Investigating the interaction of uranium(VI) with diatoms and their bacterial community: A microscopic and spectroscopic study.

Diatoms and bacteria play a vital role in investigating the ecological effects of heavy metals in the environment. Despite separate studies on metal interactions with diatoms and bacteria, there is a significant gap in research regarding heavy metal interactions within a diatom-bacterium system, which closely mirrors natural conditions. In this study, we aim to address this gap by examining the interaction of uranium(VI) (U(VI)) with Achnanthidium saprophilum freshwater diatoms and their natural bacterial community, primarily consisting of four successfully isolated bacterial strains (Acidovorax facilis, Agrobacterium fabrum, Brevundimonas mediterranea, and Pseudomonas peli) from the diatom culture. Uranium (U) bio-association experiments were performed both on the xenic A. saprophilum culture and on the four bacterial isolates. Scanning electron microscopy and transmission electron microscopy coupled with spectrum imaging analysis based on energy-dispersive X-ray spectroscopy revealed a clear co-localization of U and phosphorus both on the surface and inside A. saprophilum diatoms and the associated bacterial cells. Time-resolved laser-induced fluorescence spectroscopy with parallel factor analysis identified similar U(VI) binding motifs both on A. saprophilum diatoms and the four bacterial isolates. This is the first work providing valuable microscopic and spectroscopic data on U localization and speciation within a diatom-bacterium system, demonstrating the contribution of the co-occurring bacteria to the overall interaction with U, a factor non-negligible for future modeling and assessment of radiological effects on living microorganisms.

Advances in the Chemistry of Astatine and Implications for the Development of Radiopharmaceuticals.

ConspectusAstatine (At) is the rarest on Earth of all naturally occurring elements, situated below iodine in the periodic table. While only short-lived isotopes (t1/2 ≤ 8.1 h) are known, 211At is the object of growing attention due to its emission of high-energy alpha particles. Such radiation is highly efficient to eradicate disseminated tumors, provided that the radionuclide is attached to a cancer-targeting molecule. The interest in applications of 211At in nuclear medicine translates into the increasing number of cyclotrons able to produce it. Yet, many challenges related to the minute amounts of available astatine are to be overcome in order to characterize its physical and chemical properties. This point is of paramount importance to develop synthetic strategies and solve the labeling instability in current approaches that limits the use of 211At-labeled radiopharmaceuticals. Despite its discovery in the 1940s, only the past decade has seen a significant rise in the understanding of astatine's basic chemical and radiochemical properties, thanks to the development of new analytical and computational tools.In this Account, we give a concise summary of recent advances in the determination of the physicochemical properties of astatine, putting in perspective the duality of this element which exhibits the characteristics both of a halogen and of a metal. Striking features were evidenced in the recent determination of its Pourbaix diagram such as the identification of stable cationic species, At+ and AtO+, contrasting with other halogens. Like metals, these species were shown to form complexes with anionic ligands and to exhibit a particular affinity for organic species bearing soft donor atoms. On the other hand, astatine shares many characteristics with other halogen elements. For instance, the At- species exists in water, but with the least range of EH-pH stability in the halogen series. Astatine can form molecular interactions through halogen bonding, and it was only recently identified as the strongest halogen-bond donor. This ability is nonetheless affected by relativistic effects, which translate to other peculiarities for this heavy element. For instance, the spin-orbit coupling boosts astatine's propensity to form charge-shift bonds, catching up with the behavior of the lightest halogens (fluorine, chlorine).All these new data have an impact on the development of radiolabeling strategies to turn 211At into radiopharmaceuticals. Inspired by the chemistry of iodine, the chemical approaches have sparsely evolved over the past decades and have long been limited to electrophilic halodemetalation reactions to form astatoaryl compounds. Conversely, recent developments have favored the use of the more stable At- species including the aromatic nucleophilic substitution with diaryliodonium salts or the copper-catalyzed halodeboronation of arylboron precursors. However, it is clear that new bonding modalities are necessary to improve the in vivo stability of 211At-labeled aryl compounds. The tools and data gathered over the past decade will contribute to instigate original strategies for overcoming the challenges offered by this enigmatic element. Alternatives to the C-At bond such as the B-At and the metal-At bonds are typical examples of exciting new axes of research.

