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PubMed2006

Effects of low-shear modeled microgravity on cell function, gene expression, and phenotype in Saccharomyces cerevisiae.

Only limited information is available concerning the effects of low-shear modeled microgravity (LSMMG) on cell function and morphology. We examined the behavior of Saccharomyces cerevisiae grown in a high-aspect-ratio vessel, which simulates the low-shear and microgravity conditions encountered in spaceflight. With the exception of a shortened lag phase (90 min less than controls; P < 0.05), yeast cells grown under LSMMG conditions did not differ in growth rate, size, shape, or viability from the controls but did differ in the establishment of polarity as exhibited by aberrant (random) budding compared to the usual bipolar pattern of controls. The aberrant budding was accompanied by an increased tendency of cells to clump, as indicated by aggregates containing five or more cells. We also found significant changes (greater than or equal to twofold) in the expression of genes associated with the establishment of polarity (BUD5), bipolar budding (RAX1, RAX2, and BUD25), and cell separation (DSE1, DSE2, and EGT2). Thus, low-shear environments may significantly alter yeast gene expression and phenotype as well as evolutionary conserved cellular functions such as polarization. The results provide a paradigm for understanding polarity-dependent cell responses to microgravity ranging from pathogenesis in fungi to the immune response in mammals.

Biomechanical Phenomena↗
PubMedDate not supplied

Effectiveness of centrifuge-induced artificial gravity with ergometric exercise as a countermeasure during simulated microgravity exposure in humans.

To test the effectiveness of centrifuge-induced artificial gravity with ergometric exercise, 12 healthy young men (20.7 +/- 1.9 yr) were exposed to simulated microgravity for 14 days of -6 degrees head-down bedrest. Half the subjects were randomly selected and loaded 1.2 G artificial gravity with 60 W (four out of six subjects) or 40 W (two out of six subjects) of ergometric workload on days 1, 2, 3, 5, 7, 9, 11, 12, 13, 14 (CM group). The rest of the subjects served as the control. Anti-G score, defined as the G-load x running time to the endpoint, was significantly elongated by the load of the centrifuge-ergometer. Plasma volume loss was suppressed (-5.0 +/- 2.4 vs. -16.4 +/- 1.9%), and fluid volume shift was prevented by the countermeasure load. Elevated heart rate and muscle sympathetic nerve activity after bedrest were counteracted, and exaggerated response to head-up tilt was also suppressed. Centrifuge-induced artificial gravity with exercise is effective in preventing cardiovascular deconditioning due to microgravity exposure, however, an effective and appropriate regimen (magnitude of G-load and exercise workload) should be determined in future studies.

Adult↗
PubMed2003

[Comparative evaluation of several methods preventing orthostatic disorders during simulation of the end-of-space-mission factors].

A new method for mitigation of the negative consequences of blood shift toward the cranial end as a result of simulation of the hemodynamic effects of microgravity (head-down tilt at -6 degrees) combines the lower body negative pressure (LBNP) and negative pressure respiration (NPR) to accelerate blood outflow from cerebral vessels and, additively, to restrain blood inflow from the lower body. The longitudinal hydrostatic pressure gradient is thus reproduced but main hemodynamic parameters assume values characteristic of the vertical posture. The objective was to compare the strength of LBNP and LBNP + NRP training in preventing orthostatic disorders following 7-d HDT. Subjects were 6 male volunteers aged 24 to 40 yrs. In the control experiment, every subject experienced episodes of orthostatic disorders. In the second experiment (LBNP), 4 subjects were afflicted; in the third experiment (LBNP + NPR), orthostatic disorders were minor, if at all. Absence of profound hemodynamic, autonomic or pre-syncope symptoms leads to the conclusion that combination LBNP + NPR is a more favorable preventive method than discrete LBNP.

Adult↗
PubMed2001

The neurogenic constrictor response of isolated rat saphenous artery is reduced after 2-week tail suspension.

Under real or simulated microgravity conditions the control of arterial vascular tone is greatly disturbed. The low arterial vessel reactivity to sympathetic influences may be the cause of an increase in flow in hind limb skeletal muscles in tail-suspended (TS) rats. Our previous experiments with constant pressure perfusion of rat hind limb demonstrated the reduced vasoconstrictor responses to sympathetic nerve stimulation in TS rats. Responses to exogenous noradrenaline depended on the perfusion conditions. It is known that the vessels of various branching orders noticeably differ in nerve density and in sensitivity to vasoconstrictor agonists. So under neurogenic or exogenous noradrenaline influences the vascular resistance may be increased at different levels of vascular bed, thus making the data analysis seriously complicated. This uncertainty may be overcome by investigation of a single vessel isolated from hind limb vascular bed. The saphenous artery, a resistance artery with dense innervation, is a very convenient object for this purpose. Thus, this study was aimed at comparing the effects of 2-week tail suspension upon the constrictor responses of isolated saphenous artery to neurogenic and exogenous noradrenaline stimuli in rats.

