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NASA NTRS1999

Membrane-Based Functions in the Origin of Cellular Life

Our objective is to help explain how the earliest ancestors of contemporary cells (protocells) performed their essential functions employing only the molecules available in the protobiological milieu. Our hypothesis is that vesicles, built of amphiphilic, membrane-forming materials, emerged early in protobiological evolution and served as precursors to protocells. We further assume that the cellular functions associated with contemporary membranes, such as capturing and, transducing of energy, signaling, or sequestering organic molecules and ions, evolved in these membrane environments. An alternative hypothesis is that these functions evolved in different environments and were incorporated into membrane-bound structures at some later stage of evolution. We focus on the application of the fundamental principles of physics and chemistry to determine how they apply to the formation of a primitive, functional cell. Rather than attempting to develop specific models for cellular functions and to identify the origin of the molecules which perform these functions, our goal is to define the structural and energetic conditions that any successful model must fulfill, therefore providing physico-chemical boundaries for these models. We do this by carrying out large-scale, molecular level computer simulations on systems of interest.

Chipot, Christophe↗

NASA NTRS2010

Solid-State, High Energy 2-Micron Laser Development for Space-Based Remote Sensing

Lidar (light detection and ranging) remote sensing enjoys the advantages of excellent vertical and horizontal resolution; pointing capability; a signal source independent from natural light; and control and knowledge of transmitted wavelength, pulse shape, and polarization and received polarization. Lidar in space is an emerging technology now being developing to fit applications where passive sensors cannot meet current measurement requirements. Technical requirements for space lidar are more demanding than for ground-based or airborne systems. Perhaps the most distinguishing characteristics of space lidars are the environmental requirements. Space lidar systems must be specially designed to survive the mechanical vibration loads of launch and operate in the vacuum of space where exposure to ionizing radiation limits the electronic components available. Finally, space lidars must be designed to be highly reliable because they must operate without the need for repair or adjustment. Lifetime requirements tend to be important drivers of the overall system design. The maturity of the required technologies is a key to the development of any space lidar system. NASA entered a new era in the 1990 s with the approval of several space-based remote sensing missions employing laser radar (lidar) techniques. Following the steps of passive remote sensing and then active radar remote sensing, lidar sensors were a logical next step, providing independence from natural light sources, and better spatial resolution and smaller sensor size than radar sensors. The shorter electromagnetic wavelengths of laser light also allowed signal reflectance from air molecules and aerosol particles. The smaller receiver apertures allowed the concept of scanning the sensor field of view. However, technical problems with several space-based lidar missions during that decade led to concern at NASA about the risk of lidar missions. An external panel was convened to make recommendations to NASA. Their report in 2000 strongly advocated that NASA maintain in-house laser and lidar capability, and that NASA should work to lower the technology risk for all future lidar missions. A multi-Center NASA team formulated an integrated NASA strategy to provide the technology and maturity of systems necessary to make Lidar/Laser systems viable for space-based study and monitoring of the Earth's atmosphere. In 2002 the NASA Earth Science Enterprise (ESE) and Office of Aerospace Technology (OAT) created the Laser Risk Reduction Program (LRRP) and directed NASA Langley Research Center (LaRC) and Goddard Space Flight Center to carry out synergistic and complementary research towards solid-state lasers/lidars developments for space-based remote sensing applications.

Singh, Upendra N.↗

NASA NTRS2015

Fabrication of Nanopipette Arrays for Biosensing

Method for providing a nanopipette array for biosensing applications. A thin substrate of anodizable metal ("AN-metal," such as Al, Mg, Zn, Ti, Ta and/or Nb) is anodized at temperature T=20-200.degree. C., chemical bath pH=4-6 and electrical potential 1-300 Volts, to produce an array of anodized nanopipette channels, having diameters 10-50 nm, with oxidized channel surfaces of thickness 5-20 nm. A portion of exposed non-oxidized AN-metal between adjacent nanopipette channels, of length 1-5 .mu.m, is etched away, exposing inner and outer surfaces of a nanopipette channel. A probe molecule, is deposited on one or both surfaces to provide biosensing capability for K(.gtoreq.1) target molecules. Target molecule presence, in an above-threshold concentration, in a fluid passed through or adjacent to a nanopipette channel, produces characteristic detection signals associated with the probe molecule site.

