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A new application of LBNP to measure orthostatic tolerance before and after 0-g simulation (water immersion).

Lower body negative pressure (LBNP) remains an important device for the generation of orthostatic stress in the space flight environment as well as a tool to measure inflight and postflight changes in orthostatic response. These applied levels of LBNP have typically not exceeded 50-60 mm Hg negative pressure. Information is incomplete as to the levels of absolute LBNP orthostatic tolerance, and the factors responsible for their variance. A better definition of the tolerance limits for males and females could be expected to aid the evaluation of lower levels of LBNP. An LBNP device was built to study absolute orthostatic tolerance; additionally, another LBNP device was constructed to permit orthostatic tolerance testing directly after a controlled water immersion period. Absolute LBNP orthostatic tolerance patterns are analyzed for a group of males and females (series I). A preliminary statement on the variations of LBNP orthostatic tolerance after limited periods of water immersion and bed rest is also provided (series II).

Adolescent↗

Bone biochemistry in rat femoral diaphysis after space flight.

The aim of this experiment was to identify the location of the biochemical changes associated with depressed mineralization during space flight. We carried out biochemical analysis of 4 sections of the femoral diaphyses from 107 day old male rats flown aboard Cosmos 2044 Biosatellite for 16 days. Control femurs were preflight, vivarium, synchronous for feed, cage and temperature exposure, and a flight simulation model. Distal sections in both the flight and synchronous femurs showed mineral deficits associated with reduced levels of the reducible cross-link product of type I collagen, dehydro-dihydroxylysinonorleucine (deH-DHLNL) (p<.05). Unloaded bones in the ground based flight simulation model showed changes in cross-links similar to flight and synchronous controls, but were not associated with the mineral deficit. Mean values of elements measured in each section of all groups revealed significant associations (p<.005) between the non-collagenous protein, osteocalcin and calcium (r=0.774), phosphorus (r=-.624) and deH-DHLNL/deH-HLNL (r=.883). The ratio of the nonreducible cross-link, pyridinoline, to its lysl analogue, deoxypyridinoline, was consistently lower in the distal than proximal sections of the groups tested. None of the changes during space flight were unique to flight bone. The most significant and extensive changes in bone composition, i.e. mineral deficits associated with changes in both osteocalcin and reducible cross-links, were located in the distal section of the diaphysis of the femur.

Amino Acids↗

Enhancement of phototropic response to a range of light doses in Triticum aestivum coleoptiles in clinostat-simulated microgravity.

The phototropic dose-response relationship has been determined for Triticum aestivum cv. Broom coleoptiles growing on a purpose-built clinostat apparatus providing gravity compensation by rotation about a horizontal axis at 2 rev min-1. These data are compared with data sets obtained with the clinostat axis vertical and stationary, as a 1 g control, and rotating vertically to examine clinostat effects other than gravity compensation. Triticum at 1 g follows the well-established pattern of other cereal coleoptiles with a first positive curvature at low doses, followed by an indifferent response region, and a second positive response at progressively increasing doses. However, these response regions lie at higher dose levels than reported for Avena. There is no significant difference between the responses observed with the clinostat axis vertical in the rotating and stationary modes, but gravity compensation by horizontal rotation increases the magnitude of first and second positive curvatures some threefold at 100 min after stimulation. The indifferent response is replaced by a significant curvature towards the light source, but remains apparent as a reduced curvature response at these dose levels.

Cotyledon↗

Proliferation of human hematopoietic bone marrow cells in simulated microgravity.

