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Responses across the gravity continuum: hypergravity to microgravity.

In response to hypergravity, it appears that the larger the animal, the greater the response, if present. Therefore, the response of a rat exceeds that of a mouse in the same hypergravity environment. When investigated in the microgravity environment of space flight, this appears to hold true. The lack of definitive data obtained in space for either species makes the extrapolation of the continuum to levels below Earth-gravity problematic. However, in systems where responses are detected for both space flight and acceleration by centrifugation, a gravitational continuum is present supporting the "principle of continuity". For those and similar systems, it appears that the use of hypergravity could be used to predict responses to space flight.

Animals↗

Integrating public perspectives in sample return planning.

Planning for extraterrestrial sample returns--whether from Mars or other solar system bodies--must be done in a way that integrates planetary protection concerns with the usual mission technical and scientific considerations. Understanding and addressing legitimate societal concerns about the possible risks of sample return will be a critical part of the public decision making process ahead. This paper presents the results of two studies, one with lay audiences, the other with expert microbiologists designed to gather information on attitudes and concerns about sample return risks and planetary protection. Focus group interviews with lay subjects, using generic information about Mars sample return and a preliminary environmental impact assessment, were designed to obtain an indication of how the factual content is perceived and understood by the public. A research survey of microbiologists gathered information on experts' views and attitudes about sample return, risk management approaches and space exploration risks. These findings, combined with earlier research results on risk perception, will be useful in identifying levels of concern and potential conflicts in understanding between experts and the public about sample return risks. The information will be helpful in guiding development of the environmental impact statement and also has applicability to proposals for sample return from other solar system bodies where scientific uncertainty about extraterrestrial life may persist at the time of mission planning.

Containment of Biohazards↗

Expression and affinity purification of recombinant proteins from plants.

With recent advances in plant biotechnology, transgenic plants have been targeted as an inexpensive means for the mass production of proteins for biopharmaceutical and industrial uses. However, the current plant purification techniques lack a generally applicable, economic, large-scale strategy. In this study, we demonstrate the purification of a model protein, beta-glucuronidase (GUS), by employing the protein calmodulin (CaM) as an affinity tag. In the proposed system, CaM is fused to GUS. In the presence of calcium, the calmodulin fusion protein binds specifically to a phenothiazine-modified surface of an affinity column. When calcium is removed with a complexing agent, e.g., EDTA, calmodulin undergoes a conformational change allowing the dissociation of the calmodulin-phenothiazine complex and, therefore, permitting the elution of the GUS-CaM fusion protein. The advantages of this approach are the fast, efficient, and economical isolation of the target protein under mild elution conditions, thus preserving the activity of the target protein. Two types of transformation methods were used in this study, namely, the Agrobacterium-mediated system and the viral-vector-mediated transformation system.

Blotting, Western↗

Earth analogs for Martian life. Microbes in evaporites, a new model system for life on Mars.

The prospect of life on Mars today is daunting. Especially problematic for a potential life form is a lack of water, particularly in a liquid state; extremely cold temperatures; ultraviolet and ionizing radiation; and soil oxidants. Yet, "oases" where life might persist have been suggested to occur in rocks (in analogy with endolithic microorganisms described from deserts around the world), in polar ice caps (in analogy with snow and ice algae) and in possible volcanic regions (in analogy with chemoautotrophs living in deep sea hydrothermal vents); all are critically examined. Microorganisms are known to be able to survive in salt crystals, and recently it has been shown that organisms can metabolize while encrusted in evaporites. Because evaporites are thought to occur on Mars and can attenuate light in the UV range while being far more transparent to radiation useful for photosynthesis (400-700 nm), and because of the properties of these "endoevaporitic" organisms, I propose that such communities provide a new model system for studying potential life on Mars. On the basis of this model, I suggest possibilities for site selection for future exobiological experiments on Mars.

Atmosphere↗

Post-Viking microbiology: new approaches, new data, new insights.

