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Behavioural and biochemical investigations of the influence of altered gravity on the CNS of aquatic vertebrates during ontogeny.

Quantitative data are presented on the influences of hyper-gravity (3 +/- 1g) and of simulated weightlessness (approximately 0g) during early ontogeny of cichlid fish (Oreochromis mossambicus) and clawed toad (Xenopus laevis, Daudin) demonstrating changes in the swimming behaviour and the brain energy and plasma membrane metabolism. After return to 1g conditions, hyper-g reared fish and toads express the well known "loop-swimming" behaviour. By means of a computer based video analyzing system different types of swimming movements and velocities were quantitatively determined. Analyses of the brain energy and plasma-membrane metabolism of hyper-g fish larvae demonstrated an increase in energy availability (glucose 6Pi dehydrogenase, G-6P-DH), a decrease of cellular energy transformation (creatine kinase activity, CK) but no changes in energy consumptive processes (e.g. ATPases) and cytochrome oxidase activity (Cyt.-Ox). In contrast hypo-g fish larvae showed a slight increase in brain CK activity. In addition, unlike 1g controls, hyper-g fish larvae showed pronounced variations in the composition (=polarity) of sialoglycosphingolipids (=gangliosides), typical constituents of the nerve cell membranes, and a slight increase in the activity of sialidase, the enzyme responsible for ganglioside degradation.

Animals↗

Artificial gravity as a countermeasure of physiological deconditioning in space.

The displacement of humans from our planet to space and back to our planet or to another planet necessarily causes the exposure to transitional gravitational forces which create a need for countermeasures of physiological deconditioning. One of the alternative approaches to counteracting physiological deconditioning is the simulation of a gravitational field. An apparatus named the artificial gravity simulator (AGS) has been designed and constructed. The AGS is a short radius rotating platform capable of producing a +Gz 100% gradient of a variable magnitude up to 3-g. It supports four beds for testing four subjects at a time. The AGS is suitable for short and long duration experiments on various physiological systems. At present it is being used to study the cardiovascular response to various g-levels and exposure times.

Cardiovascular Deconditioning↗

Gravity sensing in tip-growing cells.

In addition to the statocytes of roots and shoots, a number of tip-growing cells also sense gravity, which influences the cells' growth and development. Since these tip-growing cells are highly suitable for observations in vivo, the movement and sedimentation of their statoliths can be studied in detail. Experimental manipulation by centrifugation, drug application, optical tweezers or microgravity can be monitored by light microscopy. The statoliths are localized in distinct cytoplasmic areas by interactions with actin filaments or microtubules, and their sedimentation seems to be narrowly confined. Since gravisensing and the graviresponse take place within the same cell, the gravitropic signal transduction chain is not complicated by signal transmission between sensing and responding cells. Studies on tip-growing cells have now enabled the formulation of models explaining positive and negative gravitropism.

Actins↗

Formulas for fetal weight estimation by ultrasound measurements based on neonatal specific gravities and volumes.

To improve the accuracy of fetal weight estimation by ultrasound measurement, especially in low birth weight infants, including those with intrauterine growth retardation, two new formulas were developed from a new point of view. The formulas were established by (1) actual measurements of the volumes of the neonatal head and trunk with limbs, (2) analysis of the relationship between the sizes of the parts of the fetal body measured by ultrasound examination before birth and the volumes of the parts of the neonatal body, and (3) calculation of the specific gravities of the parts of the neonatal body. The prospective study showed a high correlation between estimated weights and actual birth weights with a small range of error in infants weighing between 450 and 4800 gm.

Birth Weight↗

Urinary protein and albumin excretion corrected by creatinine and specific gravity.

