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Increased genome instability in Escherichia coli lon mutants: relation to emergence of multiple-antibiotic-resistant (Mar) mutants caused by insertion sequence elements and large tandem genomic amplifications.

Thirteen spontaneous multiple-antibiotic-resistant (Mar) mutants of Escherichia coli AG100 were isolated on Luria-Bertani (LB) agar in the presence of tetracycline (4 microg/ml). The phenotype was linked to insertion sequence (IS) insertions in marR or acrR or unstable large tandem genomic amplifications which included acrAB and which were bordered by IS3 or IS5 sequences. Five different lon mutations, not related to the Mar phenotype, were also found in 12 of the 13 mutants. Under specific selective conditions, most drug-resistant mutants appearing late on the selective plates evolved from a subpopulation of AG100 with lon mutations. That the lon locus was involved in the evolution to low levels of multidrug resistance was supported by the following findings: (i) AG100 grown in LB broth had an important spontaneous subpopulation (about 3.7x10(-4)) of lon::IS186 mutants, (ii) new lon mutants appeared during the selection on antibiotic-containing agar plates, (iii) lon mutants could slowly grow in the presence of low amounts (about 2x MIC of the wild type) of chloramphenicol or tetracycline, and (iv) a lon mutation conferred a mutator phenotype which increased IS transposition and genome rearrangements. The association between lon mutations and mutations causing the Mar phenotype was dependent on the medium (LB versus MacConkey medium) and the antibiotic used for the selection. A previously reported unstable amplifiable high-level resistance observed after the prolonged growth of Mar mutants in a low concentration of tetracycline or chloramphenicol can be explained by genomic amplification.

Anti-Bacterial Agents↗

Active efflux of chloramphenicol in susceptible Escherichia coli strains and in multiple-antibiotic-resistant (Mar) mutants.

The multiple-antibiotic resistance (mar) locus (min 34) regulates a resistance to chloramphenicol in Escherichia coli that does not involve acetyltransferase. Transport studies showed that wild-type cells had an apparent endogenous active efflux of chloramphenicol which depended on the proton motive force. This efflux was not altered by a 39-kb chromosomal deletion which included the mar locus. Nevertheless, mutations at the mar locus led to a stronger net chloramphenicol efflux. Therefore, a gene encoding the putative efflux system cannot be at the mar locus but may be positively influenced by that locus.

Cell Membrane↗

Characterization of MarR, the repressor of the multiple antibiotic resistance (mar) operon in Escherichia coli.

The marRAB operon is one of two operons in the mar locus of Escherichia coli that are divergently transcribed from a central regulatory region, marO. The marRAB operon, transcribed from marOII, controls intrinsic resistance or susceptibility to multiple antibiotics and is inducible by structurally unrelated compounds such as tetracycline and chloramphenicol (S. P. Cohen, H. Hachler, and S. B. Levy, J. Bacteriol. 175:1484-1492, 1993). To clarify the role of the operon in response to environmental signals, its transcription was studied under different conditions, using a marOII-lacZ transcriptional fusion introduced into the chromosome of wild-type or mar-deleted cells. In wild-type cells, uncoupling agents (such as carbonyl cyanide m-chlorophenylhydrazone) and different redox-cycling compounds (e.g., menadione and plumbagin) induced expression from the marOII-lacZ fusion two- to sevenfold. In the mar-deleted strain, LacZ expression from the fusion was 10-fold higher than in wild-type cells. This activity was temperature sensitive (3-fold lower at 42 than at 30 degrees C) and decreased 20-fold with the introduction of the gene for MarR. Structurally different compounds which induce the mar operon in wild-type cells reversed the MarR repression of marOII-lacZ expression. To determine the size of MarR, it was fused to MalE as a MarR fusion protein of 144 amino acids [MarR(144)] or of 125 amino acids (deleted of 19 amino acids at the N terminus) [MarR(125)]. Only the MarR(144) fusion showed repressor ability. The purified MarR(144) fusion, but not the MarR(125) fusion, bound specifically to marO in vitro, as revealed by gel retardation, with an apparent dissociation constant of 5 x 10(-9) M. MarR, therefore, controls expression of the marRAB operon presumably by binding to marO. MarR repression in cells can be reversed by different compounds, facilitating the response of bacteria to multiple environmental stress conditions.