Meyyappan, Meyya↗

NASA NTRS1976

Millimeter emission lines in Orion A

During the course of a search of Orion A for signals from three large organic molecules, several millimeter-wave lines from known interstellar molecules were observed. Results are reported for observations of methanol (CH3OH), methyl cyanide (CH3CN), methyl acetylene (CH3CCH), acetaldehyde (CH3CHO) and (Si-29)O. Emission signals from two hydrogen recombination lines (H41-alpha and H42-alpha) detected from the H II region of Orion A are also reported. Negative results were obtained for several millimeter-wave transitions of ethylene oxide, acetone, and cyclopropenone.

Lovas, F. J.↗

NASA NTRS1992

Raman Lidar Measurements of Pinatubo Aerosols over Southeastern Kansas During November-december 1991

The eruptions of the Philippine volcano Pinatubo during June 1991 produced large amounts of stratospheric aerosols that could significantly affect earth's climate as well as trigger stratospheric ozone depletion through heterogeneous chemical reactions. Information regarding the physical and optical properties of these aerosols is required to quantify those effects. By measuring both the elastically backscattered signal and the inelastic signal produced by Raman scattering from nitrogen molecules, Raman lidar can provide some of this information. In this presentation we discuss Raman lidar measurements of the scattering ratio, backscattering, extinction, extinction/backscattering ratio, and optical thickness of the Pinatubo aerosols over southeastern Kansas made on 10 nights during November and December, 1991. The Raman lidar developed at GSFC is a trailer-based system which uses an XeF excimer laser to transmit light at 351 nm. The light backscattered by molecules and aerosols at this wavelength is detected as well as Raman scattered light from water vapor, nitrogen, and oxygen molecules. Since background skylights interfere with the detection of the Raman signals the data discussed in this paper were acquired only at night.

Ferrare, R. A.↗

NASA NTRS2004

Ultrasensitive Label-free Electronic Chip for DNA Analysis Using Carbon Nanotube Nanoelectrode Arrays

There is a strong need for faster, cheaper, and simpler methods for nucleic acid analysis in today s clinical tests. Nanotechnologies can potentially provide solutions to these requirements by integrating nanomaterials with biofunctionalities. Dramatic improvement in the sensitivity and multiplexing can be achieved through the high-degree miniaturization. Here, we present our study in the development of an ultrasensitive label-free electronic chip for DNA/RNA analysis based on carbon nanotube nanoelectrode arrays. A reliable nanoelectrode array based on vertically aligned multi-walled carbon nanotubes (MWNTs) embedded in a SiO2 matrix is fabricated using a bottom-up approach. Characteristic nanoelectrode behavior is observed with a low-density MWNT nanoelectrode array in measuring both the bulk and surface immobilized redox species. The open-end of MWNTs are found to present similar properties as graphite edge-plane electrodes, with a wide potential window, flexible chemical functionalities, and good biocompatibility. A BRCA1 related oligonucleotide probe with 18 bases is covalently functionalized at the open ends of the MWNTs and specifically hybridized with an oligonucleotide target as well as a PCR amplicon. The guanine bases in the target molecules are employed as the signal moieties for the electrochemical measurements. Ru(bpy)3(2+) mediator is used to further amplify the guanine oxidation signal. This technique has been employed for direct electrochemical detection of label-free PCR amplicon through specific hybridization with the BRCAl probe. The detection limit is estimated to be less than approximately 1000 DNA molecules, approaching the limit of the sensitivity by laser-based fluorescence techniques in DNA microarray. This system provides a general electronic platform for rapid molecular diagnostics in applications requiring ultrahigh sensitivity, high-degree of miniaturization, simple sample preparation, and low- cost operation.

