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At least 109 records · Page 6

Amplifying Electrochemical Indicators

Dendrimeric reporter compounds have been invented for use in sensing and amplifying electrochemical signals from molecular recognition events that involve many chemical and biological entities. These reporter compounds can be formulated to target specific molecules or molecular recognition events. They can also be formulated to be, variously, hydrophilic or amphiphilic so that they are suitable for use at interfaces between (1) aqueous solutions and (2) electrodes connected to external signal-processing electronic circuits. The invention of these reporter compounds is expected to enable the development of highly miniaturized, low-power-consumption, relatively inexpensive, mass-producible sensor units for diverse applications.

Fan, Wenhong↗

Laser optogalvanic spectroscopy of molecules

In laser optogalvanic (LOG) spectroscopy, a tunable laser is used to probe the spectral characteristics of atomic or molecular species within an electrical discharge in a low pressure gas. Optogalvanic signals arise when the impedance of the discharge changes in response to the absorption of laser radiation. The technique may, therefore, be referred to as impedance spectroscopy. This change in impedance may be monitored as a change in the voltage across the discharge tube. LOG spectra are recorded by scanning the wavelength of a chopped CW dye laser while monitoring the discharge voltage with a lock-in amplifier. LOG signals are obtained if the laser wavelength matches a transition in a species present in the discharge (or flame), and if the absorption of energy in the laser beam alters the impedance of the discharge. Infrared LOG spectroscopy of molecules has been demonstrated and may prove to be the most productive application in the field of optogalvanic techniques.

Webster, C. R.↗

Skeletal phenotype of growing transgenic mice that express a function-perturbing form of beta1 integrin in osteoblasts

Skeletal modeling entails the deposition of large amounts of extracellular matrix (ECM) to form structures tailored to withstand increasing mechanical loads during rapid growth. Specific ECM molecules bind to integrin receptors on the cell surface, thereby triggering a cascade of signaling events that affect critical cell functions. To evaluate the role of integrins during skeletal growth, transgenic mice were engineered to express a function-perturbing fragment of beta1 integrin consisting of the transmembrane domain and cytoplasmic tail under the control of the osteocalcin promoter (TG mice). Thus, transgene expression was targeted to mature cells of the osteoblast lineage, and herein we show that cultured cells resembling osteocytes from 90-day-old TG mice display impaired adhesion to collagen I, a ligand for beta1 integrin. To determine the influence of beta1 integrin on bones that are responsible for providing structural support during periods of rapid growth, we examined the phenotype of the appendicular skeleton in TG mice compared to wild type (WT) mice. According to radiographs, bones from mice of both genotypes between 14 and 90 days of age appeared similar in gross structure and density, although proximal tibiae from 35-90 days old TG mice were less curved than those of WT mice (72-92% TG/WT). Although there were only mild and transient differences in absolute bone mass and strength, once normalized to body mass, the tibial dry mass (79.1% TG/WT females), ash mass (78.5% TG/WT females), and femoral strength in torsion (71.6% TG/WT females) were reduced in TG mice compared to WT mice at 90 days of age. Similar effects of genotype on bone mass and curvature were observed in 1-year-old retired breeders, indicating that these phenotypic differences between TG and WT mice were stable well into adulthood. Effects of genotype on histomorphometric indices of cancellous bone turnover were minimal and evident only transiently during growth, but when present they demonstrated differences in osteoblast rather than osteoclast parameters. Together, these results suggest that integrin signals generated during growth enhance the acquisition of a skeletal mass, structure, and strength to withstand the mechanical loads generated by weight-bearing.

Antigens, CD29/genetics/metabolism↗

VLA searches for formaldehyde and cyanoacetylene emission from comet P/Brorsen-Metcalf (1989o)

In Sep. 1989, Very Large Array (VLA) searches were conducted for both formaldehyde (H2CO) and cyanoacetylene (HC3N) emission from Comet P/Brorsen-Metcalf (1989o). In early July, this comet was outgassing strongly at a brighter than average preperihelion rate, and the hope was that it would continue to do so through perihelion. By observing the 1 sub 11 to 1 sub 10 transition of H2CO at 4,829.659 MHz (6 cm lambda) during the phase of maximum outgassing, it was expected that spectra would be collected with a high signal to noise ratio which would show the temporal behavior of formaldehyde relative to other molecules. During this same observation period, a search was also made for the J-1-0, F-2-1 rotational transition of cyanoacetylene at 9098.3321 MHz (3.3 cm lambda).

