Limits of life and the habitability of Mars: The ESA space experiment BIOMEX on the ISS

BIOMEX (BIOlogy and Mars EXperiment) is an ESA/Roscosmos space exposure experiment housed within the exposure facility EXPOSE-R2 outside the Zvezda module on the International Space Station (ISS). The design of the multiuser facility supports—among others—the BIOMEX investigations into the stability and level of degradation of space-exposed biosignatures such as pigments, secondary metabolites, and cell surfaces in contact with a terrestrial and Mars analog mineral environment. In parallel, analysis on the viability of the investigated organisms has provided relevant data for evaluation of the habitability of Mars, for the limits of life, and for the likelihood of an interplanetary transfer of life (theory of lithopanspermia). In this project, lichens, archaea, bacteria, cyanobacteria, snow/permafrost algae, meristematic black fungi, and bryophytes from alpine and polar habitats were embedded, grown, and cultured on a mixture of martian and lunar regolith analogs or other terrestrial minerals. The organisms and regolith analogs and terrestrial mineral mixtures were then exposed to space and to simulated Mars-like conditions by way of the EXPOSE-R2 facility. In this special issue, we present the first set of data obtained in reference to our investigation into the habitability of Mars and limits of life. This project was initiated and implemented by the BIOMEX group, an international and interdisciplinary consortium of 30 institutes in 12 countries on 3 continents. Preflight tests for sample selection, results from ground-based simulation experiments, and the space experiments themselves are presented and include a complete overview of the scientific processes required for this space experiment and postflight analysis. The presented BIOMEX concept could be scaled up to future exposure experiments on the Moon and will serve as a pretest in low Earth orbit

Infrared nanoscopy and tomography of intracellular structures

Although techniques such as fluorescence-based super-resolution imaging or confocal microscopy simultaneously gather both morphological and chemical data, these techniques often rely on the use of localized and chemically specific markers. To eliminate this flaw, we have developed a method of examining cellular cross sections using the imaging power of scattering-type scanning near-field optical microscopy and Fourier-transform infrared spectroscopy at a spatial resolution far beyond the diffraction limit. Herewith, nanoscale surface and volumetric chemical imaging is performed using the intrinsic contrast generated by the characteristic absorption of mid-infrared radiation by the covalent bonds. We employ infrared nanoscopy to study the subcellular structures of eukaryotic (Chlamydomonas reinhardtii) and prokaryotic (Escherichia coli) species, revealing chemically distinct regions within each cell such as the microtubular structure of the flagellum. Serial 100 nm-thick cellular cross-sections were compiled into a tomogram yielding a three-dimensional infrared image of subcellular structure distribution at 20 nm resolution. The presented methodology is able to image biological samples complementing current fluorescence nanoscopy but at less interference due to the low energy of infrared radiation and the absence of labeling

SpectroCube: a European 6U nanosatellite spectroscopy platform for astrobiology and astrochemistry

SpectroCube is a CubeSat-based miniaturized in-situ space exposure platform for astrochemistry and astrobiology research. Within a 6 unit (6U, with 1U corresponding to 10 cm x 10 cm x 10 cm) nanosatellite structure, an infrared spectrometer is interfaced with a sample handling system to measure photochemical changes of organic molecules, representing important biomarkers for the detection of life in our solar system and beyond. Monitoring degradation profiles and photochemical reaction kinetics of such biomarkers allows to identify suitable search targets for current and future planetary exploration and life-detection missions. SpectroCube is designed to be launched into a highly elliptical orbit around Earth and therefore allows to expose samples to higher solar UV and energetic particle radiation levels than previous exposure platforms in low Earth orbit, as for example on the International Space Station. In-situ data will be telemetered back to Earth and compared with solar and planetary simulation experiments in ground-based laboratory. We here present the design of SpectroCube, the scientific payload and its subsystems. We demonstrate that with the miniaturisation potential of infrared spectroscopy it is possible to fit the entire optical setup plus a sample handling system for up to 60 individually contained and hermetically sealed samples within less than half of the volume of a 6U CubeSat structure. Therefore, the remaining volume can be entirely used for additional subsystems such as attitude control, propulsion, fuel, onboard computer and telemetry. The design of the scientific payload is based on a commercial off-the-shelf miniaturised Fourier-transform spectrometer consisting of an infrared light source, an interferometer and infrared detector units. The mechanical robustness and suitability of such a system for space applications was assessed. Shock and vibration testing of the mechanically most sensitive unit, the interferometer, was performed and revealed that with adequate damping the spectroscopic performance can be maintained. Additional measurements of test samples conducted with the selected commercial off-the-shelf spectrometer candidate showed that the spectroscopic range, resolution and sensitivity is capable to monitor in situ the photochemical kinetics of important classes of organic molecules and biomarkers for astrobiology and astrochemistry research

