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[Effects of acute hypobaric hypoxia on gastric emptying and intestinal propulsion: experiment with rats].

OBJECTIVE: To investigate the impact of acute hypobaric hypoxia on the gastrointestinal motility. METHODS: Eighty Wistar rats were randomly divided into 4 equal groups to be fed with (99)Tc(m)-labeled test food: ground level control group, put in the hypobaric chamber for 30 minutes; 3000 m simulated altitude group, exposed to the environment of simulated altitude of 3000 m for 30 minutes; 5000 m simulated altitude group, exposed to the environment of simulated altitude of 5000 m for 30 minutes; and mosapride + 5000 m simulated altitude group, fed with mosapride 2 mg/kg by perfusing stomach and fed with isotope-labeled test food 30 minutes later, and then exposed to 5000 m simulated altitude for 30 minutes. By the end of experiment the rats were killed, their stomachs were taken out to calculate the gastric emptying rate. Their intestine from pylorus to ileocecum was taken out to measure the intestinal propulsion function by using charcoal particle method. At the beginning and at the end of experiment abdominal arterial blood samples were collected to detect the plasma motilin and nitric oxide (NO) concentrations. RESULTS: The gastric emptying rate of the 5000 m simulated altitude group was 41% +/- 10%, significantly lower than that of the ground level group (62% +/- 12%, P < 0.01), and the charcoal transit rate of the 5000 m simulated altitude group was 37% +/- 8%, significantly lower than that of the ground level group (61% +/- 13%, P < 0.01). The gastric emptying rate and intestine propulsion rate of the 3000 m simulated altitude group were not significantly different from those of the ground level group. The gastric emptying rate of the mosapride + 5000 m simulated altitude group was 55% +/- 12%, significantly higher than that of the 5000 m simulated altitude group (P < 0.05), however, the intestine propulsion rate of the mosapride + 5000 m simulated altitude group was not significantly different from that of the 5000 m simulated altitude group (P > 0.05). The plasma motilin level of the 5000 m simulated altitude group was 88 pg/ml +/- 19 pg/ml, significantly lower than that of the ground level group (123 pg/ml +/- 28 pg/ml, P < 0.01), in contrast, the plasma NO level of the 5000 m simulated altitude group was 106 micromol/L +/- 24 micromol/L, significantly higher than that of the ground level group (80 micromol/L +/- 18 micromol/L, P < 0.01). CONCLUSION: Acute exposure to hypobaric hypoxia at the height of 5000 m inhibits the gastric emptying and intestinal propulsion. Mosapride may alleviate the inhibitory effect of hypobaric hypoxia on gastric emptying. Decrease of plasma motilin and elevation of NO level may be the main mechanism of inhibition of gastrointestinal motility by hypobaric hypoxia.

Animals↗

Muscles in microgravity: from fibres to human motion.

In simulated or actual microgravity, human and animal postural muscles undergo substantial atrophy: after about 270 days, the muscle mass attains a constant value of about 70% of the initial one. Most animal studies reported preferential atrophy of slow twitch fibres whose mechanical properties change towards the fast type. However, in humans, at the end of a 42-days bed rest study, a similar atrophy of slow and fast fibres was observed. After microgravity, the maximal force of several muscle groups showed a substantial decrease (6-25% of pre-flight values). The maximal power during very short "explosive" efforts of 0.25-0.30s showed an even greater fall, being reduced to 65% after 1 month and to 45% (of pre-flight values) after 6 months. The maximal power developed during 6-7s "all-out" bouts on an isokinetic cycloergometer was reduced to a lesser extent, attaining about 75% of pre-flight values, regardless of the flight duration. In these same subjects, the muscle mass of the lower limbs declined by only 9-13%. Thus, a substantial fraction of the observed decreases of maximal power is probably due to a deterioration of the motor co-ordination brought about by the absence of gravity. To prevent this substantial decay of maximal absolute power, we propose that explosive exercise be added to the daily in-flight training schedule. We also describe a system aimed at reducing cardiovascular deconditioning wherein gravity is simulated by the centrifugal acceleration generated by the motion of two counter rotating bicycles ridden by the astronauts on the inner wall of a cylindrical space module. Finally, cycling on circular or elliptical tracks may be useful to reduce cardiovascular deconditioning in permanently manned lunar bases. Indeed, on the curved parts of the path, a cyclist generates an outward acceleration vector (ac). To counterbalance ac, the cyclist must lean inwards, so that the vectorial sum of ac plus the lunar gravity tends to the acceleration of gravity prevailing on Earth.

