Neurobiological problems in long-term deep space flights.

Future missions in space may involve long-term travel beyond the magnetic field of the Earth, subjecting astronauts to radiation hazards posed by solar flares and galactic cosmic rays, altered gravitation fields and physiological stress. Thus, it is critical to determine if there will be any reversible or irreversible, detrimental neurological effects from this prolonged exposure to space. A question of particular importance focuses on the long-term effects of the space environment on the central nervous system (CNS) neuroplasticity, with the potential acute and/or delayed effects that such perturbations might entail. Although the short-term effects of microgravity on neural control were studied on previous low earth orbit missions, the late consequences of stress in space, microgravity and space radiation have not been addressed sufficiently at the molecular, cellular and tissue levels. The possibility that space flight factors can interact influencing the neuroplastic response in the CNS looms critical issue not only to understand the ontogeny of the CNS and its functional integrity, but also, ultimately the performance of astronauts in extended space forays. The purpose of this paper is to review the neurobiological modifications that occur in the CNS exposed to the space environment, and its potential consequences for extended deep space flight.     

Effects of space flight and IGF-1 on immune function

We tested the hypothesis that insulin-like growth factor-1 (IGF-1) would ameliorate space flight-induced effects on the immune system. Twelve male, Sprague-Dawley rats, surgically implanted with mini osmotic pumps, were subjected to space flight for 10 days on STS-77. Six rats received 10 mg/kg/day of IGF-1 and 6 rats received saline. Flight animals had a lymphocytopenia and granulocytosis which were reversed by IGF-1. Flight animals had significantly higher corticosterone levels than ground controls but IGF-1 did not impact this stress hormone. Therefore, the reversed granulocytosis did not correlate with serum corticosterone. Space flight and IGF-1 also combined to induce a monocytopenia that was not evident in ground control animals treated with IGF-1 or in animals subjected to space flight but given physiological saline. There was a significant increase in spleen weights in vivarium animals treated with IGF-1, however, this change did not occur in flight animals. We observed reduced agonist-induced lymph node cell proliferation by cells from flight animals compared to ground controls. The reduced proliferation was not augmented by IGF-1 treatment. There was enhanced secretion of TNF, IL-6 and NO by flight-animal peritoneal macrophages compared to vivarium controls, however, O₂ secretion was not affected. These data suggest that IGF-1 can ameliorate some of the effects of space flight but that space flight can also impact the normal response to IGF-1.     

Ultrastructural and growth indices of Chlorella culture in multicomponent aquatic systems under space flight conditions.

Submicroscopic organization of Chlorella cells cultivated under space flight conditions in three-component aquatic system has been studied. Comparison of the experimental cells with that of the controls revealed certain rearrangements of cell organelles particularly, a reduction in the amount of reserve polysaccharides in chloroplasts, increase of cell vacuolization and mitochondrion volume, complication configuration of plasmalemma evaginations and invaginations, and also disturbances in the process of cytokinesis. More over an increase in the number of Chlorella cells infected by bacteria was shown in the experimental variant. No considerable differences were established in the growth characteristics of the experimental and control populations. A comparative cytological analysis revealed general regularities of organelles in Chlorella cells cultivated under space flight condition in the uni- and multicomponent systems.     

Accumulation of stress protein 72 (HSP72) in muscle and spleen of goldfish taken into space.

Using Western blot analysis, here, we report the levels of HSP72 in several organs from goldfish which were taken into space on the NASA space shuttle. A remarkable accumulation of HSP72 was detected in muscle and spleen of those fish taken into space as compared with controls. These results suggested that the HSP72 induction is a kind of stress response at the molecular level introduced by the space environment consisting of microgravity and/or cosmic radiation as stressors.     

Optimization of moisture content for wheat seedling germination in a cellulose acetate medium for a space flight experiment.