Timed urine collections are difficult to use in clinical practice owing to inaccurate collections making calculations of the 24-h albumin or protein excretion questionable. One of our goals was to assess the 'correction' of urinary albumin and (or) protein excretion by dividing these by either the creatinine concentration or the term, (specific gravity-1)x100(1). The 24-h creatinine excretion can be estimated based on the patients' gender, age and weight. We studied the influence of physiological extremes of hydration and exercise, and protein and creatinine excretion in patients with or suspected kidney disorders. Specimens were collected from healthy volunteers every 4 h during one 24-h period. We assayed the collections individually to give us an assessment of the variability of the analytes with time, and then reassayed them after combining them to give a 24-h urine. For all volunteers, the mean intra-individual CVs based on the 4-h collections expressed in mg/24 h were 80.0% for albumin and 96.5% for total protein (P0.2). The CVs were reduced by dividing the albumin or protein concentration by the creatinine concentration or by the term, (SG-1)x100. This gave a CV for mg albumin/g creatinine of 52% (P<0.1 vs. albumin mg/g creatinine); mg protein/g creatinine of 39% (P<0.05 vs. mg protein/g creatinine); mg albumin/[(SG-1)x100] of 49% (P<0.1 vs. albumin)/[(SG-1)x100]; and mg protein/[(SG-1)x100] of 37% (P<0. 05 vs. mg protein)/[(SG-1)x100]. For the 68 subjects in the study, the strongest correlation was between the creatinine concentrations and the 24-h urine volume: r=0.786, P<0.001. The correlation of (SG-1)x100 vs. the 24-h urine volume was: r=0.606, P<0.001; for (SG-1)x100 and the creatinine concentration, the correlation was: r=0.666, P<0.001. Compared to the volunteers, the albumin and protein excretion in mg/24 h were more variable in the patients. The same was true if the albumin or protein concentrations were divided by the creatinine concentration or by (SG-1)x100. Protein and albumin concentrations were lower in dilute urines. Dividing the albumin or protein concentrations by the creatinine concentration reduced the number of false negative protein and albumin results. Dividing the albumin or protein values in mg/24 h by (SG-1)x100 eliminated fewer false negatives. Albumin concentrations increased significantly after vigorous exercise. The increase was almost eliminated when the albumin result was divided by the creatinine concentration suggesting that a decreased urine flow and not increased glomerular permeability causes an increase of post-exercise albuminuria. The same was true for proteinuria. A dipstick test plus an optical strip reader that can measure urine protein, albumin, and creatinine and calculate the appropriate ratios provides a better screening test for albuminuria or proteinuria than one measuring only albumin or protein.

Adult↗

Assessment of specific gravity determined by Ames N-Multistix-SG.

Specific gravity (SG) was determined in 204 urines by N-Multistix-SG solid phase test strip. Seventy-seven of the urines were from patients with known renal disease or were positive for protein or glucose, or had pH greater than 6.4 (Group I) and were assayed for osmolality, SG by urinometer and refractometer, and several other parameters, intercorrelations were calculated. Osmolality was used as the gold standard. In Group I, correlation of osmolality with N-Multistix SG (r = 0.66) was not significantly different (p greater than 0.1) from that found with SG determined by refractometer (r = 0.72), or urinometer (r = 0.60). The correlation coefficient for 45 N-Multistix-SG values determined by two staff members was 0.91. For the 127 Group II normal urines, the correlation coefficient between Multistix-SG and SG by refractometer was 0.85. Multistix-SG avoids the errors related to large molecules such as glucose and radiographic contrast media seen with SG determined by urinometer and refractometer (uncorrected for glucose) and was found to have comparable accuracy to these two methods when compared with osmolality, in patients with known renal disease or abnormal urinalysis.

Humans↗

Artificial gravity: head movements during short-radius centrifugation.

Short-radius centrifugation is a potential countermeasure to long-term weightlessness. Unfortunately, head movements in a rotating environment induce serious discomfort, non-compensatory vestibulo-ocular reflexes, and subjective illusions of body tilt. In two experiments we investigated the effects of pitch and yaw head movements in participants placed supine on a rotating bed with their head at the center of rotation, feet at the rim. The vast majority of participants experienced motion sickness, inappropriate vertical nystagmus and illusory tilt and roll as predicted by a semicircular canal model. However, a small but significant number of the 28 participants experienced tilt in the predicted plane but in the opposite direction. Heart rate was elevated following one-second duration head turns. Significant adaptation occurred following a series of head turns in the light. Vertical nystagmus, motion sickness and illusory tilt all decreased with adaptation. Consequences for artificial gravity produced by short-radius centrifuges as a countermeasure are discussed. Grant numbers: NCC 9-58.

Adaptation, Physiological↗

Neurobiology of fish under altered gravity conditions.