Animals↗

Prevention of human deconditioning during prolonged immersion in water.

A 56-day immersion experiment in which two subjects participated was carried out. During the experiment the preventive effect of periodic acceleration combined with exercise and water-salt intake was assessed. Simulating an increased gravitational field, exposure to acceleration increased the static component of the load upon the musculo-skeletal system, increased the gradient of the blood hydrostatic pressure, activated mechanisms responsible for the venous return to the heart, stimulated systems regulating antidiuretic and antisodiumdiuretic reflexes. Involvement of these mechanisms restored haemodynamic parameters, fluid-electrolyte balance and blood coagulability. The prophylactic effect of acceleration was enhanced if combined with exercise and supplemented water-salt intake.

Acceleration↗

Response to 45 degrees head-down tilt as measured by organ weight/body weight ratios and spiral computed tomography.

BACKGROUND: Exposure to microgravity or simulated microgravity causes significant shifts in body fluids which may initiate physiological adaptations to the microgravity stressor. It is imperative to understand the physiological adaptations to microgravity in order to develop appropriate countermeasures to the deleterious aspects (i.e., muscle and bone wasting) of long-term spaceflights. HYPOTHESIS: The significant shifts in body fluids by 45 degrees head-down tilt can be measured by changes in organ weight/body weight (OW/BW) ratios and non-invasively by spiral computed tomography. METHODS: In a previous study (14), rats were weighed and exposed to either 45 degrees head-down tilt (45HDT) or a prone control position for one of the following experimental times: 0.5 h, 1 h, 2 h, 4 h, 8 h, or 24 h. A radioactive tracer was injected intramuscularly immediately prior to the start of the experimental time periods. At the end of the experiment, the major organs were harvested, weighed, and measured for gamma radiation levels. We used the organ weights from this previous study to calculate OW/BW ratios for the present study. Additionally, in the present study, rats in the 14-d experimental groups were weighed, lightly anesthetized to facilitate placement in the 45HDT position, and placed in a specially designed 45HDT cage (45HDT group) or left unrestrained in the cages (control group). At the end of the 14-d experimental time period, the rats were anesthetized and their lung densities measured with spiral computed tomography. RESULTS: The OW/BW ratios for the liver, kidneys, and spleen of 24 h 45HDT rats were significantly lower (p<0.05) than control values while at 1 h the 45HDT rats had a higher kidney OW/BW ratio. Lung density from the 14-d 45HDT rats was 24.4% greater than control rats' values. CONCLUSIONS: The physiological change due to the 45HDT position to simulate microgravity begins as early as 1 h, and the kidney appears to be the first organ affected. Spiral computed tomography may offer a viable method of non-invasively measuring organ densities in the 45HDT model. The OW/BW data generated in the present study does not correlate with the changes in radioactive tracer distribution data from our previous study.

Adaptation, Physiological↗

Reactive oxygen species and antioxidant enzymes activity of Anabaena sp. PCC 7120 (Cyanobacterium) under simulated microgravity.

It was found that reactive oxygen species in Anabaena cells increased under simulated microgravity provided by clinostat. Activities of intracellular antioxidant enzymes, such as superoxide dismutase, catalase were higher than those in the controlled samples during the 7 days' experiment. However, the contents of glutathione [correction of gluathione], an intracellular antioxidant, decreased in comparison with the controlled samples. The results suggested that microgravity provided by clinostat might break the oxidative/antioxidative balance. It indicated a protective mechanism in algal cells, that the total antioxidant system activity increased, which might play an important role for algal cells to adapt the environmental stress of microgravity.

Anabaena↗

Effects of gravity on early development.

The development of embryonic and larval stages of the South African Toad Xenopus laevis D, was investigated in hyper-g up to 5 g (centrifuge), in simulated 0 g (fast-rotating clinostat), in alternating low g, hyper-g (parabolic flights) and in microgravity (Spacelab missions D1, D-2). The selected developmental stages are assumed to be very sensitive to environmental stimuli. The results showed that the developmental reaction processes run normal also in environments different to 1 g and that aberrations in behavior and morphology normalize after return to 1 g. Development, differentiation, and morphology of the gravity perceiving parts of the vestibular system (macula-organs) had not been affected by exposure to different g-levels.

Adaptation, Physiological↗

Vegetative growth of higher plants on a three-dimensional clinostat.

