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Needs of physiological and psychological research using artificial gravity.

In the next century, mankind will expand its activity to the moon and Mars. At that time, humans will be exposed to a low and micro-gravity environment in long term which causes physiological and psychological problems. The authors propose an artificial gravity space station for a research laboratory on human physiology and psychology at various gravity levels. The baseline specifications and the configuration of the space station are shown. Reviewing the history of manned space flight, the necessity of the research on an artificial gravity space station is discussed, including themes of research to be conducted on the station and the application of its results. Technical issues for realization of the space station such as environmental factors, system function and assembly scenario are also discussed.

Aerospace Medicine↗

Cell biology of plant gravity sensing.

The debate about whether gravity sensing relies upon statoliths (amyloplasts that sediment) has intensified with recent findings of gravitropism in starchless mutants and of claims of hydrostatic gravity sensing. Starch and significant plastid sedimentation are not necessary for reduced sensing in mutant roots, but plastids might function here if there were a specialized receptor for plastid mass e.g. in the ER. Alternatively, components in addition to amyloplasts might provide mass for sensing. The nucleus is dense and its position is regulated, but no direct data exist for its role in sensing. If the weight of the protoplast functioned in sensing, why would there be specific cytological specializations favoring sedimentation rather than cell mass? Gravity has multiple effects on plants in addition to gravitropism. There may be more than one mechanism of gravity sensing.

Cell Nucleus↗

The effect of gravity on plant germination.

An axis clinostat was constructed to create micro and negative gravity also a rotated flat disk was constructed with different rotation rates to give increased gravity, by centrifugal force up to 48 g. Rice seeds were grown on agar in tubes at the constant air temperature of 20 degrees C under an average light condition of 110 micromol/m2/sec(PPF). Humidity was not controlled but was maintained above 90%. Since the tube containers were not large enough for long cultivation, shoot and root growth were observed every 12 hours until the sixth day from seeding. The lengths of shoots and roots for each individual plant were measured on the last day. The stem lengths were increased by microgravity but the root lengths were not. Under the negative gravity, negative orthogeotropism and under microgravity, diageotropism was observed. No significant effect of increased gravity was observed on shoot and root growth.

Centrifugation↗

The role of gravity in the phylogeny of structure and function in animal sensors of spatial orientation, and their predicted action in weightlessness.

The evolution of the structural, functional and cytochemical organization of the gravity receptor which determines a body position in the gravitational field of the earth by means of muscular regulation was traced both invertebrates and vertebrates, using electron microscopic and histochemical methods. In the course of evolution of vertebrates, the specialized gravity receptor-statocyst which, as a rule, consists of primary sensory cells and supplies otoliths, is formed. In vertebrates, there exists a vestibular apparatus made up of secondary sensory cells and also having otoliths. The receptor cells, both of statocysts and the vestibular apparatus, are supplied with special antennas (kinocilia and stereocilia). Deviation of the antennas stimulated by displacement of the otoliths resulting from locomotor activity of animals leads to excitation of the receptor cells. When exposed to a modified gravitational field (linear accelerations of 10 g, for 3 min), the receptor cells of the vestibular apparatus, in all classes of vertebrates, show progressive changes in RNA content and protein synthesis (increase followed by decrease) which return to normal only after 12 days. Thus, immediate transfer of animals and man from acceleration to weightlessness appears to be a reason for movement disease. The above consideration showed the need for an experiment in which an animal (with its vestibular apparatus) which had not undergone previous accelerations, would be exposed to weightlessness. Frog embryos, Rana temporaria, at the stage preceding the organogenesis, when the vestibular apparatus and other organs were lacking, were chosen as a suitable subject. Frog embryos at the stage of an early gastrula were placed in a special container Emkon aboard the Soyuz 10 spacecraft. After short accelerations, they were exposed to weightlessness for 44 hours. The embryos were allowed to continue to develop to the stage of early tail bud. The experimental embryos showed normally developed acoustic vesicles and vestibular ganglia. Clear differentiation of the receptor cells with antennas (kinocilia and stereocilia) was found in the acoustic vesicles. Thus, in weightlessness, vestibular apparatus develops just as well as in the gravitational field of the earth. However, only a much longer stay in weightlessness conditions will indicate whether there are any changes in the structural, cytochemical and functional organization of vestibular apparatus. The similarity in the structural, functional and cytochemical organization of the gravity receptor in vertebrates and invertebrates appears to allow the prediction of the behaviour of the gravity receptor as a whole, and of its receptor elements, both in normal and changed gravitational fields. The first attempts were carried out only on the vestibular apparatus of vertebrates.