The Porous Tube Plant Nutrient Delivery System (PTPNDS), a hydrophilic, microporous ceramic tube hydroponic system designed for microgravity, will be tested in a middeck locker of the Space Shuttle. The flight experiment will focus on hardware operation and assess its ability to support seed germination and early seedling growth in microgravity. The water controlling system of the PTPNDS hardware has been successfully tested during the parabolic flight of the KC-135. One challenge to the development of the space flight experiment was to devise a method of holding seeds to the cylindrical porous tube. The seed-holder must provide water and air to the seed, absorb water from the porous tube, withstand sterilization, provide a clear path for shoots and roots to emerge, and be composed of flight qualified materials. In preparation for the flight experiment, a wheat seed-holder has been designed that utilizes a cellulose acetate plug to facilitate imbibition and to hold the wheat seeds in contact with the porous tube in the correct orientation during the vibration of launch and the microgravity environment of orbit. Germination and growth studies with wheat at a range of temperatures showed that optimal moisture was 78% (by weight) in the cellulose acetate seed holders. These and other design considerations are discussed.     

Induced abnormality in Mir- and Earth grown Super Dwarf wheat.

Super-dwarf wheat grown on the Mir space station using the Svet "Greenhouse" exhibited morphological, metabolic and reproductive abnormalities compared with Earth-grown wheat. Of prominent importance were the abnormalities associated with reproductive ontogeny and the total absence of seed formation on Mir. Changes in the apical meristem associated with transition from the vegetative phase to floral initiation and development of the reproductive spike were all typical of 'Super-Dwarf' wheat up to the point of anthesis. Observation of ruptured anthers from the Mir-grown plants revealed what appeared to be normally developed pollen. These pollen gains, however, contained only one nuclei, while normal viable pollen is tri-nucleate. A potentially important difference in the flight experiment, compared with ground reference studies, was the presence of a high level of atmospheric ethylene (1,200 ppb). Ground studies conducted by exposing 'Super-Dwarf' wheat to ethylene just prior to anthesis resulted in manifestation of the same abnormalities observed in the space flight samples.     

First flight of the ASTROCULTURE (TM) experiment as a part of the U.S. Shuttle/MIR program.

A number of space-based experiments have been conducted to assess the impact of microgravity on plant growth and development. In general, these experiments did not identify any profound impact of microgravity on plant growth and development, though investigations to study seed development have indicated difficulty in plants completing their reproductive cycle. However, it was not clear whether the lack of seed production was due to gravity effects or some other environmental condition prevailing in the unit used for conducting the experiment. The ASTROCULTURE (TM) flight unit contains a totally enclosed plant chamber in which all the critically important environmental conditions are controlled. Normal wheat (Triticum aestivum L.) growth and development in the ASTROCULTURE (TM) flight unit was observed during a ground experiment conducted prior to the space experiment. Subsequent to the ground experiment, the flight unit was transported to MIR by STS-89, as part of the U.S. Shuttle/MIR program, in an attempt to determine if super dwarf wheat plants that were germinated in microgravity would grow normally and produce seeds. The experiment was initiated on-orbit after the flight unit was transferred from the Space Shuttle to MIR. The ASTROCULTURE (TM) flight unit performed nominally for the first 24 hours after the flight unit was activated, and then the unit stopped functioning abruptly. Since it was not possible to return the unit to nominal operation it was decided to terminate the experiment. On return of the flight unit, it was confirmed that the control computer of the ASTROCULTURE (TM) flight unit sustained a radiation hit that affected the control software embedded in the computer. This experience points out that at high orbital inclinations, such as that of MIR and that projected for the International Space Station, the danger of encountering harmful radiation effects are likely unless the electronic components of the flight hardware are resistant to such impacts.     

