- Email: [email protected]
Acrylamide induced chronic nephrotoxicity in rats
S234
Abstracts / Toxicology Letters 221S (2013) S59–S256
readily be exposed to chemical and pharmaceutical compounds and are susceptible to genetic manipulation. One potential limitation, however, is the functional divergence of their bioactivation and detoxification systems compared to those of preclinical mammalian models used in drug development. To circumvent this limitation we are generating “humanized” transgenic zebrafish that mimic the human response to xenobiotics. Reference compounds with known or no in vivo liver toxicity were tested on larval and adult zebrafish as well as isolated liver microsomes and the effects compared to available in vitro (hepatocyte) and in vivo (clinical/preclinical) data. Uptake and metabolism of compounds was analysed by LC/MS. Testing with human CYP specific substrates indicated that the wild-type zebrafish CYP3A substrate specificity differs from that of its human counterpart CYP3A4. A BAC containing human CYP genes was modified for Tol2 mediated transgenesis and used to generate a stable humanized CYP3A4 transgenic line. Enzyme activity studies with the human CYP3A4 substrate, midazolam, showed a significant increase in 1hdyroxy-midazolam metabolite generation in the humanized fish compared to wild-type. To identify novel biomarkers that respond to a broad spectrum of xenobiotics a genome-wide transcriptome analysis of liver was performed following exposure to selected compounds. Treated samples showed clear clusters of differential expression relative to controls and a number of up-regulated genes were selected for further analysis. Transgenic reporters of their expression are being generated to facilitate rapid screening of compounds. http://dx.doi.org/10.1016/j.toxlet.2013.05.570
P23-03 Acrylamide induced chronic nephrotoxicity in rats Ahmad Raoufi, Keivan Jamshidi ∗ IAU Garmsar Branch, Garmsar, Iran Acrylamide (ACR) has been shown to cause neurotoxic effects in humans and neurotoxic, genotoxic, reproductive, and carcinogenic effects in laboratory animals. To investigate the nephrotoxic effects of ACR, 40 adult male rats (Wistar, approximately 250 g) randomly assigned in 4 groups; including 3 treatment groups and 1 control group named as A, B, C, and D respectively. Rats in treatment groups were exposed to 0.1, 1, and 10 mg/kg ACR per day × 90 days p.o (gavage), respectively. The remaining 10 rats in control group received daily p.o (gavage) of 0.9% saline (3 ml/kg). On day 91, two rats, were randomly selected, perfused, dissected and proper samples were collected from their kidneys. Results of histopathological studies based on H&E technique did show no morphologic changes in kidneys of rats belong to groups A, B and D, while moderate to severe morphologic changes including glomerular hypercellularity, global pattern of proliferative glomerulonephritis, occupation of capsular space, tubular cell swelling and hyaline cast formation, were observed in different stained sections obtained from the kidneys of rats belong to group C. This finding, beside neurotoxic, reproductive and carcinogenic effects, appears to indicate for the first time another important aspect of toxic effect of ACR, i.e., chronic nephrotoxicity. http://dx.doi.org/10.1016/j.toxlet.2013.05.571
P23-04 Chloracne/MADISH-like cystic lesions after low-dose exposure to dioxin: A histological study of 43 cases Nikolina Saxer-Sekulic 1,∗ , Gürkan Kaya 1 , Jean-Hilaire Saurat 2 , Olivier Sorg 2 1
Department of Dermatology, University Hospital of Geneva, Switzerland, 2 Swiss Center for Applied Human Toxicology, University of Geneva, Switzerland A pollutant waste incinerator was exploited in Melun area in south of Paris for 28 years. Dioxin analysis of the fumes released by the incinerator, the soil, the eggs and the blood of several inhabitants indicated that this population had been exposed to dioxin-like compounds over a long period of time. Here we describe the morphological analysis of 43 cases (23 m, 20 f) from this population. Hematoxylin–eosin-stained sections from retroauricular and lesional skin biopsies were analysed. On histological examination, diagnosis of normal skin was made for 6 of these patients. 