Acetylcholinesterase↗

Gravity receptor function in mice with graded otoconial deficiencies.

The purpose of the present study was to examine gravity receptor function in mutant mouse strains with variable deficits in otoconia: lethal milk (lm), pallid (pa), tilted (tlt), mocha (mh), and muted (mu). Control animals were either age-matched heterozygotes or C57BL/6J (abbr. B6) mice. Gravity receptor function was measured using linear vestibular evoked potentials (VsEPs). Cage and swimming behaviors were also documented. Temporal bones were cleared to assess the overall otoconial deficit and to correlate structure and function for lm mice. Results confirmed the absence of VsEPs for mice that lacked otoconia completely. VsEP thresholds and amplitudes varied in mouse strains with variable loss of otoconia. Some heterozygotes also showed elevated VsEP thresholds in comparison to B6 mice. In lm mice, which have absent otoconia in the utricle and a variable loss of otoconia in the saccule, VsEPs were present and average P1/N1 amplitudes were highly correlated with the average loss of saccular otoconia (R = 0.77,p < 0.001). Cage and swimming behavior were not adversely affected in those animals with recordable VsEPs. Most, but not all, mice with absent VsEPs were unable to swim. Some animals were able to swim despite having no measurable gravity receptor response. The latter finding underscores the remarkable adaptive potential exhibited by neurobehavioral systems following profound sensory loss. It also shows that behavior alone may be an unreliable indicator of the extent of gravity receptor deficits.

Animals↗

Effect of creatinine and specific gravity normalization techniques on xenobiotic biomarkers in smokers' spot and 24-h urines.

Renal excretion mechanisms are xenobiotic-specific; therefore, accurate exposure assessment requires an understanding of relationships of xenobiotic biomarker concentration and excretion rate to urine flow, specific gravity and creatinine concentration. Twenty-four-hour urine collection for xenobiotic exposure assessment is considered the "gold standard" procedure. Random spot-urine collection is convenient and minimizes subject compliance concerns but requires that normalization techniques be employed to account for diuresis and diurnal variation in xenobiotic biomarker excretion. This paper examines and makes recommendations concerning normalization techniques and conditions under which spot-urine results most accurately reflect 24-h urine results. Specific gravity, creatinine, and xenobiotic biomarkers were determined in smokers' spot and 24-h urines. Normalization techniques were applied, variance-component analyses were performed to estimate variability, spot urines were pooled mathematically to simulate 24-h urines and analyses of variance were performed to evaluate spot urines' ability to reflect 24-h urine concentrations. For each xenobiotic biomarker concentration, log-linear relationships were observed with urine flow, specific gravity, and creatinine. For most xenobiotic biomarker excretion rates, log-linear relationships were observed with urine flow; creatinine, however, was unaffected by urine flow. The conventional creatinine ratio-normalization technique demonstrated greater variability (within-day, between-day and between-subject) than other normalization techniques. Comparisons of simulated 24-h urines to spot urines suggest that spot-urine collection be performed only between 2 p.m. and 2 a.m. and that the modified specific-gravity-adjusted-creatinine ratio-normalization technique and the creatinine-regression normalization technique yield the best agreement between spot- and simulated 24-h urine results.

Biomarkers↗

Is the refractometer specific gravity a reliable index for pediatric fluid management?

