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PubMed1991

Effects of suspension-induced osteopenia on the mechanical behaviour of mouse long bones.

Whereas most studies of tail-suspension induced osteopenia have utilized rat femora, the present study investigated the effects of a 14 day tail-suspension on the mechanical behaviour of mice femora, tibiae and humeri. Force-deflection properties were obtained via three-point bending for long bones from suspended and control mice. Whole bone behaviour was characterized by converting the force-deflection values to stiffness, strength, ductility and energy parameters which were not normalized for specimen geometry. The effects of a systematic variation in the deflection rate over the range 0.1-10 mm min-1 were also evaluated. Statistical analysis indicated that the primary effect of the tail-suspension period was lowered bone mass which was manifested mechanically through lower values of the bone strength parameters. These effects were similar in the bones of both the fore and hind limbs. The results also demonstrated that the stiffness, ductility and energy characteristics were much less influenced by the tail-suspension. Whereas a significant dependence of the bone strength values upon deflection rate was observed for the femora and humeri, the other mechanical parameters were less sensitive. Based upon the nature of the physical and mechanical changes observed in the long bones following tail-suspension, the mouse appears to be a suitable animal model for the study of osteopenia.

Animals↗
PubMed1996

Induction of hydrotropism in clinorotated seedling roots of Alaska pea, Pisum sativum L.

Roots of the agravitropic pea (Pisum sativum L.) mutant ageotropum show positive hydrotopism, whereas roots of Alaska peas are hydrotropically almost non-responsive. When the gravitropic response was nullified by rotation on clinostats, however, roots of Alaska peas showed unequivocal positive hydrotropism in response to a water potential gradient. These results suggest that roots of Alaska peas possess normal ability to respond hydrotropicallly and their weak hydrotropic response results from a counteracting effect of gravitropism.

Gravitation↗
PubMed1996

Calbindins decreased after space flight.

Exposure of the body to microgravity during space flight causes a series of well-documented changes in Ca2+ metabolism, yet the cellular and molecular mechanisms leading to these changes are poorly understood. Calbindins, vitamin D-dependent Ca2+ binding proteins, are believed to have a significant role in maintaining cellular Ca2+ homeostasis. In this study, we used biochemical and immunocytochemical approaches to analyze the expression of calbindin-D28k and calbindin-D9k in kidneys, small intestine, and pancreas of rats flown for 9 d aboard the space shuttle. The effects of microgravity on calbindins in rats from space were compared with synchronous Animal Enclosure Module controls, modeled weightlessness animals (tail suspension), and their controls. Exposure to microgravity resulted in a significant and sustained decrease in calbindin-D28k content in the kidney and calbindin-D9k in the small intestine of flight animals, as measured by enzyme-linked immunosorbent assay (ELISA). Modeled weightlessness animals exhibited a similar decrease in calbindins by ELISA. Immunocytochemistry (ICC) in combination with quantitative computer image analysis was used to measure in situ the expression of calbindins in the kidney and the small intestine, and the expression of insulin in pancreas. There was a large decrease of immunoreactivity in renal distal tubular cell-associated calbindin-D28k and in intestinal absorptive cell-associated calbindin-D9k of space flight and modeled weightlessness animals compared with matched controls. No consistent difference in pancreatic insulin immunoreactivity between space flight, modeled weightlessness, and controls was observed. Regression analysis of results obtained by quantitative ICC and ELISA for space flight, modeled weightlessness animals, and their controls demonstrated a significant correlation. These findings after a short-term exposure to microgravity or modeled weightlessness suggest that a decreased expression of calbindins may contribute to the disorders of Ca2+ metabolism induced by space flight.

Animals↗
PubMed1999

A spaceflight experiment for the study of gravimorphogenesis and hydrotropism in cucumber seedlings.

