Showing papers in "Life sciences in space research in 2014"

Space radiation risks to the central nervous system

Abstract: Central nervous system (CNS) risks which include during space missions and lifetime risks due to space radiation exposure are of concern for long-term exploration missions to Mars or other destinations. Possible CNS risks during a mission are altered cognitive function, including detriments in short-term memory, reduced motor function, and behavioral changes, which may affect performance and human health. The late CNS risks are possible neurological disorders such as premature aging, and Alzheimer's disease (AD) or other dementia. Radiation safety requirements are intended to prevent all clinically significant acute risks. However the definition of clinically significant CNS risks and their dependences on dose, dose-rate and radiation quality is poorly understood at this time. For late CNS effects such as increased risk of AD, the occurrence of the disease is fatal with mean time from diagnosis of early stage AD to death about 8 years. Therefore if AD risk or other late CNS risks from space radiation occur at mission relevant doses, they would naturally be included in the overall acceptable risk of exposure induced death (REID) probability for space missions. Important progress has been made in understanding CNS risks due to space radiation exposure, however in general the doses used in experimental studies have been much higher than the annual galactic cosmic ray (GCR) dose (∼0.1 Gy/y at solar maximum and ∼0.2 Gy/y at solar minimum with less than 50% from HZE particles). In this report we summarize recent space radiobiology studies of CNS effects from particle accelerators simulating space radiation using experimental models, and make a critical assessment of their relevance relative to doses and dose-rates to be incurred on a Mars mission. Prospects for understanding dose, dose-rate and radiation quality dependencies of CNS effects and extrapolation to human risk assessments are described.

Biological effects of space radiation and development of effective countermeasures

Abstract: As part of a program to assess the adverse biological effects expected from astronaut exposure to space radiation, numerous different biological effects relating to astronaut health have been evaluated. There has been major focus recently on the assessment of risks related to exposure to solar particle event (SPE) radiation. The effects related to various types of space radiation exposure that have been evaluated are: gene expression changes (primarily associated with programmed cell death and extracellular matrix (ECM) remodeling), oxidative stress, gastrointestinal tract bacterial translocation and immune system activation, peripheral hematopoietic cell counts, emesis, blood coagulation, skin, behavior/fatigue (including social exploration, submaximal exercise treadmill and spontaneous locomotor activity), heart functions, alterations in biological endpoints related to astronaut vision problems (lumbar puncture/intracranial pressure, ocular ultrasound and histopathology studies), and survival, as well as long-term effects such as cancer and cataract development. A number of different countermeasures have been identified that can potentially mitigate or prevent the adverse biological effects resulting from exposure to space radiation.

Space radiation protection: Destination Mars

Abstract: National space agencies are planning a human mission to Mars in the XXI century. Space radiation is generally acknowledged as a potential showstopper for this mission for two reasons: a) high uncertainty on the risk of radiation-induced morbidity, and b) lack of simple countermeasures to reduce the exposure. The need for radiation exposure mitigation tools in a mission to Mars is supported by the recent measurements of the radiation field on the Mars Science Laboratory. Shielding is the simplest physical countermeasure, but the current materials provide poor reduction of the dose deposited by high-energy cosmic rays. Accelerator-based tests of new materials can be used to assess additional protection in the spacecraft. Active shielding is very promising, but as yet not applicable in practical cases. Several studies are developing technologies based on superconducting magnetic fields in space. Reducing the transit time to Mars is arguably the best solution but novel nuclear thermal-electric propulsion systems also seem to be far from practical realization. It is likely that the first mission to Mars will employ a combination of these options to reduce radiation exposure.

Significant reduction in energy for plant-growth lighting in space using targeted LED lighting and spectral manipulation

Abstract: Bioregenerative life-support systems involving photoautotrophic organisms will be necessary to sustain long-duration crewed missions at distant space destinations. Since sufficient sunlight will not always be available for plant growth at many space destinations, efficient electric-lighting solutions are greatly needed. The present study demonstrated that targeted plant lighting with light-emitting diodes (LEDs) and optimizing spectral parameters for close-canopy overhead LED lighting allowed the model crop leaf lettuce ( Lactuca sativa L. cv. ‘Waldmann's Green’) to be grown using significantly less electrical energy than using traditional electric-lighting sources. Lettuce stands were grown hydroponically in a growth chamber controlling temperature, relative humidity, and CO 2 level. Several red:blue ratios were tested for growth rate during the lag phase of lettuce growth. In addition, start of the exponential growth phase was evaluated. Following establishment of a 95% red + 5% blue spectral balance giving the best growth response, the energy efficiency of a targeted lighting system was compared with that of two total coverage (untargeted) LED lighting systems throughout a crop-production cycle, one using the same proportion of red and blue LEDs and the other using white LEDs. At the end of each cropping cycle, whole-plant fresh and dry mass and leaf area were measured and correlated with the amount of electrical energy (kWh) consumed for crop lighting. Lettuce crops grown with targeted red + blue LED lighting used 50% less energy per unit dry biomass accumulated, and the total coverage white LEDs used 32% less energy per unit dry biomass accumulated than did the total coverage red + blue LEDs. An energy-conversion efficiency of less than 1 kWh/g dry biomass is possible using targeted close-canopy LED lighting with spectral optimization. This project was supported by NASA grant NNX09AL99G.

