Centre de Biophysique Moléculaire - UPR 4301

Critical to the origin of life are the ingredients of life, of course, but also the physical and chemical conditions in which prebiotic chemical reactions can take place. These factors place constraints on the types of Hadean environment in which life could have emerged. Many locations, ranging from hydrothermal vents and pumice rafts, through volcanic-hosted splash pools to continental springs and rivers, have been proposed for the emergence of life on Earth, each with respective advantages and certain disadvantages. However, there is another, hitherto unrecognized environment that, on the Hadean Earth (4.5–4.0 Ga), would have been more important than any other in terms of spatial and temporal scale : the sedimentary layer between oceanic crust and seawater. Using as an example sediments from the 3.5–3.33 Ga Barberton Greenstone Belt, South Africa, analogous at least on a local scale to those of the Hadean eon, we document constant permeation of the porous, carbonaceous, and reactive sedimentary layer by hydrothermal fluids emanating from the crust. This partially UV-protected, subaqueous sedimentary environment, characterized by physical and chemical gradients, represented a widespread system of miniature chemical reactors in which the production and complexification of prebiotic molecules could have led to the origin of life. Key Words : Origin of life-Hadean environment-Mineral surface reactions-Hydrothermal fluids-Archean volcanic sediments.

A concise outlook on the potential role of confinement in phosphorylation and phosphate condensation pertaining to prebiotic chemistry is presented. Inorganic confinement is a relatively uncharted domain in studies concerning prebiotic chemistry, and even more so in terms of experimentation. However, molecular crowding within confined dimensions is central to the functioning of contemporary biology. There are numerous advantages to confined environments and an attempt to highlight this fact, within this article, has been undertaken, keeping in context the limitations of aqueous phase chemistry in phosphorylation and, to a certain extent, traditional approaches in prebiotic chemistry.

Cell fixation is an essential approach for preserving cell morphology, allowing the targeting and labelling of biomolecules with fluorescent probes. One of the key requirements for more efficient fluorescent labelling is the preservation of cell morphology, which usually requires a combination of several fixation techniques. In addition, the use of a counter stain is often essential to improve the contrast of the fluorescent probes. Current agents possess significant limitations, such as low resistance toward photobleaching and sensitivity to changes in the microenvironment. Luminescent Ln3+ ‘encapsulated sandwich’ metallacrowns (MCs) overcome these drawbacks and offer complementary advantages. In particular, they emit sharp emission bands, possess a large difference between excitation and emission wavelengths and do not photobleach. Herein, MCs formed with pyrazinehydroxamic acid (Ln3+[Zn(II)MCpyzHA], Ln3+ = Yb, Nd) were used, combined with near-infrared (NIR) counter staining and fixation agents for HeLa cells upon an initial five minute exposure to UV-A light. The validity and quality of the cell fixation were assessed with Raman spectroscopy. Analysis of the NIR luminescence properties of these MCs was performed under different experimental conditions, including in a suspension of stained cells. Moreover, the high emission intensity of Ln3+[Zn(II)MCpyzHA] in the NIR region allows these MCs to be used for imaging with standard CCD cameras installed on routine fluorescence microscopes. Finally, the NIR-emitting Ln3+[Zn(II)MCpyzHA] compounds combine, within a single molecule, features such as cell fixation and staining abilities, good photostability and minimal sensitivity of the emission bands to the local microenvironment, and they are highly promising for establishing the next generation of imaging agents with a single biodistribution.

In this study, we attempt to illustrate the competition that constitutes the main challenge of astrobiology, namely the competition between the probability of extraterrestrial life and its detectability. To illustrate this fact, we propose a simple statistical approach based on our knowledge of the Universe and the Milky Way, the Solar System, and the evolution of life on Earth permitting us to obtain the order of magnitude of the distance between Earth and bodies inhabited by more or less evolved past or present life forms, and the consequences of this probability for the detection of associated biosignatures. We thus show that the probability of the existence of evolved extraterrestrial forms of life increases with distance from the Earth while, at the same time, the number of detectable biosignatures decreases due to technical and physical limitations. This approach allows us to easily explain to the general public why it is very improbable to detect a signal of extraterrestrial intelligence while it is justified to launch space probes dedicated to the search for microbial life in the Solar System.