Bacterial Proteins↗

Fis, an accessorial factor for transcriptional activation of the mar (multiple antibiotic resistance) promoter of Escherichia coli in the presence of the activator MarA, SoxS, or Rob.

Transcription of the multiple antibiotic resistance marRAB operon increases when one of the sequence-related activators, MarA, SoxS, or Rob, binds to the "marbox" centered at -61.5 relative to the transcriptional start site. Previous deletion analyses showed that an adjacent upstream "accessory region" was needed to augment the marbox-dependent activation. To analyze the roles of the marbox and accessory regions on mar transcription, thirteen promoters, each with a different 5-bp transversion of the -96 to -32 sequence, were synthesized, fused to lacZ, and assayed for beta-galactosidase production in single-copy lysogens with appropriate genotypes. The accessory region is shown here to be a binding site for Fis centered at -81 and to bind Fis, a small DNA-binding and -bending protein, with a Kd of approximately 5 nM. The binding of MarA to the marbox and that of Fis to its site were independent of each other. MarA, SoxS, and Rob each activated the mar promoter 1.5-to 2-fold when it had a wild-type marbox but Fis was absent. In the presence of MarA, SoxS, or Rob, Fis further enhanced the activity of the promoter twofold provided the promoter was also capable of binding Fis. However, in the absence of MarA, SoxS, or Rob or in the absence of a wild-type marbox, Fis nonspecifically lowered the activity of the mar promoter about 25% whether or not a wild-type Fis site was present. Thus, Fis acts as an accessory transcriptional activator at the mar promoter.

Bacterial Proteins↗

The Salmonella typhimurium mar locus: molecular and genetic analyses and assessment of its role in virulence.

The marRAB operon is a regulatory locus that controls multiple drug resistance in Escherichia coli. marA encodes a positive regulator of the antibiotic resistance response, acting by altering the expression of unlinked genes. marR encodes a repressor of marRAB transcription and controls the production of MarA in response to environmental signals. A molecular and genetic study of the homologous operon in Salmonella typhimurium was undertaken, and the role of marA in virulence in a murine model was assessed. Expression of E. coli marA (marAEC) present on a multicopy plasmid in S. typhimurium resulted in a multiple antibiotic resistance (Mar) phenotype, suggesting that a similar regulon exists in this organism. A genomic plasmid library containing S. typhimurium chromosomal sequences was introduced into an E. coli strain that was deleted for the mar locus and contained a single-copy marR'-'lacZ translational fusion. Plasmid clones that contained both S. typhimurium marR (marRSt) and marA (marASt) genes were identified as those that were capable of repressing expression of the fusion and which resulted in a Mar phenotype. The predicted amino acid sequences of MarRSt, MarASt, and MarBSt were 91, 86, and 42% identical, respectively, to the same genes from E. coli, while the operator/promoter region of the operon was 86% identical to the same 98-nucleotide-upstream region in E. coli. The marRAB transcriptional start sites for both organisms were determined by primer extension, and a marRABSt transcript of approximately 1.1 kb was identified by Northern blot analysis. Its accumulation was shown to be inducible by sodium salicylate. Open reading frames flanking the marRAB operon were also conserved. An S. typhimurium marA disruption strain was constructed by an allelic exchange method and compared to the wild-type strain for virulence in a murine BALB/c infection model. No effect on virulence was noted. The endogenous S. typhimurium plasmid that is associated with virulence played no role in marA-mediated multiple antibiotic resistance. Taken together, the data show that the S. typhimurium mar locus is structurally and functionally similar to marRABEc and that a lesion in marASt has no effect on S. typhimurium virulence for BALB/c mice.

Amino Acid Sequence↗

Positron emission tomographic analysis of central dopamine D1 receptor binding in normal subjects treated with the atypical neuroleptic, SDZ MAR 327.

