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PubMed1998

Functional weightlessness during clinorotation of cell suspensions.

A clinostat is a device often used in gravitational biology studies. Selecting an appropriate speed of rotation, however, is a frequently debated topic, particularly for suspended cells. In an attempt to define the necessary criteria for determining an acceptable revolution speed, the primary forces governing particle behavior during clinorotation--gravity, diffusion and centrifugation--were mathematically assessed. In support of the theoretical exercise, bacterial growth experiments indicated that results obtained using a clinostat followed trends resembling previous space flight results. It is suspected that this is due, in part at least, to similarly altered external transport processes in each environment.

Bacterial Physiological Phenomena↗
PubMed1998

Study of adaptation to altered gravity through systems analysis of motor control.

Maintenance of posture and production of functional, coordinated movement demand integration of sensory feedback with spinal and supra-spinal circuitry to produce adaptive motor control in altered gravity (G). To investigate neuroplastic processes leading to optimal performance in altered G we have studied motor control in adult rats using a battery of motor function tests following chronic exposure to various treatments (hyper-G, hindlimb suspension, chemical distruction of hair cells, space flight). These treatments differentially affect muscle fibers, vestibular receptors, and behavioral compensations and, in consequence, differentially disrupt air righting, swimming, posture and gait. The time-course of recovery from these disruptions varies depending on the function tested and the duration and type of treatment. These studies, with others (e.g., D'Amelio et al. in this volume), indicate that adaptation to altered gravity involves alterations in multiple sensory-motor systems that change at different rates. We propose that the use of parallel studies under different altered G conditions will most efficiently lead to an understanding of the modifications in central (neural) and peripheral (sensory and neuromuscular) systems that underlie sensory-motor adaptation in active, intact individuals.

Adaptation, Physiological↗
PubMed1999

Agravitropic mutant for the study of hydrotropism in seedling roots.

Roots have been shown to respond to a moisture gradient by positive hydrotropism. Agravitropic mutant plants are useful for the study of the hydrotropism in roots because on Earth hydrotropism is obviously altered by the gravity response in the roots of normally gravitropic plants. The roots are able to sense water potential gradient as small as 0.5 MPa mm(-1). The root cap includes the sensing apparatus that causes a differential growth at the elongation region of roots. A gradient in apoplastic calcium and calcium influx through plasmamembrane in the root cap is somehow involved in the signal transduction mechanism in hydrotropism, which may cause a differential change in cell wall extensibility at the elongation region. We have isolated an endoxy loglucan transferase (EXGT) gene that is strongly expressed in pea roots and appears to be involved in the differential growth in hydrotropically responding roots. Thus, it is now possible to study hydrotropism in roots by comparing with or separate from gravitropism. These results also imply that microgravity conditions in space are useful for the study of hydrotropism and its interaction with gravitropism.

Calcium Channels↗
PubMed1999

Space flight, microgravity, stress, and immune responses.

Exposure of animals and humans to space flight conditions has resulted in numerous alterations in immunological parameters. Decreases in lymphocyte blastogenesis, cytokine production, and natural killer cell activity have all been reported after space flight. Alterations in leukocyte subset distribution have also been reported after flight of humans and animals in space. The relative contribution of microgravity conditions and stress to the observed results has not been established. Antiorthostatic, hypokinetic, hypodynamic, suspension of rodents and chronic head-down tilt bed-rest of humans have been used to model effects of microgravity on immune responses. After use of these models, some effects of space flight on immune responses, such as decreases in cytokine function, were observed, but others, such as alterations in leukocyte subset distribution, were not observed. These results suggest that stresses that occur during space flight could combine with microgravity conditions in inducing the changes seen in immune responses after space flight. The biological/biomedical significance of space flight induced changes in immune parameters remains to be established. Grant Numbers: NCC2-859, NAG2-933.

Aerospace Medicine↗
PubMed2001

Time course of recovery of the somatosensory map following hindpaw sensory deprivation in the rat.