Pourbaix Diagram of Astatine Revisited: Experimental Investigations.

The Pourbaix diagram of an element displays its stable chemical forms with respect to the redox potential and pH of the solution, whose knowledge is fundamental for understanding and anticipating the chemistry of the element in a specified solution. Unlike most halogens, the Pourbaix diagram in the aqueous phase for astatine (At, Z = 85) is still under construction. In particular, the predominant domains of two astatine species assumed to exist under alkaline conditions, At- and AtO(OH)2-, need to be refined. Through high-performance ion-exchange chromatography, electromobility measurements, and competition experiments, the existence of At- and AtO(OH)2- has been confirmed and the associated standard potential has been determined for the first time (0.86 ± 0.05 V vs the standard hydrogen electrode). On the basis of these results, a revised version of astatine's Pourbaix diagram is proposed, covering the three oxidation states of astatine that exist in the thermodynamic stability range of water: At(-I), At(I), and At(III) (as At-, At+, AtO+, AtO(OH), and AtO(OH)2-).

Delocalized relativistic effects, from the viewpoint of halogen bonding.

The ability of organic and inorganic compounds bearing both iodine and astatine atoms to form halogen-bond interactions is theoretically investigated. Upon inclusion of the relativistic spin-orbit interaction, the I-mediated halogen bonds are more affected than the At-mediated ones in many cases. This unusual outcome is disconnected from the behavior of iodine's electrons. The significant decrease of astatine electronegativity with the spin-orbit coupling triggers a redistribution of the electron density, which propagates relativistic effects toward the distant iodine atom. This mechanism can be controlled by introducing suitable substituents and, in particular, strengthened by taking advantage of electron-withdrawing inductive and mesomeric effects. Noticeable relativistic effects can actually be transferred to light atoms properties, e.g., the halogen-bond basicity of bridgehead carbon atoms doubled in propellane derivatives.

Astatine Facing Janus: Halogen Bonding vs. Charge-Shift Bonding.

The nature of halogen-bond interactions was scrutinized from the perspective of astatine, potentially the strongest halogen-bond donor atom. In addition to its remarkable electronic properties (e.g., its higher aromaticity compared to benzene), C6At6 can be involved as a halogen-bond donor and acceptor. Two-component relativistic calculations and quantum chemical topology analyses were performed on C6At6 and its complexes as well as on their iodinated analogues for comparative purposes. The relativistic spin-orbit interaction was used as a tool to disclose the bonding patterns and the mechanisms that contribute to halogen-bond interactions. Despite the stronger polarizability of astatine, halogen bonds formed by C6At6 can be comparable or weaker than those of C6I6. This unexpected finding comes from the charge-shift bonding character of the C-At bonds. Because charge-shift bonding is connected to the Pauli repulsion between the bonding σ electrons and the σ lone-pair of astatine, it weakens the astatine electrophilicity at its σ-hole (reducing the charge transfer contribution to halogen bonding). These two antinomic characters, charge-shift bonding and halogen bonding, can result in weaker At-mediated interactions than their iodinated counterparts.

Towards a Stronger Halogen Bond Involving Astatine: Unexpected Adduct with Bu3 PO Stabilized by Hydrogen Bonding.

The halogen bond is a powerful tool for the molecular design and pushing the limits of its strength is of major interest. Bearing the most potent halogen-bond donor atom, astatine monoiodide (AtI) was recently successfully probed [Nat. Chem. 2018, 10, 428-434]. In this work, we continue the exploration of adducts between AtI and Lewis bases with the tributylphosphine oxide (Bu3 PO) ligand, revealing the unexpected experimental occurrence of two distinct chemical species with 1:1 and 2:1 stoichiometries. The 1:1 Bu3 PO⋅⋅⋅AtI complex is found to exhibit the strongest astatine-mediated halogen bond so far (with a formation constant of 10(4.24±0.35) ). Quantum chemical calculations unveil the intriguing nature of the 2:1 2Bu3 PO⋅⋅⋅AtI adduct, involving a halogen bond between AtI and one Bu3 PO molecular unit plus CH⋅⋅⋅O hydrogen bonds chelating the second Bu3 PO unit.

