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NASA NTRS2002

Reassessing the Evidence for the Earliest Traces of Life

The isotopic composition of graphite is commonly used as a biomarker in the oldest (>3.5 Gyr ago) highly metamorphosed terrestrial rocks. Earlier studies on isotopic characteristics of graphite occurring in rocks of the approximately 3.8-Gyr-old Isua supracrustal belt (ISB) in southern West Greenland have suggested the presence of a vast microbial ecosystem in the early Archean. This interpretation, however, has to be approached with extreme care. Here we show that graphite occurs abundantly in secondary carbonate veins in the ISB that are formed at depth in the crust by injection of hot fluids reacting with older crustal rocks (metasomatism). During these reactions, graphite forms from the disproportionation of Fe(II)-bearing carbonates at high temperature. These metasomatic rocks, which clearly lack biological relevance, were earlier thought to be of sedimentary origin and their graphite association provided the basis for inferences about early life. The new observations thus call for a reassessment of previously presented evidence for ancient traces of life in the highly metamorphosed Early Archaean rock record.

vanZullen, Mark A.↗
NASA NTRS2003

Siliceous Shrubs in Yellowstone's Hot Springs: Implications for Exobiological Investigations

Potential relict hot springs have been identified on Mars and, using the Earth as an analog, Martian hot springs are postulated to be an optimal locality for recognizing preserved evidence of extraterrestrial life. Distinctive organic and inorganic biomarkers are necessary to recognize preserved evidence of life in terrestrial and extraterrestrial hot spring accumulations. Hot springs in Yellowstone National Park, Wyoming, U.S.A., contain a wealth of information about primitive microbial life and associated biosignatures that may be useful for future exobiological investigations. Numerous siliceous hot springs in Yellowstone contain abundant, centimeter-scale, spinose precipitates of opaline silica (opal-A). Although areally extensive in siliceous hot spring discharge channel facies, these spinose forms have largely escaped attention. These precipitates referred to as shrubs, consist of porous aggregates of spinose opaline silica that superficially resemble miniature woody plants, i.e., the term shrubs. Shrubs in carbonate precipitating systems have received considerable attention, and represent naturally occurring biotically induced precipitates. As such, shrubs have great potential as hot spring environmental indicators and, more importantly, proxies for pre-existing microbial life.

S A Guidry↗
NASA NTRS2003

Morphological Evidence for an Exclusively Inorganic Origin for Magnetite in Martian Meteorite ALH84001

The origin of magnetite crystals in Martian Meteorite ALH84001 is the focus of a debate about the possibility of past (and present) life on Mars. McKay et al. originally suggested that some of the magnetite crystals associated with carbonate globules in Martian Meteorite ALH84001 are biogenic in ori-gin, because they are single magnetic domain, free of crystalline defects, chemically pure, and coexist with other metastable phases in apparent disequilibrium. Thomas-Keprta et al. reported that a subpopulation of magnetite crystals (approx. 25%) associated with carbonate globules in ALH84001 and magnetite crystals produced by magnetotactic bacterial strain MV-1 have similar morphologies with crystal elongation along the [111] crystallographic axis that they describe as "truncated hexa-octahedral" ([111-THO]) magnetite. Along with several other properties, the [111]-THO morphology has been proposed to constitute a biomarker (i.e., formed only in biogenic processes), so that the presence of [111]-THO magnetite in ALH84001 may be evidence for past life on Mars.

Golden, D. C.↗
NASA NTRS2003

Magnetic Analysis Techniques Applied to Desert Varnish

Desert varnish is a black or reddish coating commonly found on rock samples from arid regions. Typically, the coating is very thin, less than half a millimeter thick. Previous research has shown that the primary components of desert varnish are silicon oxide clay minerals (~60%), manganese and iron oxides (~20-30%), and trace amounts of other compounds [1]. Desert varnish is thought to originate when windborne particles containing iron and manganese oxides are deposited onto rock surfaces where manganese oxidizing bacteria concentrate the manganese and form the varnish [4,5]. If desert varnish is indeed biogenic, then the presence of desert varnish on rock surfaces could serve as a biomarker, indicating the presence of microorganisms. This idea has considerable appeal, especially for Martian exploration [6]. Magnetic analysis techniques have not been extensively applied to desert varnish. The only previous magnetic study reported that based on room temperature demagnetization experiments, there were noticeable differences in magnetic properties between a sample of desert varnish and the substrate sandstone [7]. Based upon the results of the demagnetization experiments, the authors concluded that the primary magnetic component of desert varnish was either magnetite (Fe3O4) or maghemite ( Fe2O3).

