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1.  Mars for Earthlings: An Analog Approach to Mars in Undergraduate Education 
Astrobiology  2014;14(1):42-49.
Mars for Earthlings (MFE) is a terrestrial Earth analog pedagogical approach to teaching undergraduate geology, planetary science, and astrobiology. MFE utilizes Earth analogs to teach Mars planetary concepts, with a foundational backbone in Earth science principles. The field of planetary science is rapidly changing with new technologies and higher-resolution data sets. Thus, it is increasingly important to understand geological concepts and processes for interpreting Mars data. MFE curriculum is topically driven to facilitate easy integration of content into new or existing courses. The Earth-Mars systems approach explores planetary origins, Mars missions, rocks and minerals, active driving forces/tectonics, surface sculpting processes, astrobiology, future explorations, and hot topics in an inquiry-driven environment. Curriculum leverages heavily upon multimedia resources, software programs such as Google Mars and JMARS, as well as NASA mission data such as THEMIS, HiRISE, CRISM, and rover images.
Two years of MFE class evaluation data suggest that science literacy and general interest in Mars geology and astrobiology topics increased after participation in the MFE curriculum. Students also used newly developed skills to create a Mars mission team presentation. The MFE curriculum, learning modules, and resources are available online at Key Words: Mars—Geology—Planetary science—Astrobiology—NASA education. Astrobiology 14, 42–49.
PMCID: PMC3899945  PMID: 24359289
2.  Textural and Mineralogical Characteristics of Microbial Fossils Associated with Modern and Ancient Iron (Oxyhydr)Oxides: Terrestrial Analogue for Sediments in Gale Crater 
Astrobiology  2014;14(1):1-14.
Iron (oxyhydr)oxide microbial mats in modern to ∼100 ka tufa terraces are present in a cold spring system along Ten Mile Graben, southeastern Utah, USA. Mats exhibit morphological, chemical, and textural biosignatures and show diagenetic changes that occur over millennial scales. The Jurassic Brushy Basin Member of the Morrison Formation in the Four Corners region of the USA also exhibits comparable microbial fossils and iron (oxyhydr)oxide biosignatures in the lacustrine unit.
Both the modern spring system and Brushy Basin Member represent alkaline, saline, groundwater-fed systems and preserve diatoms and other similar algal forms with cellular elaboration. Two distinct suites of elements (1. C, Fe, As and 2. C, S, Se, P) are associated with microbial fossils in modern and ancient iron (oxyhydr)oxides and may be potential markers for biosignatures. The presence of ferrihydrite in ∼100 ka fossil microbial mats and Jurassic rocks suggests that this thermodynamically unstable mineral may also be a potential biomarker.
One of the most extensive sedimentary records on Mars is exposed in Gale Crater and consists of non-acidic clays and sulfates possibly of lacustrine origin. These terrestrial iron (oxyhydr)oxide examples are a valuable analogue because of similar iron- and clay-rich host rock compositions and will help (1) understand diagenetic processes in a non-acidic, saline lacustrine environment such as the sedimentary rocks in Gale Crater, (2) document specific biomediated textures, (3) demonstrate how biomediated textures might persist or respond to diagenesis over time, and (4) provide a ground truth library of textures to explore and compare in extraterrestrial iron (oxyhydr)oxides, where future explorations hope to detect past evidence of life. Key Words: Biogeochemistry—Mars—Biosignatures—Diagenesis—Iron oxides. Astrobiology 14, 1–14.
PMCID: PMC3901165  PMID: 24380534
3.  Spaceflight Induces Specific Alterations in the Proteomes of Arabidopsis 
Astrobiology  2015;15(1):32-56.
Life in spaceflight demonstrates remarkable acclimation processes within the specialized habitats of vehicles subjected to the myriad of unique environmental issues associated with orbital trajectories. To examine the response processes that occur in plants in space, leaves and roots from Arabidopsis (Arabidopsis thaliana) seedlings from three GFP reporter lines that were grown from seed for 12 days on the International Space Station and preserved on orbit in RNAlater were returned to Earth and analyzed by using iTRAQ broad-scale proteomics procedures. Using stringent criteria, we identified over 1500 proteins, which included 1167 leaf proteins and 1150 root proteins we were able to accurately quantify. Quantification revealed 256 leaf proteins and 358 root proteins that showed statistically significant differential abundance in the spaceflight samples compared to ground controls, with few proteins differentially regulated in common between leaves and roots. This indicates that there are measurable proteomics responses to spaceflight and that the responses are organ-specific. These proteomics data were compared with transcriptome data from similar spaceflight samples, showing that there is a positive but limited relationship between transcriptome and proteome regulation of the overall spaceflight responses of plants. These results are discussed in terms of emergence understanding of plant responses to spaceflight particularly with regard to cell wall remodeling, as well as in the context of deriving multiple omics data sets from a single on-orbit preservation and operations approach. Key Words: Space biology—Proteomics—Gene expression—ISS. Astrobiology 15, 32–56.