22 patients showed histologically perifolliculitis, with a nonspecific dermal inflammatory infiltrate in two patients, follicular eczema in one patient and eosinophilic infiltrate in one patient. Inflammatory infiltrate was composed of lymphocytes, sometimes in addition with some neutrophils or eosinophils. Follicular wall was not destructed in any of these cases. On histological examination, 4 of the 43 patients showed cystic lesions compatible with chloracne/MADISH, three of them were also detected clinically. The impact on health on long-term exposure to subtoxic doses of dioxin-like compounds is unknown. Here we show in a population exposed for a long time to low amounts of dioxin from the environment some cutaneous morphological changes compatible with those observed in acute dioxin intoxication. By completing this analysis with immunohistochemical labelling such as CYP1A1, it would be possible to use the morphological analysis of skin biopsies as a biomarker of chronic exposure to low dose of dioxin-like pollutants. http://dx.doi.org/10.1016/j.toxlet.2013.05.572
P23-05 Comparison of the accumulation of ochratoxin A residues in treated pig tissues Jelka Pleadin a,∗ , Nina Perˇsi a , Dragan Kovaˇcevic´ b , Ana Vulic´ a , Dinka Milic´ c a
Croatian Veterinary Institute Zagreb, Laboratory for Analytical Chemistry, Savska cesta 143, 10000 Zagreb, Croatia, b University of Josip Juraj Strossmayer in Osijek, Faculty of Food Technology, Franje Kuhaˇca 20, 31000 Osijek, Croatia, c Dubravica Swine Farm, Ltd., Pavla Sˇ toosa 109, 10293 Dubravica, Croatia Driven by the observation that the mycotoxin ochratoxin A (OTA) often contaminates cereals and feed in this European region, the aim of the study was to compare the accumulation of OTA residues in different pig tissues exposed to sub-chronic treatment. The treated animal group (n = 5) was administered 300 g OTA/kg of feed during a 30-day period, whereas the control group (n = 5) was left untreated. OTA concentrations were quantified using high performance liquid chromatography with fluorescence detection (HPLC-FD) that had shown acceptable validation results when it comes to all investigated tissues, with mean recoveries spanning from 88.2% to 96.1% and acceptable mean inter- and intra-day
Abstracts / Toxicology Letters 221S (2013) S59–S256
readily be exposed to chemical and pharmaceutical compounds and are susceptible to genetic manipulation. One potential limitation, however, is the functional divergence of their bioactivation and detoxification systems compared to those of preclinical mammalian models used in drug development. To circumvent this limitation we are generating “humanized” transgenic zebrafish that mimic the human response to xenobiotics. Reference compounds with known or no in vivo liver toxicity were tested on larval and adult zebrafish as well as isolated liver microsomes and the effects compared to available in vitro (hepatocyte) and in vivo (clinical/preclinical) data. Uptake and metabolism of compounds was analysed by LC/MS. Testing with human CYP specific substrates indicated that the wild-type zebrafish CYP3A substrate specificity differs from that of its human counterpart CYP3A4. A BAC containing human CYP genes was modified for Tol2 mediated transgenesis and used to generate a stable humanized CYP3A4 transgenic line. Enzyme activity studies with the human CYP3A4 substrate, midazolam, showed a significant increase in 1hdyroxy-midazolam metabolite generation in the humanized fish compared to wild-type. To identify novel biomarkers that respond to a broad spectrum of xenobiotics a genome-wide transcriptome analysis of liver was performed following exposure to selected compounds. Treated samples showed clear clusters of differential expression relative to controls and a number of up-regulated genes were selected for further analysis. Transgenic reporters of their expression are being generated to facilitate rapid screening of compounds. http://dx.doi.org/10.1016/j.toxlet.2013.05.570