Schmidgall, E. R.↗
NASA NTRS2003

Microfossils in CI and CO Carbonaceous Meteorites

Secondary and backscatter electron images and x-ray spectral data of selected CI (Alais, Orgueil, and Tagish Lake) and CO3 (Rainbow and Dar a1 Gani 749) carbonaceous meteorites have recently been obtained using Field Emission and Environmental Scanning Electron Microscopes These studies indicate the presence of a large assemblage of biomarkers and complex lithified and carbonized remains of bodies that we interpret as indigenous microfossils. We discuss the meteorites, provide images of many of the biogenic forms found embedded in the freshly fractured meteorite surfaces.

Hoover, Richard B.↗
NASA NTRS2004

Signs of Life in Meridiani Planum: What Might Opportunity See (Or Miss)?

The Meridiani Planum hematite deposit has been designated as the prime landing site for Opportunity, one of the two Mars Exploration Rover (MER) spacecraft. Iron oxide and hydroxide minerals, including hematite, can mineralize and preserve microfossils and physical biomarkers. Previous research by ourselves and others, summarized below, has demonstrated such mineralization in a variety of terrestrial hematite deposits. Our most recent study has focused on the mineralization of microorganisms in rock varnish, the iron- and manganese-rich layers that coat rocks in many arid environments.

Allen, C. C.↗
NASA NTRS2000

Quantifying the Intercontinental and Global Reach and Effects of Pollution

The Atmospheric Chemistry Modeling Group is participating in an international effort to explore the projected interactions of the atmosphere with biota, human activity, and the natural environment over the next three decades. The group uses computer simulations and statistical analyses to compare theory and observations of the composition of the lower atmosphere. This study of global habitability change is part of a more ambitious activity to understand global habitability. This broad planetary understanding is central to planetary habitability, biomarker detection, and similar aspects of Astrobiology. The group has made highly detailed studies of immense intercontinental plumes that affect the chemistry of the global atmosphere, especially the region below the ozone (O3) layer whose chemical composition defines the conditions for healthy humans and the biosphere. For some decades there has been concern about the pollution from cities and industrial burning and its possible effect in increasing smog ozone, not only in continental regions, but also in plumes that spread downwind. Recently, there has been new concern about another kind of pollution plume. Projections for a greatly expanded aircraft fleet imply that there will be plumes of nitrogen oxides (NO(x)) from jet exhaust in the Northern Hemisphere downwind of major air traffic routes. Both of these are tied to large-scale O3 in the troposphere, where it is toxic to humans and plant tissues.

Chatfield, Robert B.↗
NASA NTRS2004

Acquisition of a Gas Chromatograph/Mass Spectrometer System for Laboratory Study of Prebiotic Organic Geochemical Processes on the Early Earth, Mars, and Meteorites

This was a major equipment grant that provided funds ($72K) for purchase of a benchtop gas chromatograph-mass spectrometer (GC-MS) for use in experimental studies of prebiotic organic compounds. An Agilent model 689015973 GC-MS was purchased and installed in the PI's lab in August of 2003. The instrument is now being used for a variety of research projects. The primary use of the instrument is to analyze and quantify organic products of laboratory experiments conducted by the PI. One example is shown, which shows organic products (predominantly n-alkanes) formed during Fischer-Tropsch-type abiotic synthesis under hydrothermal conditions. The analytical capabilities of the GC- MS allowed identification of the numerous organic products of this as well as other laboratory experiments. A key use of the instrument in this research is that the mass spectrometer capabilities allow use of isotopically labeled reactants to trace the progress of reactions and evaluate background contaminants. collaborative projects with other scientists involved in exobiology & astrobiology research (e.g., Mitch Schulte, NASA Ames; Katrina Edwards, Woods Hole Oceanographic Institution). For instance, an analysis of membrane lipids of an lithoautotrophic iron-oxidizing bacteria being grown on basalt as a source of metabolic energy, a project where the instrument is being used to evaluate possible biomarker compounds from these organisms is shown. These iron oxidizers are thought to be similar to those living within the ocean crust, and are being investigated as possible analog organisms to those on the early Earth or crust of Mars. The instrument has also been used by an outside investigator (graduate student Brandon Canfeld, Arizona State University) for identification and isotopic characterization of experimental products of abiotic organic synthesis experiments he is conducting with Dr. John Holloway. analysis of quality control samples for other NASA-funded projects. For instance, an analysis of residual hydrocarbon contaminants on the internal surface of the shell of an atmospheric sounding rocket is shown. This analysis was used to help determine the source of the contaminating compounds. In the future, the instrument will continue to be used for quality control analysis in clean rooms and instrument construction facilities within the Laboratory for Atmospheric and Space Physics, where the GC-MS is housed.