Advances in NASA radiation transport research: 3DHZETRN

Abstract: The computationally efficient HZETRN code has been used in recent trade studies for lunar and Martian exploration and is currently being used in the engineering development of the next generation of space vehicles, habitats, and extra vehicular activity equipment. Code development has been based on a progression of approximations first assuming all particles are produced in the initiator direction of incidence (straight-ahead) later improved by treating neutrons produced in the backward hemisphere as moving straight-back (bi-directional). A new version (3DHZETRN) capable of transporting High charge (Z) and Energy (HZE) and light ions (including neutrons) under space-like boundary conditions with enhanced neutron and light ion propagation in transverse directions is developed. Herein, new algorithms for light ion and neutron propagation with well defined convergence criteria in 3D objects is developed and tested against Monte Carlo simulations of 3D effects.

Space radiation accelerator experiments – The role of neutrons and light ions

Abstract: The importance of neutrons and light ions is considered when astronauts spend considerable time in thickly shielded regions of a spacecraft. This may be relevant for space missions both in and beyond low Earth orbit (LEO). In addition to heavy ion experiments at accelerators, it is suggested that an increased emphasis on experiments with lighter ions may be useful in reducing biological uncertainties.

Mitochondria regulate DNA damage and genomic instability induced by high LET radiation.

Abstract: High linear energy transfer (LET) radiation including α particles and heavy ions is the major type of radiation find in space and is considered a potential health risk for astronauts. Even though the chance that these high LET particles traversing through the cytoplasm of cells is higher than that through the nuclei, the contribution of targeted cytoplasmic irradiation, to the induction of genomic instability and other chromosomal damages induced by high LET radiation is not known. In the present study, we investigated whether mitochondria are the potential cytoplasmic target of high LET radiation in mediating cellular damage using a mitochondrial DNA (mtDNA) depleted (ρ0) human small airway epithelial (SAE) cell model and a precision charged particle microbeam with a beam width of merely one micron. Targeted cytoplasmic irradiation by high LET α particles induced DNA oxidative damage and double strand breaks in wild type ρ+ SAE cells. Furthermore, there was a significant increase in autophagy, micronuclei, which is an indication of genomic instability, together with the activation of nuclear factor kappa-B (NF-κB) and mitochondrial inducible nitric oxide synthase (iNOS) signaling pathways in ρ+ SAE cells. In contrast, ρ0 SAE cells exhibited a significantly lower response to these same endpoints examined after cytoplasmic irradiation with high LET α particles. The results indicate that mitochondria are essential in mediating cytoplasmic radiation induced genotoxic damage in mammalian cells. Furthermore, the findings may shed some light in the design of countermeasures for space radiation.

Acute hematological effects in mice exposed to the expected doses, dose-rates, and energies of solar particle event-like proton radiation

Abstract: The nature and assignment of the Martian spectral features observed by W. M. Sinton in the 3-4 micron region have been re-examined. It is shown that it is not possible to state unequivocally that there are absorption bands at 2710 (3.69 microns), 2790 (3.58 microns) and 2900 (3.45 microns) cm-1. Rather the possibility of the presence of true reflection features complicates the issue and enables only a wide range to be specified for the corresponding resonant frequencies. To clarify the assignment of these a large number of pure organic and inorganic solids, together with a selection of biological samples, have been recorded in reflection. The assignment of the features to organic matter, implying the presence of life on the Martian maria, is found to be improbable. While inorganic carbonates have bands in this region, they also do not provide a satisfactory explanation of the observations. The assignment of these Martian spectral features is then an open question.

56Fe Particle Exposure Results in a Long-Lasting Increase in a Cellular Index of Genomic Instability and Transiently Suppresses Adult Hippocampal Neurogenesis in Vivo.

Abstract: The high-LET HZE particles from galactic cosmic radiation pose tremendous health risks to astronauts, as they may incur sub-threshold brain injury or maladaptations that may lead to cognitive impairment. The health effects of HZE particles are difficult to predict and unfeasible to prevent. This underscores the importance of estimating radiation risks to the central nervous system as a whole as well as to specific brain regions like the hippocampus, which is central to learning and memory. Given that neurogenesis in the hippocampus has been linked to learning and memory, we investigated the response and recovery of neurogenesis and neural stem cells in the adult mouse hippocampal dentate gyrus after HZE particle exposure using two nestin transgenic reporter mouse lines to label and track radial glia stem cells (Nestin-GFP and Nestin-CreERT2/R26R:YFP mice, respectively). Mice were subjected to 56Fe particle exposure (0 or 1 Gy, at either 300 or 1000 MeV/n) and brains were harvested at early (24h), intermediate (7d), and/or long time points (2-3mo) post-irradiation. 56Fe particle exposure resulted in a robust increase in 53BP1+ foci at both the intermediate and long time points post-irradiation, suggesting long-term genomic instability in the brain. However, 56Fe particle exposure only produced a transient decrease in immature neuron number at the intermediate time point, with no significant decrease at the long time point post-irradiation. 56Fe particle exposure similarly produced a transient decrease in dividing progenitors, with fewer progenitors labeled at the early time point but equal number labeled at the intermediate time point, suggesting a recovery of neurogenesis. Notably, 56Fe particle exposure did not change the total number of nestin-expressing neural stem cells. These results highlight that despite the persistence of an index of genomic instability, 56Fe particle-induced deficits in adult hippocampal neurogenesis may be transient. These data support the regenerative capacity of the adult SGZ after HZE particle exposure and encourage additional inquiry into the relationship between radial glia stem cells and cognitive function after HZE particle exposure.