In vertebrates (including man), an altered gravitational environment such as weightlessness can induce malfunction of the inner ear, based on an irregular dislocation of the otoliths from the corresponding sensory epithelia. This dislocation leads to an illusionary tilt, since the otolithic inputs are not in register with other sensory organs. This results in an intersensory conflict. Vertebrates in orbit therefore face severe orientation problems. In humans, the intersensory conflict may additionally lead to a malaise, commonly referred to as space motion sickness (SMS). During the first days in weightlessness, the orientation problems (and SMS) disappear, since the brain develops a new compensatory interpretation of the available sensory data. The present review reports the neurobiological responses-particularly in fish-observed at altered gravitational states, concerning behaviour and neuroplastic reactivities. Recent investigations employing microgravity (spaceflight, parabolic aircraft flights, clinostat) and hyper-gravity (laboratory centrifuges as ground based research tools) yielded clues and insights into the understanding of the respective basic phenomena. The possible sources of human space sickness (a kinetosis) and of the space adaptation syndrome (when a sensory reinterpretation of gravitational and visual cues takes place) are particularly highlighted with regard to the functional significance of bilaterally asymmetric otoliths (weight, size).

Animals↗

Adaptation in a rotating artificial gravity environment.

The centripetal force generated by a rotating space vehicle is a potential source of artificial gravity. Minimizing the cost of such a vehicle dictates using the smallest radius and highest rotation rate possible, but head movements made at high rotation rates generate disorienting, nauseogenic cross-coupled semicircular canal stimulation. Early studies suggested 3 or 4 rpm as the highest rate at which humans could adapt to this vestibular stimulus. These studies neglected the concomitant Coriolis force actions on the head/neck system. We assessed non-vestibular Coriolis effects by measuring arm and leg movements made in the center of a rotating room turning at 10 rpm and found that movement endpoints and trajectories are initially deviated; however, subjects readily adapt with 10-20 additional movements, even without seeing their errors. Equilibrium point theories of motor control errantly predict that Coriolis forces will not cause movement endpoint errors so that subjects will not have to adapt their reaching movements during rotation. Adaptation of movement trajectory acquired during Coriolis force perturbations of one arm transfers to the unexposed arm but there is no intermanual transfer of endpoint adaptation indicating that neuromotor representations of movement endpoint and trajectory are separable and can adapt independently, also contradictory to equilibrium point theories. Touching a surface at the end of reaching movements is required for complete endpoint adaptation in darkness but trajectory adapts completely with or without terminal contact. We have also made the first kinematic measurements of unconstrained head movements during rotation, these movements show rapid adaptation to Coriolis force perturbations. Our results point to methods for achieving full compensation for rotation up to 10 rpm.

Adaptation, Physiological↗

A relationship between urine specific gravity and hyponatremia in hospitalized schizophrenic patients.

The relationship between urine specific gravity (USG) and the incidence of polydipsia/hyponatremia in hospitalized schizophrenic patients was examined. On the basis of five independent USG values, we identified 49% (35 out of 72) of male and 22% (8 out of 37) of female patients with hyposthenuria (mean USG < or = 1.008). Review of 2 years of records of routine laboratory examinations identified 12 males with hyponatremia, 11 males with borderline hyponatremia, and 49 males with normal values; among female patients, only one patient with hyponatremia and three patients with borderline values were identified. A significantly lower mean USG (1.003 +/- 0.001) for the male patients with hyponatremia compared with the male patients with normal serum sodium data (1.011 +/- 0.005) was observed. All of the male patients with hyponatremia, as well as 18 of the 49 male patients with normal serum sodium values, exhibited hyposthenuria. The USG values of all the patients with hyponatremia were consistently quite low (< or = 1.005), whereas 14 of the 18 normonatremia/hyposthenuria patients had normal USG values (i.e. > or = 1.009) in at least one of five determinations; the mean USG values for most (16 of 18) of the normonatremia/hyposthenuria patients ranged from 1.006 to 1.008. Thus, many hospitalized schizophrenic patients exhibit hyposthenuria of varying degrees, but consistently low USG values are most suggestive of the risk of polydipsia/hyponatremia.

Adult↗

Gravity sensing in moss protonemata.

Moss protonemata are a valuable system for studying gravitropism because both sensing and upward curvature (oriented tip growth) take place in the same cell. We review existing evidence, especially for Ceratodon purpureus, that addresses whether the mass that functions in sensing is that of amyloplasts that sediment. Recent experiments show that gravitropism can take place in media that are denser than the apical cell. This indicates that gravity sensing relies on an intracellular mass rather than that of the entire cell and provides further support for the starch-statolith hypothesis of sensing. Possible mechanisms for how amyloplast mass functions in sensing and transduction are discussed.