Expansion and/or maintenance of hematopoietic stem cell (HSC) potential following in vitro culture remains a major obstacle in stem cell biology and bone marrow (BM) transplantation. Several studies suggest that culture of mammalian cells in microgravity (micro-g) may reduce proliferation and differentiation of these cells. We investigated the application of these findings to the field of stem cell biology in the hopes of expanding HSC with minimal loss of hematopoietic function. To this end, BM CD34+ cells were cultured for 4-6 d in rotating wall vessels for simulation of micro-g, and assessed for expansion, cell cycle activation, apoptosis, and hematopoietic potential. While CD34+ cells cultured in normal gravity (1-g) proliferated up to threefold by day 4-6, cells cultured in micro-g did not increase in number. As a possible explanation for this, cells cultured in simulated micro-g were found to exit G0/G1 phase of cell cycle at a slower rate than 1-g controls. When assayed for primitive hematopoietic potential in secondary conventional 1-g long-term cultures, cells from initial micro-g cultures produced greater numbers of cells and progenitors, and for a longer period of time, than cultures initiated with 1-g control cells. Similar low levels of apoptosis and adhesion molecule phenotype in micro-g and 1-g-cultured cells suggested similar growth patterns in the two settings. These data begin to elucidate the effects of micro-g on proliferation of human hematopoietic cells and may be potentially beneficial to the fields of stem cell biology and somatic gene therapy.

Antigens, CD34↗

Simulated microgravity [bed rest] has little influence on taste, odor or trigeminal sensitivity.

Anecdotal evidence suggests that astronauts' perceptions of foods in space flight may differ from their perceptions of the same foods on Earth. Fluid shifts toward the head experienced in space may alter the astronauts' sensitivity to odors and tastes, producing altered perceptions. Our objective was to determine whether head-down bed rest, which produces similar fluid shifts, would produce changes in sensitivity to taste, odor or trigeminal sensations. Six subjects were rested three times prior to bed rest, three times during bed rest and two times after bed rest to determine their threshold sensitivity to the odors isoamylbutyrate and menthone, the tastants sucrose, sodium chloride, citric acid, quinine and monosodium glutamate, and to capsaicin. Thresholds were measured using a modified staircase procedure. Self-reported congestion was also recorded at each test time. Thresholds for monosodium glutamate where slightly higher during bed rest. None of the other thresholds were altered by bed rest.

Adult↗

Plant growth processes in Arabidopsis under microgravity conditions simulated by a clinostat.

The life cycle of Arabidopsis plants was examined by growing them on a horizontal clinostat. Seeds on agar media were allowed to germinate and seedlings were grown under a simulated microgravity on a horizontal clinostat. Clinorotation (3 rpm) did not appear to interfere with germination of plant seeds and development of cotyledons and leaves. Stress relaxation parameters of the cell wall, the minimum relaxation time and the relaxation rate did not appear to be affected by clinostat rotation. On the other hand, the length of inflorescences was reduced to 61-62% by clinostat rotation. Rotation was found to inhibit the polar transport of auxin, although inflorescence growth and auxin transport were not completely inhibited. From these facts, it is possible that the life cycle in Arabidopsis plants could be accomplished in space, although growth phenomena involving auxin transport and its action may be disturbed. Plants may have a capacity to grow in space and we may be able to cultivate crops in space.

Arabidopsis↗

Head-out immersion in the non-human primate: a model of cardiovascular deconditioning during microgravity.

BACKGROUND: Orthostatic intolerance is a common complication associated with spaceflight. It has been speculated that this is due to changes in blood volume and alterations in cardiovascular reflexes. The objective of the current study was to develop a model that would allow us to study the cardiovascular system and the regulation of blood volume during short-term microgravity exposure in the primate with the intent of eventually being able to elucidate those factors responsible for the orthostatic intolerance. HYPOTHESIS: Head-out water immersion in the conscious non-human primate simulates the cardiovascular and volume regulatory responses observed in astronauts during exposure to microgravity. METHODS: Four monkeys were chronically instrumented for measuring BP and heart rate and then conditioned to the primate restraint chair. They were then subjected to 72 h of head-out water immersion (two immersions in three monkeys and one immersion in the fourth) in order to simulate the cardiovascular and renal effects of the microgravity environment. RESULTS: During the immersion, there was an increase in arterial BP (ABP) and central venous pressure (CVP) and a reflex decrease in heart rate (HR). Urine flow (UV) increased and water intake decreased, producing a negative water balance. This was not associated with an alteration in food intake. CVP and UV decreased following de-immersion. There was also resetting of the arterial baroreflex control of HR. Significant tachycardia occurred after the immersion that was associated with a decrease in ABP. CONCLUSION: These results are similar to those observed in astronauts during and after spaceflight, suggesting that head-out water immersion of the non-human primate provides a good model for studying cardiovascular and renal adaptations to spaceflight.