SPECTROModule: A modular in-situ spectroscopy platform for exobiology and space sciences

The evolution of the solar system and the origin of life remain some of the most intriguing questions for humankind. Addressing these questions experimentally is challenging due to the difficulty of mimicking environmental conditions representative for Early Earth and/or space conditions in general in ground-based laboratories. Performing experiments directly in space offers the great chance to overcome some of these obstacles and to possibly find answers to these questions. Exposure platforms in Low Earth Orbit (LEO) with the possibility for long-duration solar exposure are ideal for investigating the effects of solar and cosmic radiation on various biological and non-biological samples. Up to now, the Exobiology and space science research community has successfully made use of the International Space Station (ISS) via the EXPOSE facility to expose samples to the space environment with subsequent analyses after return to Earth. The emerging small and nanosatellite market represents another opportunity for astrobiology research as proven by the robotic O/OREOS mission, where samples were monitored in-situ, i.e. in Earth orbit. In this framework, the European Space Agency is developing a novel Exobiology facility outside the ISS. The new platform, which can host up to four different experiments, will combine the advantages of the ISS (long-term exposure, sample return capability) with near-real-time in-situ monitoring of the chemical/biological evolution in space. In particular, ultraviolet–visible (UV–Vis) and infrared (IR) spectroscopy were considered as key non-invasive methods to analyse the samples in situ. Changes in the absorption spectra of the samples developing over time will reveal the chemical consequences of exposure to solar radiation. Simultaneously, spectroscopy provides information on the growth rate or metabolic activities of biological cultures. The first quartet of experiments to be performed on-board consists of IceCold, OREOcube and Exocube (dual payload consisting of ExocubeChem and ExocubeBio). To prepare for the development of the Exobiology facility, ground units of the UV–Vis and IR spectrometers were studied, manufactured and tested as precursors of the flight units. The activity led to a modular in-situ spectroscopy platform able to perform different measurements (e.g. absorbance, optical density, fluorescence measurements) at the same time on different samples. We describe here the main features of the ground model platform, the verification steps, results and approach followed in the customization of commercial–off-the-shelf (COTS) modules to make them suitable for the space environment. The environmental tests included random and shock vibration, thermal vacuum cycles in the range −20 °C to +40 °C and irradiation of the components with a total dose of 1800 rad (18 Gy). The results of the test campaign consolidated the selection of the optical devices for the Exobiology Facility. The spectroscopic performance of the optical layout was tested and benchmarked in comparison with state-of-the-art laboratory equipment and calibration standards showing good correlation. This includes spectra of samples sets relevant for the flight experiments and a performance comparison between the SPECTROModule ground model and state-of-the-art laboratory spectrometers. Considering the large number of samples and different types of optical measurements planned on-board the ISS, the main outcome was the implementation of an LED-photodiode layout for the optical density and fluorescence measurements of IceCold (42 samples) and ExocubeBio (111 samples); while the UV–Vis spectrometer will be mainly focused on the change of the absorption spectra of the 48 samples of OREOcube.The ExocubeChem samples (in total 48) will be analysed by infrared spectroscopy. The ground platform supports the establishment of analogue research capabilities able to address the long-term objectives beyond the current application

Organism/Organic Exposure to Orbital Stresses (OOREOS) Satellite: Radiation Exposure in LEO and Supporting Laboratory Studies

