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At least 109 records · Page 6

System and method for monitoring cellular activity

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

Bearman, Gregory H.↗

System and method for monitoring cellular activity

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

Bearman, Gregory H.↗

Cellular reflectarray antenna and method of making same

A method of manufacturing a cellular reflectarray antenna arranged in an m by n matrix of radiating elements for communication with a satellite includes steps of determining a delay .phi.m,n for each of said m by n matrix of elements of said cellular reflectarray antenna using sub-steps of: determining the longitude and latitude of operation, determining elevation and azimuth angles of the reflectarray with respect to the satellite and converting theta.sub.0 (.theta..sub.0) and phi.sub.0 (.phi..sub.0), determining .DELTA..beta..sub.m,n, the pointing vector correction, for a given inter-element spacing and wavelength, determining .DELTA..phi..sub.m,n, the spherical wave front correction factor, for a given radius from the central element and/or from measured data from the feed horn; and, determining a delay .phi.m,n for each of said m by n matrix of elements as a function of .DELTA..beta..sub.m,n and .DELTA..phi..sub.m,n..

Romanofsky, Robert R↗

Cellular reflectarray antenna and method of making same

A method of manufacturing a cellular reflectarray antenna arranged in an m by n matrix of radiating elements for communication with a satellite includes steps of determining a delay .phi.m,n for each of said m by n matrix of elements of said cellular reflectarray antenna using sub-steps of: determining the longitude and latitude of operation, determining elevation and azimuth angles of the reflectarray with respect to the satellite and converting theta.sub.0 (.theta..sub.0) and phi.sub.0 (.phi..sub.0), determining .DELTA..beta..sub.m,n, the pointing vector correction, for a given inter-element spacing and wavelength, determining .DELTA..phi..sub.m,n, the spherical wave front correction factor, for a given radius from the central element and/or from measured data from the feed horn; and, determining a delay .phi.m,n for each of said m by n matrix of elements as a function of .DELTA..beta..sub.m,n and .DELTA..phi..sub.m,n.

Romanofsky, Robert R↗

Ground Testing of the EMCS Seed Cassette for Biocompatibility with the Cellular Slime Mold, Dictyostelium Discoideum

The European Modular Cultivation System, EMCS, was developed by ESA for plant experiments. To expand the use of flight verified hardware for various model organisms, we performed ground experiments to determine whether ARC EMCS Seed Cassettes could be adapted for use with cellular slime mold for future space flight experiments. Dictyostelium is a cellular slime mold that can exist both as a single-celled independent organism and as a part of a multicellular colony which functions as a unit (pseudoplasmodium). Under certain stress conditions, individual amoebae will aggregate to form multicellular structures. Developmental pathways are very similar to those found in Eukaryotic organisms, making this a uniquely interesting organism for use in genetic studies. Dictyostelium has been used as a genetic model organism for prior space flight experiments. Due to the formation of spores that are resistant to unfavorable conditions such as desiccation, Dictyostelium is also a good candidate for use in the EMCS Seed Cassettes. The growth substratum in the cassettes is a gridded polyether sulfone (PES) membrane. A blotter beneath the PES membranes contains dried growth medium. The goals of this study were to (1) verify that Dictyostelium are capable of normal growth and development on PES membranes, (2) develop a method for dehydration of Dictyostelium spores with successful recovery and development after rehydration, and (3) successful mock rehydration experiments in cassettes. Our results show normal developmental progression in two strains of Dictyostelium discoideum on PES membranes with a bacterial food source. We have successfully performed a mock rehydration of spores with developmental progression from aggregation to slug formation, and production of morphologically normal spores within 9 days of rehydration. Our results indicate that experiments on the ISS using the slime mold, Dictyostelium discoideum could potentially be performed in the flight verified hardware of the EMCS ARC Seed Cassettes.

biocompatibility↗

Cellular Response to Bleomycin-Induced DNA Damage in Human Fibroblast Cells in Space

Living organisms are constantly exposed to space radiation that consists of energetic protons and other heavier charged particles. Whether spaceflight factors, microgravity in particular, affects on the cellular response to DNA damage induced by exposures to radiation or other toxic chemicals will have an impact on the radiation risks for the astronauts, as well as on the mutation rate in microorganisms, is still an open question. Although the possible synergistic effects of space radiation and other spaceflight factors have been investigated since the early days of the human space program, the published results were mostly conflicting and inconsistent. To investigate the effects of spaceflight on the cellular response to DNA damages, human fibroblast cells flown to the International Space Station (ISS) were treated with bleomycin for three hours in the true microgravity environment, which induces DNA damages including the double strand breaks (DSB) similar to the ionizing radiation. Damage in the DNA was measured by the phosphorylation of a histone protein H2AX (-H2AX), which showed slightly more foci in the cells on ISS than in the ground control. The expression of genes involved in the DNA damage response was also analyzed using the PCR array. Although a number of the genes, including CDKN1A and PCNA, were significantly altered in the cells after bleomycin treatment, no significant difference in the expression profile of DNA damage response genes was found between the flight and ground samples. At the time of the bleomycin treatment, the cells on the ISS were found to be proliferating faster than the ground control as measured by the percentage of cells containing positive Ti-67 signals. Our results suggested that the difference in -H2AX between flight and ground was due to the faster growth rate of the cells in space, but spaceflight did not affect the response of the DNA damage response genes to bleomycin treatment.

