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PubMed1997

Skeletal muscle satellite cells cultured in simulated microgravity.

Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and, therefore, provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75% satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D culture and 3-D HARV culture. Plating efficiency (cells attached divided by cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and nonsatellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability because glucose levels in medium from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARV were joined together by cells into 3-D aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of satellite cells on microcarrier beads within the HARV bioreactor results in a 3-D level of organization that could provide a more suitable model to study postnatal muscle development than is currently available with standard culture methods.

Animals↗
PubMed1997

Changes in gravity inhibit lymphocyte locomotion through type I collagen.

Immunity relies on the circulation of lymphocytes through many different tissues including blood vessels, lymphatic channels, and lymphoid organs. The ability of lymphocytes to traverse the interstitium in both nonlymphoid and lymphoid tissues can be determined in vitro by assaying their capacity to locomote through Type I collagen. In an attempt to characterize potential causes of microgravity-induced immunosuppression, we investigated the effects of simulated microgravity on human lymphocyte function in vitro using a specialized rotating-wall vessel culture system developed at the Johnson Space Center. This very low shear culture system randomizes gravitational vectors and provides an in vitro approximation of microgravity. In the randomized gravity of the rotating-wall vessel culture system, peripheral blood lymphocytes did not locomote through Type I collagen, whereas static cultures supported normal movement. Although cells remained viable during the entire culture period, peripheral blood lymphocytes transferred to unit gravity (static culture) after 6 h in the rotating-wall vessel culture system were slow to recover and locomote into collagen matrix. After 72 h in the rotating-wall vessel culture system and an additional 72 h in static culture, peripheral blood lymphocytes did not recover their ability to locomote. Loss of locomotory activity in rotating-wall vessel cultures appears to be related to changes in the activation state of the lymphocytes and the expression of adhesion molecules. Culture in the rotating-wall vessel system blunted the ability of peripheral blood lymphocytes to respond to polyclonal activation with phytohemagglutinin. Locomotory response remained intact when peripheral blood lymphocytes were activated by anti-CD3 antibody and interleukin-2 prior to introduction into the rotating-wall vessel culture system. Thus, in addition to the systemic stress factors that may affect immunity, isolated lymphocytes respond to gravitational changes by ceasing locomotion through model interstitium. These in vitro investigations suggest that microgravity induces non-stress-related changes in cell function that may be critical to immunity. Preliminary analysis of locomotion in true microgravity revealed a substantial inhibition of cellular movement in Type I collagen. Thus, the rotating-wall vessel culture system provides a model for analyzing the microgravity-induced inhibition of lymphocyte locomotion and the investigation of the mechanisms related to lymphocyte movement.

Animals↗
PubMedDate not supplied

Recent advances in the physiology of whole body immersion.

Recent investigations have furnished a complete analysis of the hemodynamic events accompanying whole-body immersion. About 700 ml of blood are translocated into the intrathoracic circulation, and heart volume increases by 180 +/- 62 ml. These changes are followed by an increase in stroke volume and cardiac output of over 30%. At the same time a reflex reduction of total peripheral resistance and venous tone occurs. Renin and aldosterone activity are reduced while the 17-hydroxycorticosteroid is not affected. Treatment of the subject with DOCA attenuates but does not extinguish the excess sodium excretion of immersion. This finding strengthens the arguments in favor of an unknown factor enhancing sodium excretion. Finally, the relative activation of the three factors that serve volume control, the excretory function of the kidney, capillary filtration pressure, and the thirst mechanism, is discussed.

17-Hydroxycorticosteroids↗
PubMedDate not supplied

Bone mineral measurement: Skylab experiment M-078.