Plant experiments with light-emitting diode module in Svet space greenhouse

Light is necessary for photosynthesis and shoot orientation in the space plant growth facilities. Light modules (LM) must provide sufficient photosynthetic photon flux for optimal efficiency of photosynthetic processes and also meet the constraints for power, volume and mass. A new LM for Svet space greenhouse using Cree® XLamp® 7090 XR light-emitting diodes (LEDs) was developed. Monochromic LEDs emitting in the red, green, and blue regions of the spectrum were used. The LED-LM contains 36 LED spots – 30 LED spots with one red, green and blue LED and 6 LED spots with three red LEDs. Digital Multiplex Control Unit controls the LED spots and can set 231 levels of light intensity thus achieving Photosynthetic Photon Flux Density (PPFD) in the range 0–400 μmol m⁻² s⁻¹ and different percentages of the red, green and blue light, depending on the experimental objectives. Two one-month experiments with plants – lettuce and radicchio were carried out at 400 μmol m⁻² s⁻¹ PPFD (high light – HL) and 220 μmol m⁻² s⁻¹ PPFD (low light – LL) and 70% red, 20% green and 10% blue light composition. To evaluate the efficiency of photosynthesis, in vivo modulated chlorophyll fluorescence was measured by Pulse Amplitude Modulation (PAM) fluorometer on leaf discs and the following parameters: effective quantum yield of Photosystem II (Φ_PSII) and non-photochemical quenching (NPQ) were calculated. Both lettuce and radicchio plants grown at LL express higher photochemical activity of Photosystem II (PSII) than HL grown plants, evaluated by Φ_PSII. Accelerated rise in NPQ in both LL grown plants was observed, while steady state NPQ values were higher in LL grown lettuce plants and did not differ in LL and HL grown radicchio plants. The extent of photoinhibition process in both plants was evaluated by changes in malonedialdehyde (MDA) concentration, peroxidase (POX) activity and hydrogen peroxide (H₂O₂) content. Accumulation of high levels of MDA and increased POX activity correlating with decreased H₂O₂ content were observed in both HL grown plants. These biochemical indicators revealed higher sensitivity to photodamage in HL grown lettuce and radicchio plants. LL conditions resulted in more effective functioning of PSII than HL when lettuce and radicchio plants were grown at 70% red, 20% green and 10% blue light composition.     

Seedling growth and development on space shuttle.

Young pine seedlings, and mung bean and oat seeds were flown on shuttle flights, STS-3 and STS-51F, in March, 1982 and July/August, 1985, respectively. The plant growth units built to support the two experiments functioned mechanically as anticipated and provided the necessary support data. Pine seedlings exposed to the microgravity environment of the space shuttle for 8 days continued to grow at a rate similar to ground controls. Pine stems in flight seedlings, however, averaged 10 to 12% less lignin than controls. Flight mung beans grew slower than control beans and their stems contained about 25% less lignin than control seedlings. Reduced mung bean growth in microgravity was partly due to slower germination rate. Lignin also was reduced in flight oats as compared to controls. Oats and mung beans exhibited upward growing roots which were not observed in control seedlings. Chlorophyll A/B ratios were lower in flight tissues than controls. The sealed PGCs exhibited large variations in atmospheric gas composition but the changes were similar between flight and ground controls. Ethylene was present in low concentrations in all chambers.     

The influence of space flight factors on viability and mutability of plants.

The experiments with air-dried Crepis capillaris seeds aboard the Soyuz 16 spaceship and the orbital stations Salyut 5, 6, 7 have revealed an increase in the frequency of aberrant cells in seedlings grown from flight-exposed seeds during the flight (experiment) and after the flight on Earth (flight control) as compared to the ground-based control. The increase in seedlings grown during the flight is more significant than in the flight control. During the flight Arabidopsis thaliana developed from cotyledons to the flowering stage. Analysis of seeds setting on these plants after the flight has shown a reduction in the fertility of these plants and an increase in the frequency of recessive mutants ("Light block-1"). An increased frequency of mutants was also retained in the progeny of plants which had passed through a complete cycle of development during the flight ("Fiton-3"). Suppression of embryo viability was observed in all experiments and expressed itself in reduced germinating ability of seeds from the exposed plants and in the early death of seedlings. Damages resulting from chromosome aberrations are eliminated in the first postflight generation and damages resulting from gene mutations and micro-aberrations are preserved for a longer time.     

Human reproductive issues in space.

Animal studies in space or analogous environments have suggested that there may be problems in the reproductive sphere; such factors might limit mankind's ability to live and work for extended periods of time in microgravity or on non-terrestrial planetary surfaces. A review of reproductive functioning in animal species studied during space flight demonstrated that most species were affected significantly by the absence of gravity and/or the presence of radiation. These two factors induced alterations in normal reproductive functioning independently of, as well as in combination with, each other. Based on animal models, we have identified several potential problem areas regarding human reproductive physiology and functioning in the space environment. While there are no current space flight investigations, the animal studies suggest priorities for future research in human reproduction. Such studies will be critical for the successful colonization of the space frontier.     