Are biological effects of space radiation really altered under the microgravity environment

Abstract: Two major factors of space environment are space radiation and microgravity. It is generally considered that a high level of ionizing radiation (IR) in space has an influence on living organisms including humans; therefore, the possible alteration of space-radiation influences by the microgravity environment is of great concern. In fact, examination of such a possibility has been extensively conducted since the early days of space experiments, suggesting a possible synergistic effect of radiation and microgravity in some experiments but a negative observation in others. Because these complicated results remain not well understood, we propose a solution to this problem. Gene expression analysis is one of the solutions to the problem. In fact, gene expression may be changed by microgravity, and further modification may be possible through IR. This result could reveal an interactive effect of both factors on the cellular responses, which could in turn reveal whether the human-health abnormalities expected under the microgravity environment can be altered by space radiation. We believe that this is a new aspect in the study of the interactive effect of radiation and microgravity. However, further improvements in space experimental technologies are required for future studies.

Effects of illuminants and illumination time on lettuce growth, yield and nutritional quality in a controlled environment

Abstract: Effects of illuminants and illumination time on the growth of lettuce were researched. Red–blue light-emitting diodes (LEDs, 90% red light +10% blue light) and white light fluorescent (WF) lamps were compared as the illuminants for plant cultivation. Under each type of illuminant, lettuce was grown at 4 illumination times: 12 h, 16 h, 20 h and 24 h, with the same light intensity of 600 μmol m − 2 s − 1 . The leaf net photosynthetic rate (Pn) under the two illuminants was comparable but the shape of lettuce was obviously affected by the illuminant. The WF lamps produced more compact plant, while red–blue LED resulted in less but longer leaves. However, the total leaf area was not significantly affected by the illuminant. The red–blue LED produced nearly same aboveground biomass with far less energy consumption relative to WF lamps. The underground biomass was lowered under red–blue LED in comparison with WF lamps. Red–blue LED could improve the nutritional quality of lettuce by increasing the concentration of soluble sugar and vitamin C ( V C ) and reducing the concentration of nitrate. Under each type of illuminant, longer illumination time resulted in higher Pn, more leaves and larger leaf area. The total chlorophyll concentration increased while the concentration ratio of chlorophyll a/b decreased with the extension of illumination time. Illumination time had highly significant positive correlation with biomass. Moreover, when total daily light input was kept the same, longer illumination time increased the biomass significantly as well. In addition, longer illumination time increased the concentration of crude fiber, soluble sugar and V C and reduced the concentration of nitrate. In summary, red–blue LEDs and 24 h illumination time were demonstrated to be more suitable for lettuce cultivation in the controlled environment.

Report of the workshop for life detection in samples from Mars

Abstract: The question of whether there is or was life on Mars has been one of the most pivotal since Schiaparellis' telescopic observations of the red planet. With the advent of the space age, this question can be addressed directly by exploring the surface of Mars and by bringing samples to Earth for analysis. The latter, however, is not free of problems. Life can be found virtually everywhere on Earth. Hence the potential for contaminating the Mars samples and compromising their scientific integrity is not negligible. Conversely, if life is present in samples from Mars, this may represent a potential source of extraterrestrial biological contamination for Earth. A range of measures and policies, collectively termed ‘planetary protection’, are employed to minimise risks and thereby prevent undesirable consequences for the terrestrial biosphere. This report documents discussions and conclusions from a workshop held in 2012, which followed a public conference focused on current capabilities for performing life-detection studies on Mars samples. The workshop focused on the evaluation of Mars samples that would maximise scientific productivity and inform decision making in the context of planetary protection. Workshop participants developed a strong consensus that the same measurements could be employed to effectively inform both science and planetary protection, when applied in the context of two competing hypotheses: 1) that there is no detectable life in the samples; or 2) that there is martian life in the samples. Participants then outlined a sequence for sample processing and defined analytical methods that would test these hypotheses. They also identified critical developments to enable the analysis of samples from Mars.

Changes in miRNA expression profile of space-flown Caenorhabditis elegans during Shenzhou-8 mission

Abstract: Recent advances in the field of molecular biology have demonstrated that small non-coding microRNAs (miRNAs) have a broad effect on gene expression networks and play a key role in biological responses to environmental stressors. However, little is known about how space radiation exposure and altered gravity affect miRNA expression. The "International Space Biological Experiments" project was carried out in November 2011 by an international collaboration between China and Germany during the Shenzhou-8 (SZ-8) mission. To study the effects of spaceflight on Caenorhabditis elegans (C. elegans), we explored the expression profile miRNA changes in space-flown C. elegans. Dauer C. elegans larvae were taken by SZ-8 spacecraft and experienced the 16.5-day shuttle spaceflight. We performed miRNA microarray analysis, and the results showed that 23 miRNAs were altered in a complex space environment and different expression patterns were observed in the space synthetic and radiation environments. Most putative target genes of the altered miRNAs in the space synthetic environment were predicted to be involved in developmental processes instead of in the regulation of transcription, and the enrichment of these genes was due to space radiation. Furthermore, integration analysis of the miRNA and mRNA expression profiles confirmed that twelve genes were differently regulated by seven miRNAs. These genes may be involved in embryonic development, reproduction, transcription factor activity, oviposition in a space synthetic environment, positive regulation of growth and body morphogenesis in a space radiation environment. Specifically, we found that cel-miR-52, -55, and -56 of the miR-51 family were sensitive to space environmental stressors and could regulate biological behavioural responses and neprilysin activity through the different isoforms of T01C4.1 and F18A12.8. These findings suggest that C. elegans responded to spaceflight by altering the expression of miRNAs and some target genes that function in diverse regulatory pathways.

