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At least 181 records · Page 10

High- and low-LET Radiation-induced Chromosome Aberrations in Human Epithelial Cells Cultured in 3-dimensional Matrices

Energetic heavy ions pose a great health risk to astronauts who participate in extended ISS missions and will be an even greater concern for future manned lunar and Mars missions. High-LET heavy ions are particularly effective in causing various biological effects, including cell inactivation, genetic mutations, cataracts and cancer induction. Most of these biological endpoints are closely related to chromosomal damage, which can be utilized as a biomarker for radiation insults. Previously, we had studied low- and high-LET radiation-induced chromosome aberrations in human epithelial cells cultured in 2-dimension (2D) using the multicolor banding fluorescence in situ hybridization (mBAND) technique. However, it has been realized that the biological response to radiation insult in a 2D in vitro cellular environment can differ significantly from the response in 3-dimension (3D) or at the actual tissue level. In this study, we cultured human epithelial cells in 3D to provide a more suitable model for human tissue. Human mammary epithelial cells (CH184B5F5/M10) were grown in Matrigel to form 3D structures, and exposed to Fe-ions at NASA Space Radiation Laboratory (NSRL) at the Brookhaven National Laboratory or 137Cs-gamma radiation source at the University of Texas MD Anderson Cancer Center. After exposure, cells were allowed to repair for 16hr before dissociation and subcultured at low density in 2D. G2 and metaphase chromosomes in the first cell cycle were collected in the first cell cycle after irradiation using a chemical-induced premature chromosome condensation (PCC) technique, and chromosome aberrations were analyzed using mBAND technique. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of interchromosomal aberrations (translocation to unpainted chromosomes) and intrachromosomal aberrations (inversions and deletions within a single painted chromosome). Our data indicate a significant difference in the chromosome aberration yield between 2D and 3D cell cultures after gamma exposures, but not after Fe ion exposures. Therefore, the Relative Biological Effect (RBE) for induction of chromosome aberrations obtained in a 2D model may not accurately represent RBE values obtained for tissue exposure.

Hada, M.↗

NEEMO 20: Science Training, Operations, and Tool Development

The 20th mission of the National Aeronautics and Space Administration (NASA) Extreme Environment Mission Operations (NEEMO) was a highly integrated evaluation of operational protocols and tools designed to enable future exploration beyond low-Earth orbit. NEEMO 20 was conducted from the Aquarius habitat off the coast of Key Largo, FL in July 2015. The habitat and its surroundings provide a convincing analog for space exploration. A crew of six (comprised of astronauts, engineers, and habitat technicians) lived and worked in and around the unique underwater laboratory over a mission duration of 14-days. Incorporated into NEEMO 20 was a diverse Science Team (ST) comprised of geoscientists from the Astromaterials Research and Exploration Science (ARES/XI) Division from the Johnson Space Center (JSC), as well as marine scientists from the Department of Biological Sciences at Florida International University (FIU). This team trained the crew on the science to be conducted, defined sampling techniques and operational procedures, and planned and coordinated the science focused Extra Vehicular Activities (EVAs). The primary science objectives of NEEMO 20 was to study planetary sampling techniques and tools in partial gravity environments under realistic mission communication time delays and operational pressures. To facilitate these objectives two types of science sites were employed 1) geoscience sites with available rocks and regolith for testing sampling procedures and tools and, 2) marine science sites dedicated to specific research focused on assessing the photosynthetic capability of corals and their genetic connectivity between deep and shallow reefs. These marine sites and associated research objectives included deployment of handheld instrumentation, context descriptions, imaging, and sampling; thus acted as a suitable proxy for planetary surface exploration activities. This abstract briefly summarizes the scientific training, scientific operations, and tool development conducted during NEEMO 20 with an emphasis on the primary lessons learned.

