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PubMed1995

Increased beta-adrenergic responsiveness induced by 14 days exposure to simulated microgravity.

Increased sensitivity of end-organ responses to neuroendocrine stimuli as a result of prolonged exposure to the relative inactivity of microgravity has recently been hypothesized. This notion is based on the inverse relationship between circulating norepinephrine and beta-adrenoreceptor sensitivity. Beta-adrenoreceptor activity is reduced in individuals who have elevated plasma norepinephrine as as a result of regular exposure to upright posture and physical exercise. In contrast, adrenoreceptor hypersensitivity has been reported in patients with dysautonomias in which circulating catecholamines are absent or reduced. Taken together, these studies and the observation that circulating plasma norepinephrine has been reduced during spaceflight and in groundbased simulations of microgravity prompt the suggestion that adrenoreceptor hypersensitivity may be a consequence of the adaptation to spaceflight. We conducted an experiment designed to measure cardiovascular responses to adrenoreceptor agonists in human subjects before and after prolonged exposure to 6 degrees head-down tilt (HDT) to test the hypothesis that adaptation to microgravity increases adrenoreceptor responsiveness, and that this adaptation is associated with reduced levels of circulating norepinephrine.

Adrenergic alpha-Agonists↗
PubMed2001

Dome formation and tubule morphogenesis by Xenopus kidney A6 cell cultures exposed to microgravity simulated with a 3D-clinostat and to hypergravity.

Confluent high-density cell cultures of A6 cells derived from adult male Xenopus kidney exhibit spontaneous dome-formation at 1 g. To determine whether this morphogenetic property is altered by gravity, we used a three-dimensional (3D) clinostat to subject the cells to simulated microgravity, and a centrifuge to subject them to hypergravity. We used the generation orbit control method as the new rotation control system of the 3D-clinostat, not the random method. The growth of A6 cells was significantly enhanced by hypergravity, but significantly reduced by simulated microgravity. Dome formation by A6 cells at high confluence was inhibited under simulated microgravity conditions, whereas hypergravity promoted dome formation and induced tubule morphogenesis, compared to the control at 1 g. These results indicated that changes in gravity influence the morphogenetic properties of A6 cells, such as dome formation and tubule morphogenesis. When dome formation by A6 cells at high confluence was induced spontaneously in the control 1 g culture, the gene expression of the HGF family of pleiotropic factors, such as HGF-like protein (HLP) and growth factor-Livertine (GF-l.ivertine), an epithelial serine protease of channel activating protease 1 (CAP1), and Na+, K+-adenosine triphosphatase (ATPase), increased. Simulated microgravity increased the gene expression of activin A and reduced the gene expression of HLP, GF-Livertine, CAP1, and Na+, K+-ATPase. Hypergravity, on the other hand, decreased the gene expression of activin A and increased the gene expression of HLP, GF-Livertine, CAP1, and Na+, K+-ATPase. These results suggest that the effects of gravitational changes on expression of the HGF family member gene, CAP1, and Na+, K+-ATPase gene may be important for the cell growth, tubule morphogenesis, and dome formation of A6 cells in altered

Animals↗
PubMedDate not supplied

[The behavior of titin and the proteins of its family from skeletal muscles of ground squirrel (Citellus undulatus) during hibernation and rats under conditions of simulated microgravity].

By the use of SDS PAGE, the behavior of titin and MyBP-C in fast (m. psoas) as well as titin and MyBP-X in slow (m. soleus) muscles of ground squirrels (Citellus undulatus) during hibernation was compared with the behavior of titin and MyBP-X in rat m. soleus under conditions of simulated microgravity. A decrease in the amount of titin 1 and MyBP-C relative to that of myosin heavy chains by approximately 30% and approximately 40%, correspondingly, in muscles of hibernating and arousing ground squirrels was revealed in comparison with active animals. No differences in the relative amount of MyBP-X in m. soleus of hibernating, arousing and active ground squirrels were found. Under conditions of simulated microgravity, a decrease in the amount of titin 1 by approximately 2 times and MyBP-X by approximately1.5 times relative to that of myosin heavy chains in rat m. soleus was observed. By the method of SDS PAGE modified by us, an almost twofold decrease in the amount of short isovariants of the titin N2A isoform relative to that of myosin heavy chains was shown in muscles of hibernating and arousing ground squirrels, whereas no changes were found in the amount of long titin isovariants. The conditions of simulated microgravity resulted in a twofold decrease in the relative amount of both short and long titin isovariants in rat m. soleus. The results indicate that hibernating ground squirrels have an evolutionarily determined adaptive mechanism of selective degradation of fast muscle fibers and preservation or increase of slow fibers, as the most economic and energetically advantageous, with proteins typical of them. The microgravitation of nonhibernating animals (rats) leads to a non-selective degradation of MyBP-X and titin isovariants, which contributes to considerable atrophy of soleus fibers.