Inhibition of microRNA-31-5p protects human colonic epithelial cells against ionizing radiation.

Abstract: MicroRNAs (miRNAs), endogenous non-coding small RNAs, are sensitive to environmental changes, and their differential expression is important for adaptation to the environment. However, application of miRNAs as a clinical prognostic or diagnostic tool remains unproven. In this study we demonstrate a chronic/persistent change of miRNAs from the plasma of a colorectal cancer susceptible mouse model (CPC;Apc) about 250 days after exposure to a simulated solar particle event (SPE). Differentially expressed miRNAs were identified compared to unirradiated control mice, including miR-31-5p, which we investigated further. To address the cellular function of miR-31-5p, we transfected a miR-31-5p mimic (sense) or inhibitor (antisense) into immortalized human colonic epithelial cells followed by gamma-irradiation. A miR-31-5p mimic sensitized but a miR-31-5p inhibitor protected colonic epithelial cells against radiation induced killing. We found that the miR-31-5p mimic inhibited the induction of hMLH1 expression after irradiation, whereas the miR-31-5p inhibitor increased the basal level of hMLH1 expression. The miR-31-5p inhibitor failed to modulate radiosensitivity in an hMLH1-deficient HCT116 colon cancer cell line but protected HCT116 3-6 and DLD-1 (both hMLH1-positive) colon cancer cell lines. Our findings demonstrate that miR-31-5p has an important role in radiation responses through regulation of hMLH1 expression. Targeting this pathway could be a promising therapeutic strategy for future personalized anti-cancer radiotherapy.

Quality factors for space radiation: A new approach.

Abstract: NASA has derived new models for radiological risk assessment based on epidemiological data and radiation biology including differences in Relative Biological Effectiveness for leukemia and solid tumors. Comprehensive approaches were used to develop new risk cross sections and the extension of these into recommendations for risk assessment during space missions. The methodology relies on published data generated and the extensive research initiative managed by the NASA Human Research Program (HRP) and reviewed by the National Academy of Sciences. This resulted in recommendations for revised specifications of quality factors, QNASA(Z,β) in terms of track structure concepts that extend beyond LET alone. The new paradigm for quality factors placed demands on radiation monitoring procedures that are not satisfied by existing dosimetry systems or particle spectrometers that are practical for space exploration where mass, volume, band width and power consumption are highly constrained. We have proposed a new definition of quality factors that relaxes the requirements for identifying charge, Z, and velocity, β, of the incident radiation while still preserving the functional form of the inherent risk functions. The departure from the exact description of QNASA(Z,β) is that the revised values are new functions of LET for solid cancers and leukemia. We present the motivation and process for developing the revised quality factors. We describe results of extensive simulations using GCR distributions in free space as well as the resulting spectra of primary and secondary particles behind aluminum shields and penetration through water. In all cases the revised dose averaged quality factors agreed with those based on the values obtained using QNASA(Z,β). This provides confidence that emerging technologies for space radiation dosimetry can provide real time measurements of dose and dose equivalent while satisfying constraints on size, mass, power and bandwidth. The revised quality factors are sufficiently generalized to be applicable to radiation protection practices beyond space exploration.

Evidence of Disseminated Intravascular Coagulation in a Porcine Model Following Radiation Exposure.

Abstract: Recent evidence has suggested that disseminated intravascular coagulation (DIC) plays an integral role in death at the LD 50 dose of either gamma or solar particle event (SPE)-like proton radiation in ferrets. In these studies, Yucatan minipigs were evaluated to determine whether they were susceptible to the development of radiation induced DIC. Yucatan minipigs were exposed to a dose of 2.5 Gray (Gy) with X-rays and monitored over the course of 30 days. Evidence of DIC was evaluated by way of thromboelastometry parameters, platelet counts, fibrinogen concentration, and the d-dimer assay. Pigs exposed to X-rays developed signs of DIC within 2 days' post-irradiation. The development of DIC was exacerbated over the course of the studies, and one of the pigs died at day 14 and another had to be euthanized on day 16 post-irradiation. For both of these pigs, DIC was evident at the time of death. The following observations were indicated or were suggestive of DIC: whole blood clotting was impaired (as evidenced by thromboelastometry alterations), there were decreased platelet counts, elevated d-dimer concentrations in the blood, and/or hemorrhaging and the presence of fibrin in tissues observed during post-mortem examination. The extrapolation of data from these studies, in combination with other published data, have led to the hypothesis that there could be a correlation between the propensity to develop DIC, as indicated by hemorrhaging at death at relatively low doses of radiation, and the LD 50 for a particular species. Our data suggest that the development of DIC may contribute to death at the LD 50 dose in large mammals.

Differential effects of p53 on bystander phenotypes induced by gamma ray and high LET heavy ion radiation.