Graff, T.↗

The Hindlimb Unloading Rat Model: Literature Overview, Comparison with Spaceflight Data, and Technique Update

The hindlimb unloading (HU) rodent model is used extensively to study the response of many physiological systems to certain aspects of spaceflight, as well as to disuse and recovery from disuse for Earth benefits. This chapter describes the evolution of HU, and is divided into three sections. The first section examines the characteristics of 1063 articles using or reviewing the HU model, published between 1976 and April 1, 2004. The characteristics include number of publications, journals, countries, major physiological systems, method modifications, species, gender, genetic strains and ages of rodents, experiment duration, and countermeasures. The second section provides a comparison of results between space flown and Hu animals from the 14-day Cosmos 2044 mission. The final section describes modifications to HU required by different experimental paradigms and a method to protect the tail harness for long duration studies. HU in rodents has enabled improved understanding of the responses of the musculoskeletal, cardiovascular, immune, renal, neural, metabolic, and reproductive systems to unloading and/or to reloading on Earth with implications for both long-duration human spaceflight and disuse on Earth.

Morey-Holton, Emily↗

Design of Quiet Rotorcraft Approach Trajectories

A optimization procedure for identifying quiet rotorcraft approach trajectories is proposed and demonstrated. The procedure employs a multi-objective genetic algorithm in order to reduce noise and create approach paths that will be acceptable to pilots and passengers. The concept is demonstrated by application to two different helicopters. The optimized paths are compared with one another and to a standard 6-deg approach path. The two demonstration cases validate the optimization procedure but highlight the need for improved noise prediction techniques and for additional rotorcraft acoustic data sets.

Padula, Sharon L.↗

Oxygen isotopes in deep sea spherules

The determination of the genetic relationships between the dust and small particles in the solar system, and the meteorites and larger bodies are examined. Oxygen isotopes proved useful in the identification of such relationships between one meteorite group and another. Of the various samples of submillimeter extraterrestrial particles available for laboratory study, only the deep sea spherules are abundant enough for precise oxygen isotope analysis using existing techniques. Complications arise in interpretation of the isotopic data, since these particles were melted during passage through the Earth's atmosphere, and have been in contact with seawater for prolonged periods. Spherules that were originally silicates are considered with the originally metallic ones to deduce their preterrestrial isotopic compositions. The type 1 spherules which enter the atmosphere as metallic particles, contain only atmospheric oxygen. The type S spherules contain a mixture of atmospheric oxygen and their original extraterrestrial oxygen. It is suggested that the Earth's mesosphere is strongly enriched in heavy isotopes of oxygen at altitudes near 90 km at which the iron particles are oxidized. Fractionation due to the combined diffusion of O atoms and O2 molecules may be responsible.

Mayeda, T. K.↗

Systolic array IC for genetic computation

Measuring similarities between large sequences of genetic information is a formidable task requiring enormous amounts of computer time. Geneticists claim that nearly two months of CRAY-2 time are required to run a single comparison of the known database against the new bases that will be found this year, and more than a CRAY-2 year for next year's genetic discoveries, and so on. The DNA IC, designed at HP-ICBD in cooperation with the California Institute of Technology and the Jet Propulsion Laboratory, is being implemented in order to move the task of genetic comparison onto workstations and personal computers, while vastly improving performance. The chip is a systolic (pumped) array comprised of 16 processors, control logic, and global RAM, totaling 400,000 FETS. At 12 MHz, each chip performs 2.7 billion 16 bit operations per second. Using 35 of these chips in series on one PC board (performing nearly 100 billion operations per second), a sequence of 560 bases can be compared against the eventual total genome of 3 billion bases, in minutes--on a personal computer. While the designed purpose of the DNA chip is for genetic research, other disciplines requiring similarity measurements between strings of 7 bit encoded data could make use of this chip as well. Cryptography and speech recognition are two examples. A mix of full custom design and standard cells, in CMOS34, were used to achieve these goals. Innovative test methods were developed to enhance controllability and observability in the array. This paper describes these techniques as well as the chip's functionality. This chip was designed in the 1989-90 timeframe.