We will present the results from the exposure of the metalloporphyrin iron tetraphenylporphyrin chloride (FeTPPCI), anthraufin (C(sub 14)H(sub 8)(O sub 4) (Anth) and Isoviolanthrene (C(sub 34H sub 18) (IVA) to the outher space environment, measured in situ aboard the Organism/Organic Exposure to Orbital Stresses nanosatellite. The compounds were exposed for a period of 17 months (3700 hours of direct solar exposure) including broad-spectrum solar radiation (approx. 122 nm to the near infrared). The organic films are enclosed in hermetically sealed sample cells that contain one of four astrobiologically relevant microenvironments. Transmission spectra (200-1000 nm) were recorded for each film, at first daily and subsequently every 15 days, along with a solar spectrum and the dark response of the detector array. In addition to analysis via UV-Vis spectroscopy, the laboratory controls were also monitored via infrared and far-UV spectroscopy. The results presented will include the finding that the FeTPPCI and IVA organic films in contact with a humid headspace gas (0.8-2.3%) exhibit faster degradation times, upon irradiation, in comparison with identical films under dry headspaces gases, whereas the Anth thin film exhibited a higher degree of photostability. In the companion laboratory experiments, simulated solar exposure of FeTPI films in contact with either Ar or CO(sub -2):O(sub -2):Ar (10:0.01:1000) headspace gas results in growth of a band in the films infrared spectra at 1961 cm(sup 1). Our assignment of this new spectral feature and the corresponding rational will be presented. The relevance of O/OREOS findings to planetary science, biomarker research, and the photostability of organic materials in astrobiologically relevant environments will also be discussed

Future space experiment platforms for astrobiology and astrochemistry research

Space experiments are a technically challenging but a scientifically important part of astrobiology and astrochemistry research. The International Space Station (ISS) is an excellent example of a highly successful and long-lasting research platform for experiments in space, that has provided a wealth of scientific data over the last two decades. However, future space platforms present new opportunities to conduct experiments with the potential to address key topics in astrobiology and astrochemistry. In this perspective, the European Space Agency (ESA) Topical Team Astrobiology and Astrochemistry (with feedback from the wider scientific community) identifies a number of key topics and summarizes the 2021 “ESA SciSpacE Science Community White Paper” for astrobiology and astrochemistry. We highlight recommendations for the development and implementation of future experiments, discuss types of in situ measurements, experimental parameters, exposure scenarios and orbits, and identify knowledge gaps and how to advance scientific utilization of future space-exposure platforms that are either currently under development or in an advanced planning stage. In addition to the ISS, these platforms include CubeSats and SmallSats, as well as larger platforms such as the Lunar Orbital Gateway. We also provide an outlook for in situ experiments on the Moon and Mars, and welcome new possibilities to support the search for exoplanets and potential biosignatures within and beyond our solar system

Proteomic and Metabolomic Characteristics of Extremophilic Fungi Under Simulated Mars Conditions

Filamentous fungi have been associated with extreme habitats, including nuclear power plant accident sites and the International Space Station (ISS). Due to their immense adaptation and phenotypic plasticity capacities, fungi may thrive in what seems like uninhabitable niches. This study is the first report of fungal survival after exposure of monolayers of conidia to simulated Mars conditions (SMC). Conidia of several Chernobyl nuclear accident-associated and ISS-isolated strains were tested for UV-C and SMC sensitivity, which resulted in strain-dependent survival. Strains surviving exposure to SMC for 30 min, ISSFT-021-30 and IMV 00236-30, were further characterized for proteomic, and metabolomic changes. Differential expression of proteins involved in ribosome biogenesis, translation, and carbohydrate metabolic processes was observed. No significant metabolome alterations were revealed. Lastly, ISSFT-021-30 conidia re-exposed to UV-C exhibited enhanced UV-C resistance when compared to the conidia of unexposed ISSFT-021

Left ventricular remodeling in swine after myocardial infarction: a transcriptional genomics approach

