Evaluation of surgical skills in microgravity using force sensing.

INTRODUCTION: Force measurements can be used to characterize surgical maneuvers in microgravity. METHODS: : A series of surgical tasks was performed by a group of 20 participants (n=20) both in 1g on the ground and in 0 g aboard NASA's KC-135 aircraft in parabolic flight. The group included astronauts, a flight surgeon, surgeons, physicians, Ph.D.-scientists, and technical personnel. The interaction forces between the surgical instruments and the mock tissue were measured for a clip-applying, suturing, grasping, and cutting. Seven evaluations in 1g and a single evaluation in 0 g were performed by each of the participants. RESULTS: The data from a single participant are examined in detail. Statistical results for the group of 20 participants do not show significant differences in the average or peak forces during clip-applying or in the average forces applied during suturing in 0 g versus in 1g. However, the results do show statistically greater (43% higher) peak forces during suturing in microgravity. DISCUSSION: These data show the usefulness of analyzing force information to assess surgical task performance in microgravity. Although peak suturing forces were statistically higher in microgravity, their clinical relevance is unknown, but likely would not result in a change in clinical outcome. Overall, the data suggest that forces exerted during surgical tasks will not pose a significant barrier to effective surgery in microgravity.

Simulation of gaseous diffusion in partially saturated porous media under variable gravity with lattice Boltzmann methods.

Liquid distributions in unsaturated porous media under different gravitational accelerations and corresponding macroscopic gaseous diffusion coefficients were investigated to enhance understanding of plant growth conditions in microgravity. We used a single-component, multiphase lattice Boltzmann code to simulate liquid configurations in two-dimensional porous media at varying water contents for different gravity conditions and measured gas diffusion through the media using a multicomponent lattice Boltzmann code. The relative diffusion coefficients (D rel) for simulations with and without gravity as functions of air-filled porosity were in good agreement with measured data and established models. We found significant differences in liquid configuration in porous media, leading to reductions in D rel of up to 25% under zero gravity. The study highlights potential applications of the lattice Boltzmann method for rapid and cost-effective evaluation of alternative plant growth media designs under variable gravity.

Physical and hydraulic properties of baked ceramic aggregates used for plant growth medium.

Baked ceramic aggregates (fritted clay, arcillite) have been used for plant research both on the ground and in microgravity. Optimal control of water and air within the root zone in any gravity environment depends on physical and hydraulic properties of the aggregate, which were evaluated for 0.25-1-mm and 1-2-mm particle size distributions. The maximum bulk densities obtained by any packing technique were 0.68 and 0.64 g cm-3 for 0.25-1-mm and 1-2-mm particles, respectively. Wettable porosity obtained by infiltration with water was approximately 65%, substantially lower than total porosity of approximately 74%. Aggregate of both particle sizes exhibited a bimodal pore size distribution consisting of inter-aggregate macropores and intra-aggregate micropores, with the transition from macro- to microporosity beginning at volumetric water content of approximately 36% to 39%. For inter-aggregate water contents that support optimal plant growth there is 45% change in water content that occurs over a relatively small matric suction range of 0-20 cm H2O for 0.25-1-mm and 0 to -10 cm H2O for 1-2-mm aggregate. Hysteresis is substantial between draining and wetting aggregate, which results in as much as a approximately 10% to 20% difference in volumetric water content for a given matric potential. Hydraulic conductivity was approximately an order of magnitude higher for 1-2-mm than for 0.25-1-mm aggregate until significant drainage of the inter-aggregate pore space occurred. The large change in water content for a relatively small change in matric potential suggests that significant differences in water retention may be observed in microgravity as compared to earth.

Cytogenetic effects of high-energy iron ions: dependence on shielding thickness and material.