PMCID: PMC4290804  PMID: 25517942
4.  Explorer, Nobel Laureate, Astrobiologist: Things You Never Knew about Barry Blumberg 
Astrobiology  2015;15(1):1-14.
PMCID: PMC4291204  PMID: 25470005
5.  XUV-Exposed, Non-Hydrostatic Hydrogen-Rich Upper Atmospheres of Terrestrial Planets. Part I: Atmospheric Expansion and Thermal Escape 
Astrobiology  2013;13(11):1011-1029.
The recently discovered low-density “super-Earths” Kepler-11b, Kepler-11f, Kepler-11d, Kepler-11e, and planets such as GJ 1214b represent the most likely known planets that are surrounded by dense H/He envelopes or contain deep H2O oceans also surrounded by dense hydrogen envelopes. Although these super-Earths are orbiting relatively close to their host stars, they have not lost their captured nebula-based hydrogen-rich or degassed volatile-rich steam protoatmospheres. Thus, it is interesting to estimate the maximum possible amount of atmospheric hydrogen loss from a terrestrial planet orbiting within the habitable zone of late main sequence host stars. For studying the thermosphere structure and escape, we apply a 1-D hydrodynamic upper atmosphere model that solves the equations of mass, momentum, and energy conservation for a planet with the mass and size of Earth and for a super-Earth with a size of 2 REarth and a mass of 10 MEarth. We calculate volume heating rates by the stellar soft X-ray and extreme ultraviolet radiation (XUV) and expansion of the upper atmosphere, its temperature, density, and velocity structure and related thermal escape rates during the planet's lifetime. Moreover, we investigate under which conditions both planets enter the blow-off escape regime and may therefore experience loss rates that are close to the energy-limited escape. Finally, we discuss the results in the context of atmospheric evolution and implications for habitability of terrestrial planets in general. Key Words: Stellar activity—Low-mass stars—Early atmospheres—Earth-like exoplanets—Energetic neutral atoms—Ion escape—Habitability. Astrobiology 13, 1011–1029.
PMCID: PMC3865622  PMID: 24251443
6.  XUV-Exposed, Non-Hydrostatic Hydrogen-Rich Upper Atmospheres of Terrestrial Planets. Part II: Hydrogen Coronae and Ion Escape 
Astrobiology  2013;13(11):1030-1048.
We studied the interactions between the stellar wind plasma flow of a typical M star, such as GJ 436, and the hydrogen-rich upper atmosphere of an Earth-like planet and a “super-Earth” with a radius of 2 REarth and a mass of 10 MEarth, located within the habitable zone at ∼0.24 AU. We investigated the formation of extended atomic hydrogen coronae under the influences of the stellar XUV flux (soft X-rays and EUV), stellar wind density and velocity, shape of a planetary obstacle (e.g., magnetosphere, ionopause), and the loss of planetary pickup ions on the evolution of hydrogen-dominated upper atmospheres. Stellar XUV fluxes that are 1, 10, 50, and 100 times higher compared to that of the present-day Sun were considered, and the formation of high-energy neutral hydrogen clouds around the planets due to the charge-exchange reaction under various stellar conditions was modeled. Charge-exchange between stellar wind protons with planetary hydrogen atoms, and photoionization, lead to the production of initially cold ions of planetary origin. We found that the ion production rates for the studied planets can vary over a wide range, from ∼1.0×1025 s−1 to ∼5.3×1030 s−1, depending on the stellar wind conditions and the assumed XUV exposure of the upper atmosphere. Our findings indicate that most likely the majority of these planetary ions are picked up by the stellar wind and lost from the planet. Finally, we estimated the long-time nonthermal ion pickup escape for the studied planets and compared them with the thermal escape. According to our estimates, nonthermal escape of picked-up ionized hydrogen atoms over a planet's lifetime within the habitable zone of an M dwarf varies between ∼0.4 Earth ocean equivalent amounts of hydrogen (EOH) to <3 EOH and usually is several times smaller in comparison to the thermal atmospheric escape rates. Key Words: Stellar activity—Low-mass stars—Early atmospheres—Earth-like exoplanets—Energetic neutral atoms—Ion escape—Habitability. Astrobiology 13, 1030–1048.
PMCID: PMC3865724  PMID: 24283926
7.  Natural Fumarolic Alteration of Fluorapatite, Olivine, and Basaltic Glass, and Implications for Habitable Environments on Mars 
Astrobiology  2013;13(11):1049-1064.
Fumaroles represent a very important potential habitat on Mars because they contain water and nutrients. Global deposition of volcanic sulfate aerosols may also have been an important soil-forming process affecting large areas of Mars. Here we identify alteration from the Senator fumarole, northwest Nevada, USA, and in low-temperature environments near the fumarole to help interpret fumarolic and acid vapor alteration of rocks and soils on Mars. We analyzed soil samples and fluorapatite, olivine, and basaltic glass placed at and near the fumarole in in situ mineral alteration experiments designed to measure weathering under natural field conditions.
