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Determining optimal conditions for the production of well diffracting crystals is a key step in every biocrystallography project. Here, a microfluidic device is described that enables the production of crystals by counter-diffusion and their direct on-chip analysis by serial crystallography at room temperature. Nine 'non-model' and diverse biomacromolecules, including seven soluble proteins, a membrane protein and an RNA duplex, were crystallized and treated on-chip with a variety of standard techniques including micro-seeding, crystal soaking with ligands and crystal detection by fluorescence. Furthermore, the crystal structures of four proteins and an RNA were determined based on serial data collected on four synchrotron beamlines, demonstrating the general applicability of this multipurpose chip concept., The following funding is acknowledged: Agence Nationale de la Recherche (contract No. ANR-11-LABX-0057_MITOCROSS to Claude Sauter, Bernard Lorber; contract No. ANR-10-LABX-0036_NETRN to Claude Sauter, Bernard Lorber; contract No. ANR-13-BS07-0007-01 to Eric Girard, Sylvain Engilberge); Ministère des Affaires Etrangères (contract No. PROCOPE Hubert Curien to Claude Sauter, Mario Mörl); Deutsche Forschungsgemeinschaft (contract No. Mo 634/10-1 to Mario Mörl, Heike Betat); Université de Strasbourg [grant No. Initiative d'excellence (IDEX) to Claude Sauter, Raphaël de Wijn]; Centre National de la Recherche Scientifique (grant No. MRCT-2012_PTI_UPR9002 to Claude Sauter).

[EN] The search for signs of life in the ancient rock record, extreme terrestrial environments, and other planetary bodiesrequires a well-established, universal, and unambiguous test of biogenicity. This is notably true for cellular remnantsof microbial life, since their relatively simple morphologies resemble various abiogenic microstructures that occur innature. Although lists of qualitative biogenicity criteria have been devised, debates regarding the biogenicity of manyancient microfossils persist to this day. We propose here an alternative quantitative approach for assessing thebiogenicity of putative microfossils. In this theoretical approach, different hypotheses—involving biology or not anddepending on the geologic setting—are put forward to explain the observed objects. These hypotheses correspond tospecific types of microstructures/systems. Using test samples, the morphology and/or chemistry of these systems arethen characterized at the scale of populations. Morphologic parameters include, for example, circularity, aspect ratio,and solidity, while chemical parameters could include elementary ratios (e.g., N/C ratio), isotopic enrichments (e.g.,d13C), or chirality (e.g., molar proportion of stereoisomers), among others. Statistic trends distinguishing the differentsystems are then searched for empirically. The trends found are translated into ‘‘decision spaces’’ where the differentsystems are quantitatively discriminated and where the potential microfossil population can be located as a singlepoint. This approach, which is formulated here on a theoretical level, will solve several problems associated with theclassical qualitative criteria of biogenicity. Most importantly, it could be applied to reveal the existence of cellular lifeon other planets, for which characteristics of morphology and chemical composition are difficult to predict., The Ministerio de Economia y competividad of Spain for funding the project CLG2016-78971-P. This project has received funding from the EuropeanResearch Council, under the European Union’s Horizon2020 research and innovation programme (grant agreementno. 694894) and the Seventh Framework Programme-FP7/2007-2013 (grant agreement no. 340863)

Bio compatible nanosystems exhibiting long lifetime ( ~millisecond) luminescence features are particu l arly relevant in the field of bioimaging. In this study, citrate functionalized calcium doped europium phosphates nanophosphors of the rhabdophane type were prepared at different synthesis times by a bioinspired crystallization route, consisting in thermal decomplexing of ca2•tEu3• {citrate{phosphate{car bonate solutions. The general formula of this material is Ca«Eu1 a(PO4) 1 a(HP04 l,,•nH2O, with CJ. ranging from 0 to 0.58 and n ~ 1. A thorough characterization of the nanoparticles has been carried out by XRD (including data processing with Topas 6.0), HR TEM, TEM, FTIR, TG{ITTA. ICP, dynamic light scattering (OLS), electrophoretic mobility, and fluorescence spectroscopy. Based on these results a crystallization mechanism involving the filling of cationic sites with Ca21ons associated to a concomitant adjustment of the P04{HPO4 ratio was proposed Upon calcium doping, the aspect ratio of the nanoparticles as well as of the crystalline domains decreased and the relative luminescence intensity (R.LI.) could be modulated. Neither the pH nor the ionic strength, nor the temperature (from 25 to 37 C) affected signif icantly the R.L.I. of particles after resuspension in water, leading to rather steady luminescence features usable in a large domain of conditions. This new class of luminescent compounds has been proved to be fully cytocompatible relative to GTL 16 human carcinoma cells and showed an improved cytocompatibil ity as the Ca2+ content increased when contacted with the more sensitive m17. ASC murine mesenchymal stem cells. These biocompatible nanoparticles thus appear as promising new tailorable tools for biomed ical applications as luminescent nanoprobes., We greatly acknowledge the project Biomin nanoapatite MAT2014 60533 R supported by Spanish MINEICO and co funded by FEDER and the Excellence Network of Crystallography and Crystallization "Factoría de Cristalización" FIS2015 71928 REDC supported by Spanish MINEICO. Cristóbal Verdugo Escamilla also acknowledges the Spanish MINEICO for his contract PT A2015 11103 I.