Animals↗
PubMed2004

[Male fertility in space].

The effects of simulated microgravity on mammalian reproduction were studied using tail-suspended mice, during parabolic flight in a jet plane and in a horizontal clinostat device. In the tail-suspended mouse, which is a model of the shift of body fluid in a microgravity environment, atrophy of the testis and decrease of serum testosterone level were observed. The sperm motility decreased in the muG condition produced by the parabolic flight. There were no statistically significant differences in the efficiency of achieving normal fertilization in vitro, but there was a statistically significant decrease in the number of embryos reaching the morula and blastocyst stages after 96 hours in culture under clinostat rotation. These results suggest that the process of fertilization in vitro is not sensitive to the gravitational vector. However, the possibility exists that microgravity increases the disturbance of spermatogenesis and sperm motility or the frequency of early embryonic lethality. On the possibility of reproduction in space, there are some problems such as the cosmic radiation besides gravity. It is necessary to carry out further experiments in outer space.

Animals↗
PubMed2006

Effects of acute simulated microgravity on nocturnal sleep, daytime vigilance, and psychomotor performance: comparison of horizontal and 6 degrees head-down bed rest.

This study examined the effect of acute simulated microgravity on nocturnal sleep, daytime vigilance, and psychomotor performance. Each of 7 volunteers were maintained for 3 days of head-down and horizontal bed rest in a counter-balanced design. Assessment measures were polysomnographic recordings on the first night and performance on psychophysiological tasks on the second day involving subjective and objective vigilance, P300, simple reaction time tasks, and dual performance tasks. No clear difference in sleep structure was observed between the head-down and horizontal conditions, except for a slight decrease in Stage 4 for head-down. Both subjective and objective daytime vigilance, P300, and the simple RT task showed no statistical difference, although tracking performance on the dual task showed deterioration at 10:00 for the head-down condition. These results suggest that nocturnal sleep, daytime vigilance, and psychophysiological functions were not disturbed in head-down sleep conditions, although there was a mild deterioration of higher attentional function in the morning.

Adult↗
PubMed1994

Aortic baroreflex control of heart rate after 15 days of simulated microgravity exposure.

To determine the effects of simulated microgravity on aortic baroreflex control of heart rate, we exposed seven male subjects (mean age 38 +/- 3 yr) to 15 days of bed rest in the 6 degrees head-down position. The sensitivity of the aortic-cardiac baroreflex was determined during a steady-state phenylephrine-induced increase in mean arterial pressure combined with lower body negative pressure to counteract central venous pressure increases and neck pressure to offset the increased carotid sinus transmural pressure. The aortic-cardiac baroreflex gain was assessed by determining the ratio of the change in heart rate to the change in mean arterial pressure between baseline conditions and aortic baroreceptor-isolated conditions (i.e., phenylephrine + lower body negative pressure + neck pressure stage). Fifteen days of head-down tilt increased the gain of the aortic-cardiac baroreflex (from 0.45 +/- 0.07 to 0.84 +/- 0.18 beats.min-1.mmHg-1; P = 0.03). Reductions in blood volume and/or maximal aerobic capacity may represent the underlying mechanism(s) responsible for increased aortic baroreflex responsiveness after exposure to a ground-based analogue of microgravity.

Adult↗
PubMed2002

MAP and src kinases control the induction of AP-1 members in response to changes in mechanical environment in osteoblastic cells.

The activating protein-1 (AP-1) complex plays a critical role in bone physiology, including its response to strain. We studied gene expression and nuclear translocation kinetics of the seven AP-1 members, after substrate deformation (Flexcell) or simulated microgravity (Clinostat), in osteoblastic ROS17/2.8 cells. Gene expression and nuclear translocation of all the AP-1 members were induced, under both conditions, with differences in their kinetics, except fosB mRNA in the Clinostat. Downregulation of protein kinase C (PKC) and COX1/2 or inhibition of ERK1/2, p38(MAPK) or src kinases had no major effect on AP-1 mRNA expression in the Flexcell. In contrast, ERK1/2, p38(MAPK) and src kinases treatment blocked nuclear translocation of almost all the AP-1 members in both models, except Fra-1, JunD after deformation and Fra-1, JunB after clinorotation. Thus, changes in the osteoblastic mechanical environment induced a dramatic induction of most of the AP-1 members with specific kinetics and involved MAPK and src kinase pathways, which differed whether the cells were stretched or clinorotated.

