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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 145 records · Page 8
All sourcesNASA NTRSDOE OSTI
NASA NTRS2010

Miniature Bioreactor System for Long-Term Cell Culture

A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.

Gonda, Steve R.↗
NASA NTRS2010

Utilization of Alternate Chirality Enantiomers in Microbial Communities

Our previous study of chirality led to interesting findings for some anaerobic extremophiles: the ability to metabolize substrates with alternate chirality enantiomers of amino acids and sugars. We have subsequently found that not just separate microbial species or strains but entire microbial communities have this ability. The functional division within a microbial community on proteo- and sugarlytic links was also reflected in a microbial diet with L-sugars and D-amino acids. Several questions are addressed in this paper. Why and when was this feature developed in a microbial world? Was it a secondary de novo adaptation in a bacterial world? Or is this a piece of genetic information that has been left in modern genomes as an atavism? Is it limited exclusively to prokaryotes, or does this ability also occur in eukaryotes? In this article, we have used a broader approach to study this phenomenon using anaerobic extremophilic strains from our laboratory collection. A series of experiments were performed on physiologically different groups of extremophilic anaerobes (pure and enrichment cultures). The following characteristics were studied: 1) the ability to grow on alternate chirality enantiomers -- L-sugars and D- amino acids; 2) Growth-inhibitory effect of alternate chirality enantiomers; 3) Stickland reaction with alternate chirality amino acids. The results of this research are presented in this paper.

Pikuta, Elena V.↗
NASA NTRS2011

Low Fluid Shear Culture of Staphylococcus Aureus Represses hfq Expression and Induces an Attachment-Independent Biofilm Phenotype

Background: The opportunistic pathogen, Staphylococcus aureus, experiences fluctuations in fluid shear during infection and colonization of a human host. Colonization frequently occurs at mucus membrane sites such as in the gastrointestinal tract where the bacterium may experience low levels of fluid shear. The response of S. aureus to low fluid shear remains unclear. Methods: S. aureus was cultured to stationary phase using Rotating-Wall Vessel (RWV) bioreactors which produce a physiologically relevant low fluid shear environment. The bacterial aggregates that developed in the RWV were evaluated by electron microscopy as well as for antibiotic resistance and other virulence-associated stressors. Genetic expression profiles for the low-shear cultured S. aureus were determined by microarray analysis and quantitative real-time PCR. Results: Planktonic S. aureus cultures in the low-shear environment formed aggregates completely encased in high amounts of extracellular polymeric substances. In addition, these aggregates demonstrated increased antibiotic resistance indicating attachment-independent biofilm formation. Carotenoid production in the low-shear cultured S. aureus was significantly decreased, and these cultures displayed an increased susceptibility to oxidative stress and killing by whole blood. The hfq gene, associated with low-shear growth in Gram negative organisms, was also found to be down-regulated in S. aureus. Conclusions: Collectively, this data suggests that S. aureus decreases virulence characteristics in favor of a biofilm-dwelling colonization phenotype in response to a low fluid shear environment. Furthermore, the identification of an Hfq response to low-shear culture in S. aureus, in addition to the previously reported responses in Gram negative organisms, strongly suggests an evolutionarily conserved response to mechanical stimuli among structurally diverse prokaryotes.

Ott, C. Mark↗
NASA NTRS2011

Extremophile Diatoms: Implications to the Drake Equation

Diatoms are unicellular Eukaryotes that (as a group and phylogenetically) are not strictly regarded as extremophiles , since the vast majority of diatoms are mesophilic photoautotrophs. However, among the terrestrial Eukaryotes, diatoms are by far the single group of organisms with the ability to inhabit the greatest range of hostile environments on Earth. They are the dominant eukaryotes in the polar regions; in fumaroles, hot springs and geysers; and in hypersaline and hyperalkaline lakes and pools. Cryophilic species such as Fragilaria sublinearis and Chaetoceras fragilis are able to carry out respiration at extremely low rates at low temperatures in darkness. The Drake Equation refers to the likelihood of there being intelligent life at the technological level of electromagnetic communication. However, consideration of the range of conditions suitable for the habitability of eukaryotic diatoms and prokaryotic extremophiles, the likelihood that life exists elsewhere in the cosmos becomes many orders of magnitude greater than that predicted by the classical Drake Equation. In this paper we review the characteristics of diatoms as eukaryotic extremophiles and consider the implications to adjustments needed to the Drake Equation to assess the possibility that life exists elsewhere in the Universe.

