Reproduction during spaceflight by plants in the family Brassicaceae

Researchers report on studies of reproduction in Arabidopsis thaliana in space during during the Chromex-03 on STS-54, Chromex-04 on STS-51, and Chromex-05 on STS-68 missions. The obstacles to seed formation were related to carbon dioxide levels. Other experiments examined in flight pollination and seed production in Brassica rapa during parabolic flight, a 4-1/2 month stay on Mir, and on STS-87. During the Mir experiment, Brassica seeds were harvested from seeds sown in flight. The second generation seeds grew to produce new seeds that contained more starch and less protein and lipid when compared to ground control seeds.     

Some aspects of the comparative analysis of hemodynamic responses to LBNP tests in cosmonauts of different age groups

The age-specific indicators of the functions of the cardiovascular system and its responses to the lower body negative pressure (LBNP) test were studied in career cosmonauts for the first time. The results of 174 LBNP tests implemented within the standard medical monitoring program on board the ??Mir?? orbital station (OS ??Mir??), using a Gamma-01 device, and aboard the International Space Station (ISS), using the Gamma-1M complex, were subjected to comparative analysis. In total, 38 cosmonauts from 25 long-duration space missions on board the ??Mir?? OS and ISS, who were examined in their pre-flight state and during in-flight periods, beginning, typically, on flight day (FD) 120, were subdivided into two age groups: 30- to 39-year-olds (their mean age was 36 ± 0.7 years; 39% of the total number of subjects) and 40- to 55-year olds (their mean age was 46 ± 0.8 years; 61% of the total number of subjects). We have revealed age-specific indicators for the hemodynamic status recorded at each stage of the investigation: at rest in a preflight state; responses of the indicators to the effects of microgravity; the relative dynamics of the indicators due to a simulated orthostatic posture, which was unidirectional but substantially different at the pre-and in-flight stages. For purposes of medical control, our results have shown that we need to establish age-specific references in our methodical approaches to the analysis and interpretation of the data received from monitoring cosmonauts?? health in their preflight state and during the entire mission and, which is particularly important in practical terms, when evaluating the LBNP test intolerance at different flight stages.     

Cytogenetic studies of blood lymphocytes of cosmonauts after long-ter, space flights

An analysis was performed of unstable chromosomal aberrations in peripheral blood of 36 cosmonauts after long-term space missions on "Mir" orbital station. 25 cosmonauts were examined before their flights to score spontaneous yields of cytogenetical damage. In all cases the doses absorbed by crews during space flights did not exceed permissible levels of irradiation, adopted for cosmonauts. The frequencies of chromosomal-type aberrations after space missions were found to increase significantly compared to the pre-flight levels. The yields of dicentrics and centric rings on the average were as high as 0.12 +/- 0.02 and 0.47 +/- 0.06% before and after the 1st flight, 0.18 +/- 0.05 and 0.71 +/- 0.11% before and after the 2nd flight respectively. During the inter-flight periods, usually lasted 1.5-2 years, the yields of chromosome damage lowered, but did not reach their spontaneous values. After each next flight the yields of chromosome aberrations increased again. The cytogenetical damage detected in cosmonauts' peripheral blood lymphocytes after chronic action of low doses of space radiation points out a possible increase in risks of stochastic effects in distant future for crews after long-term space missions.     

Influence of microgravity on ultrastructure and storage reserves in seeds of Brassica rapa L.

