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Surface characteristics of spacecraft components affect the aggregation of microorganisms and may lead to different survival rates of bacteria on Mars landers.

Layers of dormant endospores of Bacillus subtilis HA101 were applied to eight different spacecraft materials and exposed to martian conditions of low pressure (8.5 mbar), low temperature (-10 degrees C), and high CO(2) gas composition and irradiated with a Mars-normal ultraviolet (UV-visible- near-infrared spectrum. Bacterial layers were exposed to either 1 min or 1 h of Mars-normal UV irradiation, which simulated clear-sky conditions on equatorial Mars (0.1 tau). When exposed to 1 min of Mars UV irradiation, the numbers of viable endospores of B. subtilis were reduced three to four orders of magnitude for two brands of aluminum (Al), stainless steel, chemfilm-treated Al, clear-anodized Al, and black-anodized Al coupons. In contrast, bacterial survival was reduced only one to two orders of magnitude for endospores on the non-metal materials astroquartz and graphite composite when bacterial endospores were exposed to 1 min of Mars UV irradiation. When bacterial monolayers were exposed to 1 h of Mars UV irradiation, no viable bacteria were recovered from the six metal coupons listed above. In contrast, bacterial survival was reduced only two to three orders of magnitude for spore layers on astroquartz and graphite composite exposed to 1 h of Mars UV irradiation. Scanning electron microscopy images of the bacterial monolayers on all eight spacecraft materials revealed that endospores of B. subtilis formed large aggregates of multilayered spores on astroquartz and graphite composite, but not on the other six spacecraft materials. It is likely that the formation of multilayered aggregates of endospores on astroquartz and graphite composite is responsible for the enhanced survival of bacterial cells on these materials.

Bacillus subtilis↗

Acceleration in other axes affects +Gz tolerance: dynamic centrifuge simulation of agile flight.

BACKGROUND: Modern, thrust-vectored jet aircraft have the capability of developing multi-axis accelerations, especially during the performance of "supermaneuvers." These "agile" aircraft are capable of unconventional flight. The human consequences of this agile flight environment are unknown. METHODS: This multi-axis acceleration environment was studied on the Dynamic Environment Simulator gimbaled centrifuge. There were nine relaxed, unprotected subjects who were exposed to either lateral (+/- 1, +/- 2 Gy), transverse chest-to-back (+1, 2.5, or 4 Gx), or back-to-chest (-1 Gx) sustained acceleration. Positive C (+Gz) acceleration was then added beginning at 1.0 Gz by gradual onset (0.1 Gz x s(-1)) until the subjects lost nearly all of their vision. Baseline +Gz-only relaxed tolerances were measured before and after all combined Gy/Gz and Gx/Gz exposures. Heart rate, percent cerebral oxygen saturation, and cerebral blood volumes were collected during each exposure. RESULTS: Adding moderate transverse (+Gx) acceleration significantly reduced +Gz tolerance. Relaxed, unprotected +Gz tolerance was reduced approximately 0.25 G when 1.0 or 2.5 Gx was added to the increasing +Gz exposure. Adding moderate lateral Gy significantly increased +Gz tolerance. Relaxed, unprotected +Gz tolerance was increased approximately 0.5 G when +2 Gy or -2 Gy was added to the -Gz exposure. The decrease in cerebral blood volume was significantly less when +Gz was added to -1 Gx compared with the addition of +Gz to +Gx. CONCLUSIONS: Multi-axis sustained accelerations, such as those experienced during thrust-vectored aircraft maneuvers, can either enhance or reduce the +Gz tolerance of the pilot depending on the direction of the net gravitoinertial force. Gy acceleration in conjunction with Gz acceleration can enhance G tolerance. Gx acceleration in addition to Gz acceleration can reduce G tolerance.

Acceleration↗

Work capability during isolation.