Nanomedicine covers the application of nanotechnologies in medicine. Of particular interest is the setup of highly-cytocompatible nanoparticles for use as drug carriers and/or for medical imaging. In this context, luminescent nanoparticles are appealing nanodevices with great potential for imaging of tumor or other targetable cells, and several strategies are under investigation. Biomimetic apatite nanoparticles represent candidates of choice in nanomedicine due to their high intrinsic biocompatibility and to the highly accommodative properties of the apatite structure, allowing many ionic substitutions. In this work, the preparation of biomimetic (bone-like) citrate-coated carbonated apatite nanoparticles doped with europium ions is explored using the citrate-based thermal decomplexing approach. The technique allows the preparation of the single apatitic phase with nanosized dimensions only at Eu doping concentrations ≤0.01 M at some timepoints. The presence of the citrate coating on the particle surface (as found in bone nanoapatites) and Eu substituting Ca is beneficial for the preparation of stable suspensions at physiological pH, as witnessed by the ζ-potential versus pH characterizations. The sensitized luminescence features of the solid particles, as a function of the Eu doping concentrations and the maturation times, have been thoroughly investigated, while those of particles in suspensions have been investigated at different pHs, ionic strengths and temperatures. Their cytocompatibility is illustrated in vitro on two selected cell types, the GTL-16 human carcinoma cells and the m17.ASC murine mesenchymal stem cells. This contribution shows the potentiality of the thermal decomplexing method for the setup of luminescent biomimetic apatite nanoprobes with controlled features for use in bioimaging., We greatly acknowledge the project Biomin-nanoapatite MAT2014-60533-R supported by Spanish MINEICO and co-funded by FEDER and the Excellence Network of Crystallography and Crystallization “Factoría de Cristalización” FIS2015-71928-REDC supported by Spanish MINEICO. Cristóbal Verdugo-Escamilla also acknowledges the Spanish MINEICO for his contract PTA2015-11103-I.

Crystallographic methods are used to identify the primary mineral phase of pseudomorphs of crystals embedded in 3.48 Ga bedded carbonate-chert rocks from the Dresser Formation, Pilbara Craton, Australia. This identification provides valuable information on the chemical environments at the onset of life on Earth., Crystallography has a long history of providing knowledge and methods for applications in other disciplines. The identification of minerals using X-ray diffraction is one of the most important contributions of crystallography to earth sciences. However, when the crystal itself has been dissolved, replaced or deeply modified during the geological history of the rocks, diffraction information is not available. Instead, the morphology of the crystal cast provides the only crystallographic information on the original mineral phase and the environment of crystal growth. This article reports an investigation of crystal pseudomorphs and crystal casts found in a carbonate-chert facies from the 3.48 Ga-old Dresser Formation (Pilbara Craton, Australia), considered to host some of the oldest remnants of life. A combination of X-ray microtomography, energy-dispersive X-ray spectroscopy and crystallographic methods has been used to reveal the original phases of these Archean pseudomorphs. It is found with a high degree of confidence that the original crystals forming in Archean times were hollow aragonite, the high-temperature polymorphs of calcium carbonate, rather than other possible alternatives such as gypsum (CaSO4·2H20) and nahcolite (NaHCO3). The methodology used is described in detail.

International audience; Nanocrystalline calcium phosphate apatites constitute the main inorganic part of hard tissues, and a growing focus is devoted to prepare synthetic analogs, so-called "biomimetic", able to precisely mimic the morphological and physico-chemical features of biological apatite compounds. Both from fundamental and applied viewpoints, an accurate characterization of nanocrystalline apatites, including their peculiar surface features, and a deep knowledge of crystallization aspects are prerequisites to attempt understanding mineralization phenomena in vivo as well as for designing innovative bioactive materials that may then find applications in bone tissue engineering, either as self-supported scaffolds and fillers or in the form of coatings, but also in other domains such as drug delivery or else medical imaging. Also,interfacial phenomena are of prime importance for getting a better insight of biomineralization and for following the behavior of biomaterials in or close to their final conditions of use. In this view,both adsorption and ion exchange represent essential processes involving the surface of apatite nanocrystals, possibly doped with foreign elements or functionalized with organic molecules of interest. In this review paper, we will address these various points in details based on a large literature survey. We will also underline the fundamental physico-chemical and behavioral differences that exist between nanocrystalline apatites (whether of biological origin or their synthetic biomimetic analogs) and stoichiometric hydroxyapatite.

International audience; Cancer vaccines based on virus-like particles (VLPs) vectors may offer many advantages over other antigen-delivery systems and represent an alternative to the ex vivo cell therapy approach. In this study, we describe the use of penton-dodecahedron (Pt-Dd) VLPs from human adenovirus type 3 (Ad3) as cancer vaccine vehicle for specific antigens, based on its unique cellular internalization properties. WW domains from the ubiquitin ligase Nedd4 serve as an adapter to bind the antigen to Pt-Dd. By engineering fusion partners of WW with the model antigen ovalbumin (OVA), Pt-Dd can efficiently deliver WW-OVA in vitro and the Pt-Dd/WW complex can be readily internalized by dendritic cells (DCs). Immunization with WW-OVA/Pt-Dd results in 90% protection against B16-OVA melanoma implantation in syngeneic mice. This high level of protection correlates with the development of OVA-specific CD8(+) T cells. Moreover, vaccination with WW-OVA Pt-Dd induces robust humoral responses in mice as shown by the high levels of anti-OVA antibodies (Abs) detected in serum. Importantly, treatment of mice bearing B16-OVA tumors with WW-OVA/Pt-Dd results in complete tumor regression in 100% of cases. Thus, our data supports a dual role of Pt-Dd as antigen-delivery vector and natural adjuvant, able to generate integrated cellular and humoral responses of broad immunogenic complexity to elicit specific antitumor immunity. Antigen delivery by Pt-Dd vector is a promising novel strategy for development of cancer vaccines with important clinical applications.