Sterrenburg, Frithjof A. S.↗
NASA NTRS2011

Comets as Parent Bodies of CI1 Carbonaceous Meteorites and Possible Habitats of Ice-Microbiota

Recent studies of comets and cometary dust have confirmed the presence of biologically relevant organic molecules along with clay minerals and water ice. It is also now well established by deuterium/hydrogen ratios that the CI1 carbonaceous meteorites contain indigenous extraterrestrial water. The evidence of extensive aqueous alteration of the minerals in these meteorites led to the hypothesis that water-bearing asteroids or comets represent the parent bodies of the CI1 (and perhaps CM2) carbonaceous meteorites. These meteorites have also been shown to possess a diverse array of complex organics and chiral and morphological biomarkers. Stable isotope studies by numerous independent investigators have conclusively established that the complex organics found in these meteorites are both indigenous and extraterrestrial in nature. Although the origin of these organics is still unknown, some researchers have suggested that they originated by unknown abiotic mechanisms and may have played a role in the delivery of chiral biomolecules and the origin of life on Early Earth. In this paper we review these results and investigate the thermal history of comets. We show that permanent as well as transient domains of liquid water can be maintained on a comet under a plausible set of assumptions. With each perihelion passage of a comet volatiles are preferentially released, and during millions of such passages the comet could shed crustal debris that may survive transit through the Earth s atmosphere as a carbonaceous meteorite. We review the current state of knowledge of comets and carbonaceous meteorites. We also present the results of recent studies on the long-term viability of terrestrial ice-microbiota encased in ancient glacial ice and permafrost. We suggest that the conditions which have been observed to prevail on many comets do not preclude either survivability (or even the active metabolism and growth) of many types of eukaryotic and prokaryotic microbial extremophiles-including algae, cyanobacteria, bacteria and archaea. It is argued that the chemical and morphological biomarkers detected on comets and carbonaceous meteorites can be explained by ancient microbial activity without the need to invoke unknown abiotic production mechanisms.

Wickramasinghe, N. Chandra↗
NASA NTRS2012

Purifying, Separating, and Concentrating Cells From a Sample Low in Biomass

Frequently there is an inability to process and analyze samples of low biomass due to limiting amounts of relevant biomaterial in the sample. Furthermore, molecular biological protocols geared towards increasing the density of recovered cells and biomolecules of interest, by their very nature, also concentrate unwanted inhibitory humic acids and other particulates that have an adversarial effect on downstream analysis. A novel and robust fluorescence-activated cell-sorting (FACS)-based technology has been developed for purifying (removing cells from sampling matrices), separating (based on size, density, morphology), and concentrating cells (spores, prokaryotic, eukaryotic) from a sample low in biomass. The technology capitalizes on fluorescent cell-sorting technologies to purify and concentrate bacterial cells from a low-biomass, high-volume sample. Over the past decade, cell-sorting detection systems have undergone enhancements and increased sensitivity, making bacterial cell sorting a feasible concept. Although there are many unknown limitations with regard to the applicability of this technology to environmental samples (smaller cells, few cells, mixed populations), dogmatic principles support the theoretical effectiveness of this technique upon thorough testing and proper optimization. Furthermore, the pilot study from which this report is based proved effective and demonstrated this technology capable of sorting and concentrating bacterial endospore and bacterial cells of varying size and morphology. Two commercial off-the-shelf bacterial counting kits were used to optimize a bacterial stain/dye FACS protocol. A LIVE/DEAD BacLight Viability and Counting Kit was used to distinguish between the live and dead cells. A Bacterial Counting Kit comprising SYTO BC (mixture of SYTO dyes) was employed as a broad-spectrum bacterial counting agent. Optimization using epifluorescence microscopy was performed with these two dye/stains. This refined protocol was further validated using varying ratios and mixtures of cells to ensure homogenous staining compared to that of individual cells, and were utilized for flow analyzer and FACS labeling. This technology focuses on the purification and concentration of cells from low-biomass spacecraft assembly facility samples. Currently, purification and concentration of low-biomass samples plague planetary protection downstream analyses. Having a capability to use flow cytometry to concentrate cells out of low-biomass, high-volume spacecraft/ facility sample extracts will be of extreme benefit to the fields of planetary protection and astrobiology. Successful research and development of this novel methodology will significantly increase the knowledge base for designing more effective cleaning protocols, and ultimately lead to a more empirical and true account of the microbial diversity present on spacecraft surfaces. Refined cleaning and an enhanced ability to resolve microbial diversity may decrease the overall cost of spacecraft assembly and/or provide a means to begin to assess challenging planetary protection missions.

