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 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.     

The effects of simulated microgravity on the seminiferous tubules of rats

Space flight has been shown to have many adverse effects on various systems throughout the body. Because the opportunity to place research animals on board a Space Shuttle or the International Space Station is infrequent, various techniques have been designed to simulate the effects of microgravity in Earth based laboratories. A commonly used technique is known as antiorthostatic suspension, also often referred to as hind limb suspension. In this technique the hind portion of the animal is raised so that its hind limbs are non-weight bearing. This places the animal in roughly a 30° head down tilt position. This results in cephalic fluid shifts similar to those seen in actual space flight. This technique has also been shown to mimic other physiological parameters that are affected during space flight. This study examined testicular tissue from rats subjected to a 7 day antiorthostatic suspension. This tissue was acquired through a tissue sharing program and some of the experimental animals were injected with Interleukin 1 receptor antagonist (IL-1ra) which was hoped to ameliorate some of the effects of antiorthostatic suspension. The injection of IL-1ra was not expected to have any effect on testicular tissue, however this tissue was included in the morphological and statistical analysis to conduct a more complete study. All tissues were embedded in paraffin, sectioned, and stained using standard H&E staining. The tissue was then qualitatively ranked according to the “health” of the seminiferous tubules. Our findings indicate that 7 days of antiorthostatic suspension had adverse effects on the tissue that comprises the walls of the seminiferous tubules. It has long been known that antiorthostatic suspension has deleterious effects on testicular tissue, however this research indicates that these effects occur much faster than indicated by previous researchers. This is a significant finding because it indicates that meaningful earth based studies in this area can be carried out in a shorter time span. This could result in more studies per year as well as saving money by avoiding longer than necessary animal suspensions. This is especially important as we enter an era when, without Space Shuttle, flight opportunities will become scarce. These antiorthostatic suspension studies indicate that space flight, even short duration spaceflight, may have harmful effects on the seminiferous tubules and blood-testis barrier of astronauts.     

Production and action of cytokines in space.

B6MP102 cells, a continuously cultured murine bone marrow macrophage cell line, were tested for secretion of tumor necrosis factor-alpha and Interleukin-1 during space flight. We found that B6MP102 cells secreted more tumor necrosis factor-alpha and interleukin-1 when stimulated in space with lipopolysaccharide than controls similarly stimulated on earth. This compared to increased secretion of interferon-beta and -gamma by lymphocytes that was measured on the same shuttle flights. Although space flight enhanced B6MP102 secretion of tumor necrosis factor-alpha, an experiment on a subsequent space flight (STS-50) found that cellular cytotoxicity, mediated by tumor necrosis factor-alpha, was inhibited.     

Differentiation in microgravity of neural and muscle cells of a vertebrate (amphibian).

The CELIMENE space experiment (CELulles en Impesanteur: Muscle Et Neurone Embryonnaires) was devoted to the study of the influence of gravity on the differentiation, the organisation and the maintenance of the highly specialised nervous system and muscular system. CELIMENE was carried out during the first flight of the IBIS hardware (Instrument for BIology in Space) with the fully automatic space mission PHOTON 10 in February 1995. Using the amphibian Pleurodeles waltl as a vertebrate model, in vitro experiments involved immunocytochemical detection of glial-, neuronal- and muscle-specific markers, and neurotransmitters in cells developed under conditions of microgravity compared with 1g controls, on-board and on the ground. We observed that the altered gravity did not disturb cell morphogenesis or differentiation.     

The effects of space flight on some rat liver enzymes regulating carbohydrate and lipid metabolism

We have examined, in the livers of rats carried aboard the Cosmos 936 biosatellite, the activities of about 30 enzymes concerned with carbohydrate and lipid metabolism. In addition to the enzyme studies, the levels of glycogen and of the individual fatty acids in hepatic lipids were determined. Livers from flight and ground control rats at recovery (R₀) and 25 days after recovery (R₂₅) were used for these analyses.

For all parameters measured, the most meaningful comparisons are those made between flight stationary (FS) and flight centrifuged (FC) animals at R₀. When these two groups of flight rats were compared at R₀, statistically significant decreases in the activity levels of glycogen phosphorylase, -glycerol phosphate acyl transferase, diglyceride acyl transferase, aconitase and 6-phosphogluconate dehydrogenase and an increase in the palmitoyl CoA desaturase were noted in the weightless group (FS). The significance of these findings was strengthened by the fact that all enzyme activities showing alterations at R₀ returned to normal 25 days postflight. When liver glycogen and total fatty acids of the two sets of flight animals were determined, significant differences that could be attributed to reduced gravity were observed. The weightless group (FS) at R₀ contained, on the average, more than twice the amount of glycogen than did the centrifuged controls (FC) and a remarkable shift in the ratio of palmitate to palmitoleate was noted. These metabolic alterations, both in enzyme levels and in hepatic constituents, appear to be characteristic of the weightless condition. Our data seem to justify the conclusion that centrifugation during flight is equivalent to terrestrial gravity.     