On the Interplay between Charge-Shift Bonding and Halogen Bonding.

The nature of halogen-bond interactions has been analysed from the perspective of the astatine element, which is potentially the strongest halogen-bond donor. Relativistic quantum calculations on complexes formed between halide anions and a series of Y3 C-X (Y=F to X, X=I, At) halogen-bond donors disclosed unexpected trends, e. g., At3 C-At revealing a weaker donating ability than I3 C-I despite a stronger polarizability. All the observed peculiarities have their origin in a specific component of C-Y bonds: the charge-shift bonding. Descriptors of the Quantum Chemical Topology show that the halogen-bond strength can be quantitatively anticipated from the magnitude of charge-shift bonding operating in Y3 C-X. The charge-shift mechanism weakens the ability of the halogen atom X to engage in halogen bonds. This outcome provides rationales for outlier halogen-bond complexes, which are at variance with the consensus that the halogen-bond strength scales with the polarizability of the halogen atom.

Questioning the Affinity of Electrophilic Astatine for Sulfur-containing Compounds: Unexpected Bindings Revealed.

The affinity of AtO+ for around 20 model ligands (L), carrying functionalized oxygen, sulfur, and nitrogen atoms, has been assessed through a combined experimental and theoretical methodology. Significant equilibrium constants (KL ∼ 104) have been measured for sulfur-containing compounds, in agreement with the previously highlighted, relatively stable radiolabeling of SH-containing proteins with 211At. Conversely, no interaction occurs in the aqueous phase for their oxygenated counterparts, but higher affinities (KL > 106) have been determined for nitrogen-based ligands, including aromatic nitrogen heterocycles. The quantum mechanical calculations definitively ruled out any rationale based on either the metallic character of astatine or its guessed softness; the favored interactions all involve specifically the oxygen atom of AtO+, leading to the formation of covalent O-S or O-C single bonds.

Nickel Retention on Callovo-Oxfordian Clay: Applicability of Existing Adsorption Models for Dilute Systems to Real Compact Rock.

In a waste management context, predicting the mobility of contaminants is essential. A key issue entails assessing the applicability of current knowledge on adsorption processes to natural systems. Such is the focus herein for nickel in interaction with Callovo-Oxfordian (COx) clay rock, a formation selected in France for possible radioactive waste disposal. The challenge is to link predictive modeling results with the experimental data characterizing the behavior of the labile and naturally occurring Ni fraction by implementing a new simple method. Retention studies on compact systems serve to complete this work. Combined electron microprobe and laser ablation high-resolution inductively coupled plasma mass spectrometry data show that natural Ni (∼39 mg kg-1) is homogeneously distributed within the clay matrix, which corresponds to the main reservoir (∼70%). Data interpretation of desorption tests yields an in situ Kd value of ∼80 L kg-1 and a labile Ni amount of ∼5 mg kg-1, that is, ∼13% of the Ni inventory. Predictive modeling explains the sorption data in considering that only weak clay fraction sites take part in the adsorption. The role of the clay matrix in Ni retention is confirmed by analyzing the Ni-spiked compact COx samples, whereby an increase of the Ni content in the clay fraction is observed following the retention experiment.

Evidence for the Heaviest Expected Halide Species in Aqueous Solution, At-, by Electromobility Measurements.

At- (astatide) is commonly expected to be the heaviest halide in the halogen group. However, there is no proof for the existence of this -1 charged species. Furthermore, investigations with astatine are restricted by its specific radioactive properties, which entail working at ultratrace concentrations (typically less than 10-10 M). In this work, an especially built electromigration device is applied to obtain information about the charge/size ratio characterizing an ion in aqueous solution. An anionic At species is observed in reducing conditions. Moreover, we propose the first absolute mobility value for the astatine species in acidic reducing condition: (-8.26 ± 0.59) × 10-4 cm2·V-1·s-1. This value appears close to that of I- ((-8.30 ± 0.33) × 10-4 cm2·V-1·s-1), which is obtained by the same method. The similar absolute mobilities obtained for both ions are coherent with theoretical calculations indicating similar diffusion behaviors for At- and I-. This good agreement confirms the existence of the At- species.

