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PubMed2004

Altered cytokine expression in tissues of mice subjected to simulated microgravity.

Space flight is known to induce microgravity-associated immune dysfunction in humans, non-human primates and rodents. To understand the mechanism underlying these defects, several studies in rodents have been conducted in a ground-based antiorthostatic suspension (AOS) model that would mimic the effects of microgravity. In all these in vivo studies that showed the effects on cytokine profiles actually investigated the ex vivo production from culturing the cells isolated from whole organism that was exposed to space flight and/or microgravity. So, the purpose of the study was to examine the in vivo expression of cytokines in mice in immunologically important tissue environments of mice that were subjected to AOS. Cytokines such as Interleukin-1beta, (IL-1beta), IL-2, IL-3, IL-6, Interferon-gamma (IFN-gamma) and Tumor Necrosis Factor-alpha (TNF-alpha) were measured by Enzyme Linked Immunosorbent Assay (ELISA) in the homogenates of spleen tissue, lymph nodes and also in serum of AOS mice and compared with that of control mice. AOS induced no change in the IL-3 levels, but IL-1beta was increased significantly whereas IL-2 levels decreased in spleen, lymph nodes and serum. IL-6 levels did not differ in spleen but were significantly increased in lymph nodes and serum of AOS mice. IFN-gamma levels in spleen did not change but showed nonsignificant reduction in lymph nodes and significant reduction in serum in response to AOS. TNF-alpha levels in spleen and serum were unchanged and increased in lymph nodes. This in vivo cytokine study confirms the earlier findings that microgravity-simulated conditions induce tissue-specific immune response.

Animals↗
PubMed2002

Effects of simulated microgravity on arterial nitric oxide synthase and nitrate and nitrite content.

The aim of the present work was to investigate the alterations in nitric oxide synthase (NOS) expression and nitrate and nitrite (NOx) content of different arteries from simulated microgravity rats. Male Wistar rats were randomly assigned to either a control group or simulated microgravity group. For simulating microgravity, animals were subjected to hindlimb unweighting (HU) for 20 days. Different arterial tissues were removed for determination of NOS expression and NOx. Western blotting was used to measure endothelial NOS (eNOS) and inducible NOS (iNOS) protein content. Total concentrations of NOx, stable metabolites of nitric oxide, were determined by the chemiluminescence method. Compared with controls, isolated vessels from simulated microgravity rats showed a significant increase in both eNOS and iNOS expression in carotid arteries and thoracic aorta and a significant decrease in eNOS and iNOS expression of mesenteric arteries. The eNOS and iNOS content of cerebral arteries, as well as that of femoral arteries, showed no differences between the two groups. Concerning NOx, vessels from HU rats showed an increase in cerebral arteries, a decrease in mesenteric arteries, and no change in carotid artery, femoral artery and thoracic aorta. These data indicated that there were differential alterations in NOS expression and NOx of different arteries after hindlimb unweighting. We suggest that these changes might represent both localized adaptations to differential body fluid redistribution and other factors independent of hemodynamic shifts during simulated microgravity.

Animals↗
PubMed2005

Exercise within lower body negative pressure partially counteracts lumbar spine deconditioning associated with 28-day bed rest.

Astronauts experience spine deconditioning during exposure to microgravity due to the lack of axial loads on the spine. Treadmill exercise in a lower body negative pressure (LBNP) chamber provides axial loads on the lumbar spine. We hypothesize that daily supine LBNP exercise helps counteract lumbar spine deconditioning during 28 days of microgravity simulated by bed rest. Twelve sets of healthy, identical twins underwent 6 degrees head-down-tilt bed rest for 28 days. One subject from each set of twins was randomly assigned to the exercise (Ex) group, whereas their sibling served as a nonexercise control (Con). The Ex group exercised in supine posture within a LBNP chamber for 45 min/day, 6 days/wk. All subjects underwent magnetic resonance imaging of their lumbar spine before and at the end of bed rest. Lumbar spinal length increased 3.7 +/- 0.5 mm in the Con group over 28-day bed rest, whereas, in the Ex group, lumbar spinal length increased significantly less (2.3 +/- 0.4 mm, P = 0.01). All lumbar intervertebral disk heights (L5-S1, L4-5, L3-4, L2-3, and L1-2) in the Con group increased significantly over the 28-day bed rest (P < 0.05). In the Ex group, there were no significant increases in L5-S1 and L4-5 disk heights. Lumbar lordosis decreased significantly by 3.3 +/- 1.2 degrees during bed rest in the Con group (P = 0.02), but it did not decrease significantly in the Ex group. Our results suggest that supine LBNP treadmill exercise partially counteracts lumbar spine lengthening and deconditioning associated with simulated microgravity.