The aim of this study was to investigate the effects of prolonged isolation on the higher psychic functions, like working memory, attention concentration, and intellect (problem solving and decision making), and on sensory-motor skills and stress resistance. Previous Soviet simulation studies and the ISEMSI isolation experiment have indicated that prolonged isolation can affect higher psychic functions. A set of psychological tests in the form of a computer game was presented each workday to the chamber crew and to the ground crew serving as a control group. In analyzing the data it was taken into account that performance can be affected not only by the influence of isolation, but also by a learning process and by subject motivation. In addition, a distinction was made between absolute score and stability (range) of the score. Analysis of the chamber crew's work capability as a function of time showed the occurrence of three distinct periods of adaptation: (1) a period of acute adaptation in week 1, (2) a period of stable adaptation during weeks 3-6, and (3) a period of "final effort" in weeks 8-9. While in general the effect of isolation on the absolute scores was minor, larger ranges for the scores in "working memory," "attention concentration," and "calculation under time deficit" tests are an indication of increased instability, probably due to stress resistance. The 4 female subjects of the combined groups scored significantly higher than the 5 males in "attention concentration/distribution," "spatial orientation," "intuition in visual search," and "logical decision making under time deficit." Males presented higher scores in "calculation under time deficit" and working memory, and higher stability in "attention concentration" and "calculation under time deficit."

Adaptation, Psychological↗

Dietary salt and urinary calcium excretion in a human bed rest spaceflight model.

BACKGROUND: Dietary salt is known to increase the excretion of urinary calcium (Ca). To determine the potential role of dietary sodium (Na) on the calciuria associated with a spaceflight simulation model, we evaluated urinary Ca in two groups of bed rest subjects fed either high or low normal amounts of salt. METHODS: We analyzed urinary Ca excretion expressed in terms of creatinine (UCa/Cr), fractional Ca excretion (FECa), and urinary cAMP (UCAMP) as an index of parathyroid function, in the urine of 30-50-yr-old male volunteers for 6 degrees head down tilt bed rest studies. Dietary Na was in the high normal range (190 mmol x d(-1)) in 8 men for 7 d (HiNa), and in the low normal range (114 mmol x d(-1)) in 11 men for 30 d (LoNa) bed rest. Dietary Ca averaged 20 mmol x d(-1) in both studies. RESULTS: Within the first 3 bed rest days, subjects in the HiNa study showed increases in UCa/Cr (0.1130 +/- 0.05 to 0.161 +/- 0.05, p < 0.002) and in FECa (1.95 +/- 0.70 to 3.19 +/- 0.93, p < 0.001); those in LoNa showed no change in UCa/Cr (0.125 +/- 0.06 to 0.121 +/- 0.07, NS) or FECa (1.93 +/- 0.75 to 2.22 +/- 0.63). After the 5th bed rest day UCa/Cr stabilized at similar levels in both dietary groups. UCAMP decreased 20% during the first week of bed rest with HiNa, but not until the third week with LoNa diets (p < 0.05). CONCLUSION: These findings implicate high salt diets in Ca excretion in a spaceflight model and suggest that low normal salt diets may reduce early calciuria associated with spaceflight.

Adult↗

Skeletal muscle satellite cells cultured in simulated microgravity.

Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and, therefore, provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75% satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D culture and 3-D HARV culture. Plating efficiency (cells attached divided by cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and nonsatellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability because glucose levels in medium from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARV were joined together by cells into 3-D aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of satellite cells on microcarrier beads within the HARV bioreactor results in a 3-D level of organization that could provide a more suitable model to study postnatal muscle development than is currently available with standard culture methods.

Animals↗

Neonatal rat heart cells cultured in simulated microgravity.

In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two-dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by nonmyocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA-designed High-Aspect Ratio Vessel (HARV) bioreactors provide a low shear environment that allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells cultured in HARVs adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARVs using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar; however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissuelike organization of cardiac cells in vitro.

Animals↗

Survival of endospores of Bacillus subtilis on spacecraft surfaces under simulated martian environments: implications for the forward contamination of Mars.