Anderson, D.↗

Molecular Technique to Understand Deep Microbial Diversity

Current sequencing-based and DNA microarray techniques to study microbial diversity are based on an initial PCR (polymerase chain reaction) amplification step. However, a number of factors are known to bias PCR amplification and jeopardize the true representation of bacterial diversity. PCR amplification of the minor template appears to be suppressed by the exponential amplification of the more abundant template. It is widely acknowledged among environmental molecular microbiologists that genetic biosignatures identified from an environment only represent the most dominant populations. The technological bottleneck has overlooked the presence of the less abundant minority population, and underestimated their role in the ecosystem maintenance. To generate PCR amplicons for subsequent diversity analysis, bacterial l6S rRNA genes are amplified by PCR using universal primers. Two distinct PCR regimes are employed in parallel: one using normal and the other using biotinlabeled universal primers. PCR products obtained with biotin-labeled primers are mixed with streptavidin-labeled magnetic beads and selectively captured in the presence of a magnetic field. Less-abundant DNA templates that fail to amplify in this first round of PCR amplification are subjected to a second round of PCR using normal universal primers. These PCR products are then subjected to downstream diversity analyses such as conventional cloning and sequencing. A second round of PCR amplified the minority population and completed the deep diversity picture of the environmental sample.

Vaishampayan, Parag A.↗

On the Use of Optimization Techniques for Turbulence Model Calibration

This article discusses the use of numerical optimization procedures to aid in the calibration of turbulence model coefficients. Such methods would increase the rigor and repeatability of the calibration procedure by requiring clearly defined and objective optimization metrics, and could be used to identify unique combinations of coefficient values for specific flow problems. The approach is applied to the re-calibration of an explicit algebraic Reynolds stress model for the incompressible planar mixing layer using the Nelder-Mead simplex algorithm and a micro-genetic algorithm with minimally imposed constraints. Three composite fitness functions, each based upon the error in the mixing layer growth rate and the normal and shear components of the Reynolds stresses, are investigated. The results demonstrate a significant improvement in the target objectives through the adjustment of three pressure-strain coefficients. Adjustments of additional coefficients provide little further benefit. Issues regarding the effectiveness of the fitness functions and the efficiency of the optimization algorithms are also discussed.

Yoder, Dennis A.↗

Relative effectiveness of HZE iron-56 particles for the induction of cytogenetic damage in vivo

One of the risks of prolonged manned space flight is the exposure of astronauts to radiation from galactic cosmic rays, which contain heavy ions such as (56)Fe. To study the effects of such exposures, experiments were conducted at the Brookhaven National Laboratory by exposing Wistar rats to high-mass, high-Z, high-energy (HZE) particles using the Alternating Gradient Synchrotron (AGS). The biological effectiveness of (56)Fe ions (1000 MeV/nucleon) relative to low-LET gamma rays and high-LET alpha particles for the induction of chromosome damage and micronuclei was determined. The mitotic index and the frequency of chromosome aberrations were evaluated in bone marrow cells, and the frequency of micronuclei was measured in cells isolated from the trachea and the deep lung. A marked delay in the entry of cells into mitosis was induced in the bone marrow cells that decreased as a function of time after the exposure. The frequencies of chromatid aberrations and micronuclei increased as linear functions of dose. The frequency of chromosome aberrations induced by HZE particles was about 3.2 times higher than that observed after exposure to (60)Co gamma rays. The frequency of micronuclei in rat lung fibroblasts, lung epithelial cells, and tracheal epithelial cells increased linearly, with slopes of 7 x 10(-4), 12 x 10(-4), and 11 x 10(-4) micronuclei/binucleated cell cGy(-1), respectively. When genetic damage induced by radiation from (56)Fe ions was compared to that from exposure to (60)Co gamma rays, (56)Fe-ion radiation was between 0.9 and 3.3 times more effective than (60)Co gamma rays. However, the HZE-particle exposures were only 10-20% as effective as radon in producing micronuclei in either deep lung or tracheal epithelial cells. Using microdosimetric techniques, we estimated that 32 cells were hit by delta rays for each cell that was traversed by the primary HZE (56)Fe particle. These calculations and the observed low relative effectiveness of the exposure to HZE particles suggest that at least part of the cytogenetic damage measured was caused by the delta rays. Much of the energy deposited by the primary HZE particles may result in cell killing and may therefore be "wasted" as far as production of detectable micronuclei is concerned. The role of wasted energy in studies of cancer induction may be important in risk estimates for exposure to HZE particles.