Using synchrotron X-ray diffraction, we clearly observe hydroxyl-carbonate-bearing fluorapatite as a fumarolic alteration product of the original material, fluorapatite. The composition of apatites as well as secondary phosphates has been previously used to infer magmatic conditions as well as fumarolic conditions on Mars. To our knowledge, the observations reported here represent the first documented instance of formation of hydroxyl-carbonate-bearing apatite from fluorapatite in a field experiment. Retreat of olivine surfaces, as well as abundant NH4-containing minerals, was also characteristic of fumarolic alteration. In contrast, alteration in the nearby low-temperature environment resulted in formation of large pits on olivine surfaces, which were clearly distinguishable from the fumarolic alteration. Raman signatures of some fumarolically impacted surfaces are consistent with detection of the biological molecules chlorophyll and scytenomin, potentially useful biosignatures. Observations of altered minerals on Mars may therefore help identify the environment of formation and understand the aqueous history and potential habitability of that planet. Key Words: Fumaroles—Mars—Olivine—Acidophile—Geothermal—Search for life (biosignatures)—Synchrotron X-ray diffraction. Astrobiology 13, 1049–1064.
PMCID: PMC3865726  PMID: 24283927
8.  Transfer of Impact Ejecta Material from the Surface of Mars to Phobos and Deimos 
Astrobiology  2013;13(10):963-980.
The Russian Phobos-Grunt spacecraft originally planned to return a 200 g sample of surface material from Phobos to Earth. Although it was anticipated that this material would mainly be from the body of Phobos, there is a possibility that such a sample may also contain material ejected from the surface of Mars by large impacts. An analysis of this possibility is completed by using current knowledge of aspects of impact cratering on the surface of Mars and the production of high-speed ejecta that might reach Phobos or Deimos. Key Words: Impact cratering—Ejecta transfer—Phobos. Astrobiology 13, 963–980.
PMCID: PMC3807532  PMID: 24131246
9.  Raman Hyperspectral Imaging of Microfossils: Potential Pitfalls 
Astrobiology  2013;13(10):920-931.
Initially, Raman spectroscopy was a specialized technique used by vibrational spectroscopists; however, due to rapid advancements in instrumentation and imaging techniques over the last few decades, Raman spectrometers are widely available at many institutions, allowing Raman spectroscopy to become a widespread analytical tool in mineralogy and other geological sciences. Hyperspectral imaging, in particular, has become popular due to the fact that Raman spectroscopy can quickly delineate crystallographic and compositional differences in 2-D and 3-D at the micron scale. Although this rapid growth of applications to the Earth sciences has provided great insight across the geological sciences, the ease of application as the instruments become increasingly automated combined with nonspecialists using this techique has resulted in the propagation of errors and misunderstandings throughout the field. For example, the literature now includes misassigned vibration modes, inappropriate spectral processing techniques, confocal depth of laser penetration incorrectly estimated into opaque crystalline solids, and a misconstrued understanding of the anisotropic nature of sp2 carbons. Key Words: Raman spectroscopy—Raman imaging—Confocal Raman spectroscopy—Disordered sp2 carbons—Hematite—Microfossils. Astrobiology 13, 920–931.
PMCID: PMC3807534  PMID: 24088070
10.  The Dependence of the Ice-Albedo Feedback on Atmospheric Properties 
Astrobiology  2013;13(10):899-909.
Ice-albedo feedback is a potentially important destabilizing effect for the climate of terrestrial planets. It is based on the positive feedback between decreasing surface temperatures, an increase of snow and ice cover, and an associated increase in planetary albedo, which then further decreases surface temperature. A recent study shows that for M stars, the strength of the ice-albedo feedback is reduced due to the strong spectral dependence of stellar radiation and snow/ice albedos; that is, M stars primarily emit in the near IR, where the snow and ice albedo is low, and less in the visible, where the snow/ice albedo is high.
This study investigates the influence of the atmosphere (in terms of surface pressure and atmospheric composition) on this feedback, since an atmosphere was neglected in previous studies. A plane-parallel radiative transfer model was used for the calculation of planetary albedos. We varied CO2 partial pressures as well as the H2O, CH4, and O3 content in the atmosphere for planets orbiting Sun-like and M type stars.
Results suggest that, for planets around M stars, the ice-albedo effect is significantly reduced, compared to planets around Sun-like stars. Including the effects of an atmosphere further suppresses the sensitivity to the ice-albedo effect. Atmospheric key properties such as surface pressure, but also the abundance of radiative trace gases, can considerably change the strength of the ice-albedo feedback. For dense CO2 atmospheres of the order of a few to tens of bar, atmospheric rather than surface properties begin to dominate the planetary radiation budget. At high CO2 pressures, the ice-albedo feedback is strongly reduced for planets around M stars. The presence of trace amounts of H2O and CH4 in the atmosphere also weakens the ice-albedo effect for both stellar types considered. For planets around Sun-like stars, O3 could also lead to a very strong decrease of the ice-albedo feedback at high CO2 pressures. Key Words: Atmospheric compositions—Extrasolar terrestrial planets—Snowball Earth—Planetary atmospheres—Radiative transfer. Astrobiology 13, 899–909.