Adaptation, Physiological↗
PubMed1995

Effects of clinorotation and microgravity on sweet clover columella cells treated with cytochalasin D.

The cytoskeleton of columella cells is believed to be involved in maintaining the developmental polarity of cells observed as a reproducible positioning of cellular organelles. It is also implicated in the transduction of gravitropic signals. Roots of sweet clover (Melilotus alba L.) seedlings were treated with a microfilament disrupter, cytochalasin D, on a slowly rotating horizontal clinostat (2 rpm). Electron micrographs of treated columella cells revealed several ultrastructural effects including repositioning of the nucleus and the amyloplasts and the formation of endoplasmic reticulum (ER) whorls. However, experiments performed during fast clinorotation (55 rpm) showed an accumulation (but no whorling) of a disorganized ER network at the proximal and distal pole and a random distribution of the amyloplasts. Therefore, formation of whorls depends upon the speed of clinorotation, and the overall impact of cytochalasin D suggests the necessity of microfilaments in organelle positioning. Interestingly, a similar drug treatment performed in microgravity aboard the US Space Shuttle Endeavour (STS-54, January 1993) caused a displacement of ER membranes and amyloplasts away from the distal plasma membrane. In the present study, we discuss the role of microfilaments in maintaining columella cell polarity and the utility of clinostats to simulate microgravity.

Actin Cytoskeleton↗
PubMed2001

Simulated microgravity induces alteration in the central nervous system.

To investigate whether the signs of neurophysiological impairment observed in flight may be traced back to cytomorphology, we undertook a ground-based study focusing upon the architecture of cultured glial cells under simulated microgravity obtained by three-dimensional clinorotation.

Actin Cytoskeleton↗
PubMed2004

Effects of clinorotation on COL1A1- EGFP gene expression.

Bone-formation related gene plays a critical role in bone loss induced by space microgravity, however the exact mechanism is unclear. In this study, we aim to investigate the effect of microgravity on the activity of alpha 1(I) collagen (COL1A1) gene promoter and the expression of osteoblast-related genes. COL1A1 promoter was digested by restriction enzymes resulting in three DNA fragments. The fragments were ligated with the enhanced green fluorescent protein report gene, and subcloned into expression vectors. ROS17/2.8 cells transfected by these vectors were screened by G418, and enhanced green fluorescent protein (EGFP) positive colonies were isolated and cultured under clinostat condition. EGFP and Collagen type I expression level were detected by fluorescence intensity analysis and immunocytochemistry methods respectively. The results showed that the expression of EGFP and collagen type I was increased 24 h, 48 h after the cells were cultured under stimulated microgravity, illustrating that the activity of COL1A1 promoter might be increased. In conclusion, osteoblasts can compensatively increase the expression of type I collagen by enhancing the activity of COL1A1 promoter under short-term simulated microgravity conditions.

Animals↗
PubMedDate not supplied

Ion regulatory function of the human kidney in prolonged space flights.

