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Gravitaxis in the flagellate Euglena gracilis--results from NiZeMi, clinostat and sounding rocket flights.

Many motile microorganisms including flagellates such as the green Euglena gracilis move up and down within the water column and use a number of external clues for their orientation, the most important of which may be light and gravity. The cells use positive phototaxis and negative gravitaxis to move closer to the surface of the water column which for energetic reasons is vital for their survival. However, most phytoplankton organisms cannot tolerate the bright irradiance of unfiltered solar radiation at the surface which also bleaches the photosynthetic pigments, disables the photosynthetic apparatus and impairs phototaxis, gravitaxis and motility in Euglena. Thus, it is not surprising that at higher irradiances negative phototaxis operates antagonistically to the responses described above to guide the cells into deeper water where they are protected from excessive radiation. Phototaxis and gravitaxis are not independent from one another: in a vertically positioned cuvette negative gravitaxis can be "titrated" by light impinging from above and is compensated at about 30 W m-2. While the photoreceptor for phototaxis has been identified in Euglena gracilis biochemically and spectroscopically, the gravireceptor is not yet known. Young cultures of Euglena gracilis show a positive gravitaxis, the ecological signficance of which is not yet understood while older cultures show negative gravitaxis. One hypothesis concerning the nature of graviperception is based on a passive physical process such as an asymmetric distribution of the mass within the cell. However, the observation that short term UV irradiation decreases the precision of negative gravitaxis rather indicates the involvement of an active physiological gravireceptor. Furthermore, some heavy metal ions have been found to change the direction of movement from positive to negative gravitaxis in young cells.

Animals↗

Behavior of free-swimming cells under various accelerations.

The different steps of the gravity signal-transduction chain on the cellular level are not identified. In our experiments performed up to now we mainly stressed our attention on the last step, the response of the cells. Swimming behavior is a suitable indicator for the physiological status of a Paramecium cell. Depending on membrane potential and/or concentrations of Ca++, cGMP and cAMP the beating direction and the beating velocity of the cilia are influenced in a characteristical way leading to a changed swimming activity of the cell. The behavior of Paramecium is influenced by various stimuli from their environment. Previous studies have demonstrated that under controlled conditions Paramecium shows a clear gravity-dependent behavior resulting in negative gravitaxis and gravikinesis (speed regulation in dependence of gravity). By changing the orienting stimulus (gravity) we expected changes of the swimming behavior. Additional experiments were performed using pawn mutant d4-500r. Due to defective Ca(2+)-channels the membrane of this mutant cannot depolarize. As a consequence d4-500r cannot perform phobic responses and swim backwards. Comparative experiments are also performed with the ciliate Loxodes striatus. In contrast to Paramecium this ciliate possesses statocyst-like organelles--the Müller Organelles.

Animals↗

Parabolic flight experiments on physiological data acquisition and processing technologies using small jet aircraft (MU300).

The parabolic aircraft flight provides a short low gravity environment for approximately 20 seconds, which may not be sufficient for a research on the physiological phenomenon induced by actual weightlessness in space. However, the method is still useful to reveal essential and characteristic feature of physiological signs, and is available for testing hardware and also training of crew member during altered gravity. This paper reports the summary of parabolic flight experiments recently conducted as a NASDA program (1990-1992). The program is providing opportunities in low gravity research with small jet aircraft for researchers and agencies. The flight experiments in the life science area have been conducted mostly focused on a physiological changes and basic methodology which may be effective under the altered gravity condition. In this study, the following research team, NASDA, Research Institute of Environmental Medicine, Nagoya University, Toyohashi University of Technology, Tokyo Metropolitan Hospital, Torey Research Center and JSUP were involved and coordinated for the research.

Aircraft↗

Whole body movements in altered G environments.

Davies and Rennie determined that the instantaneous peak and mean power transferred to the center-of-mass of a subject during jumping could be calculated from analyzing the ground reaction force. Since Davies and Rennie's report, vertical jumping has been used to measure the peak and mean power transferred to the center-of-mass and to assess human performance. It has also been suggested that the external load a subject experiences effects the power production. However, it is difficult to determine if the difference in power production as defined in these studies is due to the external loading, or is an effect of changing the dynamics of the movement. In this paper, we describe a study which allows us to more clearly define the power developed during jumping under different weights, and the relationship between the power developed and jump height.

Adult↗

The study on the mechanism of G intolerance of rabbits after simulated weightlessness.

