Effects of altered gravity on plant cell processes: results of recent space and clinostatic experiments.

Space and clinostatic experiments revealed that plant cell structure and metabolism rearrangements depend on taxonomical position and physiological state of objects, growth phase and real or simulated microgravity influence duration. It was shown that clinostat conditions reproduce only a part of microgravity biological effects. It is established that various responses occur in microgravity: 1) rearrangements of cytoplasmic organelles ultrastructure and calcium balance; 2) physical-chemical properties of the plasmalemma are changed; 3) enzymes activity is often enhanced. These events provoke the acceleration of growth and differentiation of cells and their aging as a result; at the same time some responses can be considered as cell adaptation to microgravity.     

Estimation and assessment of Mars contamination.

Since the beginning of the exploration of Mars, more than fourty years ago, thirty-six missions have been launched, including fifty-nine different space systems such as fly-by spacecraft, orbiters, cruise modules, landing or penetrating systems. Taking into account failures at launch, about three missions out of four have been successfully sent toward the Red Planet. The fact today is that Mars orbital environment includes orbiters and perhaps debris, and that its atmosphere and its surface include terrestrial compounds and dormant microorganisms. Coming from the UN Outer Space Treaty [United Nations Treaty on Principles Governing the Activities of States in the Exploration and Use of Outer Space, including the Moon and Other Celestial Bodies (the "Outer Space Treaty") referenced 610 UNTS 205 - resolution 2222(XXI) of December 1966] and according to the COSPAR planetary protection policy recommendations [COSPAR Planetary Protection Policy (20 October 2002), accepted by the Council and Bureau, as moved for adoption by SC F and PPP, prepared by the COSPAR/IAU Workshop on Planetary Protection, 4/02 with updates 10/0, 2002], Mars environment has to be preserved so as not to jeopardize the scientific investigations, and the level of terrestrial material brought on and around Mars theoretically has to comply with this policy. It is useful to evaluate what and how many materials, compounds and microorganisms are on Mars, to list what is in orbit and to identify where all these items are. Considering assumptions about materials, spores and gas location and dispersion on Mars, average contamination levels can be estimated. It is clear now that as long as missions are sent to other extraterrestrial bodies, it is not possible to keep them perfectly clean. Mars is one of the most concerned body, and the large number of missions achieved, on-going and planned now raise the question about its possible contamination, not necessarily from a biological point of view, but with respect to all types of contamination. Answering this question, will help to assess the potential effects of such contamination on scientific results and will address concerns relative to any ethical considerations about the contamination of other planets.     

Biological assessment of Ariane 5 fairing

United Nations Space Treaties [10 and 11] require the preservation of planets and of Earth from contamination. All nations part to these Treaties shall take measures to prevent forward and backward contamination during missions exploring our solar system. As observer for the United Nations Committee on Peaceful Uses of Outer Space, the COSPAR (Committee of Space Research) defines and handles the applicable policy and proposes recommendations to Space Agencies [COSPAR Planetary Protection Panel, Planetary Protection Policy accepted by the COSPAR Council and Bureau, 20 October 2002, amended 24 March 2005. http://www.cosparhq.org/scistr/PPPolicy.htm.]. The goal is to protect celestial bodies from terrestrial biological contamination as well as to protect the Earth environment from an eventual biohazard which may be carried by extraterrestrial samples or by space systems returning to Earth. According to the applicable specifications, including in our case the French requirements [CNES, System Safety. Planetary Protection Requirements. Normative referential CNES RNC-CNES-R-14, CNES Toulouse, ed. 4, 04 October 2002.], the prevention of forward contamination is accomplished by reducing the bioburden on space hardware to acceptable, prescribed levels, including in some instances system sterilization, assembling and integrating the appropriate spacecraft systems in cleanrooms of appropriate biological cleanliness, avoiding or controlling any recontamination risk, and limiting the probability impact of space systems. In order to prepare for future exploration missions [Debus, A., Planetary protection: organization requirements and needs for future planetary exploration missions, ESA conference publication SP-543, pp 103–114, 2003.], and in particular for missions to Mars requiring to control the spacecraft bioburden, a test program has been developed to evaluate the biological contamination under the fairing of the Ariane 5 launcher.     

On the negligible role of manned missions for the geological exploration of Mars: a commentary.

At the 28th Plenary Meeting of the Committee on Space Research (COSPAR) in The Hague, The Netherlands, there was on June 28, 1990, a session of commission MF.1 on Impact of Human Expeditions to Mars, in which, among others, the benefits of manned Mars missions for the geological survey of Mars were discussed. The present commentary does not intend to discuss the pros and cons of manned space flight or of Mars exploration at large, but will reiterate some of the points made in that discussion concerning the justification of manned versus automated Mars exploration in the context of geologic sciences.     

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.     