The Close-Up Imager (CLUPI) onboard the ESA ExoMars Rover is a powerful high-resolution color camera specifically designed for close-up observations. Its accommodation on the movable drill allows multiple positioning. The science objectives of the instrument are geological characterization of rocks in terms of texture, structure, and color and the search for potential morphological biosignatures. We present the CLUPI science objectives, performance, and technical description, followed by a description of the instrument’s planned operations strategy during the mission on Mars. CLUPI will contribute to the rover mission by surveying the geological environment, acquiring close-up images of outcrops, observing the drilling area, inspecting the top portion of the drill borehole (and deposited fines), monitoring drilling operations, and imaging samples collected by the drill. A status of the current development and planned science validation activities is also given. Key Words : Mars ?Biosignatures ?Planetary Instrumentation. Astrobiology 17, 595 ?611.
The Close-Up Imager (CLUPI) onboard the ESA ExoMars Rover is a powerful high-resolution color camera specifically designed for close-up observations. Its accommodation on the movable drill allows multiple positioning. The science objectives of the instrument are geological characterization of rocks in terms of texture, structure, and color and the search for potential morphological biosignatures. We present the CLUPI science objectives, performance, and technical description, followed by a description of the instrument’s planned operations strategy during the mission on Mars. CLUPI will contribute to the rover mission by surveying the geological environment, acquiring close-up images of outcrops, observing the drilling area, inspecting the top portion of the drill borehole (and deposited fines), monitoring drilling operations, and imaging samples collected by the drill. A status of the current development and planned science validation activities is also given. Key Words : Mars ?Biosignatures ?Planetary Instrumentation. Astrobiology 17, 595 ?611.

Raman spectroscopy has undergone rapid development over the last few decades. The ability to acquire a spectrum in only a few tens of milliseconds allows use of Raman mapping as a routine technique. However, with respect to classical single spectrum measurement, this technique is not still as widely used as it could be, in particular for mineralogy and petrography. Here, we explain the advantages of Raman mapping for obtaining additional information compared to single spot analyses. The principle and the limits of the technique are first explained in 2D and 3D. Data processing techniques are then described using different types of rocks and minerals to demonstrate the utility of Raman mapping for obtaining information about the general composition, identification of small phases, as well as for distinguishing minerals that are spectrally very close. More “exotic” uses of the collected signal are also described. Finally, a gallery of images from representative samples is used to illustrate the discussion.

The SPectral Imager (SPIM) facility is a laboratory VIS-IR spectrometer developed to support spaceborne observations of rocky bodies of the solar system. Currently, this laboratory setup is used to support the Dawn NASA mission and to support the 2018 ExoMars mission in the spectral investigation of Martian subsurface. Specifically, for this mission, a selection of relevant Mars analogue materials has been characterized and stored in the International Space Analogue Rockstore (ISAR), hosted in Orléans, France. In this investigation, two volcanic rock samples from the ISAR collection were analyzed. These two samples were chosen because of their similarity in mineralogical composition and age with Martian basalts and volcanic sands. Moreover, volcanic sands are particularly interesting because they can contain fossils of primitive life forms. The analysis of data collected by SPIM resulted in good agreement with the mineralogical phases detected in these two samples by mineralogical and petrographical techniques, demonstrating the effectiveness of the high spatial and spectral resolution of SPIM for identifying and for mapping different mineralogical species on cut rock and mineral samples.

The co-delivery of minicircle DNA (mcDNA) and small anti-cancer drugs via stimuli-sensitive nanocarriers is a promising approach for combinatorial cancer therapy. However, the simultaneous loading of drugs and DNA in nanosized delivery systems is remarkably challenging. In this study we describe the synthesis of triblock copolymer micelles based on poly(2-ethyl-2-oxazoline)-poly(L-lactide) grafted with bioreducible polyethylenimine (PEOz-PLA-g-PEI-SS) for co-delivery of supercoiled (sc) mcDNA vectors and Doxorubicin (Dox). These amphiphilic carriers take advantage of non-fouling oxazolines to confer biological stability, of PLA to provide a hydrophobic core for drug encapsulation and of bioreducible PEI-SS to provide mcDNA complexation and an on-demand stimuli-responsive release. The obtained results show that mcDNA-loaded micelleplexes penetrate into in vitro tumor spheroid models with specific kinetics and exhibit a higher gene expression when compared to non-bioreducible nanocarriers. Moreover, in vivo bioluminescence imaging showed that gene expression is detected up to 8days following mcDNA-micelles intratumoral administration. Furthermore, drug-gene co-delivery in PEOz-PLA-g-PEI-SS carriers was verified by successful encapsulation of both Dox and mcDNA with high efficacy. Moreover, dual-loaded micelleplexes presented significant uptake and a cytotoxic effect in 2D cultures of cancer cells. The co-delivery of mcDNA-Dox to B16F10-Luciferase tumor bearing mice resulted in a reduction in tumor volume and cancer cells viability. Overall, such findings indicate that bioreducible triblock micelles are efficient for focal delivery in vivo and have potential for future application in combinatorial DNA-drug therapy.