Spin-orbit coupling as a probe to decipher halogen bonding.

The nature of halogen-bond interactions is scrutinized from the perspective of astatine, the heaviest halogen element. Potentially the strongest halogen-bond donor, its ability is shown to be deeply affected by relativistic effects and especially by the spin-orbit coupling. Complexes between a series of XY dihalogens (X, Y = At, I, Br, Cl and F) and ammonia are studied with two-component relativistic quantum calculations, revealing that the spin-orbit interaction leads to a weaker halogen-bond donating ability of the diastatine species with respect to diiodine. In addition, the donating ability of the lighter halogen elements, iodine and bromine, in the AtI and AtBr species is more decreased by the spin-orbit coupling than that of astatine. This can only be rationalized from the evolution of a charge-transfer descriptor, the local electrophilicity ω+S,max, determined for the pre-reactive XY species. Finally, the investigation of the spin-orbit coupling effects by means of quantum chemical topology methods allows us to unveil the connection between the astatine propensity to form charge-shift bonds and the astatine ability to engage in halogen bonds.

Scrutinizing "Invisible" astatine: A challenge for modern density functionals.

The main-group 6p elements did not receive much attention in the development of recent density functionals. In many cases it is still difficult to choose among the modern ones a relevant functional for various applications. Here, we illustrate the case of astatine species (At, Z = 85) and we report the first, and quite complete, benchmark study on several properties concerning such species. Insights on geometries, transition energies and thermodynamic properties of a set of 19 astatine species, for which reference experimental or theoretical data has been reported, are obtained with relativistic (two-component) density functional theory calculations. An extensive set of widely used functionals is employed. The hybrid meta-generalized gradient approximation (meta-GGA) PW6B95 functional is overall the best choice. It is worth noting that the range-separated HSE06 functional as well as the old and very popular B3LYP and PBE0 hybrid-GGAs appear to perform quite well too. Moreover, we found that astatine chemistry in solution can accurately be predicted using implicit solvent models, provided that specific parameters are used to build At cavities. © 2016 Wiley Periodicals, Inc.

Advances on the Determination of the Astatine Pourbaix Diagram: Predomination of AtO(OH)2 (-) over At(-) in Basic Conditions.

It is generally assumed that astatide (At(-) ) is the predominant astatine species in basic aqueous media. This assumption is questioned in non-complexing and non-reductive aqueous solutions by means of high-pressure anion-exchange chromatography. Contrary to what is usually believed, astatide is found to be a minor species at pH=11. A different species, which also bears a single negative charge, becomes predominant when the pH is increased beyond 7. Using competition experiments, an equilibrium constant value of 10(-6.9) has been determined for the formation of this species from AtO(OH) with the exchange of one proton. The identification of this species, AtO(OH)2 (-) , is achieved through relativistic quantum mechanical calculations, which rule out the significant formation of the AtO2 (-) species, while leading to a hydrolysis constant of AtO(OH) in excellent agreement with experiment when the AtO(OH)2 (-) species is considered. Beyond the completion of the Pourbaix diagram of astatine, this new information is of interest for the development of (211) At radiolabeling protocols.

Unraveling the hydration-induced ground-state change of AtO+ by relativistic and multiconfigurational wave-function-based methods.

The AtO+ cation is one of the main chemical forms that appear in the astatine Pourbaix diagram. This form can react with closed-shell species in solution, while in the gas phase, it has a spin-triplet ground spin-orbit-free (SOF) state. Spin-orbit coupling (SOC) mixes its MS = 0 component with the 1Σ+ singlet-spin component, while keeping an essentially-spin-triplet SOC ground-state. Therefore, it was suggested that AtO+ undergoes a hydration-induced ground-state change to explain its reactivity in solution with closed-shell species [J. Phys. Chem. B, 2013, 117, 5206-5211]. In this work, we track the nature of the low-lying SOF and SOC states when the hydration sphere of AtO+ is stepwise increased, using relativistic and multiconfigurational wave-function-based methods. This work clarifies previous studies by (i) giving additional arguments justifying a solvation-induced ground-state change in this system and (ii) clearly identifying for the first time the nature of the involved SOF and SOC many-electron states. Indeed, we find at the SOF level that AtO+ undergoes a ground-state reversal between 3Σ- and the closed-shell component of 1Δ, which leads to an essentially-spin-singlet and closed-shell SOC ground-state. This explains the observed reactivity of AtO+ with closed-shell species in solution.

