Redox Reactivity of Cerium Oxide Nanoparticles Induces the Formation of Disulfide Bridges in Thiol-Containing Biomolecules.

The redox state of disulfide bonds is implicated in many redox control systems, such as the cysteine-cystine couple. Among proteins, ubiquitous cysteine-rich metallothioneins possess thiolate metal binding groups susceptible to metal exchange in detoxification processes. CeO2 NPs are commonly used in various industrial applications due to their redox properties. These redox properties that enable dual oxidation states (Ce(IV)/Ce(III)) to exist at their surface may act as oxidants for biomolecules. The interaction among metallothioneins, cysteine, and CeO2 NPs was investigated through various biophysical approaches to shed light on the potential effects of the Ce(4+)/Ce(3+) redox system on the thiol groups of these biomolecules. The possible reaction mechanisms include the formation of a disulfide bridge/Ce(III) complex resulting from the interaction between Ce(IV) and the thiol groups, leading to metal unloading from the MTs, depending on their metal content and cluster type. The formation of stable Ce(3+) disulfide complexes has been demonstrated via their fluorescence properties. This work provides the first evidence of thiol concentration-dependent catalytic oxidation mechanisms between pristine CeO2 NPs and thiol-containing biomolecules.

Osmoregulatory responses to cadmium in reference and historically metal contaminated Gammarus fossarum (Crustacea, Amphipoda) populations.

In order to better understand the variable sensitivities of crustaceans to metals, we investigated the impact of cadmium exposure in 3 populations of Gammarus fossarum from different rivers of France. The first population lives in a Cd-contaminated river from a geochemical background, while the others inhabit Cd-free sites. Osmoregulation, a relevant biomarker to evaluate crustacean health following metal contamination, was used as a proxy to evaluate the intra- and inter-populationnal sensitivities to Cd. Specimens from each population were experimentally exposed to 9 µg Cd2+/L Cd for 7 days and hemolymph osmolality (HO) was then individually measured. In exposed populations, high inter-individual variations in HO values were noted, resulting in their separation into non-impacted and slightly or highly Cd-impacted (with lower HO) animals. In gills of impacted organisms, deep histopathological alterations and protein overexpression of Na+/K+-ATPase and V-H+-ATPase were observed through histology and immunolocalization, while non-impacted animals showed profiles comparable to controls. Moreover, the osmoregulatory processes in the population living in the Cd-contaminated site were impacted by acute Cd exposure in the laboratory as much as for one of the two populations originating from Cd-free sites. The observed changes did not reveal any obvious adaptive osmoregulatory phenomena at the population scale, but they may be due to differences in fitness between individuals and between populations in relation to the features of their respective environments, unrelated with the presence of the metal.

In vivo-folded metal-metallothionein 3 complexes reveal the Cu-thionein rather than Zn-thionein character of this brain-specific mammalian metallothionein.

Metallothionein-3 (MT3) is one of the four mammalian metallothioneins (MT), and is constitutively synthesized in the brain. MT3 acts both intracellularly and extracellularly in this organ, performing functions related to neuronal growth and physiological metal (Zn and Cu) handling. It appears to be involved in the prevention of neurodegenerative disorders caused by insoluble Cu-peptide aggregates, as it triggers a Zn-Cu swap that may counteract the deleterious presence of copper in neural tissues. The literature data on MT3 coordination come from studies either on apo-MT3 reconstitution or the reaction of Zn-MT3 with Cu(2+) , an ion that is hardly present inside cells. To ascertain the MT3 metal-binding features in a scenario closer to the reductive cell cytoplasm, a study of the recombinant Zn(2+) , Cd(2+) and Cu(+) complexes of MT3, ßMT3, and αMT3, as well as the in vitro Zn(2+) -Cd(2+) and Zn(2+) -Cu(+) replacement processes, is presented here. We conclude that MT3 has a Cu-thionein character that is stronger than that of the MT1 and MT2 isoforms - also present in the mammalian brain - which is mainly contributed by its ß domain. In contrast, the α domain retains a high capacity to bind Zn(2+) ions, and, consequently, the entire MT3 peptide shows a peculiar dual ability to handle both metal ions. The nature of the formed Cu(+) -MT3 complexes oscillates from heterometallic Cu6 Zn4 -MT3 to homometallic Cu10 -MT3 major species, in a narrow Cu concentration range. Therefore, the entire MT3 peptide shows a high capacity to bind Cu(+) , provided that this occurs in a nonoxidative milieux. This reflects a peculiar property of this MT isoform, which accurately senses different Cu contents in the environment in which it is synthesized.