Seedlings of Cucurbitaceae plants form a protuberance, termed peg, on the transition zone between hypocotyl and root. Our spaceflight experiment verified that the lateral positioning of a peg in cucumber seedlings is modified by gravity. It has been suggested that auxin plays an important role in the gravity controlled positioning of a peg on the ground. Furthermore, cucumber seedlings grown in microgravity developed a number of the lateral roots that grew towards the water containing substrate in the culture vessel, whereas on the ground they oriented perpendicular to the primary root growing down. The response of the lateral roots in microgravity was successfully mimicked by clinorotation of cucumber seedlings on the three dimensional clinostat. However, this bending response of the lateral roots was observed only in an aeroponic culture of the seedlings but not in solid medium. We considered the response of the lateral roots in microgravity and on clinostat as positive hydrotropism that could easily be interfered by gravitropism on the ground. This system with cucumber seedlings is thus a useful model of spaceflight experiment for the study of the gravimorphogenesis, root hydrotropism and their interaction.

Cucumis sativus↗
PubMed2004

Is perception of upper body orientation based on the inertia tensor? Normogravity versus microgravity conditions.

During lateral leg raising, a synergistic inclination of the supporting leg and trunk in the opposite direction to the leg movement is performed in order to preserve equilibrium. As first hypothesized by Pagano and Turvey (J Exp Psychol Hum Percept Perform, 1995, 21:1070-1087), the perception of limb orientation could be based on the orientation of the limb's inertia tensor. The purpose of this study was thus to explore whether the final upper body orientation (trunk inclination relative to vertical) depends on changes in the trunk inertia tensor. We imposed a loading condition, with total mass of 4 kg added to the subject's trunk in either a symmetrical or asymmetrical configuration. This changed the orientation of the trunk inertia tensor while keeping the total trunk mass constant. In order to separate any effects of the inertia tensor from the effects of gravitational torque, the experiment was carried out in normo- and microgravity. The results indicated that in normogravity the same final upper body orientation was maintained irrespective of the loading condition. In microgravity, regardless of loading conditions the same (but different from the normogravity) orientation of the upper body was achieved through different joint organizations: two joints (the hip and ankle joints of the supporting leg) in the asymmetrical loading condition, and one (hip) in the symmetrical loading condition. In order to determine whether the different orientations of the inertia tensor were perceived during the movement, the interjoint coordination was quantified by performing a principal components analysis (PCA) on the supporting and moving hips and on the supporting ankle joints. It was expected that different loading conditions would modify the principal component of the PCA. In normogravity, asymmetrical loading decreased the coupling between joints, while in microgravity a strong coupling was preserved whatever the loading condition. It was concluded that the trunk inertia tensor did not play a role during the lateral leg raising task because in spite of the absence of gravitational torque the final upper body orientation and the interjoint coupling were not influenced.

Adult↗
PubMed2000

Human orientation and movement control in weightless and artificial gravity environments.

Our goal is to summarize what has been learned from studies of human movement and orientation control in weightless conditions. An understanding of the physics of weightlessness is essential to appreciate the dramatic consequences of the absence of continuous contact forces on orientation and posture. Eye, head, arm, leg, and whole body movements are discussed, but only experiments whose results seem relatively incontrovertible are included. Emphasis is placed on distinguishing between virtually immediate adaptive compensations to weightlessness and those with longer time courses. The limitations and difficulties of performing experiments in weightless conditions are highlighted. We stress that when astronauts and cosmonauts return from extended space flight they do so with both physical "plant" and neural "controller" structurally and functionally altered. Recent developments in adapting humans to artificial gravity conditions are discussed as a way of maintaining sensory-motor and structural integrity in extended missions involving transitions between different force environments.

Humans↗
PubMed2002

Effect of human head flexion on the control of peripheral blood flow in microgravity and in 1 g.