PMCID: PMC3807702  PMID: 24111995
11.  Acidophilic Halophilic Microorganisms in Fluid Inclusions in Halite from Lake Magic, Western Australia 
Astrobiology  2013;13(9):850-860.
Lake Magic is one of the most extreme of hundreds of ephemeral acid-saline lakes in southern Western Australia. It has pH as low as 1.7, salinity as high as 32% total dissolved solids, temperatures ranging from 0°C to 50°C, and an unusually complex aqueous composition. Optical petrography, UV-vis petrography, and laser Raman spectrometry were used to detect microorganisms and organic compounds within primary fluid inclusions in modern bedded halite from Lake Magic. Rare prokaryotes appear as 1–3 μm, bright cocci that fluoresce green with UV-vis illumination. Dimpled, 5–7 μm yellow spherules that fluoresce blue with UV-vis illumination are interpreted as Dunaliella algae. Yellow-orange beta-carotene crystals, globules, and coatings are characterized by orange-red fluorescence and three distinct Raman peaks. Because acid saline lakes are good Mars analogues, the documentation of prokaryotes, eukaryotes, and organic compounds preserved in the halite here has implications for the search for life on Mars. Missions to Mars should incorporate such in situ optical and chemical examination of martian evaporites for possible microorganisms and/or organic compounds in fluid inclusions. Key Words: Acid—Extremophiles—Western Australia—Fluid inclusions—Lake Magic—Dunaliella. Astrobiology 13, 850–860.
PMCID: PMC3778947  PMID: 23971647
12.  The Effects of Modeled Microgravity on Growth Kinetics, Antibiotic Susceptibility, Cold Growth, and the Virulence Potential of a Yersinia pestis ymoA-Deficient Mutant and Its Isogenic Parental Strain 
Astrobiology  2013;13(9):821-832.
Previously, we reported that there was no enhancement in the virulence potential (as measured by cell culture infections) of the bacterial pathogen Yersinia pestis (YP) following modeled microgravity/clinorotation growth. We have now further characterized the effects of clinorotation (CR) on YP growth kinetics, antibiotic sensitivity, cold growth, and YP's virulence potential in a murine model of infection. Surprisingly, none of the aforementioned phenotypes were altered. To better understand why CR did not enhance YP's virulence potential as it did for other bacterial pathogens, a YP ΔymoA isogenic mutant in the KIM/D27 background strain that is unable to produce the histone-like YmoA protein and influences DNA topography was used in both cell culture and murine models of infection. YmoA represses type three secretion system (T3SS) virulence gene expression in the yersiniae. Similar to our CR-grown parental YP strain data, the CR-grown ΔymoA mutant induced reduced HeLa cell cytotoxicity with concomitantly decreased Yersinia outer protein E (YopE) and low calcium response V (LcrV) antigen production and secretion. Important, however, were our findings that, although no significant differences were observed in survival of mice infected intraperitoneally with either normal gravity (NG)- or CR-grown parental YP, the ΔymoA mutant induced significantly more mortality in infected mice than did the parental strain following CR growth. Taken together, our data demonstrate that CR did enhance the virulence potential of the YP ΔymoA mutant in a murine infection model (relative to the CR-grown parental strain), despite inducing less HeLa cell rounding in our cell culture infection assay due to reduced T3SS activity. Therefore, CR, which induces a unique type of bacterial stress, might be enhancing YP's virulence potential in vivo through a T3SS-independent mechanism when the histone-like YmoA protein is absent. Key Words: Type three secretion system (T3SS)—Low-shear modeled microgravity—High-aspect ratio vessel—Cell culture infection models—Mouse infection model. Astrobiology 13, 821–832.
PMCID: PMC3779001  PMID: 23988036
13.  Effects of Simulated Microgravity on Primary Human NK Cells 
Astrobiology  2013;13(8):703-714.
The deleterious effects of microgravity on lymphocytes have been demonstrated in previous studies. However, research on the effects of microgravity on human natural killer (NK) cells remains exceedingly limited. In this study, we demonstrated that NK cell cytotoxicity was significantly decreased under simulated microgravity (SMG) conditions (p<0.05). Several processes, including apoptosis, receptor expression, and cytokine secretion, were investigated in human NK cells under SMG. We observed decreased cytotoxicity, concurrent with increased apoptosis and necrosis, in NK cells after exposure to SMG (p<0.05). Additionally, interferon (IFN)-γ and perforin expression decreased significantly, and the expression of granzyme-B was only slightly reduced. Meanwhile, SMG selectively inhibited the expression of certain surface receptors on NK cells. Specifically, the expression of NKG2A and NKG2D were significantly downregulated under SMG, but the expression of NKp30 and NKp44 was not affected. We also found that interleukin (IL)–15 alone or in combination with IL-12 could counteract the inhibition of NK cell cytotoxicity under SMG. Our findings indicate that human NK cells were sensitive to SMG, as reflected by their decreased cytotoxicity. Factors such as increased early apoptosis and late apoptosis/necrosis and the decreased expression of INF-γ, cytolytic proteins, and cell surface receptors may be responsible for the loss of cytotoxicity in human NK cells under SMG. A combination of IL-12 and IL-15 may be useful as a therapeutic strategy for overcoming the effects of microgravity on human NK cells during long space missions. Key Words: Simulated microgravity (SMG)—Natural killer (NK) cells—Cytotoxicity. Astrobiology 13, 703–714.
