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PubMed1996

Clinorotation affects morphology and ethylene production in soybean seedlings.

The microgravity environment of spaceflight influences growth, morphology and metabolism in etiolated germinating soybean. To determine if clinorotation will similarly impact these processes, we conducted ground-based studies in conjunction with two space experiment opportunities. Soybean (Glycine max [L.] Merr.) seeds were planted within BRIC (Biological Research In Canister) canisters and grown for seven days at 20 degrees C under clinorotation (1 rpm) conditions or in a stationary upright mode. Gas samples were taken daily and plants were harvested after seven days for measurement of growth and morphology. Compared to the stationary upright controls, plants exposed to clinorotation exhibited increased root length (125% greater) and fresh weight (42% greater), whereas shoot length and fresh weight decreased by 33% and 16% respectively. Plants grown under clinorotation produced twice as much ethylene as the stationary controls. Seedlings treated with triiodo benzoic acid (TIBA), an auxin transport inhibitor, under clinorotation produced 50% less ethylene than the untreated control subjected to the same gravity treatment, whereas a treatment with 2,4-D increased ethylene by five-fold in the clinorotated plants. These data suggest that slow clinorotation influences biomass partitioning and ethylene production in etiolated soybean plants.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1999

Effect of horizontal clinorotation on the root system development and on lipid breakdown in rapeseed (Brassica napus) seedlings.

Seedlings of Brassica napus were cultivated on a slowly rotating clinostat (1 rpm) or in the vertical control for 5 d. The root growth, the cotyledonary reserves and the transport of 14C-labeled sucrose from cotyledons to root system were studied in both cultural conditions. The biomass (fresh weight) of the root system was 35% higher in the horizontally clinorotated seedlings than in the controls. This increase was correlated with a greater degradation of reserve lipids and faster accumulation of sucrose in the cotyledons. The activity of isocitrate lyase, one of the two enzymes necessary to conversion of lipids into glucids, was also greater in the cotyledons of clinorotated seedlings. The labeling distribution of 14C in the cotyledons, the hypocotyl and the root system after 30, 60 and 120 min of application of 14C-labeled sucrose on the cotyledons showed higher translocation of the cotyledonary sucrose to the root system of clinorotated seedlings. In addition, we studied the effects of clinorotation on the biomass of the excised root system (of 10 d old seedlings) cultivated in a medium containing 1% sucrose. The horizontally clinorotated root system grew more than that of the controls. These results showed that the horizontal clinorotation acted on the root system growth and provoked a higher sucrose translocation from source to sink, i.e. from cotyledons to root system.

Biological Transport↗
PubMed2002

Clinostat rotation induces apoptosis in luteal cells of the pregnant rat.

Recent studies have shown that microgravity induces changes at the cellular level, including apoptosis. However, it is unknown whether microgravity affects luteal cell function. This study was performed to assess whether microgravity conditions generated by clinostat rotation induce apoptosis and affect steroidogenesis by luteal cells. Luteal cells isolated from the corpora lutea of Day 8 pregnant rats were placed in equal numbers in slide flasks (chamber slides). One slide flask was placed in the clinostat and the other served as a stationary control. At 48 h in the clinostat, whereas the levels of progesterone and total cellular protein decreased, the number of shrunken cells increased. To determine whether apoptosis occurred in shrunken cells, Comet and TUNEL assays were performed. At 48 h, the percentage of apoptotic cells in the clinostat increased compared with that in the control. To investigate how the microgravity conditions induce apoptosis, the active mitochondria in luteal cells were detected with JC-1 dye. Cells in the control consisted of many active mitochondria, which were evenly distributed throughout the cell. In contrast, cells in the clinostat displayed fewer active mitochondria, which were distributed either to the outer edge of the cell or around the nucleus. These results suggest that mitochondrial dysfunction induced by clinostat rotation could lead to apoptosis in luteal cells and suppression of progesterone production.

Animals↗
PubMed2003

Loss of T cell precursors after spaceflight and exposure to vector-averaged gravity.