The direct consequence of cardiovascular adaptation to weightlessness (WL) is the decrease of G tolerance. In studying the mechanism of G intolerance after WL, respiration, heart rate, electrocardiogram, temporal arterial flow, loss of vision were usually used as the indices for evaluation of G tolerance. However the changes of microcirculation and blood rheological indices were seldom observed. Considering that the changes of status of blood circulation after WL may be one of the important factors causing decrease of G tolerance, the purpose of this paper is to observe the changes of microcirculation, blood rheological and the structure and circulatory status of four organs in rabbits during -4Gx after exposure to simulated weightlessness (SWL), in order to understand the cause of G intolerance after WL.

Animals↗

Application of multivariate statistical analysis to estimate +Gz tolerance based on the changes of hemodynamic parameters during lower body negative pressure (LBNP).

The application of lower body negative pressure (LBNP) is very useful method for simulation of +Gz stress and for evaluation of orthostatic reaction. The different physiological changes that occur during LBNP test and +Gz acceleration test are similar. Lategola and Trent found that supine LBNP exposure at the level of -50 mmHg may be equivalent to +2Gz in producing the changes of heart rate (HR). Polese and coworkers compared hemodynamic changes occurring during upright and supine LBNP at the levels to -70 mmHg with identical measurements made during accelerations to +2Gz, +3Gz, and +4Gz in the same subjects. They noted for example that HR changes during upright LBNP exceeded HR supine levels. Peak values of HR during +3Gz and +4Gz significantly exceeded HR levels during both kinds of LBNP, but HR values at +2Gz were equivalent to those at -40 mmHg of upright and -70 mmHg of supine LBNP. So, the present study was undertaken to evaluate adaptating responses to LBNP stimulus at the level of -60 mmHg, regulatory mechanisms of the circulatory system (central and peripheral) and to look for the possibility of +Gz tolerance prediction based on the changes of some hemodynamic parameters during LBNP.

Adaptation, Physiological↗

Relationship between atrial natriuretic peptide (ANP), renin (PRA), aldosterone (PAC), hemodynamic responses to lower body negative pressure (LBNP) and +Gz tolerance.

The redistribution of a certain thoracic blood volume to the lower parts of the body and decrease of the venous return of blood to the heart during lower body negative pressure leads to the central hypovolemia and the deactivation of cardiopulmonary and arterial baroreceptors. Many compensatory mechanisms are involved during central hypovolemia, which is also reflected by the changes in the secretion of different vasoactive hormones. Due to this fact the LBNP stimulus is widely used for the investigation of regulatory (compensatory) mechanisms in cardiovascular system providing deeper understanding of orthostatic reaction. Recently several papers were published on application of this experimental model for +Gz acceleration tolerance assessment. The purpose of this study was evaluate the possible dependence between the changes of ANP secretion, renin-angiotensin-aldosterone system activity, the changes of some hemodynamic parameters during the model of gravitational stress i.e. LBNP exposure and +Gz acceleration tolerance.

Adaptation, Physiological↗

Acute exposure to 2G phase shifts the rat circadian timing system.

The circadian timing system (CTS) provides internal and external temporal coordination of an animal's physiology and behavior. In mammals, the generation and coordination of these circadian rhythms is controlled by a neural pacemaker, the suprachiasmatic nucleus (SCN), located within the hypothalamus. The pacemaker is synchronized to the 24 hour day by time cues (zeitgebers) such as the light/dark cycle. When an animal is exposed to an environment without time cues, the circadian rhythms maintain internal temporal coordination but exhibit a "free-running" condition in which the period length is determined by the internal pacemaker. Maintenance of internal and external temporal coordination are critical for normal physiological and psychological function in human and non-human primates. Exposure to altered gravitational environments has been shown to affect the amplitude, mean, and timing of circadian rhythms in species ranging from unicellular organisms to man. However, it has not been determined whether altered gravitational fields have a direct effect on the neural pacemaker, or affect peripheral physiological systems that express these circadian parameters. In previous studies, the ability of a stimulus to phase shift circadian rhythms was used to determine whether a stimulus has a direct effect on the neural pacemaker. The present experiment was performed in order to determine whether acute exposure to a hyperdynamic field could phase shift circadian rhythms.

Animals↗

Acceleration of fusion in mouse palates by in vitro exposure to excess G.

Palatal shelves from 13- and 14-day mouse embryos were excised and cultured in contiguous pairs. Experimental cultures were exposed to 2.6 G in a culture centrifuge; controls were in the same incubator. After 24 hours, palates were prepared for light or electron microscopy. Scoring of paraffin sections according to the stage of fusion seen in the medial epithelial edges (MEE) showed that palates exposed to excess G were in more advanced stages of fusion than were controls. Ultrastructurally, control MEE had tightly apposed cell membranes and numerous desmosomes; in centrifuged MEE, desmosomes had been removed and there was much intercellular space. Nuclear membranes were intact in control MEE, but showed marked deterioration in MEE of centrifuged palates. Few lysosomes and no necrosis were seen in control MEE; centrifuged MEE had numerous lysosomes as well as necrotic cells. Basal lamina were intact in controls, but interrupted in centrifuged palates. The results confirm the hypothesis that gravitational increases speed up the differentiative process.