Snyder, Lewis E.↗

Oxidative stress inhibits caveolin-1 palmitoylation and trafficking in endothelial cells

During normal and pathological conditions, endothelial cells (ECs) are subjected to locally generated reactive oxygen species, produced by themselves or by other vessel wall cells. In excess these molecules cause oxidative injury to the cell but at moderate levels they might modulate intracellular signalling pathways. We have investigated the effect of oxidative stress on the palmitoylation and trafficking of caveolin-1 in bovine aortic ECs. Exogenous H2O2 did not alter the intracellular localization of caveolin-1 in ECs. However, metabolic labelling experiments showed that H2O2 inhibited the trafficking of newly synthesized caveolin-1 to membrane raft domains. Several mechanisms potentially responsible for this inhibition were examined. Impairment of caveolin-1 synthesis by H2O2 was not responsible for diminished trafficking. Similarly, the inhibition was independent of H2O2-induced caveolin-1 phosphorylation as shown by the markedly different concentration dependences. We tested the effect of H2O2 on palmitoylation of caveolin-1 by the incorporation of [3H]palmitic acid. Exposure of ECs to H2O2 markedly inhibited the palmitoylation of caveolin-1. Comparable inhibition was observed after treatment of cells with H2O2 delivered either as a bolus or by continuous delivery with glucose and glucose oxidase. Kinetic studies showed that H2O2 did not alter the rate of caveolin-1 depalmitoylation but instead decreased the 'on-rate' of palmitoylation. Together these results show for the first time the modulation of protein palmitoylation by oxidative stress, and suggest a cellular mechanism by which stress might influence caveolin-1-dependent cell activities such as the concentration of signalling proteins and cholesterol trafficking.

NASA Discipline Cell Biology↗

Modeled microgravity inhibits apoptosis in peripheral blood lymphocytes

Microgravity interferes with numerous lymphocyte functions (expression of cell surface molecules, locomotion, polyclonal and antigen-specific activation, and the protein kinase C activity in signal transduction). The latter suggests that gravity may also affect programmed cell death (PCD) in lymphocyte populations. To test this hypothesis, we investigated spontaneous, activation- and radiation-induced PCD in peripheral blood mononuclear cells exposed to modeled microgravity (MMG) using a rotating cell culture system. The results showed significant inhibition of radiation- and activation-induced apoptosis in MMG and provide insights into the potential mechanisms of this phenomenon.

NASA Discipline Cell Biology↗

Modeled Microgravity Inhibits Apoptosis in Peripheral Blood Lymphocytes

Microgravity interferes with numerous lymphocyte functions (expression of cell surface molecules, locomotion, polyclonal and antigen-specific activation, and the protein kinase C activity in signal transduction). The latter suggests that gravity may also affect programmed cell death (PCD) in lymphocyte populations. To test this hypothesis, we investigated spontaneous, activation- and radiation-induced PCD in peripheral blood mononuclear cells (PBMC) exposed to modeled microgravity using a rotating cell culture system. The results showed significant inhibition of radiation- and activation-induced apoptosis in modeled microgravity and provide insights into the potential mechanisms of this phenomenon.

Risin, Diana↗

Molecular Composition Analysis of Distant Targets

This document is the Final Report for NASA Innovative Advanced Concepts (NIAC) Phase I Grant 15-NIAC16A-0145, titled Molecular Composition Analysis of Distant Targets. The research was focused on developing a system concept for probing the molecular composition of cold solar system targets, such as Asteroids, Comets, Planets and Moons from a distant vantage, for example from a spacecraft that is orbiting the target (Hughes et al., 2015). The orbiting spacecraft is equipped with a high-power laser, which is run by electricity from photovoltaic panels. The laser is directed at a spot on the target. Materials on the surface of the target are heated by the laser beam, and begin to melt and then evaporate, forming a plume of asteroid molecules in front of the heated spot. The heated spot glows, producing blackbody illumination that is visible from the spacecraft, via a path through the evaporated plume. As the blackbody radiation from the heated spot passes through the plume of evaporated material, molecules in the plume absorb radiation in a manner that is specific to the rotational and vibrational characteristics of the specific molecules. A spectrometer aboard the spacecraft is used to observe absorption lines in the blackbody signal. The pattern of absorption can be used to estimate the molecular composition of materials in the plume, which originated on the target. Focusing on a single spot produces a borehole, and shallow subsurface profiling of the targets bulk composition is possible. At the beginning of the Phase I research, the estimated Technology Readiness Level (TRL) of the system was TRL-1. During the Phase I research, an end-to-end theoretical model of the sensor system was developed from first principles. The model includes laser energy and optical propagation, target heating, melting and evaporation of target material, plume density, thermal radiation from the heated spot, molecular cross section of likely asteroid materials, and estimation of the absorption profile at a distant spectrometer. Results obtained by executing simulations based on the model provide compelling evidence that the concept of remote laser evaporative molecular absorption spectroscopy is feasible. In this document, technical details of the model are presented, and results of simulations are described that indicate the utility of the proposed sensor system. Additionally, an asteroid rendezvous mission is analyzed, with a survey of system requirements to accomplish molecular composition analysis of the asteroid. Based on positive theoretical results obtained during Phase I, the estimated TRL of the system is now TRL-2. This document also describes potential future research and experimentation that could push the system to TRL-4 within 2 years. Steps required for construction of a laboratory prototype are described. An experiment to test predictions of the theory is described, based on the laboratory prototype setup.