We report results for chromosomal aberrations in human peripheral blood lymphocytes after they were exposed to high-energy iron ions with or without shielding at the HIMAC, AGS and NSRL accelerators. Isolated lymphocytes were exposed to iron ions with energies between 200 and 5000 MeV/nucleon in the 0.1-1-Gy dose range. Shielding materials consisted of polyethylene, lucite (PMMA), carbon, aluminum and lead, with mass thickness ranging from 2 to 30 g/cm2. After exposure, lymphocytes were stimulated to grow in vitro, and chromosomes were prematurely condensed using a phosphatase inhibitor (calyculin A). Aberrations were scored using FISH painting. The yield of total interchromosomal exchanges (including dicentrics, translocations and complex rearrangements) increased linearly with dose or fluence in the range studied. Shielding decreased the effectiveness per unit dose of iron ions. The highest RBE value was measured with the 1 GeV/nucleon iron-ion beam at NSRL. However, the RBE for the induction of aberrations apparently is not well correlated with the mean LET. When shielding thickness was increased, the frequency of aberrations per particle incident on the shield increased for the 500 MeV/nucleon ions and decreased for the 1 GeV/nucleon ions. Maximum variation at equal mass thickness was obtained with light materials (polyethylene, carbon or PMMA). Variations in the yield of chromosomal aberrations per iron particle incident on the shield follow variations in the dose per incident particle behind the shield but can be modified by the different RBE of the mixed radiation field produced by nuclear fragmentation. The results suggest that shielding design models should be benchmarked using both physics and biological data.

Stability of chromosome aberrations in the blood lymphocytes of astronauts measured after space flight by FISH chromosome painting.

Follow-up measurements of chromosome aberrations in the blood lymphocytes of astronauts were performed by FISH chromosome painting at various intervals from 5 months to more than 5 years after space flight and compared to preflight baseline measurements. For five of the six astronauts studied, the analysis of individual time courses for translocations revealed a temporal decline of yields with half-lives ranging from 10 to 58 months. The yield of exchanges remained unchanged for the sixth astronaut during an observation period of 5 months after flight. These results may indicate complications with the use of stable aberrations for retrospective dose reconstruction, and the differences in the decay time may reflect individual variability in risk from space radiation exposure.

Caffeine sensitizes nondividing human fibroblasts to x rays by inducing a high frequency of misrepair.

Caffeine sensitizes cells to ionizing radiation, and this effect is believed to be associated with the disruption of DNA damage-responsive cell cycle checkpoints, which is controlled by ATM. Recent studies suggest that misrejoining of DSBs is one of the underlying mechanisms of AT cell hyper-radiosensitivity. In this study, we investigated the effects of caffeine and radiation on nongrowing G0 normal human fibroblast cells by determining cell survival and scoring aberrations in calyculin A-induced G2 chromosomes. Results from the cell survival study indicate that after X-ray exposure G0 cells were sensitized by 24 h treatment with caffeine. Analysis of chromosome aberrations using FISH (fluorescence in situ hybridization) revealed a high frequency of aberrant cells and color junctions in the caffeine-treated cells. Since most DNA repair in nongrowing G0 cells is believed to result from nonhomologous end joining (NHEJ), caffeine may influence the fidelity of the NHEJ pathway in irradiated G0 cells.

Immunofluorescence detection of clustered gamma-H2AX foci induced by HZE-particle radiation.

We studied the spatial and temporal distributions of foci of the phosphorylated form of the histone protein H2AX (gamma-H2AX), which is known to be activated by double-strand breaks after irradiation of human fibroblast cells with high-energy silicon (54 keV/microm) and iron (176 keV/microm) ions. Here we present data obtained with the ion path parallel to a monolayer of human fibroblast cells that leads to gamma-H2AX aggregates in the shape of streaks stretching over several micrometers in an x/y plane, thus enabling the analysis of the fluorescence distributions along the ion trajectories. Qualitative analyses of these distributions provide insights into DNA damage processing kinetics for high charge and energy (HZE) ions, including evidence of increased clustering of DNA damage and slower processing with increasing LET.

Measurement of hydraulic characteristics of porous media used to grow plants in microgravity.

