title: Assessing the potential of pharmaceuticals and their transformation products to cause mutagenic effects: Implications for gene expression profiling.
abstract: The selection and prioritization of pharmaceuticals and their transformation products for evaluating effects on the environment and human health is a challenging task. One common approach is based on compounds (e.g., mixture composition, concentrations), and another on biology (e.g., relevant endpoint, biological organizational level). Both of these approaches often resemble a Lernaean Hydra-they can create more questions than answers. The present study embraces this complexity, providing an integrated approach toward assessing the potential effects of transformation products of pharmaceuticals by means of mutagenicity, estrogenicity, and differences in the gene expression profiles. Mutagenicity using the tk kinase assay was applied to assess a list of 11 priority pharmaceuticals, namely, atenolol, azithromycin, carbamazepine, diclofenac, ibuprofen, erythromycin, metoprolol, ofloxacin, propranolol, sulfamethoxazole, and trimethoprim. The most mutagenic compounds were found to be β-blockers. In parallel, the photolabile pharmaceuticals were assessed for their mixture effects on mutagenicity (tk assay), estrogenicity (T47D- KBluc assay), and gene expression (microarrays). Interestingly, the mixtures were mutagenic at the µg/L level, indicating a synergistic effect. None of the photolysed mixtures were statistically significantly estrogenic. Gene expression profiling revealed effects related mainly to certain pathways, those of the p53 gene, mitogen-activated protein kinase, alanine, aspartate, and glutamate metabolism, and translation-related (spliceosome). Fourteen phototransformation products are proposed based on the m/z values found through ultra-performance liquid chromatography-tandem mass spectrometry analysis. The transformation routes of the photolysed mixtures indicate a strong similarity with those obtained for each pharmaceutical separately. This finding reinforces the view that transformation products are to be expected in naturally occurring mixtures. Environ Toxicol Chem 2016;35:2753-2764. © 2016 SETAC.
authors.affiliation: Nireas-International Water Research Center, University of Cyprus, Nicosia, Cyprus.
authors.forename: Marlen I
authors.initials: MI
authors.lastname: Vasquez
authors.affiliation: Nireas-International Water Research Center, University of Cyprus, Nicosia, Cyprus.|Department of Civil and Environmental Engineering, University of Cyprus, Nicosia, Cyprus.
authors.forename: Maria
authors.initials: M
authors.lastname: Tarapoulouzi
authors.affiliation: Nireas-International Water Research Center, University of Cyprus, Nicosia, Cyprus.|The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus.
authors.forename: Nancy
authors.initials: N
authors.lastname: Lambrianides
authors.affiliation: Nireas-International Water Research Center, University of Cyprus, Nicosia, Cyprus.
authors.forename: Evroula
authors.initials: E
authors.lastname: Hapeshi
authors.affiliation: Department of Life & Health Sciences, University of Nicosia, Nicosia, Cyprus.
authors.forename: Kyriakos
authors.initials: K
authors.lastname: Felekkis
authors.affiliation: Medical Research Center, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Saile
authors.affiliation: Medical Research Center, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Sticht
authors.affiliation: Medical Research Center, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.
authors.forename: Norbert
authors.initials: N
authors.lastname: Gretz
authors.affiliation: Nireas-International Water Research Center, University of Cyprus, Nicosia, Cyprus. dfatta@ucy.ac.cy.|Department of Civil and Environmental Engineering, University of Cyprus, Nicosia, Cyprus. dfatta@ucy.ac.cy.
authors.forename: Despo
authors.initials: D
authors.lastname: Fatta-Kassinos
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000032:analysis / Q000528:radiation effects / Q000633:toxicity*; D000818:Animals; D045744:Cell Line, Tumor; D045424:Complex Mixtures / Q000032:analysis / Q000528:radiation effects / Q000633:toxicity*; D020869:Gene Expression Profiling; D006801:Humans; D051379:Mice; D046228:Microarray Analysis; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D009153:Mutagens / Q000032:analysis / Q000528:radiation effects / Q000633:toxicity*; D015242:Ofloxacin; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000528:radiation effects; D010782:Photolysis; D059467:Transcriptome / Q000187:drug effects*; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000528:radiation effects / Q000633:toxicity*
chemical_list: D000319:Adrenergic beta-Antagonists; D045424:Complex Mixtures; D009153:Mutagens; D004364:Pharmaceutical Preparations; D014874:Water Pollutants, Chemical; D015242:Ofloxacin; D020928:Mitogen-Activated Protein Kinases
keywords: Gene expression; Mammalian toxicology; Mixture toxicity; Mutagenicity; Pharmaceutical; Transformation product