Animals↗

Peripheral effector mechanism hypothesis of postflight cardiovascular dysfunction.

Studies on the mechanisms of cardiovascular dysfunction after space-flight are important to illustrate the cardiovascular effect of microgravity and develop appropriate multi-system countermeasures for future long-duration spaceflights. Over the past 10 yr, we have systematically studied the adaptational changes in structure and function of both the heart and vessels, using the tail-suspension rat model to simulate microgravity effects. Our results indicate that simulated microgravity induced atrophic changes and reduced contractility of the heart muscle, and upward- and downward-regulation in structure, function, and innervation state of vessels in the brain and hind body of the rat. In addition, more recent advances in relevant ground-based and space-flight studies from different laboratories have also been reviewed. Based on these studies, it has been speculated that, in addition to hypovolemia, the microgravity-induced adaptational changes in the structure and function of the two main effectors of the cardiovascular system, i.e., the arterial smooth muscle and the cardiac muscle, might be among the most important mechanisms responsible for postflight cardiovascular dysfunction and orthostatic intolerance. In this paper we will review the available evidence with comments.

Adaptation, Physiological↗

Physiological reactions of primates to 9-D immersion and head-down tilt.

Purpose of the investigation was to compare physiological reactions of primates (Macaca mulatta) to microgravity simulated by immersion and head-down tilt (HDT). In immersion experiments, primates in waterproof suits were put into motion-restraining chairs and immersed into water (t=35.4 degrees C) breast-deep for 9 days. In 9-d HDT experiments, prone primates were motor restrained in dedicated tilt beds at -5 degrees. It was found that the CNS functioning was significantly affected, the plasma volume reduced and the marrow erythropoietic function declined. Atrophy developed in leg muscles on a backdrop of iliopectineal spongy osteopenia. Loss in hydration, inhibition of erythropoietic hemopoiesis and iliopectineal spongy osteopenia were more pronounced following immersion than HDT.

Adaptation, Physiological↗

Cardiovascular effects of simulated zero-gravity in humans.

UNLABELLED: Head-down and head-up [correction of heat-up] tilted bedrest (5 degrees) and head out water immersion (HOWI) for 6 hr were compared. PARAMETERS: Cardiac output (rebreathing method), blood pressure (arm cuff), forearm blood flow (venous occlusion plethysmography), total peripheral (TPR), and forearm vascular (FVR) resistances, Hct, Hb, relative plasma volume (PV) changes, and plasma catecholamines (single-isotope assay). During HOWI there was as expected a decrement in TPR, FVR, Mean arterial pressure (MAP, from 100 to 80 mmHg), Hct, and PV, and--as a new finding--catecholamines, which were 30-50% lower compared with both +5 and -5 degrees bedrest. During head down tilt, MAP was elevated (to 100-110 mmHg) and catecholamines did not fall, while TPR and EVR slowly decreased over 6 hr. HOWI is a stronger stimulus than -5 degrees bedrest, probably because HOWI elevates central venous pressure more markedly emptying the peripheral veins, while bedrest permits a distension of veins, which induces an increase in sympathetic nervous activity.

Adaptation, Physiological↗

Changes in enzymes and potassium content of the neuromuscular systems of albino rats during prolonged exposure to simulated hypogravics.

With respect to neuromuscular function, aldosterone activity, enzymatic and potassium (K) metabolism of organ tissues were investigated during the stress and adaptation stabilized phases of hypodynamically stressed rats. During adaptation, muscle tissue enzymes, such as aldolase, showed no change until the 35th day. The decrease of succinic dehydrogenase (SDH) was evident at 7 days. Lactic dehydrogenase (LDH) and creatine phosphokinase (CPK) serum levels increased transiently on the 18th day; this implied the development of muscular atrophy. A decrease in the 42K uptake of muscle was found from the 18th day onward. In the brain, a progressive decrease of aldolase was observed. 42K uptake showed no change in the brain, but the K content increased at both 7 and 18 days of exposure. The increase of cholinesterase (ChE) was more remarkable in the brain than in muscle, although transient. We suggest that the brain plays an important part in the adaptation process, through increasing or maintaining the functions of the neuromuscular excitation system during the 7-18 days of hypodynamic exposure.