Hindlimb sensory deprivation is known to induce a decrease in the cortical representation of hindpaw, and an increase in the size of the cutaneous receptive fields. The aim of the present study was to determine (i) the time-course of recovery when the rat retrieves a normal use of its limbs after a 14-day period of sensory disruption and (ii) whether a 1-day period of sensory deprivation is sufficient to induce a plasticity. Our results indicate that the remodelling of the cortical map was not observed after 1 day of sensory deprivation. On the other hand, the recovery was achieved after 6 h. These findings suggest that a procedure reducing sensory function resulted in reversible changes in the somatosensory cortex. The recovery was more rapid than the induction of plasticity.

Animals↗
PubMed2001

Response of renal sympathetic nerve activity to parabolic flight-induced gravitational change in conscious rats.

The renal sympathetic nerve activity (RNA) response to gravitational changes induced by parabolic flight was examined in chronically instrumented conscious rats. Two types of RNA responses were found. In six out of 12 rats, the RNA did not respond during the 2 G period, but immediately fell to background levels on entry into microgravity (microG), then recovered to the 1 G control level during continued microG (shutdown obvious group). In the other six rats, the RNA increased to 158+/-13% at the end of the 2 G period, increased further to 195+/-22% on entry into microG, then gradually recovered to that seen at 1 G (shutdown obscure group). The mean arterial pressure in the shutdown obvious group was significantly higher and the heart rate tended to be higher than in the shutdown obscure group, suggesting that the baseline sympathetic tone in the shutdown obvious group was higher than in the shutdown obscure group. These results suggest that the RNA response to parabolic flight might be affected by the baseline sympathetic tone.

Animals↗
PubMed1999

A review of the effects of microgravity and of hypergravity on aging and longevity.

This article reviews the literature dealing with the effects of acute or chronic exposure to microgravity (spacecrafts) or hypergravity (centrifuge) on longevity and the aging process. Even if space flights are now common, the effects of these two kinds of environment on aging are still poorly documented, particularly in mammals. However, there is a growing interest for the use of the fruit fly Drosophila melanogaster, and this species may be now considered as a model organism in gravitational biology studies dealing with aging.

Aging↗
PubMed2004

Periodic structures and diurnal variation in blood pressure and heart rate in relation to microgravity on space station MIR.

Four Russian crew members were studied on space station MIR, and blood pressure (BP) and heart rate (HR) data were continuously collected. BP and HR data were collected on earth 1 day before orbital flight to the space station, then at weeks 8, 16 and 24 during space flight, and again 1 or 2 days after returning to earth. Time serial data for BP and HR were analyzed by spectral analysis with the MemCalc system (Suwa Trust, Sapporo, Japan). Periodic structures of diurnal variation in systolic blood pressure (SBP), diastolic blood pressure (DBP) and HR were compared at 24-hour, 12-hour and 8-hour intervals, these being determined as the main periodic components for the assessment of BP and HR variability. The 24-h mean levels of SBP and HR during space flight were unchanged. Waking SBP was not different from pre-flight values. During sleep, in-flight changes in HR did not differ from pre-flight values. SBP during sleep in orbit increased to over pre-flight values. Waking DBP was reduced during flight. The SBP and HR phases over a 24-hour cycle were shortened with a more pronounced shortening in weeks 8 and 16 compared with pre-flight values, and at week 24 recovered to preflight values. The 12, 8-hour-cycle remained unchanged, and were similar to pre-flight values. At the space station, the astronauts' mission was carried out under strict control of sleeping and waking hours; therefore, their 24-hour schedule is an artificially constructed situation. Main periodicity structures were maintained by strict control of lifestyle during long-term space flight. The conclusions reached were as follows: 1) SBP levels during sleep in a space environment increased compared with those on earth; 2) the periodicity phase of BP and HR shifted toward to 24-hour cycle as a result of long-term space flight, even though these periods shortened after a few months compared with pre-space flight values.

