Effects Of Hypergravity Research Articles

Cell Wall Assembly and Intracellular Trafficking in Plant Cells Are Directly Affected by Changes in the Magnitude of Gravitational Acceleration

Plants are able to sense the magnitude and direction of gravity. This capacity is thought to reside in selected cell types within the plant body that are equipped with specialized organelles called statoliths. However, most plant cells do not possess statoliths, yet they respond to changes in gravitational acceleration. To understand the effect of gravity on the metabolism and cellular functioning of non-specialized plant cells, we investigated a rapidly growing plant cell devoid of known statoliths and without gravitropic behavior, the pollen tube. The effects of hyper-gravity and omnidirectional exposure to gravity on intracellular trafficking and on cell wall assembly were assessed in Camellia pollen tubes, a model system with highly reproducible growth behavior in vitro. Using an epi-fluorescence microscope mounted on the Large Diameter Centrifuge at the European Space Agency, we were able to demonstrate that vesicular trafficking is reduced under hyper-gravity conditions. Immuno-cytochemistry confirmed that both in hyper and omnidirectional gravity conditions, the characteristic spatial profiles of cellulose and callose distribution in the pollen tube wall were altered, in accordance with a dose-dependent effect on pollen tube diameter. Our findings suggest that in response to gravity induced stress, the pollen tube responds by modifying cell wall assembly to compensate for the altered mechanical load. The effect was reversible within few minutes demonstrating that the pollen tube is able to quickly adapt to changing stress conditions.

The relationship between brain cortical activity and brain oxygenation in the prefrontal cortex during hypergravity exposure

Artificial gravity has been proposed as a method to counteract the physiological deconditioning of long-duration spaceflight; however, the effects of hypergravity on the central nervous system has had little study. The study aims to investigate whether there is a relationship between prefrontal cortex brain activity and prefrontal cortex oxygenation during exposure to hypergravity. Twelve healthy participants were selected to undergo hypergravity exposure aboard a short-arm human centrifuge. Participants were exposed to hypergravity in the +Gz axis, starting from 0.6 +Gz for women, and 0.8 +Gz for men, and gradually increasing by 0.1 +Gz until the participant showed signs of syncope. Brain cortical activity was measured using electroencephalography (EEG) and localized to the prefrontal cortex using standard low-resolution brain electromagnetic tomography (LORETA). Prefrontal cortex oxygenation was measured using near-infrared spectroscopy (NIRS). A significant increase in prefrontal cortex activity (P < 0.05) was observed during hypergravity exposure compared with baseline. Prefrontal cortex oxygenation was significantly decreased during hypergravity exposure, with a decrease in oxyhemoglobin levels (P < 0.05) compared with baseline and an increase in deoxyhemoglobin levels (P < 0.05) with increasing +Gz level. No significant correlation was found between prefrontal cortex activity and oxy-/deoxyhemoglobin. It is concluded that the increase in prefrontal cortex activity observed during hypergravity was most likely not the result of increased +Gz values resulting in a decreased oxygenation produced through hypergravity exposure. No significant relationship between prefrontal cortex activity and oxygenation measured by NIRS concludes that brain activity during exposure to hypergravity may be difficult to measure using NIRS. Instead, the increase in prefrontal cortex activity might be attributable to psychological stress, which could pose a problem for the use of a short-arm human centrifuge as a countermeasure.

The Impact of Altered Gravity and Vibration on Endothelial Cells During a Parabolic Flight