McCollom, Thomas↗
NASA NTRS2004

Enzyme Amplified Detection of Microbial Cell Wall Components

This proposal is MBL's portion of NASA's Johnson Space Center's Astrobiology Center led by Principal Investigator, Dr. David McKay, entitled: 'Institute for the Study of Biomarkers in Astromaterials.' Dr. Norman Wainwright is the principal investigator at MBL and is responsible for developing methods to detect trace quantities of microbial cell wall chemicals using the enzyme amplification system of Limulus polyphemus and other related methods.

Norman R Wainwright↗
NASA NTRS2003

Cyclooxygenase-2 expression is related to nuclear grade in ductal carcinoma in situ and is increased in its normal adjacent epithelium

Cyclooxygenase-2 (COX-2) is emerging as an important cancer biomarker and is now an experimental target for solid tumor treatment.However, no study has exclusively focused on COX-2 expression in early lesions such as ductal carcinoma in situ (DCIS). We examined COX-2 expression by immunohistochemistry in 46 cases of women undergoing surgical resection for DCIS. We found that COX-2 expression was detected in 85% of all DCIS specimens, with increased COX-2 staining correlating with higher nuclear grade. Strikingly, COX-2 staining intensity in the normal adjacent epithelium was stronger than in the DCIS lesion itself. Our observations demonstrate that COX-2 is up-regulated in the normal adjacent epithelium and supports the hypothesis that the surrounding epithelial tissue is part of the disease process in DCIS.

NASA Discipline Radiation Health↗
NASA NTRS1992

Novel long-chain anteiso-alkanes and anteiso-alkanoic acids in Antarctic rocks colonized by living and fossil cryptoendolithic microorganisms

Saponified extracts of rock samples colonized by cryptoendolithic microbial communities from the McMurdo Dry Valleys of Southern Victoria Land, Antarctica, were separated into hydrocarbon and fatty acid fractions by silica gel column chromatography. Hydrocarbons and methyl esters of fatty acids were analyzed by capillary gas chromatography-mass spectrometry. Unusually, a suite of long-chain anteiso-alkanes (a-C20 to a-C30) and anteiso-alkanoic acids (a-C20 to a-C30) were detected in many samples, together with straight-chain, branched and/or cyclic and acyclic isoprenoid compounds. These novel compounds are probably derived from unidentified heterotrophic bacteria or symbiotic processes in a unique microbial community in the Antarctic cold desert and suggest the occurrence of a special biosynthetic pathway. Long-chain anteiso-alkanes are probably formed through microbial decarboxylation of corresponding anteiso-alkanoic acids. They may serve as new biomarkers in environmental and geochemical studies.

NASA Program Exobiology↗
NASA NTRS1994

Complex polar lipids of a hot spring cyanobacterial mat and its cultivated inhabitants

The complex polar lipids of the hot spring cyanobacterial mat in the 50 to 55 degrees C region of Octopus Spring, Yellowstone National Park, and of thermophilic bacteria cultivated from this or similar habitats, were compared in an attempt to understand the microbial sources of the major lipid biomarkers in this community. Intact complex lipids were analyzed directly by fast atom bombardment mass spectrometry (FAB-MS), two-dimensional thin-layer chromatography (TLC), and combined TLC-FAB-MS. FAB-MS and TLC gave qualitatively similar results, suggesting that the mat contains major lipids most like those of the cyanobacterial isolate we studied, Synechococcus sp. strain Y-7c-s. These include monoglycosyl, diglycosyl, and sulfoquinosovyl diglycerides (MG, DG, and SQ, respectively) and phosphatidyl glycerol (PG). Though Chloroflexus aurantiacus also contains MG, DG, and PG, the fatty acid chain lengths of mat MGs, DGs, and PGs resemble more those of cyanobacterial than green nonsulfur bacterial lipids. FAB-MS spectra of the lipids of nonphototrophic bacterial isolates were distinctively different from those of the mat and phototrophic isolates. The lipids of these nonphototrophic isolates were not detected in the mat, but most could be detected when added to mat samples. The mat also contains major glycolipids and aminophospholipids of unknown structure and origin. FAB-MS and TLC did not always give quantitatively similar results. In particular, PG and SQ may give disproportionately high FAB-MS responses.