Adult↗
PubMed2002

Anorexia in space and possible etiologies: an overview.

Space travelers experience a flight duration-dependent loss in weight and body mass while in a microgravity environment, despite the absence of increased energy expenditure. Anorexia in space can lead to in-flight caloric deficits of 1330 kcal per 70 kg astronaut per day in the presence of abundant food and has a critical effect on endurance and performance. Microgravity, alterations in the light-and-dark cycle, and exposure to radiation energy are the environmental stresses believed to influence appetite, food intake, and gastrointestinal function during space flight. Review of data and recent studies in rodents during microgravity showed a release of stress hormones and complex neuroendocrine and physiologic changes involving the modulation of hypothalamic activity, food intake-related hormones, and cytokines. The shift of dietary preference to carbohydrates, which occurs in astronauts, denotes a stress physiologic response and augments free-plasma tryptophan concentration in the brain, the precursor of the potent anorexic agent, serotonin (5-HT). Alterations of other neuroendocrine mediators, including corticotropin-releasing factor (CRF), coordinate the stress response, leading to a decrease in appetite and gastrointestinal function. Our laboratories used the antiorthostatic tail-suspension technique to successfully mimic some of these anorexia-related stress responses and to directly demonstrate the role of 5-HT in microgravity-related decreased food intake and delayed gastric emptying. Further rodent studies from our laboratories demonstrated the adverse effect of altered dark-and-light cycles on food intake and body weight. Radiation energy, through its documented effects on appetite, probably contributes to the decreased caloric intake by astronauts. Modulation of hypothalamic activity, 5-HT, and CRF play a critical role in anorexia related to microgravity and circadian rhythm alterations. Specific gene knockout mice (e.g., 5-HT or CRF and their respective receptors) may prove fruitful in defining the pathways by which anorexia in space occurs. An understanding of these pathophysiologic problems as they relate to appetite, food intake, gastric emptying and gastrointestinal function, sufficiently to derive successful practical solutions, may lead to a quantitative enhancement of physiologic well-being and performance status, serving as a productive countermeasure in space.

Adaptation, Physiological↗
PubMed2003

Increased levels of UV-induced protease activity in human UVAP-1 cells exposed to gravity-changing stress: involvement of E-64-sensitive proteases in suppression of UV mutagenicity.

Under the 1G condition, the increase in antipain-sensitive protease activity promptly after UV (mainly 254 nm wavelength) irradiation in cultured human cells is detected and found to be one of the intriguing events involved in suppression of cell mutability. It was found that two cell lines, RSa and its variant UVAP-1 cells are applicable; the former is hypermutable and not susceptible to protease activation, while the latter is hypomutable and susceptible. In the present study it was investigated whether the increase in protease activity by UV irradiation is also observed in hypomutable human UVAP-1 cells exposed to gravity-changing stress and whether the increase is involved in suppression of UV mutagenicity. Exposure of human UVAP-1 cells to gravity-changing stress such as free-fall and parabolic flight prior to UV irradiation resulted in a pronounced increase in protease activity, but not to hypergravity conditions (2 and 10G) prior to UV irradiation. To characterize the proteases, components of lysates from the cells exposed to free-fall prior to UV irradiation were fractionated by high performance liquid chromatography, indicating two separate fractions with highly increased levels of E-64-sensitive protease activity. In the cells treated with E-64 during their exposure to free-fall, K-ras codon 12 base substitution mutation was detected after UV irradiation, although the mutation was not detected after UV irradiation alone. Thus, the increase in E-64-sensitive protease activity may be involved in the suppression of UV mutagenicity in UVAP-1 cells exposed to free-fall.

Antipain↗
PubMed2003

Focused Assessment with Sonography for Trauma in weightlessness: a feasibility study.