Benardini, James N.↗
NASA NTRS2012

Fluorescence-Activated Cell Sorting of Live Versus Dead Bacterial Cells and Spores

This innovation is a coupled fluorescence-activated cell sorting (FACS) and fluorescent staining technology for purifying (removing cells from sampling matrices), separating (based on size, density, morphology, and live versus dead), and concentrating cells (spores, prokaryotic, eukaryotic) from an environmental sample.

Bernardini, James N.↗
NASA NTRS2010

Metagenomic Study of Iron Homeostasis in Iron Depositing Hot Spring Cyanobacterial Community

Introduction: It is not clear how an iron-rich thermal hydrosphere could be hospitable to cyanobacteria, since reduced iron appears to stimulate oxidative stress in all domains of life and particularly in oxygenic phototrophs. Therefore, metagenomic study of cyanobacterial community in iron-depositing hot springs may help elucidate how oxygenic prokaryotes can withstand the extremely high concentrations of reactive oxygen species (ROS) produced by interaction between environmental Fe2+ and O2. Method: Anchor proteins from various species of cyanobacteria and some anoxygenic phototrophs were selected on the basis of their hypothetical role in Fe homeostasis and the suppression of oxidative stress and were BLASTed against the metagenomes of iron-depositing Chocolate Pots and freshwater Mushroom hot springs. Results: BLASTing proteins hypothesized to be involved in Fe homeostasis against the microbiomes from the two springs revealed that iron-depositing hot spring has a greater abundance of defensive proteins such as bacterioferritin comigratory protein (Bcp) and DNA-binding Ferritin like protein (Dps) than a fresh-water hot spring. One may speculate that the abundance of Bcp and Dps in an iron-depositing hot spring is connected to the need to suppress oxidative stress in bacteria inhabiting environments with high Fe2+ concnetration. In both springs, Bcp and Dps are concentrated within the cyanobacterial fractions of the microbial community (regardless of abundance). Fe3+ siderophore transport (from the transport system permease protein query) may be less essential to the microbial community of CP because of the high [Fe]. Conclusion: Further research is needed to confirm that these proteins are unique to photoautotrophs such as those living in iron-depositing hot spring.

Brown, I.↗
NASA NTRS2014

The Present Habitability Potential of Gale Crater: What We Have Learned So Far From Mars Science Laboratory

The Mars Science Laboratory mission has comprehensively interrogated the surface environment of Mars as it explores Gale Crater. Both chemical and physical attributes of the present environment have been measured over the course of the mission, enabling us to compare the present state of the martian surface with the environmental requirements of prokaryotic microbes. While this approach does not exclude the possibility of martian life that may have evolved to adapt to the present conditions, it is advantageous in that it allows us to evaluate environmental requirements of known life and also provide insight into the likelihood of forward contamination by Earth organisms with the comparison of their environmental requirements with the measured attributes of the environment at Gale Crater. We have already modeled a paleoenvironment with high habitability potential (HP) based upon chemistry, mineralogy and other geological evidence such as sedimentary structures and larger scale geomorphology [1]. In this report, we turn our attention to the present HP of the Yellowknife Bay area, including the importance of the physical environmental metrics such as atmospheric pressure, air and ground temperature, ionizing radiation, wind speed and direction, slope, etc.

Conrad, P. G.↗
NASA NTRS1999

LIfe: Past, Present and Future

Molecular methods of taxonomy and phylogeny have changed the way in which life on earth is viewed -- they have allowed us to transition from a eukaryote-centric view of the planet to one that is peculiarly prokarote-centric, containing three kingdoms, two of which are prokaryotic unicells.

biosignature↗
NASA NTRS2021

Anaerobic Expression and Purification of Holo-CCIS, an Artificial Iron-sulfur Protein

Iron-sulfur proteins are ubiquitous among all living organisms and are indispensable for almost all metabolic pathways ranging from photosynthesis, respiration, nitrogen, and carbon dioxide cycles. The iron-sulfur clusters primarily serve as electron acceptors and donors and transfer electrons to active sites of various enzymes, thus driving the energy metabolism. Prokaryotes like E. coli have ISC and SUF pathways that help in the assembly and maturation of iron-sulfur proteins. These ironsulfur proteins, especially with [4Fe-4S] clusters, are highly sensitive to molecular oxygen, and it would be advantageous if the de novo proteins and native proteins having iron-sulfur binding sites are expressed and isolated under anaerobic conditions. Bacterially assembled iron-sulfur proteins, when isolated and purified anaerobically, exhibit improved biochemical and biophysical stabilities in comparison to the counterparts expressed and purified aerobically and reconstituted under anaerobic conditions. This protocol outlines the expression and purification of the artificial protein, Coiled-Coil IronSulfur (CCIS). It may be deployed to both natural and artificial [4Fe-4S] proteins when heterologously expressed in E. coli.