SDZ MAR 327 is a new neuroleptic agent with high in vitro affinity for dopamine D1 and D2 receptors. The goal of this study was to determine the effect of time after SDZ MAR 327 administration on central dopamine D1 receptor occupancy in healthy humans. Positron emission tomography (PET) with the dopamine D1 receptor ligand, [11C] SCH 23390, was performed in 6 male volunteers (age 22-34), in both the drug naive state and at 1, 2 and 4 h after a single oral dose of SDZ MAR 327 (9 mg). The pre and post drug treatment [11C] SCH 23390 dynamic data were analyzed using two different methods, each yielding a parameter proportional to the receptor density: i) a simple regional comparison approximating the specifically bound to free fraction, B/F; and ii) a two compartment, two parameter model yielding the apparent distribution volume DV". With both methods, a metabolite corrected arterial input function was used and the vascular fraction of tissue (Vb) was fixed at a previously determined value of 4%. Method I served as a qualitative comparison of the paired studies and demonstrated little difference between the pre and post drug conditions, method II also confirmed that there was no significant change in binding of [11C] SCH 23390 in the striatum. These data indicate that SDZ MAR 327 produces little if any effect on dopamine D1 receptor binding at the dose used.

Adult↗

Inflammatory Syndromes (SIRS, MARS, CARS) in Patients with Surgical Infection.

In the present study 37 patients with surgical infection were investigated and a new set of diagnostic tests for detection of major syndromes of systemic inflammation - systemic inflammatory response syndrome (SIRS), compensatory anti-inflammatory response syndrome (CARS) and mixed antagonist response syndrome (MARS) - was developed. In summary, we have demonstrated that patients with surgical infection were characterized by an immunodeficiency with significant reduction of mitogen-induced proliferation and IL-2/IL-4 production in vitro combined with decrease of HLA DR(+) monocytes. Furthermore, it was revealed that the patient's serum exhibited substantially enhanced suppressive and inflammatory activities as well as the level of C-reactive protein. We have defined the negative correlation between the serum inflammatory and suppressive activities (SIA and SSA) that was most prominent at the early stage of disease. Since the changes of serum bioactivity in the course of surgical infection were prominent and coherent, we supposed that tested activities might reflect the distinctive features of systemic inflammation. In according to this assumption, the patients were divided into 3 subgroups with predominance of SIRS, CARS and MARS by using the SIA and SSA expression. It has been shown that SIRS was more frequently detected at the early stage, whereas CARS - at the late stage of disease. Patients with SIRS, CARS or MARS significantly differed by the content of CD8(+) T and CD72(+) B lymphocytes, the concentration of IgG and IgA, the production of IL-2 and IL-4. Finally, the data obtained from patients, those were studied repeatedly, showed the possibility of transformation of the major systemic inflammatory syndromes during the disease course. Our findings suggest that measurement of serum inflammatory and suppressive activities may help to differentiate patients with SIRS, CARS or MARS and to select the appropriate strategy of immunotherapy.

Journal Article↗

Successful treatment of an adult with Amanita phalloides-induced fulminant liver failure with molecular adsorbent recirculating system (MARS).

Despite significant advances in intensive care management of Amanita phalloides-induced fulminant liver failure (FLF), patients with this condition still have a high mortality rate in the absence of orthotopic liver transplantation. Molecular Adsorbent Recirculating System (MARS) is a new, cell-free, extracorporeal liver assistance method utilizing an albumin dialysate for the removal of albumin-bound toxins, and a highly effective depurative therapy in adults with wild mushroom-induced FLF. We report the case of a 39 year old woman with Amanita phalloides-induced FLF, admitted to our intensive care unit (ICU) and treated with MARS. Our patient had severe hepatic dysfunction: hepatic encephalopathy (grade II), ALT = 5022 (2475-10098) IU/L, bilirubin = 7.18 (4.8-10.1) mg/dL, prothrombin time (PT) = 90.4 (29.3-140.4) s. MARS sessions had an immediate impact on liver tests: statistically significant decrease in ammonia, ALT and PT. Hepatic encephalopathy was successfully reduced. The patient survived and the hepatic function completely recovered. MARS appears to be a safe and highly effective depurative therapy in adults with Amanita phalloides-induced FLF.