Adaptation, Physiological↗
PubMedDate not supplied

Taxane recovery from cells of Taxus in micro- and hypergravity.

Cell suspension cultures of Taxus cuspidata produce taxanes that are released from the outer surface of cells into the culture medium as free and bound alkaloids. Paclitaxel (Taxol (TM)), is an anti-cancer drug in short supply. It has a taxane ring derived from baccatin III and a C-13 phenylisoserine side-chain. This drug is produced over a wide range of gravitational forces. Monoclonal and polyclonal antibodies to paclitaxel, baccatin III, and the C-13 phenylisoserine side chain were combined in multiple-labeling studies to localize taxanes and paclitaxel on cell surfaces or on particles released into the culture medium. Bioreactor vessel design altered the composition of taxanes recovered from cells in simulated microgravity. At 10(-2) and 2x10(-4)g, taxane recovery was reduced but biomass growth and percent paclitaxel was significantly increased. At 1 to 24g, growth was reduced with a significant recovery of total taxanes with low percent paclitaxel. Bound paclitaxel was also localized in endonuclease-rich fragmenting nuclei of individual apoptotic cells. A model is presented comprising TCH (touch) genes encoding enzymes that modify taxane-bearing xylan residues in cell walls, the calcium-sensing of gravitational forces by the cytoplasm, and the predisposition of nuclei to apoptosis. This integrates the adaptive physiological and biochemical responses of drug-producing genomes with gravitational forces.

Apoptosis↗
PubMed2004

Formation and structure of transplantable tissue constructs generated in simulated microgravity from Sertoli cells and neuron precursors.

Cell transplantation therapy for Parkinson's disease (PD) has received much attention as a potential treatment protocol for this neurodegenerative condition. Although there have been promising successes with this approach, it remains problematic, especially regarding the inability to provide immediate trophic support to the newly grafted cells and the inability to prevent acute and/or long-term graft rejection by the host. To address these issues of cell graftability, we have created a novel tissue construct from isolated rat Sertoli cells (SC) and the NTerra-2 immortalized human neuron precursor cell line (NT2) utilizing NASA-developed simulated microgravity technology. The two cell types were cocultured at a 1:4 (SC/NT2) ratio in the High Aspect Rotating Vessel (HARV) biochamber for 3 days, after which a disc-shaped aggregate (1-4 mm diameter) was formed. Sertoli neuron aggregated cells (SNAC) were collected by gravity sedimentation and processed either for light and electron microscopy or for fluorescent immunocytochemistry. Intra-SNAC clusters of SC and NT2 cells were identified by anti-human mitochondrial protein (huMT--specific for NT2 cells) and cholera toxin subunit B (CTb--specific for SC). There was little evidence of cell death throughout the aggregate and the absence of central necrosis, as might be expected in such a large aggregate in vitro. Ultrastructurally, SC did not express junctional modifications with NT2 cells nor with adjacent SC as is typical of SC in vivo and, in some protocols, in vitro. NT2 cells, however, showed distinct intercellular junction-like densities with adjacent NT2 cells, often defining canaliculi-like channels between the microvillus borders of the cells. The results show that the use of simulated microgravity coculture provides a culture environment suitable for the formation of a unique and viable Sertoli-NT2 (i.e., SNAC) tissue construct displaying intra-aggregate cellular organization. The structural integration of SC with NT2 cells provides a novel transplantable tissue source, which can be tested to determine if SC will suppress rejection of the grafted NT2 cells and provide for their short- and long-term trophic support in situ in the treatment of experimental PD.