Successful plant reproduction under spaceflight conditions has been problematic in the past. During a 122 d opportunity on the Mir space station, full life cycles of Brassica rapa L. were completed in microgravity in a series of three experiments in the Svet greenhouse. Ultrastructural and cytochemical analyses of storage reserves in mature dry seeds produced in these experiments were compared with those of seeds produced during a high-fidelity ground control. Additional analyses were performed on developing Brassica embryos, 15 d post pollination, which were produced during a separate experiment on the Shuttle (STS-87). Seeds produced on Mir had less than 20% of the cotyledon cell number found in seeds harvested from the ground control. Cytochemical localization of storage reserves in mature cotyledons showed that starch was retained in the spaceflight material, whereas protein and lipid were the primary storage reserves in ground control seeds. Protein bodies in mature cotyledons produced in space were 44% smaller than those in the ground control seeds. Fifteen days after pollination, cotyledon cells from mature embryos formed in space had large numbers of starch grains, and protein bodies were absent, while in developing ground control seeds at the same stage, protein bodies had already formed and fewer starch grains were evident. These data suggest that both the late stage of seed development and maturation are changed in Brassica by growth in a microgravity environment. While gravity is not absolutely required for any step in the plant life cycle, seed quality in Brassica is compromised by development in microgravity.     

Analysis of deletion mutations of the rpsL gene in the yeast Saccharomyces cerevisiae detected after long-term flight on the Russian space station Mir

Using the yeast Saccharomyces cerevisiae on board the Russian space station Mir, we studied the effects of long-term space flight on mutation of the bacterial ribosomal protein L gene (rpsL) cloned in a yeast-Escherichia coli shuttle vector. The mutation frequencies of the cloned rpsL gene on the Mir and the ground (control) yeast samples were estimated by transformation of E. coli with the plasmid DNAs recovered from yeast and by assessment of the conversion of the rpsL wild-type phenotype (Sm(S)) to its mutant phenotype (Sm(R)). After a 40-day space flight, some part of space samples gave mutation frequencies two to three times higher than those of the ground samples. Nucleotide sequence analysis showed no apparent difference in point mutation rates between the space and the ground mutant samples. However, the greater part of the Mir mutant samples were found to have a total or large deletion in the rpsL sequence, suggesting that space radiation containing high-linear energy transfer (LET) might have caused deletion-type mutations.     

Muscle volume, MRI relaxation times (T2), and body composition after spaceflight.

Postflight changes in muscle volume, calf muscle transverse relaxation time, and total body composition were measured in 4 crewmembers after a 17-day mission and in 14-16 crewmembers in multiple shuttle/Mir missions of 16- to 28-wk duration. During the 17-day mission, all muscle regions except the hamstrings significantly decreased 3-10% compared with baseline. During the shuttle/Mir missions, there were significant decreases in muscle volume (5-17%) in all muscle groups except the neck. These changes, which reached a new steady state by 4 mo of flight or less, were reversed within 30-60 days after landing. Postflight swelling and elevation of calf muscle transverse relaxation time persisted for several weeks after flight, which suggests possible muscle damage. In contrast to the 17-day flight, in which loss in fat, but not lean body mass, was found (25), losses in bone mineral content and lean body mass, but not fat, were seen after the longer shuttle/Mir missions. The percent losses in total body lean body mass and bone mineral content were similar at approximately 3.4-3.5%, whereas the pelvis demonstrated the largest regional bone loss at 13%.     

Effects of microgravity on the larval development, metamorphosis and reproduction of the urodele amphibian Pleurodeles waltl

The FERTILE experiment was twice performed onboard the Mir space station during the Cassiopée and Pégase French space missions. The goal was to analyze the effects of microgravity on fertilization and embryonic development, and then on further development on the ground in the amphibian Pleurodeles waltl. The present paper reports development that occurred in the laboratory after landing. Recovered on the ground at the hatching stage, young larvae reared at room temperature underwent metamorphosis and became adults without obvious abnormalities. Of particular interest was the rearing temperature that induced a delayed metamorphosis for animals from the Cassiopée space mission, but not for animals from the Pégase mission. The rate of development and the morphology were analogous in these animals and in ground controls reared in a similar annual period. Analysis of offspring was performed using these animals. Males born in space were first mated with control ground-born females and then with females born in space. The mating gave progeny that developed normally. Depending on the methods used and on the limits of the analyses, the results clearly demonstrated that animals born in space were able to live and reproduce after return to the ground.     

Nine months in space: effects on human autonomic cardiovascular regulation.