The search for traces of life is one of the principal objectives of Mars exploration. Central to this objective is the concept of habitability, the set of conditions that allows the appearance of life and successful establishment of microorganisms in any one location. While environmental conditions may have been conducive to the appearance of life early in martian history, habitable conditions were always heterogeneous on a spatial scale and in a geological time frame. This "punctuated" scenario of habitability would have had important consequences for the evolution of martian life, as well as for the presence and preservation of traces of life at a specific landing site. We hypothesize that, given the lack of long-term, continuous habitability, if martian life developed, it was (and may still be) chemotrophic and anaerobic. Obtaining nutrition from the same kinds of sources as early terrestrial chemotrophic life and living in the same kinds of environments, the fossilized traces of the latter serve as useful proxies for understanding the potential distribution of martian chemotrophs and their fossilized traces. Thus, comparison with analog, anaerobic, volcanic terrestrial environments (Early Archean >3.5-3.33 Ga) shows that the fossil remains of chemotrophs in such environments were common, although sparsely distributed, except in the vicinity of hydrothermal activity where nutrients were readily available. Moreover, the traces of these kinds of microorganisms can be well preserved, provided that they are rapidly mineralized and that the sediments in which they occur are rapidly cemented. We evaluate the biogenicity of these signatures by comparing them to possible abiotic features. Finally, we discuss the implications of different scenarios for life on Mars for detection by in situ exploration, ranging from its non-appearance, through preserved traces of life, to the presence of living microorganisms. KEY WORDS : Mars-Early Earth-Anaerobic chemotrophs-Biosignatures-Astrobiology missions to Mars.

Interacting, diverse microbe-sediment systems exist in natural environments today but have not yet been recognized in the oldest records of life on Earth (older than 3.3 Ga) because of lack of distinctive biomarker molecules and patchy preservation of microbial paleocommunities. In an in-situ outcrop- to microbial-scale study, we have differentiated probable phototrophic, chemolithotrophic, and chemo-organotrophic fossil microbial signatures in a nearshore volcanogenic sedimentary setting in 3.33 Ga rocks of the Josefsdal Chert, Barberton greenstone belt, South Africa, while demonstrating the importance of contemporaneous hydrothermal activity. Hydrothermal fluids, as a nutrient source, strongly controlled the development and distribution of the microbial communities and, as a silicifying agent, contributed to their rapid fossilization. We thus show that intricate microbe-sediment systems are deep-rooted in time and that at least some early life may indeed have been thermophilic.

Demonstrating the biogenicity of carbonaceous microfossils can be relatively difficult because of their small size and simple shape, and to the degradation of the associated organic molecules with time. For Precambrian fossils, it generally requires the use of several techniques to study the shape and the composition of the structure itself, as well as its mineral environment. The ability to identify both organic matter and minerals using Raman spectroscopy makes it a key technique in the field of micropaleontology. Raman instruments are also being developed for the upcoming missions to Mars, ExoMars and Mars 2020, both dedicated to the search for past or present traces of life. However, demonstrating the biotic origin of carbonaceous matter in geological materials using this technique is controversial. Here, we show that Raman mapping instead of single spot analysis can detect variations in the composition of carbonaceous matter associated with fossilized microbes in the 800-Ma-old microfossils from the Draken Formation, Svalbard. This discovery is of great interest because it permits assessment of the biotic origin of a fossilized carbonaceous structure. Raman mapping could thus be of crucial importance in the near future for detecting potential fossilized microbial remains in Martian rocks.