This study evaluated, in six healthy subjects, whether head flexion, which stimulates the vestibular system and the tonic neck receptors, interferes with cardiovascular regulation. Arterial parameters were measured continuously using a pulsed Doppler ultrasound probe during parabolic flights with subjects either in the supine craned-head position (control) or in the supine anterior neck flexion bent-neck position. Exposure to 0 g induced a fluid shift towards the head (stroke volume +8%, P<0.05). Compared to the control situation the mean (SD) blood flow in the femoral artery decreased [ -10 (9)% vs +1 (10)%; P<0.05], and the ratio cerebral artery:femoral artery blood flow ( : ) increased [+8 (14)% vs -4 (7)%; P<0.05], in the bent-neck position. Thus, neck flexion without otolith loading (subject in 0 g) favoured cerebral perfusion during the exposure to 0 g. The return to 1 g, even in the supine position, induced a fluid shift towards the lower limbs. From 0 to 1 g, reduced less [ +6 (8)% vs -1 (8)%; P<0.05], and the : decreased more [-11 (9)% vs 0 (10)%; P<0.05], in the bent-neck position than in the control position. Thus the redistribution of peripheral blood flow in response to the fluid shift towards the legs was less efficient in the bent-neck position. In 0 g environment the passive flexion of the neck (neck receptor stimulation only) increased resistance in the femoral artery [ R(fa) +20 (21)%; P<0.05] and reduced the [-15(10)%; P<0.07] which increased the redistribution of flow towards the brain [; +12 (7)%; P<0.07]. This response was of lower amplitude when both otoliths and neck muscle were stimulated (neck flexion in 1 g) [ R(fa)+9 (7)%, P<0.05; -9 (12), NS; : 0 (12), NS]. We suggest that otolith and neck muscle stimulation (by neck flexion) trigger opposite vascular effects in response to a fluid shift towards the legs.

Adaptation, Physiological↗
PubMed2003

Parasympathetic heart rate modulation during parabolic flights.

During parabolic flight short periods of microgravity and hypergravity are created. These changes influence cardiovascular function differently according to posture. During the 29th parabolic flight campaign of the European Space Agency (ESA), the electrocardiogram (ECG) was recorded continuously in seven healthy volunteers in two positions (standing and supine). Five different phases were differentiated: 1 g (1 g=9.81 m/s(2)) before and after each parabola, 1.8 g at the ascending leg of the parabola (hypergravity), 0 g at the apex, 1.6 g at the descending leg (hypergravity). We assessed heart rate variability (HRV) by indices of temporal analysis [mean RR interval (meanRR), the standard deviation of the intervals (SDRR), and the square root of the mean squared differences of successive intervals (rMSSD) and coefficient of variation (CV)]. In the supine position no significant differences were shown between different gravity phases for all HRV indices. In the standing position the 0 g phase showed a tendency towards higher values of meanRR compared to the control and to the other phases ( p=NS). SDRR, rMSSD and CV were significantly higher compared to control ( p<0.05). Significantly higher values for meanRR in the supine position at 1 g and hypergravity ( p<0.05) were found when compared to standing. SDRR was significantly higher at 0 g in the standing position compared to supine [95 (44) ms vs. 50 (15) ms; p<0.05] and lower in other phases. rMSSD and CV showed the same trend ( p=NS). We confirm that, during parabolic flights, position matters for cardiovascular measurements. Time domain indices of HRV during different gravity phases showed: (1) higher vagal modulation of the autonomic nervous system in microgravity, when compared with normo- or hypergravity in standing subjects; and (2) no differences in supine subjects between different g phases.

Adaptation, Physiological↗
PubMed2004

Human adductor muscles atrophy after short duration of unweighting.

The purpose of this study was to investigate the effect of 20 days of bed rest (BR) on the physiological cross-sectional area (PCSA) of the adductor (AD) muscle group. Five healthy men and five healthy women were kept on a horizontal bed for 20 days. To calculate the PCSA of the AD muscle group, transaxial magnetic resonance imaging of the right thigh was acquired four times for each of the subjects: (1) pre BR, (2) on the tenth day of BR, (3) post BR, and (4) 1 month after BR. Overall, the PCSA measurements of the AD muscle group and its individual muscles were significantly decreased on the tenth day of BR and at post BR; however, they had recovered to the baseline within 1 month of the reambulation period. The amount and pattern of relative change in the PCSA measurements of the AD muscle group were similar to those of the knee extensor (KE) and knee flexor (KF) muscle groups studied previously. These results suggested that the AD muscle group showed an atrophic response that was similar to the KE and KF muscle groups after unweighting; therefore, we should pay much more attention to the AD muscle group when considering countermeasures for future human spaceflight projects.