A consensus approach to planetary protection requirements: recommendations for Mars lander missions.

Over the last several years, the nature of the surface conditions on the planet Mars, our knowledge of the growth capabilities of Earth organisms under extreme conditions, and future opportunities for Mars exploration have been under extensive review in the United States and elsewhere. As part of these examinations, in 1992 the US Space Studies Board made a series of recommendations to NASA on the requirements that should be implemented on future missions that will explore Mars. In particular, significant changes were recommended in the requirements for Mars landers, changes that significantly alleviated the burden of planetary protection implementation for these missions. In this paper we propose a resolution implementing this new set of recommendations, for adoption by COSPAR at its 30th meeting in Hamburg. We also discuss future directions and study areas for planetary protection, in light of changing plans for Mars exploration.     

Implementing planetary protection requirements for sample return missions.

NASA is committed to exploring space while avoiding the biological contamination of other solar system bodies and protecting the Earth against potential harm from materials returned from space. NASA's planetary protection program evaluates missions (with external advice from the US National Research Council and others) and imposes particular constraints on individual missions to achieve these objectives. In 1997 the National Research Council's Space Studies Board published the report, Mars Sample Return: Issues and Recommendations, which reported advice to NASA on Mars sample return missions, complementing their 1992 report, The Biological Contamination of Mars Issues and Recommendations. Meanwhile, NASA has requested a new Space Studies Board study to address sample returns from bodies other than Mars. This study recognizes the variety of worlds that have been opened up to NASA and its partners by small, relatively inexpensive, missions of the Discovery class, as well as the reshaping of our ideas about life in the solar system that have been occasioned by the Galileo spacecraft's discovery that an ocean under the ice on Jupiter's moon Europa might, indeed, exist. This paper will report on NASA's planned implementation of planetary protection provisions based on these recent National Research Council recommendations, and will suggest measures for incorporation in the planetary protection policy of COSPAR.     

Planetary protection issues related to human missions to Mars

In accordance with the United Nations Outer Space Treaties [United Nations, Agreement Governing the Activities of States on the Moon and Other Celestial Bodies, UN doc A/RES/34/68, resolution 38/68 of December 1979], currently maintained and promulgated by the Committee on Space Research [COSPAR Planetary Protection Panel, Planetary Protection Policy accepted by the COSPAR Council and Bureau, 20 October 2002, amended 24 March 2005, http://www.cosparhq.org/scistr/PPPolicy.htm], missions exploring the Solar system must meet planetary protection requirements. Planetary protection aims to protect celestial bodies from terrestrial contamination and to protect the Earth environment from potential biological contamination carried by returned samples or space systems that have been in contact with an extraterrestrial environment. From an exobiology perspective, Mars is one of the major targets, and several missions are currently in operation, in transit, or scheduled for its exploration. Some of them include payloads dedicated to the detection of life or traces of life. The next step, over the coming years, will be to return samples from Mars to Earth, with a view to increasing our knowledge in preparation for the first manned mission that is likely to take place within the next few decades. Robotic missions to Mars shall meet planetary protection specifications, currently well documented, and planetary protection programs are implemented in a very reliable manner given that experience in the field spans some 40 years. With regards to sample return missions, a set of stringent requirements has been approved by COSPAR [COSPAR Planetary Protection Panel, Planetary Protection Policy accepted by the COSPAR Council and Bureau, 20 October 2002, amended 24 March 2005, http://www.cosparhq.org/scistr/PPPolicy.htm], and technical challenges must now be overcome in order to preserve the Earth’s biosphere from any eventual contamination risk. In addition to the human dimension of the mission, sending astronauts to Mars will entail meeting all these constraints. Astronauts present huge sources of contamination for Mars and are also potential carriers of biohazardous material on their return to Earth. If they were to have the misfortune of being contaminated, they themselves would become a biohazard, and, as a consequence, in addition to the technical constraints, human and ethical considerations must also be taken into account.     

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).     

Thin film bioreactors in space.

Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers us an opportunity to learn more about basic biological systems with one important variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would enable us to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.     

Global approach to simulation: a gateway to long-term human presence in space.

The establishment of an autonomous European manned space capability is an objective set up by the ESA Council Meeting at the ministerial level, in 1985/1987. ESA's Long-Term Programme Office (LTPO), charged of the preparation of the programme for a European Manned Space Infrastructure (EMSI), started during 1988 to build up an intellectual framework in the domain of long-duration manned space missions. EMSI scope was eventually extended to embrace Moon/Mars missions and bases. Several exploratory studies on problems related to human factors in long-duration space missions were initiated by LTPO. The work of an ad-hoc group of experts (SIMIS Group) has been focused during 1989/1990 on the planning for simulation of such missions with a broad mandate, covering the physiological, psychological and operational aspects of long-duration exposure to microgravity and isolation/confinement. Preliminary results of SIMIS activities are reported. The HYDREMSI experiment, carried out in a terrestrial, analogous environment for 72 days during 1989, is described as an example of the envisaged simulations.     

