Short-term responses of gravitaxis to altered gravity in Paramecium.

Negative gravitaxis of Paramecium almost disappeared in solutions having specific gravity about the same as that of the organisms (1.04). The taxis turned to positive in solutions of specific gravity 1.08. Using a drop shaft at the Japan Microgravity Center, Hokkaido (JAMIC) we examined how swimming behaviour in these media was modified by changing gravitational conditions before, during and after free-fall. Tracks of swimming cells recorded on videotape indicate that the swimming cells continued upward and downward shift depending on the specific gravity of the external medium under 1-g conditions and these vertical displacements disappeared immediately after the moment of launch. The effectiveness of changing gravity to induce displacement of the cells seems to depend on the orientation of the cells to gravity. These results suggest a corelation between vertical displacement of the cell through the medium and a gravitactic mechanism in Paramecium.     

Effects of gravity and cosmic rays on cell proliferation kinetics in Paramecium tetraurelia.

Space flights resulted in a stimulating effect on kinetics of proliferation in Paramecium tetraurelia. Additional experiments were performed in order to determine the origin of this phenomena. Paramecia were cultivated in balloon flights or in a slow clinostat, or were exposed to different levels of hypergravity. The results suggest that changes in cell proliferation rate are related to cosmic rays and to a direct effect of microgravity.     

Biological role of gravity: hypotheses and results of experiments on "Cosmos" biosatellites.

In order to reveal the biological significance of gravity, microgravity effects have been studied at the cellular, organism and population levels. The following questions arise. Do any gravity-dependent processes exist in a cell? Is cell adaptation to weightlessness possible; if so, what role may cytoskeleton, the genetic apparatus play in it? What are the consequences of the lack of convection in weightlessness for the performance of morphogenesis? Do the integral characteristics of living beings change in weightlessness? Is there any change in "biological capacity" of space, its resistance to expansion of life? What are the direction and intensity of microgravity action as a factor of natural selection, the driving force of evolution? These problems are discussed from a theoretical point of view, and in the light of results obtained in experiments from aboard biosatellites "Cosmos".     

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.     

Intramuscular calcium movements: experiments from the Soviet biosatellite Biocosmos.

Experiments have been performed in skeletal muscle fibres from the lateral head of gastrocnemius muscle of female rats. Changes in intramuscular calcium movements due to microgravity conditions have been tested by tension measurements in chemically skinned muscle fibres. Our results show that microgravity induces i) a decrease in maximal muscle strength developed by contractile proteins ii) a decrease of intensity and rate of both Ca release and Ca uptake by the sarcoplasmic reticulum.     

Radiation effects in nematodes: results from IML-1 experiments.

The nematode Caenorhabditis elegans was exposed to natural space radiation using the ESA Biorack facility aboard Spacelab on International Microgravity Laboratory 1, STS-42. For the major experimental objective dormant animals were suspended in buffer or on agar or immobilized next to CR-39 plastic nuclear track detectors to correlate fluence of HZE particles with genetic events. This configuration was used to isolate mutations in a set of 350 essential genes as well as in the unc-22 structural gene. From flight samples 13 mutants in the unc-22 gene were isolated along with 53 lethal mutations from autosomal regions balanced by a translocation eT1(III;V). Preliminary analysis suggests that mutants from worms correlated with specific cosmic ray tracks may have a higher proportion of rearrangements than those isolated from tube cultures on a randomly sampled basis. Right sample mutation rate was approximately 8-fold higher than ground controls which exhibited laboratory spontaneous frequencies.     

Super-helix model: a physiological model for gravitaxis of Paramecium.

A new model explaining the gravitactic behavior of Paramecium is derived on the basis of its mechanism of gravity sensing. Paramecium is know to have depolarizing mechanoreceptor ion channels in the anterior and hyperpolarizing channels in the posterior of the cell. This arrangement may lead to bidirectional changes of the membrane potential due to the selective deformation of the anterior and posterior cell membrane responding to the orientation of the cell with respect to the gravity vector; i.e., negative- and positive-going shifts of the potential due to the upward and downward orientation, respectively. The orientation dependent changes in membrane potential, in combination with the close coupling between the membrane potential and ciliary locomotor activity, may allow the changes in swimming direction along the otherwise simple helical swimming path in the following manner: an upward shift of the axis of helical swimming occurs by decreasing the pitch angle due to channel-dependent hyperpolarization in upward-orienting cells, and an upward shift of the swimming helix occurs by increasing the cell's pitch angle due to depolarization in downward-orienting cells. Computer simulation of the model demonstrated that the cell can swim upward along the "super-helical" trajectory consisting of a small helix winding helically along an axis parallel to the gravity vector.     