An adaptable mesocosm platform for performing integrated assessments of nanomaterial risk in complex environmental systems.

Physical-chemists, (micro)biologists, and ecologists need to conduct meaningful experiments to study the environmental risk of engineered nanomaterials with access to relevant mechanistic data across several spatial and temporal scales. Indoor aquatic mesocosms (60L) that can be tailored to virtually mimic any ecosystem appear as a particularly well-suited device. Here, this concept is illustrated by a pilot study aimed at assessing the distribution of a CeO2-based nanomaterial within our system at low concentration (1.5 mg/L). Physico-chemical as well as microbiological parameters took two weeks to equilibrate. These parameters were found to be reproducible across the 9-mesocosm setup over a 45-day period of time. Recovery mass balances of 115 ± 18% and 60 ± 30% of the Ce were obtained for the pulse dosing and the chronic dosing, respectively. This demonstrated the relevance of our experimental approach that allows for adequately monitoring the fate and impact of a given nanomaterial.

Mammalian MT1 and MT2 metallothioneins differ in their metal binding abilities.

Metallothioneins (MTs) constitute a universal family of polymorphic, ubiquitous small Cys-rich metal-binding polypeptides that in mammals are represented by four highly similar isoforms (MT1 to MT4). MT1 and MT2 have generally been considered as equivalent proteins, so that they are commonly referred to as MT1/MT2. However, transcription data have suggested a differential behavior for both gene products. In the present study, the metal binding abilities of mouse MT2 (mMT2) with divalent (Zn(II) and Cd(II)) and monovalent (Cu(I)) ions were analyzed and compared to those of the mouse MT1 (mMT1) isoform, previously determined using the same methodological approach. The comprehensive consideration of all the results obtained in this work experimentally demonstrates that the mMT2 isoform exhibits metal ion binding abilities distinct from those of mMT1, with a clear preference for Zn(II) coordination, if compared to Cu(I) or even to Cd(II). This is in full agreement with the gene expression regulation pattern for the MT1 and MT2 genes, as well as with the hypothesized preferential role of mMT2 in Zn(II) homeostasis mechanisms, while MT1, possibly differentiated from a most recent duplication event in the mammalian MT gene cluster, would have evolved to detoxify Cd(II), and probably other divalent metal ions.

Exposure to cerium dioxide nanoparticles differently affect swimming performance and survival in two daphnid species.

The CeO2 NPs are increasingly used in industry but the environmental release of these NPs and their subsequent behavior and biological effects are currently unclear. This study evaluates for the first time the effects of CeO2 NPs on the survival and the swimming performance of two cladoceran species, Daphnia similis and Daphnia pulex after 1, 10 and 100 mg.L⁻¹ CeO2 exposures for 48 h. Acute toxicity bioassays were performed to determine EC50 of exposed daphnids. Video-recorded swimming behavior of both daphnids was used to measure swimming speeds after various exposures to aggregated CeO2 NPs. The acute ecotoxicity showed that D. similis is 350 times more sensitive to CeO2 NPs than D. pulex, showing 48-h EC50 of 0.26 mg.L⁻¹ and 91.79 mg.L⁻¹, respectively. Both species interacted with CeO2 NPs (adsorption), but much more strongly in the case of D. similis. Swimming velocities (SV) were differently and significantly affected by CeO2 NPs for both species. A 48-h exposure to 1 mg.L⁻¹ induced a decrease of 30% and 40% of the SV in D. pulex and D. similis, respectively. However at higher concentrations, the SV of D. similis was more impacted (60% off for 10 mg.L⁻¹ and 100 mg.L⁻¹) than the one of D. pulex. These interspecific toxic effects of CeO2 NPs are explained by morphological variations such as the presence of reliefs on the cuticle and a longer distal spine in D. similis acting as traps for the CeO2 aggregates. In addition, D. similis has a mean SV double that of D. pulex and thus initially collides with twice more NPs aggregates. The ecotoxicological consequences on the behavior and physiology of a CeO2 NPs exposure in daphnids are discussed.