Active Transport, Cell Nucleus↗
PubMed2004

The microgravity environment for experiments on the International Space Station.

Experiments are sent to space laboratories in order to take advantage of the low-gravity environment. However, it is crucial to appreciate the distinction between the real microgravity environment and "weightlessness" or "simulated microgravity". The microgravity in space laboratories may be of much smaller magnitude than the gravitational acceleration on earth. However, it is not zero, nor even one microg (defined as 1e-6 earth gravity). Moreover, the orientation is not uniaxial, as on earth. The net acceleration that acts on a space experiment arises from, e.g., orbital mechanics, atmospheric drag, and thruster firings, and it can act on the experiments in gravity-like ways. In essence, a well-defined, stable 1 g acceleration on the earth's surface is substituted for a complex array of dynamically changing accelerations with ever-changing frequency content, magnitude and direction. This paper will show measured accelerations on the Shuttle from launch to orbit, as well as the latest measurements on the International Space Station (ISS). The ISS data presented here represent over 34,790 hours of data obtained from June 2002 to April 2003 during Increments 5 and 6 of the ISS construction cycle. The quasisteady acceleration level on the ISS has been measured to be on the order of a few microg during time allotted to microgravity mode. The vibratory acceleration environment spans a rich spectrum from 0.01-300 Hz.

Acceleration↗
PubMed2003

Chiral and non-chiral nutations in Arabidopsis roots grown on the random positioning machine.

Arabidopsis thaliana roots grown on a vertically set plate do not elongate straight down the gravitational vector, but by making waves and coils, and by conspicuously slanting towards the right-hand. This behaviour, in a previous paper, was ascribed to the simultaneous effect of three processes: circumnutation, positive gravitropism and negative thigmotropism. However, when the plants are grown on the Random Positioning Machine (RPM), in conditions that are believed to simulate space microgravitational conditions closely, the roots do not show the usual pattern. In the wild type, the roots make large loops to the right-hand side, whereas in the gravitropic and auxinic mutants aux1, eir1, rha1, they just move randomly around the initial direction. Therefore, if the movements made on the RPM are those produced by the exclusion of gravitropism and negative thigmotropism, as is apparent, the conclusion is that Arabidopsis roots are animated by a form of chiral circumnutation, that is lacking in the auxinic and gravitropic mutants aux1, eir1 and rha1. In addition, the 1 g condition appears to reduce the scatter among the circumnutating tracks produced by the roots of the wild types, but not among those of the mutants. Because there is a scarcity of literature regarding circumnutation in roots, it is not known how widely root chiral circumnutation is spread, but it is known that, in some previously studied species, just random nutations are observed. Two kinds of nutating movements seem to exist in plant roots and, whereas the random process does not seem to be connected with auxin physiology and transport, the chiral process appears to be connected in the same way as gravitropism is.

Arabidopsis↗
PubMed2000

Baroreflex control of muscle sympathetic nerve activity after 120 days of 6 degrees head-down bed rest.

To examine how long-lasting microgravity simulated by 6 degrees head-down bed rest (HDBR) induces changes in the baroreflex control of muscle sympathetic nerve activity (MSNA) at rest and changes in responses of MSNA to orthostasis, six healthy male volunteers (range 26-42 yr) participated in Valsalva maneuver and head-up tilt (HUT) tests before and after 120 days of HDBR. MSNA was measured directly using a microneurographic technique. After long-term HDBR, resting supine MSNA and heart rate were augmented. The baroreflex slopes for MSNA during Valsalva maneuver (in supine position) and during 60 degrees HUT test, determined by least-squares linear regression analysis, were significantly steeper after than before HDBR, whereas the baroreflex slopes for R-R interval were significantly flatter after HDBR. The increase in MSNA from supine to 60 degrees HUT was not different between before and after HDBR, but mean blood pressure decreased in 60 degrees HUT after HDBR. In conclusion, the baroreflex control of MSNA was augmented, whereas the same reflex control of R-R interval was attenuated after 120 days of HDBR.

Adult↗
PubMed2000

Exploring dynamic similarity in human running using simulated reduced gravity.