Background: Endothelial cells (EC) cultured under altered gravity conditions show a cytoskeletal disorganization and differential gene expression (short-term effects), as well as apoptosis in adherently growing EC or formation of tubular 3D structures (long-term effects). Methods: Investigating short-term effects of real microgravity, we exposed EC to parabolic flight maneuvers and analysed them on both protein and transcriptional level. The effects of hypergravity and vibration were studied separately. Results: Pan-actin and tubulin proteins were elevated by vibration and down-regulated by hypergravity. β-Actin was reduced by vibration. Moesin protein was reduced by both vibration and hypergravity, ezrin potein was strongly elevated under vibration. Gene expression of ACTB, CCND1, CDC6, CDKN1A, VEGFA, FLK-1, EZR, ITBG1, OPN, CASP3, CASP8, ANXA2, and BIRC5 was reduced under vibration. With the exception of CCNA2, CCND1, MSN, RDX, OPN, BIRC5, and ACTB all investigated genes were downregulated by hypergravity. After one parabola (P) CCNA2, CCND1, CDC6, CDKN1A, EZR, MSN, OPN, VEGFA, CASP3, CASP8, ANXA1, ANXA2, and BIRC5 were up-, while FLK1 was downregulated. EZR, MSN, OPN, ANXA2, and BIRC5 were upregulated after 31P. Conclusions: Genes of the cytoskeleton, angiogenesis, extracellular matrix, apoptosis, and cell cycle regulation were affected by parabolic flight maneuvers. We show that the microgravity stimulus is stronger than hypergravity/vibration.

Hypergravity and Multiple Reflections in Wave Propagation in the Aorta

Hypergravity and gravity changes encountered in, e.g., airplanes, rollercoasters, and spaceflight can result in headaches or loss of consciousness due to decreased cerebral blood flow. This paper describes the effect of hypergravity and gravity changes on the pressure in the aorta and the distension of its wall. The model presented consists of a pressure part caused by gravity and a part representing pressure waves propagating through the vessel. The total pressure is described by a one-dimensional formulation in the frequency domain. To accommodate for geometrical and material variations, the vessel is modeled as a series of sections in which multiple reflections can occur. Results are presented for constant and varying gravity in straight and tapered flexible vessels.

Mushy-layer dynamics in micro and hyper gravity

We describe the results of experiments on mushy layers grown from aqueous ammonium chloride solution in normal, micro, and hyper gravity environments. In the fully developed chimney state, the chimney plume dynamics differ strikingly when conditions change from micro to hyper gravity. In microgravity, we find fully arrested plume motion and suppressed convection. As gravity exceeds Earth conditions, we observe a host of phenomena, ranging from arched plumes that undergo forced Rayleigh-Taylor instabilities to in-phase multiple plume oscillatory behavior. For the same initial solute concentrations and fixed boundary cooling temperatures, we find that, in runs of over two hours, the averaged effects of microgravity and hypergravity result in suppressed growth of the mushy layers, a phenomenon caused by a net enhancement of convective heat and solute transport from the liquid to the mushy layers. These behaviors are placed in the context of the theory of convecting mushy layers as studied under normal laboratory conditions.

Behavioral responses to partial-gravity conditions in rats

The effects of microgravity or hypergravity on living organisms have been studied extensively; however, thus far no studies have addressed the effects of “partial-gravity”, that is, the low-gravity levels between the unit gravity (1G) on Earth and zero gravity (0G) in space. The purpose of the present study was to examine behavioral responses in rats under partial-gravity conditions. Rat behavior was monitored by video cameras during parabolic flights. The flight trajectory was customized in order to generate graded levels of partial gravity. Gravity-dependent behavior patterns were observed in rats. In the conditions of 0.4G through 0.2G, rats showed startle and crouching. Hindlimb stretching emerged at 0.15G and was more frequently observed toward 0.01G. Different thresholds may exist for emotional and balance/posture-related behaviors.