Siliceous hot-spring deposits, or sinters, typically form in active, terrestrial (on land), volcanic terrains where magmatically heated waters circulating through the shallow crust emerge at the Earth’s surface as silica-charged geothermal fluids. Geyserites are sinters affiliated with the highest temperature ( 75–100 °C), natural geothermal fluid emissions, comprising localized, lithologically distinctive, hydrothermal silica precipitates that develop around geysers, spouters and spring-vents. They demarcate the position of hot-fluid upflow zones useful for geothermal energy and epithermal mineral prospecting. Near-vent areas also are “extreme environment” settings for the growth of microbial biofilms at near-boiling temperatures. Microbial biosignatures (e.g., characteristic silicified microbial textures, carbon isotopes, genetic material, lipid biomarkers) may be extracted from modern geyserite. However, because of strong taphonomic filtering and subsequent diagenesis, fossils in geyserite are very rare in the pre-Quaternary sinter record which, in and of itself, is patchy in time and space back to about 400 Ma. Only a few old examples are known, such as geyserite reported from the Devonian Drummond Basin (Australia), Devonian Rhynie cherts (Scotland), and a new example described herein from the spectacularly well-preserved, Late Jurassic (150 Ma), Yellowstone-style geothermal landscapes of Patagonia, Argentina. There, geyserite is associated with fossil vent-mounds and silicified hydrothermal breccias of the Claudia sinter, which is geologically related to the world-class Cerro Vanguardia gold/silver deposit of the Deseado Massif, a part of the Chon Aike siliceous large igneous province. Tubular, filament-like micro-inclusions from Claudia were studied using integrated petrographic and laser micro-Raman analysis, the results of which suggest a biological origin. The putative fossils are enclosed within nodular geyserite, a texture typical of subaerial near-vent conditions. Overall, this worldwide review of geyserite confirms its significance as a mineralizing geological archive reflecting the nature of Earth’s highest temperature, habitable terrestrial sedimentary environment. Hot-spring depositional settings also may serve as analogs for early Earth paleoenvironments because of their elevated temperature of formation, rapid mineralization by silica, and morphologically comparable carbonaceous material sourced from prokaryotes adapted to life at high temperatures.

The future ExoMars rover mission (ESA/Roscosmos), to be launched in 2018, will investigate the habitability of the Martian surface and near subsurface, and search for traces of past life in the form of textural biosignatures and organic molecules. In support of this mission, a selection of relevant Mars analogue materials has been characterised and stored in the International Space Analogue Rockstore (ISAR), hosted in Orléans, France. Two ISAR samples were analysed by prototypes of the ExoMars rover instruments used for petrographic study. The objective was to determine whether a full interpretation of the rocks could be achieved on the basis of the data obtained by the ExoMars visible-IR imager and spectrometer (MicrOmega), the close-up imager (CLUPI), the drill infrared spectrometer (Ma_Miss) and the Raman spectrometer (RLS), first separately then in their entirety. In order to not influence the initial instrumental interpretation, the samples were sent to the different teams without any additional information. This first step was called the “Blind Test” phase. The data obtained by the instruments were then complemented with photography of the relevant outcrops (as would be available during the ExoMars mission) before being presented to two geologists tasked with the interpretation. The context data and photography of the outcrops and of the samples were sufficient for the geologists to identify the rocks. This initial identification was crucial for the subsequent, iterative interpretation of the spectroscopic data. The data from the different spectrometers was, thus, cross-calibrated against the photographic interpretations and against each other. In this way, important mineralogical details, such as evidence of aqueous alteration of the rocks, provided relevant information concerning potential habitable conditions. The final conclusion from this test is that, when processed together, the ExoMars payload instruments produce complementary data allowing reliable interpretation of the geological context and potential for habitable environments. This background information is fundamental for the analysis and interpretation of organics in the processed Martian rocks.