Animals↗
PubMed1999

Gravisensing, apoptosis, and drug recovery in Taxus cell suspensions.

Haploid and diploid cell suspensions of Taxus spp. were examined for their adaptive plasticity in response to simulated microgravity, unit gravity, and hypergravity. Cell suspensions produced the taxane, paclitaxel, (TAXOL (R)), which is useful for the treatment of various cancers. Amyloplasts contributed to taxane ring biosynthesis and to drug release at the cell wall. Drug-producing cells reacted as gravisensing osmotic tensiometers. In stressed cells, amyloplasts docked and fused in clusters to sites on the plasmalemma before taxane discharge into the culture medium. In simulated microgravity and compared to all other treatments, taxane production was reduced nearly 100-fold. The percent paclitaxel of total taxanes remained 3-to 6-fold greater, and biomass doubled. When p53-independent programmed cell death was induced, taxanes were released into the culture medium as free molecules (soluble and insoluble) or bound to membranes, nuclear fragments, xylan residues, and other particulate materials. Unit gravity and especially hypergravity promoted xylogenesis and significant drug overproduction. A model relating families of >touch = (TCH), taxane early response (TER), nuclear cycling, and apoptosis-regulating genes to gravisensing, cell wall modifications, and to taxane recovery accounted for most but not all of the observations.

Apoptosis↗
PubMed1998

Intracranial pressure dynamics during simulated microgravity using a new noninvasive ultrasonic technique.

It is believed that intracranial pressure (ICP) may be elevated in microgravity because a fluid shift toward the head occurs due to loss of gravitational blood pressures. Elevated ICP may contribute to space adaptation syndrome, because as widely observed in clinical settings, elevated ICP causes headache, nausea, and projectile vomiting, which are similar to symptoms of space adaptation syndrome. However, the hypothesis that ICP is altered in microgravity is difficult to test because of the invasiveness of currently-available techniques. We have developed a new ultrasonic technique, which allows us to record ICP waveforms noninvasively. The present study was designed to understand postural effects on ICP and assess the feasibility of our new device in future flight experiments.

Adult↗
PubMed1992

Microcomputer-based monitoring of cardiovascular functions in simulated microgravity.

A microcomputer-based system for non-invasive monitoring of cardiovascular system in simulated microgravity is described. The system evaluates automatically, accurately and interactively heart beat intervals, beat-to-beat non-invasive finger arterial blood pressure (systolic, diastolic, mean and pulse pressure) using a Finapres device and beat-to-beat changes of thoracic blood volume using impedance changes. In addition, beat-to-beat evaluation of cardiac mechanical function including left ventricular ejection time, diastolic time, systolic time intervals, left ventricular ejection fraction estimate and several other contractility parameters, left ventricular volume, stroke volume and cardiac output estimates are performed with high degree of automaticity.

Blood Pressure↗
PubMed1997

[Three dimensional (3D-) clinostat and its operational characteristics].

Three dimensional clinostat has been developed for simulation of microgravity on ground. It has applied in many disciplines in gravitational biology. Outline of operational principle is described together with its mechanical design. Rotation around two independent axes makes direction of gravity vector to scan whole steric angle. Magnitude and direction of rotational angular velocity is selected randomly at a certain interval of time to avoid singularity in sweep trajectory of gravity vector. Methods for validation of the operation are presented to test randomness of motion and cancellation of gravity by clino-rotation. Concerns discussed are vibration originated in motor and pseudo-weak magnetic field generated on clinostat. Fluid flow induced by clino-rotation is pointed as another problem to be taken into account.

Equipment Design↗
PubMed2000

Selected contribution: PKC activation inhibits Ca(2+) signaling in tracheal epithelial cells kept in simulated microgravity.