Effects of gravistimuli on osmoregulation in azuki bean epicotyls

The effects of hypergravity on growth and osmoregulation were examined in dark-grown azuki bean epicotyls. Elongation growth of epicotyls was promptly suppressed by hypergravity at 300g. On the contrary, the increase in fresh weight of epicotyls during incubation was not suppressed by hypergravity at 300g at least up to 6h. Also, the level of total osmotic solutes increased during epicotyl growth for 6h, which was not affected by hypergravity. These results suggest that azuki bean epicotyls are capable of maintaining osmoregulation even under 300g conditions for a short period. On the other hand, the increase in fresh weight of epicotyls was suppressed, in addition to suppression of elongation growth, when seedlings were treated with 300g for 24h. The increase in level of total osmotic solutes was also inhibited by 24h hypergravity treatment, which was accounted by the reduced levels of organic solutes, such as sugars and amino acids. Furthermore, the dry weight of seeds decreased during incubation for 24h, but the decrease was inhibited by hypergravity at 300g. Hypergravity treatment at 300g for 24h also increased the pH value of apoplastic solution in epicotyls. Taken together, these results suggest that the translocation of organic solutes from the seed to epicotyls is inhibited by prolonged hypergravity treatment, which may underlie the suppression of epicotyl growth, and that the breakdown of H+ gradient across the plasma membrane in epicotyl cells may be at least partly involved in the reduction of organic solute accumulation under hypergravity conditions.

The Effects of Hypergravity on Xenopus Embryo Growth and Cardiac Hypertrophy

All life on earth has developed and evolved in a unity gravity (1G) environment. Any deviation below or above 1G could affect animal development, a period when much change occurs and sensitivity is high. We imposed simulated hypergravity through centrifugation and analyzed the effects on the overall body length and cardiac growth of Xenopus laevis embryos. We predicted that increased contractile force would be required from the heart to adequately circulate blood, dispersing nutrients, and that this would inhibit organism growth and possibly induce a state of hypertrophy. Embryos reaching gastrulation stage were exposed to a 7G or 1G (control) field via centrifugation for 96 hours. We then recorded behavior, mortality and took body length measurements. We found no significant differences in behavior or mortality, however, body length was significantly reduced by an average of 6.8% in the 7G group. We then fixed, embedded, sectioned and stained embryos in order to investigate the dimensions of cardiac tissue and of the cardiac region of the body using image analysis software. We found the 7G group had a significantly reduced average body cross-sectional area (-18%) and yet a significantly larger ventricular cross-sectional area (+36%) when compared to the 1G group. The average ratio of ventricle cross-sectional area to average body cross-sectional area was significantly higher in the 7G group when compared to the 1G. From these data, we conclude that hypergravity has a significant inhibitory impact on the Xenopus laevis embryo growth and causes a significant increase in ventricle size.

Effect of Hypergravity and Phytohormones on Isoflavonoid Accumulation in Soybean (Glycine max. L.) Callus

The objective of this study was to explore the potential interaction between gravity and growth hormones on isoflavonoid accumulation. Soybean callus (Glycine max (L.) Merr. cv. ‘Acme’) was grown in the dark for 16 days at 22 °C in a growth medium supplemented with four different combinations of phytohormones and subjected to 4-g and 8-g forces simulated in a centrifuge and 1-g in an adjacent stationary control. Isoflavonoid aglycones and their glycoside concentrations (daidzein, genistein, daidzin, 6″-O-malonyl-7-O-glucosyl daidzein, genistin, 6″-O-malonyl-7-O-glucosyl genistein) were determined in the resulting tissues. Although gravity had no significant impact on callus growth, increasing gravity reduced isoflavonoid accumulation in three out of the four phytohormone-supplemented culture media. The ratio of the auxin naphthalene acetic acid (NAA) to the cytokinin benzylaminopurine (BAP) was found to have profound effect on both callus growth and isoflavonoid accumulation. The cytokinin BAP promoted callus tissue growth, but reduced callus isoflavonoid suggesting the isoflavonoid accumulation was not keeping pace with the cell growth in the elevated concentration of BAP. On the other hand, NAA had little or no effect on callus growth, but greatly enhanced isoflavonoid accumulation. Interactive effects of gravity and hormone on isoflavonoid accumulation were evident and its implication to the mechanism by which gravity exerts the effect on plant secondary metabolites is discussed.