Metazoans (multicellular animals) evolved during the Ediacaran Period as shown by the record of their imprints, carbonaceous compressions, trace fossils, and organic bodies and skeletal fossils. Initial evolutionary experiments produced unusual bodies that are poorly understood or conceived of as non-metazoan. It is accepted that sponges, ctenophorans, cnidarians, placozoans, and bilaterians were members of the Ediacaran fauna, many of which have uncertain affinities. The fossil Sabellidites cambriensis Yanishevsky, 1926, derived from the terminal Ediacaran strata, is the earliest known organically preserved animal that belonged to a newly evolving fauna, which replaced the Ediacara-type metazoans. Morphologically simple soft-bodied tubular fossils, such as S. cambriensis, and biomineralized, as contemporaneous Sinotubulites sp., are not easy to recognize phylogenetically because many unrelated organisms developed encasing tubes independently. Therefore, in addition to morphologic information, evidence derived from the microstructure of the organic wall and its biochemistry may be vital to resolving fossil origins and phylogenetic relationships. Here we present morphological, microstructural and biogeochemical studies on S. cambriensis using various microscopic and spectroscopic techniques, which provide new evidence that supports its siboglinid, annelidan affinity. The late Ediacaran age of Sabellidites fossil constrains the minimum age of siboglinids and the timing of the divergence of including them annelids by fossil record and this could be tested using molecular clock estimates. The fine microstructure of the organic tube in Sabellidites is multi-layered and has discrete layers composed of differently orientated and perfectly shaped fibers embedded in an amorphous matrix. The highly ordered and specific pattern of fiber alignment (i.e., the texture of organic matter) is similar to that of representatives of the family Siboglinidae. The biogeochemistry of the organic matter that comprised the tube, which was inferred from its properties, composition, and microstructure, is consistent with chitin and proteins as in siboglinids.

A suite of green and carbonaceous black chert interbedded with submarine volcanic rocks from several stratigraphic levels of the Onverwacht Group of the Barberton greenstone belt, from the top of the 3.48 Ga Komati Formation to the base of the 3.26 Ga Fig Tree Group have been investigated petrologically and geochemically. Provenance analysis was undertaken using immobile trace element ratios in comparison with potential source rocks from the Barberton granitoid-greenstone terrain. Raman spectroscopy was used to better characterise carbonaceous matter in different chert varieties. Green cherts consist predominantly of silicified mafic to ultramafic volcaniclastic material. Black cherts contain volcaniclastic and/or epiclastic material of ultramafic to felsic composition, admixed with carbonaceous matter. In several cases, the clastic sediment is compositionally distinct to the volcanic substrate, suggesting that it represents volcanic material of relatively distal sources. Soon after settling out of the water column, lithification due to silicification took place. It gave rise to excellent textural preservation, but strongly modified the mineralogical and chemical composition of the seafloor sediments. Zircon, Cr-spinel and rare phosphate minerals are the only primary minerals remaining, while clastic grains have been largely replaced by a fine intergrowth of microquartz, Ti-oxide and K-bearing mica. Chemical changes are similar to those reported previously for silicification of volcanic rocks from the barberton belt and are consistent with low-temperature hydrothermal processes common to the Palaeoarchaean seafloor environment.

The Raman Laser Spectrometer (RLS) is part of the payload of the 2018 ExoMars rover. The Sample Preparation and Distribution System (SPDS) of the rover will crush samples acquired from down to two meters depth under the Martian surface, and provide them to the RLS instrument in the form of flattened powdered samples. The RLS instrument will acquire a minimum of 20 points on the flattened surface of the samples. To be able to obtain the maximum scientific return from the instrument once on Mars, a simulator of the SPDS system has been built to perform a series of experiments in a representative scenario. The crushing process implies the loss of rock structure and texture and, hence, the geological context of the samples. However, qualitative analysis with the RLS simulator on powdered natural samples and rocks showed that the RLS is capable of detecting carbonaceous material occurring in trace amounts in one of the rock samples (a silicified volcanic sand), more easily than with the same analysis on bulk. Furthermore, it is shown that minor phases in carbonate cements that cannot be detected by Raman in the bulk sample can be detected in the powder, thus allowing the identification of all the carbonate phases present in the cement crust.

In order to quantify the detection threshold of the instrument, further analysis on controlled samples were performed. The results with the RLS SPDS simulator showed that the instrument can reach detection thresholds down to 1 % on powdered samples. Furthermore, analysis of controlled mixtures showed that performing a very simple intensity-based statistical analysis of the spectra can provide semi-quantification of the abundance of the mineral species with quite linear calibration curves.

Abstract Extraterrestrial habitability is a complex notion. We briefly review what is known about the origin of life on Earth, that is, life based on carbon chemistry and water. We then discuss habitable conditions (past and present) for established life and for the survival of microorganisms. Based on these elements, we propose to use the term habitable only for conditions necessary for the origin of life, the proliferation of life, and the survival of life. Not covered by this term would be conditions necessary for prebiotic chemistry and conditions that would allow the recognition of extinct or hibernating life. Finally, we apply this concept to the potential emergence of life on Mars where suitable conditions for life to start, proliferate, and survive have been heterogeneous throughout its history. These considerations have a profound impact on the nature and distribution of eventual traces of martian life, or any precursor, and must therefore inform our search-for-life strategies.