Bryopsida↗

Otolith formation in a mutant Medaka with a deficiency in gravity-sensing.

Mutant Medaka ha exhibit spontaneous mutation that is characterized by frequent inhibition or perturbation in the formation of utricular otoliths and/or semicircular canals. Three major features of otolith morphogenesis were observed in ha strain: 1) The initial appearance of otoliths was delayed, mispositioned, and malformed compared to normal embryos. 2) No utricular otoliths appeared on macula of any ha fry just after hatching. A symmetric state of otoliths was seen only when saccular otoliths were situated on macula in both inner ears. 3) In some fry, formation of utricular otoliths was observed in their later development. However, no new utricular otoliths appeared after fish were seventy or more days old after hatching. These observations show that otolith morphogenesis in ha is very different from that of wild-type. In this study, we classified adult ha into four different phenotypes using the existence or absence of utricular otoliths as our criteria. We concluded that dysfunction of utricular otoliths and semicircular canals cause a defect that affects the gravity-sensing abilities of medaka ha.

Animals↗

Cellular responses to gravity: extracellular, intracellular and in-between.

Our understanding of gravitational effects (inertial effects in the vicinity of 1 x g) on cells has matured to a stage at which it is possible to define, on the basis of experimental evidence, extracellular effects on small cells and intracellular effects on eukaryotic gravisensing cells. Yet undetermined is the nature of response, if any, of those classes of cells that are not governed solely by extracellular physical events (as are prokaryotes) and are devoid of obvious mechanical devices for sensing inertial forces (such as those possessed by certain plant cells and sensory cells of animals). This "in-between" class of cells needs to be understood on the basis of the combination of intracellular and extracellular gravity-dependent processes that govern experimentally-measurable variables that are relevant to the cell's responses to modified inertial forces. The forces that certain cell types generate or respond to are therefore compared to those imposed by approximately 1 x g in the context of cytoskeletal action and symmetry-breaking pathways.

Animals↗

Involvement of the second messenger cAMP in gravity-signal transduction in Physarum.

The aim of the investigation was to clarify, whether cellular signal processing following graviperception involves second messenger pathways. The test object was a most gravisensitive free-living ameboid cell, the myxomycete (acellular slime mold) Physarum polycephalum. It was demonstrated that the motor response is related to acceleration-dependent changes in the levels of the cellular second messenger cyclic adenosine monophosphate (cAMP). Rotating Physarum plasmodia in the gravity field of the Earth about a horizontal axis increased their cAMP concentration. Depriving the cells for a few days of the acceleration stimulus (near weightlessness in a space experiment on STS-69) slightly lowered plasmodial cAMP levels. Thus, the results provide first indications that the acceleration-stimulus signal transduction chain of Physarum uses an ubiquitous second messenger pathway.

Animals↗

On inappropriately used neuronal circuits as a possible basis of the "loop-swimming" behaviour of fish under reduced gravity: a theoretical study.

One hypothesis for the explanation of the so-called "loop-swimming" behaviour in fish when being subjected to reduced gravity assumes that the activities of the differently weighted otoliths of the two labyrinths are well compensated on ground but that a functional asymmetry is induced in weightlessness, resulting in a tonus asymmetry of the body and by this generating the "loop-swimming" behaviour. The basis of this abnormal behaviour has to be searched for in the central nervous system (cns), where the signal-transduction from the inner ear- related signal internalisation to the signal response takes place. Circuits within the CNS of fish, that could possibly generate the "loop-swimming", might be as follows: An asymmetric activation of vestibulospinal circuits would directly result in a tonus asymmetry of the body. An asymmetric activation of the oculomotor nucleus would generate an asymmetrical rotation of the eyes. This would cause in its turn asymmetric images on the two retinas, which were forwarded to the diencephalic accessory optic system (AOS). It is the task of the AOS to stabilize retinal images, thereby involving the cerebellum, which is the main integration center for sensory and motor modalities. With this, the cerebellar output would generate a tonus asymmetry of the body in order to make the body of the fish follow its eyes. Such movements (especially when assuming an open loop control) would end up in the aforementioned "loop-swimming" behaviour.

Animals↗

Oxygen cost of dynamic leg exercise on a cycle ergometer: effects of gravity acceleration.