Adult↗

Immunoglobulin kappa gene expression after stable integration. I. Role of the intronic MAR and enhancer in plasmacytoma cells.

Rearranged MOPC41 immunoglobulin kappa gene constructs have been stably introduced into cultured S194 mouse plasmacytoma cells to investigate the effects of deleting the intronic enhancer and/or matrix association region (MAR) on gene expression. Intact single-copy kappa genes containing 1.5 kilobase pairs of upstream and 8.5 kilobase pairs of downstream flanking sequences exhibited sensitivity to chromosome position effects and were expressed at a mean level of 27% relative to the endogenous kappa gene expression or only 6% with respect to the MOPC41 kappa mRNA levels in the tumor. Deletion of the intronic MAR led to a 4-fold decrease in expression, while deletion of both the MAR and enhancer led to an 11-fold decline. These effects were dampened by preselecting for integration into a transcriptionally poised chromatin location as demonstrated by linkage to a selectable marker which lacked both a MAR and an enhancer. Significantly, we found that sequences downstream of the poly(A) addition site compensated 150-fold for deletion of the intronic enhancer.

Animals↗

Neobacillus driksii sp. nov. isolated from a Mars 2020 spacecraft assembly facility and genomic potential for lasso peptide production in Neobacillus.

UNLABELLED: During microbial surveillance of the Mars 2020 spacecraft assembly facility, two novel bacterial strains, potentially capable of producing lasso peptides, were identified. Characterization using a polyphasic taxonomic approach, whole-genome sequencing and phylogenomic analyses revealed a close genetic relationship among two strains from Mars 2020 cleanroom floors (179-C4-2-HS, 179-J1A1-HS), one strain from the Agave plant (AT2.8), and another strain from wheat-associated soil (V4I25). All four strains exhibited high 16S rRNA gene sequence similarity (>99.2%) and low average nucleotide identity (ANI) with Neobacillus niacini NBRC 15566T, delineating new phylogenetic branches within the genus. Detailed molecular analyses, including gyrB (90.2%), ANI (86.4%), average amino acid identity (87.8%) phylogenies, digital DNA-DNA hybridization (32.6%), and percentage of conserved proteins (77.7%) indicated significant divergence from N. niacini NBRC 15566T. Consequently, these strains have been designated Neobacillus driksii sp. nov., with the type strain 179-C4-2-HST (DSM 115941T = NRRL B-65665T). N. driksii grew at 4°C to 45°C, pH range of 6.0 to 9.5, and 0.5% to 5% NaCl. The major cellular fatty acids are iso-C15:0 and anteiso-C15:0. The dominant polar lipids include diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified aminolipid. Metagenomic analysis within NASA cleanrooms revealed that N. driksii is scarce (17 out of 236 samples). Genes encoding the biosynthesis pathway for lasso peptides were identified in all N. driksii strains and are not commonly found in other Neobacillus species, except in 7 out of 26 recognized species. This study highlights the unique metabolic capabilities of N. driksii, underscoring their potential in antimicrobial research and biotechnology. IMPORTANCE: The microbial surveillance of the Mars 2020 assembly cleanroom led to the isolation of novel N. driksii with potential applications in cleanroom environments, such as hospitals, pharmaceuticals, semiconductors, and aeronautical industries. N. driksii genomes were found to possess genes responsible for producing lasso peptides, which are crucial for antimicrobial defense, communication, and enzyme inhibition. Isolation of N. driksii from cleanrooms, Agave plants, and dryland wheat soils, suggested niche-specific ecology and resilience under various environmentally challenging conditions. The discovery of potent antimicrobial agents from novel N. driksii underscores the importance of genome mining and the isolation of rare microorganisms. Bioactive gene clusters potentially producing nicotianamine-like siderophores were found in N. driksii genomes. These siderophores can be used for bioremediation to remove heavy metals from contaminated environments, promote plant growth by aiding iron uptake in agriculture, and treat iron overload conditions in medical applications.

Phylogeny↗

A coupled soil-atmosphere model of H2O2 on Mars.