A Hypergravity Environment Induced by Centrifugation Alters Plant Cell Proliferation and Growth in an Opposite Way to Microgravity

Seeds of Arabidopsis thaliana were exposed to hypergravity environments (2g and 6g) and germinated during centrifugation. Seedlings grew for 2 and 4 days before fixation. In all cases, comparisons were performed against an internal (subjected to rotational vibrations and other factors of the machine) and an external control at 1g. On seedlings grown in hypergravity the total length and the root length were measured. The cortical root meristematic cells were analyzed to investigate the alterations in cell proliferation, which were quantified by counting the number of cells per millimeter in the specific cell files, and cell growth, which were appraised through the rate of ribosome biogenesis, assessed by morphological and morphometrical parameters of the nucleolus. The expression of cyclin B1, a key regulator of entry in mitosis, was assessed by the use of a CYCB1:GUS genetic construction. The results showed significant differences in some of these parameters when comparing the 1g internal rotational control with the 1g external control, indicating that the machine by itself was a source of alterations. When the effect of hypergravity was isolated from other environmental factors, by comparing the experimental conditions with the rotational control, cell proliferation appeared depleted, cell growth was increased and there was an enhanced expression of cyclin B1. The functional meaning of these effects is that cell proliferation and cell growth, which are strictly associated functions under normal 1g ground conditions, are uncoupled under hypergravity. This uncoupling was also described by us in previous experiments as an effect of microgravity, but in an opposite way. Furthermore, root meristems appear thicker in hypergravity-treated than in control samples, which can be related to changes in the cell wall induced by altered gravity.

Hypergravity effects on myoblast proliferation and differentiation

This study aimed at the investigation of behavior of myoblasts in conditions of altered gravity. C2C12 cells underwent stimulations by different hypergravity intensities (2 h at 5 g, 10 g, and 20 g) in the Large Diameter Centrifuge of the European Space Agency (ESA), highlighting positive effects on both proliferation and differentiation.

Involvement of auxin dynamics in hypergravity-induced promotion of lignin-related gene expression in Arabidopsis inflorescence stems.

Recent studies have shown that hypergravity enhances lignification through up-regulation of the expression of lignin biosynthesis-related genes, although its hormonal signalling mechanism is unknown. The effects of hypergravity on auxin dynamics were examined using Arabidopsis plants that were transformed with the auxin reporter gene construct DR5::GUS. Hypergravity treatment at 300 g significantly increased β-glucuronidase activity in inflorescence stems of DR5::GUS plants, indicating that endogenous auxin accumulation was enhanced by hypergravity treatment. The hypergravity-related increased expression levels of both DR5::GUS and lignin biosynthesis-related genes in inflorescence stems were suppressed after disbudding, indicating that the increased expression of lignin biosynthesis-related genes is dependent on an increase in auxin influx from the shoot apex.

Inhibition of Gene Expression inEscherichia coliunder Hypergravity

We investigated the effects of hypergravity on gene expression in Escherichia coli. Analysis of gene expression using green fluorescence protein reporter showed downregulation at 50,000 g, as found for OmpW, which was one of the gravity-induced proteins. Real time-PCR analysis showed normal transcription even at 50,000 g. These results suggest that hypergravity inhibited the translation process in gene expression.

Differential Gene Regulation under Altered Gravity Conditions in Follicular Thyroid Cancer Cells: Relationship between the Extracellular Matrix and the Cytoskeleton

Extracellular matrix proteins, adhesion molecules, and cytoskeletal proteins form a dynamic network interacting with signalling molecules as an adaptive response to altered gravity. An important issue is the exact differentiation between real microgravity responses of the cells or cellular reactions to hypergravity and/or vibrations. To determine the effects of real microgravity on human cells, we used four DLR parabolic flight campaigns and focused on the effects of short-term microgravity (22 s), hypergravity (1.8 g), and vibrations on ML-1 thyroid cancer cells. No signs of apoptosis or necrosis were detectable. Gene array analysis revealed 2430 significantly changed transcripts. After 22 s microgravity, the F-actin and cytokeratin cytoskeleton was altered, and ACTB and KRT80 mRNAs were significantly upregulated after the first and thirty-first parabolas. The COL4A5 mRNA was downregulated under microgravity, whereas OPN and FN were significantly upregulated. Hypergravity and vibrations did not change ACTB, KRT-80 or COL4A5 mRNA. MTSS1 and LIMA1 mRNAs were downregulated/slightly upregulated under microgravity, upregulated in hypergravity and unchanged by vibrations. These data indicate that the graviresponse of ML-1 cells occurred very early, within the first few seconds. Downregulated MTSS1 and upregulated LIMA1 may be an adaptive mechanism of human cells for stabilizing the cytoskeleton under microgravity conditions.

