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Effects of 5 weeks of lower limb suspension on muscle size and strength.

Lack of weight-bearing, as occurs in space, appears to be associated with reductions in strength and mass of skeletal muscle. Very limited data, however, is at hand describing changes in skeletal muscle size and function following manned space missions. Our current knowledge therefore is mainly based on studies of space flown rats. It is obvious though that this information, only in part can be extrapolated to humans. A few bed rest studies have demonstrated that decreases in strength and muscle size are substantial. At this time, however, the magnitude or time course of such changes either in response to space flight or simulations of microgravity have not been defined. In the last few years we have employed a human model to simulate unloading of lower limb skeletal muscles that occurs in microgravity. This model was essentially adopted from the rat hindlimb suspension technique. The purpose of this study was to assess the magnitude of decreases in muscle strength and size as a result of five weeks of unilateral lower limb suspension.

Adaptation, Physiological↗

Effects of fast clinostat treatment and microgravity on Vicia faba L. mesophyll cell protoplast ubiquitin pools and actin isoforms.

With parabolic rocket flights and fast clinostat treatments, the effect of microgravity on ubiquitin, ubiquitin-protein conjugates, and actin isoforms of Vicia faba mesophyll protoplasts was studied. Western immunoblotting with ubiquitin antibodies revealed that simulated and particularly, real microgravity influenced the amount of free ubiquitin and of 18, 19, and 40 kD ubiquitin conjugates by inducing strong oscillations in the proteins concentrations over time. Simulated microgravity and microgravity-phase during parabolic rocket flights resulted in a decrease of actin isoforms. Results obtained support the assumption, that microgravity and fast clinostat treatment have a direct effect on Vicia faba mesophyll protoplast metabolic activities.

Actins↗

[Impact of simulated microgravity on the expression and distribution of cardiac gap junction protein CX43].

OBJECTIVE: To observe the expression and distribution changes of connexin 43 (CX) in rats' myocardium after simulated microgravity and explore the partial mechanism of arrhythmia. METHOD: Male Wistar rats were randomly assigned to either tail-suspension group (SUS) or control group (CON). Immunohistochemistry and Western Blot were used to detect the expression and the distribution of CX43. Electromicroscope was used to observe the ultrastructural changes. RESULT: In SUS group, the decrease of CX43 and the distribution disturbance were obvious (P<0.05), the proportion of the side-to-side gap junction increased (P<0.05), and space between some gap junctions disappeared. CONCLUSION: The results show that CX43 decreased significantly and distributed irregularly after simulated microgravity. These can cause the changes of cardiac electric conduction velocity and direction. As a result, conduction block and reentry may occur. And these might be the partial mechanism of cardiac arrhythmia.

Animals↗

Renal osmoregulatory function during simulated space flight.

Experiments are described on the dynamics of the renal excretion of water and electrolytes, osmotically active substances and osmotically free water during bed rest. In short-term experiments (up to 10-14 days) renal changes are mainly associated with haemodynamic disturbances and fluid redistribution, whereas in longer duration experiments (up to 50 days) they are brought about by metabolic changes.

Adaptation, Physiological↗

Early renal changes in 45 degrees HDT rats.

BACKGROUND: Both microgravity and simulated microgravity models, such as the 45HDT (45 degrees head-down tilt), cause a redistribution of body fluids indicating a possible adaptive process to the microgravity stressor. Understanding the physiological processes that occur in microgravity is a first step to developing countermeasures to stop its harmful effects, i.e., (edema, motion sickness) during long-term space flights. HYPOTHESIS: Because of the kidneys' functional role in the regulation of fluid volume in the body, it plays a key role in the body's adaptation to microgravity. METHODS: Rats were injected intramuscularly with a radioactive tracer and then lightly anesthetized in order to facilitate their placement in the 45HDT position. They were then placed in the 45HDT position using a specially designed ramp (45HDT group) or prone position (control group) for an experimental time period of 1 h. During this period, the 99mTc-DTPA (technetium-labeled diethylenepentaacetate, MW=492 amu, physical half-life of 6.02 h) radioactive tracer clearance rate was determined by measuring gamma counts per minute. The kidneys were then fixed and sectioned for electron microscopy. A point counting method was used to quantitate intracellular spaces of the kidney proximal tubules. RESULTS: 45HDT animals show a significantly (p=0.0001) increased area in the interstitial space of the proximal tubules. CONCLUSIONS: There are significant changes in the kidneys during a 1 h exposure to a simulated microgravity environment that consist primarily of anatomical alterations in the kidney proximal tubules. The kidneys also appear to respond differently to the initial periods of head-down tilt.