Lessons from laboratory experiments on reconnection

Magnetic reconnection has been studied in a laboratory experiment designed to model the basic two-dimensional neutral sheet configuration. However, the focus has been put on the inner region of the neutral sheet where the ions are effectively unmagnetized and MHD concepts are violated. In this parameter regime driven reconnection is governed by the fast dynamics of electrons. In true neutral sheets (B_z ⋍ 0) the current is carried by electrons. Thin current sheets (Δz ≳ c/ω_pe) rapidly form multiple X and 0 points due to the onset of the collisionless electron tearing mode. Magnetic energy is transported along the separator at the speed of whistler waves rather than Alfvén waves. Due to space charge separation the reconnection electric field E_y is, in general, not constant along the separator but localized near boundaries, nonuniformities in density and magnetic fields which limit the current I_y. This leads to localized particle acceleration, formation of anisotropic velocity distributions and instabilities. Reconnection and energization can be spatially separated which shows the importance of investigating both the global current system as well as critical local plasma properties. Experiments of current sheet disruptions are performed which demonstrate the processes of magnetic energy storage, transport, conversion and dissipation. Double layers and shock waves can be produced by current disruptions. The laboratory experiments show new dynamic features of reconnection processes not considered in MHD models yet relevant to narrow current sheets or the center of thick sheets.     

Living plant systems: How robust are they in the absence of gravity?

The following hierarchical levels can be recognised in plant systems: cells, organs, organisms and gamodemes (interbreeding members of a community). Each level in this ‘living hierarchy’ is both defined and supported by a similar set of sub-systems. Within this framework of plant organization, two complementary questions are relevant for interpreting plant-oriented space experiments: 1) What role, if any, does gravity play in enabling the development of each organizational level? and 2) Does abnormal development in an altered gravity environment indicate sub-system inefficiency? Although a few representatives of the various organizational levels in plant systems have already been the subject of microgravity experiments in space laboratories—from cells in culture to gamodemes, the latter being found in some Closed Environment Life Support Systems—it would be of interest to investigate additional systems with respect to their response to microgravity. Recognition of the sub-systems at each level might be relevant not only for a more complete understanding of plant development but also for the successful cultivation and propagation of plants during long-term space flights and the establishment of plants in extra-terrestrial environments.     

Effects of gravity on early development.

The development of embryonic and larval stages of the South African Toad Xenopus laevis D, was investigated in hyper-g up to 5 g (centrifuge), in simulated 0 g (fast-rotating clinostat), in alternating low g, hyper-g (parabolic flights) and in microgravity (Spacelab missions D1, D-2). The selected developmental stages are assumed to be very sensitive to environmental stimuli. The results showed that the developmental reaction processes run normal also in environments different to 1 g and that aberrations in behavior and morphology normalize after return to 1 g. Development, differentiation, and morphology of the gravity perceiving parts of the vestibular system (macula-organs) had not been affected by exposure to different g-levels.     

The effect of gravity on plant germination.

An axis clinostat was constructed to create micro and negative gravity also a rotated flat disk was constructed with different rotation rates to give increased gravity, by centrifugal force up to 48 g. Rice seeds were grown on agar in tubes at the constant air temperature of 20 degrees C under an average light condition of 110 micromol/m2/sec(PPF). Humidity was not controlled but was maintained above 90%. Since the tube containers were not large enough for long cultivation, shoot and root growth were observed every 12 hours until the sixth day from seeding. The lengths of shoots and roots for each individual plant were measured on the last day. The stem lengths were increased by microgravity but the root lengths were not. Under the negative gravity, negative orthogeotropism and under microgravity, diageotropism was observed. No significant effect of increased gravity was observed on shoot and root growth.     

Weightlessness acts on human breast cancer cell line MCF-7.