Ten cosmonauts, who performed 30-175-day space flights aboard Salyut-4 and Salyut-6, and over 60 test subjects who were exposed to bed rest of up to 182 days and immersion of up to 56 days, were examined. The renal excretion of potassium and calcium increased, reaching a maximum by the 4-6th weeks in prolonged space flights and simulation studies. During the load tests with potassium and calcium salt, excretion postflight was much higher than preflight. During potassium chloride load tests a positive correlation between the blood content of aldosterone and potassium excretion existed, whereas during calcium lactate load tests an increased calcium excretion was accompanied by a decrease in blood parathyroid hormone concentration. The most probable cause of the negative ion balance in weightlessness is the reduced capacity of tissues to retain electrolytes due to the decreased ion pool capacity. Different exercises have been shown to exert a beneficial effect on electrolyte metabolism.

Aldosterone↗
PubMed1998

Simulated microgravity inhibits the genetic expression of interleukin-2 and its receptor in mitogen-activated T lymphocytes.

Experiments conducted in space in the last two decades have shown that T lymphocyte activation in vitro is remarkably reduced in microgravity. The data indicate that a failure of the expression of the interleukin-2 receptor (measured as protein secreted in the supernatant) is responsible of the loss of activity. To test such hypothesis we have studied the genetic expression of interleukin-2 and of its receptor in concanavalin A-activated lymphocytes with the RT-PCR technology. Microgravity conditions were simulated in the fast rotating clinostat and in the random positioning machine. The latter is an instrument introduced recently to study gravitational effects on single cells. Our data clearly show that the expression of both IL-2 and IL-2Ralpha genes is significantly inhibited in simulated O X g. Thus full activation is prevented.

Biophysics↗
PubMed2000

Power spectral analysis imperfectly informs changes in sympathetic traffic during acute simulated microgravity.

The purpose of this study was to investigate the value of frequency-domain analysis of autonomic rhythms as a simple, non-invasive technique for the study of immediate neural adjustments to simulated microgravity. We continuously recorded the electrocardiogram, non-invasive beat-by-beat arterial pressure, and muscle sympathetic nerve activity (MSNA) during 5-min periods of controlled frequency breathing (15 breaths.min-1) with subjects (n = 10) in supine, and 10 degrees head-down tilt positions. We estimated changes in fluid volume with lower leg circumference measurements. We analyzed data in the frequency domain with fast Fourier-based power spectral analysis, and calculated the ratio of normalized low-to-respiratory frequency RR-interval spectral power as an index of sympathetic activity. Head-down tilt significantly reduced lower leg volume, MSNA, and MSNA oscillations at the respiratory frequency (p < 0.05). Head-down tilt did not change RR-interval, arterial pressure, or their power spectra (p > 0.05). We conclude that non-invasive frequency-domain estimates do not adequately reveal subtle changes in sympathetic traffic during acute, simulated microgravity.

Adaptation, Physiological↗
PubMed2001

[Effect of lower body negative pressure and rotating-table simulated push-pull effect in flight on cardiovascular function].

Objective. To explore the effect of push-pull effect (PPE) simulated by lower body negative pressure (LBNP) rotating-table, and observe the physiological responses to push-pull maneuver. Method. A special LBNP rotating-table was used to simulate the push-pull maneuver. 8 healthy adults participated randomly in two experiments. One was simulated PPE test, which include a series of head-up stand (HUT, +1 Gz) for 1 min, then in head-down stand (HDT, -1 Gz) for 30 s and again in HUT combining LBNP (-50 mmHg) for 10 min. The other one was control test, which only consists of HUT combining LBNP (-50 mmHg) for 10 min. Changes of heart rate (HR), blood pressure (BP), basic impedance (Z0) , stroke output (SO) , cardiac output (CO) and total peripheral resistance (TPR) were monitored by electrical impedance instrument during the experiment. Result. During simulated PPE experiment, 3 subjects presented presyncopal symptoms, with average standing time of 8.99 +/- 1.47 min, while during control test, all the subjects completed HUT combining LBNP for 10 min. In simulated PPE experiment, as compared with HUT (control), HR, Z0 during HDT were significantly lowered, while SV and CO were increased significantly. During HUT + LBNP, HR, Z0 and TPR were significantly higher, while SV and CO were significantly lower than that of control and HDT. SBP was increased significantly than control value when "HUT + LBNP" started, but during the whole process of "HUT + LBNP ", it became significantly lowered. In control experiment, the above mentioned indexes showed the same trend of as change compared with the control, however, the percentage of the change was lower than simulated PPE test, the change percentage in HR was not including. Conclusion. After headstand, head-up stand combining LBNP caused cardiovascular function descends, the degree was larger than simple head-up stand combining LBNP. LBNP rotating-table can be used to simulated push-pull effect.