日地L1点探测任务设计与结果分析

日地L1点是太阳观测任务的理想观测点，对于中国后续太阳观测任务具有重要意义，因此在嫦娥五号的拓展任务阶段设计并实施了中国首次日地L1点探测任务，通过在轨飞行验证了日地L1点转移轨道、环绕轨道设计的正确性，对日地L1点的测控链路环境、太阳辐照环境、三体动力学环境、空间辐射环境等飞行环境进行了探测和验证。轨道飞行和各项环境探测的结果与设计模型的预示结果之间比对一致性较好，通过在轨飞行数据验证了设计模型的正确性。各项试验获得了预期的技术成果，进一步丰富了嫦娥五号任务的成果产出，对中国后续深空探测任务和产品的设计具有重要借鉴意义。     

The effect of microgravity on the development of plant protoplasts flown on Biokosmos 9.

An experiment using plant protoplasts has been accepted for the IML-1 Space Shuttle mission scheduled for 1991. Preparatory experiments have been performed using both fast and slow rotating clinostats and in orbit to study the effect of simulated and real weightlessness on protoplast regeneration. Late access to the space vehicles before launch has required special attention since it is important to delay cell wall regeneration until the samples are in orbit. On a flight on Biokosmos 9 ("Kosmos-2044") in September 1989 some preliminary results were obtained. Compared to the ground control, the growth of both carrot and rapeseed protoplasts was decreased by 18% and 44% respectively, after 14 days in orbit. The results also indicated that there is less cell wall regeneration under micro-g conditions. Compared to the ground controls the production of cellulose in rapeseed and carrot flight samples was only 46% and 29% respectively. The production of hemicellulose in the flight samples was 63% and 67% respectively of that of the ground controls. In both cases all samples reached the stage of callus development. The peroxidase activity was also found to be lower in the flight samples than in the ground controls, and the number of different isoenzymes was decreased in the flight samples. In general, the regeneration processes were retarded in the flight samples with respect to the ground controls. From a simulation experiment for IML-1 performed in January 1990 at ESTEC, Holland, regenerated plants have been obtained. These results are discussed and compared to the results obtained on Biokosmos 9. Protoplast regeneration did not develop beyond the callus stage in either the flight or the ground control samples from the Biokosmos 9 experiment.     

Protein expression profile changes in human fibroblasts induced by low dose energetic protons

Extrapolation of known radiation risks to the risks from low dose and low dose-rate exposures of human population, especially prolonged exposures of astronauts in the space radiation environment, relies in part on the mechanistic understanding of radiation induced biological consequences at the molecular level. While some genomic data at the mRNA level are available for cells or animals exposed to radiation, the data at the protein level are still lacking. Here, we studied protein expression profile changes using Panorama antibody microarray chips that contain antibodies to 224 proteins (or their phosphorylated forms) involved in cell signaling that included mostly apoptosis, cytoskeleton, cell cycle and signal transduction. Normal human fibroblasts were cultured until fully confluent and then exposed to 2 cGy of 150 MeV protons at high-dose rate. The proteins were isolated at 2 or 6 h after exposure and labeled with Cy3 for the irradiated cells and with Cy5 for the control samples before loading onto the protein microarray chips. The intensities of the protein spots were analyzed using ScanAlyze software and normalized by the summed fluorescence intensities and the housekeeping proteins. The results showed that low dose protons altered the expression of more than 10% of the proteins listed in the microarray analysis in various protein functional groups. Cell cycle (24%) related proteins were induced by protons and most of them were regulators of G1/S-transition phase. Comparison of the overall protein expression profiles, cell cycle related proteins, cytoskeleton and signal transduction protein groups showed significantly more changes induced by protons compared with other protein functional groups.     

Protein crystal growth aboard the U.S. space shuttle flights STS-31 and STS-32.

The first microgravity protein crystal growth experiments were performed on Spacelab I by Littke and John. These experiments indicated that the space grown crystals, which were obtained using a liquid-liquid diffusion system, were larger than crystals obtained by the same experimental system on earth. Subsequent experiments were performed by other investigators on a series of space shuttle missions from 1985 through 1990. The results from two of these shuttle flights (STS-26 and STS-29) have been described previously. The results from these missions indicated that the microgravity grown crystals for a number of different proteins were larger, displayed more uniform morphologies, and yielded diffraction data to significantly higher resolutions than the best crystals of these proteins grown on earth. This paper presents the results obtained from shuttle flight STS-32 (flown in January, 1990) and preliminary results from the most recent shuttle flight, STS-31 (flown in April, 1990).