This study examined the ability of several human pharmaceuticals to modulate hepatic piscine CYP-mediated monooxygenase activities. Effects of six pharmaceuticals: diclofenac, sulfamethoxazole, tramadol, carbamazepine, venlafaxine and nefazodone, were investigated in vitro in rainbow trout hepatic microsomes. The reactions of 7-ethoxyresorufin-O-deethylase (EROD) and benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylase (BFCOD), were used as markers for hepatic CYP1A and CYP3A-like activities, respectively. Our results showed that EROD and BFCOD activities were both affected by nefazodone. Nefazodone inhibited EROD in a dose dependent manner and was found to be a potent non-competitive inhibitor of EROD with a Ki value of 6.6 μM. BFCOD activity was inhibited non-competitively in the presence of nefazadone with Ki value of 30.7 μM. BFCOD activity was slightly reduced only by the highest concentration of carbamazepine. Diclofenac, sulfamethoxazole, tramadol, and venlafaxine did not affect the activity of either EROD or BFCOD. We further exposed microsomal fraction to mixtures of six pharmaceuticals to investigate potential inhibition. The results showed that EROD and BFCOD activity was inhibited on 94% and 80%, respectively at higher tested concentration. To our knowledge, this is the first report to demonstrate an inhibitory effect of nefazodone on hepatic CYP1A and CYP3A-like proteins in rainbow trout.

• Klíčová slova
• Antidepressant, Cytochrome P450, Fish microsomes, Inhibition, Mixture of compounds,
• MeSH
• chemické látky znečišťující vodu MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• cytochrom P-450 CYP3A metabolismus   MeSH
• jaterní mikrozomy enzymologie   metabolismus   MeSH
• játra metabolismus   MeSH
• léčivé přípravky metabolismus   MeSH
• lidé MeSH
• Oncorhynchus mykiss metabolismus   MeSH
• piperaziny MeSH
• triazoly farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• cytochrom P-450 CYP1A1 MeSH
• cytochrom P-450 CYP3A MeSH
• léčivé přípravky MeSH
• nefazodone MeSH Prohlížeč
• piperaziny MeSH
• triazoly MeSH

Efficiency of advanced wastewater treatment technologies to reduce micropollutants which mediate dioxin-like toxicity was investigated. Technologies compared included ozonation, powdered activated carbon and granular activated carbon. In addition to chemical analyses in samples of effluents, surface waters, sediments, and fish, (1) dioxin-like potentials were measured in paired samples of effluents, surface waters, and sediments by use of an in vitro biotest (reporter gene assay) and (2) dioxin-like effects were investigated in exposed fish by use of in vivo activity of the mixed-function, monooxygenase enzyme, ethoxyresorufin O-deethylase (EROD) in liver. All advanced technologies studied, based on degradation or adsorption, significantly reduced dioxin-like potentials in samples and resulted in lesser EROD activity in livers of fish. Results of in vitro and in vivo biological responses were not clearly related to quantification of targeted analytes by use of instrumental analyses.

• Klíčová slova
• Activated carbon, EROD, Ozonation, Reporter gene assay, Sewage,
• MeSH
• adsorpce MeSH
• biotest MeSH
• chemické látky znečišťující vodu analýza   MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• dioxiny chemie   toxicita   MeSH
• dřevěné a živočišné uhlí MeSH
• játra enzymologie   MeSH
• nemoci ryb chemicky indukované   MeSH
• odpadní voda analýza   MeSH
• regulace genové exprese enzymů MeSH
• ryby MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• cytochrom P-450 CYP1A1 MeSH
• dioxiny MeSH
• dřevěné a živočišné uhlí MeSH
• odpadní voda MeSH

Naturally- and anthropogenically-produced cresols could pose serious risks to fish health. In this study, three piscine CYP isoforms were investigated for their abilities to interact with p-cresol. Therefore, the activity of 7-ethoxyresorufin-O-deethylase (EROD), 7-benzyloxy-4-trifluoromethylcoumarin O-debenzylase (BFCOD), and p-nitrophenol hydroxylase (PNPH) were evaluated in the hepatic microsomes of juvenile rainbow trout. Results showed that EROD activity was inhibited in a competitive manner, BFCOD activity was inhibited in presence the highest tested p-cresol concentration and PNPH activity was not affected. These results indicate that p-cresol might affect the ability of fish to metabolize numerous aromatic hydrocarbons and dioxin compounds, which are present in the aquatic environment.