The Heaviest Possible Ternary Trihalogen Species, IAtBr- , Evidenced in Aqueous Solution: An Experimental Performance Driven by Computations.

Evidencing new chemical species in solution is particularly challenging when one works at ultra-trace concentrations, as is likely to happen with radioelements such as astatine (Z=85). Herein, quantum mechanical calculations were used to predict the narrow experimental domain in which it is possible to detect the presence of an exotic ternary trihalogen anion, IAtBr- , and thus to guide a series of experiments. By analyzing the outcomes of competition experiments, we show that IAtBr- exists and can even predominate in aqueous solution. The equilibrium constant associated with the reaction At+ +I- +Br- ⇌IAtBr- was determined to be 107.5±0.2 , which is in fair agreement with that predicted by density functional theory (106.9 ). This system not only constitutes the very first example of a ternary trihalogen species that involves the element astatine but is also the first trihalogen species reported to predominate in solution. Moreover, we show that the oxidation number of At is zero in this species, as in the other molecules and anions that At+ can form with Cl- , Br- , and I- ligands.

Effective bond orders from two-step spin-orbit coupling approaches: the I2, At2, IO(+), and AtO(+) case studies.

The nature of chemical bonds in heavy main-group diatomics is discussed from the viewpoint of effective bond orders, which are computed from spin-orbit wave functions resulting from spin-orbit configuration interaction calculations. The reliability of the relativistic correlated wave functions obtained in such two-step spin-orbit coupling frameworks is assessed by benchmark studies of the spectroscopic constants with respect to either experimental data, or state-of-the-art fully relativistic correlated calculations. The I2, At2, IO(+), and AtO(+) species are considered, and differences and similarities between the astatine and iodine elements are highlighted. In particular, we demonstrate that spin-orbit coupling weakens the covalent character of the bond in At2 even more than electron correlation, making the consideration of spin-orbit coupling compulsory for discussing chemical bonding in heavy (6p) main group element systems.

In vivo stability of 211At-radiopharmaceuticals: on the impact of halogen bond formation.

211At, when coupled to a targeting agent, is one of the most promising radionuclides for therapeutic applications. The main labelling approach consists in the formation of astatoaryl compounds, which often show a lack of in vivo stability. The hypothesis that halogen bond (XB) interactions with protein functional groups initiate a deastatination mechanism is investigated through radiochemical experiments and DFT modelling. Several descriptors agree on the known mechanism of iodoaryl substrates dehalogenation by iodothyronine deiodinases, supporting the higher in vivo dehalogenation of N-succinimidyl 3-[211At]astatobenzoate (SAB) conjugates in comparison with their iodinated counterparts. The guanidinium group in 3-[211At]astato-4-guanidinomethylbenzoate (SAGMB) prevents the formation of At-mediated XBs with the selenocysteine active site in iodothyronine deiodinases. The initial step of At-aryl bond dissociation is inhibited, elucidating the better in vivo stability of SAGMB conjugates compared with those of SAB. The impact of astatine's ability to form XB interactions on radiopharmaceutical degradation may not be limited to the case of aryl radiolabeling.

Radioactivity as a driver of bacterial community composition in naturally radioactive mineral springs in the French Massif Central.

