Peroxisome proliferator-activated receptor alpha (PPARα) is a molecular target for perfluoroalkyl substances (PFASs). Little is known about the cellular uptake of PFASs and how it affects the PPARα activity. We investigated the relationship between PPARα activity and cellular concentration in HepG2 cells of 14 PFASs, including perfluoroalkyl carboxylates (PFCAs), perfluoroalkyl sulfonates and perfluorooctane sulfonamide (FOSA). Cellular concentrations were determined by high-performance liquid chromatography-tandem mass spectrometry and PPARα activity was determined in transiently transfected cells by reporter gene assay. Cellular uptake of the PFASs was low (0.04-4.1%) with absolute cellular concentrations in the range 4-2500 ng mg protein. Cellular concentration of PFCAs increased with perfluorocarbon chain length up to perfluorododecanoate. PPARα activity of PFCAs increased with chain length up to perfluorooctanoate. The maximum induction of PPARα activity was similar for short-chain (perfluorobutanoate and perfluoropentanoate) and long-chain PFCAs (perfluorododecanoate and perfluorotetradecanoate) (approximately twofold). However, PPARα activities were induced at lower cellular concentrations for the short-chain homologs compared to the long-chain homologs. Perfluorohexanoate, perfluoroheptanoate, perfluorooctanoate, perfluorononanoate (PFNA) and perfluorodecanoate induced PPARα activities >2.5-fold compared to controls. The concentration-response relationships were positive for all the tested compounds, except perfluorooctane sulfonate PFOS and FOSA, and were compound-specific, as demonstrated by differences in the estimated slopes. The relationships were steeper for PFCAs with chain lengths up to and including PFNA than for the other studied PFASs. To our knowledge, this is the first report establishing relationships between PPARα activity and cellular concentration of a broad range of PFASs.

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The in vitro mutagenic and genotoxic potential of Heated Tobacco Products (HTPs) has already been studied with the particulate phase and reported previously. This study has been designed to complement the in vitro assessment of the HTP and to determine whether the inclusion of potential flavourings would alter the in vitro response by testing the other phase of the aerosol, the gas-vapour phase (GVP). Both flavoured and unflavoured Neostik GVP samples did not show any sign of mutagenic activity in the Ames test but induced a mutagenic response in the mouse lymphoma assay (MLA), however, these responses were significantly less than those of the reference cigarette, 3R4F. The results demonstrated that GVP emissions of this HTP did not induce either new qualitative or quantitative mutagenic hazards compared to 3R4F, as assessed by the Ames test (no new responsive strains) and MLA (a lower mutagenic response), respectively. A statistical comparative analysis of the responses showed that the addition of flavourings that may thermally decompose under the conditions of use did not add to the in vitro baseline responses of the unflavoured Neostik.

Background: Pesticide residue contamination of surface water in agricultural areas can have adverse effects on the ecosystem. We have performed an integrated chemical and bioanalytical profiling of surface water samples from Swedish agricultural areas, aiming to assess toxic activity due to presence of pesticides. A total of 157 water samples were collected from six geographical sites with extensive agricultural activity. The samples were chemically analyzed for 129 commonly used pesticides and transformation products. Furthermore, the toxicity was investigated using in vitro bioassays in the water samples following liquid-liquid extraction. Endpoints included oxidative stress response (Nrf2 activity), estrogen receptor (ER) activity, and aryl hydrocarbon receptor (AhR) activity. The bioassays were performed with a final enrichment factor of 5 for the water samples. All bioassays were conducted at non-cytotoxic conditions. Results: A total of 51 pesticides and transformation products were detected in the water samples. Most of the compounds were herbicides, followed by fungicides, insecticides and transformation products. The highest total pesticide concentration in an individual sample was 39 µg/L, and the highest median total concentration at a sample site was 1.1 µg/L. The largest number of pesticides was 31 in a single sample. We found that 3% of the water samples induced oxidative stress response, 23% of the samples activated the estrogen receptor, and 77% of the samples activated the aryl hydrocarbon receptor. Using Spearman correlation coefficients, a statistically significant correlation was observed between AhR and ER activities, and AhR activity was strongly correlated with oxidative stress in samples with a high AhR activity. Statistically significant relationships were observed between bioactivities and individual pesticides, although the relationships are probably not causal, due to the low concentrations of pesticides. Co-occurrence of non-identified chemical pollutants and naturally occurring toxic compounds may be responsible for the induced bioactivities. Conclusions: This study demonstrated that integrated chemical analysis and bioanalysis can be performed in water samples following liquid/liquid extraction with a final enrichment factor of 5. AhR and ER activities were induced in water samples from agricultural areas. The activities were presumably not caused by the occurrence of pesticides, but induced by other anthropogenic and natural chemicals.