Adult↗
PubMed2003

Gravitational and hormonal control in secondary xylem formation of Japanese flowering cherry.

It has been reported that Japanese flowering cherry promotes the formation of tension wood in the upper side of the inclined stem, which induces the negative gravitropism. In the other hand, the plant under simulated micro-gravity conditions reduces the width of the secondary xylem and increases the density of the vessels, which means decreased the density of the fiber cells. The plant under simulated micro-gravity conditions showed that the inhibition of xylem development and the decreased supporting mechanism in the stem. In this study we examined that the effects of auxin and gibberellin in the plant under simulated micro-gravity conditions and inclined stimulus. The gibberellin treatment promotes the secondary xylem development under simulated microgravity conditions and inclined stimulus. The upper side of the inclined stem contents much higher levels of IAA and gibberellin A1(GA1) than the lower side of that.

Gibberellins↗
PubMed1980

Femur-bending properties as influenced by gravity: IV. Limits after high and low weight-bearing.

Gravity enhances femur growth as measured in terms of strength sigma u, but shows little or even a growth-retarding effecting in terms of "relative brittleness," defined as the inverse 1/epsilon u of ultimate or tolerable strain. Chronic weightlessness was simulated by harness suspension or by extrapolation of results from 3-G centrifugation. Experimental results from 45 male, white rats (34-520 d old) were compared to 72 control or baseline rats (28-520 d old) white correction for age and size differences. After suspension, the youngest rats showed subnormal epsilon u. Combined results, however, although predicting 19 +/- 1% below normal sigma u, after a week of weightlessness, predicted less effect (1 +/- 4%) for epsilon u.

Animals↗
PubMed2004

Sleep restriction does not affect orthostatic tolerance in the simulated microgravity environment.

Orthostatic intolerance (OI) is a major problem following spaceflight, and, during flight, astronauts also experience sleep restriction. We hypothesized that sleep restriction will compound the risk and severity of OI following simulated microgravity and exaggerate the renal, cardioendocrine, and cardiovascular adaptive responses to it. Nineteen healthy men were equilibrated on a constant diet, after which they underwent a tilt-stand test. They then completed 14-16 days of simulated microgravity [head-down tilt bed rest (HDTB)], followed by repeat tilt-stand test. During HDTB, 11 subjects were assigned to an 8-h sleep protocol (non-sleep restricted), and 8 were assigned to a sleep-restricted protocol with 6 h of sleep per night. During various phases, the following were performed: 24-h urine collections, hormonal measurements, and cardiovascular system identification. Development of presyncope or syncope defined OI. There was a significant decrease in time free of OI (P = 0.02) and an increase in OI occurrence (P = 0.06) after HDTB among all subjects. However, the increase in OI occurrence did not differ significantly between the two groups (P = 0.60). The two groups also experienced similar physiological changes with HDTB (initial increase in sodium excretion; increased excretion of potassium at the end of HDTB; increase in plasma renin activity secretion without a change in serum or urine aldosterone). No significant change in autonomic function or catecholamines was noted. Simulated microgravity leads to increased OI, and sleep restriction does not additively worsen OI in simulated microgravity. Furthermore, conditions of sleep restriction and nonsleep restriction are similar with respect to renal, cardioendocrine, and cardiovascular responses to simulated microgravity.

Adult↗
PubMed2002

Growth and photosynthesis of Japanese flowering cherry under simulated microgravity conditions.