Adaptation, Physiological↗

The Spacelab 3 simulation: basis for a model of growth plate response in microgravity in the rat.

Data from Spacelab 3 (SL3) suggested that spaceflight significantly reduces the activity of the rat tibial growth plate. Animal processing after SL3 began twelve hours post-landing, so data reflect post-flight re-adaptation in addition to spaceflight effects. To determine if a twelve-hour period of weight bearing after seven days of unloading could affect the physes of spaceflown rats, the present study assessed the growth plate response to unloading with or without a reloading period. Rats were subjected to hind-limb suspension for seven days and then euthanized, with or without twelve hours of reloading. Activity of the growth plate was assessed by morphometric analysis. Rats suspended without reloading had reserve zone (RZ) height greater than controls, and shorter hypertrophy/calcification zone (HCZ) with fewer cells. The greater RZ was associated with a larger cell area, indicating a possible mitotic delay or secretion defect. Twelve hours of reloading decreased RZ height and cell number, and restored the number of cells in HCZ to control values, but the number of cells in the proliferative zone and height in HCZ were reduced. These results suggest the rebound response to preserve/restore skeletal function after a period of unloading involves an acceleration of growth associated with a decreased cell cycle time in PZ. Changes during the reloading period in this simulation support our hypothesis that the effects of spaceflight on SL3 growth plates were altered by changes that occurred post-landing. The similarities in response to unloading by suspension or during spaceflight are used to propose a model of growth plate response during spaceflight.

Animals↗

[Effect of weightlessness on the functions of perception and the reproduction of gravitational vertical in response to optokinetic stimulation].

In weightlessness spatial orientation largely depends on coordinated interaction of sensory systems involved in static orientation. The purpose of the present study was to quantify perception of spatial coordinates in response to optokinetic stimulation. Effects of weightlessness were simulated by dry water immersion hypokinesia. A custom-made optokinetic stimulator equipped with a device to measure perception of the gravitational vertical in response to optokinetic stimulation was used. A special platform with rubber bungee cords was employed to determine the role of the proprioceptive afferent input. It was demonstrated that during immersion day 1 the angle of erroneous perception of the vertical line consistently increased. By immersion day 3 the angle diminished suggesting adaptive developments. Additional proprioceptive inflow during immersion reduced significantly the error of perception. These findings indicate that the proprioceptive afferent input plays an important role in the mechanisms of spatial orientation at an early stage of exposure to weightlessness.

Adult↗

Venous distensibility and venous emptying in the legs during a 42-day -6 degrees head-down bedrest.

Exposure to actual or simulated microgravity is known to result in changes in lower limb venous compliance or distensibility which may play a role in post-bedrest or postflight orthostatic intolerance. Venous deconditioning has only been described in terms of changes in vascular compliance or distensibility. But a complete understanding of changes in venous hemodynamics and cardiovascular regulation occurring under these conditions has to take into account changes in emptying capacities of the veins which influence venous return, cardiac filling, and cardiac output regulation. Moreover, few data are available about the course of changes in venous hemodynamics for periods of simulated microgravity longer than 4 weeks. The purpose of this investigation was to measure parameters of venous compliance and venous emptying before, during, and after a 42-day period of bedrest at -6 degrees head-down tilt for a better understanding of long term venous physiological adaptation to microgravity.

Adult↗

Bed rest immobilization with various oral sodium supply: plasma hormones and body fluids.

OBJECTIVE: To answer the question if plasma hormone concentrations (plasma renin activity--PRA, vasopressin--pAVP, and aldosterone concentration) due to antiorthostatic immobilization (8 days -6 degree head-down tilt bed rest--HDBR) are altered by oral salt load, we provided constant sodium supply during 4 days ambulatory conditions followed by 8 days HDBR in 10 normotensive men. METHODS: A low' (LS: 143 10 mM) and high' (HS: 434+/-17 mM Na+/d excreted) sodium treatment were provided in randomized order, separated > or = 1 mo. Before and at the end of HDBR, hemodynamic variables and thoracic impedance were determined, and blood was taken for aldosterone and PRA, venous hematocrit, and plasma mass density. Extracellular fluid volume and pAVP were determined every second day. Whole body electrical impedance spectroscopy was employed to assess changes in extracellular volume, hormone determinations were done with radioimmunoassay, mass density measurements with the mechanical oscillator technique. RESULTS: Extracellular volume decreased with HDBR (LS: -4.0%, p=0.002; HS: -5.8%, p=0.018) without significant difference between salt treatments. Resting hormone levels were not altered by HDBR, but pAVP was lower (5.5+/-0.1 pg/ml) in HS than in LS (7.2+/-0.3 pg/ml) as was plasma aldosterone (HS: 69+/-7 pg/ml, LS: 180+/-24 pg/ml). On the other hand, HDBR reduced extracellular volume by ?5% irrespective of dietary sodium supply. CONCLUSIONS: Our data support the hypothesis that hormonal activities are more affected by oral salt load than by simulated short-term space flight, and suggest that the reduction of extracellular fluid volume due to head down bed rest is not influenced by moderate changes of dietary sodium supply.

