Immunological detection of small organic molecules in the presence of perchlorates: relevance to the life marker chip and life detection on Mars

The proposed ExoMars mission, due to launch in 2018, aims to look for evidence of extant and extinct life in martian rocks and regolith. Previous attempts to detect organic molecules of biological or abiotic origin on Mars have been unsuccessful, which may be attributable to destruction of these molecules by perchlorate salts during pyrolysis sample extraction techniques. Organic molecules can also be extracted and measured with solvent-based systems. The ExoMars payload includes the Life Marker Chip (LMC) instrument, capable of detecting biomarker molecules of extant and extinct Earth-like life in liquid extracts of martian samples with an antibody microarray assay. The aim of the work reported here was to investigate whether the presence of perchlorate salts, at levels similar to those at the NASA Phoenix landing site, would compromise the LMC extraction and detection method. To test this, we implemented an LMC-representative sample extraction process with an LMC-representative antibody assay and used these to extract and analyze a model sample that consisted of a Mars analog sample matrix (JSC Mars-1) spiked with a representative organic molecular target (pyrene, an example of abiotic meteoritic infall targets) in the presence of perchlorate salts. We found no significant change in immunoassay function when using pyrene standards with added perchlorate salts. When model samples spiked with perchlorate salts were subjected to an LMC-representative liquid extraction, immunoassays functioned in a liquid extract and detected extracted pyrene. For the same model sample matrix without perchlorate salts, we observed anomalous assay signals that coincided with yellow coloration of the extracts. This unexpected observation is being studied further. This initial study indicates that the presence of perchlorate salts, at levels similar to those detected at the NASA Phoenix landing site, is unlikely to prevent the LMC from extracting and detecting organic molecules from martian samples.     

The physics,biology, and environmental ethics of making mars habitable

The considerable evidence that Mars once had a wetter, more clement, environment motivates the search for past or present life on that planet. This evidence also suggests the possibility of restoring habitable conditions on Mars. While the total amounts of the key molecules--carbon dioxide, water, and nitrogen--needed for creating a biosphere on Mars are unknown, estimates suggest that there may be enough in the subsurface. Super greenhouse gases, in particular, perfluorocarbons, are currently the most effective and practical way to warm Mars and thicken its atmosphere so that liquid water is stable on the surface. This process could take approximately 100 years. If enough carbon dioxide is frozen in the South Polar Cap and absorbed in the regolith, the resulting thick and warm carbon dioxide atmosphere could support many types of microorganisms, plants, and invertebrates. If a planet-wide martian biosphere converted carbon dioxide into oxygen with an average efficiency equal to that for Earth's biosphere, it would take > 100,000 years to create Earth-like oxygen levels. Ethical issues associated with bringing life to Mars center on the possibility of indigenous martian life and the relative value of a planet with or without a global biosphere.     

Using electrical impedance spectroscopy to detect water in planetary regoliths

We present data in examination of the utility of electrical impedance spectroscopy measurements for in situ surveys to determine the water content, distribution, and phase in unconsolidated planetary regolith. We conducted calibration experiments under conditions relevant to Mars: the concentration of electrolytes in solution was varied up to 1 M to simulate the effects of unsaturated dissolved minerals and brines. We also varied the water content of heterogeneous water/sand mixtures, made with these electrolytic solutions from 0.01 wt% to 10 wt%. Tests were performed at temperatures from +25 degrees C to -65 degrees C. Conductivity and dielectric permittivity calculated from the impedance measurements indicate an expected dependence on electrolyte concentration and relative independence from electrolyte type for both liquid water and water ice. Conductivity and calculated dielectric relaxation times for these aqueous solutions agree with existing data in the literature. The relative permittivity for heterogeneous water/sand mixtures is dominated by polarization effects for the electrode configuration used. However, the characteristic orientational relaxation of ice is still visible. The conductivity retains the strong dependence on electrolyte concentration, and the permittivity is still not affected by electrolyte type. A "universal" curve between conductivity and water content establishes detectability limits of <0.01 wt% and approximately 0.3 wt% for water/sand mixtures containing liquid water and ice, respectively.     