• Klíčová slova
• CYP1A1, CYP2E1-like, CYP3A-like, Fish, p-Cresol,
• MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• cytochrom P-450 CYP1A1 antagonisté a inhibitory   metabolismus   MeSH
• kresoly toxicita   MeSH
• Oncorhynchus mykiss metabolismus   MeSH
• rybí proteiny metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 4-cresol MeSH Prohlížeč
• chemické látky znečišťující vodu MeSH
• cytochrom P-450 CYP1A1 MeSH
• kresoly MeSH
• rybí proteiny MeSH

The RTgill-W1 (gill), RTG-2 (gonad), and RTL-W1 (liver) cell lines derived from a freshwater fish rainbow trout (Oncorhynchus mykiss), were used to assess the toxicity of polyethylene terephthalate (PET) and two forms of polyvinyl chloride (PVC). Two size fractions (25-μm and 90-μm particles) were tested for all materials. The highest tested concentration was 1 mg/ml, corresponding to from 70 000 ± 9000 to 620 000 ± 57 000 particles/ml for 25-μm particles and from 2300 ± 100 to 11 000 ± 1000 particles/ml for 90-μm particles (depending on the material). Toxicity differences between commercial PVC dry blend powder and secondary microplastics created from a processed PVC were newly described. After a 24-h exposure, the cells were analyzed for changes in viability, 7-ethoxyresorufin-O-deethylase (EROD) activity, and reactive oxygen species (ROS) generation. In addition to the microplastic suspensions, leachates and particles remaining after leaching resuspended in fresh exposure medium were tested. The particles were subjected to leaching for 1, 8, and 15 days. The PVC dry blend (25 μm and 90 μm) and processed PVC (25 μm) increased ROS generation, to which leached chemicals appeared to be the major contributor. PVC dry blend caused substantially higher ROS induction than processed PVC, showing that the former is not suitable for toxicity testing, as it can produce different results from those of secondary PVC. The 90-μm PVC dry blend increased ROS generation only after prolonged leaching. PET did not induce any changes in ROS generation, and none of the tested polymers had any effect on viability or EROD activity. The importance of choosing realistic extraction procedures for microplastic toxicity experiments was emphasized. Conducting long-term experiments is crucial to detect possible environmentally relevant effects. In conclusion, the tested materials showed no acute toxicity to the cell lines.

• Klíčová slova
• Microplastics, Oncorhynchus mykiss, PET, PVC, Reactive oxygen species, Toxicity,
• MeSH
• buněčné linie MeSH
• chemické látky znečišťující vodu * analýza   MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• mikroplasty toxicita   MeSH
• Oncorhynchus mykiss * metabolismus   MeSH
• plastické hmoty toxicita   metabolismus   MeSH
• polyethylentereftaláty toxicita   metabolismus   MeSH
• polyvinylchlorid toxicita   metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• cytochrom P-450 CYP1A1 MeSH
• mikroplasty MeSH
• plastické hmoty MeSH
• polyethylentereftaláty MeSH
• polyvinylchlorid MeSH
• reaktivní formy kyslíku MeSH