A model of the metabolic internal power (Eint) during cycling, which includes the gravity acceleration (ag) as a variable, is presented. This model predicts that Eint is minimal in microgravity (0 g; g=9. 81 m s-2), and increases linearly with ag, whence the hypothesis that the oxygen uptake (VO2) during cycling depends on ag. Repeated VO2 measurements during steady-state exercise at 50, 75 and 100 W on the cycle ergometer, performed in space (0 g) and on Earth (1 g) on two subjects, validated the model. VO2 was determined from the time course of decreasing O2 fraction during rebreathing. The gas volume during rebreathing was determined by the dilution principle, using an insoluble inert gas (SF6). Average VO2 for subject 1 at each power was 0.99, 1.21 and 1.52 L min-1 at 1 g (n=3) and 0.91, 1.13 and 1.32 L min-1 at 0 g (n=5). For subject 2 it was 0.90, 1.12 and 1. 42 L min-1 at 1 g, and 0.76, 0.98 and 1.21 L min-1 at 0 g. These values corresponded to those predicted from the model. Although resting VO2 was lower at 0 g than at 1 g, the net (total minus resting) exercise VO2 was still smaller at 0 g than at 1 g. This difference reflects the lower Eint at 0 g.

Adult↗

A dipstick protein and specific gravity algorithm accurately predicts pathological proteinuria.

BACKGROUND: The proper strategy to screen for early chronic kidney disease is debatable, but protein-creatinine ratio from a random urine sample (UPC) commonly is used. The purpose is to determine whether dipstick data effectively identify patients with increased UPC ratios. Because urine concentration affects proteinuria interpretation, we hypothesized that dipstick protein (DSP) and specific gravity (SG) are sufficient for screening. METHODS: A hospital laboratory database was searched for urine samples simultaneously assayed for UPC ratio, DSP, and SG (n = 2,098). A random 70% of the cohort was used to generate a development model, which was validated in the remaining 30%. Samples were stratified according to DSP and SG values. A DSP versus SG matrix was created, and each sample was allocated to a discrete DSP-SG category. Proportions of samples with overt (UPC ratio > or = 500 mg/g) and nephrotic-range proteinuria (UPC ratio > or = 3,000 mg/g) were calculated for all 40 cells. RESULTS: Optimum correlations between DSP-SG cells and UPC ratios were determined for the development model, yielding 97.0% negative predictive value (NPV) for a UPC ratio of 500 mg/g or less and 97.5% positive predictive value (PPV) for a UPC ratio of 500 mg/g or greater. NPV for a UPC ratio of 3,000 mg/g or less was 99.7%. Application of the model to the validation sample showed a 96.5% NPV for a UPC ratio of 500 mg/g or less, 99.4% PPV for a UPC ratio of 500 mg/g or greater, and 99.0% NPV for a UPC ratio of 3,000 mg/g or less. CONCLUSION: DSP and SG values effectively identify patients requiring proteinuria quantification by means of UPC ratio. A Web-based tool was developed that allows DSP and SG value entry and provides a recommendation regarding the need for proteinuria quantification.

Biomarkers↗

Consistently high urine specific gravity in adolescent American football players and the impact of an acute drinking strategy.

The purpose of this study was to determine whether high-school football players showed risks of fluid deficits during two-a-day training (Part 1), and whether implementing a drinking strategy could acutely improve the markers of hydration (Part 2). In Part 1, pre-training urine specific gravity (USG) and pre- and post-training body weight were measured at the morning session for 5 consecutive days of two-a-day practices to monitor the hydration status of 13 varsity players. The mean pre-training body weight was consistently lower (mean decrease of 0.5 kg, p<0.05) following the first day of measurement. Pre-training USG values remained consistently high each day (range for daily means: 1.022+/-0.003 to 1.024+/-0.005). Part 2 consisted of assessing hydration status in 46 varsity and junior varsity players prior to morning training during two-a-day training before and following implementing a drinking strategy. In association with the strategy, mean body weight increased 0.5 kg (p<0.01) and mean USG decreased from 1.021 to 1.016 (p<0.01) following the drinking protocol. The slight decline in body weight and consistently high USG (Part 1) suggested that standard fluid replacement strategies were less than optimal for a majority of the players. Implementing a drinking strategy appeared to improve hydration status based on changes in body weight and USG (Part 2).

Adolescent↗