We studied three Russian cosmonauts to better understand how long-term exposure to microgravity affects autonomic cardiovascular control. We recorded the electrocardiogram, finger photoplethysmographic pressure, and respiratory flow before, during, and after two 9-mo missions to the Russian space station Mir. Measurements were made during four modes of breathing: 1) uncontrolled spontaneous breathing; 2) stepwise breathing at six different frequencies; 3) fixed-frequency breathing; and 4) random-frequency breathing. R wave-to-R wave (R-R) interval standard deviations decreased in all and respiratory frequency R-R interval spectral power decreased in two cosmonauts in space. Two weeks after the cosmonauts returned to Earth, R-R interval spectral power was decreased, and systolic pressure spectral power was increased in all. The transfer function between systolic pressures and R-R intervals was reduced in-flight, was reduced further the day after landing, and had not returned to preflight levels by 14 days after landing. Our results suggest that long-duration spaceflight reduces vagal-cardiac nerve traffic and decreases vagal baroreflex gain and that these changes may persist as long as 2 wk after return to Earth.     

Comparative floral development of Mir-grown and ethylene-treated,earth-grown Super Dwarf wheat

To study plant growth in microgravity, we grew Super Dwarf wheat (Triticum aestivum L.) in the Svet growth chamber onboard the orbiting Russian space station, Mir, and in identical ground control units at the Institute of BioMedical Problems in Moscow, Russia. Seedling emergence was 56% and 73% in the two root-module compartments on Mir and 75% and 90% on earth. Growth was vigorous (produced ca. 1 kg dry mass), and individual plants produced 5 to 8 tillers on Mir compared with 3 to 5 on earth-grown controls. Upon harvest in space and return to earth, however, all inflorescences of the flight-grown plants were sterile. To ascertain if Super Dwarf wheat responded to the 1.1 to 1.7 micromoles mol-1 atmospheric levels of ethylene measured on the Mir prior to and during flowering, plants on earth were exposed to 0, 1, 3, 10, and 20 micromoles mol-1 of ethylene gas and 1200 micromoles mol-1 CO2 from 7 d after emergence to maturity. As in our Mir wheat, plant height, awn length, and the flag leaf were significantly shorter in the ethylene-exposed plants than in controls; inflorescences also exhibited 100% sterility. Scanning-electron-microscopic (SEM) examination of florets from Mir-grown and ethylene-treated, earth-grown plants showed that development ceased prior to anthesis, and the anthers did not dehisce. Laser scanning confocal microscopic (LSCM) examination of pollen grains from Mir and ethylene-treated plants on earth exhibited zero, one, and occasionally two, but rarely three nuclei; pollen produced in the absence of ethylene was always trinucleate, the normal condition. The scarcity of trinucleate pollen, abrupt cessation of floret development prior to anthesis, and excess tillering in wheat plants on Mir and in ethylene-containing atmospheres on earth build a strong case for the ethylene on Mir as the agent for the induced male sterility and other symptoms, rather than microgravity.     

Spaceflight Promotes Biofilm Formation by Pseudomonas aeruginosa

Understanding the effects of spaceflight on microbial communities is crucial for the success of long-term, manned space missions. Surface-associated bacterial communities, known as biofilms, were abundant on the Mir space station and continue to be a challenge on the International Space Station. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of immune function observed during spaceflight. While planktonic cultures of microbes have indicated that spaceflight can lead to increases in growth and virulence, the effects of spaceflight on biofilm development and physiology remain unclear. To address this issue, Pseudomonas aeruginosa was cultured during two Space Shuttle Atlantis missions: STS-132 and STS-135, and the biofilms formed during spaceflight were characterized. Spaceflight was observed to increase the number of viable cells, biofilm biomass, and thickness relative to normal gravity controls. Moreover, the biofilms formed during spaceflight exhibited a column-and-canopy structure that has not been observed on Earth. The increase in the amount of biofilms and the formation of the novel architecture during spaceflight were observed to be independent of carbon source and phosphate concentrations in the media. However, flagella-driven motility was shown to be essential for the formation of this biofilm architecture during spaceflight. These findings represent the first evidence that spaceflight affects community-level behaviors of bacteria and highlight the importance of understanding how both harmful and beneficial human-microbe interactions may be altered during spaceflight.     