Administration, Oral↗

A proteolytic NH2-terminal truncation of cardiac troponin I that is up-regulated in simulated microgravity.

In a tail suspension rat model, we investigated changes in myofilament protein during cardiac adaptation in simulated microgravity. Contractile force and velocity of cardiac muscle were decreased in the tail suspension rats as compared with the control. Ca(2+)-dependent actomyosin ATPase activity was also decreased; however, sensitivity of cardiac muscle to Ca(2+) activation was unchanged. There was no change in expression of myosin heavy chain, tropomyosin, troponin T, or troponin I isoforms in hearts of tail suspension rats. A novel finding is a fragment of cardiac troponin I (cTnI) that had increased amounts in the heart of tail suspension rats. Binding of this cTnI fragment by a monoclonal antibody that specifically recognizes the COOH terminus indicates an intact COOH terminus. NH(2)-terminal sequence analysis of the cTnI fragment revealed truncations primarily of amino acids 1-26 and 1-27 and smaller amounts of 1-30, including Ser(23) and Ser(24), which are substrates of protein kinase A phosphorylation. This cTnI fragment is present in normal cardiac muscle and incorporated into myofibrils, indicating a role in regulating contractility. This proteolytic modification of cTnI up-regulated during simulated microgravity suggests a potential role of the NH(2)-terminal segment of cTnI in functional adaptations of cardiac muscle.

Actins↗

A NMR-compatible and reduced gravity simulation based (NRG) bioreactor for on-line monitoring cell culture metabolism.

We developed a NMR-compatible microgravity-based bioreactor (NRG[R]) that offers the advantage of an analytical non-invasive approach associated to the effects of an optimized suspension culture. The simulated microgravity conditions reached in the bioreactor are analogous to those of commercial apparatus like the Rotating Wall Vessel (RWV) system. The faster proliferation of endothelial cells cultured in the NRG bioreactor (doubling time : 28 +/- 1.7 vs. 43 +/- 5.6 h of the control grown in RWV) are attributed to different oxygenation conditions and medium wash out.

Bioreactors↗

Influence of simulated microgravity on the exercise performance of Fischer 344 rats.

Measurements from mission specialists after space flights or from subjects subjected to head down tilt experiments have demonstrated a decrease in exercise performance. Similar decreases have been reported for rats that have participated in simulated microgravity studies using the head down-tail suspended method of Morey-Holton (HDS). Because it is unclear whether older animal populations would exhibit similar responses, we undertook a HDS study with Fischer 344 male rats.

Aging↗

Bone demineralization mechanisms at level of free radicals and nanoscale subsystems of bone tissue.

Influence of simulated microgravity on characteristics of rat bones was investigated by electron paramagnetic resonance (EPR). For simulation of microgravity condition the hanging of animal by tail was used. The main measurements were performed for diaphysis of femoral bones. The quantity of native radicals Rn, localized in an organic matrix, and carbonate radicals CO2-, localized on a surface of bioapatite nanocrystals, were determined. The decrease of CO2- radical quantity in bones of experimental animals have shown, that due to simulation of microgravity the decrease of "collagen-nanocrystals" interaction takes place. It is shown that the EPR method open possibilities to receive the unique information about bone demineralization processes at level of free radicals and nanoscale subsystems of bone tissue.

Animals↗

A study of orientation in a zero gravity environment by means of virtual reality simulation.