The Viking Gas Chromatograph Mass Spectrometer failed to detect organic compounds on Mars, and both the Viking Labeled Release and the Viking Gas Exchange experiments indicated a reactive soil surface. These results have led to the widespread belief that there are oxidants in the martian soil. Since H2O2 is produced by photochemical processes in the atmosphere of Mars, and has been shown in the laboratory to reproduce closely the Viking LR results, it is a likely candidate for a martian soil oxidant. Here, we report on the results of a coupled soil/atmosphere transport model for H2O2 on Mars. Upon diffusing into the soil, its concentration is determined by the extent to which it is adsorbed and by the rate at which it is catalytically destroyed. An analytical model for calculating the distribution of H2O2 in the martian atmosphere and soil is developed. The concentration of H2O2 in the soil is shown to go to zero at a finite depth, a consequence of the nonlinear soil diffusion equation. The model is parameterized in terms of an unknown quantity, the lifetime of H2O2 against heterogeneous catalytic destruction in the soil. Calculated concentrations are compared with a H2O2 concentration of 30 nmoles/cm3, inferred from the Viking Labeled Release experiment. A significant result of this model is that for a wide range of H2O2 lifetimes (up to 10(5) years), the extinction depth was found to be less than 3 m. The maximum possible concentration in the top 4 cm is calculated to be approximately 240 nmoles/cm3, achieved with lifetimes of greater than 1000 years. Concentrations higher than 30 nmoles/cm3 require lifetimes of greater than 4.3 terrestrial years. For a wide range of H2O2 lifetimes, it was found that the atmospheric concentration is only weakly coupled with soil loss processes. Losses to the soil become significant only when lifetimes are less than a few hours. If there are depths below which H2O2 is not transported, it is plausible that organic compounds, protected from an oxidizing environment, may still exist. They would have been deposited by meteors, or be the organic remains of past life.

Atmosphere↗

Reflectance spectroscopy of ferric sulfate-bearing montmorillonites as Mars soil analog materials.

Spectroscopic analyses have shown that smectites enhanced in the laboratory with additional ferric species exhibit important similarities to those of the soils on Mars. Ferrihydrite in these chemically treated smectites has features in the visible to near-infrared region that resemble the energies and band strengths of features in reflectance spectra observed for several bright regions on Mars. New samples have been prepared with sulfate as well, because S was found by Viking to be a major component in the surface material on Mars. A suite of ferrihydrite-bearing and ferric sulfate-bearing montmorillonites, prepared with variable Fe3+ and S concentrations and variable pH conditions, has been analyzed using reflectance spectroscopy in the visible and infrared regions, Mössbauer spectroscopy at room temperature and 4 K, differential thermal analysis, and X-ray diffraction. These analyses support the formation of ferrihydrite of variable crystallinity in the ferrihydrite-bearing montmorillonites and a combination of schwertmannite and ferrihydrite in the ferric sulfate-bearing montmorillonites. Small quantities of poorly crystalline or nanophase forms of other ferric materials may also be present in these samples. The chemical formation conditions of the ferrihydrite-bearing and ferric sulfate-bearing montmorillonites influence the character of the low temperature Mössbauer sextets and the visible reflectance spectra. An absorption minimum is observed at 0.88-0.89 micrometers in spectra of the ferric sulfate-bearing samples, and at 0.89-0.92 micrometers in spectra of the ferrihydrate-bearing montmorillonites. Mössbauer spectra of the ferric sulfate-bearing montmorillonites indicate variable concentrations of ferrihydrite and schwertmannite in the interlaminar spaces and along grain surfaces. Dehydration under reduced atmospheric pressure conditions induces a greater effect on the adsorbed and interlayer water in ferrihydrite-bearing montmorillonite than on the water in ferric sulfate-bearing montmorillonite. Reflectance spectra of ferric sulfate-bearing montmorillonite include a strong 3-micrometers band that is more resistant to dry atmospheric conditions than the 3-micrometers band in spectra of similarly prepared ferrihydrite-bearing montmorillonites.

Aluminum Silicates↗

Production of low molecular weight hydrocarbons by volcanic eruptions on early Mars.