Abstract: High LET particle irradiation has several potential advantages over γ-rays such as p53-independent response. The purpose of this work is to disclose the effect of p53 on the bystander effect induced by different LET irradiations and underlying mechanism. Lymphocyte cells of TK6 (wild type p53) and HMy2.CIR (mutated p53) were exposed to either low or high LET irradiation, then their mitochondrial dysfunction and ROS generation were detected. The micronuclei (MN) induction in HL-7702 hepatocytes co-cultured with irradiated lymphocytes was also measured. It was found that the mitochondrial dysfunction, p66(Shc) activation, and intracellular ROS were enhanced in TK6 but not in HMy2.CIR cells after γ-ray irradiation, but all of them were increased in both cell lines after carbon and iron irradiation. Consistently, the bystander effect of MN formation in HL-7702 cells was only triggered by γ-irradiated TK6 cells but not by γ-irradiated HMy2.CIR cells. But this bystander effect was induced by both lymphocyte cell lines after heavy ion irradiation. PFT-μ, an inhibitor of p53, only partly inhibited ROS generation and bystander effect induced by 30 keV/μm carbon-irradiated TK6 cells but failed to suppress the bystander effect induced by the TK6 cells irradiated with either 70 keV/μm carbon or 180 keV/μm iron. The mitochondrial inhibitors of rotenone and oligomycin eliminated heavy ion induced ROS generation in TK6 and HMy2.CIR cells and hence diminished the bystander effect on HL-7702 cells. These results clearly demonstrate that the bystander effect is p53-dependent for low LET irradiation, but it is p53-independent for high LET irradiation which may be because of p53-independent ROS generation due to mitochondrial dysfunction.

Comparison of space flight and heavy ion radiation induced genomic/epigenomic mutations in rice (Oryza sativa).

Abstract: Rice seeds, after space flight and low dose heavy ion radiation treatment were cultured on ground. Leaves of the mature plants were obtained for examination of genomic/epigenomic mutations by using amplified fragment length polymorphism (AFLP) and methylation sensitive amplification polymorphism (MSAP) method, respectively. The mutation sites were identified by fragment recovery and sequencing. The heritability of the mutations was detected in the next generation. Results showed that both space flight and low dose heavy ion radiation can induce significant alterations on rice genome and epigenome (P<0.05). For both genetic and epigenetic assays, while there was no significant difference in mutation rates and their ability to be inherited to the next generation, the site of mutations differed between the space flight and radiation treated groups. More than 50% of the mutation sites were shared by two radiation treated groups, radiated with different LET value and dose, while only about 20% of the mutation sites were shared by space flight group and radiation treated group. Moreover, in space flight group, we found that DNA methylation changes were more prone to occur on CNG sequence than CG sequence. Sequencing results proved that both space flight and heavy ion radiation induced mutations were widely spread on rice genome including coding region and repeated region. Our study described and compared the characters of space flight and low dose heavy ion radiation induced genomic/epigenomic mutations. Our data revealed the mechanisms of application of space environment for mutagenesis and crop breeding. Furthermore, this work implicated that the nature of mutations induced under space flight conditions may involve factors beyond ion radiation.

Effects of Fe particle irradiation on human endothelial barrier structure and function

Abstract: Space travel involves exposure to biologically effective heavy ion radiation and there is consequently a concern for possible degenerative disorders in humans. A significant target for radiation effects is the microvascular system, which is crucial to healthy functioning of the tissues. Its pathology is linked to disrupted endothelial barrier function and is not only a primary event in a range of degenerative diseases but also an important influencing factor in many others. Thus, an assessment of the effects of heavy ion radiation on endothelial barrier function would be useful for estimating the risks of space travel. This study was aimed at understanding the effects of high LET Fe particles (1 GeV/n) and is the first investigation of the effects of charged particles on the function of the human endothelial barrier. We used a set of established and novel endpoints to assess barrier function after exposure. These include, trans-endothelial electrical resistance (TEER), morphological effects, localization of adhesion and cell junction proteins (in 2D monolayers and in 3D tissue models), and permeability of molecules through the endothelial barrier. A dose of 0.50 Gy was sufficient to cause a progressive reduction in TEER measurements that were significant 48 hours after exposure. Concurrently, there were morphological changes and a 14% loss of cells from monolayers. Gaps also appeared in the normally continuous cell-border localization of the tight junction protein – ZO-1 but not the Platelet endothelial cell adhesion molecule (PECAM-1) in both monolayers and in 3D vessel models. Disruption of barrier function was confirmed by increased permeability to 3 kDa and 10 kDa dextran molecules. A dose of 0.25 Gy caused no detectible change in cell number, morphology, or TEER, but did cause barrier disruption since there were gaps in the cell border localization of ZO-1 and an increased permeability to 3 kDa dextran. These results indicate that Fe particles potently have impact on human endothelial barrier function and represent a risk for degenerative diseases in the space environment.

Response of cyanobacteria to low atmospheric pressure

Abstract: Maintaining a low pressure environment in a controlled ecological life support system would reduce the technological complexity and resupply cost in the course of the construction of a future manned lunar base. To estimate the effect of a hypobaric environment in a lunar base on biological components, such as higher plants, microbes, and algae, cyanobacteria was used as the model by determining their response of growth, morphology, and physiology when exposed to half of standard atmospheric pressure for 16 days (brought back to standard atmospheric pressure 30 minutes every two days for sampling). The results indicated that the decrease of atmospheric pressure from 100 kPa to 50 kPa reduced the growth rates of Microcystis aeruginosa , Merismopedia sp ., Anabaena sp . PCC 7120, and Anabaena flos-aquae . The ratio of carotenoid to chlorophyll a content in the four tested strains increased under low pressure conditions compared to ambient conditions, resulting from the decrease of chlorophyll a and the increase of carotenoid in the cells. Moreover, low pressure induced the reduction of the phycocyanin content in Microcystis aeruginosa , Anabaena sp . PCC 7120, and Anabaena flos-aquae . The result from the ultrastructure observed using SEM indicated that low pressure promoted the production of more extracellular polymeric substances (EPSs) compared to ambient conditions. The results implied that the low pressure environment of 50 kPa in a future lunar base would induce different effects on biological components in a CELSS, which must be considered during the course of designing a future lunar base. The results will be a reference for exploring the response of other biological components, such as plants, microbes, and animals, living in the life support system of a lunar base.

