Growth analysis of mice subjected to high gravity.
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Animals↗
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Application of multistage continuous fermentation for production of fuel alcohol by very-high-gravity fermentation technology.
A fermentation system to test the merging of very-high-gravity (VHG) and multistage continuous culture fermentation (MCCF) technologies was constructed and evaluated for fuel ethanol production. Simulated mashes ranging from 15% to 32% w/v glucose were fermented by Saccharomyces cerevisiae and the dilution rates were adjusted for each glucose concentration to provide an effluent containing less than 0.3% w/v glucose (greater than 99% consumption of glucose). The MCCF can be operated with glucose concentrations up to 32% w/v, which indicates that the system can successfully operate under VHG conditions. With 32% w/v glucose in the medium reservoir, a maximum of 16.73% v/v ethanol was produced in the MCCF. The introduction of VHG fermentation into continuous culture technology allows an improvement in ethanol productivity while producing ethanol continuously. In comparing the viability of yeast by methylene blue and plate count procedures, the results in this work indicate that the methylene blue procedure may overestimate the proportion of dead cells in the population. Ethanol productivity (Yps) increased from the first to the last fermentor in the sequence at all glucose concentrations used. This indicated that ethanol is more effectively produced in later fermentors in the MCCF, and that the notion of a constant Yps is not a valid assumption for use in mathematical modeling of MCCFs.
Biomass↗
Cross spectral density analysis of heart rate and blood pressure modulation during sinusoidal acceleration in the human centrifuge.
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Adult↗
Neurovestibular modulation of circadian and homeostatic regulation: vestibulohypothalamic connection?
Chronic exposure to increased force environments (+G) has pronounced effects on the circadian and homeostatic regulation of body temperature (T(b)), ambulatory activity (Act), heart rate, feeding, and adiposity. By using the Brn 3.1 knockout mouse, which lacks vestibular hair cells, we recently described a major role of the vestibular system in mediating some of these adaptive responses. The present study used the C57BL6JEi-het mouse strain (het), which lacks macular otoconia, to elucidate the contribution of specific vestibular receptors. In this study, eight het and eight WT mice were exposed to 2G for 8 weeks by means of chronic centrifugation. In addition, eight het and eight WT mice were maintained as 1G controls in similar conditions. Upon 2G exposure, the WT exhibited a decrease in T(b) and an attenuated T(b) circadian rhythm. Act means and rhythms also were attenuated. Body mass and food intake were significantly lower than the 1G controls. After 8 weeks, percent body fat was significantly lower in the WT mice (P < 0.0001). In contrast, the het mice did not exhibit a decrease in mean T(b) and only a slight decrease in T(b) circadian amplitude. het Act levels were attenuated similarly to the WT mice. Body mass and food intake were only slightly attenuated in the het mice, and percent body fat, after 8 weeks, was not different in the 2G het group. These results link the vestibular macular receptors with specific alterations in homeostatic and circadian regulation.
Acceleration↗
Physiological parameters of gravitaxis in the flagellate Euglena gracilis obtained during a parabolic flight campaign.
The unicellular freshwater flagellate Euglena gracilis and its close relative Astasia longa show a pronounced negative gravitaxis. Previous experiments revealed that gravitaxis is most likely mediated by an active physiological mechanism in which changes of the internal calcium concentration and the membrane potential play an important role. In a recent parabolic flight experiment on board an aircraft (ESA 29th parabolic flight campaign), changes of graviorientation, membrane potential and the cytosolic calcium concentration upon changes of the acceleration (between 1 x g(n), 1.8 x g(n), microgravity) were monitored by image analysis and photometric methods using Oxonol VI (membrane potential) and Calcium Crimson (cytosolic calcium concentration). The parabolic flight maneuvers performed by the aircraft resulted in transient phases of 1.8 x g(n) (about 20 s), microgravity (about 22 s) followed by 1.8 x g(n) (about 20 s). A transient increase in the intracellular calcium concentration was detected from lower to higher accelerations (1 x g(n) to 1.8 x g(n) or microgravity to 1.8 x g(n)). Oxonol VI-labeled cells showed a signal, which indicates a depolarization during the transition from 1 x g(n) to 1.8 x g(n), a weak repolarization in microgravity followed by a rapid repolarization in the subsequent 1 x g(n) phase. The results show good coincidence with observations of recent terrestrial and space experiments.