Bhanu P. Jagilinki↗
NASA NTRS2003

Synchrony in human, mouse and bacterial cell cultures--a comparison

Growth characteristics of synchronous human MOLT-4, human U-937 and mouse L1210 cultures produced with a new minimally-disturbing technology were compared to each other and to synchronous Escherichia coli B/r. Based on measurements of cell concentrations during synchronous growth, synchrony persisted in similar fashion for all cells. Cell size and DNA distributions in the mammalian cultures also progressed synchronously and reproducibly for multiple cell cycles. The results demonstrate that unambiguous multi-cycle synchrony, critical for verifying the absence of significant growth imbalances induced by the synchronization procedure, is feasible with these cell lines, and possibly others.

Non-NASA Center↗
NASA NTRS1987

Symbiont acquisition as neoseme: origin of species and higher taxa

We examine the hypothesis that, in the origin of species and higher taxa of eukaryotes, symbiont acquisition followed by partner integration has been equivalent to neoseme appearance leading to speciation. The formation of stable symbiotic associations involves partner-surface recognition, behavioral and metabolic interaction, and, in some cases, gene product (RNA, protein) and genic (RNA, DNA) integration. This analysis is applied here to examples of neosemes that define specific taxa and to neosemes in plants, fungi, and animals that involve the appearance of new types of tissue. If this hypothesis is correct--if the origin of major genetic variation leading to speciation and even higher taxa may occur through symbiont acquisition and integration--then the analysis of "origins of species and higher taxa" becomes analogous to the study of microbial community ecology.

NASA Discipline Exobiology↗
NASA NTRS1994

The early evolution of life: solution to Darwin's dilemma

Recent studies of Precambrian fossils indicate that life on Earth originated earlier than assumed, microscopic life was prevalent in the Precambrian Eon, the tempo and mode of evolution during the Precambrian period were different from other periods, and that only the Precambrian fossil record can be used as evidence of early life. Implications for future research include directing the search for the origin of life away from the geological record, modification of hypotheses about molecular change, use of Precambrian microfossils in dating younger geological units, and progress in defining the nature of major events in early evolution.

Non-NASA Center↗
NASA NTRS1991

Kingdoms in turmoil

How should the world's living organisms be classified? Into how many kingdoms should they be grouped? Scientists have been grappling with these questions since the time of Aristotle, drawing on a broad base of biological characteristics for clues. The fossil record, visible traits of living organisms and, more recently, results from cell biology have all shaped theories of biological classification. But last year a new and controversial concept emerged: a classification of life based solely on molecular traits. The focal point of the controversy is a tree of life, or "phylogeny", devised by Carl Woese of the University of Illinois, Otto Kandler of the University of Munich and Mark Wheelis of the University of California. The tree is unusual because, unlike all previous schemes, it is constructed solely from biochemical data such as DNA sequences rather than a range of different organism characteristics. But that is not all. The scheme also challenges the idea that life on Earth is best divided into five kingdoms, with the main split being between bacteria and all other organisms. Woese and his colleagues create three main groupings by dividing the bacteria in two and unifying all other organisms.

NASA Program Exobiology↗
NASA NTRS1999

The influence of UV radiation on protistan evolution

Ultraviolet radiation has provided an evolutionary challenge to life on Earth. Recent increases in surficial ultraviolet B fluxes have focused attention on the role of UV radiation in protistan ecology, cancer, and DNA damage. Exploiting this new wealth of data, I examine the possibility that ultraviolet radiation may have played a significant role in the evolution of the first eukaryotes, that is, protists. Protists probably arose well before the formation of a significant ozone shield, and thus were probably subjected to substantial ultraviolet A, ultraviolet B, and ultraviolet C fluxes early in their evolution. Evolution consists of the generation of heritable variations and the subsequent selection of these variants. Ultraviolet radiation has played a role both as a mutagen and as a selective agent. In its role as a mutagen, it may have been crucial in the origin of sex and as a driver of molecular evolution. As a selective agent, its influence has been broad. Discussed in this paper are the influence of ultraviolet radiation on biogeography, photosynthesis, and desiccation resistance.

NASA Center ARC↗