Adaptation, Physiological↗

Changes in the rheological properties of the cell wall of plant seedlings under simulated microgravity conditions.

In this review article, we discuss the changes in the mechanical properties of the primary cell wall of different organs of several species of plant seedlings grown under simulated microgravity conditions. We compared growth and tropistic responses of these organs growing under different microgravity conditions, namely: 1) 3-D clinostat (three-dimensional clinostat with two axes); 2) water-submergence for rice seedlings, and 3) for comparison, an accelerated gravity using a specifically designed centrifuge. We measured the minimum stress-relaxation time as the parameter representing the mechanical property of the cell wall. We also measured extensiblity, in mm/g. The 3-D clinostat condition disturbed the normal gravitropic response of organs but affected growth rate and mechanical properties of the cell wall very little. Water-submergence of rice seedlings caused an acceleration of coleoptile elongation in the dark and caused a marked change in the mechanical property of the cell wall. However, the additional gravity of 30-135 xg showed only a small effect on growth and the mechanical property of the cell wall.

Biomechanical Phenomena↗

Vasomotor sympathetic nerve responses to static handgrip after simulated microgravity.

During the isometric exercise, the autonomic nervous system has a major role in the regulation of arterial blood pressure, blood flow, and perfusion to the active skeletal muscle in humans. An increase in cardiac sympathetic nerve activity and a withdrawal of vagal tone cause elevations of heart rate, cardiac contractility and cardiac output. An increase in vasomotor sympathetic nerve activity causes a vascular constriction to redirect the oxygen transport to the contracting muscle. It has been reported that the autonomic and cardiovascular systems are commonly affected by the exposure to the real (i.e. spaceflight) and simulated microgravitational environment( i.e. 6 degrees head-down bed rest (HDBR)). The alteration in the autonomic system includes a reduced vagal tone, and an attenuated carotid-cardiac baroreflex function. It may also include an altered control of peripheral vessels, a reduced sympathetic vasoconstriction, an attenuated release of norepinephrine from sympathetic nerve terminal, an impaired myogenic contractility, an up- or down regulation of alpha-adrenergic receptors, and an attenuated multiple vasodilatation. Therefore, there seems to be a possibility that HDBR alters the sympathetic and cardiovascular responses to isometric exercise in humans, however, this possibility has not been examined efficiently. We compared the vasomotor sympathetic and cardiovascular responses to sustained handgrip (HG) until fatigue followed by post-exercise muscle ischemia (PEMI) after 14 days of HDBR, to those before HDBR, in 16 healthy males.

Adult↗

Effects of microgravity on c-fos gene expression in osteoblast-like MC3T3-E1 cells.

The paper summarizes the data on proliferation and gravity-related gene expression of osteoblasts that were obtained from an experiment conducted under simulated and real microgravity conditions. Simulated microgravity conditions obtained in a clinostat depress proliferation of both osteoblast-like MC3T3-E1 and HeLa carcinoma cells. This depression of proliferation occurs in a collagen gel culture in which the flow of culture medium by rotation may be reduced. Interestingly, MC3T3-E1 cells which are probably one of target cells to microgravity are more sensitive than the HeLa cells. Simulated microgravity inhibited the epidermal growth factor (EGF)-induced c-fos gene expression in the MC3T3-El cells. To examine in detail the effect of real microgravity on the EGF signal transduction cascade in osteoblasts, MC3T3-E1 cells were cultured in the Cell Culture Experiment Module of the sounding rocket TR-1A6. The EGF-induced c-fos expression in cells was depressed under short-term microgravity conditions in the sounding rocket, while the phosphorylation of mitogen-activated protein kinase (MAPK) was not affected compared with the controls grown on the ground. These results suggest that an action site of microgravity in the signal transduction pathway may be downstream of MAPK.

Animals↗

[Graviresponse in higher plants and its regulation in molecular bases: relevance to growth and development, and auxin polar transport in etiolated pea seedlings].