Experiments were conducted in a Mars simulation chamber (MSC) to characterize the survival of endospores of Bacillus subtilis under high UV irradiation and simulated martian conditions. The MSC was used to create Mars surface environments in which pressure (8.5 mb), temperature (-80, -40, -10, or +23 degrees C), gas composition (Earth-normal N2/O2 mix, pure N2, pure CO2, or a Mars gas mix), and UV-VIS-NIR fluence rates (200-1200 nm) were maintained within tight limits. The Mars gas mix was composed of CO2 (95.3%), N2 (2.7%), Ar (1.7%), O2 (0.2%), and water vapor (0.03%). Experiments were conducted to measure the effects of pressure, gas composition, and temperature alone or in combination with Mars-normal UV-VIS-NIR light environments. Endospores of B. subtilis, were deposited on aluminum coupons as monolayers in which the average density applied to coupons was 2.47 x 10(6) bacteria per sample. Populations of B. subtilis placed on aluminum coupons and subjected to an Earth-normal temperature (23 degrees C), pressure (1013 mb), and gas mix (normal N2/O2 ratio) but illuminated with a Mars-normal UV-VIS-NIR spectrum were reduced by over 99.9% after 30 sec exposure to Mars-normal UV fluence rates. However, it required at least 15 min of Mars-normal UV exposure to reduce bacterial populations on aluminum coupons to non-recoverable levels. These results were duplicated when bacteria were exposed to Mars-normal environments of temperature (-10 degrees C), pressure (8.5 mb), gas composition (pure CO2), and UV fluence rates. In other experiments, results indicated that the gas composition of the atmosphere and the temperature of the bacterial monolayers at the time of Mars UV exposure had no effects on the survival of bacterial endospores. But Mars-normal pressures (8.5 mb) were found to reduce survival by approximately 20-35% compared to Earth-normal pressures (1013 mb). The primary implications of these results are (a) that greater than 99.9% of bacterial populations on sun-exposed surfaces of spacecraft are likely to be inactivated within a few tens of seconds to a few minutes on the surface of Mars, and (b) that within a single Mars day under clear-sky conditions bacterial populations on sun-exposed surfaces of spacecraft will be sterilized. Furthermore, these results suggest that the high UV fluence rates on the martian surface can be an important resource in minimizing the forward contamination of Mars.

Atmospheric Pressure↗

A model for the mechanism of optic nerve sheath fenestration.

The mechanism by which optic nerve sheath fenestration relieves papilledema associated with increased intracranial pressure has not been clearly defined. A model was constructed to determine if the induction of fluid flow along the nerve sheath and through the fenestration could account for a reduction in pressure around the optic nerve, as might be expected according to Bernoulli's equation of fluid dynamics. The model states that as the velocity of a fluid increases, the pressure it exerts decreases. The model simulated fluid spaces corresponding to the intracranial vault, chiasm, and optic nerves. The unfenestrated model showed direct transmission of elevated intracranial pressure to the nerves, consistent with the production of papilledema. When one nerve was fenestrated, fluid flow along the nerve was initiated and pressure in that nerve sheath dropped substantially. In addition, pressure in the unfenestrated sheath dropped due to fluid communication across the chiasm. These lower intrasheath pressures were consistent with the bilateral resolution of papilledema after unilateral fenestration. The reduced intrasheath pressures persisted even when the intracranial pressure was again elevated to pathologic levels as long as active fluid flow continued along the nerve sheaths. Thus, induction of cerebrospinal fluid flow along the optic nerve sheath by fenestration appears to locally reduce the pressure around the optic nerve, in spite of persistently elevated intracranial pressure. This is in accordance with what would be predicted by Bernoulli's equation.

Cerebrospinal Fluid↗

Gibbs adsorption isotherm combined with Monte Carlo sampling to see action of cosolutes on protein folding.

Driven by conditions set by smaller solutes, proteins fold and unfold. Experimentally, these conditions are stated as intensive variables--pH and other chemical potentials--as though small solutes were infinite resources that come at an externally varied free energy cost. Computationally, the finite spaces of simulation allow only fixed numbers of these solutes. By combining the analytic Gibbs adsorption isotherm with the computational Monte Carlo sampling of polymer configurations, we have been able to overcome an inherent limitation of computer simulation. The idea is to compute analytically the free energy changes wrought by solutes on each particular configuration. Then numerical computation is needed only to sample the set of configurations as efficiently as when no bathing solute is present. For illustration, the procedure is applied to an idealized two-dimensional heteropolymer to yield lessons about the effect of cosolutes on protein stability.

Adsorption↗

Fast and simple Monte Carlo algorithm for side chain optimization in proteins: application to model building by homology.