PMCID: PMC3746215  PMID: 23919749
14.  Neurotoxic Potential of Lunar and Martian Dust: Influence on Em, Proton Gradient, Active Transport, and Binding of Glutamate in Rat Brain Nerve Terminals 
Astrobiology  2013;13(8):679-692.
The harmful effects of lunar dust (LD) on directly exposed tissues are documented in the literature, whereas researchers are only recently beginning to consider its effects on indirectly exposed tissues. During inhalation, nano-/microsized particles are efficiently deposited in nasal, tracheobronchial, and alveolar regions and transported to the central nervous system. The neurotoxic potential of LD and martian dust (MD) has not yet been assessed.
Glutamate is the main excitatory neurotransmitter involved in most aspects of normal brain function, whereas disturbances in glutamate homeostasis contribute to the pathogenesis of major neurological disorders. The research was focused on the analysis of the effects of LD/MD simulants (JSC-1a/JSC, derived from volcanic ash) on the key characteristics of glutamatergic neurotransmission. The average size of LD and MD particles (even minor fractions) before and after sonication was determined by dynamic light scattering. With the use of radiolabeled l-[14C]glutamate, it was shown that there is an increase in l-[14C]glutamate binding to isolated rat brain nerve terminals (synaptosomes) in low [Na+] media and at low temperature in the presence of LD. MD caused significantly lesser changes under the same conditions, whereas nanoparticles of magnetite had no effect at all. Fluorimetric experiments with potential-sensitive dye rhodamine 6G and pH-sensitive dye acridine orange showed that the potential of the plasma membrane of the nerve terminals and acidification of synaptic vesicles were not altered by LD/MD (and nanoparticles of magnetite).
Thus, the unique effect of LD to increase glutamate binding to the nerve terminals was shown. This can have deleterious effects on extracellular glutamate homeostasis in the central nervous system and cause alterations in the ambient level of glutamate, which is extremely important for proper synaptic transmission. During a long-term mission, a combination of constant irritation due to dust particles, inflammation, stress, low gravity and microgravity, radiation, UV, and so on may consequently change the effects of the dust and aggravate neurological consequences. Key Words: Lunar dust simulant—Martian dust simulant—Volcanic ash—Glutamate binding—Membrane potential—Synaptic vesicle acidification—Glutamatergic neurotransmission—Rat brain nerve terminals. Astrobiology 13, 679–692.
PMCID: PMC3746286  PMID: 23919751
15.  The Importance of Planetary Rotation Period for Ocean Heat Transport 
Astrobiology  2014;14(8):645-650.
The climate and, hence, potential habitability of a planet crucially depends on how its atmospheric and ocean circulation transports heat from warmer to cooler regions. However, previous studies of planetary climate have concentrated on modeling the dynamics of atmospheres, while dramatically simplifying the treatment of oceans, which neglects or misrepresents the effect of the ocean in the total heat transport. Even the majority of studies with a dynamic ocean have used a simple so-called aquaplanet that has no continental barriers, which is a configuration that dramatically changes the ocean dynamics. Here, the significance of the response of poleward ocean heat transport to planetary rotation period is shown with a simple meridional barrier—the simplest representation of any continental configuration. The poleward ocean heat transport increases significantly as the planetary rotation period is increased. The peak heat transport more than doubles when the rotation period is increased by a factor of ten. There are also significant changes to ocean temperature at depth, with implications for the carbon cycle. There is strong agreement between the model results and a scale analysis of the governing equations. This result highlights the importance of both planetary rotation period and the ocean circulation when considering planetary habitability. Key Words: Exoplanet—Oceans—Rotation—Climate—Habitability. Astrobiology 14, 645–650.
PMCID: PMC4126269  PMID: 25041658
16.  A Conspicuous Clay Ovoid in Nakhla: Evidence for Subsurface Hydrothermal Alteration on Mars with Implications for Astrobiology 
Astrobiology  2014;14(8):651-693.
