) and/or increasing phase II enzymes that can deactivate radicals and electrophiles. To test these hypotheses, Sprague-Dawley female rats were orally administered ASA as lysine acetylsalicylate (50 mg per capita/day for 21 days in water), DMBA (10 mg per capita in olive oil on day 7, 14, and 21), ASA and DMBA in combination, and vehicles only, respectively. Six rats for each group were sacrificed on day 8, 15, and 22. The DMBA-mediated increase in hepatic CYP1A expression and related activities was not significantly affected by ASA, which, conversely, enhanced in a time-dependent manner the liver reduced glutathione content (up to 52%) and the activity of NAD(P)H-quinone oxidoreductase (up to 34%) in DMBA-treated rats. It is proposed that the positive modulation of the hepatic antioxidant systems by ASA may play a role in the chemoprevention of mammary tumourigenesis induced by DMBA in the female rat.
Dexamethasone is a potent synthetic corticosteroid widely employed as a therapeutic agent in catt... more Dexamethasone is a potent synthetic corticosteroid widely employed as a therapeutic agent in cattle. Besides this legal use, corticosteroids are also administered at low dosages as growth-promoters either alone or in combination with other steroids or with beta-agonists. For this reason, appropriate control plans are established to survey corticosteroid misuse, using liver or urine as biological matrices. Since few data are available about the kinetics of dexamethasone excretion in meat cattle, an experimental study was designed to assess the drug residue levels in urines following either a therapeutic (60 microg of dexamethasone sodium phosphate/kg b.w., for three consecutive days) or a growth-promoting schedule (0.7 or 1.4 mg of dexamethasone sodium phosphate per capita/day for 60 days). The urinary elimination of dexamethasone, which was predominantly excreted in the unmodified form, was determined by high-performance liquid chromatography/tandem mass spectrometry at different time intervals, i.e. during the treatments and after appropriate withdrawal times. Our findings confirm the high and rapid rate of dexamethasone urinary excretion irrespective of the nature of the treatment, and provide useful reference values that can be conveniently employed for forensic purposes.
Dietary lipid supplements affect mammary lipid metabolism partly through changes in lipogenic gen... more Dietary lipid supplements affect mammary lipid metabolism partly through changes in lipogenic gene expression. Quantitative PCR (qPCR) is a sensitive, reliable, and accurate technique for gene expression analysis. However, variation introduced in qPCR data by analytical or technical errors needs to be accounted for via normalization using appropriate internal control genes (ICG). Objectives were to mine individual bovine mammary microarray data on >13,000 genes across 66 cows from 2 independent studies to identify the most suitable ICG for qPCR normalization. In addition to unsupplemented control diets, cows were fed saturated or unsaturated lipids for 21 d or were infused with supplements (butterfat, conjugated linoleic acid mixture, long-chain fatty acids) into the abomasum to modify milk fat synthesis and fatty acid profiles. We identified 49 genes that did not vary in expression across the 66 samples. Subsequent gene network analysis revealed that 22 of those genes were not c...
Dioxins and polychlorinated biphenyls (PCBs) are widely spread and long persistent contaminants. ... more Dioxins and polychlorinated biphenyls (PCBs) are widely spread and long persistent contaminants. The aim of this study was to evaluate physiological changes associated with the decontamination of animals previously exposed to environmental pollutants. Eight Limousine heifers were removed from a polluted area and fed a standard ration for six months. The extent of contamination was defined by measuring total toxic equivalents (TEQ) values of dioxin like-PCBs (DL-PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs), and NDL-PCBs amount in pericaudal fat two weeks after the removal from the contaminated area (day 0) and then bimonthly for six months during the decontamination (days 59, 125, and 188). The concentrations of both DL-PCBs + PCDD/Fs and NDL-PCBs at the start of decontamination (day 0) were higher than those legally admitted, and they were strongly decreased at the end of the experimental period. Specific indices of blood redox homeostasis and inflammation were also measured at each time. Serum concentrations of Retinol, Tocopherol and Ascorbate, the total antioxidant capacity (TAC) and the activities of superoxide dismutase and glutathione peroxidase were lower at day 0 than after 59, 125 or 188 days of decontamination. Protein-bound carbonyls (PC), nitro-tyrosine (N-Tyr), and lipid hydroperoxides concentrations were higher at day 0 than during decontamination. In addition, TAC, PC and N-Tyr levels correlated with both DL-PCB and NDL-PCB concentrations only at day 0. Serum concentrations of TNF-alpha and Haptoglobin were higher in samples collected at day 0 than in those obtained during decontamination. As Haptoglobin and TNF-alpha levels correlated with both DL-PCB and NDL-PCB concentrations at day 0 and at day 59 (when these concentrations are still over legal limit), they might represent easily measurable parameters for assessing acute exposure to pollutants. Further both N-Tyr and TNF-alpha concentrations could be used as bio-monitoring markers of the decontamination procedure.
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