We review the graviresponse under true and simulated microgravity conditions on a clinostat in higher plants, and its regulation in molecular bases, especially on the aspect of auxin polar transport in etiolated pea (Pisum sativum L. cv. Alaska) seedlings which were the plant materials subjected to STS-95 space experiments. True and simulated microgravity conditions substantially affected growth and development in etiolated pea seedlings, especially the direction of growth of stems and roots, resulting in automorphosis. In etiolated pea seedlings grown in space, epicotyls were the most oriented toward the direction far from the cotyledons, and roots grew toward the aerial space of Plant Growth Chamber. Automorphosis observed in space were well simulated by a clinorotation on a 3-dimensional clinostat and also phenocopied by the application of auxin polar transport inhibitors of 2,3,5-triiodobenzoic acid, N-(1-naphtyl)phthalamic acid and 9-hydroxyfluorene-9-carboxylic acid. Judging from the results described above together with the fact that activities of auxin polar transport in epicotyls of etiolated pea seedlings grown in space substantially were reduced, auxin polar transport seems to be closely related to automorphosis. Strenuous efforts to learn in molecular levels how gravity contributes to the auxin polar transport in etiolated pea epicotyls resulted in successful identification of PsPIN2 and PsAUX1 genes located in plasma membrane which products are considered to be putative efflux and influx carriers of auxin, respectively. Based on the results of expression of PsPIN2 and PsAUX1 genes under various gravistimulations, a possible role of PsPIN2 and PsAUX1 genes for auxin polar transport in etiolated pea seedlings will be discussed.

Biological Transport↗

Regional blood flow in conscious rats after head-down suspension.

Exposure to microgravity in humans causes cardiovascular deconditioning affecting blood pressure, heart rate and vascular responsiveness. This study investigated cardiac output, arterial blood pressure and regional blood flows [radioactive microspheres: 57Co, 15.5 (SEM 0.1) microm in diameter] in conscious and freely moving rats subjected to 14 days of simulated microgravity (head-down suspension, HDS) in male Wistar rats: control (horizontally attached, n = 7), suspended for 14 days (n = 8) and suspended/allowed to recover for 10 min (R10min, n = 5) or 24 h (n = 9). Compared to the control group, 14 days of HDS resulted in reduced total peripheral resistance (37%); an increased cardiac index (65%) was associated with no significant change in the mean arterial pressure BPa. There were elevated brain (63%), visceral (> 20%), hindlimb (> 80%) and forelimb (> 215%) muscle blood flows. In the R10min group, the BPa decreased (18%) and the regional blood flows returned to control values. Within 24 h the BPa as well as cardiac index and total peripheral resistance were restored. In conclusion, 14 days of HDS engendered local circulatory changes resulting in transient blood pressure instability during recovery.

Animals↗

Expression of PIN and AUX1 genes encoding putative carrier proteins for auxin polar transport in etiolated pea epicotyls [correction of epicotyles] under simulated microgravity conditions on a three-dimensional clinostat.

Etiolated pea (Pisum sativum L. cv. Alaska) seedlings grown under simulated microgravity conditions on a 3-dimensional clinostat showed automorphosis-like growth and development similar to that observed in true microgravity conditions in space. Application of inhibitors of auxin polar transport phenocopied automorphosis-like growth on 1 g conditions, suggesting that automorophosis is closely related to auxin polar transport. Strenuous efforts to know the relationships between automorphosis and auxin polar transport in pea seedlings at molecular bases resulted in successful identification of PsPIN2 and PsAUX1 encoding putative auxin efflux and influx carrier protein, respectively. Significantly high levels in homology were found on nucleotide and deduced amino acid sequences among PsPIN2, PsPIN1 and AtPINs, and between PsAUX1 and AtAUX1. Expression of PsPIN1 and PsAUX1 genes in etiolated pea seedlings grown on the clinostat were substantially affected, but that of PsPIN2 was not. Roles of these genes in auxin polar transport and automorphosis of etiolated pea seedlings are also described.

Biological Transport↗

Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture system.

The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.

Animals↗