Peptidase-1 expression in some organs of the salamander Pleurodeles waltl submitted to a 12-day space flight.

In Pleurodeles, the peptidase-1 is a sex-linked enzyme encoded by two codominant genes (Pep-1A and Pep-1B) located on the Z and W sex chromosomes. The sexual genotype can be determined by the electrophoretic pattern of the peptidase from erythrocytes. ZAWB genotypic females characterized by 3 electrophoretic bands AA, AB and BB were embarked on Cosmos 2229. The pattern in ovary, muscles and gut issued from the embarked or synchrone females displayed the 3 characteristic bands. In heart and kidney, the bands AA and AB [correction of BB] were revealed, while the band BB appeared very faintly. The specific enzymatic activity in the same organs was compared. Except for the kidney, no statistical significant difference was observed between flight and synchrone samples. This enzyme can be efficiently used as sexual genotypic marker of Pleurodeles experimentally submitted to the effects of space environment.     

Large-size space laboratory for biological orbit experiments.

The study of space factors on living systems has great interest and long-term experiments during orbital flight will be important tool for increasing our knowledge. Realization of such experiments is limited by constraints of modern space stations. A new technology of large-size space laboratory for biological experiments has been developed on the basis of polymerization techniques. Using this technique there are no limits of form and size of laboratory for a space station that will permit long term experiments on Earth orbit with plants and animals in sufficient volume for creation of closed self-regulating ecological systems. The technology is based on experiments of the behavior of polymer materials in simulated free space conditions during the reaction of polymerization. The influences of space vacuum, sharp temperature changes and space plasma generated by galactic rays and Sun irradiation on chemical reaction were evaluated in their impact on liquid organic materials in laboratory conditions. The results of our study shows, that the chemical reaction is sensitive to such space factors. But we believe that the technology of polymerization could be used for the creation of space biological laboratories in Earth orbit in the near future.     

Telescience testbed for biomedical experiments in space morphological and physiological experiments of rat musculoskeletal system.

As the second telescience testbed experiment we were examined sophisticated processes of biomedical experiment, such as an implantation of a transmitter into the hamster's abdominal cavity, non-stressful blood sampling, large amount of blood collection, muscle extirpation and biopsy from the hamsters on February 6-8, 1990. To make clear the differences between successful results obtained by an experienced hand and by a non-experienced one, three operators were selected for three successive experimental days; an engineer who had never experienced any biological experiment, a non-biology student, who experienced on biological experiments, and a veterinary surgeon. Surgical procedures need much experiences on maneuvering and understanding of theory to shorten the elapse time. Especially for a non-experienced hand, graphic instructions were much helpful to understand and to maneuver the procedures. Continuous recordings of ECG from a operator and PIs were of an advantage to grasp an extent of the mental strain, which was compared with their reports requested after end of each experimental day. The mental strain was not related to degrees of scientific achievement, but showed faithfully difficulty of each experimental procedure. Training effects on PIs in successive experimental days were found in their instructions for the operator to let understand the procedures.     

Arthropod model systems for studying complex biological processes in the space environment.

Three arthropod systems are discussed in relation to their complementary and potential use in Space Biology. In a next biosatellite flight, Drosophila melanogaster pre-adapted during several months to different g levels will be flown in an automatic device that separates parental from first and second generations. In the same flight, flies will be exposed to microgravity conditions in an automatic unit in which fly motility can be recorded. In the International Microgravity Laboratory-2, several groups of Drosophila embryos will be grown in Space and the motility of a male fly population will be video-recorded. In the Biopan, an ESA exobiology facility that can be flown attached to the exterior of a Russian biosatellite, Artemia dormant gastrulae will be exposed to the space environment in the exterior of the satellite under a normal atmosphere or in the void. Gastrulae will be separated in hit and non-hit populations. The developmental and aging response of these animals will be studied upon recovery. With these experiments we will be able to establish whether exposure to the space environment influences arthropod development and aging, and elaborate on some of the cellular mechanisms involved which should be tested in future experiments.     