Effects of aquatic pollutants on fish are of increasing concern. Pharmaceutical-based contaminants are prioritized for further study in environmental risk assessment using several approaches. Dexamethasone (DEX) was one such contaminant recognised for its effect on fish health status. Thus, we carried out an in vivo experiment to identify potential effects of DEX on rainbow trout. Fish were exposed to 3, 30, 300 and 3000ngL(-1) DEX in a semi-static system over a period of 42d. The concentrations of DEX that fish were exposed to was confirmed by LC-LC-MS/MS. Using hepatic microsomes, we determined cytochrome P450 content, activities of ethoxyresorufin O-deethylase (EROD), p-nitrophenol hydroxylase (PNPH), 7-benzyloxy-4-trifluoromethylcoumarin O-debenzylase (BFCOD) and benzyloxyquinoline O-debenzylase (BQOD), as well as protein expression. Our results showed that fish do not change the catalytic activity of CYP450-mediated reactions after high DEX concentration exposure. These results disagree with mammalian studies, where DEX is a well-known inducer of CYP450. We showed a significant effect of DEX exposure on CYP450-mediated reactions (EROD, BCFOD, BQOD and PNPH) when expressed as amount of product formed per min per nmol total CYP450 at 3, 30 and 300ngL(-1) after 21d exposure. Moreover, BFCOD and BQ activities showed matching trends in all groups. Western blot analysis showed induction of CYP3A-like protein in the presence of the lowest environmentally relevant concentration of DEX. Based on these findings, continued investigation of the effect of DEX on fish using a battery of complementary biomarkers of exposure and effect is highly relevant.

• Klíčová slova
• CYP1A, CYP2E1-like protein, CYP3A, Dexamethasone, Rainbow trout,
• MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• dexamethason toxicita   MeSH
• jaterní mikrozomy účinky léků   enzymologie   MeSH
• Oncorhynchus mykiss metabolismus   MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• cytochrom P-450 CYP1A1 MeSH
• dexamethason MeSH
• systém (enzymů) cytochromů P-450 MeSH

Piscine cytochrome P450 (CYP) enzymes play an important role in the metabolism of xenobiotics. Xenobiotics often act as inducers of CYP1A1 and CYP3A expression and activity in fish. We compared constitutive mRNA expression of CYP1A1, CYP3A27, and CYP3A45 and catalytic activity of CYP1A (7-ethoxyresorufin-O-deethylation, EROD) and CYP3A-like (benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylation, BFCOD) enzymes in the following six rainbow trout tissues: liver, gill, heart, brain, intestine, and gonad. mRNA expression and activity were present in all investigated tissues. The CYP1A1 mRNA expression was higher in the liver, gill, heart, and brain compared to gonad and intestine. The intestine was the main site of CYP3A27 and CYP3A45 expression. The highest EROD and BFCOD activity was observed in liver tissue followed in descending order by heart, brain, gill, intestine, and gonad. Such differences might be related to the role of CYP physiological functions in the specific tissue. Rainbow trout exposure to 50 mg/kg of β-naphthoflavone for 48 h resulted in a 7.5- and 5.9-fold increase in liver EROD and BFCOD activity, respectively. In vitro EROD activity inhibition with ellipticine showed tissue-specific inhibition, while ketoconazole decreased BFCOD activity by 50-98 % in all tissues. Further studies are needed to identify all CYP isoforms that are responsible for these activities and modes of regulation.

• Klíčová slova
• CYP inhibitors, CYP1A1, CYP3A, Catalytic activity, P450,
• MeSH
• cytochrom P-450 CYP1A1 genetika   metabolismus   MeSH
• cytochrom P-450 CYP3A genetika   metabolismus   MeSH
• játra enzymologie   MeSH
• messenger RNA genetika   metabolismus   MeSH
• mozek enzymologie   MeSH
• myokard enzymologie   MeSH
• Oncorhynchus mykiss metabolismus   MeSH
• pohlavní dimorfismus MeSH
• regulace genové exprese enzymů fyziologie   MeSH
• střeva enzymologie   MeSH
• žábry enzymologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytochrom P-450 CYP1A1 MeSH
• cytochrom P-450 CYP3A MeSH
• messenger RNA MeSH

The hepatic cytochrome p450 enzymes 1 A, 2A19 and 2E1 is very important for the elimination of skatole from the body of pigs. Impaired skatole metabolism, results in skatole accumulation, which give rise to off flavor of the meat. Several metabolites of skatole has been identified, however the role of these metabolites in the inhibition of the skatole metabolizing enzymes are not documented. Using microsomes from pigs and fish, we determined the ability of several skatole metabolites to inhibit CYP1 A, CYP2A19 and CYP2E1 dependent activity. Our results show that 2-aminoacetophenone is an inhibitor of porcine CYP2A19 and CYP2E1 activity, but not the piscine orthologues. In conclusion, there is species specific differences in the inhibition of CYP1 A and CYP2A19 dependent metabolism of probe substrates. This is relevant to the evaluation of different model systems and to the reduction of off flavor of meat.