An unknown protein structure can be predicted with fair accuracy once an evolutionary connection at the sequence level has been made to a protein of known 3-D structure. In model building by homology, one typically starts with a backbone framework, rebuilds new loop regions, and replaces nonconserved side chains. Here, we use an extremely efficient Monte Carlo algorithm in rotamer space with simulated annealing and simple potential energy functions to optimize the packing of side chains on given backbone models. Optimized models are generated within minutes on a workstation, with reasonable accuracy (average of 81% side chain chi 1 dihedral angles correct in the cores of proteins determined at better than 2.5 A resolution). As expected, the quality of the models decreases with decreasing accuracy of backbone coordinates. If the back-bone was taken from a homologous rather than the same protein, about 70% side chain chi 1 angles were modeled correctly in the core in a case of strong homology and about 60% in a case of medium homology. The algorithm can be used in automated, fast, and reproducible model building by homology.

Algorithms↗

Gravitational loading of a simulated launch alters mRNA expression in osteoblasts.

Serum-deprived mouse osteoblastic cells (MC3T3-E1a) were centrifuged under a regime designed to simulate a space shuttle launch (maximum of 3g). Messenger RNA levels for eight genes involved in bone growth and maintenance were determined using RT-PCR. Following 30 min of centrifugation, mRNA level for early response gene c-fos was significantly increased 89% (P < 0.05). The c-fos induction was transient and returned to control levels after 3 h. The mRNA level for the mineralization marker gene osteocalcin was significantly decreased to 44% of control level (P < 0.005) 3 h after centrifugation. No changes in mRNA levels were detected for c-myc, TGFbeta1, TGFbeta2, cyclophilin A, or actin. No basal mRNA level for TGFbeta3 was detected. In addition, no change in the steady-state synthesis of prostaglandin E2 was detected, possibly due to lack of lipid substrates in serum-deprived cells, suggesting that the increase in c-fos mRNA in response to gravitational loading is a result of mechanical stimulation. These results indicate that a small magnitude mechanical loading, such as that experienced during a shuttle launch, can alter mRNA levels in quiescent osteoblastic cells.

Actins↗

The effect of space flight on monoclonal antibody synthesis in a hybridoma mouse cell line.

The hybridoma cell line, 3G10G5, producing a monoclonal antibody to the major capsid protein VP1 from the avian polyomavirus budgerigar fledgling disease virus, was produced from a Balb/C mouse. This cell line was used to test the effects of microgravity on cellular processes, specifically protein synthesis. A time course study utilizing incorporation of [35S]methionine into newly synthesized monoclonal antibody was performed on STS-77. After 5.5 days, it was observed that cell counts for the samples exposed to microgravity were lower than those of ground-based samples. However, radiolabel incorporation of the synthesized monoclonal antibody was similar in both orbiter and ground control samples. Overall, microgravity does not seem to have an effect on this cell line's ability to synthesize IgG protein.

Animals↗

Competing species model with behavioral adaptation.

This paper studies the properties of a modified Lotka-Volterra model for two competing species, in which the coefficients of the interaction terms are time-dependent averages of the level of interaction over the entire past. For this model, it is shown that competitive exclusion does not occur, there are two possible stable equilibrium points, and in a certain region of parameter space numerical simulations suggest the existence of interesting oscillatory solutions.

Adaptation, Psychological↗

Analysis of human torso motion with muscle actuators.

A biomechanical model is devised to analyze the motion of the human torso and estimate the load on the lumbar spine, the contraction forces in the trunk muscles, and the sensory signals between the muscle and the nervous system. A state space formulation of three-dimensional (3D) equations of motion of the human torso is presented with muscle forces as input to the system. At least 3 pairs of skeletal muscles are considered to be necessary for 3D motion of human torso. Functional anatomy of these major muscles of the human trunk are discussed. These muscles as well as their feedback and feedforward sensory paths are modeled by linear viscoelastic components and force generators. Stability of the torso with three pairs of muscles is studied both with muscle spindles inactive (open loop) and active (closed loop). Voluntary point-to-point motion of the torso in three-dimensional space is simulated on a digital computer, employing a dynamic controller, where feedback gains are tuned (programmed) by higher centers of nervous system.

Biomechanical Phenomena↗

Modeling locomotor dysfunction following spaceflight with Galvanic vestibular stimulation.