Lu, Tao↗

Cellular Response to Bleomycin-Induced DNA Damage in Human Fibroblast Cells in Space

Outside the protection of the geomagnetic field, astronauts and other living organisms are constantly exposed to space radiation that consists of energetic protons and other heavier charged particles. Whether spaceflight factors, microgravity in particular, have effects on cellular responses to DNA damage induced by exposure to radiation or cytotoxic chemicals is still unknown, as is their impact on the radiation risks for astronauts and on the mutation rate in microorganisms. Although possible synergistic effects of space radiation and other spaceflight factors have been investigated since the early days of the human space program, the published results were mostly conflicting and inconsistent. To investigate effects of spaceflight on cellular responses to DNA damages, human fibroblast cells flown to the International Space Station (ISS) were treated with bleomycin for three hours in the true microgravity environment, which induced DNA damages including double-strand breaks (DSB) similar to the ionizing radiation. Damages in the DNA were measured by the phosphorylation of a histone protein H2AX (g-H2AX), which showed slightly more foci in the cells on ISS than in the ground control. The expression of genes involved in DNA damage response was also analyzed using the PCR array. Although a number of the genes, including CDKN1A and PCNA, were significantly altered in the cells after bleomycin treatment, no significant difference in the expression profile of DNA damage response genes was found between the flight and ground samples. At the time of the bleomycin treatment, the cells on the ISS were found to be proliferating faster than the ground control as measured by the percentage of cells containing positive Ki-67 signals. Our results suggested that the difference in g-H2AX focus counts between flight and ground was due to the faster growth rate of the cells in space, but spaceflight did not affect initial transcriptional responses of the DNA damage response genes to bleomycin treatment.

Lu, Tao↗

Biomolecular Analysis Capability for Cellular and Omics Research on the International Space Station

International Space Station (ISS) assembly complete ushered a new era focused on utilization of this state-of-the-art orbiting laboratory to advance science and technology research in a wide array of disciplines, with benefits to Earth and space exploration. ISS enabling capability for research in cellular and molecular biology includes equipment for in situ, on-orbit analysis of biomolecules. Applications of this growing capability range from biomedicine and biotechnology to the emerging field of Omics. For example, Biomolecule Sequencer is a space-based miniature DNA sequencer that provides nucleotide sequence data for entire samples, which may be used for purposes such as microorganism identification and astrobiology. It complements the use of WetLab-2 SmartCycler"TradeMark", which extracts RNA and provides real-time quantitative gene expression data analysis from biospecimens sampled or cultured onboard the ISS, for downlink to ground investigators, with applications ranging from clinical tissue evaluation to multigenerational assessment of organismal alterations. And the Genes in Space-1 investigation, aimed at examining epigenetic changes, employs polymerase chain reaction to detect immune system alterations. In addition, an increasing assortment of tools to visualize the subcellular distribution of tagged macromolecules is becoming available onboard the ISS. For instance, the NASA LMM (Light Microscopy Module) is a flexible light microscopy imaging facility that enables imaging of physical and biological microscopic phenomena in microgravity. Another light microscopy system modified for use in space to image life sciences payloads is initially used by the Heart Cells investigation ("Effects of Microgravity on Stem Cell-Derived Cardiomyocytes for Human Cardiovascular Disease Modeling and Drug Discovery"). Also, the JAXA Microscope system can perform remotely controllable light, phase-contrast, and fluorescent observations. And upcoming confocal microscopy capability will allow for optical sectioning of biological tissues to determine microanatomical localization of biomarkers. Furthermore, NASA's geneLAB effort addresses integration of genomic, epigenomic, transcriptomic, proteomic and metabolomic datasets, by applying an innovative open source science platform for multi-investigator high throughput utilization of the ISS. In sum, the expanding ISS capability for analysis of biomolecules is enabling innovative research in a broad spectrum of areas such as cellular and molecular biology, biotechnology, tissue engineering, biomedicine, and Omics, providing manifold benefits for humanity.