The photosynthetic rate, the leaf characteristics related to photosynthesis, such as the chlorophyll content, chlorophyll a/b ratio and density of the stomata, the leaf area and the dry weight in seedlings of Japanese flowering cherry grown under normal gravity and simulated microgravity conditions were examined. No significant differences were found in the photosynthetic rates between the two conditions. Moreover, leaf characteristics such as the chlorophyll content, chlorophyll a/b ratio and density of the stomata in the seedlings grown under the simulated microgravity condition were not affected. However, the photosynthetic product of the whole seedling under the simulated microgravity condition increased compared with the control due to its leaf area increase. The results suggest that dynamic gravitational stimulus controls the partitioning of the products of photosynthesis.

Chlorophyll↗
PubMed2004

Daily short-period gravitation can prevent functional and structural changes in arteries of simulated microgravity rats.

This study was designed to clarify whether simulated microgravity-induced differential adaptational changes in cerebral and hindlimb arteries could be prevented by daily short-period restoration of the normal distribution of transmural pressure across arterial vasculature by either dorsoventral or footward gravitational loading. Tail suspension (Sus) for 28 days was used to simulate cardiovascular deconditioning due to microgravity. Daily standing (STD) for 1, 2, or 4 h, or +45 degrees head-up tilt (HUT) for 2 or 4 h was used to provide short-period dorsoventral or footward gravitational loading as countermeasure. Functional studies showed that Sus alone induced an enhancement and depression in vasoconstrictor responsiveness of basilar and femoral arterial rings, respectively, as previously reported. These differential functional alterations can be prevented by either of the two kinds of daily gravitational loading treatments. Surprisingly, daily STD for as short as 1 h was sufficient to prevent the differential functional changes that might occur due to Sus alone. In morphological studies, the effectiveness of daily 4-h HUT or 1-h STD in preventing the differential remodeling changes in the structure of basilar and anterior tibial arteries induced by Sus alone was examined by histomorphometry. The results showed that both the hypertrophic and atrophic changes that might occur, respectively, in cerebral and hindlimb arteries due to Sus alone were prevented not only by daily HUT for 4 h but also by daily STD even for 1 h. These data indicate that daily gravitational loading by STD for as short as 1 h is sufficient to prevent differential adaptational changes in function and structure of vessels in different anatomic regions induced by a medium-term simulated microgravity.

Adaptation, Physiological↗
PubMed1997

[Observation of changes of cardiovascular function during 2.5h HDT (-15 degrees) with sphygmogram method].

To understand the changes of cardiovascular functions in the initial stage of space flight, the changes in 19 healthy young men during 2.5h head down tilt (HDT) (15 degrees) were observed with CF-II cardiovascular function detecting and diagnosing equipment. The blood pressure and sphygmogram of left radial arterial were recorded in sedentary condition and at 10th, 30th, 60th, 90th, 120th and 140th minute of HDT. The results showed that changes of cardiovascular indices during HDT can be divided into acute regulation stage (< 1h) and the regulation stage (1-2.5 h); circulatory blood volume, stroke volume and cardiac output were increased, while heart rate, pre and after load of the heart, CVP, coronary circulatory function, blood pressure and systemic vascular resistance were decreased; the vagus feedback index increased and the regulation function of vasscule decreased. Most of the changes of cardiovascular indices as reflected in the sphygmogram are in consistent with the reported result in space flight or simulated micro-G, so the sphygmogram method might be applied to space medical research.

Aerospace Medicine↗
PubMed2004

[Selection of scaffolds of rat hepatocytes in three-dimension culture under simulated microgravity].

In order to investigate the biocompatibility, degradation and inflammatory reactions of poly benzyl glutamate (PBLG), poly lactic-co-glycolic acid (PLGA) and fibrin gel used as the scaffolds co-cultured with the hepatocytes of rats under simulated microgravity conditions in rotating wall vessel bioreactor (RWVB). PLGA is low biocompatibility, PBLG is medium biocompatibility with rat hepatocytes. Fibrin gel is high biocompatibility, no toxic degradation and no inflammatory reactions. Fibrin gel can be used as three dimensional scaffolds of rat hepatocytes under simulated microgravity.

Animals↗