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Final Report - Hypergravity Effects on Chromatin Conformation and Nuclear Structure in Cultured Cells using High-throughput Sequencing

The conformation of the human genome is known to play an important role in transcriptional control of gene expression. Our aim is to assess whether exposure of cultured human cardiomyocytes to hypergravity can induce changes in chromatin organization as assayed by Hi-C proximity ligation technique. Hi-C is used to analyze chromatin interactions by using formaldehyde to crosslink regions of chromatin that are in close proximity. The DNA is then fragmented using a restriction enzyme and ligated under dilute conditions to favor intramolecular ligation of cross-linked fragments. Finally, the DNA is sequenced, allowing reconstruction of genomic structure. iCell cardiomyocyte cultures will be exposed to 40 g via an engineered device (incu-fuge) that allows for the chemical fixation of cells on a spinning centrifuge. Additionally, we plan on using fluorescence microscopy to analyze the nuclear and actin cytoskeletal conformation of cardiomyocytes post hypergravity exposure. We hypothesize that chromatin will rearrange in hypergravity conditions, both rapidly due to direct mechanical forces, as well as over longer time frames due to changes to structures necessary for transcriptional responses such as the de novo formation of promoter-enhancer loops. A negative result–no change between 1 g and hypergravity conditions–would nevertheless be an important data point in our understanding of the rheology of the nucleus, and how cells and cellular structures respond to different gravity fields. The cellular physiology of hypergravity has clinical relevance for spaceflight, and can further inform our understanding of microgravity physiology.

chromatin↗

Numerical simulation of tornado wind loading on structures

A numerical simulation of a tornado interacting with a building was undertaken in order to compare the pressures due to a rotational unsteady wind with that due to steady straight winds used in design of nuclear facilities. The numerical simulations were performed on a two-dimensional compressible hydrodynamics code. Calculated pressure profiles for a typical building were then subjected to a tornado wind field and the results were compared with current quasisteady design calculations. The analysis indicates that current design practices are conservative.

Maiden, D. E.↗

Fundamental space radiobiology

The unique feature of the space radiation environment is the dominance of high-energy charged particles (HZE or high LET radiation) emitted by the Sun and galactic sources, or trapped in the Van Allen radiation belts. These charged particles present a significant hazard to space flight crews, and accelerator-based experiments are underway to quantify the health risks due to unavoidable radiation exposure. There are three principal properties of charged particles that distinguish them from conventional radiation, i.e. gamma rays and x-rays. First, they have a defined range in matter rather than an exponential absorption profile. Second, they undergo nuclear reactions to produce secondary particles. Third, and most important, they deposit their energy along well-defined linear paths or tracks rather than diffuse fields. The structured energy deposition pattern interacts on multiple scales with the biological structures of DNA, cells and tissues to produce correlated patterns of damage that evade repair systems. Traditional concepts of dose and its associated normalization parameter, RBE (relative biological effectiveness), break down under experimental scrutiny, and probabilistic models of risk based on the number of particle traversals per cell may be more appropriate. Unique patterns of DNA damage, gene expression, mobilization of repair proteins, activation of cytokines and remodeling of cellular microenvironment are observed following exposure to high LET radiation. At low levels of exposure the communication of bioactive substances from irradiated to unirradiated "bystander" cells can amplify the damage and cause a significant deviation from linearity in dose vs. response relations. Under some circumstances, there is even a multigenerational delay in the expression of radiation-induced genetic damage (genomic instability) which is not strictly dose dependent. These issues and the experimental evidence derived from ground based experiments at particle accelerators are presented along with speculation about how modified inertial conditions might perturb homeostatic responses to radiation to further complicate risk assessment for space flight.