Seedlings of rice, maize, cress, pea, and azuki bean were grown on a three-dimensional clinostat and changes in their vegetative growth processes were analyzed. A balanced relationship among the length or the weight of each organ was observed in these species even on the clinostat. Growth of pea second internodes is supported by the transport of sugars from the cotyledons, which was not influenced by the clinostat rotation. Thus, growth correlation and the translocation of sugars normally occurred even under simulated microgravity conditions. In contrast, morphogenesis was clearly changed by the clinostat rotation. The axiality along the gravity vector disappeared and so seedlings formed themselves into a sphere-like shape on the clinostat. The dorsiventrality was indistinct in growth of maize coleoptiles on the surface of the earth, but the clinostat rotation induced a clear dorsinventral bending. These changes in morphogenesis may influence the long-term growth phenomena and modify the life cycle of higher plants under a microgravity environment.

Biological Transport↗

Simulated microgravity (SMG) and bacteria.

This past century has been a scientific revolution in the understanding of the cell as the basic unit of life. However an immense paucity of knowledge exists on microbial growth, survival, function and structure in space. However, there are significant constraints placed on conducting biological research in space such as time, available stowage space, trained personnel, power requirements, weight and the possibility of accidental microbiological contamination. One Earth-based approach is to use a modification of a clinostat known as a HARV (high-aspect-ratio-vessel; Synthecon Inc., Houston, Texas, USA) to conduct this research. In this note we describe the use of the HARV to examine the effects of randomized microgravity (RMG) on bacterial growth and membrane polarization.

Bacteriological Techniques↗

Thyroid organoid formation in simulated microgravity: influence of keratinocyte growth factor.

The generation of artificial human thyroid tissues in suspension (low-shear environment, present in simulated microgravity [MG] and generated by a rotary cell culture system [RCCS]), was enhanced by increasing medium kinematic viscosity with a (3% v/v) suspension of extracellular matrix (basement membrane extract [BME]) in serum-free medium to generate artificial human thyroid organoids. Recombinant human keratinocyte growth factor (KGF, 7 ng/mL) facilitated human thyrocyte aggregation and three-dimensional (3-D) differentiation. There was an MG-associated decrease in extractable DNA that was reversed after addition of keratinocyte growth factor (KGF). In simulated MG, the increase in extractable DNA after KGF addition was up to 170% over non-KGF control cultures. In contrast, monolayer cultures in unit gravity showed a maximum DNA increase of 39% after KGF addition. Morphologically, differentiated thyroid neofollicles displayed polarization and were located in close proximity after 2 weeks of culture. Immunogold labeling with antibody to human thyroglobulin (Tg) revealed staining of follicular lumina and secretory vesicles, and a time-dependent increase in human Tg was detected in the culture media. Culture under simulated MG thus allowed direct visualization of KGF-facilitated thyrocyte/extracellular matrix interaction. Such artificial human thyroid organoids-generated in MG and in the presence of KGF-structurally resembled natural thyroid tissue. The above findings may have implications for autoimmune thyroid disease where KGF (if, for example, secreted locally by intraepithelial gammadelta T cells among other cells) may contribute to thyroid cell growth.

Cell Adhesion Molecules↗

Venoconstrictive thigh cuffs impede fluid shifts during simulated microgravity.

BACKGROUND: This study determined the efficacy of venoconstrictive thigh cuffs, inflated to 50 mmHg, on impeding fluid redistributions during simulated microgravity. METHODS: There were 10 healthy male subjects who were exposed to a 2-h tilt protocol which started in the standing position, and was followed by 30 min supine, 30 min standing, 30 min supine, 30 min of -12 degrees head down tilt (HDT, to simulate microgravity), 15 min of HDT with venoconstrictive thigh cuffs inflated, a further 10 min of HDT, 5 min supine, and 10 min standing. To increase the sensitivity of the techniques in an Earth-based model, 12 degrees HDT was used to simulate microgravity effects on body fluid shifts. Volume changes were measured with anthropometric sleeve plethysmography. RESULTS: Transition to the various tilt positions resulted in concomitant decrements in leg volume (Stand [STD] to Supine [SUP], -3.0%; SUP to HDT, -2.0%). Inflation of the venoconstrictive thigh cuffs to 50 mmHg, during simulated microgravity, resulted in a significant 3.0% increase in leg volume from that seen in HDT (p < 0.01). No significant changes in systemic cardiovascular parameters were noted during cuff inflation. CONCLUSIONS: We conclude that venoconstrictive thigh cuffs, inflated to 50 mmHg for 15 min during 12 degrees HDT, can create a more Earth-like fluid distribution. Cuffs could potentially be used to ameliorate the symptoms of cephalad edema seen with space adaptation syndrome and to potentiate existing fluid volume countermeasure protocols.