asteroid↗

Modular and Stochastic Approaches to Molecular Pathway Models of ATM, TGF beta, and WNT Signaling

Deterministic pathway models that describe the biochemical interactions of a group of related proteins, their complexes, activation through kinase, etc. are often the basis for many systems biology models. Low dose radiation effects present a unique set of challenges to these models including the importance of stochastic effects due to the nature of radiation tracks and small number of molecules activated, and the search for infrequent events that contribute to cancer risks. We have been studying models of the ATM, TGF -Smad and WNT signaling pathways with the goal of applying pathway models to the investigation of low dose radiation cancer risks. Modeling challenges include introduction of stochastic models of radiation tracks, their relationships to more than one substrate species that perturb pathways, and the identification of a representative set of enzymes that act on the dominant substrates. Because several pathways are activated concurrently by radiation the development of modular pathway approach is of interest.

Cucinotta, Francis A.↗

Rac-1 and Raf-1 kinases, components of distinct signaling pathways, activate myotonic dystrophy protein kinase

Myotonic dystrophy protein kinase (DMPK) is a serine-threonine protein kinase encoded by the myotonic dystrophy (DM) locus on human chromosome 19q13.3. It is a close relative of other kinases that interact with members of the Rho family of small GTPases. We show here that the actin cytoskeleton-linked GTPase Rac-1 binds to DMPK, and coexpression of Rac-1 and DMPK activates its transphosphorylation activity in a GTP-sensitive manner. DMPK can also bind Raf-1 kinase, the Ras-activated molecule of the MAP kinase pathway. Purified Raf-1 kinase phosphorylates and activates DMPK. The interaction of DMPK with these distinct signals suggests that it may play a role as a nexus for cross-talk between their respective pathways and may partially explain the remarkable pleiotropy of DM.

NASA Discipline Musculoskeletal↗

Modulation of lens cell adhesion molecules by particle beams

Cell adhesion molecules (CAMs) are proteins which anchor cells to each other and to the extracellular matrix (ECM), but whose functions also include signal transduction, differentiation, and apoptosis. We are testing a hypothesis that particle radiations modulate CAM expression and this contributes to radiation-induced lens opacification. We observed dose-dependent changes in the expression of beta 1-integrin and ICAM-1 in exponentially-growing and confluent cells of a differentiating human lens epithelial cell model after exposure to particle beams. Human lens epithelial (HLE) cells, less than 10 passages after their initial culture from fetal tissue, were grown on bovine corneal endothelial cell-derived ECM in medium containing 15% fetal bovine serum and supplemented with 5 ng/ml basic fibroblast growth factor (FGF-2). Multiple cell populations at three different stages of differentiation were prepared for experiment: cells in exponential growth, and cells at 5 and 10 days post-confluence. The differentiation status of cells was characterized morphologically by digital image analysis, and biochemically by Western blotting using lens epithelial and fiber cell-specific markers. Cultures were irradiated with single doses (4, 8 or 12 Gy) of 55 MeV protons and, along with unirradiated control samples, were fixed using -20 degrees C methanol at 6 hours after exposure. Replicate experiments and similar experiments with helium ions are in progress. The intracellular localization of beta 1-integrin and ICAM-1 was detected by immunofluorescence using monoclonal antibodies specific for each CAM. Cells known to express each CAM were also processed as positive controls. Both exponentially-growing and confluent, differentiating cells demonstrated a dramatic proton-dose-dependent modulation (upregulation for exponential cells, downregulation for confluent cells) and a change in the intracellular distribution of the beta 1-integrin, compared to unirradiated controls. In contrast, there was a dose-dependent increase in ICAM-1 immunofluorescence in confluent, but not exponentially-growing cells. These results suggest that proton irradiation downregulates beta 1-integrin and upregulates ICAM-1, potentially contributing to cell death or to aberrant differentiation via modulation of anchorage and/or signal transduction functions. Quantification of the expression levels of the CAMs by Western analysis is in progress.