Biological system development for GraviSat: A new platform for studying photosynthesis and microalgae in space

Abstract: Microalgae have great potential to be used as part of a regenerative life support system and to facilitate in-situ resource utilization (ISRU) on long-duration human space missions. Little is currently known, however, about microalgal responses to the space environment over long (months) or even short (hours to days) time scales. We describe here the development of biological support subsystems for a prototype “3U” (i.e., three conjoined 10-cm cubes) nanosatellite, called GraviSat, designed to experimentally elucidate the effects of space microgravity and the radiation environment on microalgae and other microorganisms. The GraviSat project comprises the co-development of biological handling-and-support technologies with implementation of integrated measurement hardware for photosynthetic efficiency and physiological activity in support of long-duration (3–12 months) space missions. It supports sample replication in a fully autonomous system that will grow and analyze microalgal cultures in 120 μL wells around the circumference of a microfluidic polymer disc; the cultures will be launched while in stasis, then grown in orbit. The disc spins at different rotational velocities to generate a range of artificial gravity levels in space, from microgravity to multiples of Earth gravity. Development of the biological support technologies for GraviSat comprised the screening of more than twenty microalgal strains for various physical, metabolic and biochemical attributes that support prolonged growth in a microfluidic disc, as well as the capacity for reversible metabolic stasis. Hardware development included that necessary to facilitate accurate and precise measurements of physical parameters by optical methods (pulse amplitude modulated fluorometry) and electrochemical sensors (ion-sensitive microelectrodes). Nearly all microalgal strains were biocompatible with nanosatellite materials; however, microalgal growth was rapidly inhibited (∼1 week) within sealed microwells that did not include dissolved bicarbonate due to CO 2 starvation. Additionally, oxygen production by some microalgae resulted in bubble formation within the wells, which interfered with sensor measurements. Our research achieved prolonged growth periods (>10 months) without excess oxygen production using two microalgal strains, Chlorella vulgaris UTEX 29 and Dunaliella bardawil 30 861, by lowering light intensities (2–10 μmol photons m −2 s −1 ) and temperature (4–12 °C). Although the experiments described here were performed to develop the GraviSat platform, the results of this study should be useful for the incorporation of microalgae in other satellite payloads with low-volume microfluidic systems.

Orally administered fructose increases the numbers of peripheral lymphocytes reduced by exposure of mice to gamma or SPE-like proton radiation

Abstract: Exposure of the whole body or a major portion of the body to ionizing radiation can result in Acute Radiation Sickness (ARS), which can cause symptoms that range from mild to severe, and include death. One of the syndromes that can occur during ARS is the hematopoietic syndrome, which is characterized by a reduction in bone marrow cells as well as the number of circulating blood cells. Doses capable of causing this syndrome can result from conventional radiation therapy and accidental exposure to ionizing radiation. It is of concern that this syndrome could also occur during space exploration class missions in which astronauts could be exposed to significant doses of solar particle event (SPE) radiation. Of particular concern is the reduction of lymphocytes and granulocytes, which are major components of the immune system. A significant reduction in their numbers can compromise the immune system, causing a higher risk for the development of infections which could jeopardize the success of the mission. Although there are no specific countermeasures utilized for the ARS resulting from exposure to space radiation(s), granulocyte colony-stimulating factor (G-CSF) has been proposed as a countermeasure for the low number of neutrophils caused by SPE radiation, but so far no countermeasure exists for a reduced number of circulating lymphocytes. The present study demonstrates that orally administered fructose significantly increases the number of peripheral lymphocytes reduced by exposure of mice to 2 Gy of gamma- or SPE-like proton radiation, making it a potential countermeasure for this biological end-point.

Effects of hypobaria and hypoxia on seed germination of six plant species

Abstract: Hypobaria (low pressure) is typically associated with hypoxia (low oxygen partial pressure). There are several advantages of growing higher plants under hypobaria in the moon or mars habitat. The objectives of this research were to investigate the seed germination of six plant species under hypobaric and ambient total pressure conditions. Seeds were sown and germinated under three levels of total atmospheric pressure (101, 30 and 10 kPa) and three levels of oxygen partial pressures (21, 6 and 2 kPa) in an 8-day study. Hypoxia (6 or 2 kPa) significantly inhibited all seed germination under three levels of total atmospheric pressure by increasing the electrical conductivity and the optical density, decreasing the seed germination percentage and seed dehydrogenase activity and inhibiting the growth of the shoots and roots. Hypobaria (30 or 10 kPa) markedly improved seed germination and root growth by enhancing the oxygen diffusion rate under hypoxic conditions (6 or 2 kPa). The seeds of three dicot plants (lettuce, Chinese cabbage and cucumber) were more sensitive to hypoxia caused by hypobaria than were those of three monocot plants (maize, wheat and rice); lettuce and cucumber seeds had the highest sensitivity, whereas rice seeds had the lowest sensitivity. This research demonstrates that six experimental seeds can germinate normally under hypobaria (30 kPa), but the oxygen partial pressure should not be less than 6 kPa.