NASA Discipline Radiation Health↗

The Host Galaxies of X-Ray Selected Active Galactic Nuclei to z - 2.5: Structure, Star-Formation and Their Relationships from CANDELS and Herschel/Pacs

We study the relationship between the structure and star-formation rate (SFR) of X-ray selected low and moderate luminosity active galactic nuclei (AGNs) in the two Chandra Deep Fields, using Hubble Space Telescope imaging from the Cosmic Assembly Near Infrared Extragalactic Legacy Survey (CANDELS) and deep far-infrared maps from the PEP+GOODS-Herschel survey. We derive detailed distributions of structural parameters and FIR luminosities from carefully constructed control samples of galaxies, which we then compare to those of the AGNs. At z is approximately 1, AGNs show slightly diskier light profiles than massive inactive (non-AGN) galaxies, as well as modestly higher levels of gross galaxy disturbance (as measured by visual signatures of interactions and clumpy structure). In contrast, at z ∼ 2, AGNs show similar levels of galaxy disturbance as inactive galaxies, but display a red central light enhancement, which may arise due to a more pronounced bulge in AGN hosts or due to extinguished nuclear light. We undertake a number of tests of both these alternatives, but our results do not strongly favour one interpretation over the other. The mean SFR and its distribution among AGNs and inactive galaxies are similar at z greater than 1.5. At z less than 1, however, clear and significant enhancements are seen in the SFRs of AGNs with bulge-dominated light profiles. These trends suggest an evolution in the relation between nuclear activity and host properties with redshift towards a minor role for mergers and interactions at z greater than 15

host↗

Mechanotransduction and the functional response of bone to mechanical strain

Mechanotransduction plays a crucial role in the physiology of many tissues including bone. Mechanical loading can inhibit bone resorption and increase bone formation in vivo. In bone, the process of mechanotransduction can be divided into four distinct steps: (1) mechanocoupling, (2) biochemical coupling, (3) transmission of signal, and (4) effector cell response. In mechanocoupling, mechanical loads in vivo cause deformations in bone that stretch bone cells within and lining the bone matrix and create fluid movement within the canaliculae of bone. Dynamic loading, which is associated with extracellular fluid flow and the creation of streaming potentials within bone, is most effective for stimulating new bone formation in vivo. Bone cells in vitro are stimulated to produce second messengers when exposed to fluid flow or mechanical stretch. In biochemical coupling, the possible mechanisms for the coupling of cell-level mechanical signals into intracellular biochemical signals include force transduction through the integrin-cytoskeleton-nuclear matrix structure, stretch-activated cation channels within the cell membrane, G protein-dependent pathways, and linkage between the cytoskeleton and the phospholipase C or phospholipase A pathways. The tight interaction of each of these pathways would suggest that the entire cell is a mechanosensor and there are many different pathways available for the transduction of a mechanical signal. In the transmission of signal, osteoblasts, osteocytes, and bone lining cells may act as sensors of mechanical signals and may communicate the signal through cell processes connected by gap junctions. These cells also produce paracrine factors that may signal osteoprogenitors to differentiate into osteoblasts and attach to the bone surface. Insulin-like growth factors and prostaglandins are possible candidates for intermediaries in signal transduction. In the effector cell response, the effects of mechanical loading are dependent upon the magnitude, duration, and rate of the applied load. Longer duration, lower amplitude loading has the same effect on bone formation as loads with short duration and high amplitude. Loading must be cyclic to stimulate new bone formation. Aging greatly reduces the osteogenic effects of mechanical loading in vivo. Also, some hormones may interact with local mechanical signals to change the sensitivity of the sensor or effector cells to mechanical load.

Non-NASA Center↗

High temperature transport properties of air

A general computer code was developed to allow calculation of atom-atom and ion-atom transport collision integrals from accurate potential energy curves described by a set of discrete data points for a broad range of scattering conditions. This code is based upon semiclassical approximations that properly account for quantum mechanical behavior such as tunneling effects near a barrier maximum, resonance charge exchange, and nuclear symmetry effects. Transport collision integrals were determined for N-N, O-O, N(+)-N, and O(+)-O interactions from complete sets of accurate potential functions derived from combined experimental and ab initio structure calculations. For the O-O case, this includes results for excited states. The calculated values of the N(+)-N and O(+)-O resonance charge exchange cross section Q(ex) agree well with measurements from beam experiment that are available at high energies where the diffusion cross section Q(d) satisfies Q(d) approximately equal to 2Q(ex).