ESA microgravity payloads for the 1990s

Analysis of flight opportunities of existing and new microgravity multi-user facilities on Eureca and Spacelab and design studies of new experimental facilities for Columbus are presently in progress.

The materials and fluid sciences research community is likely to be a major user of the permanently manned Space Station/Columbus elements such as the European Attached Pressurised Module (APM) and the Man-Tended Free Flyer (MTFF).

In metallurgy, crystal growth and bioengineering initial research will be performed in the manned laboratory, whereas later on the processing will be automated and executed on unmanned platforms.

At present ESA prepares - in close cooperation with the scientific community - the hardware development of microgravity experimental facilities/laboratories for all Columbus elements through design studies. Preliminary studies which have been carried out to date are the following : Crystallisation Laboratory, Fluid Sciences Laboratory, Containerless Processing Laboratory, Thermophysical Properties Measurement Facility, Metallurgy Laboratory.

In 1998 it is planned to deepen these studies covering laboratories for the APM and for the MTFF. Details on flight opportunities in the pre-Columbus period will also be provided.     

Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space.

Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we intend to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.     

Mammalian cell cultivation in space.

Equipment used in space for the cultivation of mammalian cells does not meet the usual standard of earth bound bioreactors. Thus, the development of a space worthy bioreactor is mandatory for two reasons: First, to investigate the effect on single cells of the space environment in general and microgravity conditions in particular, and second, to provide researchers on long term missions and the Space Station with cell material. However, expertise for this venture is not at hand. A small and simple device for animal cell culture experiments aboard Spacelab (Dynamic Cell Culture System; DCCS) was developed. It provides 2 cell culture chambers, one is operated as a batch system, the other one as a perfusion system. The cell chambers have a volume of 200 microliters. Medium exchange is achieved with an automatic osmotic pump. The system is neither mechanically stirred nor equipped with sensors. Oxygen for cell growth is provided by a gas chamber that is adjacent to the cell chambers. The oxygen gradient produced by the growing cells serves to maintain the oxygen influx by diffusion. Hamster kidney cells growing on microcarriers were used to test the biological performance of the DCCS. On ground tests suggest that this system is feasible.     

Effects of gravity perturbation on developing animal systems

Developing systems provide unique opportunities for analyzing the effects of microgravity on animals. Several unusual types of cells as well as various extraordinary cellular behavior patterns characterize the embryos of most animals. Those features have been exploited as test systems for space flight. The data from previous experiments are reviewed, and considerations for the design of future experiments are presented.     

Progress Update in Space Cell Mechano-biological Coupling

Recent progresses in 2018-2019 from space experiments onboard SJ-10 recoverable satellite and on parabolic flight were summarized, mainly focusing on cell mechano-biological coupling under microgravity. In the meantime, technical pre-research and experimental system design for the biomechanics research platform on China Space Station was carried out and updated.      

Possible mechanisms of plant cell wall changes at microgravity.

Space and clinostatic experiments revealed that changes of plant cell wall structure and its function depend on type of tissue and duration of influence. It was shown that clinostat conditions reproduce the part of weightlessness biological effects. It is established that various responses of wall structural-metabolic organization occur at microgravity: changes of cell walls ultrastructure and organelles structure; decrease of synthesis of primary plant cell wall; rearrangements of polysaccharides content. It is shown that mechanisms of plant cell wall changes at microgravity are connected with decrease of cellulose crystallization, activation of pectolytic enzymes and rearrangement of calcium balance of apoplast and cytoplasm.     

Reports to COSPAR from the International Lunar Exploration Working Group (ILEWG)

In accordance with its charter, the International Lunar Exploration Working Group (ILEWG) reports to COSPAR, and a summary was given at the Beijing COSPAR 2006 Assembly on ILEWG activities conducted since the previous COSPAR 2004 assembly held in Paris. This included reports from the 6th and 7th ILEWG International Conference on Exploration and Utilization of the Moon, held respectively in Udaipur, India on 22–26 November 2004 (ICEUM6) and in Toronto, Canada on 18–23 September 2005 (ICEUM7). We give in this issue of Advances in Space Research the “lunar declarations” from these ICEUM conferences, as well as for the ICEUM8 conference held in Beijing immediately after the 2006 COSPAR Assembly. One year after the COSPAR Beijing assembly, the 9th ILEWG International Conference on Exploration and Utilization of the Moon (ICEUM9), was held in Sorrento, Italy on 18–23 September 2007. We report also in this issue the “Sorrento Lunar Declaration” in advance of the ILEWG formal report to be given at the COSPAR Assembly to be held in Montreal, Canada in July 2008.