Because cells are sensitive to mechanical forces, weightlessness might act on stress-dependent cell changes. Human breast cancer cells MCF-7, flown in space in a Photon capsule, were fixed after 1.5, 22 and 48 h in orbit. Cells subjected to weightlessness were compared to 1 g in-flight and ground controls. Post-flight, fluorescent labeling was performed to visualize cell proliferation (Ki-67), three cytoskeleton components and chromatin structure. Confocal microscopy and image analysis were used to quantify cycling cells and mitosis, modifications of the cytokeratin network and chromatin structure. Several main phenomena were observed in weightlessness: The perinuclear cytokeratin network and chromatin structure were looser; More cells were cycling and mitosis was prolonged. Finally, cell proliferation was reduced as a consequence of a cell-cycle blockade; Microtubules were altered in many cells. The results reported in the first point are in agreement with basic predictions of cellular tensegrity. The prolongation of mitosis can be explained by an alteration of microtubules. We discuss here the different mechanisms involved in weightlessness alteration of microtubules: i) alteration of their self-organization by reaction-diffusion processes, and a mathematical model is proposed, ii) activation or deactivation of microtubules stabilizing proteins, acting on both microtubule and microfilament networks in cell cortex.     

Possible actions of gravity on the cellular machinery.

Since the first flight of the ESA Biorack on the German Spacelab Mission D1 in 1985 evidence has been obtained that biological cells and small unicellular organisms function differently under conditions of microgravity. However, there is still lack of scientific proof that these effects are caused by a direct influence on the cells in the weightlessness condition. The question how normal gravity may play a role in cellular activity is being addressed and the results show that gravity may provide important signals during certain state transitions in the cell. These would be gravity-sensitive windows in the biological process. Also, by amplification mechanisms inside the cell, the cell may assume a state that is typical for normal gravity conditions and would change in microgravity. Experimental tools are discussed that would provide the conditions to obtain evidence for direct action of gravity and for the possible existence of gravity-sensitive windows.     

An improved altimeter-derived gravity anomaly from shipborne gravity based on the mean sea surface height constraint factors method

Coastal marine gravity modeling faces challenges due to the degradation of the quality and poor coverage of altimeter data in coastal regions. The effective fusion of shipborne gravity data and altimeter-derived marine gravity data can make shipborne gravity data more useful for the accurate estimation of altimeter-derived coastal marine gravity. A mean sea surface height constraint factor (MSSHCF) method based on the ordinary kriging method and the remove-restore technique is proposed to fuse altimeter-derived gravity model with shipborne gravity data. In this method, all data are standardized during the interpolation process to reduce the error and mean sea surface as a vertical variable is added to the semi-variance function in ordinary kriging to obtain the residual shipborne gravity as corrected data source. The coastal marine gravity models V2.1 and V3.1 which fused altimeter-derived gravity data with shipborne gravity data and V1.1 without shipborne gravity data at a spatial resolution of 1′×1′ can be obtained. Validation experiments show that the accuracy of the gravity model V3.1 obtained by the MSSHCF method more closely agrees with the validated gravity model DTU17 and SS V31 than the model V2.1 obtained by the ordinary kriging interpolation method and the V1.1 model. Our results were validated against shipborne gravity data; the accuracy of model V3.1 was 4.95 % higher than the model V1.1 in South China Sea area A and 2.48 % higher in South China Sea area B. Meanwhile, the accuracy of model V3.1 was 2.07 % higher than model V2.1 in South China Sea area A and 2.42 % higher in South China Sea area B. The effects of distance from the coast and sea depth on the marine gravity model were also evaluated. The results show that the gravity model V3.1 has higher accuracy with the change in ocean distance and depth than the V2.1 and V1.1 gravity models. Thus, our study shows that the MSSHCF method effectively refines coastal altimeter-derived gravity using shipborne gravity data.     

Influence of gravity on the circadian timing system.

The circadian timing system (CTS) is responsible for daily temporal coordination of physiological and behavioral functions both internally and with the external environment. Experiments in altered gravitational environments have revealed changes in circadian rhythms of species ranging from fungi to primates. The altered gravitational environments examined included both the microgravity environment of spaceflight and hyperdynamic environments produced by centrifugation. Acute exposure to altered gravitational environments changed homeostatic parameters such as body temperature. These changes were time of day dependent. Exposure to gravitational alterations of relatively short duration produced changes in both the homeostatic level and the amplitude of circadian rhythms. Chronic exposure to a non-earth level of gravity resulted in changes in the period of the expressed rhythms as well as in the phase relationships between the rhythms and between the rhythms and the external environment. In addition, alterations in gravity appeared to act as a time cue for the CTS. Altered gravity also affected the sensitivity of the pacemaker to other aspects of the environment (i.e., light) and to shifts of time cues. Taken together, these studies lead to the conclusion that the CTS is indeed sensitive to gravity and its alterations. This finding has implications for both basic biology and space medicine.     