Li, Jun↗

NASA NTRS2009

Enhancing SERS by Means of Supramolecular Charge Transfer

In a proposed method of sensing small quantities of molecules of interest, surface enhanced Raman scattering (SERS) spectroscopy would be further enhanced by means of intermolecular or supramolecular charge transfer. There is a very large potential market for sensors based on this method for rapid detection of chemical and biological hazards. In SERS, the Raman signals (vibrational spectra) of target molecules become enhanced by factors of the order of 108 when those molecules are in the vicinities of nanostructured substrate surfaces that have been engineered to have plasmon resonances that enhance local electric fields. SERS, as reported in several prior NASA Tech Briefs articles and elsewhere, has remained a research tool and has not yet been developed into a practical technique for sensing of target molecules: this is because the short range (5 to 20 nm) of the field enhancement necessitates engineering of receptor molecules to attract target molecules to the nanostructured substrate surfaces and to enable reliable identification of the target molecules in the presence of interferants. Intermolecular charge-transfer complexes have been used in fluorescence-, photoluminescence-, and electrochemistry-based techniques for sensing target molecules, but, until now, have not been considered for use in SERS-based sensing. The basic idea of the proposed method is to engineer receptor molecules that would be attached to nanostructured SERS substrates and that would interact with the target molecules to form receptor-target supramolecular charge-transfer complexes wherein the charge transfer could be photoexcited.

Wong, Eric↗

NASA NTRS1999

Genetic ablation of root cap cells in Arabidopsis

The root cap is increasingly appreciated as a complex and dynamic plant organ. Root caps sense and transmit environmental signals, synthesize and secrete small molecules and macromolecules, and in some species shed metabolically active cells. However, it is not known whether root caps are essential for normal shoot and root development. We report the identification of a root cap-specific promoter and describe its use to genetically ablate root caps by directing root cap-specific expression of a diphtheria toxin A-chain gene. Transgenic toxin-expressing plants are viable and have normal aerial parts but agravitropic roots, implying loss of root cap function. Several cell layers are missing from the transgenic root caps, and the remaining cells are abnormal. Although the radial organization of the roots is normal in toxin-expressing plants, the root tips have fewer cytoplasmically dense cells than do wild-type root tips, suggesting that root meristematic activity is lower in transgenic than in wild-type plants. The roots of transgenic plants have more lateral roots and these are, in turn, more highly branched than those of wild-type plants. Thus, root cap ablation alters root architecture both by inhibiting root meristematic activity and by stimulating lateral root initiation. These observations imply that the root caps contain essential components of the signaling system that determines root architecture.

Non-NASA Center↗

NASA NTRS2000

Evolutionary Cell Computing: From Protocells to Self-Organized Computing

On the path from inanimate to animate matter, a key step was the self-organization of molecules into protocells - the earliest ancestors of contemporary cells. Studies of the properties of protocells and the mechanisms by which they maintained themselves and reproduced are an important part of astrobiology. These studies also have the potential to greatly impact research in nanotechnology and computer science. Previous studies of protocells have focussed on self-replication. In these systems, Darwinian evolution occurs through a series of small alterations to functional molecules whose identities are stored. Protocells, however, may have been incapable of such storage. We hypothesize that under such conditions, the replication of functions and their interrelationships, rather than the precise identities of the functional molecules, is sufficient for survival and evolution. This process is called non-genomic evolution. Recent breakthroughs in experimental protein chemistry have opened the gates for experimental tests of non-genomic evolution. On the basis of these achievements, we have developed a stochastic model for examining the evolutionary potential of non-genomic systems. In this model, the formation and destruction (hydrolysis) of bonds joining amino acids in proteins occur through catalyzed, albeit possibly inefficient, pathways. Each protein can act as a substrate for polymerization or hydrolysis, or as a catalyst of these chemical reactions. When a protein is hydrolyzed to form two new proteins, or two proteins are joined into a single protein, the catalytic abilities of the product proteins are related to the catalytic abilities of the reactants. We will demonstrate that the catalytic capabilities of such a system can increase. Its evolutionary potential is dependent upon the competition between the formation of bond-forming and bond-cutting catalysts. The degree to which hydrolysis preferentially affects bonds in less efficient, and therefore less well-ordered, peptides is also critical to evolution of a non-genomic system. Based on these results, a new computational object called a "molnet" is defined. Like a neural network, it is formed of interconnected units that send "signals" to each other. Like molecules, neural networks have a specific function once their structure is defined. The difference between a molnet and traditional neural networks, is that input to molnets is not simply passed along and processed from input to output units, but rather it is utilized to form and break connections(bonds), and thus to form new structures. Molnets represent a powerful tool that can be used to understand the conditions under which chemical systems can form large molecules, such as proteins, and display ever more complex functions. This has direct applications, for example to the design of smart,synthetic fabrics. Additional information is contained in the original.