Microgravity has been shown to alter protein kinase C (PKC) activity; therefore, we investigated whether microgravity influences mechanically stimulated Ca(2+) signaling and ATP-induced Ca(2+) oscillations, both of which are modulated by PKC. Rabbit tracheal epithelial outgrowth cultures or suspended epithelial sheets were rotated in bioreactors to simulate microgravity. Mechanical stimulation of a single cell increased the cytosolic Ca(2+) concentration in 35-55 cells of both outgrowth cultures and epithelial sheets kept at unit gravity (G) or in simulated microgravity (smicroG). In outgrowth cultures, 12-O-tetradecanoylphorbol-13-acetate (TPA; 80 nM), a PKC activator, restricted Ca(2+) "waves" to about 10 cells in unit G and to significantly fewer cells in smicroG. TPA only slightly reduced the spread of Ca(2+) waves in epithelial sheets kept in smicroG but did not inhibit Ca(2+) waves of sheets kept in unit G. In both cell preparations from both conditions, TPA inhibited ATP-induced Ca(2+) oscillations; however, the effect was more pronounced in cells kept in smicroG. These results suggest that PKC activation is more robust in cells subjected to smicroG.

Adenosine Triphosphate↗
PubMed1994

Kinetics of stipe gravitropism in the mushroom fungus Coprinus cinereus under the conditions of microgravity simulation provided by clinostat treatment.

The extent of the gravitropic response, measured as the angle of the stipe apex at maximum curvature, was dependent upon the gravitational exposure time. The reaction time did not depend on exposure time, and interruption of gravitational exposure by a period of clinostating allowed the gravitational stimulus to decay. It is concluded that the gravitropic impulse is an 'all-or-nothing' signal in Coprinus cinereus, that perception and response probably occur in the same tissue regions, and that sustained exposure to the unidirectional gravity vector is necessary for the normal gravitropic response. The presentation time was found to be 7 min. Immediately after reaching curvature, stipes placed on the clinostat after various gravity exposure times 'relaxed' by 5 degrees. This relaxation suggests that gravitropic bending is a two-stage process with an initial, reversible, phase of plastic bending followed by a 'fixation' phase providing the gravitropic stimulus has been maintained.

Coprinus↗
PubMed1994

Acute effects of simulated microgravity on heart rate variability.

Microgravity is known to stress the heart and blood vessels and to perturb the normal neural regulation of the cardiovascular system. In an effort to gain greater insight into the adjustments of neural control of the cardiovascular system in space, we have used a model of simulated microgravity, the -6 degrees head-down tilt body position (HDT). As a common method for the description of the functional state of the cardiovascular system the power spectral analysis of the heart rate can be used. The following working hypotheses were postulated: 1. HDT causes an acute activation of the parasympathetic nerve traffic to the heart, which is detectable by a reduction in heart rate and by the spectral power distribution of the heart rate variability. 2. HDT induces changes in the autonomic nervous response to upright posture which can be detected after a 45-minute period of head-down tilt.

Adult↗
PubMed2002

[Stomatological studies on board of an "airspace" station].

The status of the oral organs was evaluated in normal subjects exposed for a long (30-262 days) time to conditions simulating an airspace flight in an on-land experimental complex (Sphinx-99 experiment). Stomatological studies were carried out in 15 volunteers aged 28-48 years. The initial (before "flight") and post-flight examination included clinical examination of the maxillofacial area and oral organs, evaluation of the intensity of caries by the CDL index, evaluation of the periodontal tissue status by the PMA and SPITN indexes. Based on the data, we singled out periods of adaptation characteristic of the oral organs under conditions of long enclosure. The results allow prediction of the course of diseases of the oral organs under extreme conditions and recommend methods of purposeful prevention in certain terms.

Adaptation, Physiological↗
PubMedDate not supplied

[Adaptive responses of the secretory cardiomyocytes of the right atrium during simulation of the long-term and repeated gravity by antiorthostatic suspension of rats].

Reactive changes in right atrium cardiomyocytes during antiorthostatic tail suspension of rats commonly used to simulate low gravity have been studied by electron microscopy and morphometry. A 14 day suspension proved to increase contractile and secreting activities of cardiomyocytes. At the same time, signs of depleted activity are observed in some cells. Elongation of the experiment to 30 days leads to development of adaptive compensatory responses and increases their secreting capacity. A 30 day return to normal orthostatic position does not completely restores the structure and functioning of cardiomyocytes and leads to accumulation of internal secretion. A repeated 14 day suspension to a certain extent facilitates cardiomyocyte adaptation to altered conditions as compared to a single exposure; apparently, secretion release decreases while its production is activated.