Cellular responses to gravity: extracellular, intracellular and in-between.

Our understanding of gravitational effects (inertial effects in the vicinity of 1 x g) on cells has matured to a stage at which it is possible to define, on the basis of experimental evidence, extracellular effects on small cells and intracellular effects on eukaryotic gravisensing cells. Yet undetermined is the nature of response, if any, of those classes of cells that are not governed solely by extracellular physical events (as are prokaryotes) and are devoid of obvious mechanical devices for sensing inertial forces (such as those possessed by certain plant cells and sensory cells of animals). This "in-between" class of cells needs to be understood on the basis of the combination of intracellular and extracellular gravity-dependent processes that govern experimentally-measurable variables that are relevant to the cell's responses to modified inertial forces. The forces that certain cell types generate or respond to are therefore compared to those imposed by approximately 1 x g in the context of cytoskeletal action and symmetry-breaking pathways.     

Synaptic plasticity and gravity: ultrastructural, biochemical and physico-chemical fundamentals.

On the basis of quantitative disturbances of the swimming behaviour of aquatic vertebrates ("loop-swimming" in fish and frog larvae) following long-term hyper-g-exposure the question was raised whether or not and to what extent changes in the gravitational vector might influence the CNS at the cellular level. Therefore, by means of histological, histochemical and biochemical analyses the effect of 2-4 x g for 9 days on the gross morphology of the fish brain, and on different neuronal enzymes was investigated. In order to enable a more precise analysis in future-microgravity-experiments of any gravity-related effects on the neuronal synapses within the gravity-perceptive integration centers differentiated electron-microscopical and electronspectroscopical techniques have been developed to accomplish an ultrastructural localization of calcium, a high-affinity Ca2(+)-ATPase, creatine kinase and cytochrome oxidase. In hyper-g animals vs. 1-g controls, a reduction of total brain volume (15%), a decrease in creatine kinase activity (20%), a local increase in cytochrome oxidase activity, but no differences in Ca2+/Mg(2+)-ATPase activities were observed. Ultrastructural peculiarities of synaptic contact formation in gravity-related integration centers (Nucleus magnocellularis) were found. These results are discussed on the basis of a direct effect of hyper-gravity not only on the gravity-sensitive neuronal integration centers but possibly also on the physico-chemical properties of the lipid bilayer of neuronal membranes in general.     

Effects of altered gravity on the swimming behaviour of fish.

Humans taking part in parabolic aircraft flights (PAFs) may suffer from space motion sickness-phenomena (SMS, a kinetosis). It has been argued that SMS during PAFs might not be based on microgravity alone but rather on changing accelerations from 0 g to 2 g. We test here the hypothesis that PAF-induced kinetosis is based on asymmetric statoliths (i.e., differently weighed statoliths on the right and the left side of the head), with asymmetric inputs to the brain being disclosed at microgravity. Since fish frequently reveal kinetotic behaviour during PAFs (especially so-called spinning movements and looping responses), we investigated (1) whether or not kinetotically swimming fish at microgravity would have a pronounced inner ear otolith asymmetry and (2) whether or not slow translational and continuously changing linear (vertical) acceleration on ground induced kinetosis. These latter accelerations were applied using a specially developed parabel-animal-container (PAC) to stimulate the cupular organs. The results suggest that the fish tested on ground can counter changing accelerations successfully without revealing kinetotic swimming patterns. Kinetosis could only be induced by PAFs. This finding suggests that it is indeed microgravity rather than changing accelerations, which induces kinetosis. Moreover, we demonstrate that fish swimming kinetotically during PAFs correlates with a higher otolith asymmetry in comparison to normally behaving animals in PAFs.     

Longevity in space; experiment on the life span of Paramecium cell clone in space.

Life span is the most interesting and also the most important biologically relevant time to be investigated on the space station. As a model experiment, we proposed an investigation to assess the life span of clone generation of the ciliate Paramecium. In space, clone generation will be artificially started by conjugation or autogamy, and the life span of the cell populations in different gravitational fields (microgravity and onboard 1 x g control) will be precisely assessed in terms of fission age as compared with the clock time. In order to perform the space experiment including long-lasting culture and continuous measurement of cell division, we tested the methods of cell culture and of cell-density measurement, which will be available in closed environments under microgravity. The basic design of experimental hardware and a preliminary result of the cultivation procedure are described.