Survival of Bacillus subtilis endospores on ultraviolet-irradiated rover wheels and Mars regolith under simulated Martian conditions

Endospores of Bacillus subtilis HA101 were applied to a simulated Mars Exploration Rover (MER) wheel and exposed to Mars-normal UV irradiation for 1, 3, or 6 h. The experiment was designed to simulate a contaminated rover wheel sitting on its landing platform before rolling off onto the martian terrain, as was encountered during the Spirit and Opportunity missions. When exposed to 1 h of Mars UV, a reduction of 81% of viable endospores was observed compared to the non-UV irradiated controls. When exposed for 3 or 6 h, reductions of 94.6% and 96.6%, respectively, were observed compared to controls. In a second experiment, the contaminated rover wheel was rolled over a bed of heat-sterilized Mars analog soil; then the analog soil was exposed to full martian conditions of UV irradiation, low pressure (6.9 mbar), low temperature (-10°C), and an anaerobic CO(2) martian atmosphere for 24 h to determine whether endospores of B. subtilis on the contaminated rover wheel could be transferred to the surface of the analog soil and survive martian conditions. The experiment simulated conditions in which a rover wheel might come into contact with martian regolith immediately after landing, such as is designed for the upcoming Mars Science Laboratory (MSL) rover. The contaminated rover wheel transferred viable endospores of B. subtilis to the Mars analog soil, as demonstrated by 31.7% of samples showing positive growth. However, when contaminated soil samples were exposed to full martian conditions for 24 h, only 16.7% of samples exhibited positive growth-a 50% reduction in the number of soil samples positive for the transferred viable endospores.     

Subfreezing activity of microorganisms and the potential habitability of Mars' polar regions

The availability of water-ice at the surface in the Mars polar cap and within the top meter of the high-latitude regolith raises the question of whether liquid water can exist there under some circumstances and possibly support the existence of biota. We examine the minimum temperatures at which liquid water can exist at ice grain-dust grain and ice grain-ice grain contacts, the minimum subfreezing temperatures at which terrestrial organisms can grow or multiply, and the maximum temperatures that can occur in martian high-latitude and polar regions, to see if there is overlap. Liquid water can exist at grain contacts above about -20 degrees C. Measurements of growth in organisms isolated from Siberian permafrost indicate growth at -10 degrees C and metabolism at -20 degrees C. Mars polar and high-latitude temperatures rise above -20 degrees C at obliquities greater than ~40 degrees, and under some conditions rise above 0 degrees C. Thus, the environment in the Mars polar regions has overlapped habitable conditions within relatively recent epochs, and Mars appears to be on the edge of being habitable at present. The easy accessibility of the polar surface layer relative to the deep subsurface make these viable locations to search for evidence of life.     

Searching for life on Mars: selection of molecular targets for ESA's aurora ExoMars mission

The European Space Agency's ExoMars mission will seek evidence of organic compounds of biological and non-biological origin at the martian surface. One of the instruments in the Pasteur payload may be a Life Marker Chip that utilizes an immunoassay approach to detect specific organic molecules or classes of molecules. Therefore, it is necessary to define and prioritize specific molecular targets for antibody development. Target compounds have been selected to represent meteoritic input, fossil organic matter, extant (living, recently dead) organic matter, and contamination. Once organic molecules are detected on Mars, further information is likely to derive from the detailed distribution of compounds rather than from single molecular identification. This will include concentration gradients beneath the surface and gradients from generic to specific compounds. The choice of biomarkers is informed by terrestrial biology but is wide ranging, and nonterrestrial biology may be evident from unexpected molecular distributions. One of the most important requirements is to sample where irradiation and oxidation are minimized, either by drilling or by using naturally excavated exposures. Analyzing regolith samples will allow for the search of both extant and fossil biomarkers, but sequential extraction would be required to optimize the analysis of each of these in turn.     