Using fetal thymus organ culture (FTOC), we examined the effects of spaceflight and vector-averaged gravity on T cell development. Under both conditions, the development of T cells was significantly attenuated. Exposure to spaceflight for 16 days resulted in a loss of precursors for CD4+, CD8+, and CD4+CD8+ T cells in a rat/mouse xenogeneic co-culture. A significant decrease in the same precursor cells, as well as a decrease in CD4-CD8- T cell precursors, was also observed in a murine C57BL/6 FTOC after rotation in a clinostat to produce a vector-averaged microgravity-like environment. The block in T cell development appeared to occur between the pre-T cell and CD4+CD8+ T cell stage. These data indicate that gravity plays a decisive role in the development of T cells.

Animals↗
PubMed2005

Functional recovery of peripheral blood mononuclear cells in modeled microgravity.

Immune function is suppressed in space flight, demonstrated by reduced mitogen-stimulated proliferation of postflight astronaut peripheral blood mononuclear cells (PBMCs). While flight studies are limited, the development of rotating wall vessel (RWV) bioreactors, such as the high aspect ratio vessel (HARV), has facilitated ground-based studies of the effects of modeled microgravity (MMG) on cell-mediated immunity. Astronauts regain immune function 3 days postflight, but this recovery has not yet been demonstrated following MMG. MMG eliminated phytohemagglutinin (PHA)-stimulated proliferation of PBMCs. Upon removal from HARV, full recovery was gradually achieved over a 72 h period, in agreement with postflight studies of astronauts. Recovery from MMG delayed, but did not reduce, the maximal proliferative response compared with PHA-activated stationary cultures. Likewise, peak expression of T cell surface activation markers CD69 and CD25 was delayed upon stimulation following exposure to MMG. MMG and recovery from MMG differentially affected the detection of IL-2 and IFN in supernatants. Further development of this model of immune recovery is important for investigating the mechanisms of immune suppression and recovery in space flight, as well as possible countermeasures to prevent immunosuppression or enhance recovery. Given the analogous immune suppression observed in microgravity, MMG, and aging, further investigation may also lead to advances in anti-aging medicine.

Adolescent↗
PubMed2006

Airway management with endotracheal tube versus Combitube during parabolic flights.

BACKGROUND: Training of National Aeronautics and Space Administration space shuttle astronauts revealed difficult airway management with endotracheal tubes (ETTs) under microgravity conditions. The authors performed a randomized comparative study of ETT and Combitube (ETC; Tyco Healthcare, Pleasanton, CA). The aim of the study was to evaluate ease, time of insertion, and success rates during normogravity and parabolic flights using mannequins. METHODS: After normogravity experiments, four flyers performed intubation on a mannequin during the flights. Sixty-two intubation attempts were performed using the ETC (normogravity, 29; microgravity, 33), and 58 intubation attempts were performed using the ETT (each 29 attempts, both conditions). Time to completion of the intubation procedure, success rate, and ease of insertion were recorded. RESULTS: The ETC performed equally well between normogravity (median, 18 s; range, 17-25 s) and microgravity (median, 18.5 s; range, 17-28 s), whereas the ETT performed significantly slower under microgravity (median, 20 s; range, 17-27 s) as compared with normogravity (median, 18 s; range, 16-22 s; P = 0.019). One hundred nine of 120 (90%) were successful. The ETT and ETC were comparable with respect to successful intubations, under normogravity or microgravity, respectively. CONCLUSIONS: Both the ETC and ETT perform comparably well. Slight differences could be found with respect to time of insertion in favor of the ETC. Because this is the first experiment using the ETC on the KC-135, it is shown that there is enough time to perform the insertion procedure. Because the ETC airway requires less training and is easier to insert than an ETT, it is recommended for further study as an alternative airway to what is currently on the shuttle.

Electronics↗
PubMed1996

Regional blood flow in microgravity: adaptation and deconditioning.

The objectives were to evaluate cardiac and peripheral changes induced by microgravity with and without countermeasures (CM), to assess the peripheral response to orthostatic tests (tilt, LBNP). Inflight or HDT, we used echography and Doppler to assess the left heart function and the peripheral arteries. We studied the cardiovascular system during 1) 21-d and 25-d spaceflights without CM, 2) 14.d spaceflight with "bracelets" CM, 3) 28-d HDT with and without LBNP, and 4) 30-d HDT with and without Exercise+LBNP. Similar peripheral circulation changes were noticed in both astronauts and HDT subjects without CM. There was a decrease in renal, cerebral, and femoral vascular resistances and maintenance of cerebral flow at rest, and a lack of increase in lower limb vascular resistance and abnormal flow redistribution during orthostatic tests. Conversely, with CM at rest, cerebral and renal vascular resistances stayed elevated and femoral resistance decreased, but less than without countermeasures. Lower limb vascular resistance increased normally, peripheral flows were adequately redistributed during orthostatic tests, and no orthostatic intolerance was observed. This confirms the efficiency of countermeasures (LBNP, exercise, cuffs) in preserving the vasomotor tone in most peripheral areas at rest and reducing the development of orthostatic intolerance.