Non-NASA Center↗

M-Band Analysis of Chromosome Aberrations in Human Epithelial Cells Induced By Low- and High-Let Radiations

Energetic primary and secondary particles pose a health risk to astronauts in extended ISS and future Lunar and Mars missions. High-LET radiation is much more effective than low-LET radiation in the induction of various biological effects, including cell inactivation, genetic mutations, cataracts and cancer. Most of these biological endpoints are closely correlated to chromosomal damage, which can be utilized as a biomarker for radiation insult. In this study, human epithelial cells were exposed in vitro to gamma rays, 1 GeV/nucleon Fe ions and secondary neutrons whose spectrum is similar to that measured inside the Space Station. Chromosomes were condensed using a premature chromosome condensation technique and chromosome aberrations were analyzed with the multi-color banding (mBAND) technique. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of both interchromosomal (translocation to unpainted chromosomes) and intrachromosomal aberrations (inversions and deletions within a single painted chromosome). Results of the study confirmed the observation of higher incidence of inversions for high-LET irradiation. However, detailed analysis of the inversion type revealed that all of the three radiation types in the study induced a low incidence of simple inversions. Half of the inversions observed in the low-LET irradiated samples were accompanied by other types of intrachromosome aberrations, but few inversions were accompanied by interchromosome aberrations. In contrast, Fe ions induced a significant fraction of inversions that involved complex rearrangements of both the inter- and intrachromosome exchanges.

Hada, M.↗

The hindlimb unloading rat model: literature overview, technique update and comparison with space flight data

The hindlimb unloading rodent model is used extensively to study the response of many physiological systems to certain aspects of space flight, as well as to disuse and recovery from disuse for Earth benefits. This chapter describes the evolution of hindlimb unloading, and is divided into three sections. The first section examines the characteristics of 1064 articles using or reviewing the hindlimb unloading model, published between 1976 and April 1, 2004. The characteristics include number of publications, journals, countries, major physiological systems, method modifications, species, gender, genetic strains and ages of rodents, experiment duration, and countermeasures. The second section provides a comparison of results between space flown and hindlimb unloading animals from the 14-day Cosmos 2044 mission. The final section describes modifications to hindlimb unloading required by different experimental paradigms and a method to protect the tail harness for long duration studies. Hindlimb unloading in rodents has enabled improved understanding of the responses of the musculoskeletal, cardiovascular, immune, renal, neural, metabolic, and reproductive systems to unloading and/or to reloading on Earth with implications for both long-duration human space flight and disuse on Earth.

Flight Experiment↗

Models and molecular approaches to assessing the effects of the microgravity environment on vertebrate development

The extent to which gravity, and especially the lack thereof, can affect normal development in higher organisms is poorly understood. Underlying this question is the assumption that normal development depends on the embryo's ability to maintain a programmed temporal and spatial coordination of morphogenetic events. There are several reports documenting the apparently normal development of several vertebrate species, including mammals, under conditions of exposure to space flight during various periods of the development process. Evidence to the contrary also exists and it is therefore likely that some alterations in morphology do occur in a microgravity environment. Although subsequent development may appear overtly normal, more subtle abnormalities result. In all studies, the evaluation is restricted by the few numbers of specimens that can be examined and the relatively insensitive techniques for assessing potentially subtle effects. In the present discussion, we summarize some observations of mammalian development made in microgravity and consider which stages might be expected to be differentially sensitive to altered gravity conditions. While we emphasize mammalian development, we discuss the suitability of another model system for examining such effects in a cross-species context. Furthermore, we consider recent developments in our understanding of the molecular genetic program regulating embryogenesis that could serve as markers for assessing perturbations of development.