The Urey instrument: an advanced in situ organic and oxidant detector for Mars exploration

The Urey organic and oxidant detector consists of a suite of instruments designed to search for several classes of organic molecules in the martian regolith and ascertain whether these compounds were produced by biotic or abiotic processes using chirality measurements. These experiments will also determine the chemical stability of organic molecules within the host regolith based on the presence and chemical reactivity of surface and atmospheric oxidants. Urey has been selected for the Pasteur payload on the European Space Agency's (ESA's) upcoming 2013 ExoMars rover mission. The diverse and effective capabilities of Urey make it an integral part of the payload and will help to achieve a large portion of the mission's primary scientific objective: "to search for signs of past and present life on Mars." This instrument is named in honor of Harold Urey for his seminal contributions to the fields of cosmochemistry and the origin of life.     

Magnetism, iron minerals, and life on Mars

A short critical review is provided on two questions linking magnetism and possible early life on Mars: (1) Did Mars have an Earth-like internal magnetic field, and, if so, during which period and was it a requisite for life? (2) Is there a connection between iron minerals in the martian regolith and life? We also discuss the possible astrobiological implications of magnetic measurements at the surface of Mars using two proposed instruments. A magnetic remanence device based on magnetic field measurements can be used to identify Noachian age rocks and lightning impacts. A contact magnetic susceptibility probe can be used to investigate weathering rinds on martian rocks and identify meteorites among the small regolith rocks. Both materials are considered possible specific niches for microorganisms and, thus, potential astrobiological targets. Experimental results on analogues are presented to support the suitability of such in situ measurements.     

2-氯-4,6-二硝基-1,3-苯二酚萃取热力学研究

采用萃取平衡实验法,研究了用有机溶剂萃取水相中2-氯-4,6-二硝基-1,3-苯二酚的萃取热力学性质。考察了选用乙酸乙酯作萃取剂,苯、甲苯、环己烷和正庚烷作稀释剂时对萃取水相中2-氯-4,6-二硝基-1,3-苯二酚的影响。结果表明,萃合物的结构与稀释剂的种类有关,并且2-氯-4,6-二硝基-1,3-苯二酚在有机相中的分配系数随稀释剂介电常数的增大而增大。通过研究温度对萃取过程的影响,证实了乙酸乙酯从水相中萃取2-氯-4,6-二硝基-1,3-苯二酚的过程是一个放热过程,表观热效应ΔH=-10.088 kJ/mol。     

Bacterial growth at the high concentrations of magnesium sulfate found in martian soils

The martian surface environment exhibits extremes of salinity, temperature, desiccation, and radiation that would make it difficult for terrestrial microbes to survive. Recent evidence suggests that martian soils contain high concentrations of MgSO? minerals. Through warming of the soils, meltwater derived from subterranean ice-rich regolith may exist for an extended period of time and thus allow the propagation of terrestrial microbes and create significant bioburden at the near surface of Mars. The current report demonstrates that halotolerant bacteria from the Great Salt Plains (GSP) of Oklahoma are capable of growing at high concentrations of MgSO? in the form of 2 M solutions of epsomite. The epsotolerance of isolates in the GSP bacterial collection was determined, with 35% growing at 2 M MgSO?. There was a complex physiological response to mixtures of MgSO? and NaCl coupled with other environmental stressors. Growth also was measured at 1 M concentrations of other magnesium and sulfate salts. The complex responses may be partially explained by the pattern of chaotropicity observed for high-salt solutions as measured by agar gelation temperature. Select isolates could grow at the high salt concentrations and low temperatures found on Mars. Survival during repetitive freeze-thaw or drying-rewetting cycles was used as other measures of potential success on the martian surface. Our results indicate that terrestrial microbes might survive under the high-salt, low-temperature, anaerobic conditions on Mars and present significant potential for forward contamination. Stringent planetary protection requirements are needed for future life-detection missions to Mars.     