Serial measurement of urinary output has been one of the mainstays to gauge adequacy of hydration of the newborn surgical patient. Urine output, however, is dependent not only on circulating blood volume but also on osmolar load and the ability of the kidney to excrete solute and water. Therefore, a second measurement is necessary for more accurate monitoring urine concentration. The purpose of this study is to determine the capacity of the kidneys of the newborn surgical patient to dilute and concentrate, and to excrete solute and water; and the accuracy of the refractometer to determine the tonicity of urine. To determine renal capacity, 24 newborn patients were studied during the first 48 hours following major surgical procedures. Analysis of 823 urine samples demonstrated a range of specific gravity between 1.002 and 1.040, and of osmolality between 67 and 582 mosm/kg. The lowest urine output was 0.1 mL/kg/hr and the highest 14.9 mL/kg/hr (mean 3.54 +/- 1.97 mL/kg/hr). The osmolar load excreted by the kidney ranged from 3.7 to 33.4 mosm/kg/24 hr (15.3 +/- 6 mosm/kg/24 hr). To determine the accuracy of the refractometer, 2,007 urine samples from 900 newborn infants were studied. The correlation coefficient for refractometer derived specific gravity and osmolality ranged from 0.7347 to 0.9061. Refractometer specific gravity measurements of 1.008 or below predict hyposmolality as defined as an osmolality below 270 mosm/kg with an accuracy of 70.6% to 95.4%. Accuracy is significantly less in the TPN and the glucose groups. A specific gravity of 1.012 to 1.020 is accurate in predicting hyperosmolality as defined as a urine osmolality above 290 mosm/kg.(ABSTRACT TRUNCATED AT 250 WORDS)

Diuresis↗

A genetic effect of altered gravity: mutations induced by simulated hypogravity and hypergravity in microsatellite sequences of human tumor cells.

To determine the possible genetic effects of gravity alterations, we analyzed mutation induction in microsatellite sequences of human tumor cells treated with simulated hypogravity provided by a clinostat or hypergravity by a centrifuge. Microsatellite mutations were detected as changes in the size of polymerase chain reaction (PCR)-amplified allelic markers. The frequencies of mutant clones in cultures treated with simulated hypogravity for 24 or 48 h were almost the same as those of controls, but after 72 h of treatment, the mutant frequencies had increased significantly in all three microsatellite loci examined. Significantly higher mutant frequencies were similarly detected in cultures treated for 72 h with a hypergravity condition as low as 18xg, but not detected in 24 or 48 h treated cultures. These findings clearly show that gravity alterations that last for 3 days can induce microsatellite mutations in human cells. A genetic effect of gravity change, therefore, is established for the first time. Moreover, high frequencies of microsatellite mutations were induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) which activates protein kinase C-mediated signal transduction pathways and causes genetic instability. These findings suggest that gravity change induces microsatellite mutations by modulating the pattern of gene expression involved in signal transduction pathways.

DNA Primers↗

On the influence of altered gravity on the growth of fish inner ear otoliths.

Inner ear stones (otoliths) of developing cichlid fish (Oreochromis mossambicus) were marked with the calcium tracer alizarin-complexone (AC) at 1g-earth gravity before and after a longterm (20 days) stay of the animals at moderate hypergravity conditions (3g; centrifuge). AC deposition at the otoliths resulted in two fluorescence bands, which enclosed the area grown during exposure to altered gravity. This area was measured with regard to size and asymmetry (size difference between the left and the right stones). Both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure as compared to parallely raised 1 g-control specimens. The asymmetry concerning the lapilli was pronouncedly decreased in comparison to the 1g-controls. These findings suggest, that the growth and the development of bilateral asymmetry of otoliths is guided by the environmental gravity vector. Some of the hyper-g animals revealed a kinetotic behaviour at the transfer from hyper-g to normal 1g-earth gravity conditions, which was qualitatively similar to the behaviour observed in previous experiments at the transfer from 1 g to microgravity in the course of parabolic aircraft flights. The lapillar asymmetry of kinetotic samples was found to be significantly higher than that of normally behaving experimental specimens. This result supports an earlier theoretical concept, according to which human static space sickness might be based on asymmetric utricular otoliths

Animals↗

Neuroplastic reactivity of fish induced by altered gravity conditions: a review of recent results.