Cosmos 1129 Project↗

Cardiovascular tissues contain independent circadian clocks

Acute cardiovascular events exhibit a circadian rhythm in the frequency of occurrence. The mechanisms underlying these phenomena are not yet fully understood, but they may be due to rhythmicity inherent in the cardiovascular system. We have begun to characterize rhythmicity of the clock gene mPer1 in the rat cardiovascular system. Luciferase activity driven by the mPer1 gene promoter is rhythmic in vitro in heart tissue explants and a wide variety of veins and arteries cultured from the transgenic Per1-luc rat. The tissues showed between 3 and 12 circadian cycles of gene expression in vitro before damping. Whereas peak per1-driven bioluminescence consistently occurred during the late night in the heart and all arteries sampled, the phases of the rhythms in veins varied significantly by anatomical location. Varying the time of the culture procedure relative to the donor animal's light:dark cycle revealed that, unlike some other rat tissues such as liver, the phases of in vitro rhythms of arteries, veins, and heart explants were affected by culture time. However, phase relationships among tissues were consistent across culture times; this suggests diversity in circadian regulation among components of the cardiovascular system.

Non-NASA Center↗

Implementation of Combinatorial Optimization Techniques for Automated Fiber Placement Through Thickness Defect Stack-Up Minimization

The Computer Aided Process Planning (CAPP) module was developed to facilitate and accelerate the process planning workflow for Automated Fiber Placement (AFP). CAPP assists process planners in identifying optimal starting point locations and layup strategies for each ply of a laminate. Ply optimization operates on measurement and scoring of geometry-based defects such as gaps, overlaps, angle deviation, and steering. This paper expands on the established framework for analyzing defect stack-up through thickness of a laminate. Four different combinatorial optimization algorithms are implemented and evaluated: (1) genetic algorithm, (2) differential evolution, (3) particle swarm, and (4) greedy search. The algorithms identify the optimal combination of ply-level layup strategies, by scoring potential laminates on defect stacking, using two different objective functions. A final optimization approach is also presented which trades some performance for a large gain in efficiency. These approaches are compared to a randomized combination using a complex tool surface in a virtual case study. The result is a streamlined methodology for comparing different laminate-level manufacturing strategies and minimizing the through thickness defect stack up.

CAPP↗

A method to identify and characterize Z-DNA binding proteins using a linear oligodeoxynucleotide

An oligodeoxynucleotide that readily flips to the Z-DNA conformation in 10mM MgCl2 was produced by using Klenow enzyme to incorporate 5-bromodeoxycytosine and deoxyguanosine into a (dC-dG)22 template. During synthesis the oligomer can be labeled with 32P to high specific activity. The labeled oligodeoxynucleotide can be used in bandshift experiment to detect proteins that bind Z-DNA. This allows the binding specificity of such proteins to be determined with high reliability using unlabeled linear and supercoiled DNA competitors. In addition, because the radioactive oligodeoxynucleotide contains bromine atoms, DNA-protein complexes can be readily crosslinked using UV light. This allows an estimate to be made of the molecular weight of the proteins that bind to the radioactive probe. Both techniques are demonstrated using a goat polyclonal anti-Z-DNA antiserum.