In the 20 years since the Viking experiments, major advances have been made in the areas of microbial systematics, microbial metabolism, microbial survival capacity, and the definition of environments on earth, suggesting that life is more versatile and tenacious than was previously appreciated. Almost all niches on earth which have available energy, and which are compatible with the chemistry of carbon-carbon bonds, are known to be inhabited by bacteria. The oldest known bacteria on earth apparently evolved soon after the formation of the planet, and are heat loving, hydrogen and/or sulfur metabolizing forms. Among the two microbial domains (kingdoms) is a great deal of metabolic diversity, with members of these forms being able to grow on almost any known energy source, organic or inorganic, and to utilize an impressive array of electron acceptors for anaerobic respiration. Both hydrothermal environments and the deep subsurface environments have been shown to support large populations of bacteria, growing on energy supplied by geothermal energy, thus isolating these ecosystems from the rest of the global biogeochemical cycles. This knowledge, coupled with new insights into the history of the solar system, allow one to speculate on possible evolution and survival of life forms on Mars.

Antarctic Regions↗

Regularity of daily life in relation to personality, age, gender, sleep quality and circadian rhythms.

A diary-like instrument to measure lifestyle regularity (the 'Social Rhythm Metric'-SRM) was given to 96 subjects (48 women, 48 men), 39 of whom repeated the study after at least one year, with additional objective measures of rest/activity. Lifestyle regularity as measured by the SRM related to age, morningness, subjective sleep quality and time-of-day variations in alertness, but not to gender, extroversion or neuroticism. Statistically significant test-retest correlations of about 0.4 emerged for SRM scores over the 12-30 month delay. Diary-based estimates of bedtime and waketime appeared fairly reliable. In a further study of healthy young men, 4 high SRM scorers ('regular') had a deeper nocturnal body temperature trough than 5 low SRM scorers ('irregular'), suggesting a better functioning circadian system in the 'regular' group.

Adult↗

Early and late mammalian responses to heavy charged particles.

This overview summarizes murine results on acute lethality responses, inactivation of marrow CFU-S and intestinal microcolonies, testes weight loss, life span shortening, and posterior lens opacification in mice irradiated with heavy charged particles. RBE-LET relationships for these mammalian responses are compared with results from in vitro studies. The trend is that the maximum RBE for in vivo responses tends to be lower and occurs at a lower LET than for inactivation of V79 and T-1 cells in culture. Based on inactivation cross sections, the response of CFU-S in vivo conforms to expectations from earlier studies with prokaryotic systems and mammalian cells in culture. Effects of heavy ions are compared with fission spectrum neutrons, and the results are consistent with the interpretation that RBEs are lower than for fission neutrons at about the same LET, probably due to differences in track structure. Issues discussed focus on challenges associated with assessments of early and late effects of charged particles based on dose, RBE and LET, and with the concordance or discordance of results obtained with in vivo and in vitro model systems. Models for radiation damage/repair and misrepair should consider effects observed with in vivo as well as in vitro model systems.

Animals↗

Human factors in spacecraft design.

This paper describes some of the salient implications of evolving mission parameters for spacecraft design. Among the requirements for future spacecraft are new, higher standards of living, increased support of human productivity, and greater accommodation of physical and cultural variability. Design issues include volumetric allowances, architecture and layouts, closed life support systems, health maintenance systems, recreational facilities, automation, privacy, and decor. An understanding of behavioral responses to design elements is a precondition for critical design decisions. Human factors research results must be taken into account early in the course of the design process.

Environment Design↗

Issues of exploration: human health and wellbeing during a mission to Mars.

Today, the tools are in our hands to enable us to travel away from our home planet and become citizens of the solar system. Even now, we are seriously beginning to develop the robust infrastructure that will make the 21st century the Century of Space Travel. But this bold step must be taken with due concern for the health, safety and wellbeing of future space explorers. Our long experience with space biomedical research convinces us that, if we are to deal effectively with the medical and biomedical issues of exploration, then dramatic and bold steps are also necessary in this field. We can no longer treat the human body as if it were composed of muscles, bones, heart and brain acting independently. Instead, we must lead the effort to develop a fully integrated view of the body, with all parts connected and fully interacting in a realistic way. This paper will present the status of current (2000) plans by the National Space Biomedical Research Institute to initiate research in this area of integrative physiology and medicine. Specifically, three example projects are discussed as potential stepping stones towards the ultimate goal of producing a digital human. These projects relate to developing a functional model of the human musculoskeletal system and the heart.