Survival of amino acids in micrometeorites during atmospheric entry

The delivery of amino acids by micrometeorites to the early Earth during the period of heavy bombardment could have been a significant source of the Earth's prebiotic amino acid inventory provided that these organic compounds survived atmospheric entry heating. To investigate the sublimation of amino acids from a micrometeorite analog at elevated temperature, grains from the CM-type carbonaceous chondrite Murchison were heated to 550 degrees C inside a glass sublimation apparatus (SA) under reduced pressure. The sublimed residue that had collected on the cold finger of the SA after heating was analyzed for amino acids by HPLC. We found that when the temperature of the meteorite reached approximately 150 degrees C, a large fraction of the amino acid glycine had vaporized from the meteorite, recondensed onto the end of the SA cold finger, and survived as the rest of the grains heated to 550 degrees C. alpha-Aminoisobutryic acid and isovaline, which are two of the most abundant non-protein amino acids in Murchison, did not sublime from the meteorite and were completely destroyed during the heating experiment. Our experimental results suggest that sublimation of glycine present in micrometeorite grains may provide a way for this amino acid to survive atmospheric entry heating at temperatures > 550 degrees C; all other amino acids apparently are destroyed.     

An inversion approach for lunar regolith layer thickness using optical albedo data and microwave emission simulation

An approach to inversion of the lunar regolith layer thickness by using multi-channel brightness temperature observation in passive microwave remote sensing is developed. To first make simulation of brightness temperature from the lunar layered media, the lunar regolith layer thickness (d) is proposed being constructed by available lunar DEM (digital elevation mapping) and on site measurements. The physical temperature distribution (T) over the lunar surface is also empirically assumed as a monotonic function of the latitude. Optical albedo of the lunar nearside from the telescopic observation is employed to construct the spatial distribution of the FeO+TiO₂ content (S) in the lunar regolith layer. A statistic relationship between the DEM and S of the lunar nearside is further extended to construction of S of the lunar farside. Thus, the dielectric permittivity (ε) of global lunar regolith layer can then be determined. Based on all these conditions (d,T,ε), brightness temperature of the lunar regolith layer in passive microwave remote sensing, which is planned for China's Chang-E lunar project, is numerically simulated by a parallel layering model using the strong fluctuation theory of random media.Then, taking these simulations with random noise as observations, an inversion method of the lunar regolith layer thickness is developed by using three- or two-channels brightness temperatures. When the S is low, and the four channels brightness temperatures in China's Chang-E project are well distinguishable, the regolith layer thickness and physical temperature of the underlying lunar rock media can be inverted by the three-channels approach. When the S becomes high that the brightness temperature at high frequency channels such as 19.35, 37 GHz are saturated, the regolith layer thickness is alternatively inverted only by the two-channels approach.Numerical simulation and inversion approach in this paper make an evaluation of the performance for lunar passive microwave remote sensing, and for future data calibration and validation.     

Brines in seepage channels as eluants for subsurface relict biomolecules on Mars?

Water, vital for life, not only maintains the integrity of structural and metabolic biomolecules, it also transports them in solution or colloidal suspension. Any flow of water through a dormant or fossilized microbial community elutes molecules that are potentially recognizable as biomarkers. We hypothesize that the surface seepage channels emanating from crater walls and cliffs in Mars Orbiter Camera images results from fluvial erosion of the regolith as low-temperature hypersaline brines. We propose that, if such flows passed through extensive subsurface catchments containing buried and fossilized remains of microbial communities from the wet Hesperian period of early Mars (approximately 3.5 Ga ago), they would have eluted and concentrated relict biomolecules and delivered them to the surface. Life-supporting low-temperature hypersaline brines in Antarctic desert habitats provide a terrestrial analog for such a scenario. As in the Antarctic, salts would likely have accumulated in water-filled depressions on Mars by seasonal influx and evaporation. Liquid water in the Antarctic cold desert analogs occurs at -80 degrees C in the interstices of shallow hypersaline soils and at -50 degrees C in salt-saturated ponds. Similarly, hypersaline brines on Mars could have freezing points depressed below -50 degrees C. The presence of hypersaline brines on Mars would have extended the amount of time during which life might have evolved. Phototrophic communities are especially important for the search for life because the distinctive structures and longevity of their pigments make excellent biomarkers. The surface seepage channels are therefore not only of geomorphological significance, but also provide potential repositories for biomolecules that could be accessed by landers.     