Adaptation, Physiological↗
PubMed2002

Microgravity culture condition reduces immunogenicity and improves function of pancreatic islets1.

BACKGROUND: The failure of pancreatic islet allotransplants observed in almost all clinical attempts is related to poor initial islet function and allograft rejection. To remedy these problems we cultured islets in microgravity conditions to improve their function and to reduce their immunogenicity. METHODS: Fresh mouse islets or mouse islets cultured in stationary dishes or microgravity bioreactors were transplanted to streptozotocin-induced diabetic mouse recipients. RESULTS: Both allogeneic dish- or bioreactor-cultured islets survived more than 100 days compared with fresh allogeneic islets, which were rejected in less than 15 days. Islet titration studies revealed that 250 fresh or dish-cultured, but only 30 to 120 bioreactor-cultured, islets were necessary to produce euglycemia. Furthermore, glucose tolerance tests showed that bioreactor-cultured islets functioned better compared with fresh and dish-cultured islets on day 30 postgrafting. Immunostaining and transmission electron microscopy (TEM) analyses showed the gradual disappearance of dendritic cells in cultured islets compared with fresh islets. TEM revealed that the ultrastructure of islets from bioreactor, but not dish, appeared healthy and closely resembled fresh islets. Interestingly, TEM and scanning electron microscopy showed that only bioreactor-cultured islets developed unique and multiple nutritional channels between arrays of islet cells. TEM with colloidal lanthanum tracer revealed that only bioreactor islet cell cultures were devoid of tight junctional complexes, which may facilitate channel formation. CONCLUSION: Microgravity condition decreases immunogenicity and significantly improves the function of secretory cells.

Animals↗
PubMedDate not supplied

Space flight and the skeleton: lessons for the earthbound.

Loss of bone during extended space flight has long been a concern that could limit the ability of humans to explore the universe. Surprisingly, the available data do not support the concept that weightlessness leads inexorably to a depleted skeleton unable to withstand the stress of a return to a 1-g environment. Nevertheless, some bone loss does occur, especially in those bones most stressed by gravity prior to flight, which provides confirmation of the proposal formulated over a century ago by Julius Wolff that mechanical stress determines the form and function of bone. Although the phenomenon of bone loss with skeletal unloading, whether by space flight or immobilization or just taking a load off your feet (literally) is well established, the mechanisms by which bone senses load and adjusts to it are not so clear. What actually is the stimulus, and what are the sensors? What are the target cells? How do the sensors communicate the message into the cells, and by what pathways do the cells respond? What is the role of endocrine, factors vs. paracrine or autocrine factors in mediating or modulating the response? None of these questions has been answered with certainty, but, as will become apparent in this review, we have some clues directing us to the answers. Although the focus of this review concerns space flight, it seems highly likely that the mechanisms mediating the transmission of mechanical load to changes in bone formation and resorption apply equally well to all forms of disuse osteoporosis and are likely to be the same mechanisms affected by other etiologies of osteoporosis.

Animals↗
PubMed2003

Microgravity as a model of ageing.

PURPOSE OF REVIEW: Longevity with good health and long-term survival in space are two of the many challenges that scientists face in the twenty-first century. Ageing and life in space are both associated with undesirable effects on normal physiological processes. This review will outline how the endocrine, metabolic, immune and musculoskeletal systems are affected by microgravity and ageing, drawing analogies between the observed changes in an attempt to highlight common mechanisms. RECENT FINDINGS: Mild hypothyroidism, increased stress hormones (mainly catecholamines), decreased sex steroids, insulin resistance, impaired anabolic response to food intake, anorexia, altered mitochondrial function and systemic inflammatory response are common features of both ageing and microgravity. Both conditions lead to progressive bone and muscle atrophy, compromising mobility and the ability to perform essential daily tasks. In skeletal muscle, both ageing and space flight lead to weakness from whole muscle to single fibre level, accompanied by marked alterations in muscle architecture and in tendon mechanical properties. SUMMARY: What makes microgravity an interesting and unique tool for gerontologists is that many space-related physiological changes resemble those observed during ageing, but are more or less quickly restored after re-entry, thus allowing the biology of ageing to be investigated both ways, not only during its development but also during recovery.