Some natural environments on Earth are characterised by high levels of radiation, including naturally radioelement enriched mineral springs in the French Massif Central. Therefore, naturally radioactive mineral springs are interesting ecosystems for understanding how bacterial populations in these springs have adapted to high levels of natural and chronic radioactivity over the very long term. The aim of this study was to analyse the bacterial communities of sediments from five naturally radioactive mineral springs in the French Massif Central, sampled in autumn 2019 and spring 2020, and to observe whether radionuclides, compared to other physicochemical parameters, are drivers of the bacterial community structuring in these extreme environments. Physicochemical measurements showed that two springs, Dourioux and Montagne had high radioelement concentrations/activities (uranium, thorium and radon). Analysis of the structure of the bacterial communities, by next generation sequencing based on 16S rRNA gene sequencing, showed that the presence of radionuclides in Dourioux and Montagne, did not lead to a reduction in bacterial diversity and richness compared to the other springs. However, Dourioux and Montagne were characterised by specific bacterial populations, whose presence correlates with the radioelement concentrations/activities measured in these springs. This suggests that radioelements could partly explain the structuring of bacterial communities in these springs. In addition, several of these operational taxonomic units (OTUs) specific to Dourioux and Montagne, mainly affiliated to Proteobacteria, Firmicutes, Acidobacteria, Actinobacteria, and Bacteroidetes, could be involved in the biogeochemistry of radionuclides through different mechanisms (biosorption, biomineralisation, bioaccumulation, and bioreduction), which would allow the development of other bacterial species sensitive to these metals/radioelements. In particular, the co-occurrence of sulphate and/or iron-reducing bacteria, capable of bioreducing uranium, with fermentative bacteria, releasing sources of organic carbons, reflects associations of bacteria with complementary functions that allow them to grow in this peculiar environment and maintain a high diversity in these extreme environments. This study has provided a better understanding of the structuring of bacterial communities exposed to ionising radiation for thousands of years in naturally radioactive environments.

Assessing the chronic effect of the bioavailable fractions of radionuclides and heavy metals on stream microbial communities: A case study at the Rophin mining site.

This study aimed to assess the potential impact of long-term chronic exposure (69 years) to naturally-occurring radionuclides (RNs) and heavy metals on microbial communities in sediment from a stream flowing through a watershed impacted by an ancient mining site (Rophin, France). Four sediment samples were collected along a radioactivity gradient (for 238U368 to 1710 Bq.Kg-1) characterized for the presence of the bioavailable fractions of radionuclides (226Ra, 210Po), and trace metal elements (Th, U, As, Pb, Cu, Zn, Fe). Results revealed that the available fraction of contaminants was significant although it varied considerably from one element to another (0 % for As and Th, 5-59 % for U). Nonetheless, microbial communities appeared significantly affected by such chronic exposure to (radio)toxicities. Several microbial functions carried by bacteria and related with carbon and nitrogen cycling have been impaired. The high values of fungal diversity and richness observed with increasing downstream contamination (H' = 4.4 and Chao1 = 863) suggest that the community had likely shifted toward a more adapted/tolerant one as evidenced, for example, by the presence of the species Thelephora sp. and Tomentella sp. The bacterial composition was also affected by the contaminants with enrichment in Myxococcales, Acidovorax or Nostocales at the most contaminated points. Changes in microbial composition and functional structure were directly related to radionuclide and heavy metal contaminations, but also to organic matter which also significantly affected, directly or indirectly, bacterial and fungal compositions. Although it was not possible to distinguish the specific effects of RNs from heavy metals on microbial communities, it is essential to continue studies considering the available fraction of elements, which is the only one able to interact with microorganisms.

Unveiling the origins and transport processes of radioactive pollutants downstream from a former U-mine site using isotopic tracers and U-238 series disequilibrium.

High U concentrations (reaching up to 14,850 mg â‹… kg-1), were determined in soils and sediments of a wetland downstream of a former U mine in France. This study aims to identify the origin of radioactive contaminants in the wetland by employing Pb isotope fingerprinting, (234U/238U) disequilibrium, SEM, and SIMS observations. Additionally, information about U and 226Ra transport processes was studied using U-238 series disequilibrium. The results of Pb fingerprinting highlighted inherited material inputs of different U-mines with mainly two types of U-ores: i) pitchblende (UO2), and ii) parsonsite (Pb2(UO2)(PO4)2). Moreover, significant disequilibrium of (230Th/238U) and (226Ra/230Th) activity ratios highlighted the mobility of 238U and 226Ra in the wetland, primarily driven by the water table fluctuations. Finally, this work uncovered a limitation of Pb isotope fingerprinting in the case of parsonsite materials, as the high natural Pb content of this mineral may hide the uranogenic Pb signature in the samples.