A conspicuous biomorphic ovoid structure has been discovered in the Nakhla martian meteorite, made of nanocrystalline iron-rich saponitic clay and amorphous material. The ovoid is indigenous to Nakhla and occurs within a late-formed amorphous mesostasis region of rhyolitic composition that is interstitial to two clinopyroxene grains with Al-rich rims, and contains acicular apatite crystals, olivine, sulfides, Ti-rich magnetite, and a new mineral of the rhoenite group. To infer the origin of the ovoid, a large set of analytical tools was employed, including scanning electron microscopy and backscattered electron imaging, wavelength-dispersive X-ray analysis, X-ray mapping, Raman spectroscopy, time-of-flight secondary ion mass spectrometry analysis, high-resolution transmission electron microscope imaging, and atomic force microscope topographic mapping. The concentric wall of the ovoid surrounds an originally hollow volume and exhibits internal layering of contrasting nanotextures but uniform chemical composition, and likely inherited its overall shape from a preexisting vesicle in the mesostasis glass. A final fibrous layer of Fe-rich phases blankets the interior surfaces of the ovoid wall structure. There is evidence that the parent rock of Nakhla has undergone a shock event from a nearby bolide impact that melted the rims of pyroxene and the interstitial matter and initiated an igneous hydrothermal system of rapidly cooling fluids, which were progressively mixed with fluids from the melted permafrost. Sharp temperature gradients were responsible for the crystallization of Al-rich clinopyroxene rims, rhoenite, acicular apatites, and the quenching of the mesostasis glass and the vesicle. During the formation of the ovoid structure, episodic fluid infiltration events resulted in the precipitation of saponite rinds around the vesicle walls, altered pyrrhotite to marcasite, and then isolated the ovoid wall structure from the rest of the system by depositing a layer of iron oxides/hydroxides. Carbonates, halite, and sulfates were deposited last within interstitial spaces and along fractures. Among three plausible competing hypotheses here, this particular abiotic scenario is considered to be the most reasonable explanation for the formation of the ovoid structure in Nakhla, and although compelling evidence for a biotic origin is lacking, it is evident that the martian subsurface contains niche environments where life could develop. Key Words: Biomorph—Clays—Search for life (biosignatures)—Martian meteorites—Hydrothermal systems. Astrobiology 14, 651–693.
PMCID: PMC4126275  PMID: 25046549
17.  Utilization of Low-Pressure Plasma to Inactivate Bacterial Spores on Stainless Steel Screws 
Astrobiology  2013;13(7):597-606.
A special focus area of planetary protection is the monitoring, control, and reduction of microbial contaminations that are detected on spacecraft components and hardware during and after assembly. In this study, wild-type spores of Bacillus pumilus SAFR-032 (a persistent spacecraft assembly facility isolate) and the laboratory model organism B. subtilis 168 were used to study the effects of low-pressure plasma, with hydrogen alone and in combination with oxygen and evaporated hydrogen peroxide as a process gas, on spore survival, which was determined by a colony formation assay. Spores of B. pumilus SAFR-032 and B. subtilis 168 were deposited with an aseptic technique onto the surface of stainless steel screws to simulate a spore-contaminated spacecraft hardware component, and were subsequently exposed to different plasmas and hydrogen peroxide conditions in a very high frequency capacitively coupled plasma reactor (VHF-CCP) to reduce the spore burden. Spores of the spacecraft isolate B. pumilus SAFR-032 were significantly more resistant to plasma treatment than spores of B. subtilis 168. The use of low-pressure plasma with an additional treatment of evaporated hydrogen peroxide also led to an enhanced spore inactivation that surpassed either single treatment when applied alone, which indicates the potential application of this method as a fast and suitable way to reduce spore-contaminated spacecraft hardware components for planetary protection purposes. Key Words: Bacillus spores—Contamination—Spacecraft hardware—Plasma sterilization—Planetary protection. Astrobiology 13, 597–606.
PMCID: PMC3713438  PMID: 23768085
18.  A Potential Gravity-Sensing Role of Vascular Smooth Muscle Cell Glycocalyx in Altered Gravitational Stimulation 
Astrobiology  2013;13(7):626-636.
Previously, we have shown that vascular smooth muscle cells (VSMCs) exhibit varied physiological responses when exposed to altered gravitational conditions. In the present study, we focused on elucidating whether the cell surface glycocalyx could be a potential gravity sensor. For this purpose, a roller culture apparatus was used with the intent to provide altered gravitational conditions to cultured rat aortic smooth muscle cells (RASMCs). Heparinase III (Hep.III) was applied to degrade cell surface heparan sulfate proteoglycans (HSPG) selectively. Sodium chlorate was used to suppress new synthesis of HSPG. Glycocalyx remodeling, nitric oxide synthase (NOS) activation, and F-actin expression induced by gravity alteration were assessed by flow cytometry, reverse transcription polymerase chain reaction (RT-PCR), and Western blot. Results indicate that the exposure of cultured RASMCs to altered gravitational conditions led to a reduction in cell surface HSPG content and the activation of NOS. It also down-regulated the expression of glypican-1, constitutive NOS (NOSI and NOSIII), and F-actin. On the other hand, Hep.III followed by sodium chlorate treatment of HSPG attenuated the aforementioned NOS and F-actin modulation under altered gravitational conditions. All these findings suggest that the glycocalyx, and HSPG in particular, may be an important sensor of gravitational changes. This may play an important role in the regulation of NOS activation, F-actin modulation, and HSPG remodeling in VSMCs. Key Words: Glycocalyx—Gravity sensor—Gravity alteration—Roller culture apparatus. Astrobiology 13, 626–636.