Non-NASA Center↗
NASA NTRS2004

Microbial monitoring of spacecraft and associated environments

Rapid microbial monitoring technologies are invaluable in assessing contamination of spacecraft and associated environments. Universal and widespread elements of microbial structure and chemistry are logical targets for assessing microbial burden. Several biomarkers such as ATP, LPS, and DNA (ribosomal or spore-specific), were targeted to quantify either total bioburden or specific types of microbial contamination. The findings of these assays were compared with conventional, culture-dependent methods. This review evaluates the applicability and efficacy of some of these methods in monitoring the microbial burden of spacecraft and associated environments. Samples were collected from the surfaces of spacecraft, from surfaces of assembly facilities, and from drinking water reservoirs aboard the International Space Station (ISS). Culture-dependent techniques found species of Bacillus to be dominant on these surfaces. In contrast, rapid, culture-independent techniques revealed the presence of many Gram-positive and Gram-negative microorganisms, as well as actinomycetes and fungi. These included both cultivable and noncultivable microbes, findings further confirmed by DNA-based microbial detection techniques. Although the ISS drinking water was devoid of cultivable microbes, molecular-based techniques retrieved DNA sequences of numerous opportunistic pathogens. Each of the methods tested in this study has its advantages, and by coupling two or more of these techniques even more reliable information as to microbial burden is rapidly obtained. Copyright 2004 Springer-Verlag.

Environmental Monitoring/methods↗
NASA NTRS2005

Influence of Planetary Protection Guidelines on Waste Management Operations

Newly outlined missions in the Space Exploration Initiative include extended human habitation on Mars. During these missions, large amounts of waste materials will be generated in solid, liquid and gaseous form. Returning these wastes to Earth will be extremely costly, and will therefore likely remain on Mars. Untreated, these wastes are a reservoir of live/dead organisms and molecules considered to be "biomarkers" i.e., indicators of life). If released to the planetary surface, these materials can potentially confound exobiology experiments and disrupt Martian ecology indefinitely (if existent). Waste management systems must therefore be specifically designed to control release of problematic materials both during the active phase of the mission, and for any specified post-mission duration. To effectively develop waste management requirements for Mars missions, planetary protection guidelines must first be established. While previous policies for Apollo lunar missions exist, it is anticipated that the increased probability of finding evidence of life on Mars, as well as the lengthy mission durations will initially lead to more conservative planetary protection measures. To facilitate the development of overall requirements for both waste management and planetary protection for future missions, a workshop was conducted to identify how these two areas interface, and to establish a preliminary set of planetary protection guidelines that address waste management operations. This paper provides background regarding past and current planetary protection and waste management issues, and their interactions. A summary of the recommended planetary protection guidelines, anticipated ramifications and research needs for waste management system design for both forward (Mars) and backward (Earth) contamination is also provided.

Hogan, John A.↗
NASA NTRS2005

A Compact Instrument for Remote Raman and Fluorescence Measurements to a Radial Distance of 100 m

Compact remote spectroscopic instruments that could provide detailed information about mineralogy, organic and biomaterials on a planetary surface over a relatively large area are desirable for NASA s planetary exploration program. Ability to explore a large area on the planetary surfaces as well as in impact craters from a fixed location of a rover or lander will enhance the probability of selecting target rocks of high scientific contents as well as desirable sites in search of organic compounds and biomarkers on Mars and other planetary bodies. We have developed a combined remote inelastic scattering (Raman) and laser-induced fluorescence emission (LIFE) compact instrument capable of providing accurate information about minerals, organic and biogenic materials to a radial distance of 100 m. Here we present the Raman and LIFE (R-LIFE) data set.