Levin, E.↗

In situ chemical analyses of extraterrestrial bodies

One of the most important tasks on any sample return mission will have to be a quick sample characterization in order to guarantee a variety of collected samples. An alpha particle instrument with alpha, proton and X-ray modes can provide a quick and almost complete chemical analysis of Mars samples. This instrument is based on three interactions of the alpha particles from a radioactive source with matter: elastic scattering of the alpha particles by nuclei (alpha mode), (alpha,p) nuclear reaction with some light elements (proton mode), and excitation of the atomic structure of atoms by alpha particles, leading to emission of characteristic X-rays of the lunar surface at three sites during the Surveyor mission of 1967 to 1968. Since then the instrument has been improved and miniaturized substantially. As shown in the past, the alpha particle instrument can operate under Martian conditions without any degradation in the performance. The alpha and proton modes can provide vital information about the light elements, while the X-ray mode with its ambient temperature X-ray detector will be useful for the heavier elements. The excitation of the atomic structure is provided by the same alpha radioactive source that is used by alpha and proton modes or by an auxiliary X-ray source that is selected to enhance the sensitivity to some important geochemical elements.

Economou, Thanasis E.↗

Micronuclei Induction in Human Fibroblasts Exposed In Vitro to Los Alamos High-Energy Neutrons

High-energy secondary neutrons, produced by the interaction of galactic cosmic rays with the atmosphere, spacecraft structure and planetary surfaces, contribute to a significant fraction to the dose equivalent in crew members and passengers during commercial aviation travel, and astronauts in space missions. The Los Alamos Nuclear Science Center (LANSCE) neutron facility#s ICE House 30L beamline is known to generate neutrons that simulate the secondary neutron spectra of earth#s atmosphere. The neutron spectrum is also similar to that measured onboard spacecraft like the MIR and International Space Station (ISS). To evaluate the biological damage, we exposed human fibroblasts in vitro to the LANSCE neutron beams without degrader at an entrance dose rate of 25 mGy/hr and analyzed the micronuclei (MN) induction. The cells were also placed behind a 9.9 cm water column to study effect of shielding in the protection of neutron induced damages. It was found that the dose response in the MN frequency was linear for the samples with and without shielding and the slope of the MN yield behind the shielding was reduced by a factor of 3.5. Compared to the MN induction in human fibroblasts exposed to a gamma source at a low dose rate, the RBE was found to be 16.7 and 10.0 for the neutrons without and with 9.9 cm water shielding, respectively.

Gersey, Brad↗

M-BAND Analysis of Chromosome Aberration In Human Epithelial Cells exposed to Gamma-ray and Secondary Neutrons of Low Dose Rate

High-energy secondary neutrons, produced by the interaction of galactic cosmic rays with the atmosphere, spacecraft structure and planetary surfaces, contribute to a significant fraction to the dose equivalent in crew members and passengers during commercial aviation travel, and astronauts in space missions. The Los Alamos Nuclear Science Center (LANSCE) neutron facility's "30L" beam line is known to generate neutrons that simulate the secondary neutron spectrum of the Earth's atmosphere at high altitude. The neutron spectrum is also similar to that measured onboard spacecraft like the MIR and the International Space Station (ISS). To evaluate the biological damage, we exposed human epithelial cells in vitro to the LANSCE neutron beams at an entrance dose rate of 2.5 cGy/hr or gamma-ray at 1.7cGy/hr, and assessed the induction of chromosome aberrations that were identified with mBAND. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of inter-chromosomal aberrations (translocation to unpainted chromosomes) and intra-chromosomal aberrations (inversions and deletions within a single painted chromosome). Compared to our previous results for gamma-rays and 600 MeV/nucleon Fe ions of high dose rate, the neutron data showed a higher frequency of chromosome aberrations. However, detailed analysis of the inversion type revealed that all of the three radiation types in the study induced a low incidence of simple inversions. The low dose rate gamma-rays induced a lower frequency of chromosome aberrations than high dose rate gamma-rays, but the inversion spectrum was similar for the same cytotoxic effect. The distribution of damage sites on chromosome 3 for different radiation types will also be discussed.