Understanding the effect of gravity on hydraulic properties of plant growth medium is essential for growing plants in space. The suitability of existing models to simulate hydraulic properties of porous medium is uncertain due to limited understanding of fundamental mechanisms controlling water and air transport in microgravity. The objective of this research was to characterize saturated and unsaturated hydraulic conductivity (K) of two particle-size distributions of baked ceramic aggregate using direct measurement techniques compatible with microgravity. Steady state (Method A) and instantaneous profile measurement (Method B) methods for K were used in a single experimental unit with horizontal flow through thin sections of porous medium providing an earth-based analog to microgravity. Comparison between methods was conducted using a crossover experimental design compatible with limited resources of space flight. Satiated (natural saturation) K ranged from 0.09 to 0.12 cm s-1 and 0.5 to >1 cm s-1 for 0.25- to 1- and 1- to 2-mm media, respectively. The K at the interaggregate/intraaggregate transition was approximately 10(-4) cm s-1 for both particle-size distributions. Significant differences in log(10)K due to method and porous medium were less than one order of magnitude and were attributed to variability in air entrapment. The van Genuchten/Mualem parametric models provided an adequate prediction of K of the interaggregate pore space, using residual water content for that pore space. The instantaneous profile method covers the range of water contents relevant to plant growth using fewer resources than Method A, all advantages for space flight where mass, volume, and astronaut time are limited.

Discontinuous pore fluid distribution under microgravity--KC-135 flight investigations.

Designing a reliable plant growth system for crop production in space requires the understanding of pore fluid distribution in porous media under microgravity. The objective of this experimental investigation, which was conducted aboard NASA KC-135 reduced gravity flight, is to study possible particle separation and the distribution of discontinuous wetting fluid in porous media under microgravity. KC-135 aircraft provided gravity conditions of 1, 1.8, and 10(-2) g. Glass beads of a known size distribution were used as porous media; and Hexadecane, a petroleum compound immiscible with and lighter than water, was used as wetting fluid at residual saturation. Nitrogen freezer was used to solidify the discontinuous Hexadecane ganglia in glass beads to preserve the ganglia size changes during different gravity conditions, so that the blob-size distributions (BSDs) could be measured after flight. It was concluded from this study that microgravity has little effect on the size distribution of pore fluid blobs corresponding to residual saturation of wetting fluids in porous media. The blobs showed no noticeable breakup or coalescence during microgravity. However, based on the increase in bulk volume of samples due to particle separation under microgravity, groups of particles, within which pore fluid blobs were encapsulated, appeared to have rearranged themselves under microgravity.

Dissolution kinetics of iron-, manganese-, and copper-containing synthetic hydroxyapatites.

Micronutrient-substituted synthetic hydroxyapatite (SHA) is being evaluated by the National Aeronautics and Space Administration's (NASA) Advanced Life Support (ALS) Program for crop production on long-duration human missions to the International Space Station or for future Lunar or Martian outposts. The stirred-flow technique was utilized to characterize Ca, P, Fe, Mn, and Cu release characteristics from Fe-, Mn-, and Cu-containing SHA in deionized (DI) water, citric acid, and diethylene-triamine-pentaacetic acid (DTPA). Initially, Ca and P release rates decreased rapidly with time and were controlled by a non-SHA calcium phosphate phase(s) with low Ca/P solution molar ratios (0.91-1.51) relative to solid SHA ratios (1.56-1.64). At later times, Ca/P solution molar ratios (1.47-1.79) were near solid SHA ratios and release rates decreased slowly indicating that SHA controlled Ca and P release. Substituted SHA materials had faster dissolution rates relative to unsubstituted SHA. The initial metal release rate order was Mn >> Cu > Fe which followed metal-oxide/phosphate solubility suggesting that poorly crystalline metal-oxides/phosphates were dominating metal release. Similar metal release rates for all substituted SHA (approximately 0.01 cmol kg-1 min-1) at the end of the DTPA experiment indicated that SHA dissolution was supplying the metals into solution and that poorly crystalline metal-oxide/phosphates were not controlling metal release. Results indicate that non-SHA Ca-phosphate phases and poorly crystalline metal-oxide/phosphates will contribute Ca, P, and metals. After these phases have dissolved, substituted SHA will be the source of Ca, P, and metals for plants.

Countermeasure for space flight effects on immune system: nutritional nucleotides.