Animals↗

The primate circadian timekeeping system in a hyperdynamic environment.

This study was designed to examine the possible effects of hyperdynamic fields (+G) on the circadian timekeeping system of the squirrel monkey. Eight monkeys were sequentially exposed to 1.0, 1.5, 2.0 and 1.0 G. During the two 1 G phases and the 2.0 G phase, the animals were exposed to constant light (LL) in addition to the 24-hr light-dark cycle (LD 12:12) present during the rest of the experiment. All animals were housed simultaneously on an 18 ft diameter centrifuge. The animals were allowed food and water ad lib, the intake of which was monitored electronically every 30 minutes. In all three acceleration fields, the animals demonstrated prominent circadian rhythms in both feeding and drinking. In the LD cycle, both rhythms were entrained with 24 hr periods. A phase delay occurred between the rhythms and the LD synchronizer in the +G environment. Moreover, when the animals were exposed to LL at 2.0 G, the mean free-running period was longer than when the animals were at 1 G either before or after the hyperdynamic exposure. In summary, this study demonstrates that the circadian timekeeping system of these primates is functional in the hyperdynamic environment, however, some of its components appear to be regulated at different homeostatic levels.

Activity Cycles↗

Gravitational physiology of human immune cells: a review of in vivo, ex vivo and in vitro studies.

The study of the function of immune cells in microgravity has been studied for more than 20 years in several laboratories. It is clear today that the immune system is depressed in more than 50% of the astronauts during and after space flight and that the activation of T lymphocytes by mitogens in vitro changes dramatically. This article gives an overview of the gravitational studies conducted by our laboratory in Spacelab, in MIR station, in sounding rockets and on the ground in the clinostat and the centrifuge. Three experimental approaches are followed in our work: (i) Ex vivo studies are performed with blood samples drawn from astronauts; (ii) in vivo studies are based on the application of seven antigens to the skin of the astronauts; (iii) in vitro studies are carried out with immune cells purified from the blood of healthy donors (not astronauts). The data from our in vivo and ex vivo studies are in agreement with those of other laboratories and show that the immunological function is depressed in the majority of astronauts as a consequence of the stress of space flight rather than by a direct influence of gravity on the cell. Immune depression may become a critical hazard on long duration flights on space stations or to other planets. In vitro experiments show that cultures of free-floating lymphocytes and monocytes undergo a dramatic depression of activation by the mitogen concanavalin A, while activation is more than doubled when the cells are attached to microcarrier beads. Such effects may be attributed to both direct and indirect effects of gravitational unloading on basic biological mechanisms of the cell. While the in vitro data are very important to clarify certain aspects of the biological mechanism of T cells activation, they are not descriptive of the changes of the immunological function of the astronauts.

Animals↗

Expression of c-fos, c-jun and HSP70 mRNA in rat brain following high acceleration stress.

Rats exposed to high +Gz forces in a small animal centrifuge (SAC) exhibit loss of neuronal function (isoelectric EEG), termed G-induced loss of consciousness (G-LOC). This phenomenon is presumably due to a reduction in cerebral blood flow (CBF) or ischemia. Ischemia induces various metabolic and physiologic changes including expression of immediate early genes (IEGs) in the brain. Expression of IEGs have been suggested to be reliable markers for neuronal response to external stimuli or stress. In the present study expression of IEGs c-fos, c-jun and stress response gene HSP70 were measured in the brains of rats subjected to six 30 s exposures of +22.5Gz in a small animal centrifuge. The level of c-fos, HSP70 and beta-actin mRNA were measured by both Northern blot and RT-PCR. Expression of c-jun was measured only by RT-PCR. Expression of c-fos and c-jun was significantly stimulated at 0.5, 15, 30 and 60 min post-centrifugation. The level of HSP70 mRNA was significantly higher only at 60 and 180 min post-centrifugation. Measurement of metabolities showed a significant increase in lactate and a decrease in Cr-P level at 30 s and 15 min post-centrifugation, respectively. Lactate, but not Cr-P and ATP levels were restored to control levels by 60 min post-centrifugation. It is concluded that the transient expression of c-fos, c-jun and HSP70 mRNA is stimulated by repeated ischemic/reperfusion episodes induced by high acceleration stress.

Acceleration↗

[Cerebral cortical blood flow during parabolic maneuvers].

Cerebral cortical blood flow (CBF) was measured in 6 rabbits during parabolic flight maneuvers (20 s of weightlessness). During the first seconds of weightlessness, CBF increased up to 100% when compared with CBF at 1 G. Since this transient increase (5 s) was not modified when propranolol or pentobarbital was given to the animals, one can hypothesize that it was not related to stress.