Gravity Sensing byEscherichia coli

We investigated the growth and protein profile of Escherichia coli under various gravity strengths to determine the effects of hypergravity on biochemical reactions. E. coli grows at less than 7,500 g without inhibition. Hypergravity induced OmpW and Antigen 43. Changes in gravity strength altered the expression levels of these proteins. This suggests that hypergravity regulates gene expression in bacteria.

Hypergravity suppresses bone resorption in ovariectomized rats

The effects of gravity on bone metabolism are unclear, and little has been reported about the effects of hypergravity on the mature skeleton. Since low gravity has been shown to decrease bone volume, we hypothesized that hypergravity increases bone volume. To clarify this hypothesis, adult female rats were ovariectomized and exposed to hypergravity (2.9G) using a centrifugation system. The rats were killed 28days after the start of loading, and the distal femoral metaphysis of the rats was studied. Bone architecture was assessed by micro-computed tomography (micro-CT) and bone mineral density was measured using peripheral quantitative CT (pQCT). Hypergravity increased the trabecular bone volume of ovariectomized rats. Histomorphometric analyses revealed that hypergravity suppressed both bone formation and resorption and increased bone volume in ovariectomized rats.Further, the cell morphology, activity, proliferation, and differentiation of osteoclasts and osteoblasts exposed to hypergravity were evaluated in vitro. Hypergravity inhibited actin ring formation in mature osteoclasts, which suggested that the osteoclast activity was suppressed. However, hypergravity had no effect on osteoblasts. These results suggest that hypergravity can stimulate an increase in bone volume by suppressing bone resorption in ovariectomized rats.

Effects of Hypergravity on Formation of Microporous Films by Dissipative Processes

Microporous polymer films can be prepared by casting a solution of hydrophobic polymer and amphiphilic polymer on a solid substrate using condensed water droplet arrays as templates. Bacause of the temperature nonuniformity of the solution surface owing to thermal convections, water condenses heterogeneously. The heterogeneous condensation of water droplets induces the formation of irregular-size and -arrangement microporous film. To evaluate the effect of gravity on microporous film formation, we have successfully prepared the test chamber for hypergravity experiments. From the hypergravity experiment, the thermal convection during solvent evaporation was forced to be enhanced under hypergravity. As a result, the microporous structure was not observed under hypergravity. These results provided insight on water-assisted microporous film formation. Furthermore, the results strongly suggested that the microgravity enviroment is suitable for uniform and well-organized microporous film formation.

Gravity Scaling Parameter for Pool Boiling Heat Transfer

Although the effects of microgravity, earth gravity, and hypergravity (&gt;1.5 g) on pool boiling heat flux have been studied previously, pool boiling heat flux data over a continuous range of gravity levels (0–1.7 g) was unavailable until recently. The current work uses the results of a variable gravity, subcooled pool boiling experiment to develop a gravity scaling parameter for n-perfluorohexane/FC-72 in the buoyancy-dominated boiling regime (Lh/Lc&gt;2.1). The heat flux prediction was then validated using heat flux data at different subcoolings and dissolved gas concentrations. The scaling parameter can be used as a tool to predict boiling heat flux at any gravity level in the buoyancy dominated regime if the data under similar experimental conditions are available at any other gravity level.

