The increase of production volumes of silver nanowires (AgNWs) and of consumer products incorporating them, may lead to increased health risks from occupational and public exposures. There is currently limited information about the putative toxicity of AgNWs upon inhalation, and incomplete understanding of the properties that control their bioreactivity. The lung lining fluid (LLF), which contains phospholipids and surfactant proteins, represents a first contact site with the respiratory system. In this work, the impact of Dipalmitoylphosphatidylcholine (DPPC), Curosurf® and murine LLF on the stability of AgNWs was examined. Both the phospholipid and protein components of the LLF modified the dissolution kinetics of AgNWs, due to the formation of a lipid corona or aggregation of the AgNWs. Moreover, the hydrophilic, but neither the hydrophobic surfactant proteins nor the phospholipids, induced agglomeration of the AgNWs. Finally, the generation of a secondary population of nano-silver was observed and attributed to the reduction of Ag+ ions by the surface capping of the AgNWs. Our findings highlight that combinations of spatially resolved dynamic and static techniques are required to develop a holistic understanding of which parameters govern AgNW behavior at the point of exposure and to accurately predict their risks on human health and the environment.

Fe-S cluster biogenesis is of interest to many fields, including bioenergetics and gene regulation. The CSD system is one of three Fe-S cluster biogenesis systems in E. coli and is comprised of the cysteine desulfurase CsdA, the sulfur acceptor protein CsdE, and the E1-like protein CsdL. The biological role, biochemical mechanism, and protein targets of the system remain uncharacterized. Here we present that the active site CsdE C61 has a lowered pK a value of 6.5, which is nearly identical to that of C51 in the homologous SufE protein and which is likely critical for its function. We observed that CsdE forms disulfide bonds with multiple proteins and identified the proteins that copurify with CsdE. The identification of Fe-S proteins and both putative and established Fe-S cluster assembly (ErpA, glutaredoxin-3, glutaredoxin-4) and sulfur trafficking (CsdL, YchN) proteins supports the two-pathway model, in which the CSD system is hypothesized to synthesize both Fe-S clusters and other sulfur-containing cofactors. We suggest that the identified Fe-S cluster assembly proteins may be the scaffold and/or shuttle proteins for the CSD system. By comparison with previous analysis of SufE, we demonstrate that there is some overlap in the CsdE and SufE interactomes. Keywordsiron-sulfur (Fe-S) cluster biogenesis; iron-sulfur cluster assembly; iron-sulfur cluster biosynthesis; cysteine sulfinate desulfinase (CSD); pK a ; CsdA; CsdE; CsdL; SufE Sulfur trafficking pathways are highly conserved, complex protein machineries that are responsible for extracting sulfur from cysteine and synthesizing the sulfur-containing cofactors and thionucleosides, including thiamin, biotin, the molybdenum cofactor, lipoic acid, S-adenosylmethionine, and iron-sulfur (Fe-S) clusters (reviewed in Mueller 2006) 1 . The first step of sulfur trafficking is removal of inorganic sulfur from free cysteine by a cysteine desulfurase, followed by the transient and covalent binding of the sulfur atom to a reactive cysteine residue sulfhydryl within the desulfurase active site, thereby forming a persulfide (R-S-SH) intermediate. Due to the toxicity of free sulfide, the persulfide intermediate is the universal sulfur donor in these pathways. The sulfur atom is then transferred between nucleophilic cysteine residue sulfhydryls on various protein carriers, * Author to whom correspondence should be addressed: Department of Environmental Toxicology University of California, Davis One Shields Ave 4138 Meyer Hall Davis, CA 95616 Phone: 530-754-2271 Fax: 530-752-3394. SUPPORTING INFORMATION AVAILABLE Table S1 (Fig. 3C), Table S2 (Fig. 4A), and Table S3 (Fig. 4A) contain proteomic parameters for the proteins identified. Table S1 ( Fig. 3C) contains accession numbers, molecular weight, protein score, number nonredundant peptides, protein score confidence interval, and sequence coverage. Table S2 (Fig. 4A) groups the proteins by function, presents protein molecular weight, and identifies the proteins containing Fe-S clusters. Table S3 (Fig. 4A) contains acc...

Multiple studies have examined the direct cellular toxicity of silver nanoparticles (AgNPs). However, the lung is a complex biological system with multiple cell types and a lipid-rich surface fluid; therefore, organ level responses may not depend on direct cellular toxicity. We hypothesized that interaction with the lung lining is a critical determinant of organ level responses. Here, we have examined the effects of low dose intratracheal instillation of AgNPs (0.05 µg/g body weight) 20 and 110nm diameter in size, and functionalized with citrate or polyvinylpyrrolidone. Both size and functionalization were significant factors in particle aggregation and lipid interaction in vitro. One day post-intratracheal instillation lung function was assessed, and bronchoalveolar lavage (BAL) and lung tissue collected. There were no signs of overt inflammation. There was no change in surfactant protein-B content in the BAL but there was loss of surfactant protein-D with polyvinylpyrrolidone (PVP)-stabilized particles. Mechanical impedance data demonstrated a significant increase in pulmonary elastance as compared to control, greatest with 110nm PVP-stabilized particles. Seven days post-instillation of PVP-stabilized particles increased BAL cell counts, and reduced lung function was observed. These changes resolved by 21 days. Hence, AgNP-mediated alterations in the lung lining and mechanical function resolve by 21 days. Larger particles and PVP stabilization produce the largest disruptions. These studies demonstrate that low dose AgNPs elicit deficits in both mechanical and innate immune defense function, suggesting that organ level toxicity should be considered.

A valuable approach to chemical safety assessment is the use of read-across chemicals to provide safety data to support the assessment of structurally similar chemicals. An inventory of over 6000 discrete organic chemicals used as fragrance materials in consumer products has been clustered into chemical class-based groups for efficient search of read-across sources. We developed a robust, tiered system for chemical classification based on (1) organic functional group, (2) structural similarity and reactivity features of the hydrocarbon skeletons, (3) predicted or experimentally verified Phase I and Phase II metabolism, and (4) expert pruning to consider these variables in the context of specific toxicity end points. The systematic combination of these data yielded clusters, which may be visualized as a top-down hierarchical clustering tree. In this tree, chemical classes are formed at the highest level according to organic functional groups. Each subsequent subcluster stemming from classes in this hierarchy of the cluster is a chemical cluster defined by common organic functional groups and close similarity in the hydrocarbon skeleton. By examining the available experimental data for a toxicological endpoint within each cluster, users can better identify potential read-across chemicals to support safety assessments.