In this study locomotor and gaze dysfunction commonly observed in astronauts following spaceflight were modeled using two Galvanic vestibular stimulation (GVS) paradigms: (1) pseudorandom, and (2) head-coupled (proportional to the summed vertical linear acceleration and yaw angular velocity obtained from a head-mounted Inertial Measurement Unit). Locomotor and gaze function during GVS were assessed by tests previously used to evaluate post-flight astronaut performance; dynamic visual acuity (DVA) during treadmill locomotion at 80 m/min, and navigation of an obstacle course. During treadmill locomotion with pseudorandom GVS there was a 12% decrease in coherence between head pitch and vertical translation at the step frequency relative to the no GVS condition, which was not significantly different to the 15% decrease in coherence observed in astronauts following shuttle missions. This disruption in head stabilization likely resulted in a decrease in DVA equivalent to the reduction in acuity observed in astronauts 6 days after return from extended missions aboard the International Space Station (ISS). There were significant increases in time-to-completion of the obstacle course during both pseudorandom (21%) and head-coupled (14%) GVS, equivalent to an ISS astronaut 5 days post-landing. An attempt to suppress head movement was evident during both pseudorandom and head-coupled GVS while negotiating the obstacle course, with a 20 and 16%, decrease in head pitch and yaw velocity, respectively. The results of this study demonstrate that pseudorandom GVS generates many of the salient features of post-flight locomotor dysfunction observed in astronauts following short and long duration missions. An ambulatory GVS system may prove a useful adjunct to the current pre-flight astronaut training regimen.

Adult↗

A simple model illustrating the role of turbulence on phytoplankton blooms.

The problem of the vertical distribution of phytoplankton is considered in the presence of gravitational settling, turbulent mixing, population growth due to cell division and a constant rate of loss due to predation and natural death. Nutrients are assumed to be plentiful so that the production rate depends only on the light available for photosynthesis. The non-linear saturation of plankton growth is modeled by allowing the attenuation rate of light to be a linear function of the plankton density. The turbulent diffusivity is assumed constant which corresponds to a mixed layer depth very much greater than the depth of light penetration (euphotic depth). It is shown that an exact analytical solution of this non-linear problem is possible for an idealized model in which the functional dependence of production on light intensity is assumed to be a step function. Non-zero solutions are shown to exist only if the parameters characterizing the system are above a certain critical curve in a two dimensional parameter space. Numerical simulations using functional forms of the production curve that resemble the measured photosynthetic response of plankton, show, that the qualitative behavior of the system is similar to that of the idealized model presented. Comparisons are made with other analytical approaches to the problem.

Computer Simulation↗

In vitro cleaning efficacy and resistance to insertion test of interdental brushes.

The aim of this study was to compare the resistance to insertion forces and cleaning efficacy values of triangular interdental brushes (TIB) and conventional round interdental brushes (RIB). Extracted human teeth were fixed in a socket and matched to pairs simulating interdental spaces. Before and after standardized cleaning of the proximal surfaces, the pre- and postbrushing situations were registered with a digital camera. The cleaning efficacy was quantified by digital image subtraction. The forces necessary for insertion were measured with a load cell. The ratio between the relative cleaning efficacy and the forces necessary for insertion was calculated. The results of our study revealed that the relative cleaning efficacies of round and TIB were equal. There were no statistically significant differences. TIB showed significantly lower resistance to insertion values. Consequently, at identical resistance to insertion the TIB cleaned more effectively compared with the RIB, which means that the same effectiveness was reached by significantly lower forces at insertion. TIB showed a more favorable ratio between relative cleaning efficacy and resistance to insertion than RIB.

Dental Devices, Home Care↗

On the passive cardiac conductivity.

In order to relate the structure of cardiac tissue to its passive electrical conductivity, we created a geometrical model of cardiac tissue on a cellular scale that encompassed myocytes, capillaries, and the interstitial space that surrounds them. A special mesh generator was developed for this model to create realistically shaped myocytes and interstitial space with a controlled degree of variation included in each model. In order to derive the effective conductivities, we used a finite element model to compute the currents flowing through the intracellular and extracellular space due to an externally applied electrical field. The product of these computations were the effective conductivity tensors for the intracellular and extracellular spaces. The simulations of bi-domain conductivities for healthy tissue resulted in an effective intracellular conductivity of 0.16S/m (longitudinal) and 0.005 S/m (transverse) and an effective extracellular conductivity of 0.21S/m (longitudinal) and 0.06 S/m (transverse). The latter values are within the range of measured values reported in literature. Furthermore, we anticipate that this method can be used to simulate pathological conditions for which measured data is far more sparse.

Electric Conductivity↗