BACKGROUND: The Focused Assessment with Sonography for Trauma (FAST) examines for fluid in gravitationally dependent regions. There is no prior experience with this technique in weightlessness, such as on the International Space Station, where sonography is currently the only diagnostic imaging tool. STUDY DESIGN: A ground-based (1 g) porcine model for sonography was developed. We examined both the feasibility and the comparative performance of the FAST examination in parabolic flight. Sonographic detection and fluid behavior were evaluated in four animals during alternating weightlessness (0 g) and hypergravity (1.8 g) periods. During flight, boluses of fluid were incrementally introduced into the peritoneal cavity. Standardized sonographic windows were recorded. Postflight, the video recordings were divided into 169 20-second segments for subsequent interpretation by 12 blinded ultrasonography experts. Reviewers first decided whether a video segment was of sufficient diagnostic quality to analyze (determinate). Determinate segments were then analyzed as containing or not containing fluid. A probit regression model compared the probability of a positive fluid diagnosis to actual fluid levels (0 to 500 mL) under both 0-g and 1.8-g conditions. RESULTS: The in-flight sonographers found real-time scanning and interpretation technically similar to that of terrestrial conditions, as long as restraint was maintained. On blinded review, 80% of the recorded ultrasound segments were considered determinate. The best sensitivity for diagnosis in 0 g was found to be from the subhepatic space, with probability of a positive fluid diagnosis ranging from 9% (no fluid) to 51% (500 mL fluid). CONCLUSIONS: The FAST examination is technically feasible in weightlessness, and merits operational consideration for clinical contingencies in space.

Aerospace Medicine↗
PubMed1997

Activation and proliferation of lymphocytes and other mammalian cells in microgravity.

The experimental findings reviewed in this chapter support the following conclusions: Proliferation. Human T-lymphocytes, associated with monocytes as accessory cells, show dramatic changes in the centrifuge, in the clinostat and in space. In free-floating cells the mitogenic response is depressed by 90% in microgravity, whereas in cells attached to a substratum activation is enhanced by 100% compared to 1-G ground and inflight controls. The duration of phase G1 of the mitotic cycle of HeLa cells is reduced in hypergravity, resulting in an increased proliferation rate. Other systems like Friend cells and WI38 human embryonic lung cells do not show significant changes. Genetic expression and signal transduction. T-lymphocytes and monocytes show important changes in the expression of cytokines like interleukin-1, interleukin-2, interferon-gamma and tumor necrosis factor. The data from space experiments in Spacelab, Space Shuttle mid-deck, and Biokosmos have helped to clarify certain aspects of the mechanism of T-cell activation. Epidermoid A431 cells show changes in the genetic expression of the proto-oncogenes c-fos and c-jun in the clinostat and in sounding rockets. Membrane function, in particular the binding of ligates as first messengers of a signal, is not changed in most of the cell systems in microgravity. Morphology and Mortility. Free cells, lymphocytes in particular, are able to move and form aggregates in microgravity, indicating that cell-cell contacts and cell communications do take place in microgravity. Dramatic morphological and ultrastructural changes are not detected in cells cultured in microgravity. Important experiments with single mammalian cells, including immune cells, were carried out recently in three Spacelab flights, (SL-J, D-2, and IML-2 in 1992, 1993, and 1994, respectively). The results of the D-2 mission have been published in ref. 75; those of the IML-2 mission in ref. 76. Finally, many cell biology experiments in space have suffered in the past from a lack of adequate controls (like 1-G centrifuges) and of proper experimental conditions (like well-controlled temperature). In this respect the availability of Biorack, outfitted with proper incubators with 1-G control centrifuge as well as a glovebox with a microscope, is a great advantage. It is also desirable that cell biology experiments in space are accompanied or even preceded by a program of ground-based investigations in the fast rotating clinostat and in the centrifuge, and that preparatory experiments be done in parabolic flights and sounding rockets, whenever possible. Proper publication of the results of space experiments is another important need. A great number of data have been published in proceedings and reports that are not available to the broad scientific community. To guarantee the credibility and the international recognition of space biology it is important that the results be published in international, peer reviewed journals.