The effect of the space flight environment on mucin production in the mouse uterine tube

Numerous studies have indicated that the microgravity environment of space has harmful effects on several tissues throughout the body. Although this phenomenon is well documented, research in this area is still in its relative infancy. This study investigates the effects of space flight on mucin production of the uterine tubes of mice. This study examined the epithelium of the uterine tubes from female mice that were flown on the space shuttle Endeavour for 13 days in August, 2007 and their concomitant controls. The tissue was qualitatively analyzed for the type of mucin produced, i.e., acidic, neutral, acidic/neutral mixture. Further, the tissue was quantitatively analyzed for the amounts of mucins produced by measuring the thickness of the mucin layer for each region of the uterine tube: isthmus, ampulla, and infundibulum. One way ANOVA tests were used to compare mucin thickness between all three sets of animals. Results indicate similar but not identical results between the three regions of the uterine tube. The Baseline tissue had the thickest mucin layer regardless of treatment group. In the ampulla the mucin layer was the thinnest in the Flight tissue, followed by the Ground Control, with the Baseline being the thickest. Analysis of the mucin layer of the infundibulum of the three treatment groups indicated no difference in its thickness between the three regions of the uterine tube. These results indicate a trend toward thinning of the mucin layer of the uterine tube in space flight, but also indicate an influence by the housing environment.     

Effect of the space environment on the induction of DNA-repair related proteins and recovery from radiation damage.

Recovery of bacterial cells from radiation damage and the effects of microgravity were examined in an STS-79 Shuttle/Mir Mission-4 experiment using the extremely radioresistant bacterium Deinococcus radiodurans. The cells were irradiated with gamma rays before the space flight and incubated on board the Space-Shuttle. The survival of the wild type cells incubated in space increased compared with the ground controls, suggesting that the recovery of this bacterium from radiation damage was enhanced under microgravity. No difference was observed for the survival of radiosensitive mutant rec30 cells whether incubated in space or on the ground. The amount of DNA-repair related RecA protein induced under microgravity was similar to those of ground controls, however, induction of PprA protein, the product of a newly found gene related to the DNA repair mechanism of D. radiodurans, was enhanced under microgravity compared with ground controls.     

Activity of the sympathetic-adrenomedullary system in rats after space flight on the COSMOS biosatellites

The indicators of adrenomedullary activity (catecholamine content (CA) and the activity of the catecholamine-synthesizing enzymes tyrosine hydroxylase (TH) and dopamine-β-hydroxylase (DBH)) were measured in the adrenal glands of rats living in a state of weightlessness for 18.5–19.5 days on board the biosatellites COSMOS 936 and COSMOS 1129. None of these indicators was significantly changed by space flight, neither in the group living in a state of weightlessness nor in the group living in a centrifuge on board the spacecraft and exposed to artificial gravity of 1 g (COSMOS 936). Animals exposed after space flight to repeated immobilization stress on Earth showed a significant decrease of adrenal adrenaline and an appreciable increase in adrenal TH activity compared to stressed animals which were not in space. These results suggest that a prolonged state of weightlessness during space flight does not by itself represent an intensive stressful stimulus for the adrenomedullary system but potentiates the response of cosmonauts to stress after return to Earth.     

Reduced lymphocyte activation in space: role of cell-substratum interactions.

We investigated the effect of substratum adhesiveness on stimulated lymphocyte blastogenesis by reducing and blocking cell adhesion with poly (2-hydroxyethyl methacrylate) (poly-HEMA) in a simple on-ground system. Cells grown on medium-thick and thick poly-HEMA films were rounded in shape and displayed no signs of spreading. By contrast, on tissue culture plastic and very thin poly-HEMA films, they showed clear signs of spreading. The mitogenic response of lymphocytes grown on thick poly-HEMA films was reduced by up to 68% of the control (tissue culture plastic). Interferon-gamma production was near zero when the cells were grown on the least adhesive substratum. On uncoated plastic, activated lymphocytes subjected to high gravity (20g) exhibited an increased proliferation rate (40%) compared with 1g. By contrast, on poly-HEMA, high gravity did not improve lymphocyte responsiveness. These results show that activated lymphocytes need to anchor and spread prior to achieving an optimal proliferation response. We conclude that decreased lymphocyte adhesion could contribute to the depressed in vitro lymphocyte responsiveness found in the microgravity conditions of space flight.     