Colombano, Silvano↗

NASA NTRS1976

Chemistry between the stars

A unit is presented for the secondary school teacher of physics, chemistry, astronomy, or earth sciences. Included are a list of reference materials, teaching aids, and projects. Discussion questions and a glossary are also provided. Concepts developed are: the nature of interstellar space, spectroscopy, molecular signals from space and interstellar molecules and other areas of astronomy.

Gammon, R. H.↗

NASA NTRS1984

The far-infrared laser magnetic resonance spectrum of the SiH radical and determination of ground state parameters

The far-infrared laser magnetic resonance spectrum of the SiH radical in the v = O level of its X2Pi state has been recorded. The signals are rather weak. The molecules were generated in the reaction between fluorine atoms and SiH4. Rotational transitions have been detected in both 2Pi1/2 and 2Pi3/2 spin components but no fine structure transitions between the spin components were observed. Proton hyperfine splittings were resolved on some lines. The measurements have been analyzed, subjected to a least-squares fit using an effective Hamiltonian, and the appropriate molecular parameters determined. The weakness of the spectrum and the failure of attempts to power saturate favorable lines are both consistent with a small value for the electric dipole moment for SiH.

Brown, J. M.↗

NASA NTRS2000

Shape-dependent control of cell growth, differentiation, and apoptosis: switching between attractors in cell regulatory networks

Development of characteristic tissue patterns requires that individual cells be switched locally between different phenotypes or "fates;" while one cell may proliferate, its neighbors may differentiate or die. Recent studies have revealed that local switching between these different gene programs is controlled through interplay between soluble growth factors, insoluble extracellular matrix molecules, and mechanical forces which produce cell shape distortion. Although the precise molecular basis remains unknown, shape-dependent control of cell growth and function appears to be mediated by tension-dependent changes in the actin cytoskeleton. However, the question remains: how can a generalized physical stimulus, such as cell distortion, activate the same set of genes and signaling proteins that are triggered by molecules which bind to specific cell surface receptors. In this article, we use computer simulations based on dynamic Boolean networks to show that the different cell fates that a particular cell can exhibit may represent a preprogrammed set of common end programs or "attractors" which self-organize within the cell's regulatory networks. In this type of dynamic network model of information processing, generalized stimuli (e.g., mechanical forces) and specific molecular cues elicit signals which follow different trajectories, but eventually converge onto one of a small set of common end programs (growth, quiescence, differentiation, apoptosis, etc.). In other words, if cells use this type of information processing system, then control of cell function would involve selection of preexisting (latent) behavioral modes of the cell, rather than instruction by specific binding molecules. Importantly, the results of the computer simulation closely mimic experimental data obtained with living endothelial cells. The major implication of this finding is that current methods used for analysis of cell function that rely on characterization of linear signaling pathways or clusters of genes with common activity profiles may overlook the most critical features of cellular information processing which normally determine how signal specificity is established and maintained in living cells. Copyright 2000 Academic Press.

Review↗

NASA NTRS1989

Continuous electron stimulated desorption using a ZrO2/Ag permeation membrane

During the development of an atomic oxygen beam generator for laboratory simulation of the atmospheric conditions in low earth orbit, a new technique for performing electron stimulated desorption (ESD) in a continuous manner has been developed. In this technique, oxygen permeates through an Ag membrane at elevated temperature thereby providing a continuous supply of oxygen atoms to the 1000-A ZrO2 coating at the vacuum interface. ESD then results in a large peak of neutral O2 molecules which ultimately decay into steady-state desorption. The ESD signal is linear with respect to primary beam flux (0.035 O2 molecules per electron at a primary beam energy of 1 keV) but nonlinear with respect to primary beam energy.