PMCID: PMC3713443  PMID: 23848471
19.  Radiocarbon Evidence of Active Endolithic Microbial Communities in the Hyperarid Core of the Atacama Desert 
Astrobiology  2013;13(7):607-616.
The hyperarid core of the Atacama Desert is one of the driest and most inhospitable places on Earth, where life is most commonly found in the interior of rocks (i.e., endolithic habitats). Due to the extreme dryness, microbial activity in these habitats is expected to be low; however, the rate of carbon cycling within these microbial communities remains unknown. We address this issue by characterizing the isotopic composition (13C and 14C) of phospholipid fatty acids (PLFA) and glycolipid fatty acids (GLFA) in colonized rocks from four different sites inside the hyperarid core. δ13C results suggest that autotrophy and/or quantitative conversion of organic matter to CO2 are the dominant processes occurring with the rock. Most Δ14C signatures of PLFA and GLFA were consistent with modern atmospheric CO2, indicating that endoliths are using atmospheric carbon as a primary carbon source and are also cycling carbon quickly. However, at one site the PLFA contained 14C from atmospheric nuclear weapons testing that occurred during the 1950s and 1960s, indicating a decadal rate of carbon cycling. At the driest site (Yungay), based on the relative abundance and 14C content of GLFA and PLFA, there was evidence of possible preservation. Hence, in low-moisture conditions, glycolipids may persist while phospholipids are preferentially hydrolyzed. Key Words: Endoliths—Extremophile—Carbon isotopes—Radiocarbon—Lipids. Astrobiology 13, 607–616.
PMCID: PMC3713447  PMID: 23848470
20.  Reproductive and Locomotory Capacities of Caenorhabditis elegans Were Not Affected by Simulated Variable Gravities and Spaceflight During the Shenzhou-8 Mission 
Astrobiology  2013;13(7):617-625.
Reproduction and locomotion are essential features of animals that help to facilitate their interaction with the surrounding environment. Previous studies have produced inconsistent results on behavioral response to spaceflight by the model animal Caenorhabditis elegans (C. elegans) in liquid culture. Using standard agar-based nematode growth medium (NGM), we show here that both reproductive and locomotory capacities of C. elegans were not significantly changed by centrifuge-produced hypergravity or clinostat-simulated microgravity. To investigate the effect of actual spaceflight on C. elegans, a nematode test unit was specifically designed to maintain its normal growth on solid NGM slides and to allow automatic RNA fixation on board the Shenzhou-8 spaceflight. We did not detect alteration in either brood size of immediate progenies from postflight nematodes or locomotory behavior, including speed of locomotion, frequency of reversals, and rate of body bends of space-flown nematodes collected directly from nematode test units. Our results provide clear evidence that the nematode test unit is an appropriate apparatus for nematode growth on standard NGM and can be used for on-orbit analysis of C. elegans, including onboard RNA fixation for molecular analysis and real-time video acquisition for behavioral analysis, which are critical for further studies in unmanned spaceflight and outer space exploration. Key Words: Spaceflight—Hypergravity—Microgravity—Caenorhabditis elegans—Behavior—Reproduction. Astrobiology 13, 617–625.
PMCID: PMC3713449  PMID: 23837604
21.  Effect of SPE-like Proton or Photon Radiation on the Kinetics of Mouse Peripheral Blood Cells and Radiation Biological Effectiveness Determinations 
Astrobiology  2013;13(6):570-577.
Exploration missions outside low-Earth orbit are being planned; therefore, it is critical to understand the risk astronauts would be exposed to in the space environment, especially during extravehicular activities (EVAs). Reductions in white blood cell (WBC) numbers can occur as a result of exposure to solar particle event (SPE) radiation. The aim of the present study was to determine the duration of the effects on blood cell numbers from exposure to a single whole-body dose of SPE-like proton radiation or photon radiation as well as to determine the radiation biological effectiveness (RBE) values at those times when radiation exposure causes blood cell numbers to experience the most critical effects when using mice as a model. Our results indicate that both types of radiation cause significant reductions in the numbers of all blood cell types at different times post-irradiation. The RBE values were not significantly different from 1.0. These results indicate that the risk estimations for astronauts from exposure of mice to SPE-like proton radiation are comparable to those previously made for doses of standard reference radiations, suggesting that countermeasures should be developed for the decreases in blood cell counts observed following the exposure of mice to SPE radiation. Key Words: Proton radiation—Gamma radiation—Blood cell counts—Solar particle event. Astrobiology 13, 570–577.
PMCID: PMC3689168  PMID: 23980767
22.  Abiotic Production of Methane in Terrestrial Planets 
Astrobiology  2013;13(6):550-559.