Adult↗
PubMed1998

Effects of chronic low frequency stimulation on contractile and elastic properties of hindlimb suspended rat soleus muscle.

Chronic low frequency stimulation (10 Hz, 8 h x day[-1]) was used in this study to prevent the changes in the contractile and elastic properties of rat soleus muscles induced by 3 weeks of hindlimb suspension (HS). Results showed that electrostimulation was able to counteract in part the decrease in soleus muscle mass and tension output induced by unweighting. On the other hand, the increases in maximal shortening velocity and twitch speed following HS were not prevented by stimulation. Unweighting was responsible for an increase in series elastic compliance of soleus muscle. Chronic stimulation successfully counteracted this increase in series compliance probably by changing the properties of the tendon. The partial recovery of muscle mass and tension output as a result of stimulation enhanced the role of contractile activity in preventing muscle atrophy. Moreover, the inefficiency of the tonic activity imposed by stimulation in preventing the increase in twitch speed of soleus muscle during HS demonstrated the primacy of neuronal activity. Discrepant results concerning changes in contraction kinetics deduced from the twitch could have been due to the fact that such myograms also depend on the series compliance.

Animals↗
PubMed1998

Energy and substrate metabolism during a 42-day bed-rest in a head-down tilt position in humans.

Microgravity-induced changes in body composition (decrease in muscle mass and increase in fat mass) and energy metabolism were studied in seven healthy male subjects during a 42-day bed-rest in a head-down tilt (HDT) position. Resting energy expenditure (REE), fat and glucose oxidation were estimated by indirect calorimetry on days 0, +8 and +40 of the HDT period. Assessments were performed both in post-absorptive conditions and following two identical test meals given at 3-h intervals. Body composition (dual x-ray absorptiometry) was measured on days 0, +27, +42. Mean post-absorptive lipid oxidation decreased from 53 (SEM 8) mg x min(-1) (day 0) to 32 (SEM 10) mg x min(-1) (day 8, P = 0.04) and 36 (SEM 8) mg x min(-1) (day 40, P = 0.06). Mean post-absorptive glucose oxidation rose from 126 (SEM 15) mg x min(-1) (day 0) to 164 (SEM 14) mg x min(-1) (day 8, P = 0.04) and 160 (SEM 20) mg x min(-1) (day 40, P = 0.07). Mean fat-free mass (FFM) decreased between days 0 and 42 [58.0 (SEM 1.8) kg and 55.3 (SEM 1.7) kg, P < 0.01] while fat mass increased without reaching statistical significance. The mean REE decreased from 1688 (SEM 50) kcal x day(-1) to 1589 (SEM 42) kcal x day(-1) (P = 0.056). Changes in REE were accounted for by changes in FFM. Mean energy intake decreased from 2532 (SEM 43) kcal x day(-1) to 2237 (SEM 50) kcal x day(-1) (day 40, P < 0.01) with only a minor decrease in the proportion of fat. We concluded that changes in fat oxidation at the whole body level can be found during HDT experiments. These changes were related to the decrease in FFM and could have promoted positive fat balance hence an increase in fat mass.

Adult↗
PubMed1999

The rat suspension model is also a good tool for inducing muscle hyperactivity.

The aim of the present study was to investigate changes in the mechanical characteristics of the rat epitrochlearis muscle as a result of a period of hyperactivity. A tail suspension model was used to impose postural activity in the forelimb musculature with the intention of inducing a relative slowness in the fast epitrochlearis muscle. A method of dual-controlled releases was used to obtain force-velocity and tension-extension relationships characterizing muscle mechanics. Results showed that: (1) mechanical characteristics are that of a fast-twitch muscle and (2) suspension induces a decrease in the maximal shortening velocity and a decrease in the compliance of series elastic elements. Changes in fibre type proportions are consistent with these mechanical adaptations. This demonstrates that an usual model of muscle hypoactivity can also be used for imposing hyperactivity of a postural nature, inducing muscle transformations towards a slower twitch muscle.