The amphibian egg as a model system for analyzing gravity effects.

Amphibian eggs provide several advantageous features as a model system for analyzing the effects of gravity on single cells. Those features include large size, readily tracked intracellular inclusions, and ease of experimental manipulation. Employing novel gravity orientation as a tool, a substantial data base is being developed. That information is being used to construct a 3-D model of the frog (Xenopus laevis) egg. Internal cytoplasmic organization (rather than surface features) are being emphasized. Several cytoplasmic compartments (domains) have been elucidated, and their behavior in inverted eggs monitored. They have been incorporated into the model, and serve as a point of departure for further inquiry and speculation.     

Crickets in space: morphological, physiological and behavioral alterations induced by space flight and hypergravity.

"Crickets in Space" was a Neurolab experiment by which the balance between genetic programs and the gravitational environment for the development of a gravity sensitive neuronal system was studied. The model character of crickets was justified by their external gravity receptors, identified position-sensitive interneurons (PSI) and gravity-related compensatory head response, and by the specific relation of this behavior to neuronal arousal systems activated by locomotion. These advantages allowed to study the impact of modified gravity on cellular processes in a complex organism. Eggs, 1st, 4th and 6th stage larvae of Acheta domesticus were used. Post-flight experiments revealed a low susceptibility of the behavior to micro- and hypergravity while the physiology of the PSI was significantly affected. Immunocytological investigations revealed a stage-dependent sensitivity of thoracic GABAergic motoneurons to 3 g-conditions concerning their soma sizes but not their topographical arrangement. The morphology of neuromuscular junctions was not affected by 3 g-hypergravity. Peptidergic neurons from cerebral sensorimotor centers revealed no significant modifications by microgravity (micro g). The contrary physiological and behavioral results indicate a facilitation of 1 g-readaptation originating from accessory gravity, proprioceptive and visual sense organs. Absence of anatomical modifications point to an effective time window of micro g or 3 g-expo-sure related to the period of neuronal proliferation. The analysis of basic mechanisms of how animals and man adapt to altered gravitational conditions will profit from a continuation of the project "Crickets in Space".     

Chairman's introduction: mechanisms, models and experiments in space radiation research.

Radiation risk estimate in space is a moral obligation and a scientific challenge requiring the combined efforts of physicists and biologists. This introductory paper presents some thoughts about problems to be solved and the possible directions of research. It stresses the necessity of cooperation across disciplines and the combination of space and ground based investigations.     

Studies on clonogenic hemopoietic cells of vertebrate in space: problems and perspectives.

Hemopoietic tissues were studied in vertebrates launched aboard the Soviet (Russian) biosatellites ("Cosmos-1129, 1514, 1667, 1887 and 2044"; "Bion-10 and 11") between 1980 and 1996. In the bone marrow of rats exposed to spaceflight conditions, a statistically significant decrease in cell number was revealed in the progenitor cell compartment accounting for the compensatory response of granulocyte-macrophage (CFU-gm) and erythrocyte lineages (BFU-e and CFU-e) and in the compartment of multipotent hemopoietic stem cells (CFU-s), which is responsible for the permanent renewal of hemopoietic tissue. The number of stromal fibroblastic progenitors (CFC-f) in the bone marrow of these rats was also reduced. Apparently, changes in the hemopoietic stroma damage the hemopoietic microenvironment and, hence, may be responsible for changes observed in the hemopoietic tissue proper. Attempts were made to develop methods for analyzing morphologically indiscernible clonogenic hemopoietic cells of newts, and studies on the effects of spaceflight factors on these cells were performed. The results showed that the numbers of clonogenic cells in newts of the flight group newts were significantly lower than in control newts. The data obtained are used as the basis for formulating the problems to be studied, drawing up a program for further research on the effects of spaceflight factors on stem and other clonogenic hemopoietic cells, and developing new experimental models for analyzing stem cells, the state of the hemopoietic stroma, etc.     

The pleurodele, an animal model for space biology studies.