Microgravity and its environment have adverse effects on the immune system. Abnormal immune responses observed in microgravity may pose serious consequences, especially for the recent directions of NASA for long-term space missions to Moon, Mars and deep Space exploration. The study of space flight immunology is limited due to relative inaccessibility, difficulty of performing experiments in space, and inadequate provisions in this area in the United States and Russian space programs (Taylor 1993). Microgravity and stress experienced during space flights results in immune system aberration (Taylor 1993). In ground-based mouse models for some of the microgravity effects on the human body, hindlimb unloading (HU) has been reported to cause abnormal cell proliferation and cytokine production (Armstrong et al., 1993, Chapes et al. 1993). In this report, we document that a nutritional nucleotide supplementation as studied in ground-based microgravity analogs, has potential to serve as a countermeasure for the immune dysfunction observed in space travel.

Cardiac health for astronauts: current selection standards and their limitations.

INTRODUCTION: The screening tests for coronary artery disease (CAD) for applicants and the active astronaut corps are similar to those performed in the 1960s. Due to the limited treatment and return capabilities of most space vehicles, an in-flight cardiac event would result in mission failure. Improved CAD screening of astronauts is, therefore, paramount to long-duration mission success. METHODS: Literature review was performed to compare active and retired astronaut populations to other asymptomatic low-risk cohorts. All populations were examined to determine the incidence and prevalence of CAD. Framingham risk scores were calculated in NASA's active and retired astronaut corps and compared with age- and gender-matched controls. RESULTS: The current standards used for astronaut selection have been successful in creating a cohort that has less risk than their age- and gender-matched counterparts from the general population. However, the existing astronaut cardiovascular screening and selection tests do not adequately rule out CAD for long-duration missions, and, therefore, a "significant" risk of cardiac event remains, especially as we look toward Exploration Class missions. CONCLUSIONS: The current astronaut selection and retention standards may not adequately prevent cardiac events from occurring with the longer duration flights. Future research should be directed toward increasing the primary and secondary prevention of CAD in the astronaut cohort. In the meantime, the space program should evaluate the use of more aggressive terrestrial screening tools. It is important not to remove all older, experienced pilots from spaceflight crews unless overt or predictable pathology has been clearly identified.

A complete dielectric response model for liquid water: a solution of the Bethe ridge problem.

We present a complete yet computationally simple model for the dielectric response function of liquid water over the energy-momentum plane, which, in contrast to earlier models, is consistent with the recent inelastic X-ray scattering spectroscopy data at both zero and finite momentum transfer values. The model follows Ritchie's extended-Drude algorithm and is particularly effective at the region of the Bethe ridge, substantially improving previous models. The present development allows for a more accurate simulation of the inelastic scattering and energy deposition process of low-energy electrons in liquid water and other biomaterials. As an example, we calculate the stopping power of liquid water for electrons over the 0.1-10 keV range where direct experimental measurements are still impractical and the Bethe stopping formula is inaccurate. The new stopping power values are up to 30-40% lower than previous calculations. Within the range of validity of the first Born approximation, the new values are accurate to within the experimental uncertainties (a few percent). At the low end, the introduction of Born corrections raises the uncertainty to perhaps approximately 10%. Thus the present model helps extend the ICRU electron stopping power database for liquid water down to about two orders of magnitude with a comparable level of uncertainty.

Reduced gravity evaluation of potential spaceflight-compatible flow cytometer technology.