Computer Modeling Describes Gravity-Related Adaptation in Cell Cultures

Questions about the changes of biological systems in response to hostile environmental factors are important but not easy to answer. Often, the traditional description with differential equations is difficult due to the overwhelming complexity of the living systems. Another way to describe complex systems is by simulating them with phenomenological models such as the well-known evolutionary agent-based model (EABM). Here we developed an EABM to simulate cell colonies as a multi-agent system that adapts to hyper-gravity in starvation conditions. In the model, the cell's heritable characteristics are generated and transferred randomly to offspring cells. After a qualitative validation of the model at normal gravity, we simulate cellular growth in hyper-gravity conditions. The obtained data are consistent with previously confirmed theoretical and experimental findings for bacterial behavior in environmental changes, including the experimental data from the microgravity Atlantis and the Hypergravity 3000 experiments. Our results demonstrate that it is possible to utilize an EABM with realistic qualitative description to examine the effects of hypergravity and starvation on complex cellular entities.

The Mechanisms for Platelet Hyperactivity Induced by Hypergravity.

Abstract 5059Many gravity change-related thrombotic and hemorrhagic diseases or fatalities have been reported to occur in human beings, such as crewmen in space, roller coaster riders, and aircrew subjected to high-G training. Because platelets, as anucleated blood cell, are the primary players in both thrombosis and hemostasis, function change of platelets in altered gravity conditions might offer some insights into the mechanisms leading to these gravity change-related diseases. We have demonstrated that the platelet functions are inhibited in microgravity environments, and activated under high-G conditions, which reveals a novel mechanism for gravity change-related hemorrhagic and thrombotic diseases. The mechanisms, however, remain to be further investigated. Here, firstly we investigated the effect of hypergravity (acceleration centrifugation) on platelet functions. Platelet rich plasma (PRP) or washed platelets were exposed to hypergravity at 8 G for different time points (2 minutes, 10 minutes, 15 minutes, 20 minutes), and then platelet aggregation was measured at each time point. No platelet aggregation was induced by 8 G alone, whereas ristocetin or collagen-induced platelet aggregation was time-dependently increased by hypergravity stimulus. The effect was maximal at 15 minutes after hypergravity treatment. Spreading assay shows that the number of platelet adhesion to immobilized fibrinogen and the mean platelet area spreading on von Willbrand factor (VWF) matrix were obviously increased by hypergravity. To further explore the effect of hypergravity on platelet activity state, the markers of platelet activations (integrin αaIIbβ3 activation, P-selectin surface expression) were assessed by flow cytometry. αaIIbβ3 was activated, in part, in 8 G-exposed platelets as detected by PAC-1 binding, but no significant difference was observed in P-selectin surface expression compared with 1 G controls. Next, we investigated the effects of hypergravity stimulus on actin distribution and intracellular Ca2+ ([Ca2+]i) in isolated human platelets. Hypergravity at 8 G induced actin cytoskeleton reorganization in platelets, and the release of Ca2+ from internal stores, but not Ca2+ influx via mechanosensitive Ca2+ channels in the plasma membrane. Pretreatment of platelets with intercellular Ca2+ chelator BAPTA/AM had no effect on hypergravity-induced changes in cytoskeleton, but significantly reduced platelet aggregation induced by hypergravity/ristocetin. In addition, using two different platelet activation inhibitors acetylsalicylic acid (aspirin) and tirofiban, which are widely used as antithrombotic drugs, we assessed the effects of the drugs on hypergravity-induced platelet activation in mice. Treatment of mice with aspirin or tirofiban significantly reduced hypergravity-induced P-selectin expression on platelet surface, prolonged tail bleeding time in vivo and reduced death rate in hypergravity-treated mice. Thus, our results indicate that hypergravity leads to human platelet hyperactivity, but fails to incur essential platelet activation events. Actin cytoskeleton reorganization and [Ca2+]i play important roles in regulation of platelet functions under hypergravity condition. The reorganization of actin cytoskeleton may be upstream or independent of [Ca2+]i, suggesting that hypergravity would affect various intercellular signals leading to platelet hyperactivity. In addition, antithrombotic agents such as aspirin and tirofiban have the therapeutic potential for hypergravity-induced thrombotic diseases. DisclosuresNo relevant conflicts of interest to declare.