NASA Program Exobiology↗

An Invited Preface for the Following Book: Astrobiologia, Uma Ciencia Emergente

Since the dawn of civilization, we have beheld at the beauty and wonder of the natural world around us and wondered how it came to be. We have pondered the past, and have been intrigued about the future. For this we are unique. Our ancestors looked to the vastness of space and thought surely there are others out there. We are now at a new time in human history where we can address these age-old questions with a scientific approach and study rigorously the three big questions of astrobiology: Where do we come from? Where are we going? and Are we alone? These fundamental questions of astrobiology correspond to those of humanity, and arguably, what makes us human. And so we cannot help but be drawn to the field. Unlike other scientific disciplines, Astrobiology draws on the latest advances in a multitude of fields, from evolutionary and molecular biology, to prebiotic and interstellar chemistry, from astrophysics to astronomy, with a healthy dose of earth and planetary science. Astrobiology is in reality a "metadiscipline" drawing on useful science wherever it is to be found. From a practical point of view, this endeavor requires the interaction of scientists who might not normally meet each other, much less work on a common research project. And, unlike most other scientific disciplines, Astrobiology has implications for how we see ourselves, and how we interact with the earth and beyond. "Where do we come from" touches on the "why" questions that have intrigued not just scientists but philosophers and theologians. "Where are we going" adds to these an economic and political involvement that is currently being played out with discussions of climate change. "And are we alone" will someday force us to face the fact that we as living creatures are not unique, or perhaps that we are utterly alone in the universe, the result of a chemical history that was so improbable as to result in a sample size of one. Either result will force ethical considerations of either "the other" and their relationship to us, or our solitude and thus responsibility as the only life form in our cosmos. So what is Astrobiology? Let's start with the "Where do we come from?" A biologist will approach this looking at the evolution of life on earth, using such traditional tools as comparative anatomy and paleontology and newer tools such as molecular techniques. But this doesn't address why this happened the way it did without a comprehensive understanding of the environment. What was the temperature at such and such a time? Was the earth in a snowball phase or being bombarded by meteorites -- or even just a single large, well-placed one such as struck off the Yucatan peninsula 65 million years ago. This event could not have been predicted by population genetics alone, yet it had the most profound influence on our evolution as without it, we could still be in a world dominated by dinosaurs with the mammals cowering under cover. But it is not enough to go back to LUCA, the Last Universal Common Ancestor of all extant life. One must go back to the dawn of life. How did life arise? What was the environmental backdrop that allowed it to happen? How did we happen to end up with a habitable planet? Indeed, what is the origin and evolution of our solar system, galaxy, biogenic elements all the way back to the Big Bang. The "Where are we going?" tends to be ignored in many astrobiology programs, but in fact this is of the most immediate importance to us. Whereas the past was dominated by physical and chemical processes, and organismal interactions, the future has a new major player: us. While we probably don't have the power (yet) to stop our galaxy passing through another, or even reset the sun or stop the movement of the moon away from the earth, all of these things will influence the future of life on earth. We are already proving that we have the power to visit other bodies in our solar system, either with humans or our robotic surrogates. We are changing our atmospheric composition and thus our climate. We have the power to render species extinct, including our own. But we also have the power to use these tools for the common good, to extend our lifetimes and reclaim our rivers and forests. Which will it be? And then there is the question where science fiction becomes a reality: "Are we alone?" While many people are anxious to find signs of intelligent life out there, such a creature may not share either our curiosity or values. But what if there was a beneficent alien civilization that could communicate with us, perhaps forging a mutual understanding? More likely in the forseeable future is finding a small life, less evolved, life form. Note I did not use the word "simple" as there is nothing "simple" about life, ever. Which brings us back to the question: what is life? So where does Brazil come in? For nearly a decade there has been interest in a Brazilian astrobiology program, from a small side meeting held by the Brazilian Exobiology Program (BEP) of the Brazilian National Research Council, held in Rio de Janeiro, Brazil, on August 12, 2009, in association with the IAU Assembly to subsequent workshops held in several locations and membership of the Brazilian program in the NASA Astrobiology Institute (NAI). Each time I go to Brazil, I am impressed by the enthusiasm of the community, both scientific and student, the latter an excellent omen for the future of astrobiology in Brazil. Facilities are being built to supplement the natural laboratories that Brazil is blessed to have. In my own lab I have been privileged to have a wonderful Brazilian postdoctoral fellow, Dr. Ivan Paulino-Lima, resulting in a daily reminder of the program. I am honored to have been part of the development of astrobiology in Brazil, and hope that this relationship will continue to flourish. Clearly to advance astrobiology needs new knowledge, a reorganization of that which is known, and space missions. To help the reader join on this quest, what follows is a buffet of topics that will allow the reader to nibble on the richness that is Astrobiology. And, like a fabulous meal, it should make you want more. Welcome to Astrobiology.