BACKGROUND: The presence of a spaceflight-compatible flow cytometer onboard the International Space Station would be extremely beneficial for in-flight medicine and reduced gravity research. In the past, commercially available flow cytometers were not suitable for use during spaceflight due to their size, weight, power, and sheath fluid requirements. Recent advances in cytometer technology have enabled features that now make a spaceflight-compatible cytometer possible. We evaluated a small, robust novel cytometer with design features that minimize or eliminate many incompatibilities with spaceflight. This cytometer is highly miniaturized and lightweight, does not require sheath fluid, and uses a low-energy diode laser. The ability to achieve laminar particle flow without sheath fluid is important because an instrument using large liquid volumes and producing an equivalent amount of hazardous liquid waste would not meet spaceflight constraints. METHODS: For this study the cytometer was modified so that stained liquid cell samples could be delivered during reduced gravity. The cytometer was then evaluated onboard the NASA KC-135 reduced gravity research aircraft. The KC-135 uses a parabolic flight path to generate essentially zero gravity conditions for 30-s increments. Typically 40 parabolas are flown per mission, resulting in approximately 20 min of reduced gravity during which research may be performed. During this evaluation, bead-based cytometer precision, photomultiplier tube linearity, and leukocyte immunophenotype analysis were performed during reduced gravity. The flight data were then compared with ground-based control data and data generated using a reference cytometer (Beckman-Coulter XL). RESULTS: This novel cytometer functioned well during reduced gravity and produced data comparable to those of ground-based controls with only minor caveats. The reduced gravity cell immunophenotype data were indistinguishable from ground control data and reference cytometric data. Bead-based instrument precision (coefficient of variation) was slightly increased during reduced gravity operation, but not to a degree that would affect most common flow cytometric applications. The ability of the instrument to collect absolute cell counts was validated. CONCLUSIONS: This study represents the first generation of real-time flow cytometry data during zero gravity. With modifications, the evaluated cytometer technology could be the basis from which an operational spaceflight-compatible flow cytometer is developed.

A procedure for benchmarking laboratory exposures with 1 A GeV iron ions.

A new version of the HZETRN code capable of simulating HZE ions with either laboratory or space boundary conditions is under development. The computational model consists of combinations of physical perturbation expansions based on the scales of atomic interaction, multiple scattering, and nuclear reactive processes with use of asymptotic/Neumann expansions with non-perturbative corrections. The code contains energy loss with straggling, nuclear attenuation, nuclear fragmentation with energy dispersion and downshifts, and off-axis dispersion with multiple scattering under preparation. The present benchmark is for a broad directed beam for 1 A GeV iron ion beams with 2 A MeV width and four targets of polyethylene, polymethyl metachrylate, aluminum, and lead of varying thickness from 5 to 30 g/cm2. The benchmark quantities will be dose, track averaged LET, dose averaged LET, fraction of iron ion remaining, and fragment energy spectra after 23 g/cm2 of polymethyl metachrylate.

Computational methods for the HZETRN code.

Asymptotic expansion has been used to simplify the transport of high charge and energy ions for broad beam applications in the laboratory and space. The solution of the lowest order asymptotic term is then related to a Green's function for energy loss and straggling coupled to nuclear attenuation providing the lowest order term in a rapidly converging Neumann series for which higher order collisions terms are related to the fragmentation events including energy dispersion and downshift. The first and second Neumann corrections were evaluated numerically as a standard for further analytic approximation. The first Neumann correction is accurately evaluated over the saddle point whose width is determined by the energy dispersion and located at the downshifted ion collision energy. Introduction of the first Neumann correction leads to significant simplification of the second correction term allowing application of the mean value theorem and a second saddle point approximation. The regular dependence of the second correction spectral dependence lends hope to simple approximation to higher corrections. At sufficiently high energy nuclear cross-section variations are small allowing non-perturbative methods to all orders and renormalization of the second corrections allow accurate evaluation of the full Neumann series.

Validation of the HZETRN code for laboratory exposures with 1A GeV iron ions in several targets.

A new version of the HZETRN code capable of validation with HZE ions in either the laboratory or the space environment is under development. The computational model consists of the lowest order asymptotic approximation followed by a Neumann series expansion with non-perturbative corrections. The physical description includes energy loss with straggling, nuclear attenuation, nuclear fragmentation with energy dispersion and downshift. Measurements to test the model were performed at the Alternating Gradient Synchrotron and the NASA Space Radiation Laboratory at Brookhaven National Laboratory with iron ions. Surviving beam particles and produced fragments were measured with solid-state detectors. Beam analysis software has been written to relate the computational results to the measured energy loss spectra of the incident ions for rapid validation of modeled target transmission functions.

High LET-induced H2AX phosphorylation around the Bragg curve.