When the International Space Station (ISS) is completed and starts its operation, crew members will be stationed for three months or more in orbit aboard the ISS. As they stay longer in the space environment, "habitability" for them will become most important in the design of the interior space. One of the problems about habitability in a zero gravity (0 G) environment is disorientation. Crew members have difficulty in discriminating between "up" and "down" and more serious disorientations may cause space motion sickness. Crew members rely on visual perception to orient themselves because they can't use their sense of equilibrium in a 0 G environment. Although color and the direction of equipment of Space Shuttles or modules has been considered, no systematic study has been conducted on interior space. This study intended to clarify how people acquire visual information and recognize their orientation in a 0 G environment by an experiment in which a subject wears a head-mounted display (HMD) and enters a virtual weightless state represented by computer graphics (CG). Visual information of a room and the degree-of-freedom of motion were varied to examine the influence of the conditions on such a simple task as movement through several connected modules, and the performance and the behavior of each subject were investigated.

Humans↗

Induction of three-dimensional assembly of human liver cells by simulated microgravity.

The establishment of long-term cultures of functional primary human liver cells (PHLC) is formidable. Developed at NASA, the Rotary Cell Culture System (RCCS) allows the creation of the unique microgravity environment of low shear force, high-mass transfer, and 3-dimensional cell culture of dissimilar cell types. The aim of our study was to establish long-term hepatocyte cultures in simulated microgravity. PHLC were harvested from human livers by collagenase perfusion and were cultured in RCCS. PHLC aggregates were readily formed and increased up to 1 cm long. The expansion of PHLC in bioreactors was further evaluated with microcarriers and biodegradable scaffolds. While microcarriers were not conducive to formation of spheroids, PHLC cultured with biodegradable scaffolds formed aggregates up to 3 cm long. Analyses of PHLC spheroids revealed tissue-like structures composed of hepatocytes, biliary epithelial cells, and/or progenitor liver cells that were arranged as bile duct-like structures along nascent vascular sprouts. Electron microscopy revealed groups of cohesive hepatocytes surrounded by complex stromal structures and reticulin fibers, bile canaliculi with multiple microvilli, and tight cellular junctions. Albumin mRNA was expressed throughout the 60-d culture. A simulated microgravity environment is conducive to maintaining long-term cultures of functional hepatocytes. This model system will assist in developing improved protocols for autologous hepatocyte transplantation, gene therapy, and liver assist devices, and facilitate studies of liver regeneration and cell-to-cell interactions that occur in vivo.

Albumins↗

Effect of simulated microgravity on aged pea seed vigour and related physiological properties.

We used simulated microgravity to treat aged pea seeds, in order to observe its effect on seed vigour and related physiological properties. The result shows that microgravity obviously promotes the germination rate, vitality index, germination index, and keeps the activity of protective enzyme, with conductivity and the content of MDA decreased. These effects gradually disappeared for a certain period after the microgravity administration.

Fabaceae↗

Increase in epinephrine-induced responsiveness during microgravity simulated by head-down bed rest in humans.

The epinephrine (Epi)-induced effects on the sympathetic nervous system (SNS) and metabolic functions were studied in men before and during a decrease in SNS activity achieved through simulated microgravity. Epi was infused at 3 graded rates (0.01, 0.02, and 0. 03 microg. kg(-1). min(-1) for 40 min each) before and on the fifth day of head-down bed rest (HDBR). The effects of Epi on the SNS (assessed by plasma norepinephrine levels and spectral analysis of systolic blood pressure and heart rate variability), on plasma levels of glycerol, nonesterified fatty acids (NEFA), glucose and insulin, and on energy expenditure were evaluated. HDBR decreased urinary norepinephrine excretion (28.1 +/- 4.2 vs. 51.5 +/- 9.1 microg/24 h) and spectral variability of systolic blood pressure in the midfrequency range (16.3 +/- 1.9 vs. 24.5 +/- 0.9 normalized units). Epi increased norepinephrine plasma levels (P < 0.01) and spectral variability of systolic blood pressure (P < 0.009) during, but not before, HDBR. No modification of Epi-induced changes in heart rate and systolic and diastolic blood pressures were observed during HDBR. Epi increased plasma glucose, insulin, and NEFA levels before and during HDBR. During HDBR, the Epi-induced increase in plasma glycerol and lactate levels was more pronounced than before HDBR (P < 0.005 and P < 0.001, respectively). Epi-induced energy expenditure was higher during HDBR (P < 0.02). Our data suggest that the increased effects of Epi during simulated microgravity could be related to both the increased SNS response to Epi infusion and/or to the beta-adrenergic receptor sensitization of end organs, particularly in adipose tissue and skeletal muscle.

Adrenergic alpha-Agonists↗