Analysis of deletion mutations of the rpsL gene in the yeast Saccharomyces cerevisiae detected after long-term flight on the Russian space station Mir

Using the yeast Saccharomyces cerevisiae on board the Russian space station Mir, we studied the effects of long-term space flight on mutation of the bacterial ribosomal protein L gene (rpsL) cloned in a yeast-Escherichia coli shuttle vector. The mutation frequencies of the cloned rpsL gene on the Mir and the ground (control) yeast samples were estimated by transformation of E. coli with the plasmid DNAs recovered from yeast and by assessment of the conversion of the rpsL wild-type phenotype (Sm^S) to its mutant phenotype (Sm^R). After a 40-day space flight, some part of space samples gave mutation frequencies two to three times higher than those of the ground samples. Nucleotide sequence analysis showed no apparent difference in point mutation rates between the space and the ground mutant samples. However, the greater part of the Mir mutant samples were found to have a total or large deletion in the rpsL sequence, suggesting that space radiation containing high-linear energy transfer (LET) might have caused deletion-type mutations.     

Study of the effectiveness of the "Centaur" anti-G suit [correction of suite] during exposure to +Gz accelerations after immersion

This paper describes a study of the effectiveness of the "Centaur" anti-G suit during exposure to +Gz accelerations before and after immersion. Results indicated that the subjects were better able to tolerate the acceleration using the "Centaur" suit than in control studies. Specific cardiovascular responses are presented. This suit has also been used successfully during space missions.     

Effect of long-duration spaceflight on postural control during self-generated perturbations.

This report is the first systematic evaluation of the effects of prolonged weightlessness on the bipedal postural control processes during self-generated perturbations produced by voluntary upper limb movements. Spaceflight impacts humans in a variety of ways, one of which is compromised postflight postural control. We examined the neuromuscular activation characteristics and center of pressure (COP) motion associated with arm movement of eight subjects who experienced long-duration spaceflight (3--6 mo) aboard the Mir space station. Surface electromyography, arm acceleration, and COP motion were collected while astronauts performed rapid unilateral shoulder flexions before and after spaceflight. Subjects generally displayed compromised postural control after flight, as evidenced by modified COP peak-to-peak anterior-posterior and mediolateral excursion, and pathlength relative to preflight values. These changes were associated with disrupted neuromuscular activation characteristics, particularly after the completion of arm acceleration (i.e., when subjects were attempting to maintain upright posture in response to self-generated perturbations). These findings suggest that, although the subjects were able to assemble coordination modes that enabled them to generate rapid arm movements, the subtle control necessary to maintain bipedal equilibrium evident in their preflight performance is compromised after long-duration spaceflight.     

Exercise in Isolation- A Countermeasure for Electrocortical,Mental and Cognitive Impairments

IntroductionMental impairments, including deterioration of mood and cognitive performance, are known to occur during isolation and space missions, but have been insufficiently investigated. Appropriate countermeasures are required, such as exercise, which is known to prevent mood disorders for prolonged space and isolation missions. Based on the interaction of brain activity, mood and cognitive performance, this study aims to investigate the effect of long-term isolation and confinement and the long-term effect of exercise on these parameters.MethodsEight male volunteers were isolated and confined for about eight month during the winter period at the Antarctic Concordia Station. Every six weeks electroencephalographic measurements were recorded under rest conditions, and cognitive tests and a mood questionnaire were executed. Based individual training logs, subjects were afterwards separated into an active (> 2500 arbitrary training units/interval) or inactive (< 2500 arbitrary training units/interval) group.ResultsA long-term effect of exercise was observed for brain activity and mood. Regularly active people showed a decreased brain activity (alpha and beta) in the course of isolation, and steady mood. Inactive people instead first increased and than remained at high brain activity accompanied with a deterioration of mood. No effect of exercise and isolation was found for cognitive performance.ConclusionThe findings point out the positive effect of regularly performed voluntary exercise, supporting subjective mental well-being of long-term isolated people. The choice to be regularly active seems to support mental health, which is not only of interest for future isolation and space missions.     