Pleurodeles waltl, an Urodele amphibian is proposed as a model for space biology studies. Our laboratory is developing three types of experiments in space using this animal: 1) in vivo fertilization and development ("FERTILE" project); 2) influence of microgravity and space radiation on the organization and preservation of specialized structures in the neurons and muscle cells (in vitro; "CELIMENE" PROJECT); 3) influence of microgravity on tissue regeneration (muscle, bone, epidermis and spinal cord).     

Neuritogenesis: a model for space radiation effects on the central nervous system.

Pivotal to the astronauts' functional integrity and survival during long space flights are the strategies to deal with space radiations. The majority of the cellular studies in this area emphasize simple endpoints such as growth related events which, although useful to understand the nature of primary cell injury, have poor predictive value for extrapolation to more complex tissues such as the central nervous system (CNS). In order to assess the radiation damage on neural cell populations, we developed an in vitro model in which neuronal differentiation, neurite extension, and synaptogenesis occur under controlled conditions. The model exploits chick embryo neural explants to study the effects of radiations on neuritogenesis. In addition, neurobiological problems associated with long-term space flights are discussed.     

Mass balances for a biological life support system simulation model.

Design decisions to aid the development of future space-based biological life support systems (BLSS) can be made with simulation models. Here we develop the biochemical stoichiometry for 1) protein, carbohydrate, fat, fiber, and lignin production in the edible and inedible parts of plants; 2) food consumption and production of organic solids in urine, feces, and wash water by the humans; and 3) operation of the waste processor. Flux values for all components are derived for a steady-state system with wheat as the sole food source. The large-scale dynamics of a materially-closed (BLSS) computer model is described in a companion paper. An extension of this methodology can explore multi-food systems and more complex biochemical dynamics while maintaining whole-system closure as a focus.     

Mathematical model for assessment of radiation risk on long space missions.

A mathematical model is developed which describes the dynamics of radiation-induced mortality in mammalian populations. It relates statistical biometric functions with statistical characteristics and dynamics of an organism's critical system. In the framework of the model the effects of low and very low dose rates of chronic radiation on mice are simulated. Respectively, thrombocytopoietic and granulocytopoietic systems are considered as the critical ones. To calculate the dynamics of these systems, mathematical models are applied, too. In accordance with experimental data, the mortality model reproduces on quantitative level both increased and decreased mortality rates in populations of LAF1 mice, which were chronically exposed, respectively, to low and very low level radiation. All this makes it feasible to use the model as a basis for risk assessments of low level long-term irradiation.     

Simulation experiments of the effect of space environment on bacteriophage and DNA thin films.

The main goal of PUR experiment (phage and uracil response) is to examine and quantify the effect of specific space conditions on nucleic acid models. To achieve this an improved method was elaborated for the preparation of DNA and bacteriophage thin films. The homogeneity of the films was controlled by UV spectroscopy and microscopy. To provide experimental evidence for the hypothesis that interplanetary transfer of the genetic material is possible, phage T7 and isolated T7 DNA thin films have been exposed to selected space conditions: intense UVC radiation (lambda=254 nm) and high vacuum (10(-4) Pa). The effects of DNA hydration, conformation and packing on UV radiation damage were examined. Characteristic changes in the absorption spectrum, in the electrophoretic pattern of DNA and the decrease of the amount of PCR products have been detected indicating the photodamage of isolated and intraphage DNA.     

Biological space experiments for the simulation of Martian conditions: UV radiation and Martian soil analogues.