Adult↗

Effect of simulated microgravitation on phytohormones and cell structure of tropical orchids.

When studying the effect of two month clinostating on the phytohormonal system of orchids with different types of shoot system branching and different shoot morphology, it was determined that, as a result of simulated microgravitation, endogenous growth regulators changed less in the species with sympodial branching than in species with monopodial branching and without pseudobulbs. Stimulators prevail in the balance of growth regulators in species of the first type and inhibitors in species of the second type. Besides this, comparative analysis of structural organization of juvenile leaf surface tissue of tested orchids was carried out. Variability of size, number and structure of stomatal organization were found according to species belonging to each branching type after clinostating. Electronic microscope studies show some structural peculiarities of epidermal and mesophilous cells.

Gibberellins↗

[Analysis and improvement consideration of the current human experimental models of humoral regulation in microgravity].

The main differences between physiological effects of microgravity in spaceflight and simulated microgravity on humans appeared in the circulation of the low pressure side, in humoral and electrolyte metabolism. For a further understanding of the physiological effects of microgravity, some improvement of the current human experimental models are needed. It is possible to choose more adequate models closer to the situation in microgravity through measuring cardiovascular parameters, fluid regulation and renal excretion variables under head-up tilt (HUT) plus lower body positive pressure condition, or under head-down tilt (HDT) plus upper body negative pressure in various angles and pressure levels.

Adaptation, Physiological↗

The growth movement in the peduncle of Eichhornia crassipes II.

The peduncle of water hyacinth (Eichhornia crassipes) showed the downward bending within 24 hours after full flowering. Previously it was suggested that the downward bending of peduncle might be induced by the differential growth of the epidermal cells of the portion because of the differential distribution of auxin in the upper side of the bending part of the peduncle. In order to investigate the effect of auxin and gravity on the peduncle bending in Water hyacinth, we examined the growth reaction of peduncle and the effects of plant hormones on the bending of peduncle under simulated microgravity, and the sedimentable amyloplast on earth and three dimensional (3D)-clinostat. As a result it was confirmed that the downward bending of peduncle in water hyacinth is the positive gravitropism, and that its phenomenon is caused by the differential distribution of auxin in the upper side of bending part of peduncle. It was found that the amyloplast sediments toward gravity direction in the bending part of the peduncle. From the present results, any direct relation between the sedimentable amyloplast and auxin transport were not cleared in the peduncle of water hyacinth. Further study should be carried out.

Biological Transport↗

Is water immersion useful for analyzing gravity resistance responses in terrestrial plants?

Water immersion has been used as a simulator of microgravity for analyzing gravity responses in semiaquatic plants such as rice. To examine whether or not water immersion for a short experimental period is a useful microgravity simulator even in terrestrial plants, we analyzed effects of water immersion on the cell wall rigidity and metabolisms of its constituents in azuki bean epicotyls. The cell wall rigidity of epicotyls grown underwater was significantly lower than that in the control. Water immersion also caused a decrease in molecular mass of xyloglucans as well as the thinning of the cell wall. Such changes in the mechanical and chemical properties of the cell wall underwater were similar to those observed in microgravity conditions in space. These results suggest that water immersion for a short period is a useful system for analyzing gravity resistance responses even in terrestrial plants.

Cell Wall↗

The exercise and environmental physiology of extravehicular activity.