Instruments for surface missions to extraterrestrial bodies should be cross-calibrated using a common suite of relevant materials. Such work is necessary to improve instrument performance and aids in the interpretation of in-situ measurements. At the CNRS campus in Orléans, the Observatoire des Sciences de l’Univers en région Centre (OSUC) has created a collection of well-characterised rocks and minerals for testing and calibrating instruments to be flown in space missions. The characteristics of the analogue materials are documented in an accompanying online database. In view of the recent and upcoming rover missions to Mars (NASA’s 2011 Mars Science Laboratory (MSL) and ESA/Roscosmos’ 2018 ExoMars), we are concentrating initially on materials of direct relevance to the red planet. The initial collection consists of 15 well-studied rock and mineral samples, including a variety of basalts (ultramafic, weathered, silicified, primitive), sediments (volcanic sands, chert, and a banded iron formation –BIF-), and the phyllosilicate nontronite (a clay). All the samples were characterised petrographically, petrologically, and geochemically using the types of analyses likely to be performed during in-situ missions, in particular ExoMars : hand specimen description ; optical microscopy ; mineralogical analysis by XRD, Raman and IR spectrometry ; iron phase analysis by Mössbauer spectroscopy (MBS), elemental analysis by Energy-Dispersive X-ray spectroscopy (EDX), microprobe, Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) and Mass Spectrometry (ICP-MS) ; and reduced carbon analysis by Raman spectrometry.

Raman spectroscopy can be used for analysing both mineral and organic phases, thus allowing characterisation of the microbial-scale geological context as well as the search for possible traces of life. This method is therefore very useful for in situ planetary exploration missions. Compared with the myriad of sample preparation techniques available in terrestrial laboratories, the possibilities for sample preparation during in situ missions on other planetary bodies are extremely limited and are generally restricted to abrasion of rock surfaces or crushing of the target samples. Whereas certain techniques need samples to be prepared in powder form, such as X-ray diffraction, this kind of preparation is not particularly suitable for optical microscopy and/or Raman spectroscopy. In this contribution, we examine the effects of powdering rock and mineral samples on optical observations and Raman analyses. We used a commercial Raman spectrometer, as well as a Raman laser spectrometer that simulates the instrument being developed for the future ExoMars 2018 mission. The commercial Raman spectrometer documents significant modifications to the spectra of the powdered samples, including broadening of the peaks and shifts in their position, as well as the appearance of new peaks. These effects are caused by localised heating of the sample under the laser beam and amplification of nominal surface effects due to the increase in surface area in finer grain sizes. However, most changes observed in the Raman spectra using the Raman laser spectrometer system are negligible because the relatively large (50 µm diameter) laser spot size produces lower irradiance. Furthermore, minor phases were more easily detectable in the powdered samples. Most importantly, however, this sample preparation method results in the loss of the textural features and context, making identification of potential fossilized microbial remains more problematic.

Near-infrared emitting, magnetic particles for combined optical and MR detection based on liposomes or artificial lipoproteins are presented. They provide a novel strategy for the luminescence sensitization of lanthanide cations (Yb3+, Nd3+) without covalent bonds between the chromophore and the lanthanide, and provide an unambiguous tool for monitoring the integrity of the liponanoparticles, via emission in the NIR region.

Opaline silica was detected, with Raman spectroscopy, in carbonaceous microfossils (especially Myxococcoides) in silicified filamentous microbial mats within dolomitized conglomerates of the Draken Formation (- 800 to - 700 Ma). High-resolution electron microscopy (HRTEM) and microprobe analyses were used to confirm the nature of this phase in the quartz matrix of the microbial mats. The silica likely precipitated in a icrocrystalline form onto the organic macromolecules around, and within, the degrading microorganisms and preserved them by inhibiting the natural phase change to quartz. The Raman signal of opaline silica associated with carbonaceous matter and other iosignatures could be a potential indicator of biogenicity. This kind of association could be very useful during the future ExoMars mission (ESA/Roscosmos, 2018) that will earch for traces of past life on Mars.