NASA Discipline Radiation Health↗

Xwnt-8 and lithium can act upon either dorsal mesodermal or neurectodermal cells to cause a loss of forebrain in Xenopus embryos

When Xenopus gastrulae are made to misexpress Xwnt-8, or are exposed to lithium ions, they develop with a loss of anterior structures. In the current study, we have characterized the neural defects produced by either Xwnt-8 or lithium and have examined potential cellular mechanisms underlying this anterior truncation. We find that the primary defect in embryos exposed to lithium at successively earlier stages during gastrulation is a progressive rostral to caudal deletion of the forebrain, while hindbrain and spinal regions of the CNS remain intact. Misexpression of Xwnt-8 during gastrulation produces an identical loss of forebrain. Our results demonstrate that lithium and Wnts can act upon either prospective neural ectodermal cells, or upon dorsal mesodermal cells, to cause a loss of anterior pattern. Specifically, ectodermal cells isolated from lithium- or Wnt-exposed embryos are unable to form anterior neural tissue in response to inductive signals from normal dorsal mesoderm. In addition, although dorsal mesodermal cells from lithium- or Wnt-exposed embryos are specified properly, and produce normal levels of the anterior neural inducing molecules noggin and chordin, they show a greatly reduced capacity to induce anterior neural tissue in conjugated ectoderm. Taken together, our results are consistent with a model in which Wnt- or lithium-mediated signals can induce either mesodermal or ectodermal cells to produce a dominant posteriorizing morphogen which respecifies anterior neural tissue as posterior.

Non-NASA Center↗

Demonstration of Frequency-Scanning Burst-mode Filtered Rayleigh Scattering for Multi-Parameter Gas-Phase Measurements

Filtered Rayleigh scattering (FRS) is a diagnostic technique used for measurements of macroscopic properties of a gas. The FRS signal is passed through a narrow bandwidth molecular filter, commonly gaseous iodine, to measure the broadening and Doppler-shift of the scattering light from the molecules of the gas flow. However, in single-frequency, intensity-based approaches, the effects of pressure, temperature, and velocity can be difficult to deconvolve. To overcome this challenge, frequency-scanning FRS has been used to quasi-spectrally resolve the signal. In this work, a frequency-scanned, burst-mode laser was used to perform scanning FRS in an underexpanded jet to measure temperature, pressure, and radial velocity at measurement rates up to 1 kHz which is ~100,000 times faster than prior implementations of scanning FRS.

filtered Rayleigh scattering↗

Measurements of the Vertical Structure of Aerosols and Clouds Over the Ocean Using Micro-Pulse LIDAR Systems

The determination of the vertical distribution of aerosols and clouds over the ocean is needed for accurate retrievals of ocean color from satellites observations. The presence of absorbing aerosol layers, especially at altitudes above the boundary layer, has been shown to influence the calculation of ocean color. Also, satellite data must be correctly screened for the presence of clouds, particularly cirrus, in order to measure ocean color. One instrument capable of providing this information is a lidar, which uses pulses of laser light to profile the vertical distribution of aerosol and cloud layers in the atmosphere. However, lidar systems prior to the 1990s were large, expensive, and not eye-safe which made them unsuitable for cruise deployments. During the 1990s the first small, autonomous, and eye-safe lidar system became available: the micro-pulse lidar, or MPL. The MPL is a compact and eye-safe lidar system capable of determining the range of aerosols and clouds by firing a short pulse of laser light (523 nm) and measuring the time-of-flight from pulse transmission to reception of a returned signal. The returned signal is a function of time, converted into range using the speed of light, and is proportional to the amount of light backscattered by atmospheric molecules (Rayleigh scattering), aerosols, and clouds. The MPL achieves ANSI eye-safe standards by sending laser pulses at low energy (micro-J) and expanding the beam to 20.32 cm in diameter. A fast pulse-repetition-frequency (2500 Hz) is used to achieve a good signal-to-noise, despite the low output energy. The MPL has a small field-of-view (< 100 micro-rad) and signals received with the instrument do not contain multiple scattering effects. The MPL has been used successfully at a number of long-term sites and also in several field experiments around the world.