空间小鼠实验有效载荷研究进展

啮齿动物是空间研究中常见的模型对象，已开展的空间生物实验中，小鼠表现出对飞行条件的有效适应.通过空间培养，研究者可对失重环境下小鼠的生理行为、骨骼和神经系统变化做进一步研究分析.本文对空间小鼠实验有效载荷的研究进展及其空间飞行实验进行了概述，为中国空间站上的小鼠培养箱设计提供参考.概述了地面小鼠培养技术和装置，为空间小鼠实验有效载荷设计提供地面对照；从实验研究内容和硬件系统设计两方面介绍了国际上已开展的空间小鼠实验，为中国空间生命科学研究和工程研制提供参考；对于中国发展空间小鼠实验有效载荷提出了建议.      

Possible mechanism of microgravity impact on Carausius morosus ontogenesis.

Experiments aboard "Spacelab-D1" and "Cosmos-1887" revealed an adverse effect of space flight on Carausius morosus embryos. The main influencing factor for stick insect eggs turned out to be microgravity, while the contribution of HZE particles of cosmic radiation was relatively low. Flight experiments indicated an increased vulnerability of stick insect eggs to microgravity at intermediate stages of development, that could support the "convection" hypothesis.     

Effects of space flight on surface marker expression.

Space flight has been shown to affect expression of several cell surface markers. These markers play important roles in regulation of immune responses, including CD4 and CD8. The studies have involved flight of experimental animals and humans followed by analysis of tissue samples (blood in humans, rats and monkeys, spleen, thymus, lymph nodes and bone marrow in rodents). The degree and direction of the changes induced by space flight have been determined by the conditions of the flight. Also, there may be compartmentalization of the response of surface markers to space flight, with differences in the response of cells isolated from blood and local immune tissue. The same type of compartmentalization was also observed with cell adhesion molecules (integrins). In this case, the expression of integrins from lymph node cells differed from that of splenocytes isolated from rats immediately after space flight. Cell culture studies have indicated that there may be an inhibition in conversion of a precursor cell line to cells exhibiting mature macrophage characteristics after space flight, however, these experiments were limited as a result of technical difficulties. In general, it is clear that space flight results in alterations of cell surface markers. The biological significance of these changes remains to be established.     

Development overview of the revised NASA Ultra Long Duration Balloon

The desire for longer duration stratospheric flights at constant float altitudes for heavy payloads has been the focus of the development of the National Aeronautics and Space Administration’s (NASA) Ultra Long Duration Balloon (ULDB) effort. Recent efforts have focused on ground testing and analysis to understand the previously observed issue of balloon deployment. A revised approach to the pumpkin balloon design has been tested through ground testing of model balloons and through two test flights. The design approach does not require foreshortening, and will significantly reduce the balloon handling during manufacture reducing the chances of inducing damage to the envelope. Successful ground testing of model balloons lead to the fabrication and test flight of a ∼176,000 m³ (∼6.2 MCF – Million Cubic Foot) balloon. Pre-flight analytical predictions predicted that the proposed flight balloon design to be stable and should fully deploy. This paper provides an overview of this first test flight of the revised Ultra Long Duration Balloon design which was a short domestic test flight from Ft. Sumner, NM, USA. This balloon fully deployed, but developed a leak under pressurization. After an extensive investigation to the cause of the leak, a second test flight balloon was fabricated. This ∼176,000 m³ (∼6.2 MCF) balloon was flown from Kiruna, Sweden in June of 2006. Flight results for both test flights, including flight performance are presented.     

Structure of cress root statocytes in microgravity (Bion-10 mission).

Experiments on primary roots of Lepidium sativum L. have been performed on board the Bion-10 satellite. The experimental set-up was extremely miniaturized and completely automatic. The results demonstrate the effectiveness of the instrumentation. The spatial orientation, growth, root cap differentiation and statocyte structure of roots grown under microgravity (MG) have been compared with control roots grown on the ground (GC) and in a 1G-reference centrifuge in space (RC). Root length and cap shape did not differ between MG and control samples. Under MG, the mean distance of the statoliths from the distal cell wall of the statocytes increased significantly, the mean distance of the mitochondria decreased and the nucleus did not change its position in comparison to both controls. The number and the shape of the amyloplasts (statoliths) were not influenced by the space flight factors, but their size as well as their relative area in the cell decreased. The number of starch grains per statolith as well as their size and shape changed under MG. In MG and RC samples the number of lipid bodies in the statocytes was higher and the relative area larger than in GC samples. The relative area occupied by vacuoles was greater in RC statocytes than in GC and MG statocytes. These results partly confirm and, in addition, extend the data from earlier experiments in space.     