Are we alone?↗

Biology and the Exploration of Mars

Until recent years the origin of life and its possible occurrence elsewhere in the universe have been matters for speculation only. The rapid growth of molecular biology since 1940 has, to be sure, made it possible to discuss life's origins in far more precise and explicit terms than was possible earlier; and the subject entered a new experimental phase in the 1950's with successful abiogenic synthesis of important biochemical substances in conditions simulating the presumptive environment of the primitive Earth. But the real transformation that the subject has undergone stems from the spectacular growth of space technology in the last decade. The possibility of life's origin and occurrence on planets other than ours is no longer limited to idle speculation: it has entered the realm of the testable, of science in the strict sense. Given the rockets now available, and especially those available by 1969, it has become fully realistic to consider plans for the biological exploration of Mars. The study that this report seeks to interpret was initiated in June, 1964, by the Space Science Board of the National Academy of Sciences to examine this possibility. The working group comprised 36 people representing a broad spectrum of scientific interests: evolutionary biology, genetics, microbiology, biochemistry and molecular biology, animal physiology, soil chemistry, organic chemistry, planetary astronomy, geochemistry, and theoretical physics. The participants included some with considerable prior involvement in problems of space exploration and others with none. Advice was also sought outside the group of immediate participants on the potentialities of selected analytical methods for the experimental study of extraterrestrial life and its environment. More than 30 individuals contributed in this fashion written assessments of techniques in which they were particularly well versed. Our task was to examine the scientific foundations and merits of the proposal to undertake a biological exploration of Mars. What are the potential scientific yields? How valuable, if attained, would they be? What, in fact, is the possibility of life occurring on Mars? And of our detecting it with available and foreseeable technology? What could be achieved by further astronomical work from Earth? by Martian fly-by missions? by Martian orbiters? and Martian Landers? What payloads would we recommend for planetary missions? What timing and over-all strategy would we recommend for Martian exploration were we to consider it worthwhile at all? In brief, the over-all purpose was to recommend to. the government, through the Academy's Space Science Board, whether or not a biological exploration of Mars should be included in the nation's space program over the next few decades; and, further, to outline what that program, if any, should be.

CONFERENCE↗

Evolution of catalytic RNA in the laboratory

We are interested in the biochemistry of existing RNA enzymes and in the development of RNA enzymes with novel catalytic function. The focal point of our research program has been the design and operation of a laboratory system for the controlled evolution of catalytic RNA. This system serves as working model of RNA-based life and can be used to explore the catalytic potential of RNA. Evolution requires the integration of three chemical processes: amplification, mutation, and selection. Amplification results in additional copies of the genetic material. Mutation operates at the level of genotype to introduce variability, this variability in turn being expressed as a range of phenotypes. Selection operates at the level of phenotype to reduce variability by excluding those individuals that do not conform to the prevailing fitness criteria. These three processes must be linked so that only the selected individuals are amplified, subject to mutational error, to produce a progeny distribution of mutant individuals. We devised techniques for the amplification, mutation, and selection of catalytic RNA, all of which can be performed rapidly in vitro within a single reaction vessel. We integrated these techniques in such a way that they can be performed iteratively and routinely. This allowed us to conduct evolution experiments in response to artificially-imposed selection constraints. Our objective was to develop novel RNA enzymes by altering the selection constraints in a controlled manner. In this way we were able to expand the catalytic repertoire of RNA. Our long-range objective is to develop an RNA enzyme with RNA replicase activity. If such an enzyme had the ability to produce additional copies of itself, then RNA evolution would operate autonomously and the origin of life will have been realized in the laboratory.

Joyce, Gerald F.↗