Animals↗

Predicting skeletal adaptation in altered gravity environments.

It is generally agreed that the single factor that most limits human survivability in non-Earth environments is the phenomenon of bone demineralization and the medical problems induced by the subsequent imbalance in the calcium metabolism. Alterations of skeletal properties occur as a result of disturbances in the normal mechanical loading environment of bone. These alterations or "adaptations" obey physical laws, but the precise mathematical relationships remain to be determined. Principles governing unloading and overloading of bone are gaining more attention as a consequence of the planning of manned space stations, moon and Mars bases and spaceflights of long duration. This paper reviews the subject of bone remodeling and presents a mathematical framework which allows for the prediction of skeletal adaptation on Earth and in non-Earth gravity environments by power law relationships.

Adaptation, Physiological↗

Automatic fixation facility for plant seedlings in the TEXUS Sounding Rocket Programme.

Automatic chemical fixation of plant seedlings within a 6 min period of reduced gravity (10(-4)g) was performed on three ballistic rocket flights provided by the German Sounding Rocket Programme TEXUS (Technologische Experimente unter Schwerelosigkeit = Technological Experiments in Microgravity). The described TEXUS experiment module consists of a standard experiment housing with batteries, cooling and heating systems, timer, and a data recording unit. Typically, 60 min before launch an experiment plug-in unit containing chambers with the plant material, the fixation system, and the temperature sensors is installed into the module which is already integrated in the payload section of the sounding rocket (late access). During the ballistic flight plant chambers are rapidly filled at pre-selected instants to preserve the cell structure of gravity sensing cells. After landing the plant material is processed for transmission electron microscopy. Up to now three experiments were successfully performed with cress roots (Lepidium sativum L.). Detailed improvements resulted in an automatic fixation facility which in principle can be used in unmanned missions.

Acceleration↗

[ECG changes during [correction of dring] high G centrifuge training].

ECG of 47 healthy subjects, during G-training on the human centrifuge were recorded and analyzed. Training consisted of 4 G profiles and carried out in 6 d. Subjects were divided into 3 groups according to different anti-G equipment used. The results showed that: high incidence of arrhythmia (87%) was observed, and it increased with the increase of G level; subjects with R/S < 1 under +Gz environment, or HR sharply dropped immediately after the +Gz load have lower G-tolerance; incidence of arrhythmia is lower when advanced anti-G equipment was used.

Acceleration↗

Non-invasive blood pressure measurement under G.

A new commercially available device (Finapres) which allows the continuous monitoring of blood pressure has been evaluated against measurements taken intra-arterially. It was found that Finapres tends to underestimate both systolic and diastolic blood pressure and differs significantly from, but correlates well, with intra-arterial line measurements. In general, the slope of the relationship between Finapres and intra-arterial values is close to unity. It is important that the arm is supported in a way that prevents any local tissue compression in the forearm. Finapres follows the changes in blood pressure induced by exposure to +Gz well, but may not give reliable absolute values. This device is able to give useful information on blood pressure in subjects exposed to +Gz on the man carrying centrifuge, with a minimum of discomfort.

Aerospace Medicine↗

Development of gravity-sensing organs in altered gravity.

Experiments are described in which the development of the gravity-sensing organs was studied in newt larvae reared in microgravity on the IML-2 mission and in Aplysia embryos and larvae reared on a centrifuge at 1 to 5 g. In Aplysia embryos, the statolith (single dense mass on which gravity and linear acceleration act) was reduced in size in a graded fashion at increasing g. In early post-metamorphic Aplysia or even in isolated statocysts from such animals, the number of statoconia produced is reduced at high g. Newt larvae launched before any of the otoconia were formed and reared for 15 days in microgravity had nearly adult labyrinths at the end of the IML-2 mission. The otoliths of the saccule and utricle were the same size in flight and ground-reared larvae. However, the system of aragonitic otoconia produced in the endolymphatic sac in amphibians was much larger and developed earlier in the flight-reared larvae. At later developmental stages, the aragonitic otoconia enter and fill the saccule. One flight-reared larva was maintained for nine months post-flight and the size of the saccular otolith, as well as the volume of otoconia within the endolymphatic sac, were considerably larger than in age-matched, ground-reared newts. This suggests that rearing in microgravity initiates a process that continues for several months after introduction to 1-g, which greatly increases the volume of otoconia. The flight-reared animal had abnormal posture, pointing its head upward, whereas normal ground-reared newts always keep their head horizontal. This suggests that rearing for even a short period in microgravity can have lasting functional consequences in an animal subsequently reared in 1-g conditions on Earth.

Animals↗