The Froude number (a ratio of inertial to gravitational forces) predicts the occurrence of dynamic similarity in legged animals over a wide range of sizes and velocities for both walking and running gaits at Earth gravity. This is puzzling because the Froude number ignores elastic forces that are crucial for understanding running gaits. We used simulated reduced gravity as a tool for exploring dynamic similarity in human running. We simulated reduced gravity by applying a nearly constant upward force to the torsos of our subjects while they ran on a treadmill. We found that at equal Froude numbers, achieved through different combinations of velocity and levels of gravity, our subjects did not run in a dynamically similar manner. Thus, the inertial and gravitational forces that comprise the Froude number were not sufficient to characterize running in reduced gravity. Further, two dimensionless numbers that incorporate elastic forces, the Groucho number and the vertical Strouhal number, also failed to predict dynamic similarity in reduced-gravity running. To better understand the separate effects of velocity and gravity, we also studied running mechanics at fixed absolute velocities under different levels of gravity. The effects of velocity and gravity on the requirements of dynamic similarity differed in both magnitude and direction, indicating that there are no two velocity and gravity combinations at which humans will prefer to run in a dynamically similar manner. A comparison of walking and running results demonstrated that reduced gravity had different effects on the mechanics of each gait. This suggests that a single unifying hypothesis for the effects of size, velocity and gravity on both walking and running gaits will not be successful.

Animals↗
PubMed2005

Blood volume measurement: The comparison of pulse dye densitometry and Dill and Costill's methods.

In many clinical situations, it is crucial to determine circulating blood volume (BV) easily and to repeat this measurement. The Dye DensitoGram Analyzer (DDG, Nihon Kohden Corp) measures semi-automatically BV, using an injection of IndoCyanine Green (ICG, 10 mg), and avoiding intermittent blood samples. The DDG was used during a 90-day microgravity simulation by Head-Down-Tilt bed rest (HDT) to measure BV and compared with the calculation of the plasma volume (PV) variations according to Dill and Costill's formula (DC). Seventeen healthy volunteers were included: 8 control subjects (Co) and 9 subjects submitted to a resistive exercise counter-measure (CM). Measurements were performed, one day before HDT, on days 3 and 90 of HDT and on day 9 after HDT. A double measurement of the BV was performed to assess the repeatability of this method. On the last day of HDT a significant decrease (p < 0.05) in the PV was noted with the DDG (Co: -12.3+/-5.7%, CM: -9.0+/-5.3%) and DC; (Co: -4.7+/-1.8%, CM: -6.8+/-2.5%). A good repeatability of the technique was shown with a low intrasubjects coefficient of variation (4.95+/-0.95%) and an acceptable intersubjects coefficient of variation (15.30+/-1.13%). No correlation was noted between DDG and DC (r2 = 0.27). The DDG gives a good repeatability, not affected by the microgravity exposure. Thanks to its capacity to measure accurately the BV within 7-10 min, this device presents major advantages for clinical use and research purpose.

Adult↗
PubMed1998

Formation and differentiation of three-dimensional rat marrow stromal cell culture on microcarriers in a rotating-wall vessel.

Using a high aspect ratio vessel (HARV), this study investigated the formation of 3-D rat marrow stromal cell culture on microcarriers and the expression of bone-related biochemical markers under conditions of simulated microgravity. In addition, it calculated the shear stresses imparted on the surface of microcarriers of different densities by the medium fluid in an HARV. Secondary rat marrow stromal cells were cultured on two types of microcarriers, Cytodex-3 beads and modified bioactive glass particles. Examination of cellular morphology by scanning electron microscopy revealed the presence of three-dimensional multicellular aggregates consisting of multiple cell-covered Cytodex-3 microcarriers bridged together. Mineralization was observed in the aggregates. Spherical cell-bead aggregates were observed in an HARV, while cell-bead assemblies were mostly loosely packed in a chain-like or branched structure in a cell bag. The expressions of alkaline phosphatase activity, collagen type I, and osteopontin were shown via the use of histochemical staining, immunolabeling, and confocal scanning electron microscopy. Using a numerical approach, it was found that at a given rotational speed and for a given culture medium, a larger density difference between the microcarrier and the culture medium (e.g., a modified bioactive glass particle) imparted a higher maximum shear stress on the microcarrier.

Alkaline Phosphatase↗
PubMed2004

Morphological changes in woody stem of Prunus jamasakura under simulated microgravity.