A review is being presented concerning behavioural, biochemical, histochemical and electronmicroscopical data on the influence of altered gravitational forces on the swimming performance and on the neuronal differentiation of the brain of cichlid fish larvae and adult swordtail fish that had been exposed to hyper-gravity (3g in laboratory centrifuges), hypo-gravity (>10(-2) g in a fast-rotating clinostat) and to near weightlessness (10(-4) g aboard the Spacelab D-2 mission). After long-term alterations of gravity (and parallel light deprivation), initial disturbances in the swimming behaviour followed by a stepwise regain of normal swimming modes are induced. Parallel, neuroplastic reactivities on different levels of investigation were found, such as adaptive alterations of activities of various enzymes in whole brain as well as in specific neuronal integration centers and an intraneuronal reactivity on ultrastructural level in individual brain parts and in the sensory epithelia of the inner ear. Taken together, these data reveal distinct adaptive neuroplastic reactions of fish to altered gravity conditions.

Adaptation, Physiological↗

Evaluation of gravity-dependent membrane potential shift in Paramecium.

It is still debated whether or not gravity can stimulate unicellular organisms. This question may be settled by revealing changes in the membrane potential in a manner depending on the gravitational forces imposed on the cell. We estimated the gravity-dependent membrane potential shift to be about 1 mV G-1 for Paramecium showing gravikinesis at 1-5 G, on the basis of measurements of gravity-induced changes in active propulsion and those of propulsive velocity in solutions, in which the membrane potential has been measured electrophysiologically. The shift in membrane potential to this extent may occur from mechanoreceptive changes in K+ or Ca2+ conductance by about 1% and might be at the limit of electrophysiological measurement using membrane potential-sensitive dyes. Our measurements of propulsive velocity vs membrane potential also suggested that the reported propulsive force of Paramecium measured in a solution of graded densities with the aid of a video centrifuge microscope at 350 G was 11 times as large as that for -29 mV, i.e., the resting membrane potential at [K+]o = 1 mM and [Ca2+]o = 1 mM, and, by extrapolation, that Paramecium was hyperpolarized to -60 mV by gravity stimulation of 100-G equivalent, the value corrected by considering the reduction of density difference between the interior and exterior of the cell in the graded density solution. The estimated shift of the membrane potential from -29 mV to -60 mV by 100-G equivalent stimulation, i.e., 0.3 mV G -1, could reach the magnitude entirely feasible to be measured more directly.

Animals↗

The role of gravity in the evolutionary emergence of multicellular complexity: microgravity effects on arthropod development and aging.

While experiments carried out in Space with isolated cells have shown that eucaryotic cells are able to sense and respond to the absence of gravity by modifying their reactions, experiments in which more complex processes have been investigated, such as Biological Systems undergoing development under Microgravity, have been surprisingly unaffected by the space environment. This can be considered a curious result since all organisms are evolutionarily adapted to the current level of the gravity force in our planet and should eventually change if this parameter will vary in a permanent manner. In fact, the small effects of the modifications in gravity on development in short term experiments may be equivalent to the difficulties in detecting the involvement of other basic physical processes such as diffusion-controled auto-organizative reactions in currently developing biological systems. An apparent exception to this lack of effect is experiments where brine shrimp dormant gastrulae directly exposed to the space environment accumulate developmental defects as a consequence of cosmic irradiation. In this article we discuss the idea that at a certain stage during the evolutionary emergence of multicellular organisms the cues laid by generic forces such as gravity were involved in the evolutionary organization of these primitive organisms. As evolution proceed, these early mechanisms may have been obscured and/or made redundant by the appearance of new internal, environment-independent biological regulatory mechanisms. On the other hand, behavioral responses that may be important, for example, in setting the life-spans of organisms may still be more readily susceptible to manipulation by external cues as experiments carried out by our group in Space and on the ground with Drosophila melanogaster indicate. Grant Numbers: ESP-1775.

Animals↗

Humans use internal models to estimate gravity and linear acceleration.