On Earth, methane is produced mainly by life, and it has been proposed that, under certain conditions, methane detected in an exoplanetary spectrum may be considered a biosignature. Here, we estimate how much methane may be produced in hydrothermal vent systems by serpentinization, its main geological source, using the kinetic properties of the main reactions involved in methane production by serpentinization. Hydrogen production by serpentinization was calculated as a function of the available FeO in the crust, given the current spreading rates. Carbon dioxide is the limiting reactant for methane formation because it is highly depleted in aqueous form in hydrothermal vent systems. We estimated maximum CH4 surface fluxes of 6.8×108 and 1.3×109 molecules cm−2 s−1 for rocky planets with 1 and 5 M⊕, respectively. Using a 1-D photochemical model, we simulated atmospheres with volume mixing ratios of 0.03 and 0.1 CO2 to calculate atmospheric methane concentrations for the maximum production of this compound by serpentinization. The resulting abundances were 2.5 and 2.1 ppmv for 1 M⊕ planets and 4.1 and 3.7 ppmv for 5 M⊕ planets. Therefore, low atmospheric concentrations of methane may be produced by serpentinization. For habitable planets around Sun-like stars with N2-CO2 atmospheres, methane concentrations larger than 10 ppmv may indicate the presence of life. Key Words: Serpentinization—Exoplanets—Biosignatures—Planetary atmospheres. Astrobiology 13, 550–559.
PMCID: PMC3689174  PMID: 23742231
23.  Raman Spectroscopic Analysis of Geological and Biogeological Specimens of Relevance to the ExoMars Mission 
Astrobiology  2013;13(6):543-549.
A novel miniaturized Raman spectrometer is scheduled to fly as part of the analytical instrumentation package on an ESA remote robotic lander in the ESA/Roscosmos ExoMars mission to search for evidence for extant or extinct life on Mars in 2018. The Raman spectrometer will be part of the first-pass analytical stage of the sampling procedure, following detailed surface examination by the PanCam scanning camera unit on the ExoMars rover vehicle. The requirements of the analytical protocol are stringent and critical; this study represents a laboratory blind interrogation of specimens that form a list of materials that are of relevance to martian exploration and at this stage simulates a test of current laboratory instrumentation to highlight the Raman technique strengths and possible weaknesses that may be encountered in practice on the martian surface and from which future studies could be formulated. In this preliminary exercise, some 10 samples that are considered terrestrial representatives of the mineralogy and possible biogeologically modified structures that may be identified on Mars have been examined with Raman spectroscopy, and conclusions have been drawn about the viability of the unambiguous spectral identification of biomolecular life signatures. It is concluded that the Raman spectroscopic technique does indeed demonstrate the capability to identify biomolecular signatures and the mineralogy in real-world terrestrial samples with a very high degree of success without any preconception being made about their origin and classification. Key Words: Biosignatures—Mars Exploration Rovers—Raman spectroscopy—Search for life (biosignatures)—Planetary instrumentation. Astrobiology 13, 543–549.
PMCID: PMC3689185  PMID: 23758166
24.  Perchlorate Radiolysis on Mars and the Origin of Martian Soil Reactivity 
Astrobiology  2013;13(6):515-520.
Results from the Viking biology experiments indicate the presence of reactive oxidants in martian soils that have previously been attributed to peroxide and superoxide. Instruments on the Mars Phoenix Lander and the Mars Science Laboratory detected perchlorate in martian soil, which is nonreactive under the conditions of the Viking biology experiments. We show that calcium perchlorate exposed to gamma rays decomposes in a CO2 atmosphere to form hypochlorite (ClO−), trapped oxygen (O2), and chlorine dioxide (ClO2). Our results show that the release of trapped O2 (g) from radiation-damaged perchlorate salts and the reaction of ClO− with amino acids that were added to the martian soils can explain the results of the Viking biology experiments. We conclude that neither hydrogen peroxide nor superoxide is required to explain the results of the Viking biology experiments. Key Words: Mars—Radiolysis—Organic degradation—in situ measurement—Planetary habitability and biosignatures. Astrobiology 13, 515–520.
PMCID: PMC3691774  PMID: 23746165
25.  Survival of Organic Materials in Hypervelocity Impacts of Ice on Sand, Ice, and Water in the Laboratory 
Astrobiology  2014;14(6):473-485.
The survival of organic molecules in shock impact events has been investigated in the laboratory. A frozen mixture of anthracene and stearic acid, solvated in dimethylsulfoxide (DMSO), was fired in a two-stage light gas gun at speeds of ∼2 and ∼4 km s−1 at targets that included water ice, water, and sand. This involved shock pressures in the range of 2–12 GPa. It was found that the projectile materials were present in elevated quantities in the targets after impact and in some cases in the crater ejecta as well. For DMSO impacting water at 1.9 km s−1 and 45° incidence, we quantify the surviving fraction after impact as 0.44±0.05. This demonstrates successful transfer of organic compounds from projectile to target in high-speed impacts. The range of impact speeds used covers that involved in impacts of terrestrial meteorites on the Moon, as well as impacts in the outer Solar System on icy bodies such as Pluto. The results provide laboratory evidence that suggests that exogenous delivery of complex organic molecules from icy impactors is a viable source of such material on target bodies. Key Words: Organic—Hypervelocity—Shock—Biomarkers. Astrobiology 14, 473–485.
PMCID: PMC4060819  PMID: 24901745

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