Chemical alternatives assessment of different flame retardants - A case study including multi-walled carbon nanotubes as synergist.

Flame retardants (FRs) are a diverse group of chemicals used as additives in a wide range of products to inhibit, suppress, or delay ignition and to prevent the spread of fire. Halogenated FRs (HFRs) are widely used because of their low impact on other material properties and the low loading levels necessary to meet the required flame retardancy. Health and environmental hazards associated with some halogenated FRs have driven research for identifying safer alternatives. A variety of halogen-free FRs are available on the market, including organic (phosphorus and nitrogen based chemicals) and inorganic (metals) materials. Multi-walled carbon nanotubes (MWCNT) have been demonstrated to act as an effective/synergistic co-additive in some FR applications and could thereby contribute to reducing the loading of FRs in products and improving their performance. As part of the FP7 project DEROCA we carried out a chemical alternatives assessment (CAA). This is a methodology for identifying, comparing and selecting safer alternatives to chemicals of concern based on criteria for categorising human and environmental toxicity as well as environmental fate. In the project we assessed the hazard data of different halogen-free FRs to be applied in 5 industrial and consumer products and here we present the results for MWCNT, aluminium diethylphosphinate, aluminium trihydroxide, N-alkoxy hindered amines and red phosphorus compared to the HFR decabromodiphenylether. We consulted the REACH guidance, the criteria of the U.S.-EPA Design for Environment (DfE) and the GreenScreen® Assessment to assess and compare intrinsic properties affecting the hazard potential. A comparison/ranking of exposure reference values such as Derived No Effect Levels (DNELs) showed that FRs of concern are not identified by a low DNEL. A comparison based on hazard designations according to the U.S.-EPA DfE and GreenScreen® for human health endpoints, aquatic toxicity and environmental fate showed that the major differences between FRs of concern and their proposed alternatives are the potential for bioaccumulation and CMR (carcinogenic, mutagenic or reprotoxic) effects. As most alternatives are inorganic chemicals, persistence (alone) is not a suitable criterion. From our experiences in carrying out a CAA we conclude: i) REACH registration dossiers provide a comprehensive source of hazard information for an alternative assessment. It is important to consider that the presented data is subject to changes and its quality is variable. ii) Correct identification of the chemicals is crucial to retrieve the right data. This can be challenging for mixtures, reaction products or nanomaterials or when only trade names are available. iii) The quality of the data and the practice on how to fill data gaps can have a huge impact on the results and conclusions. iv) Current assessment criteria have mainly been developed for organic chemicals and create challenges when applied to inorganic solids, including nanomaterials. It is therefore crucial to analyse and report uncertainties for each decision making step.

Genome-wide single-cell-level screen for protein abundance and localization changes in response to DNA damage in S. cerevisiae.

An effective response to DNA damaging agents involves modulating numerous facets of cellular homeostasis in addition to DNA repair and cell-cycle checkpoint pathways. Fluorescence microscopy-based imaging offers the opportunity to simultaneously interrogate changes in both protein level and subcellular localization in response to DNA damaging agents at the single-cell level. We report here results from screening the yeast Green Fluorescent Protein (GFP)-fusion library to investigate global cellular protein reorganization on exposure to the alkylating agent methyl methanesulfonate (MMS). Broad groups of induced, repressed, nucleus- and cytoplasm-enriched proteins were identified. Gene Ontology and interactome analyses revealed the underlying cellular processes. Transcription factor (TF) analysis identified principal regulators of the response, and targets of all major stress-responsive TFs were enriched amongst the induced proteins. An unexpected partitioning of biological function according to the number of TFs targeting individual genes was revealed. Finally, differential modulation of ribosomal proteins depending on methyl methanesulfonate dose was shown to correlate with cell growth and with the translocation of the Sfp1 TF. We conclude that cellular responses can navigate different routes according to the extent of damage, relying on both expression and localization changes of specific proteins.

Genomic phenotyping of the essential and non-essential yeast genome detects novel pathways for alkylation resistance.

BACKGROUND: A myriad of new chemicals has been introduced into our environment and exposure to these agents can damage cells and induce cytotoxicity through different mechanisms, including damaging DNA directly. Analysis of global transcriptional and phenotypic responses in the yeast S. cerevisiae provides means to identify pathways of damage recovery upon toxic exposure. RESULTS: Here we present a phenotypic screen of S. cerevisiae in liquid culture in a microtiter format. Detailed growth measurements were analyzed to reveal effects on ~5,500 different haploid strains that have either non-essential genes deleted or essential genes modified to generate unstable transcripts. The pattern of yeast mutants that are growth-inhibited (compared to WT cells) reveals the mechanisms ordinarily used to recover after damage. In addition to identifying previously-described DNA repair and cell cycle checkpoint deficient strains, we also identified new functional groups that profoundly affect MMS sensitivity, including RNA processing and telomere maintenance. CONCLUSIONS: We present here a data-driven method to reveal modes of toxicity of different agents that impair cellular growth. The results from this study complement previous genomic phenotyping studies as we have expanded the data to include essential genes and to provide detailed mutant growth analysis for each individual strain. This eukaryotic testing system could potentially be used to screen compounds for toxicity, to identify mechanisms of toxicity, and to reduce the need for animal testing.