Because sensory systems often provide ambiguous information, neural processes must exist to resolve these ambiguities. It is likely that similar neural processes are used by different sensory systems. For example, many tasks require neural processing to distinguish linear acceleration from gravity, but Einstein's equivalence principle states that all linear accelerometers must measure both linear acceleration and gravity. Here we investigate whether the brain uses internal models, defined as neural systems that mimic physical principles, to help estimate linear acceleration and gravity. Internal models may be used in motor contro, sensorimotor integration and sensory processing, but direct experimental evidence for such models is limited. To determine how humans process ambiguous gravity and linear acceleration cues, subjects were tilted after being rotated at a constant velocity about an Earth-vertical axis. We show that the eye movements evoked by this post-rotational tilt include a response component that compensates for the estimated linear acceleration even when no actual linear acceleration occurs. These measured responses are consistent with our internal model predictions that the nervous system can develop a non-zero estimate of linear acceleration even when no true linear acceleration is present.

Acceleration↗

Use of urine specific gravity to improve screening for albuminuria.

The albumin to creatinine ratio (ACR) can be used to measure urine albumin excretion rates, but is inconvenient and expensive. More rapid and less expensive screening methods estimate only albumin concentration and are subject to errors caused by variation in urine volume. We examined whether urine specific gravity could be used in place of urine creatinine to correct albumin concentration for differences in urine volume in 50 patients. Urine specific gravity accurately estimated urine creatinine concentration (r = 0.79, P < 0.001). The albumin estimated-creatinine ratio (ACestR) in random spot urine sample correlated with urine albumin excretion measured in a 24-hour urine collection (r = 0.98, P < 0.001), as did the ACR (r = 0.95, P < 0.001). For determining microalbuminuria, the sensitivity (0.88) and specificity (0.93) of the ACestR were similar to those of ACR (0.89 and 0.93, respectively). Unfortunately, the sensitivity (0.63) of the Micral-Test was relatively poor, and was only slightly improved by correcting for urine specific gravity (0.69) in this small sample of patients. Nevertheless, these results suggest that as rapid methods for measuring urine albumin concentration improve, combining them with urine specific gravity might produce a less expensive and more convenient alternative to the ACR.

Adult↗

Urine specific gravity and water hardness in relation to urolithiasis in persons with spinal cord injury.

STUDY DESIGN: A matched case-control study. OBJECTIVES: To clarify the influence of urine specific gravity and drinking water quality on the formation of urinary stones in persons with spinal cord injury (SCI). SETTING: A rehabilitation center within a university hospital. METHODS: Between 1992 and 1998, 63 stone cases (31 kidney, 27 bladder, and five both) and 289 age-duration-matched controls were recruited from a cohort of SCI patients enrolled in an on-going longitudinal study. Data on urine specific gravity and other characteristics of study participants were retrieved from the database and medical charts. Community water supply information was provided by the Alabama Department of Environmental Management. Multivariable conditional logistic regression analysis was performed to evaluate the association with stone formation. RESULTS: SCI individuals who had urinary stones were more likely than control subjects to use indwelling catheters and have decreased renal function. The occurrence of stones was not significantly related to gender, race, severity of injury, urinary tract infection, nor urine pH. After controlling for the potential confounding from other factors, a continuously increasing stone occurrence with increasing specific gravity was observed (P=0.05); this association was stronger for kidney (odds Ratio [OR]=1.8 per 0.010 g/cm(3)) versus bladder stones (OR=1.2) and for recurrent (OR=2.0) versus first stones (OR=1.5). Increased water hardness was not significantly associated with a decreased stone occurrence. CONCLUSIONS: Study results suggest that maintaining urine specific gravity below a certain level might reduce the occurrence of urinary stones. This could be easily achieved by using a dipstick for self-feedback along with appropriate fluid intake. For persons with SCI who are at an increased risk of a devastating stone disease, this prophylactic approach could be very cost-effective; however, this requires further confirmation.

Adult↗

A SNARE complex containing SGR3/AtVAM3 and ZIG/VTI11 in gravity-sensing cells is important for Arabidopsis shoot gravitropism.