Animals↗
PubMed1998

A nude mouse model for human bone formation in unloaded conditions.

We describe an experimental model for human bone formation in unloaded conditions. Bone formation has been assessed by implanting in vivo human bone marrow stromal cells (BMSC) on porous hydroxyapatite (HA) bioceramics subcutaneously in nude mice. In this system, human bone formation and remodeling occurs and can be studied in unloaded conditions, i.e., with no influence of muscle tension. Using this model system, we have been also studying the effects of dexamethasone (Dex) in combination with fibroblast growth factor-2 (FGF-2) on the osteogenic potential of human BMSC. A colony-forming unit-fibroblastoid (CFU-F) formed in clonal conditions were significantly larger when Dex/FGF-2 was present in the culture medium. The cell proliferation rate was also increased by the combination Dex/FGF-2 at a higher extent than Dex or FGF-2 alone. BMSC expanded with Dex/FGF-2 displayed alkaline phosphatase levels lower (56%) than Dex expanded cells, but significantly higher than FGF-2 expanded cells. Our results suggest that Dex/FGF-2 expanded BMSC are able to form more bone than BMSC expanded in the presence of FGF-2 alone.

Adult↗
PubMed2005

Real-time QCM-D immunoassay through oriented antibody immobilization using cross-linked hydrogel biointerfaces.

This report presents the development of pre-cross-linked and in situ cross-linked polyethyleneimine-carboxymethylcellulose antibody immobilization platforms for real-time QCM-D immunoassay of sepsis-related biomarkers. These platforms differ significantly from recent trends in QCM-based assays, a rapidly expanding field given the affordability and sensitivity of the transduction system, by providing ultrafast biointerface deposition through cross-linking of polysaccharides. Using rhIL-1ra (17 kDa), a known sepsis biomarker, for development, various immunoassay modifications to increase sensitivity were investigated, including the use of Protein A, Protein G, and anti-IgG Fc specific antibody capture ligands for oriented antibody immobilization, higher-frequency QCM-D crystals, and amplification using secondary antibodies. The optimized assay employs Protein A oriented immobilization on pre-cross-linked polymer and secondary antibodies to achieve a detection limit of 25 ng/mL on 5 MHz crystals. Assay repeatability using the optimized chemistry is robust, with no loss in 100 ng/mL antigen detection over 20 cycles of the 10 min sandwich assay. Nonspecific adsorption of human serum albumin, as characterized by ToF-SIMS, is minimal and negligible for the pre-cross-linked and in situ cross-linked compositions, respectively.

Antibodies↗
PubMed2003

Characteristic of changes in the structure and metabolism of the vastus lateralis muscles in monkeys after space flight.

Monkeys subjected to space flight were found to have significant decreases in the sizes of slow and rapid fibers in the vastus lateralis muscle, due not only to weightlessness but also, to some extent, to restriction of movement activity within the capsule. The quantity of total protein in muscle fibers did not decrease. The respiratory peak in the pool of vastus lateralis muscle fibers decreased after space flight, as did the activity of oxidative enzymes (particularly in rapid fibers of the vastus lateralis muscle).

Animals↗
PubMed2003

Effects of weightlessness and movement restriction on the structure and metabolism of the soleus muscle in monkeys after space flight.

After humans and animals have been in conditions of real and modeled weightlessness, the most marked changes are seen in the "slow" tonic muscles, particularly soleus. Studies of the effects of weightlessness and movement restriction on the soleus muscle in monkeys demonstrated significant reductions in the sizes of slow and rapid fibers due mainly to the actions of real weightlessness (rather than movement restriction in the space capsule). Protein loss in soleus muscle fibers in monkeys following space flight was more marked than loss of other components, including water. The level of atrophy of soleus muscle fibers in these conditions was greater than the decrease in the number of capillaries. Succinate dehydrogenase activity in soleus muscle fibers decreased proportionally to the reduction in fiber size.

Animals↗