Martian CH(4): sources, flux, and detection

Recent observations have detected trace amounts of CH(4) heterogeneously distributed in the martian atmosphere, which indicated a subsurface CH(4) flux of ~2 x 10(5) to 2 x 10(9) cm(2) s(1). Four different origins for this CH(4) were considered: (1) volcanogenic; (2) sublimation of hydrate- rich ice; (3) diffusive transport through hydrate-saturated cryosphere; and (4) microbial CH(4) generation above the cryosphere. A diffusive flux model of the martian crust for He, H(2), and CH(4) was developed based upon measurements of deep fracture water samples from South Africa. This model distinguishes between abiogenic and microbial CH(4) sources based upon their isotopic composition, and couples microbial CH(4) production to H(2) generation by H(2)O radiolysis. For a He flux of approximately 10(5) cm(2) s(1) this model yields an abiogenic CH(4) flux and a microbial CH(4) flux of approximately 10(6) and approximately 10(9) cm(2) s(1), respectively. This flux will only reach the martian surface if CH(4) hydrate is saturated in the cryosphere; otherwise it will be captured within the cryosphere. The sublimation of a hydrate-rich cryosphere could generate the observed CH(4) flux, whereas microbial CH(4) production in a hypersaline environment above the hydrate stability zone only seems capable of supplying approximately 10(5) cm(2) s(1) of CH(4). The model predicts that He/H(2)/CH(4)/C(2)H(6) abundances and the C and H isotopic values of CH(4) and the C isotopic composition of C(2)H(6) could reveal the different sources. Cavity ring-down spectrometers represent the instrument type that would be most capable of performing the C and H measurements of CH(4) on near future rover missions and pinpointing the cause and source of the CH(4) emissions.     

Condensation reactions and formation of amides, esters, and nitriles under hydrothermal conditions

Hydrothermal pyrolysis experiments were performed to assess condensation (dehydration) reactions to amide, ester, and nitrile functionalities from lipid precursors. Beside product formation, organic compound alteration and stability were also evaluated. Mixtures of nonadecanoic acid, hexadecanedioic acid, or hexadecanamide with water, ammonium bicarbonate, and oxalic acid were heated at 300 degrees C for 72 h. In addition, mixtures of ammonium bicarbonate and oxalic acid solutions were used to test the abiotic formation of organic nitrogen compounds at the same temperature. The resulting products were condensation compounds such as amides, nitriles, and minor quantities of N-methylalkyl amides, alkanols, and esters. Mixtures of alkyl amide in water or oxalic acid yielded mainly hydrolysis and dehydration products, and with ammonium bicarbonate and oxalic acid the yield of condensation products was enhanced. The synthesis experiments with oxalic acid and ammonium bicarbonate solutions yielded homologous series of alkyl amides, alkyl amines, alkanes, and alkanoic acids, all with no carbon number predominances. These organic nitrogen compounds are stable and survive under the elevated temperatures of hydrothermal fluids.     

Oxidant enhancement in martian dust devils and storms: storm electric fields and electron dissociative attachment

Laboratory studies, numerical simulations, and desert field tests indicate that aeolian dust transport can generate atmospheric electricity via contact electrification or "triboelectricity." In convective structures such as dust devils and dust storms, grain stratification leads to macroscopic charge separations and gives rise to an overall electric dipole moment in the aeolian feature, similar in nature to the dipolar electric field generated in terrestrial thunderstorms. Previous numerical simulations indicate that these storm electric fields on Mars can approach the ambient breakdown field strength of approximately 25 kV/m. In terrestrial dust phenomena, potentials ranging from approximately 20 to 160 kV/m have been directly measured. The large electrostatic fields predicted in martian dust devils and storms can energize electrons in the low pressure martian atmosphere to values exceeding the electron dissociative attachment energy of both CO2 and H2O, which results in the formation of the new chemical products CO/O- and OH/H-, respectively. Using a collisional plasma physics model, we present calculations of the CO/O- and OH/H- reaction and production rates. We demonstrate that these rates vary geometrically with the ambient electric field, with substantial production of dissociative products when fields approach the breakdown value of approximately 25 kV/m. The dissociation of H2O into OH/H- provides a key ingredient for the generation of oxidants; thus electrically charged dust may significantly impact the habitability of Mars.     