Adaptation, Physiological↗
PubMed2004

Microgravity alters basal and insulin-mediated metabolic activity of normal and neoplastic cells.

In this paper we report the behaviour of normal vascular smooth muscle cells and transformed breast cancer cells under normal versus simulated microgravity conditions by comparing cell proliferation, Glucose transport, Methionine uptake and protein synthesis. Modeled microgravity profoundly affects cell growth (especially in normal cells) and Glucose or Methionine metabolism (although to different extent in the two cell lines). Since both cells own responsive insulin receptors, the comparison was extended to insulin-stimulated versus unstimulated conditions. We report that the detected metabolic changes were strongly enhanced when the cells were simultaneously stimulated with insulin and subjected to modeled microgravity stress. Such observations may have important returns for human health in space; they deserve further attention.

Amino Acids↗
PubMed2005

A novel bioartificial liver with culture of porcine hepatocyte aggregates under simulated microgravity.

An extracorporeal bioartificial liver device could provide vital support to patients suffering from acute liver failure. We designed a novel, customized bioreactor for use as a bioartificial liver (patent pending). The Innsbruck Bioartificial Liver (IBAL) contains aggregates of porcine hepatocytes grown under simulated microgravity. The culture vessel rotates around its longitudinal axis and is perfused by two independent circuits. The circuit responsible for exchange of plasma components with the patient consists of a dialysis tube winding spirally around the internal wall of the culture vessel. IBAL was evaluated in vitro. Viability tests showed sufficient viability of hepatocytes for up to 10 days. Cytologic examination of samples from the bioreactor showed liver cell aggregates. These were also examined by electron microscopy. A number of biochemical parameters were analyzed. In conclusion, cell culture is possible for at least 10 days in the IBAL system, organoid hepatocyte aggregates are formed and synthetic activity of the hepatocytes was demonstrated.

Animals↗
PubMed2004

[Pilot study of neonatal rat cardiac myocytes cultured for three-dimensional modeling in simulated microgravity].

OBJECTIVE: To study three-dimensional culturing methods of neonatal rat cardiac myocytes in simulated microgravity. METHODS: Neonatal rat primary cardiac myocytes were separated and seeded into polylactic acid scaffolds, stirred in spinner flasks for 24 hours, and then moved into rotary cell culture system for three-dimensional culture. The growth of cardiac myocytes was observed under inverted phase contrast microscope, scanning electron microscope and transmission electron microscope, and metabolic assay was assessed by MTT assay. RESULTS: Cardiac myocytes with sustained metabolic activity attached to the polylactic acid scaffolds, extended and confluenced. Pulsations of PLA-cardiac myocytes was found in some areas. CONCLUSION: The rotary cell culture system is suitable to develop neonatal rat cardiac myocytes culturing for three-dimensional modeling.

Animals↗
PubMed2006

[Disruption of microfilament cytoskeleton induced by simulated microgravity increases the activity of COL1A1 promoter].

It is well known that cytoskeleton system is the sensor of gravity in cells. Under microgravity condition, cytoskeleton is associated with the changes of cell shape, function, signaling and so on; but the relationship between cytoskeleton and gene expression is not fully understood. In present study, we discussed the effects of cell microfilament on the activity of collagen type I alpha 1 chain gene (COL1A1) promoter under microgravity simulated by clinostat and/or cytochalasin B as microfilament depolymerizer in the established EGFP-ROS cell line using the method of fluorescence semi-quantitative analysis and the fluorescent stain of microfilament. Compared with the normal control, the microfilament of ROS17/2.8 cell tended to disassemble, marginal distribution of fiber stress, and showed reducing stress fibers after spaceflight in Photon-M1 or clinorotation simulated microgravity, which suggested that microgravity destroyed the well-order cell cytoskeleton and induced a rearrangement. Treatment with suitable concentration of cytochalasin B in normal gravity induced disruption of microfilament, increased the activity of COL1A1 promoter and resulted in a dose-dependent increase of EGFP fluorescence. Therefore, a certain extent disruption of the microfilament system was associated with increased activity of the COL1A1 promoter. All above demonstrate that microfilament cytoskeleton system takes part in the regulation of COL1A1 promoter activity and plays an important role in the signaling of microgravity.

Actin Cytoskeleton↗