Hada, M.↗

Gamma ray astronomy

The various source mechanisms for celestial gamma rays are reviewed. The gamma-ray data are examined as a source of information about the processes and source locations for the production of charged particle cosmic rays, galactic structure, explosive nucleosynthesis in supernovae, regions of confinement for cosmic rays, regions where matter-antimatter annihilation occurs, and the general condition in cosmological space both in the past and present. Topics include gamma rays from pi mesons by nuclear interactions, nuclear and supernovae lines, diffuse emission and discrete sources, interstellar absorption and detection of gamma rays, and others. A brief view of the available gamma-ray detection systems and techniques is presented.

Fichtel, C. E.↗

The diffuse galactic gamma radiation - The Compton contribution and component separation by energy interval and galactic coordinates

The diffuse high-energy galactic gamma radiation to be expected from cosmic ray interactions with matter and photons is considered with particular emphasis on the contribution of Compton radiation from cosmic ray electrons. The intensity, spectrum and spatial distribution of the expected galactic gamma radiation are estimated based on models of the matter, cosmic ray and photon distributions to take into account the contributions of bremsstrahlung, high-energy cosmic-ray nucleon and interstellar matter interactions as well as Compton interactions between cosmic ray electrons and background photons. Results suggest that the Compton gamma ray contribution from cosmic ray electron interactions with galactic visible and infrared photons is substantially larger than previously believed. Analysis of the energy spectra and latitude dependence of the various sources reveals that the Compton radiation, bremsstrahlung and nuclear cosmic ray-matter interaction radiation should be separable, with Compton radiation dominating at energies from 10 to 100 MeV at galactic latitudes greater than several degrees. Results demonstrate the potential of gamma ray observations in studies of galactic structure, cosmic ray electrons and galactic photon density.

Kniffen, D. A.↗

Phenomenological model of nuclear primary air showers

The development of proton primary air showers is described in terms of a model based on a hadron core plus an electromagnetic cascade. The muon component is neglected. The model uses three parameters: a rate at which hadron core energy is converted into electromagnetic cascade energy and a two-parameter sea-level shower-age function. By assuming an interaction length for the primary nucleus, the model is extended to nuclear primaries. Both models are applied over the energy range from 10 to the 13th power to 10 to the 21st power eV. Both models describe the size and age structure (neglecting muons) from a depth of 342 to 2052 g/sq cm.

Tompkins, D. R., Jr.↗

Mitigating Space Radiation Using Magnesium(-Lithium) and Boron Carbide Composites

The health effects of galactic cosmic radiation are a serious impediment to crewed exploration of the solar system. OLTARIS, an interface for the 3DHZETRN deterministic radiation transport code, was used to assess the response of aerospace materials to this constant radiation exposure. Traditional aerospace structural materials like aluminum can, after a certain mass, increase the health effects of such radiation. However, materials with lower atomic mass may mitigate this build-up in secondary radiation with increasing areal density. As such, lower atomic mass structural alloys of magnesium and magnesium–lithium are promising candidates. These alloys may reduce the mass of structures when substituted for aluminum alloys. Reinforcement with boron carbide could further reduce atomic mass while also improving the mechanical properties of such lightweight alloys. This study found that the lower atomic mass of these materials increased nuclear fragmentation upon cosmic radiation interactions, leading to a softening of the secondary (neutron) radiation spectra. This softened spectra reduced the effective dose equivalent, a measure of health effects, for magnesium(-lithium) alloys and their boron carbide-reinforced composites when compared to aluminum.