Adaptation, Physiological↗

Origins of species: acquired genomes and individuality.

Entire genomes with their accompanying protein synthetic systems are transferred throughout the biosphere primarily as bacteria and protists which become symbionts as they irreversibly integrate into pre-existing organisms to form more complex individuals. Individualization is stabilized by simultaneous transmission of once-separate heterologous genetic systems. The origin of new species is hypothesized to correlate with the acquisition, integration and subsequent inheritance of such acquired microbial genomes. These processes were recognized by Mereschkovsky ("Symbiogenesis" in Russian, 1909) and by Wallin ("Symbionticism", see p. 181, this issue).

Animals↗

Synthesis of the coenzymes adenosine diphosphate glucose, guanosine diphosphate glucose, and cytidine diphosphoethanolamine under primitive Earth conditions.

The nonenzymatic synthesis of the coenzymes adenosine diphosphate glucose (ADPG), guanosine diphosphate glucose (GDPG), and cytidine diphosphoethanolamine (CDP-ethanolamine) has been carried out under conditions considered to have been prevalent on the early Earth. The production of these compounds was performed by allowing simple precursor molecules to react under aqueous solutions, at moderate temperatures and short periods of time, with mediation by cyanamide or urea. These two condensing agents are considered to have been present in significant amounts on the primitive Earth and have been previously used in the nonenzymatic synthesis of several other important biochemical compounds. In our experiments, ADPG was obtained by heating glucose-1-phosphate (G1P) and ATP in the presence of cyanamide for 24 h at 70 degrees C. The reaction of G1P and GTP under the same conditions yielded GDPG. The cyanamide-mediated production of CDP-ethanolamine was carried out by reacting a mixture of ethanolamine phosphate and CTP for 24 h at 70 degrees C. The separation and identification of the reaction products was carried out by paper chromatography, thin-layer chromatography, high performance thin-layer chromatography, high performance liquid chromatography, both normal and reverse-phase, UV spectroscopy, enzymatic assays, and acid hydrolysis. Due to the mild conditions employed, and to the relative ease of these reactions, these studies offer a simple attractive system for the nonenzymatic synthesis of phosphorylated high-energy metabolic intermediates under conditions considered to have been prevalent on the ancient Earth.

Adenosine Diphosphate Glucose↗

Archaeal-eubacterial mergers in the origin of Eukarya: phylogenetic classification of life.

A symbiosis-based phylogeny leads to a consistent, useful classification system for all life. "Kingdoms" and "Domains" are replaced by biological names for the most inclusive taxa: Prokarya (bacteria) and Eukarya (symbiosis-derived nucleated organisms). The earliest Eukarya, anaerobic mastigotes, hypothetically originated from permanent whole-cell fusion between members of Archaea (e.g., Thermoplasma-like organisms) and of Eubacteria (e.g., Spirochaeta-like organisms). Molecular biology, life-history, and fossil record evidence support the reunification of bacteria as Prokarya while subdividing Eukarya into uniquely defined subtaxa: Protoctista, Animalia, Fungi, and Plantae.

Animals↗

Lunar habitat concept employing the space shuttle external tank.

The space shuttle external tank, which consists of a liquid oxygen tank, an intertank structure, and a liquid hydrogen tank, is an expendable structure used for approximately 8.5 min during each launch. A concept for outfitting the liquid oxygen tank-intertank unit for a 12-person lunar habitat is described. The concept utilizes existing structures and openings for both man and equipment access without compromising the structural integrity of the tank. Living quarters, instrumentation, environmental control and life support, thermal control, and propulsion systems are installed at Space Station Freedom. The unmanned habitat is then transported to low lunar orbit and autonomously soft landed on the lunar surface. Design studies indicate that this concept is feasible by the year 2000 with concurrent development of a space transfer vehicle and manned cargo lander for crew changeover and resupply.

Ecological Systems, Closed↗

The future of space medicine.