Ironstones and iron-rich limestones regularly occur as components of time-specific intervals of the Palaeozoic as well as in younger times (Brett et al., this issue). Silurian sediments deposited at high latitudes along the peri-Gondwana border are characterized by black and white limestone and graptolitic shale sequences. Those in the Carnic Alps (southern Austria) additionally contain colourful pink to red limestones and ironstones. Laminated structures such as the (ferruginous)-coatings around skeletal fragments (mostly trilobites and some cephalopods and echinoderms) and stromatolitic features along discontinuity surfaces display dark red, green, white and brownish colours due to the presence of goethite,magnetite, hematite, chamosite, calcite and subordinate apatite.
Confocal laser Raman microscopy and complementary microscopic analysis of these ferruginous laminated structures document the presence of carbonaceousmatter associatedwith fossilizedmicrobial structures in the form of stromatolites, filaments and coccoids, suggesting a microbial role in the colouring of the Silurian world of the Carnic Alps. Iron concentrations up to 30× that of matrix and surrounding non-ferruginous rocks suggest blooms of iron microbe activity in response to the time-specific occurrence of chemically charged sea water during global biotic events.

Analyses by the Mars Exploration Rover (MER), Spirit, of Martian basalts from Gusev crater show that they are chemically very different from terrestrial basalts, being characterized in particular by high Mg- and Fe-contents. To provide suitable analog basalts for the International Space Analogue Rockstore (ISAR), a collection of analog rocks and minerals for preparing in situ space missions, especially, the upcoming Mars mission MSL- 2011 and the future international Mars-2018 mission, it is necessary to synthesize Martian basalts. The aim of this study was therefore to synthesize Martian basalt analogs to the Gusev crater basalts, based on the geochemical data from the MER rover Spirit. We present the results of two experiments, one producing a quench-cooled basalt (<1 h) and one producing a more slowly cooled basalt (1 day). Pyroxene and olivine textures produced in the more slowly cooled basalt were surprisingly similar to spinifex textures in komatiites, a volcanic rock type very common on the early Earth. These kinds of ultramafic rocks and their associated alteration products may have important astrobiological implications when associated with aqueous environments. Such rocks could provide habitats for chemolithotrophic microorganisms, while the glass and phyllosilicate derivatives can fix organic compounds.

Tailings and slag residues from the most important antimony mine of the French Massif Central were analysed for their mineralogical and chemical contents by conventional X-ray powder diffraction and synchrotron-based X-ray microdiffraction (μ-XRD). Results show that ∼2000 metric tons of Sb are still present at the abandoned mining site. Mean concentrations of Sb in slags and tailings are 1700 and 5000 mg kg−1, respectively. In addition, smaller quantities of As were also measured (∼800 mg kg−1 in tailings). Toxicity tests of As and Sb indicate that the growth of bacteria is severely affected at these concentrations. In particular, Sb was observed to cause negative effects for several types of bacteria. Almost all primary minerals carrying trivalent Sb disappeared during weathering at the expense of phases in which Sb5+ is the most abundant form. Instead of sulphides, Sb-bearing Fe hydroxides (goethite and lepidocrocite) are now present in the residues together with Sb-bearing jarosite and Sb(-Fe) oxides and hydroxides such as tripuhyite, senarmontite, romeite, cervantite, and valentinite. Water analyses of the main local stream indicate little remobilization of Sb downstream the site and despite the acidic pH of the surface tailings, pH values show neutral or near-neutral values on all locations of the site.

Timing the appearance of photosynthetic microorganisms is crucial to understanding the evolution of life on Earth. The ability of the biosphere to use sunlight as a source of energy (photoautotrophy) would have been essential for increasing biomass and for increasing the biogeochemical capacity of all prokaryotes across the range of redox reactions that support life. Typical proxies for photosynthesis in the rock record include features, such as a mat-like, laminated morphology (stratiform, domical, conical) often associated with bulk geochemical signatures, such as calcification, and a fractionated carbon isotope signature. However, to date, in situ, calcification related to photosynthesis has not been demonstrated in the oldest known microbial mats. We here use in situ nanometre-scale techniques to investigate the structural and compositional architecture in a 3.3 billion-year (Ga) old microbial biofilm from the Barberton greenstone belt, thus documenting in situ calcification that was most likely related to anoxygenic photosynthesis. The Josefsdal Chert Microbial Biofilm (JCMB) formed in a littoral (photic) environment. It is characterised by a distinct vertical structural and compositional organisation. The lower part is calcified in situ by aragonite, progressing upwards into uncalcified kerogen characterised by up to 1% sulphur, followed by an upper layer that contains intact filaments at the surface. Crystallites of pseudomorphed pyrite are also associated with the biofilm suggesting calcification related to the activity of heterotrophic sulphur reducing bacteria. In this anoxygenic, nutrient-limited environment, the carbon required by the sulphur reducing bacteria could only have been produced by photoautotrophy. We conclude that the Josfsdal Chert Microbial Biofilm was formed by a consortium of anoxygenic microorganisms, including photosynthesisers and sulphur reducing bacteria.