Adaptation, Physiological↗
PubMed1990

Circumnutations of sunflower hypocotyls in satellite orbit.

The principal objective of the research reported here was to determine whether a plant's periodic growth oscillations, called circumnutations, would persist in the absence of a significant gravitational or inertial force. The definitive experiment was made possible by access to the condition of protracted near weightlessness in an earth satellite. The experiment, performed during the first flight of Spacelab on the National Aeronautics and Space Administration shuttle, Columbia, in November and December, 1983, tested a biophysical model, proposed in 1967, that might account for circumnutation as a gravity-dependent growth response. However, circumnutations were observed in microgravity. They continued for many hours without stimulation by a significant g-force. Therefore, neither a gravitational nor an inertial g-force was an absolute requirement for initiation [correction of initation] or continuation of circumnutation. On average, circumnutation was significantly more vigorous in satellite orbit than on earth-based clinostats. Therefore, at least for sunflower (Helianthus annuus L.) circumnutation, clinostatting is not the functional equivalent of weightlessness.

Acceleration↗
PubMed2004

Mechanotransduction in root gravity sensing cells.

The analysis of the dose-response curve of the gravitropic reaction of lentil seedling roots has shown that these organs are more sensitive when they have been grown in microgravity than when they have been grown on a 1 g centrifuge in space before gravistimulation. This difference of gravisensitivity is not due to the volume or the density of starch grains of statoliths, which are about the same in both conditions (1 g or microgravity). However, the distribution of statoliths within the statocyte may be responsible for this differential sensitivity, since the dispersion of these organelles is greater in microgravity than in 1 g. When lentil roots grown in microgravity or in 1 g are stimulated at 0.93 g for 22 min, the amyloplasts sediment following two different trajectories. They move from the proximal half of the statocytes toward the lower longitudinal wall in the microgravity grown sample and from the distal half toward the longitudinal wall in the 1 g grown sample. At the end of the stimulation, they reach a similar position within the statocytes. If the roots of both samples are left in microgravity for 3 h, the amyloplasts move toward the cell centre in a direction that makes an average angle of 40 degrees with respect to the lower longitudinal wall. The actin filaments, which are responsible for this movement, may have an overall orientation of 40 degrees with respect to this wall. Thus, when roots grown in microgravity are stimulated on the minicentrifuge the amyloplasts slide on the actin filaments, whereas they move perpendicular to them in 1 g grown roots. Our results suggest that greater sensitivity of seedling roots grown in microgravity should be due to greater dispersion of statoliths, to better contacts between statoliths and the actin network and to greater number of activated mechanoreceptors. One can hypothesize that stretch activated ion channels (SACs) located in the plasma membrane are responsible for the transduction of gravistimulus. These SACs may be connected together by elements of the cytoskeleton lining the plasma membrane and to the actin filaments. They could be stimulated by the action of statoliths on the actin network and/or on these elements of the cytoskeleton which link the mechanoreceptors (SACs).

Actins↗
PubMed2005

Effects of 84-days of bedrest and resistance training on single muscle fibre myosin heavy chain distribution in human vastus lateralis and soleus muscles.