Methane and other larger hydrocarbons have been proposed as possible greenhouse gases on early Mars. In this work we explore if volcanic processes may have been a source for such molecules based on theoretical and experimental considerations. Geologic evidence and numerical simulations indicate that explosive volcanism was widely distributed throughout Mars. Volcanic lightning is typically produced in such explosive volcanism. Therefore this geologic setting was studied to determine if lightning could be a source for hydrocarbons in volcanic plumes. Volcanic lightning was simulated by focusing a high-energy infrared laser beam inside of a Pyrex reactor that contained the proposed volcanic gas mixture composed of 64% CH(4), 24% H(2), 10% H(2)O and 2% N(2), according to an accretion model and the nitrogen content measured in Martian meteorites. The analysis of products was performed by gas chromatography coupled to infrared and mass spectroscopy. Eleven hydrocarbons were identified among the products, of which acetylene (C(2)H(2)) was the most abundant. A thermochemical model was used to determine which hydrocarbons could arise only from volcanic heat. In this case, acetylene and ethylene are formed at magmatic temperatures. Our results indicate that explosive volcanism may have injected into the atmosphere of early Mars approximately 6 x 10(12) g yr(-1) of acetylene, and approximately 2 x 10(12) g yr(-1) of 1,3-butadiyne, both produced by volcanic lightning, approximately 5 x 10(11) g yr(-1) of ethylene produced by volcanic heat, and 10(13) g yr(-1) of methane.

Evolution, Chemical↗

Mars primordial crust: unique sites for investigating proto-biologic properties.

The Martian meteorite collection suggests that intact outcrops or boulder-scale fragments of the 4.5 Ga Martian crust exist within tens of meters of the present day surface of Mars. Mars may be the only planet where such primordial crust samples, representing the first 100 Ma of a planet's environment, are available. The primordial crust has been destroyed on Earth by plate tectonics and other geological phenomena and is buried on the Moon under hundreds or thousands of meters of megaregoltih. Early Mars appears to have been remarkably similar to early Earth, and samples of rock from the first few Ma or first 100 Ma may reveal "missing link" proto-biological forms that could shed light on the transition from abiotic organic chemistry to living cells. Such organic snapshots of nascent life are unlikely to be found on Earth.

Earth, Planet↗

Studies of biominerals relevant to the search for life on Mars.

The evidence of the water erosion on Mars is particularly interesting since present climatic conditions are such that liquid water cannot exist at the surface. But, if water was present on the planet in the past, there may have been life, too. Since the discovery of carbonates on Mars also may have very important implications on the possibility that life developed there, we are studying minerals that can have biotic or abiotic origin: calcite (CaCO(3)) and aragonite, a metastable state of calcite.We have analysed biomineral aragonite, in the form of recent sea shells, as well as crystals of mineral aragonite. Infrared spectroscopy in the 2-25 mum wavelength range reveals that, after thermal processing, the biotic samples have a different spectral behaviour from the abiotic ones. As a result, it is possible to distinguish abiotic mineral aragonite from aragonite of recent biological origin.Obviously, if life existed in the past on the Red Planet, we could expect to find "ancient" biotic carbonates, which should therefore be investigated, in order to search for a way of discriminating them from abiotic minerals. For this reason, at the beginning we have considered samples of crushed fossil shells of aragonite composition. Afterwards, in order to take into account that fossilization processes almost always produce a transformation of metastable form (aragonite) into more stable form (calcite), we also studied samples of mineral calcite and different types of fossils completely transformed into calcite. All these biotic fossil samples show the same spectral behaviour as the fresh biotic material after thermal annealing at 485 degrees C. Instead, the calcite behaves like abiotic aragonite.Furthermore, it is known that seashells and other biominerals are formed through an intimate association of inorganic materials with organic macromolecules. The macromolecules control the nucleation, structure, morphology, crystal orientation and spatial confinement of the inorganic phase: this differentiates biominerals from minerals. Analysing the aragonite or calcite fossils with a Scanning Electron Microscope, we found that the fossilization process did not modify the structure of the biominerals which maintain their microscopic characteristics. Looking at the morphology of fossil biominerals, it is evident that the crystals are arranged in complex architectures compared with the compact structure of the mineral crystals. In conclusion, the properties and structure of the biominerals are different from those of the minerals. The rapid increase of the crystalline structure developed under biotic conditions makes these minerals less resistant to thermal treatments, compared with samples of abiotic origin. This result holds both for recent shells as well as all fossil samples. The spectroscopic behaviour of all analysed calcium carbonates of biotic origin is different from that of the abiotic one. Therefore, the infrared spectroscopy is a valid technique to discern the origin of the samples and a powerful tool for analysing in-situ and "sample-return" Mars missions specimens. Also Optical and Scanning Electron Microscopy can be useful to support this type of studies.