Outlaw, R. A.↗

NASA NTRS2001

Visualization of Microbial Biomarkers by Scanning Electron Microscopy

We are developing tools to link the biochemical structure of selected biomarkers with putative biogenic structures observed in mineralized samples. The detection of evidence of life on Mars and other planets will rely on methods that can discriminate compounds formed exclusively by living organisms. While biogenic compounds, such as amino acids and nucleotides have been discovered in extraterrestrial sources, such as meteorites and comets, their formation can be explained by abiotic means. The formation of cellular structures, or more elaborate organic molecules, such as complex lipids, proteins or nucleic acids, however, is strongly correlated to the presence of even the most primitive life processes. Recent evidence lends support to the hypothesis that life may have once existed on Mars. Carbonate globules and ppm concentrations of polycyclic aromatic hydrocarbons (PAHs) have been described in ALH84001, a meteorite originating from Mars ejecta captured by Earth over 13,000 years ago. The localized high concentration of PAHs that follow an increasing gradient from the intact fusion crust towards the interior corresponds to microgram quantities of hydrocarbon. Even though ALH84001 and other similar meteorites have withstood the forces capable of ejecting rock through Mars' escape velocity, upon entering Earth's atmosphere, their core temperatures are likely not to have been raised significantly, as evidenced by the survival of remanent magnetic signatures. Ideal biomarkers of ancient or modern biological life would include molecules that are (or were) pervasive and highly resistant to degradation. Also, requisite methods of detection should be simple, extremely sensitive and broadly inclusive (NASA SP-530). Lipopolysaccharide (LPS), peptidoglycan or pseudopeptidoglycan and beta-glucan are microbial cell wall components which together cover the entire microbial spectrum of eubacteria, archea and fungi. They are all remarkably resistant to thermal degradation. Fortunately, many antimicrobial defense systems of higher organisms require sensitive detection to combat microbial pathogens. We employ here the primitive immune system of the evolutionarily ancient horseshoe crab, Limulus polyphemus. This species relies on multi-enzyme signal amplification detection of cell wall molecules and they can be applied to the development of useful detectors of life. An extension of this work includes the visualization of microbial signatures by labeling LAL components with chromogenic or electron dense markers. The protein Limulus Anti-LPS Factor (LALF) has an extremely high affinity for LPS. By coupling LALF binding with colloidal gold labels we demonstrate a correlation of the structures visible by electron microscopy with biochemical evidence of microbial cell wall materials. Pure silica particles were mixed with cultures of E. coli (10(exp 6) cfu/mL). Samples were washed sequentially with buffered saline, LALF, antibody to LALF and finally colloidal gold-labeled Protein A. Negative controls were not exposed to E. coli but received identical treatment otherwise. Samples were coated with carbon and imaged on a JEOL JSM-840 scanning electron microscope with LaB6 source in the back scatter mode with the JEOL annular back scatter detector. 20 nm-scale black spots in this contrast-reversed image originate from electrons back-scattered by gold atoms. Negative controls did not give any signal. Future work will expand application of this technique to soil simulants and mineralized rock samples.

Wainwright, Norman R.↗

NASA NTRS2004

A New Way to Measure Cirrus Ice Water Content by Using Ice Raman Scatter with Raman Lidar

High and cold cirrus clouds mainly contain irregular ice crystals, such as, columns, hexagonal plates, bullet rosettes, and dendrites, and have different impacts on the climate system than low-level clouds, such as stratus, stratocumulus, and cumulus. The radiative effects of cirrus clouds on the current and future climate depend strongly on cirrus cloud microphysical properties including ice water content (IWC) and ice crystal sizes, which are mostly an unknown aspect of cinus clouds. Because of the natural complexity of cirrus clouds and their high locations, it is a challenging task to get them accurately by both remote sensing and in situ sampling. This study presents a new method to remotely sense cirrus microphysical properties by using ice Raman scatter with a Raman lidar. The intensity of Raman scattering is fundamentally proportional to the number of molecules involved. Therefore, ice Raman scattering signal provides a more direct way to measure IWC than other remote sensing methods. Case studies show that this method has the potential to provide essential information of cirrus microphysical properties to study cloud physical processes in cirrus clouds.