Acceleration↗
Laser in situ keratomileusis flap stability during simulated aircraft ejection in a rabbit model.
PURPOSE: To determine the stability of the laser in situ keratomileusis (LASIK) flap in a rabbit model when subjected to vertical acceleration at nine times the force of gravity (+9 Gz) in an aircraft cockpit ejection simulator. METHODS: Thirty-six eyes from 25 New Zealand white rabbits underwent LASIK flap creation without laser photoablation. One month after surgery, the rabbits were sedated and harnessed in a cockpit ejection seat simulator used to train United States Air Force pilots. They then underwent a controlled rapid-sequence ejection at +9 Gz. Subsequently, the rabbits were euthanized and the corneas harvested for microscopic examination. Refractive measurements and corneal examination were made before LASIK flap creation and prior to and after the +9 Gz ejections. Determination of LASIK flap dislocation was based on clinical observation of flap slippage or a significant shift in pre-ejection to postejection cylinder axis. RESULTS: The average preoperative refraction of the rabbit eye was +1.83 D + 3.25 D x 086 degrees. The average change from pre-ejection to postejection was 0.04 D sphere, 0.02 D cylinder, 6.8 axis degrees, and 0.04 D spherical equivalent. The pre-ejection to postejection measurements were not statistically significantly different by a paired test. Laser in situ keratomileusis flap dislocation or ejection-induced corneal folds or striae were not clinically observed. Histologic examination revealed well-healed LASIK flaps but no reactive keratocytes at the central stromal-stromal interface. CONCLUSIONS: Healed LASIK flaps as created in this rabbit model without laser ablation are stable when subjected to a rapid vertical ejection at nine times the force of gravity.
Aerospace Medicine↗
Evolution of bioconvective patterns in variable gravity.
Measurements are reported of the evolution of bioconvective patterns in shallow, dense cultures of microorganisms subjected to varying gravity. Various statistical properties of this random, quasi-two-dimensional structure have been found: Aboav's law is obeyed, the average vertex angles follow predictions for regular polygons, and the area of a pattern varies linearly with its number of sides. As gravity varies between 1 g and 1.8g (g = 9.8 m s-1), these statistical properties continue to hold despite a tripling of the number of polygons and a reduced average polygon dimension by a third. This work compares with experiments on soap foams, Langmuir monolayer foams, metal grains, and simulations.
Animals↗
Circumnutations: from Darwin to space flights.
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Centrifugation↗
Plant cell growth responds to external forces and the response requires intact microtubules.
Microfibril deposition in most plant cells is influenced by cortical microtubules. Thus, cortical microtubules are templates that provide spatial information to the cell wall. How cortical microtubules acquire their spatial information and are positioned is unknown. There are indications that plant cells respond to mechanical stresses by using microtubules as sensing elements. Regenerating protoplasts from tobacco (Nicotiana tabacum) were used to determine whether cells can be induced to expand in a preferential direction in response to an externally applied unidirectional force. Additionally, an anti-microtubule herbicide was used to investigate the role of microtubules in the response to this force. Protoplasts were embedded in agarose, briefly centrifuged at 28 to 34g, and either cultured or immediately prepared for immunolocalization of their microtubules. The microtubules within many centrifuged protoplasts were found to be oriented parallel to the centrifugal force vector. Most protoplasts elongated with a preferential axis that was oriented 60 to 90 degrees to the applied force vector. Protoplasts treated transiently with the reversible microtubule-disrupting agent amiprophos-methyl (applied before and during centrifugation) elongated but without a preferential growth axis. These results indicate that brief biophysical forces may influence the alignment of cortical microtubules and that microtubules themselves act as biophysical responding elements.