Extravehicular activity (EVA), i.e., exercise performed under unique environmental conditions, is indispensable for supporting daily living in weightlessness and for further space exploration. From 1965-1996 an average of 20 h x yr(-1) were spent performing EVA. International Space Station (ISS) assembly will require 135 h x yr(-1) of EVA, and 138 h x yr(-1) is planned for post-construction maintenance. The extravehicular mobility unit (EMU), used to protect astronauts during EVA, has a decreased pressure of 4.3 psi that could increase astronauts' risk of decompression sickness (DCS). Exercise in and repeated exposure to this hypobaria may increase the incidence of DCS, although weightlessness may attenuate this risk. Exercise thermoregulation within the EMU is poorly understood; the liquid cooling garment (LCG), worn next to the skin and designed to handle thermal stress, is manually controlled. Astronauts may become dehydrated (by up to 2.6% of body weight) during a 5-h EVA, further exacerbating the thermoregulatory challenge. The EVA is performed mainly with upper body muscles; but astronauts usually exercise at only 26-32% of their upper body maximal oxygen uptake (VO2max). For a given ground-based work task in air (as opposed to water), the submaximal VO2 is greater while VO2max and metabolic efficiency are lower during ground-based arm exercise as compared with leg exercise, and cardiovascular responses to exercise and training are also different for arms and legs. Preflight testing and training, whether conducted in air or water, must account for these differences if ground-based data are extrapolated for flight requirements. Astronauts experience deconditioning during microgravity resulting in a 10-20% loss in arm strength, a 20-30% loss in thigh strength, and decreased lower-body aerobic exercise capacity. Data from ground-based simulations of weightlessness such as bed rest induce a 6-8% decrease in upper-body strength, a 10-16% loss in thigh extensor strength, and a 15-20% decrease in lower-body aerobic exercise capacity. Changes in EVA support systems and training based on a greater understanding of the physiological aspects of exercise in the EVA environment will help to insure the health, safety, and efficiency of working astronauts.

Arm↗

Influence of simulated microgravity environmental factor on ginseng cell growth and ginseng saponin content.

When ginseng (Panax ginseng C.A. Meyer) cells were subjected horizontal rotation on a clinostat, their growth and ginseng saponin content differed from those cultured in normal gravitational environments (control). Both fresh and dry weights of ginseng cells rotating on clinostat were higher than those of the control, and the difference in dry weight was particularly obvious. After 3 weeks of cultivation, saponin content in ginseng was 10% higher under the horizontal rotation treatment on the clinostat than that of the control. When ginseng cells were cultured on Ca2(+)-deprived medium and clinostatted for 3 weeks, their ginseng saponin content was almost twice of that of the control. Besides, in our experiments, the higher the Ca2+ concentration in medium, the lower the ginseng saponin content in the ginseng cells cultured.

Biomass↗

Sampling of prenatal and postnatal offspring from individual rat dams enhances animal use without compromising development.

To assess prenatal and postnatal developmental status in the offspring of a group of animals, it is typical to examine fetuses from some of the dams as well as infants born to the remaining dams. Statistical limitations often arise, particularly when the animals are rare or especially precious, because all offspring of the dam represent only a single statistical observation; littermates are not independent observations (biologically or statistically). We describe a study in which pregnant laboratory rats were laparotomized on day 7 of gestation (GD7) to ascertain the number and distribution of uterine implantation sites and were subjected to a simulated experience on a 10-day space shuttle flight. After the simulated landing on GD18, rats were unilaterally hysterectomized, thus providing a sample of fetuses from 10 independent uteruses, followed by successful vaginal delivery on GD22, yielding postnatal samples from 10 uteruses. A broad profile of maternal and offspring morphologic and physiologic measures indicated that these novel sampling procedures did not compromise maternal well-being and maintained normal offspring development and function. Measures included maternal organ weights and hormone concentrations, offspring body size, growth, organ weights, sexual differentiation, and catecholamine concentrations.

Adrenal Glands↗

Simulated microgravity impairs leukemic cell survival through altering VEGFR-2/VEGF-A signaling pathway.

Motile cells capable of undergoing transendothelial migration, such as hematopoietic and leukemic cells, have been shown to sense and respond to a decrease in their surrounding gravity. In this study, we investigated the effects of microgravity on human leukemic cell proliferation and expression of receptors that control cell survival, such as the tyrosine kinase vascular endothelial growth factor receptor-2 (VEGFR-2). VEGFR-2 is shuttled between the nucleus and membrane, and through an autocrine activation of its ligand, VEGF-A, conveys signals that control cell survival. Autocrine or paracrine stimulation of VEGFR-2 facilitates localization of this receptor from the membrane to the nucleus--a process that results in increased survival of the leukemic cells. Here, we provide evidence that the mechanical forces altered by simulated microgravity localize and maintain VEGFR-2 in the membrane, and also block VEGF-A expression. This interferes with the shuttling of VEGFR-2 to the nucleus, resulting in a decrease in signaling and enhanced leukemic cell death. These data suggest that microgravity modulates cell survival through altering the cellular trafficking and activation state of tyrosine kinase receptors. This study has potential implications for understanding the regulation of receptor biology in pathophysiology, particularly VEGFR trafficking, thereby providing for the development of appropriate therapeutic strategies to abrogate intracrine stimulation triggered by VEGFR internalization.

Cell Proliferation↗