Non-target and suspect-screening analyses of hydroponic soybeans and passive samplers exposed to different watershed irrigation sources.

Water scarcity increases the likelihood of irrigating food crops with municipal wastewater that may pose potential dietary risks of regulated and non-regulated organic chemical uptake to edible plant tissues. Only a few studies have used high resolution mass spectrometry (HRMS) to assess the uptake of chemicals of concern into food crops. This study used non-target and suspect-screening analyses to compare total chemical features, tentatively identified chemicals (TICs), and EPA ToxCast chemicals in soybean plants and passive samplers exposed to five different irrigation sources that were collected from an agricultural watershed during mild drought conditions. Secondary-treated municipal wastewater effluent, two surface waters, two ground waters, and deionized municipal tap water were used for two hydroponic experiments: soybean roots and shoots and Composite Integrative Passive Samplers (CIPS) harvested after fourteen days of exposure and soybeans after fifty-six days. CIPS were sealed in separate glass amber jars to evaluate their efficacy to mimic chemical features, TICs, and ToxCast chemical uptake in plant roots, shoots, and beans. Total soybean biomass and water use were greatest for tap water, municipal wastewater, and surface water downstream of the municipal wastewater facility relative to groundwater samples and surface water collected upstream of the wastewater facility. ToxCast chemicals were ubiquitous across watershed irrigation sources in abundance, chemical use category, and number. Wastewater-exposed soybeans had the fewest extractable TICs in plant tissues of all irrigation sources. More ToxCast chemicals were identified in CIPS than extracted from irrigation sources by solid phase extraction. ToxCast chemicals in beans and CIPS were similar in number, chemical use category, and log Kow range. CIPS appear to serve as a useful surrogate for ToxCast chemical uptake in beans, the edible food product.

DNA-dependent protein kinase (DNA-PK) permits vascular smooth muscle cell proliferation through phosphorylation of the orphan nuclear receptor NOR1.

AIMS: Being central part of the DNA repair machinery, DNA-dependent protein kinase (DNA-PK) seems to be involved in other signalling processes, as well. NOR1 is a member of the NR4A subfamily of nuclear receptors, which plays a central role in vascular smooth muscle cell (SMC) proliferation and in vascular proliferative processes. We determined putative phosphorylation sites of NDA-PK in NOR1 and hypothesized that the enzyme is able to modulate NOR1 signalling and, this way, proliferation of SMC. METHODS AND RESULTS: Cultured human aortic SMC were treated with the specific DNA-PK inhibitor NU7026 (or siRNA), which resulted in a 70% inhibition of FCS-induced proliferation as measured by BrdU incorporation. Furthermore, FCS-stimulated up-regulation of NOR1 protein as well as the cell-cycle promoting proteins proliferating cell nuclear antigen (PCNA), cyclin D1, and hyperphosphorylation of the retinoblastoma protein were prevented by DNA-PK inhibition. Co-immunoprecipitation studies from VSM cell lysates demonstrated that DNA-PK forms a complex with NOR1. Mutational analysis and kinase assays demonstrated that NOR1 is a substrate of DNA-PK and is phosphorylated in the N-terminal domain. Phosphorylation resulted in post-transcriptional stabilization of the protein through prevention of its ubiquitination. Active DNA-PK and NOR1 were found predominantly expressed within the neointima of human atherosclerotic tissue specimens. In mice, inhibition of DNA-PK significantly attenuated neointimal lesion size 3 weeks after wire-injury. CONCLUSION: DNA-PK directly phosphorylates NOR-1 and, this way, modulates SMC proliferation. These data add to our understanding of vascular remodelling processes and opens new avenues for treatment of vascular proliferative diseases.