Biophysical Phenomena↗
Modeling of dynamic cardiovascular responses during G-transition-induced orthostatic stress in pitch and roll rotations.
Dynamic and fuzzy models for a typical subject's cardiovascular response to the orthostatic stress have been developed based on experimental data. In our original study (Cheung et al., 1999), arterial blood pressure (BP) time-series data were obtained using a man-rated tilt table that applies gigahertz-acceleration transitions from +0.861 Gz [head-up (HU)] to -0.707 G [head-down (HD)] and back to +0.861 Gz (HU) using either pitch or roll rotations (Cheung et al., 1999). G transitions of different duration and onset rates are common in fighter maneuvers. Based on these data, two types of predictive models have been developed in this paper: 1) second-order discrete-time models that predict BP dynamics during pitch and roll rotations and 2) fuzzy logic models that predict important variations in a subject's cardiovascular dynamics induced by HU-to-HD and HD-to-HU transitions. These two types of models assist in providing an operationally important predictive view on the characteristics of BP responses to orthostatic stress induced by pitch and roll rotations of a fighter jet pilot. The new models are being currently utilized in the design of operational recommendations for more G-tolerant operational flight regimes (e.g., split-S tactical maneuver) than the ones currently in use for modern combat aircraft.
Acceleration↗
Influence of the g-force on the circumnutations of sunflower hypocotyls.
Circumnutations of hypocotyls of sunflower (Helianthus annuus L. cv. Californicus) were studied under 1 g and 3 g conditions. Root mean square values of the hypocotyl deviation from the plumbline and period of the movements were determined from calculations of the autocorrelation functions of the movements. The amplitude and the period of the circumnutations increased under 3 g as compared to 1 g. A transition from 3 to 1 g or vice versa also caused changes in period and amplitude of the movements. The results are interpreted as a support for the idea that gravity influences the circumnutation parameters in this sunflower variety. A comparison is made with published results on the dwarf sunflower cv. Teddy Bear where the force influence is very small or negligible. Simulations of a model for circumnutations show movements which are in qualitative agreement with the experimental results, provided adaptation to g-levels is included in the model. Finally, the results are discussed with the recent Spacelab-experiment (SL1) as a background.
Centrifugation↗
Perception and response to gravity in higher fungi--a critical appraisal.
Considering that research on gravitropism in higher fungi has a history of over 100 years, the harvest of established fact is disappointingly meagre. We can be reasonably certain of the following. Hymenomycete 'mushroom' fruit bodies (polypore and agaric) exhibit a number of tropisms of which anemotropism, gravitropism, phototropism and thigmotropism have been clearly demonstrated. At any one time one tropism usually predominates but the inferior tropisms can be demonstrated if the predominating ones can be removed by manipulation of the growth conditions. In ascending order, the hierarchy appears to be: thigmotropism, gravitropism, anemotropism, phototropism. During the course of development of a fruit body different tropisms predominate at different times. The youngest fruit body initials grow perpendicularly away from their substratum. The nature of this tropism is completely unknown but perpendicular growth of fruit body initials has been remarked upon in experiments at a variety of light intensities and in gravitational fields from +/- 0 to 4.5 g. The fruit-body primordium then becomes first positively phototropic but later negative gravitropism predominates. The switch between predominance of the two tropisms has been associated with the onset of sporulation in a number of different studies. The major adjustment of the direction of growth in response to a tropic stimulus is made by the mushroom stem. It is the apex of the stem which makes the most immediate gravitropic response. Gravitropic growth curvatures are limited to the normal growth zones of the stem and seem to depend on re-allocation of available growth resources. If the fruit body is reoriented late in the growth of the stem, it may not be able to respond fully. In these cases gravitropic movements of the cap may