Animals↗
PubMed2006

Microgravity-related changes in gene expression after short-term exposure of Arabidopsis thaliana cell cultures.

Cell cultures of Arabidopsis thaliana (cv. Columbia) were used to screen for early alterations in gene expression as a response to altered gravitational fields. Genes of interest were selected from a larger group whose expression was altered under hypergravity (microarray study; M. Martzivanou and R. Hampp, Physiol. Plant. 118: 221-231, 2003). Transcriptional changes of these genes were studied within a time frame of up to 10 min of exposure to microgravity in a sounding-rocket experiment, to clinorotation (random positioning machine), and to hypergravity (8 g). We could identify a set of nine genes (mainly components of signaling chains) with increased transcript levels after about 6 min of exposure to microgravity. As clinorotation and hypergravity treatment did not alter the respective transcript amounts, we assume that the identified genes could be involved in a microgravity-related response.

Arabidopsis↗
PubMed2006

Cytoskeleton changes and impaired motility of monocytes at modelled low gravity.

Investigations performed in space have shown that gravity changes affect important cellular mechanisms like proliferation, differentiation, genetic expression, cytoskeletal architecture, and motility in lymphocytes, monocytes, and other mammalian cells. In particular, a dramatic depression of the mitogenic in vitro activation of human peripheral blood lymphocytes was observed at low gravity. The hypothesis of the present work is that a reduced interaction between T lymphocytes and monocytes, essential for the second signalling pathway, might be one of the reasons for the observed depression of the in vitro activation of human lymphocytes. Cell motility and with it a continuous rearrangement of the cytoskeletal network within the cell is essential for cell-to-cell contacts. Whereas nonactivated lymphocytes in suspension are highly motile at low gravity, no data are available so far on the motility of adherent monocytes. It thus can be argued that impaired monocyte locomotion and cytoskeletal changes could be responsible for a reduced interaction of monocytes with T lymphocytes. In this study, the locomotion ability of J-111 cells, an adherent monocyte cell line, attached to colloidal gold particles on coverslips and exposed to modelled low gravity in the random positioning machine was found to be severely reduced compared with that of controls and the structures of actin, tubulin, and vinculin were affected.

Actin Cytoskeleton↗
PubMed1997

Neonatal rat heart cells cultured in simulated microgravity.

In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two-dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by nonmyocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA-designed High-Aspect Ratio Vessel (HARV) bioreactors provide a low shear environment that allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells cultured in HARVs adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARVs using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar; however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissuelike organization of cardiac cells in vitro.

Animals↗
PubMed1997

Microgravity tissue engineering.

Tissue engineering studies were done using isolated cells, three-dimensional polymer scaffolds, and rotating bioreactors operated under conditions of simulated microgravity. In particular, vessel rotation speed was adjusted such that 10 mm diameter x 2 mm thick cell-polymer constructs were cultivated in a state of continuous free-fall. Feasibility was demonstrated for two different cell types: cartilage and heart. Conditions of simulated microgravity promoted the formation of cartilaginous constructs consisting of round cells, collagen and glycosaminoglycan (GAG), and cardiac tissue constructs consisting of elongated cells that contracted spontaneously and synchronously. Potential advantages of using a simulated microgravity environment for tissue engineering were demonstrated by comparing the compositions of cartilaginous constructs grown under four different in vitro culture conditions: simulated microgravity in rotating bioreactors, solid body rotation in rotating bioreactors, turbulent mixing in spinner flasks, and orbital mixing in petri dishes. Constructs grown in simulated microgravity contained the highest fractions of total regenerated tissue (as a percent of construct dry weight) and of GAG, the component required for cartilage to withstand compressive force.

Animals↗