P23-03 Acrylamide induced chronic nephrotoxicity in rats Ahmad Raoufi, Keivan Jamshidi ∗ IAU Garmsar Branch, Garmsar, Iran Acrylamide (ACR) has been shown to cause neurotoxic effects in humans and neurotoxic, genotoxic, reproductive, and carcinogenic effects in laboratory animals. To investigate the nephrotoxic effects of ACR, 40 adult male rats (Wistar, approximately 250 g) randomly assigned in 4 groups; including 3 treatment groups and 1 control group named as A, B, C, and D respectively. Rats in treatment groups were exposed to 0.1, 1, and 10 mg/kg ACR per day × 90 days p.o (gavage), respectively. The remaining 10 rats in control group received daily p.o (gavage) of 0.9% saline (3 ml/kg). On day 91, two rats, were randomly selected, perfused, dissected and proper samples were collected from their kidneys. Results of histopathological studies based on H&E technique did show no morphologic changes in kidneys of rats belong to groups A, B and D, while moderate to severe morphologic changes including glomerular hypercellularity, global pattern of proliferative glomerulonephritis, occupation of capsular space, tubular cell swelling and hyaline cast formation, were observed in different stained sections obtained from the kidneys of rats belong to group C. This finding, beside neurotoxic, reproductive and carcinogenic effects, appears to indicate for the first time another important aspect of toxic effect of ACR, i.e., chronic nephrotoxicity. http://dx.doi.org/10.1016/j.toxlet.2013.05.571
P23-04 Chloracne/MADISH-like cystic lesions after low-dose exposure to dioxin: A histological study of 43 cases Nikolina Saxer-Sekulic 1,∗ , Gürkan Kaya 1 , Jean-Hilaire Saurat 2 , Olivier Sorg 2 1
Department of Dermatology, University Hospital of Geneva, Switzerland, 2 Swiss Center for Applied Human Toxicology, University of Geneva, Switzerland A pollutant waste incinerator was exploited in Melun area in south of Paris for 28 years. Dioxin analysis of the fumes released by the incinerator, the soil, the eggs and the blood of several inhabitants indicated that this population had been exposed to dioxin-like compounds over a long period of time. Here we describe the morphological analysis of 43 cases (23 m, 20 f) from this population. Hematoxylin–eosin-stained sections from retroauricular and lesional skin biopsies were analysed. On histological examination, diagnosis of normal skin was made for 6 of these patients. 22 patients showed histologically perifolliculitis, with a nonspecific dermal inflammatory infiltrate in two patients, follicular eczema in one patient and eosinophilic infiltrate in one patient. Inflammatory infiltrate was composed of lymphocytes, sometimes in addition with some neutrophils or eosinophils. Follicular wall was not destructed in any of these cases. On histological examination, 4 of the 43 patients showed cystic lesions compatible with chloracne/MADISH, three of them were also detected clinically. The impact on health on long-term exposure to subtoxic doses of dioxin-like compounds is unknown. Here we show in a population exposed for a long time to low amounts of dioxin from the environment some cutaneous morphological changes compatible with those observed in acute dioxin intoxication. By completing this analysis with immunohistochemical labelling such as CYP1A1, it would be possible to use the morphological analysis of skin biopsies as a biomarker of chronic exposure to low dose of dioxin-like pollutants. http://dx.doi.org/10.1016/j.toxlet.2013.05.572
P23-05 Comparison of the accumulation of ochratoxin A residues in treated pig tissues Jelka Pleadin a,∗ , Nina Perˇsi a , Dragan Kovaˇcevic´ b , Ana Vulic´ a , Dinka Milic´ c a
Croatian Veterinary Institute Zagreb, Laboratory for Analytical Chemistry, Savska cesta 143, 10000 Zagreb, Croatia, b University of Josip Juraj Strossmayer in Osijek, Faculty of Food Technology, Franje Kuhaˇca 20, 31000 Osijek, Croatia, c Dubravica Swine Farm, Ltd., Pavla Sˇ toosa 109, 10293 Dubravica, Croatia Driven by the observation that the mycotoxin ochratoxin A (OTA) often contaminates cereals and feed in this European region, the aim of the study was to compare the accumulation of OTA residues in different pig tissues exposed to sub-chronic treatment. The treated animal group (n = 5) was administered 300 g OTA/kg of feed during a 30-day period, whereas the control group (n = 5) was left untreated. OTA concentrations were quantified using high performance liquid chromatography with fluorescence detection (HPLC-FD) that had shown acceptable validation results when it comes to all investigated tissues, with mean recoveries spanning from 88.2% to 96.1% and acceptable mean inter- and intra-day