still be able to bring the hymenophore back to the vertical. Mechanical forces may influence and contribute to the 'gravitropic' response but this has not been experimentally examined. The hymenophore (gill, tube or tooth) is positively gravitropic and responds independently of the stem. Bracket polypores do not show tropisms but exhibit gravimorphogenetic responses such that gross disturbance leads to renewal of growth to produce and entirely new fruiting structure suitably reoriented to the new spatial position. One experiment performed on an orbiting space station suggests that, in the absence of a light stimulus, gravity may be required for initiation of fruiting in Polyporus brumalis. Otherwise, the indications from both clinostat and space-borne experiments are that the basic form of the mushroom (overall tissue arrangement of stem, cap, gills, hymenium, veil) in agaric and polypore alike is established independently of the gravity vector. Abnormal stem growth has been observed in clinostat cultures of Panus (= Lentinus) tigrinus and Polyporus brumalis, but the morphogenetic event which seems most dependent on gravity is sporulation (in the broadest sense). Cultures of P. brumalis on orbiting space craft fail to produce the poroid hymenophore and in clinostat experiments on the ground even karyogamy was rare in similar cultures. Coprinus cinereus grown on the clinostat was able to produce apparently normal fruit body primordia which failed to produce spores and then aborted, forming a new flush of primordia on the old. Taken together with the clear association between observation of gravitropism and the onset of sporulation, the implication is that commitment to the meiosis-sporulation pathway both requires the gravity vector and couples it in some way to fruit-body growth. There is no convincing evidence for a graviperception mechanism in fungi. There is no evidence for any organised means of communicating the gravitropic stimulus once it has been perceived. Reports of three different experimental studies reveal the authors' conviction that the apparently coordinated expression of gravitropic response is in truth a common, but independent, response by the individual component hyphae of the structure concerned. There is some evidence that in the negatively gravitropic Phycomyces sporangiophore the vacuole floats in the protoplasm. If this is generally true it could affect protoplasmic volumes above and below the vacuole such that a greater proportion of the cell's potential for wall growth was adjacent to the lower wall. This is not only an attractive way of accounting for asymmetric wall growth, but since the relative density of the vacuole can presumably be controlled by regulation of water influx and efflux, it is also an attractive means of accounting for the control of gravitropic responses. Phycomyces also exhibits a response to the mechanical consequences of reorientation which is additional to (and different from) the longer term gravitropic response. [TRUNCATED]
Basidiomycota↗
Improvement of ethanol production in very high gravity fermentation by horse gram (Dolichos biflorus) flour supplementation.
AIMS: To determine the effect of osmotic stress on yeast and to investigate the protective role of horse gram flour during very high gravity (VHG) ethanol fermentation. METHODS AND RESULTS: Saccharomyces cerevisiae was inoculated into high sugar (30-40%, w/v) containing medium with and without supplementation of horse gram flour. The fermentation experiments were carried out in batch mode. The effect of 4 or 6% of horse gram flour to the medium on the metabolic behaviour and viability of yeast was studied. Significant increase in ethanol yield up to 50% and dramatic decrease in glycerol production up to 100% was observed in the presence of horse gram flour. The fermentation rate was increased from 3 to 5 days with increased viable cell count. The physical and chemical factors of horse gram flour may aid in reducing the osmotic stress of high gravity fermentation of ethanol as well as enhancing ethanol yield. CONCLUSIONS: It was found that horse gram flour not only reduced fermentation time but also enhanced ethanol production by better utilization of sugar. SIGNIFICANCE AND IMPACT OF THE STUDY: Production of high ethanol concentration by using VHG sugar fermentation eliminates the expensive steps in the conventional process and saves time.
Dolichos↗
Orientation of Paramecium under the conditions of weightlessness.
