An assessment of the utility of the yeast GreenScreen assay in pharmaceutical screening.

In this paper we describe an initial reproducibility study of 12 proprietary compounds followed by the assessment of 51 marketed pharmaceuticals and, lastly, a summary of the data so far from 2698 proprietary compounds from the Johnson & Johnson (J&J) compound library, in the yeast GreenScreen assay (GSA). In this assay, a reporter system in the yeast cells employs the DNA damage inducible promoter of the RAD54 gene, fused to the extremely stable green fluorescent protein (GFP). The assay proved to be very robust, the Excel templates provided by Gentronix with the assay interfaced well with in-house J&J systems with little adaptation, the assay was very rapid to perform and used very little compound. The results confirm previous work which suggests that the yeast GSA detects different classes of genotoxic compounds to the Ames assay and as a result can help screen out important genotoxic compounds at the pre-regulatory test phase that are missed by Ames-test-based screens alone. A combination of SAR evaluation of genotoxicity plus an Ames-test-based screen and the GSA provides a powerful pre-regulatory test battery to aid in the selection of successful drug candidates.

Selenium bioaccumulation by the water boatman Trichocorixa reticulata (Guerin-Meneville).

The input of selenium from subsurface agricultural drainage into surface water systems can result in the accumulation of toxic concentrations of selenium in aquatic food chains. Elevated selenium concentrations in aquatic systems is a significant environmental problem in many areas of the United States. A laboratory investigation was conducted to determine the dominant route of selenium bioaccumulation by the corixid Trichocorixa reticulata, an important food chain organism. The roles of waterborne and foodborne exposure in selenium bioaccumulation were examined using 48-h bioassays. Waterborne selenium concentrations ranged from 0 to 1,000 microgram Se/L as selenate. A mixture of two species of blue-green algae cultured in media with selenium concentrations ranging from 0 to 1,000 microgram Se/L as selenate was used as a corixid diet in the foodborne treatments. Corixids exposed to waterborne selenate did not accumulate selenium above control concentrations. Corixids fed algae exposed to >/=100 microgram Se/L as selenate had significantly higher selenium concentrations than control organisms. These data suggest that corixids may be effectively isolated from the water and selenium accumulation is solely through dietary exposure.

Ecotoxicology of selenium in freshwater systems.

The toxicology, environmental impacts and risk assessment of Se in freshwater systems are a high priority for research and regulatory agencies. However, understanding Se in freshwater systems is a challenging endeavor. The accurate risk assessment and determination of a water-quality criterion for any freshwater ecosystem are difficult for many reasons. First, the understanding of the structure and energy dynamics in ecosystems is limited. Second, knowledge of Se cycling and transformations in aquatic ecosystems is rudimentary. Third, the role of various environmental factors affecting its bioaccumulation, biotransformation, and toxicology in aquatic ecosystems is largely unknown. Fourth, the extrapolation and use of existing data in commonly used formulas for risk assessment and calculations of water quality criteria is difficult because of the demonstrated species differences in the bioaccumulation, metabolism, and tolerance of Se. There are many aspects of Se ecotoxicology that need to be addressed to develop more accurate environmental risk assessments and determine appropriate water-quality criteria to protect aquatic ecosystems. Studies evaluating the biochemical speciation of Se in aquatic ecosystems, and determination of the role of microbial communities in its cycling, bioaccumulation, transformation, transfer, and toxicity in aquatic systems, appear to be priorities for future research. These would include a broad exploration of Se effects on ecosystems, e.g., exposure regime, direct and indirect biologic effects, and ecosystem level effects. There are, however, ecological uncertainties that tend to confound such endeavors, e.g., insufficient data, extrapolation issues, and environmental stochasticity (Harwell and Harwell 1989). There are several concepts concerning the ecotoxicology of Se that can be stated. Elevated concentrations have degraded many freshwater ecosystems throughout the United States, and additional systems are expected to be affected as anthropogenic activities increasingly mobilize Se into aquatic systems. Se is a very toxic essential trace element. Toxic threshold concentrations in water, dietary items, and tissues are only 2-5 times normal background concentrations. Toxicity in freshwater ecosystems is the result of bioaccumulation, biotransformation, and cycling of Se in aquatic food chains. Thus, environmental risk assessment and development of effective water-quality criteria to protect freshwater ecosystems become formidable tasks.

Effects of elevated foodborne selenium on growth and reproduction of the fathead minnow (Pimephales promelas).

Several field studies of selenium-contaminated lakes and reservoirs have indicated the possibility of selenium-induced reproductive failure in important populations of fish. These investigators have hypothesized that bioaccumulation of selenium through the food chain led to fish selenium levels high enough to elicit toxic responses. The present investigation was designed to determine the effects of elevated foodborne selenium on the fathead minnow (Pimephales promelas). Fish were fed a diet spiked with a mixture of inorganic (selenite and selenate) and organic (seleno-L-methionine) selenium and effects on growth and reproduction were determined. Growth was significantly inhibited at the highest selenium treatment levels evaluated (20 and 30 ppm Se). There were no significant treatment effects on any of the reproductive parameters measured. Reasons for the disparity between selenium-induced reproductive impairment observed in other species and apparent lack of impairment in fathead minnows may involve reduced bioaccumulation of selenium by minnows due to differences in gut morphology and physiology.