Composition and physical properties of starch in microgravity-grown plants.

The effect of spaceflight on starch development in soybean (Glycine max L., BRIC-03) and potato (Solanum tuberosum, Astroculture-05) was compared with ground controls by biophysical and biochemical measurements. Starch grains from plants from both flights were on average 20-50% smaller in diameter than ground controls. The ratio delta X/delta rho (delta X --difference of magnetic susceptibilities, delta rho--difference of densities between starch and water) of starch grains was ca. 15% and 4% higher for space-grown soybean cotyledons and potato tubers, respectively, than in corresponding ground controls. Since the densities of particles were similar for all samples (1.36 to 1.38 g/cm3), the observed difference in delta X/delta rho was due to different magnetic susceptibilities and indicates modified composition of starch grains. In starch preparations from soybean cotyledons (BRIC-03) subjected to controlled enzymatic degradation with alpha-amylase for 24 hours, 77 +/- 6% of the starch from the flight cotyledons was degraded compared to 58 +/- 12% in ground controls. The amylose content in starch was also higher in space-grown tissues. The good correlation between the amylose content and delta X/delta rho suggests, that the magnetic susceptibility of starch grains is related to their amylose content. Since the seedlings from the BRIC-03 experiment showed elevated post-flight ethylene levels, material from another flight experiment (GENEX) which had normal levels of ethylene was examined and showed no difference to ground controls in size distribution, density, delta X/delta rho and amylose content. Therefore the role of ethylene appears to be more important for changes in starch metabolism than microgravity.     

Unique radiobiological aspects of high-LET radiation.

Since the beg inning of manned space flight the potentially unique radiobiological properties of the heavy ions of the cosmic radiation had been, apart from possible interactions of radiation effects with biological effects of weightlessness, of major concern with respect to the assessment of radiation hazards in manned space flight. Radiobiological findings obtained from space flight experiments and ground based experiments with densely ionizing radiation are discussed, which suggest qualitative differences between the radiobiological mechanisms of sparsely ionizing and densely ionizing radiation. These findings comprise the observation of a long lateral range of radiobiological effectiveness around tracks of single heavy ions, the observation of micro lesions induced in biological targets by the penetration of heavy ions, the nonadditivity of radiobiological effects from sparsely and densely ionizing radiation, the different kinetics for the expression of late effects induced by sparsely or densely ionizing radiation, and the observation of a reversed dose rate effect for early and late effects induced by densely ionizing radiation. These findings bear on the radiation protection standards to be installed for a general public in manned space flight and on the design of experiments, which intend to contribute to their specification.     

Animal models for the study of the effects of spaceflight on the immune system.

Animal models have been used to determine the effects of spaceflight on the immune system. Rats and rhesus monkeys have been the primary animals used for actual space flight studies, but mice have also been utilized for studies in ground-based models. The primary ground based model used has been hindlimb unloading of rodents, which is similar to the chronic bed-rest model for humans. A variety of immune responses have been shown to be modified when animals are hindlimb unloaded. These results parallel those observed when animals are flown in space. In general, immune responses are depressed in animals maintained in the hindlimb unloading model or flown in space. These results raise the possibility that spaceflight could result in decreased resistance to infection in animals.     

Space flight, microgravity, stress, and immune responses.

Exposure of animals and humans to space flight conditions has resulted in numerous alterations in immunological parameters. Decreases in lymphocyte blastogenesis, cytokine production, and natural killer cell activity have all been reported after space flight. Alterations in leukocyte subset distribution have also been reported after flight of humans and animals in space. The relative contribution of microgravity conditions and stress to the observed results has not been established. Antiorthostatic, hypokinetic, hypodynamic, suspension of rodents and chronic head-down tilt bed-rest of humans have been used to model effects of microgravity on immune responses. After use of these models, some effects of space flight on immune responses, such as decreases in cytokine function, were observed, but others, such as alterations in leukocyte subset distribution, were not observed. These results suggest that stresses that occur during space flight could combine with microgravity conditions in inducing the changes seen in immune responses after space flight. The biological/biomedical significance of space flight induced changes in immune parameters remains to be established. Grant Numbers: NCC2-859, NAG2-933.