The observation that bone mineral is lost in patients who are either immobilized or remain in bed for extended periods of time formed the basis for the concern that large amounts of bone mineral may be lost during long periods of weightlessness. This concern was magnified when early X-ray densitometry studies suggested that rather large amounts of mineral could be lost during rather short periods of weightlessness (4-14 days). Even though these Gemini results have recently been modified, they still reflect substantial losses in the upper extremity. This led to a series of prolonged bed-rest studies (30-36 weeks) which, in addition to careful calcium balance, also employed a newer, more precise method of estimating bone mineral in the radius, ulna, and os calcis. It employed an essentially monoenergetic photon source (125I) and a scintillation detector operating in a rectilinear scanning mode to measure bone mineral by the absorptiometric technique. Bed-rest studies revealed variable mineral losses but suggested that little if any is lost during 4-6 weeks, with variable amounts being lost in 8 weeks. Losses up to 40% were noted in the os calcis after 9 months, with essentially none in the radius and ulna. When this technique was employed during the Apollo 14, 15, and 16 missions, only one crewman (CMP Apollo 15) showed significant losses in the os calcis and none in the radius or ulna. These results were, therefore, in concert with the bed-rest data but at variance with the earlier Gemini data. The variability observed during bed rest was reconciled when it was observed that the rate of loss could be correlated with the initial 24-hour urinary hydroxyproline excretion and the initial os calcis mineral content. Prediction terms were established. Measurements of the SL-II crew after 28 days of weightlessness revealed no significant bone mineral losses. The Skylab data lie within the predicted limits obtained from the bed-rest data. The relevance of the prediction terms to the Skylab and longer missions discussed.

Absorptiometry, Photon↗
PubMedDate not supplied

Characteristics of metabolism during prolonged water immersion.

The effect on the organism of a 12-day stay in a water immersion medium (head on the water surface) was studied on 10 subjects. The condition of the metabolic processes in the subjects was judged from the investigation of the parameters of protein and carbohydrate metabolism, acid-base equilibrium, the activity of a number of enzymes and steroid hormones. The venous blood and diurnal urine served as the material for conducting the corresponding biochemical determinations. The stay in an immersion medium is accompanied by a significant increase in the content of residual nitrogen in the blood, by a reduction of the creatinine content in the blood and by an increase of the creatine content of the blood with the simultaneous increase of the excretion of creatinine and creatine with the urine, by a significant increase of the glucose and lactate content in the blood, by the development of acidosis of a respiratory and metabolic character, as well as by a significant increase of the activity of creatine phosphokinase and the isoenzyme fraction of lactate dehydrogenase3. In all the subjects, an expressed intensification of the glucocorticoid and androgenic functions of the adrenal cortex is noted during the experimental period.

Adaptation, Physiological↗
PubMed1977

Cardiorespiratory responses to exercise after bed rest in men and women.

The purpose of this study was to compare cardiorespiratory responses of men and women to submaximal and maximal workloads before and after bed rest (BR). Fifteen male college students (19-23 yr) and 8 female nurses (23-34 yr) underwent 14 d and 17 d, respectively of bed rest. The maximal work capacity test was performed in the supine position on a bicycle ergometer just before and immediately after bed rest. The women's maximal O2 uptake (maximal VO2) was 41% lower (P<0.05) than the men's before bed rest and 42% lower (P<0.05) after bed rest. During bed rest the women's maximal VO2 decreased from 2.06 to 1.86 liter/min (-9.7%, P<0.05), and that of the men decreased from 3.52 to 3.20 liter/min (-9.1%, P<0.05). Compared with pre-BR values, after bed rest the maximal ventilatory volume was essentially unchanged in the men (+1.8%) and women (+ 2.3%), but maximal heart rate was elevated from 185 to 193 b/min (+ 4.3%, P<0.05) in the men and from 181 to 187 b/min (3.3%, P<0.05) in the women. Submaximal VO2 was unchanged after bed rest in the men but was significantly reduced in the women; the women's Hct and RBC levels were lower (P<0.05) than comparable male data. Mean corpuscular volume was unchanged in both groups pre- and post-bed rest. It is concluded that the proportional deterioration in maximal VO2 following prolonged bed rest was essentially the same in young men and women.

Adult↗
PubMed1979

Simulation of space cabin atmosphere.

The Soyuz 22 space cabin atmosphere was studied for volatile organic trace contaminants. By gas chromatography the following constituents were identified: methane, ethane, heptane, methanol, ethanol, n-propanol, acetaldehyde, acetone, ethyl benzene. Except for acetone, concentrations of the above compounds were close to the values determined in the mock-up experiments.