• Klíčová slova
• Cytochrome P450, Pig, Rainbow trout, Skatole metabolism,
• MeSH
• acetofenony toxicita   MeSH
• červené maso analýza   MeSH
• cytochrom P-450 CYP1A1 antagonisté a inhibitory   metabolismus   MeSH
• cytochrom P-450 CYP2E1 metabolismus   MeSH
• druhová specificita MeSH
• inhibitory cytochromu P450 CYP2E1 toxicita   MeSH
• jaterní mikrozomy účinky léků   metabolismus   MeSH
• játra účinky léků   metabolismus   MeSH
• kumariny toxicita   MeSH
• nitrofenoly toxicita   MeSH
• oxaziny toxicita   MeSH
• potrava z moře (živočišná) analýza   MeSH
• prasata MeSH
• ryby MeSH
• skatol toxicita   MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 2-aminoacetophenone MeSH Prohlížeč
• 4-nitrophenol MeSH Prohlížeč
• acetofenony MeSH
• coumarin MeSH Prohlížeč
• cytochrom P-450 CYP1A1 MeSH
• cytochrom P-450 CYP2E1 MeSH
• ethoxyresorufin MeSH Prohlížeč
• inhibitory cytochromu P450 CYP2E1 MeSH
• kumariny MeSH
• nitrofenoly MeSH
• oxaziny MeSH
• skatol MeSH
• systém (enzymů) cytochromů P-450 MeSH

Modulations of 11 prospective biochemical markers of impacts of aquatic pollutants in liver tissue of chub (Leuciscus cephalus), caught at several sampling sites of a river with various pollution types and rates, were matched against analytical data of concentrations of organochlorine compounds, polycyclic aromatic hydrocarbons (PAHs), and heavy metals. Multivariate principal component analysis (PCA) of the field data showed general patterns of biochemical responses to different types of pollutants and relationships among the biomarkers. Cytochrome P4501A-dependent 7-ethoxyresorufin O-deethylase (EROD) activity, inducible by 2,3,7,8-tetrachlorodibenzo-p-dioxin and structurally related planar compounds, was strongly enhanced in the more contaminated areas. Compared with polychlorinated aromatic hydrocarbons, PAHs did not contribute so significantly to EROD induction. Testosterone 6 beta- and 16 alpha-hydroxylase activities, as an expression of the cytochrome P4503A27, were slightly increased at several sites but were significantly decreased in samples from some heavily polluted areas. Recently, these activities have been suggested as potential biomarkers of exposure to contaminants that do not induce cytochrome P4501A. In this study, their inhibition or induction was not associated with a specific class of monitored contaminants, and selectivities of these modulations are still to be investigated. Similar modulations of the prospective biochemical indicators of oxidative stress, including microsomal glutathione S-transferase activity, cytosolic glutathione S-transferase with ethacrynic acid, and glutathione reductase, were demonstrated by PCA. The pattern of the modulations of the microsomal nicotinamide adenine dinucleotide phosphate (NADPH)-dependent lipid peroxidation in vitro differed from the responses of the rest of oxidative stress parameters at some sampling sites. Further biochemical markers of oxidative stress under study, including in vivo lipid peroxidation, in vitro production of reactive oxygen species, and the concentration of metallothioneins did not correlate well with the concentrations of the contaminants. Principal component analysis demonstrated that the EROD activity, glutathione-dependent enzymes, and Fe(II)-enhanced lipid peroxidation formed a suitable battery of biomarkers of exposure.

• MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• chlorované uhlovodíky toxicita   MeSH
• Cyprinidae metabolismus   MeSH
• cytochrom P-450 CYP1A1 biosyntéza   metabolismus   MeSH
• enzymová indukce MeSH
• játra účinky léků   embryologie   MeSH
• multivariační analýza MeSH
• peroxidace lipidů účinky léků   MeSH
• polycyklické sloučeniny toxicita   MeSH
• steroid-16-alfa-hydroxylasa MeSH
• těžké kovy toxicita   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• chlorované uhlovodíky MeSH
• cytochrom P-450 CYP1A1 MeSH
• polycyklické sloučeniny MeSH
• steroid-16-alfa-hydroxylasa MeSH
• těžké kovy MeSH

The effects of inclusion of sesamin / episesamin in Baltic Atlantic salmon (Salmo salar L.) diets based on vegetable oils were studied. The study was designed as a dose response study with two control diets, one diet based on fish oil (FO) and one diet based on a mixture of linseed and sunflower oil (6:4 by vol.) (MO). As experimental diets three different levels of inclusion of sesamin / episesamin (hereafter named sesamin) to the MO based diet and one diet based on sesame oil and linseed oil (SesO) (1:1 by vol.) were used. The dietary oils were mirrored in the fatty acid profile of the white muscle. Sesamin significantly decreased the levels of 18:3n-3 in the white muscle phospholipid (PL) fraction of all groups fed sesamin, no significant differences were found in the triacylglycerol fraction (TAG). Slightly increased levels of docosahexaenoic acid (22:6n-3, DHA) in PL and TAG were found in some of the sesamin fed groups. Sesamin significantly affected the expression of peroxisome proliferator-activated receptor alpha, scavenger receptor type B and hormone sensitive lipase, in agreement with previous studies on rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar L.) hepatocytes published by our group. No significant effects on toxicological response measured as ethoxyresorufin O-deethylase activity was found. The total cytochrome P450 enzymes were significantly higher in MO 0.29 and SesO group. The amount of alpha- and gamma-tocopherols in liver and the amount of gamma-tocopherol in white muscle were significantly lower in fish fed the FO diet compared to the MO diet, but no difference after inclusion of sesamin was found in this study. Increased inclusion of sesamin increased the levels of sesamin and episesamin in the liver, but did not affect the amounts in white muscle.

• MeSH
• cytochrom P-450 CYP1A1 genetika   metabolismus   MeSH
• dioxoly aplikace a dávkování   metabolismus   MeSH
• játra metabolismus   MeSH
• lignany aplikace a dávkování   metabolismus   MeSH
• mastné kyseliny metabolismus   MeSH
• metabolismus lipidů genetika   MeSH
• Salmo salar genetika   metabolismus   MeSH
• tokoferoly metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytochrom P-450 CYP1A1 MeSH
• dioxoly MeSH
• lignany MeSH
• mastné kyseliny MeSH
• sesamin MeSH Prohlížeč
• tokoferoly MeSH

Targeting the aryl hydrocarbon receptor (AhR) is an emerging therapeutic strategy for multiple diseases (e.g., inflammatory bowel disease). Thermosporothrix hazakensis microbial metabolite 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE) is a putative AhR endogenous ligand. To improve the chemical stability, we synthesized a series of ITE chemical mimics. Using a series of in vitro assays, we identified 2-(1H-indole-3-carbonyl)-N-methyl thiazole-4-carboxamide (ITE-CONHCH3) as a highly potent (EC50 = 1.6 nM) AhR agonist with high affinity (Ki = 88 nM). ITE-CONHCH3 triggered AhR nuclear translocation and dimerization of AhR-ARNT, enhanced AhR binding in the CYP1A1 promoter, and induced AhR-regulated genes in an AhR-dependent manner. The metabolic stability of ITE-CONHCH3 in a cell culture was 10 times higher than that of ITE. Finally, we observed protective effects of ITE-CONHCH3 in mice with DSS-induced colitis. Overall, we demonstrate and validate a concept of microbial metabolite mimicry in the therapeutic targeting of AhR.

• MeSH
• cytochrom P-450 CYP1A1 MeSH
• indoly farmakologie   terapeutické užití   MeSH
• kolitida * chemicky indukované   farmakoterapie   MeSH
• myši MeSH
• receptory aromatických uhlovodíků * agonisté   genetika   metabolismus   MeSH
• thiazoly farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytochrom P-450 CYP1A1 MeSH
• indoly MeSH
• receptory aromatických uhlovodíků * MeSH
• thiazoly MeSH