Despite the apparent appropriateness of left ventricular (LV) remodeling following myocardial infarction (MI), it poses an independent risk factor for development of heart failure. There is a paucity of studies into the molecular mechanisms of LV remodeling in large animal species. We took an unbiased molecular approach to identify candidate transcription factors (TFs) mediating the genetic reprogramming involved in post-MI LV remodeling in swine. Left ventricular tissue was collected from remote, non-infarcted myocardium, 3 weeks after MI-induction or sham-surgery. Microarray analysis identified 285 upregulated and 278 downregulated genes (FDR < 0.05). Of these differentially expressed genes, the promoter regions of the human homologs were searched for common TF binding sites (TFBS). Eighteen TFBS were overrepresented >two-fold (p < 0.01) in upregulated and 13 in downregulated genes. Left ventricular nuclear protein extracts were assayed for DNA-binding activity by protein/DNA array. Out of 345 DNA probes, 30 showed signal intensity changes >two-fold. Five TFs were identified in both TFBS and protein/DNA array analyses, which showed matching changes for COUP-TFII and glucocorticoid receptor (GR) only. Treatment of swine with the GR antagonist mifepristone after MI reduced the post-MI increase in LV mass, but LV dilation remained unaffected. Thus, using an unbiased approach to study post-MI LV remodeling in a physiologically relevant large animal model, we identified COUP-TFII and GR as potential key mediators of post-MI remodeling

BIOLEX – The Biology and Lunar experiment and the LOGOS Cubes

BIOLEX is a concept designed for in situ science on the Moon or in its orbit. As heritage of the polar and space experiment BIOMEX (Biology and Mars Experiment) on the ISS it is a more developed concept. Measurement operations on an exposure platform as well as within a micro-greenhouse device are part of this concept. The goal is to investigate the use of lunar resources as well as to analyse the stability of biomolecules as potential biosignatures serving as reference for future space exploration missions to Mars and the icy ocean moons in the outer solar system. Astrobiological exploration of the solar system is a priority research area such as emphasized by the European Astrobiology Roadmap (AstRoMap). It is focusing on several research topics, such as "Habitability" and on "Biomarkers for the detection of life". Therefore, "space platforms and laboratories", such as the EXPOSE setup installed outside the ISS, are essential to gain more knowledge on space- and planetary environments, which might be an essential basis for improvement of the robotic and human interplanetary exploration (Moon, Mars, Encedalus, Titan and Europa). In reference to these exposure platforms a new generation of hardware is needed to be installed in the lunar orbit or directly on the Moon. The BIOLEX is representing by its LOGOS (Lunar Organisms, Geo-microbiology and Organics Space Experiment) cubes such a concept combining the life detection topics with topics relevant to autonomous life supporting systems. A combination of a sample exposure device and a microhabitat for plants and microorganisms could address a tremendous number of questions from astrobiology and life sciences. The main scientific objectives for the use of BIOLEX-LOGOS cubes are: (i) in situ measurements by spectroscopy methods (such as Raman, IR, UV/VISspectroscopy) for analysis of biosignatures and their stability what is relevant for support of future life detection missions on Mars and the icy moons in the outer solar system); (ii) in situ measurements of environmental conditions (radiation, pressure/vacuum, temperature, pH, humidity) in micro-modules or compartments in reference to planned micro-habitat experiments placed on the Moon or incorporated on an exposure facility in orbit; (iii) in situ measurements of microorganisms’ activity in micro-modules / compartments in reference to planned microhabitat experiments placed on the moon or incorporated in the exposure facility in orbit. In reference to these scientific ideas the Moon is an excellent platform to operate different space experiments which will be of relevance for astrobiology, life sciences and human space missions. BIOLEX tries to fulfil a large number of scientific investigations in reference to these disciplines. The lunar environment is much harsher compared to Mars; and tests on biomolecules in this environment could provide information on their stability and therefore on the value to be used as reference for future space missions to Mars or the icy ocean moons in the outer solar system. Resources of the Moon such as the regolith or the freely available radiation on the surface could be tested by using them in a micro-greenhouse. Within this greenhouse different filters could test the optimal spectra range of the radiation