Guinart-Ramirez, Y.↗

Translational Cellular Research on the International Space Station

The emerging field of Translational Research aims to coalesce interdisciplinary findings from basic science for biomedical applications. To complement spaceflight research using human subjects, translational studies can be designed to address aspects of space-related human health risks and help develop countermeasures to prevent or mitigate them, with therapeutical benefits for analogous conditions experienced on Earth. Translational research with cells and model organisms is being conducted onboard the International Space Station (ISS) in connection with various human systems impacted by spaceflight, such as the cardiovascular, musculoskeletal, and immune systems. Examples of recent cell-based translational investigations on the ISS include the following. The JAXA investigation Cell Mechanosensing seeks to identify gravity sensors in skeletal muscle cells to develop muscle atrophy countermeasures by analyzing tension fluctuations in the plasma membrane, which changes the expression of key proteins and genes. Earth applications of this study include therapeutic approaches for some forms of muscular dystrophy, which appear to parallel aspects of muscle wasting in space. Spheroids is an ESA investigation examining the system of endothelial cells lining the inner surface of all blood vessels in terms of vessel formation, cellular proliferation, and programmed cell death, because injury to the endothelium has been implicated as underpinning various cardiovascular and musculoskeletal problems arising during spaceflight. Since endothelial cells are involved in the functional integrity of the vascular wall, this research has applications to Earth diseases such as atherosclerosis, diabetes, and hypertension. The goal of the T-Cell Activation in Aging NASA investigation is to understand human immune system depression in microgravity by identifying gene expression patterns of candidate molecular regulators, which will provide further insight into factors that may play a critical role in immune function loss during aging. In addition, Omics investigations with cells have synergistic applications ranging from the evaluation of pharmacological countermeasures to drug discovery. Thus, cell-based translational research onboard the ISS is bidirectionally bridging cutting-edge cellular and molecular approaches with space bioastronautics and human health methodologies on Earth.

Love, John↗

The Role of CDKN1a/p21 in Cellular Senescence of Bone Marrow Stem Cells Under Spaceflight Stressors

Spaceflight environments and their associated conditions, such as microgravity and space radiation, cause many biological functions formerly considered to be standard to behave in nonstandard ways. Exposure to microgravity has shown to induce deleterious effects in stem cell-based tissue regeneration, leading to immune system and healing response impairments as well as muscle and bone density loss. Such risks must be mitigated in order for long-term human space exploration to proceed. Thus, our work seeks to explore mechanisms of stem cell-based tissue regeneration that experience changes in spaceflight environments. Cellular senescence is a process of inducing cell cycle arrest that can be initiated by various stimuli. This function is influenced by two major pathways, controlled by p53 and pRB tumor suppressor proteins. p53 activity targets the cyclin-dependent kinase inhibitor gene p21Cdkn1a in osteogenic cell cycle arrest. Under conditions of mechanical unloading, stem cell-based tissue regeneration has shown to be decreased in both proliferation and differentiation, as many cells are arrested in progenitor states. p21 has shown upregulation in expression under conditions of microgravity, suggesting its role in regenerative bone formation arrest in space. p21 levels are found to be elevated independent of p53, suggesting a decrease in proliferation and regeneration without apoptosis, but rather through cell cycle arrest alone. Thus, we hypothesize that p21 is a mediator of cellular senescence in bone marrow stem cells. Culturing of bone marrow stem cells from wild type and p21 knockout mice under osteoblastogenic conditions will be completed to explore the role of p21Cdkn1a in stem cell proliferation and maturation. We believe that decreases in somatic stem cell differentiation may occur after spaceflight due to signal pathway alterations that result in downstream inhibition of genes involved in differentiation, preventing tissue from repairing and regenerating normally.

Stem Cells↗

A general kinetic framework for dislocation mobility derived from probabilistic cellular automaton simulations of the kink-pair mechanism

Dislocation mobility laws are essential components of dislocation-density-based crystal plasticity models. For dislocations governed by the kink-pair mechanism, however, existing formulations are typically limited to specific regimes due to the com plex interplay between stochastic kink-pair nucleation and lateral kink migration. In this work, we develop a general kinetic framework that expresses the average dislocation velocity as a function of mechanism-level variables: positive/negative kink pair nucleation rates, kink migration velocity, dislocation segment length, critical kink-pair width, and kink height. Probabilis tic cellular automaton simulations are used to capture the behavior of conceptual dislocation segments between the limiting conditions of migration outpacing nucleation on the one end and nucleation outpacing migration on the other. An elemen tary functional form that captures the system dynamics is suggested and fitted against the simulation results. This framework remains valid for arbitrary combinations of the six variables and is, therefore, compatible with any admissible constitutive re lations that describe their stress and temperature dependence. Comparisons with established approaches and experimental results confirm the robustness and physical consistency of the formulation, making it broadly applicable to material systems in which dislocation motion is governed by the kink-pair mechanism.