The survivability of resistant terrestrial microbes, bacterial spores of Bacillus subtilis, was investigated in the BIOPAN facility of the European Space Agency onboard of Russian Earth-orbiting FOTON satellites (BIOPAN I -III missions). The spores were exposed to different subsets of the extreme environmental parameters in space (vacuum, extraterrestrial solar UV, shielding by protecting materials like artificial meteorites). The results of the three space experiments confirmed the deleterious effects of extraterrestrial solar UV radiation which, in contrast to the UV radiation reaching the surface of the Earth, also contains the very energy-rich, short wavelength UVB and UVC radiation. Thin layers of clay, rock or meteorite material were shown to be only successful in UV-shielding, if they are in direct contact with the spores. On Mars the UV radiation climate is similar to that of the early Earth before the development of a protective ozone layer in the atmosphere by the appearance of the first aerobic photosynthetic bacteria. The interference of Martian soil components and the intense and nearly unfiltered Martian solar UV radiation with spores of B. subtilis will be tested with a new BIOPAN experiment, MARSTOX. Different types of Mars soil analogues will be used to determine on one hand their potential toxicity alone or in combination with solar UV (phototoxicity) and on the other hand their UV protection capability. Two sets of samples will be placed under different cut-off filters used to simulate the UV radiation climate of Mars and Earth. After exposure in space the survival of and mutation induction in the spores will be analyzed at the DLR, together with parallel samples from the corresponding ground control experiment performed in the laboratory. This experiment will provide new insights into the principal limits of life and its adaptation to environmental extremes on Earth or other planets which and will also have implications for the potential for the evolution and distribution of life.     

The Rhesus monkey as a model for testing the immunological effects of space flight.

The Rhesus monkey has been proposed as a model for the effects of space flight on immunity. In order to determine the feasibility of the use of the Rhesus monkey as a model, we studied the use of Rhesus monkey cells for immunological procedures that have been shown to be affected by space flight in both rodents and humans. We have shown that both lymph node cells and peripheral blood leukocytes can be stained with monoclonal antibodies to detect the following surface markers: CD4, CD-8, Ia and surface immunoglobulin. Also, the level of Ia antigen expression was increased by treatment of the cells with human interferon-gamma. In addition, cells were induced to produce interferons and interleukins. Isolated neutrophils also demonstrated increased oxidative burst. These data indicate that the Rhesus monkey will be a useful model for space flight studies of immunity.     

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.     

Long-term modulation of Galactic Cosmic Radiation and its model for space exploration.

As the human exploration of space has received new attention in the United States, studies find that exposure to space radiation could adversely impact the mission design. Galactic Cosmic Radiation (GCR), with its very wide range of charges and energies, is particularly important for a mission to Mars, because it imposes a stiff mass penalty for spacecraft shielding. Dose equivalent versus shielding thickness calculations, show a rapid initial drop in exposure with thickness, but an asymptotic behavior at a higher shielding thickness. Uncertainties in the radiobiology are largely unknown. For a fixed radiation risk, this leads to large uncertain ties in shielding thickness for small uncertainties in estimated dose. In this paper we investigate the application of steady-state, spherically-symmetric diffusion-convection theory of solar modulation to individual measurements of differential energy spectra from 1954 to 1989 in order to estimate the diffusion coefficient, kappa (r,t), as a function of time. We have correlated the diffusion coefficient to the Climax neutron monitor rates and show that, if the diffusion coefficient can be separated into independent functions of space and time: kappa (-r,t)=K(t)kappa 0 beta P kappa 1(r), where beta is the particle velocity and P the rigidity, then (i) The time dependent quantity 1/K(t), which is proportional to the deceleration potential, phi(r,t), is linearly related to the Climax neutron monitor counting rate. (ii) The coefficients obtained from hydrogen or helium intensity measurements are the same. (iii) There are different correlation functions for odd and even solar cycles. (iv) The correlation function for the Climax neutron monitor counting rate for given time, t, can be used to estimate mean deceleration parameter phi(t) to within +/- 15% with 90% confidence. We have shown that kappa(r,t) determined from hydrogen and/or helium data, can be used to fit the oxygen and iron differential energy spectra with a root mean square error of about +/- 10%, and essentially independent of the particle charge or energy. We have also examined the ion chamber and 14C measurements which allow the analysis to be extended from the year 1906 to 1990. Using this model we have defined reference GCR spectra at solar minimum and solar maximum. These can be used for space exploration studies and provide a quantitative estimate of the error in dose due to changes in GCR intensities.     