In November 2000, the National Aeronautics and Space Administration (NASA) and its partners in the International Space Station (ISS) ushered in a new era of space flight: permanent human presence in low-Earth orbit. As the culmination of the last four decades of human space flight activities. the ISS focuses our attention on what we have learned to date. and what still must be learned before we can embark on future exploration endeavors. Space medicine has been a primary part of our past success in human space flight, and will continue to play a critical role in future ventures. To prepare for the day when crews may leave low-Earth orbit for long-duration exploratory missions, space medicine practitioners must develop a thorough understanding of the effects of microgravity on the human body, as well as ways to limit or prevent them. In order to gain a complete understanding and create the tools and technologies needed to enable successful exploration. space medicine will become even more of a highly collaborative discipline. Future missions will require the partnership of physicians, biomedical scientists, engineers, and mission planners. This paper will examine the future of space medicine as it relates to human space exploration: what is necessary to keep a crew alive in space, how we do it today, how we will accomplish this in the future, and how the National Aeronautics and Space Administration (NASA) plans to achieve future goals.

Adaptation, Physiological↗

A novel reagentless sensing system for measuring glucose based on the galactose/glucose-binding protein.

The galactose/glucose-binding protein (GBP) is synthesized in the cytoplasm of Escherichia coli in a precursor form and exported into the periplasmic space upon cleavage of a 23-amino-acid leader sequence. GBP binds galactose and glucose in a highly specific manner. The ligand induces a hinge motion in GBP and the resultant protein conformational change constitutes the basis of the sensing system. The mglB gene, which codes for GBP, was isolated from the chromosome of E. coli using the polymerase chain reaction (PCR). Since wild-type GBP lacks cysteines in its structure, introducing this amino acid by site-directed mutagenesis ensures single-label attachment at specific sites with a sulfhydro-specific fluorescent probe. Site-directed mutagenesis by overlap extension PCR was performed to prepare three different mutants to introduce a single cysteine residue at positions 148, 152, and 182. Since these residues are not involved in ligand binding and since they are located at the edge of the binding cleft, they experience a significant change in environment upon binding of galactose or glucose. The sensing system strategy is based on the fluorescence changes of the probe as the protein undergoes a structural change on binding. In this work a reagentless sensing system has been rationally designed that can detect submicromolar concentrations of glucose. The calibration plots have a linear working range of three orders of magnitude. Although the system can sense galactose as well, this epimer is not a potential interfering substance since its concentration in blood is negligible.

Base Sequence↗

NASA' s life sciences and space radiation biology.

Plans for the various missions in which men and women are expected to participate during the next 10 years are outlined. Such missions include flights of up to three months duration in low earth orbit as well as possible short excursions to geosynchronous orbit. Research activities are described which cover the full spectrum of physiological and psychological responses to space flight. These activities are shown to contribute to the ongoing Shuttle program and the future Space Station. The paper includes a summary of the major technical thrusts needed to support extended habitation in space.

Adaptation, Physiological↗

Measurement of persistence in 1D diffusion.

Using a novel NMR scheme we observed persistence in 1D gas diffusion. Analytical approximations and numerical simulations have indicated that for an initially random array of spins undergoing diffusion, the probability p(t) that the average spin magnetization in a given region has not changed sign (i.e., "persists") up to time t follows a power law t(-straight theta), where straight theta depends on the dimensionality of the system. Using laser-polarized 129Xe gas, we prepared an initial "quasirandom" 1D array of spin magnetization and then monitored the ensemble's evolution due to diffusion using real-time NMR imaging. Our measurements are consistent with analytical and numerical predictions of straight theta approximately 0.12.

Diffusion↗

Multifractality in human heartbeat dynamics.

There is evidence that physiological signals under healthy conditions may have a fractal temporal structure. Here we investigate the possibility that time series generated by certain physiological control systems may be members of a special class of complex processes, termed multifractal, which require a large number of exponents to characterize their scaling properties. We report on evidence for multifractality in a biological dynamical system, the healthy human heartbeat, and show that the multifractal character and nonlinear properties of the healthy heart rate are encoded in the Fourier phases. We uncover a loss of multifractality for a life-threatening condition, congestive heart failure.

Adult↗