Calcium Carbonate↗

A morphological view on potential niches for exobiology on Mars.

The discovery of microbiota in the Dry Valleys of Antarctica has encouraged the construction of new models of Martian ecosystems in order to determine if life could have once existed on Mars. The Antarctic cyanobacteria reside just below the surface of sandstone rocks where they are protected from the extreme cold and dry environment. Analogy with the Antarctic Dry Valleys supports speculation that hypothetical micro-organisms existed on Mars in the early history of the planet and could have migrated into suitable rocks as the availability of liquid water decreased. Although evidence for sandstone layers on Mars has not been substantiated, the palaeohydrology of Martian fluvial valleys (MFVs) reveals the evidence of lake bed sediment depositions which have formed consolidated sediments. As the MFVs formation may result from underground drainage processes, the sediment material would be expected to contain debris such as pumice washload, and pumilith of volcanic and meteoritic origin. These materials may have formed consolidated porous terrains similar to the Antarctic sandstone. Therefore, the endolithic model is consistent with the Martian liquid water habitat model of perenially ice-covered lakes.

Ecosystem↗

Oxygenic photosynthesis and the oxidation state of Mars.

The oxidation state of the Earth's surface is one of the most obvious indications of the effect of life on this planet. The surface of Mars is highly oxidized, as evidenced by its red color, but the connection to life is less apparent. Two possibilities can be considered. First, the oxidant may be photochemically produced in the atmosphere. In this case the fundamental source of O2 is the loss of H2 to space and the oxidant produced is H2O2. This oxidant would accumulate on the surface and thereby destroy any organic material and other reductants to some depth. Recent models suggest that diffusion limits this depth to a few meters. An alternative source of oxgyen is biological oxygen production followed by sequestration of organic material in sediments--as on the Earth. In this case, the net oxidation of the surface was determined billions of years ago when Mars was a more habitable planet and oxidative conditions could persist to great depths, over 100 m. Below this must be a compensating layer of biogenic organic material. Insight into the nature of past sources of oxidation on Mars will require searching for organics in the Martian subsurface and sediments.

Atmosphere↗

Peroxides and the survivability of microorganisms on the surface of Mars.

Results of the Viking mission seem to indicate that there is a ubiquitous layer of highly oxidizing aeolian material covering the Martian surface. This layer is thought to oxidize organic material that may settle on it, and is therefore responsible for the lack of detection of organic matter on the planet's surface by Viking. The mechanism that creates the oxidizing condition is not well understood, nor is the extent of the oxidation potential of this material. It has been suggested that the oxidizing nature of the soil is due to photochemical reactions which create hydrogen peroxide and superoxides in the surface soil. One question of importance to planetary protection regarding this material is, what is its potential for destroying terrestrial microorganisms, thus making the surface of Mars "self-sterilizing"? Using data obtained by the gas exchange experiment on Viking, and for simplicity assuming that all of the O2 released came from H2O2, the concentration range for H2O2 on the surface of Mars can be calculated to be 25-250 ppm. The microbial disinfection rate by H2O2 is concentration dependent, and is highly variable within the microbial community. Data from our laboratory indicate that certain soil bacteria survive and grow to stationary phase in 30,000 ppm H2O2. However, the total number of organisms decreases in the presence of H2O2. These results indicate that it is doubtful that the presence of H2O2 alone on Mars would make the surface "self-sterilizing".

Bacteria↗