Cell inactivation, repair and mutation induction in bacteria after heavy ion exposure: results from experiments at accelerators and in space.

To understand the mechanisms of accelerated heavy ions on biological matter, the responses of spores of B. subtilis to this structured high LET radiation was investigated applying two different approaches. 1) By the use of the Biostack concept, the inactivation probability as a function of radial distance to single particles' trajectory (i.e. impact parameter) was determined in space experiments as well as at accelerators using low fluences of heavy ions. It was found that spores can survive even a central hit and that the effective range of inactivation extends far beyond impact parameters where inactivation by delta-ray dose would be effective. Concerning the space experiment, the inactivation cross section exceeds those from comparable accelerator experiments by roughly a factor of 20. 2) From fluence effect curves, cross sections for inactivation and mutation induction, and the efficiency of repair processes were determined. They are influenced by the ions characteristics in a complex manner. According to dependence on LET, at least 3 LET ranges can be differentiated: A low LET range (app. < 200 keV/micrometers), where cross sections for inactivation and mutation induction follow a common curve for different ions and where repair processes are effective; an intermediate LET range of the so-called saturation cross section with negligible mutagenic and repair efficiency; and a high LET range (>1000 keV/micrometers) where the biological endpoints are majorly dependent on atomic mass and energy of the ion under consideration.     

Medical prevention of space motion sickness--animal model of therapeutic effect of a new medicine on motion sickness.

Space motion sickness (MS) is one of the most important problems in the field of space medicine. In order to prevent space MS, a new medicine, PMPA, has been prepared by means of synthesizing in our laboratory. The purposes of this study were to set up animal models of PMPA against MS, and to observe its effects on anti-MS, and to prove its function of antagonism to choline. Eight cats, forty rabbits and two hundred and ten rats were selected as animal subjects. The parallel swing stimulus, a method causing the reversal syndromes and tests of anti-choline function were used in our experiments. The results are as follows: (1) The score of MS symptoms in cats with PMPA or scopolamine (SCOP) is significantly lower than that in cats with placebo (p<0.01), while the incidences of efficiency and prevention of PMPA (87.5%, 75%) are higher than those of SCOP (75.0%, 50%) in cats. (2) PMPA of 1.6 mg/kg or 0.8 mg/kg could antagonize the reversal syndromes and repress reversal rotation significantly in rabbits like SCOP in comparison with placebo (p<0.01). (3) PMPA could inhibit tremor evoked by oxotremorine or by nicotine-procaine in rats like SCOP, and play an important role in the antagonism to central M-choline and N-choline receptors. The animal experiments demonstrate that PMPA is an effective medicine against MS with antagonism function to choline.     

Design for a bioreactor with sunlight supply and operations systems for use in the space environment.

An experiment was carried out to determine the characteristics of an operations system that can support fast cultivation of algae at high densities in the weightlessness of space. The experiment was conducted in glass bioreactor tanks, in which light was supplied by radiator rods connected to optical fiber cables. The illumination areas of the tanks were 2600 cm2, 6000 cm2, and 9200 cm2 per liter of solution. The characteristics of O2-CO2 gas exchange, concentration and separation of chlorella in the growth medium, dialysis of ionic salts in the growth medium, etc. were examined. Chloralla ellipsoidea was used in the experiment, yielding the following results: (1) By increasing the ratio of illumination area to volume, growth rates of up to approximately 0.6 g/L h could be obtained in a highly concentrated solution (one that contains 20 g/L or more of algae). (2) The most suitable proportions of carbon dioxide and oxygen gases for growing algae quickly at high concentrations were found to be 10% CO2 and 10% O2 (by volume). (3) There was a high optimum concentration for fast cultivation, and the data obtained resembled the theoretical curve postulated by P. Behrens et al. (4) It was possible to exchange carbon dioxide and oxygen using gas-permeable membrane modules. (5) It was possible to separate the chlorella from the growth medium and recycle the medium.