We investigated the spatial distribution of the induction of the phosphorylated form of the histone protein H2AX (gamma-H2AX), known to be activated by DSBs. Following irradiation of human fibroblast cells with 600 MeV/nucleon silicon and 600 MeV/nucleon iron ions we observed the formation of gamma-H2AX aggregates in the shape of streaks stretching over several micrometers in an x/y plane. Polyethylene shielding was used to achieve a Bragg curve distribution with beam geometry parallel to the monolayer of cells. We present data that highlights the formation of immunofluorescent gamma-H2AX tracks showing the ion trajectories across the Bragg peak of irradiated human fibroblast cells. Qualitative analyses of these distributions indicated potentially increased clustering of DNA damage before the Bragg peak, enhanced gamma-H2AX distribution at the peak, and provided visual evidence of high-linear energy transfer particle traversal of cells beyond the Bragg peak in agreement with one-dimensional transport approximations. Spatial assessment of gamma-H2AX fluorescence may provide direct insights into DNA damage across the Bragg curve for high charge and energy ions including the biological consequences of shielding and possible contributors to bystander effects.

Comparison of genotoxic damage in monolayer cell cultures and three-dimensional tissue-like cell assemblies.

Assessing the biological risks associated with exposure to the high-energy charged particles encountered in space is essential for the success of long-term space exploration. Although prokaryotic and eukaryotic cell models developed in our laboratory and others have advanced our understanding of many aspects of genotoxicity, in vitro models are needed to assess the risk to humans from space radiation insults. Such models must be representative of the cellular interactions present in tissues and capable of quantifying genotoxic damage. Toward this overall goal, the objectives of this study were to examine the effect of the localized microenvironment of cells, cultured as either 2-dimensional (2D) monolayers or 3-dimensional (3D) aggregates, on the rate and type of genotoxic damage resulting from exposure to Fe-charged particles, a significant portion of space radiation. We used rodent transgenic cell lines containing 50-70 copies of a LacI transgene to provide the enhanced sensitivity required to quantify mutational frequency and type in the 1100-bp LacI target as well as assessment of DNA damage to the entire 45-kbp construct. Cultured cells were exposed to high energy Fe charged particles at Brookhaven National Laboratory's Alternating Gradient Synchrotron facility for a total dose ranging from 0.1 to 2 Gy and allowed to recover for 0-7 days, after which mutational type and frequency were evaluated. The mutational frequency was found to be higher in 3D samples than in 2D samples at all radiation doses. Mutational frequency also was higher at 7 days after irradiation than immediately after exposure. DNA sequencing of the mutant targets revealed that deletional mutations contributed an increasingly high percentage (up to 27%) of all mutations in cells as the dose was increased from 0.5 to 2 Gy. Several mutants also showed large and complex deletions in multiple locations within the LacI target. However, no differences in mutational type were found between the 2D and the 3D samples. These 3D tissue-like model systems can reduce the uncertainty involved in extrapolating risk between in vitro cellular and in vivo models.

Vestibular and non-vestibular contributions to eye movements that compensate for head rotations during viewing of near targets.

Geometry dictates that when subjects view a near target during head rotation the eyes must rotate more than the head. The relative contribution to this compensatory response by adjustment of the vestibulo-ocular reflex gain (Gvor), visual tracking mechanisms including prediction, and convergence is debated. We studied horizontal eye movements induced by sinusoidal 0.2-2.8 Hz, en-bloc yaw rotation as ten normal humans viewed a near target that was either earth-fixed (EFT) or head-fixed (HFT). For EFT, group median gain was 1.49 at 0.2 Hz declining to 1.08 at 2.8 Hz. For HFT, group median gain was 0.03 at 0.2 Hz increasing to 0.71 at 2.8 Hz. By applying transient head perturbations (peak acceleration >1,000 degrees s(-2)) during sinusoidal rotation, we determined that Gvor was similar during either EFT or HFT conditions, and contributed only approximately 75% to the compensatory response. We confirmed that retinal image slip contributed to the compensatory response by demonstrating reduced gain during EFT viewing under strobe illumination. Gain also declined during sum-of-sines head rotations, confirming the contribution of predictive mechanisms. The gain of compensatory eye movements was similar during monocular or binocular viewing, although vergence angle was greater during binocular viewing. Comparison with previous studies indicates that mechanisms for generation of eye rotations during near viewing depend on head stimulus type (rotation or translation), waveform (transient or sinusoidal), and the species being tested.