When the four-week-old woody stem of Prunus jamasakura was grown under simulated microgravity condition on a three-dimensional clinostat, it bent at growth, and width of its secondary xylem decreased due to the reduction of fiber cell numbers and a smaller microfibril angle in the secondary cell wall, as reported in our previous paper. Gravity induces the development of the secondary xylem that supports the stem upward against the action of gravity. In this study, morphological changes of the tissues and cells were microscopically observed. Disorder was found in the concentric structure of tissues that organize the stem. The radial arrangement of the cells was also disturbed in the secondary xylem, and in the secondary phloem secondary cell walls of the bast fiber cells were undeveloped. These findings suggest that differentiation and development of the secondary xylem and the bast fiber cells are strongly controlled by terrestrial gravity. These tissue and cells functions to support the stem under the action of gravity. Furthermore, clinorotation induced disorder in the straight joint of vessel elements and the lattice-like structure of radial parenchyma cells, which is responsible for water transportation and storage, respectively. Gravity is an essential factor for keeping the division and differentiation normal in woody stem.

Cell Wall↗
PubMed2003

Effects of strength training, using a gravity-independent exercise system, performed during 110 days of simulated space station confinement.

The efficacy of a resistance exercise paradigm, using a gravity-independent flywheel principle, was examined in four men subjected to 110 days of confinement (simulation of flight of international crew on space station; SFINCSS-99). Subjects performed six upper- and lower-body exercises (calf raise, squat, back extension, seated row, lateral shoulder raise, biceps curl) 2-3 times weekly during the confinement. The exercise regimen consisted of four sets of ten repetitions of each exercise at estimated 80-100% of maximal effort. Work was measured and recorded in each exercise session. Maximal voluntary isometric force in the calf press, squat and back extension, was assessed at three different joint angles before and after confinement. Overall, the training load (work) increased in all subjects (range 16-108%) over the course of the intervention. Maximal voluntary isometric force was unchanged following confinement. Although the perceived level of strain and comfort varied between exercises and among individuals, the results of the present study suggest this resistance exercise regimen is effective in maintaining or even increasing performance and maximal force output during long-term confinement. These findings should be considered in the design of resistance exercise hardware and prescriptions to be employed on the International Space Station.

Adaptation, Physiological↗
PubMed2006

Automating arm movement training following severe stroke: functional exercises with quantitative feedback in a gravity-reduced environment.

An important goal in rehabilitation engineering is to develop technology that allows individuals with severe motor impairment to practice arm movement without continuous supervision from a rehabilitation therapist. This paper describes the development of such a system, called Therapy WREX or ("T-WREX"). The system consists of an orthosis that assists in arm movement across a large workspace, a grip sensor that detects hand grip pressure, and software that simulates functional activities. The arm orthosis is an instrumented, adult-sized version of the Wilmington Robotic Exoskeleton (WREX), which is a five degrees-of-freedom mechanism that passively counterbalances the weight of the arm using elastic bands. After providing a detailed design description of T-WREX, this paper describes two pilot studies of the system's capabilities. The first study demonstrated that individuals with chronic stroke whose arm function is compromised in a normal gravity environment can perform reaching and drawing movements while using T-WREX. The second study demonstrated that exercising the affected arm of five people with chronic stroke with T-WREX over an eight week period improved unassisted movement ability (mean change in Fugl-Meyer score was 5 points +/- 2 SD; mean change in range of motion of reaching was 10%, p < 0.001). These results demonstrate the feasibility of automating upper-extremity rehabilitation therapy for people with severe stroke using passive gravity assistance, a grip sensor, and simple virtual reality software.

Adult↗
PubMed2001

Simulated microgravity, psychic stress, and immune cells in men: observations during 120-day 6 degrees HDT.

Because 6 degrees head-down tilt (HDT) is an established method to mimic low gravity on earth, the aim of the present study was to determine the effects of 120-day HDT on psychic stress and peripheral blood immune cells in six healthy male volunteers. Psychological state was assessed by a current stress test, and cortisol was measured in saliva. During HDT, all volunteers developed psychic stress, and the diurnal rhythm of cortisol secretion was significantly altered. In addition, urine excretion of dopamine and norepinephrine increased. The innate part of the immune response was activated, as evidenced by the increase in the expression of beta(2)-integrins on polymorphonuclear leukocytes and a rise in the number of circulating natural killer (NK) cell lymphocytes. The ratio of T-helper to T-cytotoxic and T-suppressor cells decreased, whereas no changes in T and B lymphocytes were observed. Plasma levels of interleukin-6 increased significantly and returned to basal levels after the end of the HDT period. Thus 6 degrees HDT appears to be a valid model to induce psychic stress and neuroendocrine-related changes in the immune system, changes that might also be encountered by astronauts and cosmonauts during long-duration spaceflights.

Adult↗