The chilled rinds of pillow basalt from the Ampere-Coral Patch Seamounts in the eastern North Atlantic were studied as a potential habitat of microbial life. A variety of putative biogenic structures, which include filamentous and spherical microfossil-like structures, were detected in K-phillipsite-filled amygdules within the chilled rinds. The filamentous structures (similar to 2.5 mu m in diameter) occur as K-phillipsite tubules surrounded by an Fe-oxyhydroxide (lepidocrocite) rich membranous structure, whereas the spherical structures (from 4 to 2 mu m in diameter) are associated with Ti oxide (anatase) and carbonaceous matter. Several lines of evidence indicate that the microfossil-like structures in the pillow basalt are the fossilized remains of microorganisms. Possible biosignatures include the carbonaceous nature of the spherical structures, their size distributions and morphology, the presence and distribution of native fluorescence, mineralogical and chemical composition, and environmental context. When taken together, the suite of possible biosignatures supports the hypothesis that the fossil-like structures are of biological origin. The vesicular microhabitat of the rock matrix is likely to have hosted a cryptoendolithic microbial community. This study documents a variety of evidence for past microbial life in a hitherto poorly investigated and underestimated microenvironment, as represented by the amygdules in the chilled pillow basalt rinds. This kind of endolithic volcanic habitat would have been common on the early rocky planets in our Solar System, such as Earth and Mars. This study provides a framework for evaluating traces of past life in vesicular pillow basalts, regardless of whether they occur on early Earth or Mars.

Nickel thin films deposited by sputtering methods on lithium fluoride single crystals have been strained to study the effect of the dislocation emergence on the mechanical behavior of coated materials. In these experiments, the samples had been investigated by in situ atomic force microscopy. It is shown that the buckling phenomenon preferentially occurs on the steps structures created by the dislocations coming from the substrate. In the frame of the Föppl-von Karman theory of thin plates, shape equations for the film on the area of emergence have been determined for the different levels of strain. These shape equations are compared to the experimental observations and it is concluded that the model needs to be modified to take into account the substrates effects.

Nickel thin films of various thicknesses deposited on lithium fluoride single crystals have been strained to study the effect of steps on the buckling phenomenon. In these experiments, the nanometer-scale steps result from the emergence process of dislocations coming from the substrate and piling-up at the film/substrate interface. The coated specimens were investigated by in situ atomic force microscopy. It is shown that the buckling phenomenon preferentially occurs just above the step structures in compression tests, while no buckling takes place in tension. From elastic energy consideration, it is concluded that the main effect of substrate plasticity is to strongly modify the critical stress for buckling, with no significant result on the adhesion properties. (c) 2007 Elsevier B.V. All rights reserved.

We report experimental atomic force microscopy observations and analytical modeling of buckling structures of thin films deposited on single crystal substrates. The formation of straight-sided blisters just above the step structures resulting from the dislocations emergence has been observed and explained in the framework of the Foppl-von Karman theory of thin plates. A critical step height above which the buckling may occur has been determined and the asymmetry of the resulting blisters has been explained. Finally, the new buckling criterion has been compared with the classical one in the plane case and allows us to explain the blisters localization on step structures.

The postbuckling transition from an initially straight-sided wrinkle to a distribution of bubbles has been investigated by means of finite element simulations in the case of a thin film relying on a rigid substrate. The calculations show that a snapthrough occurs when the buckling wavelength exceeds a critical value. Experimental atomic force microscopy observations of this transition have been reported and found to be in good agreement with the calculations. (c) 2005 American Institute of Physics.

The post-critical regime of straight-sided wrinkles on compliant substrates of polycarbonate has been observed by atomic force microscope and investigated by means of finite element simulations. The effect of coupling between the film and its substrate has revealed a global buckling phenomenon, characterized by critical loads lower than those found in the case of a rigid substrate. Characteristic shapes of the buckled structure have been also found to spread over a region wider than the delaminated zone itself. A law relating the film deflexion to the stress has finally been established for any film/substrate system. (C) 2004 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.