AIM: This investigation determined the effects of 84 days of bedrest on the composition of myosin heavy chain (MHC) in single skeletal muscle fibres with and without a resistance-training countermeasure programme. METHODS: Muscle biopsies were obtained from the m. vastus lateralis (VL) and m. soleus (SOL) before and after 84 days of bedrest. While control (BR) subjects (VL n = 9; SOL n = 3) refrained from exercise, BRE subjects (VL n = 8; SOL n = 3) performed knee extensor and plantar flexor resistance exercise every third day. Approximately 110 fibres per sample were analysed for MHC composition using SDS-PAGE. RESULTS: BR-VL had 16 and 14% decreases (P < 0.05) in MHC I and IIa fibres, respectively. There were 10% increases (P < 0.05) in MHC I/IIa, IIa/IIx, I/IIa/IIx, and a approximately 30% increase (P < 0.05) in total hybrid fibres. BRE-VL showed a 15% reduction (P < 0.05) in MHC I fibres, no change in MHC IIa fibres, and a 13% increase (P < 0.05) in total hybrids. BR-SOL had a 19% decrease (P < 0.05) in MHC I fibres with a 22% increase in total hybrids. BRE-SOL showed no change in MHC composition across all fibre types. CONCLUSION: These data suggest that the exercise countermeasures programme prevented MHC shifts in the SOL and mitigated MHC shifts in the VL. Furthermore, in the VL it appears that the resistance training programme employed in this investigation during bedrest, emphasized the use of MHC IIa phenotype muscle fibres.

Adult↗
PubMed2005

Morin sulphates/glucuronides enhance macrophage function in microgravity culture system.

BACKGROUND: The immune system changes significantly in astronauts during and after space flight. Although the mechanism has not been defined, it is reasonable to begin developing effective countermeasures to the physiological consequences of spaceflight, especially immunosuppression. Many studies have been published about the effect of flavonoids on immune modulation. Thus, the aim of this study was to develop whether flavonoids could be the effective countermeasures to the immunosuppression caused by microgravity. MATERIALS AND METHODS: We used a rotating wall vessel 3D (three-dimensional) culture system which recreates some of the culture conditions that occur during microgravity to study the effects of microgravity on the function of macrophages and assess the modulating effects of flavonoids on microgravity-induced macrophage dysfunction. RESULTS: We demonstrated 65% and 80% reduction in mitogen-induced nitric oxide and cytokine production of 3D-cultured macrophages, compared to conventional two-dimensional (2D)-cultured cells. Moreover, the microgravity-induced macrophage dysfunction was not restored by transferring cells from 3D to 2D culture. However, the addition of morin sulphates/glucuronides in 3D culture compensated for the loss of macrophage function. CONCLUSION: The result presented here suggests for the first time that an immune-modulatory strategy using flavonoid supplements such as morin would benefit the health of astronauts.

Animals↗
PubMed2006

The molecular chaperone HSP47 rapidly senses gravitational changes in myoblasts.

Skeletal muscle unloading induced by spaceflight or bed rest leads to muscle atrophy. It is unclear how muscle atrophy is caused and how muscles respond to microgravity. We addressed the response of collagen and its chaperone system to gravitational forces. We show here that expression of HSP47, a collagen-specific molecular chaperone, responds to gravitational changes, including microgravity and hypergravity in vitro and in vivo. By using the method hindlimb suspension of rats, which mimics microgravity conditions, we demonstrated that the expression of Hsp47 mRNA decreased within 1 day and the mRNA levels of collagen types I and IV were subsequently reduced. In contrast, hypergravity stimulated HSP47 expression. HSP47 and collagen types I and IV were localized intracellularly in the endoplasmic reticulum and/or Golgi apparatus of myoblasts, as expected. Intriguingly, Hsp47 mRNA levels in cultured myoblasts increased significantly with hypergravity treatment at 40G for 2 h, and decreased with microgravity treatment at almost 0G for 1-2 h. Collagen mRNA levels were also altered, although changes were slower and less pronounced compared with those for HSP47. The gravity-regulated HSP47 may play a role in the maintenance of the extracellular matrix by modulating collagen production at the primary stage of adaptation.

Animals↗
PubMed1994

Starch metabolism in germinating soybean cotyledons is sensitive to clinorotation and centrifugation.

Soybean (Glycine max [L.] Merr. cv. McCall) seedlings germinated and grew for 6d under the altered gravity conditions of horizontal clinorotation and centrifugation. Both of these conditions resulted in decreased growth relative to the control (vertically rotated) plants. Starch concentration in the cotyledons was lower in the clinorotated plants and was higher in the centrifuged plants compared to the controls. The opposite relationship was noted for total lipid concentration. Of the six starch metabolic enzyme activities measured, only ADP glucose pyrophosphorylase was affected by the gravity treatments; being lower in the cotyledons of the horizontally rotated plants and higher in the cotyledons of the centrifuged plants relative to the control values.

Centrifugation↗