Galactic cosmic radiation↗

The CO-12 and CO-13 J=2-1 and J=1-0 observations of hot and cold galaxies

Researchers observed the nuclear regions of the galaxies NGC 2146 and IC 342 in CO-12 and CO-13 J=1-0 and J=2-1 lines using the Five College Radio Astronomy Observatory (FCRAO) 14m telescope. NGC 2146 is a peculiar Sab spiral galaxy. Its complex optical morphology and strong nuclear radio continuum emission suggest that it is experiencing a phase of violent activity and could have a polar ring which may have resulted from an interaction. IC 342 is a nearby luminous Scd spiral galaxy. Strong CO, infrared and radio continuum emission from the nuclear region of IC 342 indicate enhanced star-forming activity, and interferometric CO-12 J=1-0 observations reveal a bar-like structure centered on the nucleus, along the dark lane in the NS direction. These two galaxies are selected based on their different dust temperatures and star formation efficiencies (SFE) as derived from the Infrared Astronomy Satellite (IRAS) S sub 60 mu/S sub 100 mu flux density ratio and L sub IR/M(H2), respectively, with a relatively high SFE and dust temperature of 45 K in NGC 2146 and a relatively low SFE and dust temperature of 35 K in IC 342. The data from the different CO-12 and CO-13 lines are used to study the physical conditions in the molecular clouds in the galaxies. Researchers also consider the radiative transfer to determine whether a warm and optically thin gas component exists in these galaxies, as has been suggested in the case of M82 (Knapp et al. 1980), and whether the warm gas is related to the dust properties. Since optically thin CO-12 gas is rarely detected in our own Galaxy (except in outflow sources), to confirm its existence in external galaxies is very important in understanding the molecular content of external galaxies and its relationship to star formation activity. The present CO-12 J=2-1 and CO-13 J=2-1 and J=1-0 data for NGC 2146 are the first detections of this galaxy to our knowledge. The CO-12 J=1-0 distribution in NGC 2146 has been measured as part of the FCRAO Extragalactic Survey. For the well-studied IC 342, the data are compared with 30m observations and other available data. Researchers present the observed results.

Xie, Shuding↗

Heavy-ion fragmentation studies in thick water absorbers

Proposed long-term space missions could expose crewmembers to significant fluxes of galactic cosmic radiation (GCR) particles and secondary particles created from nuclear collisions. An assessment of radiobiological risks is dependent upon an accurate description of the charged-particle radiation field inside the human body. As shield thickness increases and the incident ions are slowed, the production of secondary particles contributes an increasingly significant fraction of the total dose until eventually secondary particles become more important than the primary particles. The nuclear mean free path of the GCR ions (which usually have nuclear charge between 1 (protons) and 26 (iron), both inclusive) are comparable with thicknesses typical of spacecraft structures and the human body. Collisions in these media will create projectile and target fragments with charge less than that of the primary particle, and each interaction event can have a multiplicity of more than one emerging interaction product. Projectile fragments usually continue on with very nearly the velocity of the primary ion (the so-called straightahead approximation). Having sufficient energy, the fragments may collide with atomic nuclei in thick shields and create a second generation of fragments, and so on. Target fragments are emitted from a struck nucleus, usually with much lower energy than projectile fragments and nearly isotropically in the rest frame of the absorbing medium. The resulting spectrum of particles and their energy loss rates will be very different from that in the unshielded environment, will determine the radiobiological impact on exposed living tissues -- whether in space or in ground-based radiobiology experiments -- and will play an important role in radiation effects on microelectronics.