On laboratory simulation and the effect of small temperature oscillations about the freezing point and ice formation on the evaporation rate of water on Mars

We report measurements of the evaporation rate of water under Mars-like conditions (CO2 atmosphere at 7 mbar and approximately 0 degrees C) in which small temperature oscillations about the freezing point repeatedly formed and removed a thin layer of ice. We found that the average evaporation at 2.7 +/- 0.5 degrees C without an ice layer (corrected for the difference in gravity on Earth and on Mars) was 1.24 +/- 0.12 mm/h, while at -2.1 +/- 0.3 degrees C with an ice layer the average evaporation rate was 0.84 +/- 0.08 mm/h. These values are in good agreement with those calculated for the evaporation of liquid water and ice when it is assumed that evaporation only depends on diffusion and buoyancy. Our findings suggest that such differences in evaporation rates are entirely due to the temperature difference and that the ice layer has little effect on evaporation rate. We infer that the formation of thin layers of ice on pools of water on Mars does not significantly increase the stability of water on the surface of Mars.     

沥青基C/C复合材料的制备及氧化行为

采用超高压浸渍-炭化方法制备了沥青基C/C复合材料,并对其在1273～1773 K氧化后的力学性能进行了测试,借助扫描电镜研究了该材料的氧化形貌.实验结果表明,在恒温下,随着氧化时间的延长,试样弯曲强度和模量逐渐减小,弯曲强度和模量与时间呈线性关系;而在恒定时间条件下,随着氧化温度的升高,试样的弯曲强度和模量亦逐渐减小...     

Low-temperature ionizing radiation resistance of Deinococcus radiodurans and Antarctic Dry Valley bacteria

The high flux of cosmic rays onto the unshielded surface of Mars poses a significant hazard to the survival of martian microbial life. Here, we determined the survival responses of several bacterial strains to ionizing radiation exposure while frozen at a low temperature characteristic of the martian near-subsurface. Novel psychrotolerant bacterial strains were isolated from the Antarctic Dry Valleys, an environmental analogue of the martian surface, and identified by 16S rRNA gene phylogeny as representatives of Brevundimonas, Rhodococcus, and Pseudomonas genera. These isolates, in addition to the known radioresistant extremophile Deinococcus radiodurans, were exposed to gamma rays while frozen on dry ice (-79°C). We found D. radiodurans to exhibit far greater radiation resistance when irradiated at -79°C than was observed in similar studies performed at higher temperatures. This greater radiation resistance has important implications for the estimation of potential survival times of microorganisms near the martian surface. Furthermore, the most radiation resistant of these Dry Valley isolates, Brevundimonas sp. MV.7, was found to show 99% 16S rRNA gene similarity to contaminant bacteria discovered in clean rooms at both Kennedy and Johnson Space Centers and so is of prime concern to efforts in the planetary protection of Mars from our lander probes. Results from this experimental irradiation, combined with previous radiation modeling, indicate that Brevundimonas sp. MV.7 emplaced only 30?cm deep in martian dust could survive the cosmic radiation for up to 100,000 years before suffering 10? population reduction.     

Oxidant enhancement in martian dust devils and storms: implications for life and habitability

We investigate a new mechanism for producing oxidants, especially hydrogen peroxide (H2O2), on Mars. Large-scale electrostatic fields generated by charged sand and dust in the martian dust devils and storms, as well as during normal saltation, can induce chemical changes near and above the surface of Mars. The most dramatic effect is found in the production of H2O2 whose atmospheric abundance in the "vapor" phase can exceed 200 times that produced by photochemistry alone. With large electric fields, H2O2 abundance gets large enough for condensation to occur, followed by precipitation out of the atmosphere. Large quantities of H2O2 would then be adsorbed into the regolith, either as solid H2O2 "dust" or as re-evaporated vapor if the solid does not survive as it diffuses from its production region close to the surface. We suggest that this H2O2, or another superoxide processed from it in the surface, may be responsible for scavenging organic material from Mars. The presence of H2O2 in the surface could also accelerate the loss of methane from the atmosphere, thus requiring a larger source for maintaining a steady-state abundance of methane on Mars. The surface oxidants, together with storm electric fields and the harmful ultraviolet radiation that readily passes through the thin martian atmosphere, are likely to render the surface of Mars inhospitable to life as we know it.