Sharma, S. K.↗
NASA NTRS2004

Simultaneous measurement of multiple radiation-induced protein expression profiles using the Luminex(TM) system

Space flight results in the exposure of astronauts to a mixed field of radiation composed of energetic particles of varying energies, and biological indicators of space radiation exposure provides a better understanding of the associated long-term health risks. Current methods of biodosimetry have employed the use of cytogenetic analysis for biodosimetry, and more recently the advent of technological progression has led to advanced research in the use of genomic and proteomic expression profiling to simultaneously assess biomarkers of radiation exposure. We describe here the technical advantages of the Luminex(TM) 100 system relative to traditional methods and its potential as a tool to simultaneously profile multiple proteins induced by ionizing radiation. The development of such a bioassay would provide more relevant post-translational dynamics of stress response and will impart important implications in the advancement of space and other radiation contact monitoring. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

NASA Center JSC↗
NASA NTRS2005

Comets, Carbonaceous Meteorites, and the Origin of the Biosphere

The biosphere comprises the Earth s crust, atmosphere, oceans, and ice caps and the living organisms that survive within this habitat. The discoveries of barophilic chemolithoautotrophic thermophiles living deep within the crust and in deep-sea hydrothermal vents, and psychrophiles in permafrost and deep within the Antarctic Ice Sheet indicate the Earth s biosphere is far more extensive than previously recognized. Molecular biomarkers and Bacterial Paleontology provide evidence that life appeared very early on the primitive Earth and the origin of the biosphere is closely linked with the emergence of life. The role of comets, meteorites, and interstellar dust in the delivery of water, organics and prebiotic chemicals has long been recognized. Deuterium enrichment of seawater and comets indicates that comets delivered oceans to the early Earth. Furthermore, the similarity of the D/H ratios and the chemical compositions of CI carbonaceous meteorites and comets indicate that the CI meteorites may be remnants of cometary nuclei with most volatiles removed. Comets, meteorites, and interstellar dust also contain complex organic chemicals, amino acids, macromolecules, and kerogen-like biopolymers and may have played a crucial role in the delivery of complex organics and prebiotic chemicals during the Hadean (4.5-3.8 Gyr) period of heavy bombardment. The existence of indigenous microfossils of morphotypes of cyanobacteria in the CI and CM carbonaceous meteorites suggests that the paradigm that life originated endogenously in the primitive oceans of early Earth may require re-consideration. Recent data on the hot (300-400 K) black crust on comet P/Halley and Stardust images of P/Wild 2 showing depressions, tall cliffs, and pinnacles, indicate the presence of thick, durable, dark crusts on comets. If cavities within the ice and crust sustain vapor pressures in excess of 10 millibar, then localized pools of liquid water and brines could exist within the comet. Since life exists on Earth wherever there is liquid water, it is suggested that comets might also harbour viable and/or cryopreserved microbiota.

Hoover, Richard B.↗
NASA NTRS2005

Morphology and Elemental Composition of Recent and Fossil Cyanobacteria

Cyanobacteria (cyanophyta, cyanoprokaryota, and blue-green algae) are an ancient, diverse and abundant group of photosynthetic oxygenic microorganisms. Together with other bacteria and archaea, the cyanobacteria have been the dominant life forms on Earth for over 3.5 billion years. Cyanobacteria occur in some of our planets most extreme environments - hot springs and geysers, hypersaline and alkaline lakes, hot and cold deserts, and the polar ice caps. They occur in a wide variety of morphologies. Unlike archaea and other bacteria, which are typically classified in pure culture by their physiological, biochemical and phylogenetic properties, the cyanobacteria have historically been classified based upon their size and morphological characteristics. These include the presence or absence of heterocysts, sheath, uniseriate or multiseriate trichomes, true or false branching, arrangement of thylakoids, reproduction by akinetes, binary fission, hormogonia, fragmentation, presence/absence of motility etc. Their antiquity, distribution, structural and chemical differentiation, diversity, morphological complexity and large size compared to most other bacteria, makes the cyanobacteria ideal candidates for morphological biomarkers in returned Astromaterials. We have obtained optical (nomarski and phase contrast)/fluorescent (blue and green excitation) microscopy images using an Olympus BX60 compound microscope and Field Emission Scanning Electron Microscopy images and EDAX elemental compositions of living and fossil cyanobacteria. The S-4000 Hitachi Field Emission Scanning Electron Microscope (FESEM) has been used to investigate microfossils in freshly fractured interior surfaces of terrestrial rocks and the cells, hormogonia, sheaths and trichomes of recent filamentous cyanobacteria. We present Fluorescent and FESEM Secondary and Backscattered Electron images and associated EDAX elemental analyses of recent and fossil cyanobacteria, concentrating on representatives of the genera Calothnx, Leptolyngbya, Lyngbya, Planktolyngbya and Oscillatoria.

SaintAmand, Ann↗