Welton, Ellsworth J.↗

Intensity and pressure dependence of resonance fluorescence of OH induced by a tunable UV laser

The intensity and pressure dependence of the fluorescence spectrum of OH in the presence of N2 and H2O molecules was studied. Saturation of the absorption transition was observed at low pressures, and the corresponding fluorescence signal was found to vary as the square root of the exciting intensity. This observed dependence agreed with the predicted dependence which took into account the presence of laser modes in the spectrum of the exciting radiation. With full laser power incident, a saturation parameter as high as 3 x 10 to the 5th was observed. The fluorescence spectrum was found to peak at 3145 and at 3090 A, with the relative peak intensities dependent upon gas pressures and upon the particular rotational electronic transition used for excitation. It is concluded that vibrational relaxation of the electronically excited OH due to water vapor in the system plays a dominant role in determining the observed fluorescence spectrum.

Killinger, D. K.↗

Measurement of temperature and density fluctuations in turbulence using an ultraviolet laser

Noninvasive measurement of density and temperature fluctuations in turbulent air flow was examined. The approach used fluorescence of oxygen molecules which are selectively excited by a tunable vacuum ultraviolet laser beam. The strength of the fluorescence signal and its dependence on laser wavelength vary with the density and temperature of the air in the laser beam. Because fluorescence can be detected at 90 degrees from the beam propagation direction, spatial resolution in three dimensions, rather than path-integrated measurements can be achieved. With spatial resolutions of the order of a millimeter and at supersonic air velocities it is necessary to perform each measurement in a time of the order of a microsecond; this is possible by by using laser pulses of ten nanosecond duration. In this method atmospheric O2 is excited by the emission of a tunable ArF excimer laser, and the fluorescence, which spans the 210 to 420 range, is detected by an ultraviolet phototube.

Massey, G. A.↗

Free-falling Crystals: Biological Macromolecular Crystal Growth Studies in Low Earth Orbit

Spacecraft orbiting the earth experience a reduced acceleration environment due to being in a state of continuous free-fall. This state colloquially termed microgravity, has produced improved X-ray diffraction quality crystals of biological macromolecules. Improvements in X-ray diffraction resolution (detail) or signal to noise, provide greater detail in the three-dimensional molecular structure providing information about the molecule, how it works, how to improve its function or how to impede it. Greater molecular detail obtained by crystallization in microgravity, has important implications for structural biology. In this article we examine the theories behind macromolecule crystal quality improvement in microgravity using results obtained from studies with the model protein, chicken egg white lysozyme.

Judge, Russell A.↗

All-Solid-State 2.45-to-2.78-THz Source

Sources in the THz range are required in order for NASA to implement heterodyne instruments in this frequency range. The source that has been demonstrated here will be used for an instrument on the SOFIA platform as well as for upcoming astrophysics missions. There are currently no electronic sources in the 2 3- THz frequency range. An electronically tunable compact source in this frequency range is needed for lab spectroscopy as well as for compact space-deployable heterodyne receivers. This solution for obtaining useful power levels in the 2 3- THz range is based on utilizing power-combined multiplier stages. Utilizing power combining, the input power can be distributed between different multiplier chips and then recombined after the frequency multiplication. A continuous wave (CW) coherent source covering 2.48 2.75 THz, with greater than 10 percent instantaneous and tuning bandwidth, and having l 14 W of output power at room temperature, has been demonstrated. This source is based on a 91.8 101.8-GHz synthesizer followed by a power amplifier and three cascaded frequency triplers. It demonstrates that purely electronic solid-state sources can generate a useful amount of power in a region of the electromagnetic spectrum where lasers (solid-state or gas) were previously the only available coherent sources. The bandwidth, agility, and operability of this THz source has enabled wideband, high-resolution spectroscopic measurements of water, methanol, and carbon monoxide with a resolution and signal-to-noise ratio unmatched by other existing systems, providing new insight in the physics of these molecules. Further - more, the power and optical beam quality are high enough to observe the Lamb-dip effect in water. The source frequency has an absolute accuracy better than 1 part in 1012, and the spectrometer achieves sub-Doppler frequency resolution better than 1 part in 108. The harmonic purity is better than 25 dB. This source can serve as a local oscillator for a variety of heterodyne systems, and can be used as a method for precision control of more powerful but much less frequency-agile quantum mechanical terahertz sources.

Mehdi, Imran↗