Plants can sense the direction of gravity and change the growth orientation of their organs. The molecular mechanisms of gravity sensing and signal transduction during gravitropism are not well known. We have isolated several shoot gravitropism (sgr) mutants of Arabidopsis. The sgr3-1 mutant exhibits a reduced gravitropic response in the inflorescence stems. In the inflorescence stems of Arabidopsis, gravity is sensed in endodermal cells that contain sedimentable amyloplasts. In sgr3-1, some amyloplasts in the endodermis failed to sediment in the direction of gravity. SGR3 encodes a syntaxin, AtVAM3, which had previously been cloned as a homologue of yeast Vam3p. AtVAM3 is localized to the prevacuolar compartment and vacuole and is suggested to function in vesicle transport to the vacuole. We have also cloned another soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE), ZIG/AtVTI11, a mutation that causes abnormal gravitropism. This mutant displayed an abnormal distribution of amyloplasts in the endodermal cells similar to that in sgr3-1. Endodermis-specific expression of SGR3 and ZIG by using the SCR promoter could complement the abnormal shoot gravitropism of each mutant. Protein-protein interaction between AtVAM3 and AtVTI11 in the endodermal cells was detected immunologically. The sgr3-1 mutation appeared to reduce the affinity of AtVAM3 for AtVTI11 or SYP5. These results suggest that vesicle transport to the prevacuolar compartment/vacuole in the endodermal cells, mediated by a specific SNARE complex containing AtVAM3 and AtVTI11, plays an important role in shoot gravitropism.

Amino Acid Sequence↗

Modelling the relationships of egg weight, specific gravity, shell calcium and shell thickness.

1. The relationships between egg weight, egg specific gravity, shell weight, shell calcium and shell thickness of 800 eggs from 8 treatments were expressed using mathematical models. 2. The equations describing the relationships were on the basis of any two independent variables predicting the remainder. 3. Of 10 possible models, 4 had high co-efficients of determination (R2 greater than 0.80) for each predicted dependent variable. 4. The two independent variables in each of these 4 models were, in turn, egg weight and specific gravity, egg weight and shell weight, egg weight and shell thickness, and specific gravity and shell weight. 5. The best model was that having egg weight and specific gravity as independent variables, with R2 values of 0.94, 0.88, and 0.85 for predicted shell weight, shell calcium, and shell thickness, respectively. Moreover, egg characteristics can be measured non-destructively by this model, whereas the other three require destruction of the egg.

Age Factors↗

Rapid microalbuminuria screening in type 2 diabetes mellitus: simplified approach with Micral test strips and specific gravity.

BACKGROUND: Microalbuminuria is known to be a harbinger of serious complications in type 2 diabetes mellitus. Since medical intervention at the onset of microalbuminuria can be critical in reducing these adverse outcomes, it is widely agreed that type 2 diabetic patients should be screened for microalbuminuria. The purpose of the present study is to evaluate Micral test strips in conjunction with a urine specific gravity determination as a rapid and accurate method for detecting microalbuminuria in type 2 diabetic patients. METHODS: In this prospective study, a total of 444 urine samples of type 2 diabetic patients were obtained from the ABCD study cohort for analysis. Urinary albumin concentrations were determined using Micral test strips and compared to results measuring albumin by the immunoturbidimetry method of timed collections. Urine specific gravity was measured by a standard urine dipstick. RESULTS: The performance characteristics of the Micral test strips for detecting microalbuminuria (30-300 mg albumin/24 h) were adequate but not optimal: sensitivity 88%, specificity 80%, positive predictive value 69%, negative predictive value 92%. A concomitant specific gravity determination was useful in indexing the magnitude of false negative and false positive readings by the Micral test strips. CONCLUSIONS: While the use of Micral test strips provides a rapid approach to detecting microalbuminuria in type 2 diabetic patients, this method has limitations. The simultaneous measurement of specific gravity is helpful in addressing some of the shortcomings of this screening test.

Albuminuria↗