Space ethics to test directed panspermia

Abstract: The hypothesis that Earth was intentionally seeded with life by a preceding extraterrestrial civilization is believed to be currently untestable. However, analysis of the situation where humans themselves embark on seeding other planetary systems motivated by survival and propagation of life reveals at least two ethical issues calling for specific solutions. Assuming that generally intelligence evolves ethically as it evolves technologically, the same considerations might be applied to test the hypothesis of directed panspermia: if life on Earth was seeded intentionally, the two ethical requirements are expected to be satisfied, what appears to be the case.

Biological filter capable of simultaneous nitrification and denitrification for Aquatic Habitat in International Space Station.

Abstract: The biological filter capable of simultaneous nitrification and denitrification was constructed for aquatic animal experiments in the International Space Station (ISS). The biological filter will be used to remove harmful ammonia excreted from aquatic animals in a closed water circulation system (Aquatic Habitat). The biological filter is a cylindrical tank packed with porous glass beads for nitrification and dual plastic bags for denitrification. The porous beads are supporting media for Nitrosomonas europaea and Nitrobacter winogradskyi. The N. europaea cells and N. winogradskyi cells on the porous beads, oxidize the excreted ammonia to nitrate via nitrite. On the other hand, the dual bag is composed of an outer non-woven fabric bag and an inner non-porous polyethylene film bag. The outer bag is supporting media for Paracoccus pantotrophus. The inner bag, in which 99.5% ethanol is packed, releases the ethanol slowly, since ethanol can permeate through the non-porous polyethylene film. The P. pantotrophus cells on the outer bag reduce the produced nitrate to nitrogen gas by using the released ethanol as an electron donor for denitrification. The biological filter constructed in this study consequently removed the ammonia without accumulating nitrate. Most of the excess ethanol was consumed and did not affect the nitrification activity of the N. europaea cells and N. winogradskyi cells severely. In accordance with the aquatic animal experiments in the ISS, small freshwater fish had been bred in the closed water circulation system equipped with the biological filter for 90 days. Ammonia concentration daily excreted from fish is assumed to be 1.7 mg-N/L in the recirculation water. Under such conditions, the harmful ammonia and nitrite concentrations were kept below 0.1 mg-N/L in the recirculation water. Nitrate and total organic carbon concentrations in the recirculation water were kept below 5 mg-N/L and 3 mg-C/L, respectively. All breeding fish were alive and ate the feed well. The results show that the nitrification and denitrification abilities of the biological filter sufficed to keep water quality for aquatic animal experiments in the ISS. This simple and effective system is certainly applicable to aquarium systems and aquaculture systems.

Comparison of bone histomorphometry and μCT for evaluating bone quality in tail-suspended rats

Abstract: Astronauts often suffer from microgravity-induced osteoporosis due to their time in space. Bone histomorphometry, the ‘gold standard’ technique for detecting bone quality, is widely used in the evaluation of osteoporosis. This study investigates whether μCT has the same application value as histomorphometry in the evaluation of weightlessness-induced bone loss. A total of 24 SD rats were distributed into three groups ( n = 8 , each): tail-suspension (TS), TS plus active exercise (TSA), and control (CON). After 21 days, bone mineral density (BMD) was measured by dual energy X-ray absorptiometry (DXA) and μCT, and microstructure was measured by μCT and histomorphometry. BMD was found to have decreased significantly in TS and TSA compared with the CON group. The results of the μCT measurements showed that a change in BMD mainly occurred in the trabecular bone, and the trabecular BMD increased significantly in the TSA compared with the TS group. The comparison of μCT and histomorphometry showed that TS led to a significant decrease in bone volume (BV/TV), trabecular thickness (Tb.Th) and trabecular number (Tb.N), and it led to an increase in trabecular separation (Tb.Sp). However, active exercise can prevent these changes. Significant differences in most parameters between TSA and CON were found by μCT but not by histomorphometry. Additionally, the parameters of these two methods are highly correlated. Therefore, the application value of μCT is as good as histomorphometry and DXA in the diagnosis of weightlessness-induced osteoporosis and is even better in evaluating the efficacy of exercise.

Flexible imaging payload for real-time fluorescent biological imaging in parabolic, suborbital and space analog environments

Abstract: Fluorescent imaging offers the ability to monitor biological functions, in this case biological responses to space-related environments. For plants, fluorescent imaging can include general health indicators such as chlorophyll fluorescence as well as specific metabolic indicators such as engineered fluorescent reporters. This paper describes the Flex Imager a fluorescent imaging payload designed for Middeck Locker deployment and now tested on multiple flight and flight-related platforms. The Flex Imager and associated payload elements have been developed with a focus on ‘flexibility’ allowing for multiple imaging modalities and change-out of individual imaging or control components in the field. The imaging platform is contained within the standard Middeck Locker spaceflight form factor, with components affixed to a baseplate that permits easy rearrangement and fine adjustment of components. The Flex Imager utilizes standard software packages to simplify operation, operator training, and evaluation by flight provider flight test engineers, or by researchers processing the raw data. Images are obtained using a commercial cooled CCD image sensor, with light-emitting diodes for excitation and a suite of filters that allow biological samples to be imaged over wavelength bands of interest. Although baselined for the monitoring of green fluorescent protein and chlorophyll fluorescence from Arabidopsis samples, the Flex Imager payload permits imaging of any biological sample contained within a standard 10 cm by 10 cm square Petri plate. A sample holder was developed to secure sample plates under different flight profiles while permitting sample change-out should crewed operations be possible. In addition to crew-directed imaging, autonomous or telemetric operation of the payload is also a viable operational mode. An infrared camera has also been integrated into the Flex Imager payload to allow concurrent fluorescent and thermal imaging of samples. The Flex Imager has been utilized to assess, in real-time, the response of plants to novel environments including various spaceflight analogs, including several parabolic flight environments as well as hypobaric plant growth chambers. Basic performance results obtained under these operational environments, as well as laboratory-based tests are described. The Flex Imager has also been designed to be compatible with emerging suborbital platforms.