Gravity independence of seed-to-seed cycling in Brassica rapa

 Growth of higher plants in the microgravity environment of orbital platforms has been problematic. Plants typically developed more slowly in space and often failed at the reproductive phase. Short-duration experiments on the Space Shuttle showed that early stages in the reproductive process could occur normally in microgravity, so we sought a long-duration opportunity to test gravity's role throughout the complete life cycle. During a 122-d opportunity on the Mir space station, full life cycles were completed in microgravity with Brassica rapa L. in a series of three experiments in the Svet greenhouse. Plant material was preserved in space by chemical fixation, freezing, and drying, and then compared to material preserved in the same way during a high-fidelity ground control. At sampling times 13 d after planting, plants on Mir were the same size and had the same number of flower buds as ground control plants. Following hand-pollination of the flowers by the astronaut, siliques formed. In microgravity, siliques ripened basipetally and contained smaller seeds with less than 20% of the cotyledon cells found in the seeds harvested from the ground control. Cytochemical localization of storage reserves in the mature embryos showed that starch was retained in the spaceflight material, whereas protein and lipid were the primary storage reserves in the ground control seeds. While these successful seed-to-seed cycles show that gravity is not absolutely required for any step in the plant life cycle, seed quality in Brassica is compromised by development in microgravity. Received: 3 August 1999 / Accepted: 27 August 1999     

Behavior and reproduction of invertebrate animals during and after a long-term microgravity: space experiments using an Autonomous Biological System (ABS).

Aquatic invertebrate animals such as Amphipods, Gastropods (pond snails), Ostracods and Daphnia (water flea) were placed in water-filled cylindrical vessels together with water plant (hornwort). The vessels were sealed completely and illuminated with a fluorescent lamp to activate the photosynthesis of the plant for providing oxygen within the vessels. Such ecosystem vessels, specially termed as Autonomous Biological System or ABS units, were exposed to microgravity conditions, and the behavior of the animals and their reproduction capacity were studied. Three space experiments were carried out. The first experiment used a Space shuttle only and it was a 10-day flight. The other two space experiments were carried out in the Space station Mir (Shuttle/Mir mission), and the flight units had been kept in microgravity for 4 months. Daphnia produced their offspring during a 10-day Shuttle flight. In the first Mir experiment, no Daphnia were detected when recovered to the ground. However, they were alive in the second Mir experiment. Daphnia were the most fragile species among the invertebrate animals employed in the present experiments. All the animals, i.e., Amphipods, pond snails, Ostracods and Daphnia had survived for 4 months in space, i.e., they had produced their offspring or repeated their life-cycles under microgravity. For the two Mir experiments, in both the flight and ground control ecosystem units, an inverse relationship was noted between the number of Amphipods and pond snails in each unit. Amphipods at 10 hours after the recovery to the ground frequently exhibited a movement of dropping straight-downward to the bottom of the units. Several Amphipods had their legs bent abnormally, which probably resulted from some physiological alterations during their embryonic development under microgravity. From the analysis of the video tape recorded in space, for Ostracods and Daphnia, a half of their population were looping under microgravity. Such looping animals could be observed still at the end of the 4 month stay in space. No looping behavior was noted for Amphipods and pond snails.     