Air Conditioning↗
PubMed1979

Fluid volumes changes induced by spaceflight.

The blood volume (BV), plasma volume (PV), and extracellular fluid volume changes produced in crewmembers during spaceflights of 11-84 days were compared to changes after 14 or 28 days of bedrest. Spaceflight and bedrest produce approximately equal BV changes but the recorded PV change after spaceflight was less. However, the diurnal change in PV may explain the smaller decreases recorded after spaceflight. The cardiovascular deconditioning caused by spaceflight and bedrest was compared using the mean heart rate response to lower body negative pressure (LBNP) testing at -50 mmHg pressure. These tests show approximately equal LBNP produced heart rate changes after bedrest and spaceflight. A countermeasure which includes 4 hr of LBNP treatment at -30 mmHg and the ingestion of one l. of saline was studied and found capable of returning the heart rate response and the PV of bedrested subjects to control (prebedrest) levels suggesting that it would be useful to the crewmembers after a spaceflight.

Bed Rest↗
PubMed1982

Simple technique to evaluate on the ground the energetic expenditure of physical exercise carried out in weightlessness.

A simple method is described to evaluate the energetic expenditure of graded muscular exercises carried out on the ground in such a way as to approximate the state of space microgravity. Consumption of O2 measured at different rates of execution is shown as well as a tentative computation of the mechanical work on the basis of the accelerations impressed alternately on the mass of the trunk and upper limbs. The practicability of the method proposed under space conditions is pointed out; it does not require any ergometer, in fact, but only the fixing of the subject's feet to the floor of the vehicle.

Adult↗
PubMedDate not supplied

The effect of hypokinesia on lipid metabolism in adipose tissue.

The increase of nonesterified fatty acid (NEFA) concentration in plasma was observed in rats subjected to hypokinesia for 1-60 days. In the period of recovery (7 and 21 days after 60 days immobilization) the content of NEFA returned to control values. The increase of fatty acid release from adipose tissue was observed in hypokinetic rats, however the stimulation of lipolysis by norepinephrine was lower in rats exposed to hypokinesis. The decrease of the binding capacity and a diminished number of beta-adrenergic receptors were found in animals after hypokinesia. The augmentation of the incorporation of glucose into lipids and the marked increase in the stimulation of lipogenesis by insulin were found in adipose tissue of rats subjected to long-term hypokinesia. These results showed an important effect of hypokinesia on lipid mobilization, on lipogenesis and on the processes of hormone regulation in adipose tissue.

Adipocytes↗
PubMed1990

Selective hypergravity stimulation: its effects on the human balance and gait functions. A model to assess, in normal gravity conditions, some aspects of the perturbations induced on human body by microgravity conditions.

To assess on Earth some reactions of the muscle mechanoreceptors to transitions from normogravity to microgravity, we studied the effects of transitions from hypergravity to normogravity. Hypergravity was selectively applied to the extensor muscles by increasing their activity during half an hour by means of 2 rubber extensible springs stretched from shoulders to feet. Immediate effects and post effects of such a stimulation were measured on quantifying dynamic balance (angular or linear displacement) and gait functions (spatio-temporal parameters and inferior limb length variations). The main results are: (1) a post effect on the balance function, appearing 3 minutes after the end of the selective hypergravity stimulus and improving the efficiency of balance function compared with the basal one, (2) a post effect on the gait function, appearing immediately after the end of the selective stimulation. It concerns the measures which quantify the gait phases during which flexor muscles are active (swing phases). It decreases the efficiency of the gait function compared with the basal one, It disappears 3 minutes after the end of the selective hypergravity stimulation. According to these results, if the effects on the muscle mechanoreceptors of the transitions from normogravity, to microgravity looks like those of transitions from hypergravity to normogravity, post effects could be a mechanism of the motor perturbations at the beginning of the orbital flights.

Adult↗
PubMed1991

Effects of saline loading during head down tilt on ANP and cyclic GMP levels and on urinary fluid excretion.