Wang, Zhien↗

NASA NTRS2009

Selective functionalization of carbon nanotube tips allowing fabrication of new classes of nanoscale sensing and manipulation tools

Embodiments in accordance with the present invention relate to techniques for the growth and attachment of single wall carbon nanotubes (SWNT), facilitating their use as robust and well-characterized tools for AFM imaging and other applications. In accordance with one embodiment, SWNTs attached to an AFM tip can function as a structural scaffold for nanoscale device fabrication on a scanning probe. Such a probe can trigger, with nanometer precision, specific biochemical reactions or conformational changes in biological systems. The consequences of such triggering can be observed in real time by single-molecule fluorescence, electrical, and/or AFM sensing. Specific embodiments in accordance with the present invention utilize sensing and manipulation of individual molecules with carbon nanotubes, coupled with single-molecule fluorescence imaging, to allow observation of spectroscopic signals in response to mechanically induced molecular changes. Biological macromolecules such as proteins or DNA can be attached to nanotubes to create highly specific single-molecule probes for investigations of intermolecular dynamics, for assembling hybrid biological and nanoscale materials, or for developing molecular electronics. In one example, electrical wiring of single redox enzymes to carbon nanotube scanning probes allows observation and electrochemical control over single enzymatic reactions by monitoring fluorescence from a redox-active cofactor or the formation of fluorescent products. Enzymes ''nanowired'' to the tips of carbon nanotubes in accordance with embodiments of the present invention, may enable extremely sensitive probing of biological stimulus-response with high spatial resolution, including product-induced signal transduction.

Wade, Lawrence A.↗

NASA NTRS2012

Receivers Gather Data for Climate, Weather Prediction

Signals from global positioning system (GPS) satellites are now being used for more than just location and navigation information. By looking at the radio waves from GPS satellites, a technology developed at NASA s Jet Propulsion Laboratory (JPL) not only precisely calculates its position, but can also use a technique known as radio occultation to help scientists study the Earth s atmosphere and gravity field to improve weather forecasts, monitor climate change, and enhance space weather research. The University Corporation for Atmospheric Research (UCAR), a nonprofit group of universities in Boulder, Colorado, compares radio occultation to the appearance of a pencil when viewed though a glass of water. The water molecules change the path of visible light waves so that the pencil appears bent, just like molecules in the air bend GPS radio signals as they pass through (or are occulted by) the atmosphere. Through measurements of the amount of bending in the signals, scientists can construct detailed images of the ionosphere (the energetic upper part of the atmosphere) and also gather information about atmospheric density, pressure, temperature, and moisture. Once collected, this data can be input into weather forecasting and climate models for weather prediction and climate studies. Traditionally, such information is obtained through the use of weather balloons. In 1998, JPL started developing a new class of GPS space science receivers, called Black Jack, that could take precise measurements of how GPS signals are distorted or delayed along their way to the receiver. By 2006, the first demonstration of a GPS radio occultation constellation was launched through a collaboration among Taiwan s National Science Council and National Space Organization, the U.S. National Science Foundation, NASA, the National Oceanic and Atmospheric Administration (NOAA), and other Federal entities. Called the Constellation Observing System for Meteorology, Ionosphere, and Climate (COSMIC), JPL was responsible for designing COSMIC s primary instrument - based on its revolutionary Black Jack receiver.

Source record↗

NASA NTRS2004

Mechanotransduction through Integrins

The goal of this project was to characterize the molecular mechanism by which cells recognize and respond to physical forces in their local environment. The project was based on the working hypothesis that cells sense mechanical stresses through cell surface integrin receptors and through their interconnections with the underlying cytoskeleton. Work completed and published in past funding period had provided direct support for this hypothesis. In particular, we demonstrated that application of mechanical stresses to activated integrin receptors (but not inactive integrins or other control transmembrane receptors) resulted in stress-dependent activation of the CAMP signaling pathway leading to gene transcription. We also showed that this form of mechanotransduction requires activation of heterotrimeric G proteins. In this grant, our specific aims included: 1) to characterize the signal processing capabilities of different integrins and other cell surface receptors, 2) to identify heterotrimeric G proteins that mediate CAMP signaling by stresses applied to integrins, 3) to identify molecules that mediate transmembrane mechanochemical coupling between integrins and G proteins, and 4) to use genome-wide gene expression profiling techniques to identify other genes and signaling pathways that are activated by mechanical forces transmitted over specific cell surface receptors. Elucidation of the mechanism by which cells sense mechanical stresses through integrins and translate them into a biochemical response should help us to understand the molecular basis of the cellular response to gravity as well as many other forms of mechanosensation and tissue regulation.

Ingber, Donald↗