Light Treatment for NASA Shiftworkers

Intense artificial light can phase-shift circadian rhythms and improve performance, sleep, and well-being during shiftwork simulations. In real shiftworkers, however, exposure to sunlight and other time cues may decrease the efficacy of light treatment, and occupational and family responsibilities may make it impractical. With these considerations in mind, we designed and tested light-treatment protocols for NASA personnel who worked on shifted schedules during two Space Shuttle missions. During the prelaunch week, treatment subjects self-administered light of ∼10,000 lux at times of day that phase-delay circadian rhythms. Treatment continued during the missions and for several days afterward. No treatment was administered to subjects in the control group. Treatment subjects reported better sleep, performance, and physical and emotional well-being than control subjects and rated the treatment as highly effective for promoting adjustment to their work schedules. Light treatment is both feasible and beneficial for NASA personnel who must work on shifted schedules during Space Shuttle missions.     

Physiology in microgravity.

Studies of physiology in microgravity are remarkably recent, with almost all the data being obtained in the past 40 years. The first human spaceflight did not take place until 1961. Physiological measurements in connection with the early flights were crude, but, in the past 10 years, an enormous amount of new information has been obtained from experiments on Spacelab. The United States and Soviet/Russian programs have pursued different routes. The US has mainly concentrated on relatively short flights but with highly sophisticated equipment such as is available in Spacelab. In contrast, the Soviet/Russian program concentrated on first the Salyut and then the Mir space stations. These had the advantage of providing information about long-term exposure to microgravity, but the degree of sophistication of the measurements in space was less. It is hoped that the International Space Station will combine the best of both approaches. The most important physiological changes caused by microgravity include bone demineralization, skeletal muscle atrophy, vestibular problems causing space motion sickness, cardiovascular problems resulting in postflight orthostatic intolerance, and reductions in plasma volume and red cell mass. Pulmonary function is greatly altered but apparently not seriously impaired. Space exploration is a new frontier with long-term missions to the moon and Mars not far away. Understanding the physiological changes caused by long-duration microgravity remains a daunting challenge.     

Development of potato minitubers in microgravity

Stem segments of aseptically grown potato (Solanum tuberosum L. cv. Zarevo) were cultivated for 4 weeks under laboratory conditions and were then grown for 8 d on board the "Mir" orbital space station. Timing was such that minitubers initiated and developed during the 8 d on the "Mir". Under space flight and stationary conditions, spherical minitubers were formed with no statistically significant differences in either the frequency of tuber formation or tuber size. These observations are the first to document the formation of vegetative reproductive organs and of well developed amylogenic storage tissue during the microgravity conditions of orbital space flight. In these minitubers, a majority of the starch was stored in parenchyma, with numerous amyloplasts per cell. In space flight tissue, however, grain size of starch was decreased and lamellae within the amyloplasts was locally enlarged. Furthermore, mitochondria of these tissues were characterized by increased matrix density and well developed cristae.     

Review of the Knowledge of Microbial Contamination of the Russian Manned Spacecraft

The 15-year experience of orbital station Mir service demonstrated that specifically modified space vehicle environments allows for the consideration of spaceship habitats as a certain ecological niche of microbial community development and functioning, which was formed from the organisms of different physiological and taxonomical groups. The base unit of the orbital station (OS) Mir was launched on February 20, 1986, and on March 13 the first crew arrived to it. From that moment a unique microbiocenosis started forming in the closed environment of the space station, and vital activity of the microorganisms continued for the next 15 years in a specifically changed environment, in conditions of continuous influence of a set of factors intrinsic to space flight. A total of 234 species of bacteria and fungi were found onboard orbital station Mir, among which microorganisms capable of resident colonization of the environment of space objects as a unique anthropotechnological niche were revealed. In such conditions the evolution of microflora is followed by the rise of medical and technical risks that can affect both sanitary-microbiological conditions of the environment and the safety and reliability characteristics of space equipment. The latter is caused by progressing biological damage to the structural materials. The microbial loading dynamic does not have linearly progressing character, but it is a wavy process of alternation of the microflora activation and stabilization phases; on this background there is a change of the dominating species by quantity and prevalence. The accumulated data is evidence of the necessity of the constant control of the microbial environmental factors to maintain their sanitary and microbiological optimum condition and to prevent the processes of constructional materials biodestruction.