36 MATERIALS SCIENCE↗

Insight into Molecular Basis and Dynamics of Full-length CRaf Kinase in Cellular Signaling Mechanism

This study presents the first large-scale simulation using an initial structure predicted by AI/ML algorithms for the 648-amino-acid CRaf kinase, which plays a key role in cellular signaling. Simulation results show the evolution of the predicted structure into much more compact structures with inter-domain interactions that shed insights into auto-inhibition mechanism, paradoxical effect, activation, and recruitment pathways in the CRaf kinase. Newly identified epitopes in the CRaf may suggest additional drug targets. The results were published in Biophysical Journal, DOI:10.1016/j.bpj.2024.06.028.

59 BASIC BIOLOGICAL SCIENCES↗

Integration of Mobil Satellite and Cellular Systems

By integrating the ground based infrastructure component of a mobile satellite system with the infrastructure systems of terrestrial 800 MHz cellular service providers, a seamless network of universal coverage can be established.

cellular satellite Mobile Telephone Switching Offi↗

On the numerical sensitivity of cellular automata grain structure predictions to large thermal gradients and cooling rates

Cellular automata (CA) models of as-solidified grain structure, originally developed and applied to casting, have become a common means of predicting grain structure resulting from Additive Manufacturing (AM) processes. The majority of these models are based on the decentered octahedron approach, which attempts to correct for the effect of grid anisotropy on the prediction of competitive solidification of dendritic grains. However, AM solidification occurs under cooling rates ($\dot{T}$) and thermal gradients (G) that are orders of magnitude larger than those encountered in casting, and no systematic investigation on the effect of the CA model cell size (Δx) and time step (Δt) on AM microstructure predictions has been performed. Here, in this study, such an investigation is first performed via simulation of individual grains of various crystallographic orientations with a fixed, unidirectional G, showing that CA prediction of the steady-state undercooling matched the expected values based on the interfacial response function at small G and deviated from the expected values at large G. Simulation of competitive growth of multiple grains showed a weakening of the predicted texture as G and Δx became large. Simulation of solidification under AM conditions, where G and $\dot{T}$ vary spatially across the melt pools, showed that not only does grain selection weaken and deviate from expectations at large Δx, but grains with crystallographic $\langle$100$\rangle$ aligned with the grid directions are more adversely affected by the temperature field discontinuities than grains with other crystallographic orientations. Despite the fact that the exact grain competition results depended on Δt, the overall texture development was notably less sensitive to Δt than Δx, provided that a reasonable value of Δt is selected based on the ratio of Δx to the maximum local solidification velocity in the simulation domain. Finally, from the directional solidification and AM simulation results, an analysis of computational cost compared to simulation resolution is performed based on an equation derived to quantify the relatively inaccuracy in grain selection based on the model and temperature field inputs. From this analysis, it is concluded that there is a need for algorithmic improvements to improve CA grain competition accuracy for large G processing conditions as sufficiently small Δx to resolve the necessary competition is intractable for many AM processing conditions.

36 MATERIALS SCIENCE↗

The effect of varying powder feedstock chemistry and printing atmosphere on the microstructure of additively manufactured nickel-based ODS alloys: Role on stabilization of cellular structures vs. oxide dispersion formation

Nickel-based alloys have a wide variety of structural applications due to their high corrosion resistance and mechanical strength which depend on solid solution strengthening, or the formation of oxides and/or intermetallic precipitation for their properties. In this study, the microstructure of six Ni–Cr–Y–Ti–Al powder batches designed for the production of oxide dispersion strengthened nickel were compared. These batches varied in chemistry and atomization technique used which included Gas Atomization Reactive Synthesis (GARS). The batches of powder were then consolidated via Additive Manufacturing (AM) Powder Bed Fusion using Laser Beam (PBF-LB) and characterized via transmission electron microscopy to elucidate the influence of powder feedstock (i.e. synthesis methodology and chemistry) on the PBF-LB microstructure. The study investigates (i) how the amount of yttrium and titanium additions in the powder feedstock and the addition of oxygen during the processing (through GARS) affect the microstructure of the powder itself and the AM printed microstructure, and (ii) how the control of oxygen addition in the printing atmosphere during the PBF-LB printing process itself is another important parameter for achieving the formation of the wanted oxide dispersion versus the stabilization of the cellular structure (often observed in AM processed alloys). Microstructural characterization of both powder particles and additively manufactured nickel alloys in this study provide important insights into the movement of yttrium within the material upon solidification, particularly along cell boundaries, and how yttrium behaves depending on alloy chemistry. When a threshold of yttrium content is reached within the system, yttrium consistently reacts to form an intermetallic along cell boundaries instead of forming oxide nanoparticles.

36 MATERIALS SCIENCE↗