Shavers, M. R.↗

Chromosome Aberrations in Human Epithelial Cells Exposed Los Alamos High-Energy Secondary Neutrons: M-BAND Analysis

High-energy secondary neutrons, produced by the interaction of galactic cosmic rays (GCR) with the atmosphere, spacecraft structure and planetary surfaces, contribute a significant fraction to the dose equivalent radiation measurement in crew members and passengers of commercial aviation travel as well as astronauts in space missions. The Los Alamos Nuclear Science Center (LANSCE) neutron facility's 30L beam line (4FP30L-A/ICE House) is known to generate neutrons that simulate the secondary neutron spectrum of the Earth's atmosphere at high altitude. The neutron spectrum is also similar to that measured onboard spacecrafts like the MIR and the International Space Station (ISS). To evaluate the biological damage, we exposed human epithelial cells in vitro to the LANSCE neutron beams with an entrance dose rate of 2.5 cGy/hr, and studied the induction of chromosome aberrations that were identified with multicolor-banding in situ hybridization (mBAND) technique. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of inter-chromosomal aberrations (translocation to unpainted chromosomes) and intra-chromosomal aberrations (inversions and deletions within a single painted chromosome). Compared to our previous results with gamma-rays and 600 MeV/nucleon Fe ions of high dose rate at NSRL (NASA Space Radiation Laboratory at Brookhaven National Laboratory), the neutron data from the LANSCE experiments showed significantly higher frequency of chromosome aberrations. However, detailed analysis of the inversion type revealed that all of the three radiation types in the study induced a low incidence of simple inversions. Most of the inversions in gamma-ray irradiated samples were accompanied by other types of intrachromosomal aberrations but few inversions were accompanied by interchromosomal aberrations. In contrast, neutrons and Fe ions induced a significant fraction of inversions that involved complex rearrangements of both inter- and intrachromosome exchanges. The distribution of damage sites on chromosome 3 was also compared for different radiation types. The breakpoints were randomly localized on chromosome 3 with neutrons and Fe ions exposure, whereas non-random distribution with clustering breakpoints was observed with gamma-rays exposure. The specific fingerprint of neutron radiations on chromosomal aberrations will be discussed.

Hada, M.↗

The dynamic interstellar medium in NGC 4438

We present multiwavelength observations of the interstellar medium in the disturbed galaxy NGC 4438. We find extended emission-line filaments, which are kinematically unrelated to the galaxy disk. For one of these features, an extended loop, we propose a scheme in which the 'bubble' seen at radio wavelengths has swept a clear path for outflowing material from the nucleus to ionize more distant material. Another radial emission-line feature might represent shocks in the disk along a gravitationally-defined caustic produced during a tidal encounter, or a 'searchlight beam' escaping the inner dusty regions of the galaxy. Energy-balance arguments favor the shock picture, but further study will be important in understanding this feature. Emission-line ratios indicate that shocks may be important in the energetics of the interstellar medium (ISM) through much of the disk of NGC 4438; the alternative is star formation in a uniformly very dense ISM. We use K-band data to locate the stellar peak of the galaxy, which is just outside the prominent radio-continuum loop or shell feature. The nucleus itself is at most a very weak radio source. However, the more extended radio structure shows that energetic events have taken place in the innermost few hundred parsees of NGC 4438. The interstellar medium of NGC 4438 may show at once the impact of nuclear energy input (from an AGN or cocooned starburst), perturbation by the companion NGC 4435, and interaction with the dense intergalactic medium in the Virgo core.

Keel, William C.↗

Cosmic ray secondary nuclei and the structure of the galaxy

The consequencies of diffusive acceleration of cosmic rays in supernova shocks propagation through an inhomogeneous interstellar medium are explored. The acceleration takes place in the hot, tenuous, intercloud gas, while nuclear collisions, leading to the production of cosmic ray secondaries, predominantly occur in those regions where the supernova shocks collide with interstellar clouds. A simple model is used to calculate the interaction of a (cosmic ray + gas) shock with a cloud, and thus determine the gross topology. Extending this to the whole system, using mean cloud sizes and space densities, allows us to calculate the secondary/primary cosmic ray abundance ratios for light and heavy nuclei.

Morfill, G. E.↗