A cell culture of Paramecium with a precise negative gravitaxis was exposed to 4 x l0(-6) g during a parabolic flight of a sounding rocket for 6 min. Computer image analysis revealed that without gravity stimulus the individual swimming paths remained straight. In addition, three reactions could be distinguished. For about 30 s, paramecia maintained the swimming direction they had before onset of low gravity. During the next 20 s, an approximate reversal of the swimming direction occurred. This period was followed by the expected random swimming pattern. Similar behavior was observed under the condition of simulated weightlessness on a fast-rotating clinostat. Control experiments on the ground under hyper-gravity on a low-speed centrifuge microscope and on a vibration test facility proved that the observed effects were caused exclusively by the reduction of gravity.
Acceleration↗
Influence of gravity on cardiac performance.
Results obtained by the investigators in ground-based experiments and in two parabolic flight series of tests aboard the NASA KC-135 aircraft with a hydraulic simulator of the human systemic circulation have confirmed that a simple lack of hydrostatic pressure within an artificial ventricle causes a decrease in stroke volume of 20%-50%. A corresponding drop in stroke volume (SV) and cardiac output (CO) was observed over a range of atrial pressures (AP), representing a rightward shift of the classic CO versus AP cardiac function curve. These results are in agreement with echocardiographic experiments performed on space shuttle flights, where an average decrease in SV of 15% was measured following a three-day period of adaptation to weightlessness. The similarity of behavior of the hydraulic model to the human system suggests that the simple physical effects of the lack of hydrostatic pressure may be an important mechanism for the observed changes in cardiac performance in astronauts during the weightlessness of space flight.
Biomedical Engineering↗
The gravity field of Mars: results from Mars Global Surveyor.
Observations of the gravity field of Mars reveal a planet that has responded differently in its northern and southern hemispheres to major impacts and volcanic processes. The rough, elevated southern hemisphere has a relatively featureless gravitational signature indicating a state of near-isostatic compensation, whereas the smooth, low northern plains display a wider range of gravitational anomalies that indicates a thinner but stronger surface layer than in the south. The northern hemisphere shows evidence for buried impact basins, although none large enough to explain the hemispheric elevation difference. The gravitational potential signature of Tharsis is approximately axisymmetric and contains the Tharsis Montes but not the Olympus Mons or Alba Patera volcanoes. The gravity signature of Valles Marineris extends into Chryse and provides an estimate of material removed by early fluvial activity.
Atmosphere↗
Isolation and characterization of brewer's yeast variants with improved fermentation performance under high-gravity conditions.
To save energy, space, and time, today's breweries make use of high-gravity brewing in which concentrated medium (wort) is fermented, resulting in a product with higher ethanol content. After fermentation, the product is diluted to obtain beer with the desired alcohol content. While economically desirable, the use of wort with an even higher sugar concentration is limited by the inability of brewer's yeast (Saccharomyces pastorianus) to efficiently ferment such concentrated medium. Here, we describe a successful strategy to obtain yeast variants with significantly improved fermentation capacity under high-gravity conditions. We isolated better-performing variants of the industrial lager strain CMBS33 by subjecting a pool of UV-induced variants to consecutive rounds of fermentation in very-high-gravity wort (>22 degrees Plato). Two variants (GT336 and GT344) showing faster fermentation rates and/or more-complete attenuation as well as improved viability under high ethanol conditions were identified. The variants displayed the same advantages in a pilot-scale stirred fermenter under high-gravity conditions at 11 degrees C. Microarray analysis identified several genes whose altered expression may be responsible for the superior performance of the variants. The role of some of these candidate genes was confirmed by genetic transformation. Our study shows that proper selection conditions allow the isolation of variants of commercial brewer's yeast with superior fermentation characteristics. Moreover, it is the first study to identify genes that affect fermentation performance under high-gravity conditions. The results are of interest to the beer and bioethanol industries, where the use of more-concentrated medium is economically advantageous.
Beer↗
Analysis of muscular development of mice at high gravity.
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Animals↗