In the present study the renal and humoral effects of acute saline infusions were investigated in six healthy male volunteers before, during and after a ten day period of -6 degrees head-down-tilt (HDT). During the whole 23-day study period the subjects received a standardized diet including 40 ml water and 125 mg NaCl per kg body weight per day. After the infusion of 0.9% saline (22 ml/kg within 20 minutes) plasma atrial natriuretic peptide (ANP) levels were only slightly increased (not significant) at the end of the infusion, while plasma cyclic GMP levels were significantly increased by about 40% (p<0.05) one hour later. No difference was observed in the plasma ANP and cyclic GMP changes between the pre-HDT, the HDT and the post-HDT infusion experiment. Urine flow, sodium excretion and urinary cyclic GMP excretion were significantly increased (p<0.05 and below) by 100 to 300% during the second and third hour after each saline infusion. However, during these short-term periods only 20% of the infused water and less than 20% of the infused sodium were excreted. Furthermore, a significantly increased volume, sodium and cyclic GMP excretion was observed for over 48 hours after each fluid load experiment. These data suggest that HDT does not induce major alterations in the regulation of an acute saline infusion and plasma ANP does not play a major role in the diuretic/natriuretic effects of volume loading.

Adult↗
PubMed1991

The involvement of cerebrovascular reactivity in pathogenesis of space motion sickness.

The aim of the presented work is to investigate the mechanisms of involvement of cerebral circulation in pathogenesis of space motion sickness. In experiments on chronic rabbits the 2-hours rocking and antiorthostasis served as a model of influences of the zero-gravity conditions intracranial hemo- and liquorodynamics. Cerebravascular reactivity was evaluated as a response of cerebral blood flow and intracerebral rheoencephalography to functional loads (photostimulation, CO2-inhalation, Stookey test). Shifts of water balance in the brain tissue has been assessed by means of three-frequency brain electrical impedance. Rocking plus aniorthostatis resulted in decrease of the cerebrovascular reactivity and hyperhydration. The data obtained show a significant decrease of compensatory abilities of the cerebrovascular system under the zero-gravity conditions.

Animals↗
PubMed1992

Electrical vestibular stimulation and space motion sickness.

Electrical vestibular stimulation (EVS) in dynamic balance condition was studied in order to search for a new provocative test of space motion sickness (SMS). SMS is usually attributed to a sensory conflict caused by exposure to microgravity. Vestibular information is conflicting but also unusual and insignificant. EVS is in accordance with this feature because it is not the adequate stimulus of the vestibular receptors. EVS was achieved by means of binaural electrical stimulation. Effects of EVS were potentiated by compelling the subject to maintain dynamic balance on a seesaw. The quantification of this function was performed before during and after EVS in order to investigate a possible relationship between objective consequences of EVS i.e. dynamic balance disturbances, and the discomfort experienced by the subjects. Dynamic balancing skill was statistically worsened during EVS. Moreover EVS evoked subjective symptoms of SMS in 17 out of the 30 subjects examined. During EVS in eyes open conditions, the subjects who encountered the strongest discomfort, presented the most disturbed dynamic balance, evidencing a relationship between the level of discomfort and the imbalance arising from EVS This method could thus constitute an interesting basis of SMS ground-based test.

Adult↗
PubMed1994

Impact of skeletal unloading on bone formation: role of systemic and local factors.

We have developed a model of skeletal unloading using growing rats whose hindlimbs are unweighted by tail suspension. The bones in the hindlimbs undergo a transient cessation of bone growth; when reloaded bone formation is accelerated until bone mass is restored. These changes do not occur in the normally loaded bones of the forelimbs. Associated with the fall in bone formation is a fall in 1,25(OH)2D3 production and osteocalcin levels. In contrast, no changes in parathyroid hormone, calcium, or corticosterone levels are seen. To examine the role of locally produced growth factors, we have measured the mRNA and protein levels of insulin like growth factor-1 (IGF-1) in bone during tail suspension. Surprisingly, both the mRNA and protein levels of IGF-1 increase during tail suspension as bone formation is reduced. Furthermore, the bones in the hindlimbs of the suspended animals develop a resistance to the growth promoting effects of both growth hormone and IGF-1 when given parenterally. Thus, the cessation of bone growth with skeletal unloading is apparently associated with a resistance to rather than failure to produce local growth factors. The cause of this resistance remains under active investigation.

Animals↗