The evaluation of upper body muscle activity during the performance of external chest compressions in simulated hypogravity

Abstract: BACKGROUND: This original study evaluated the electromyograph (EMG) activity of four upper body muscles: triceps brachii, erector spinae, upper rectus abdominis, and pectoralis major, while external chest compressions (ECCs) were performed in simulated Martian hypogravity using a Body Suspension Device, counterweight system, and standard full body cardiopulmonary resuscitation (CPR) mannequin. METHOD: 20 young, healthy male subjects were recruited. One hundred compressions divided into four sets, with roughly six seconds between each set to indicate ‘ventilation’, were performed within approximately a 1.5 minute protocol. Chest compression rate, depth and number were measured along with the subject's heart rate (HR) and rating of perceived exertion (RPE). RESULTS: All mean values were used in two-tailed t-tests using SPSS to compare +1 Gz values (control) versus simulated hypogravity values. The AHA (2005) compression standards were maintained in hypogravity. RPE and HR increased by 32% ( p 0.001 ) and 44% ( p = 0.002 ), respectively, when ECCs were performed during Mars simulation, in comparison to +1 Gz. In hypogravity, the triceps brachii showed significantly less activity ( p 0.001 ) when compared with the other three muscles studied. The comparison of all the other muscles showed no difference at +1 Gz or in hypogravity. CONCLUSIONS: This study was among the first of its kind, however several limitations were faced which hopefully will not exist in future studies. Evaluation of a great number of muscles will allow space crews to focus on specific strengthening exercises within their current training regimes in case of a serious cardiac event in hypogravity.

Simulated weightlessness and synbiotic diet effects on rat bone mechanical strength

Abstract: This paper reports results on exposure to simulated weightlessness that leads to a rapid decrease in bone mineral density known as spaceflight osteopenia by evaluating the effectiveness of dietary supplementation with synbiotics to counteract the effects of skeletal unloading. Forty adult male rats were studied under four different conditions in a 2 × 2 factorial design with main effects of diet (synbiotic and control) and weight condition (unloaded and control). Hindlimb unloading was performed at all times for 14 days followed by 14 days of recovery (reambulation). The synbiotic diet contained probiotic strains Lactobacillus acidophilus and Lactococcus lactis lactis and prebiotic fructooligosaccharide. This paper also reports on the development of a desktop three-point bending device to measure the mechanical strength of bones from rats subjected to simulated weightlessness. The importance of quantifying bone resistance to breakage is critical when examining the effectiveness of interventions against osteopenia resulting from skeletal unloading, such as astronauts experience, disuse or disease. Mechanical strength indices provide information beyond measures of bone density and microarchitecture that enhance the overall assessment of a treatment's potency. In this study we used a newly constructed three-point bending device to measure the mechanical strength of femur and tibia bones from hindlimb-unloaded rats fed an experimental synbiotic diet enriched with probiotics and fermentable fiber. Two calculated outputs for each sample were Young's modulus of elasticity and fracture stress. Bone major elements (calcium, magnesium, and phosphorous) were quantified using ICP-MS analysis. Hindlimb unloading was associated with a significant loss of strength in the femur, and with significant reductions in major bone elements. The synbiotic diet did not protect against these unloading effects. Tibia strength and major elements were not reduced by hindlimb unloading, as was the case for femur, but tibia bone strength was negatively affected by the synbiotic diet. Thus, unexpectedly, the synbiotic diet was associated with null or detrimental effects on bone strength.

Proximity within interphase chromosome contributes to the breakpoint distribution in radiation-induced intrachromosomal exchanges

Abstract: Previously, we reported that breaks involved in chromosome aberrations were clustered in several regions of chromosome 3 in human mammary epithelial cells after exposures to either low- or high-LET radiation. In particular, breaks in certain regions of the chromosome tended to rejoin with each other to form an intrachromosome exchange event. This study tests the hypothesis that proximity within a single chromosome in interphase cell nuclei contributes to the distribution of radiation-induced chromosome breaks. Chromosome 3 in G1 human mammary epithelial cells was hybridized with the multicolor banding in situ hybridization (mBAND) probes that distinguish the chromosome in six differently colored regions, and the location of these regions was measured with a laser confocal microscope. Results of the study indicated that, on a multi-mega base pair scale of the DNA, the arrangement of chromatin was non-random. Both telomere regions tended to be located towards the exterior of the chromosome domain, whereas the centromere region towards the interior. In addition, the interior of the chromosome domain was preferentially occupied by the p-arm of the chromatin, which is consistent with our previous finding of intrachromosome exchanges involving breaks on the p-arm and in the centromere region of chromosome 3. Other factors, such as the fragile sites in the 3p21 band and gene regulation, may also contribute to the breakpoint distribution in radiation-induced chromosome aberrations.