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c-fos and HSP70 gene expression in rat brains in high gravitation-induced cerebral ischemia.

Previous studies have shown that brief exposures of rodents to high gravitational forces (+Gz) in a specifically designed centrifuge cause global cerebral ischemia. In the present study, the effect of +Gz exposure to +22.5Gz for 15 to 60 s on c-fos and HSP70 gene expression was examined. Northern and RT-PCR analyses to total RNA isolated from brains of rats in different post-exposure times revealed a significant, time-dependent increase in the c-fos mRNA level which returned to near normal by 180 min. The HSP70 mRNA level was increased two-fold at 30 min post exposure, and remained elevated until 180 min. The transient stimulation of c-fos and HSP70 gene expression should serve as useful biomarkers for hypergravic stress on the brain. The present results should aid in design of future experiments in our understanding of the pathophysiology of the high +Gz challenges.

Animals↗

Separation of living red blood cells by gravitational field-flow fractionation.

The field-flow fractionation technique, using the earth's gravitational field, has been applied to peripheral blood cell populations. A more or less symmetrical, gaussian-like, elution peak is generally observed for the red cell population. The bimodal cell population obtained after a massive transfusion is shown to result in a shoulder on the red blood cell elution profile. In one case where a similar shouldering peak was obtained from a non-transfused donor, the existence of an immunological double population has been demonstrated. This suggests that field-flow fractionation has some potential for complementary biomedical diagnosis.

Cell Separation↗

Investigation of red blood cell fractionation by gravitational field-flow fractionation.

Gravitational field-flow fractionation is used for the separation of particles according to their sizes in the range 1-100 microns: larger particles elute before smaller ones. This phenomenon can be explained as a result of the steric exclusion of the particles from the vicinity of the channel walls, and/or hydrodynamic effects supposedly associated with the inertia of the liquid. The method was used for the investigation of red blood cells. The dependence of the retention ratio on the flow-rate, sample volume, concentration of blood and relaxation time was studied. Analysis of fifteen individual fractions by Coulter counter and reinjection of three other fractions were studied in order to verify fractionation of red blood cells.

Cell Separation↗

Monitoring of an experimental red blood cell pathology with gravitational field-flow fractionation.

Gravitational field-flow fractionation is a simple method suitable for the separation of micrometre-size particles, for example the red blood cells (RBC). The variation in the composition of the RBC population in a rabbit during an experimental reversible anaemia induced by phenylhydrazine was studied. Blood samples taken at different stages of the anemia showed differences in the retention and shape of the elution profiles. Microscopic observations of original RBC samples and fractions collected at the outlet of the fractionation channel make a description of the RBC population possible. The three species observed were the normal RBC, the newly produced reticulocytes, and the blood cells containing Heinz bodies (intracellular haemoglobin precipitates). A decrease of the normal RBC from 96 to 1% was observed over five days. The production of reticulocytes in bone marrow is stimulated by the anaemia and increases in percentage after the second day of the anaemia (from 1 to 16%). RBC with Heinz bodies, which appear on the third day, were also studied. Granulometric studies were performed on the RBC sampled from the rabbit each day as well as on some fractions eluted by field-flow fractionation. Reinjection procedures of some cell subpopulations of known size distribution were also performed. The relaxation process of these cells was then studied to approximate their density properties. It was observed that RBC of different density but of the same average size were selectively eluted, as were cells of equivalent density but of different size. Injection of the cells at different stop-flow times enabled the study of the relaxation process on the elution profiles. The results, compared with systematic microscopic observation and size analysis, permit the description of modifications in the RBC composition as well as the purification of subpopulations at each stage of the anaemia. The correlation observed between the fractionation profiles and the progress or the regression of the anaemia opens a new field in the analytical monitoring of this type of pathology.

Anemia, Hemolytic↗

Biochemical and functional differences in rat liver mitochondrial subpopulations obtained at different gravitational forces.

Previous studies have reported that liver mitochondria may be fractionated into different subpopulations. However, no careful studies have been performed to exclude mitochondrial damage and to investigate more thoroughly the possible biochemical differences existing between the subpopulations. In this study, we analysed the integrity and the biochemical properties of rat liver mitochondria. Mitochondrial fractions were obtained by differential centrifugation at different gravitational forces: 1000 g (M1 fraction), 3000 g (M3 fraction) and 10,000 g (M10 fraction). The integrity of these organelles was checked by measuring citrate synthase activity both in the presence and absence of Triton X-100 detergent. Biochemical analyses included polarographic determination of cytochrome oxidase activity and respiratory parameters and spectrophotometric determination of cytochrome content. (1) The integrity of mitochondria was almost homogeneous between fractions (88.5, 80 and 78.3% in M1, M3 and M10 fractions, respectively). (2) The heaviest M1 fraction contains mitochondria which are on average twice as large as M3 and about three times as large as M10. (3) The M1 fraction exhibited the highest specific cytochrome oxidase activity (1040 +/- 20 n Atoms O/min x mg protein) and the highest respiratory rates (72 +/- 3 n Atoms O/min x mg protein and 526 +/- 45 n Atoms O/min x mg protein for States 4 and 3, respectively). Oxidative capacity and respiratory rates decreased as the size of the organelles decreased, reaching values of 1/5 and 1/14 in the M3 and M10 fractions as compared to the M1. (4) These changes are accompanied by a change in the respiratory control ratio (RCR), which varies from 7.3 in M1 to about 2.0 in M10. A similar trend was observed in cytochrome contents but the differences were not as great as cytochrome oxidase activity and State 3 respiration. These results, as a whole, show that a mitochondrial heterogeneity exists in rat liver cell. We suggest that the above-mentioned differences might represent steps of mitochondrial maturation. The maturation would be fundamentally based on the increase of efficiency of the mechanism for ATP synthesis.

Acid Phosphatase↗

Effects of gravitational and magnetic fields on transplanted neuroblastoma vascularity.

Transplanted neuroblastomas were subjected to altered neural influences and imposed gravitational and magnetic fields, either separately or in combination. In each case impaired vascularity, and greater extravasation of blood with associated tumor necrosis occurred in the experimental animals. The combination of both hypergravity and alternating magnetic field resulted in definite macroscopic but less marked microscopic changes than with either force alone. Neuroblastoma cells in vitro exposed to a magnetic field showed no definite growth changes, suggesting an effect of the field forces on the host. Exposure of a unicellular organism, not requiring a host for survival, to a similar magnetic field, did result in distinct cell division changes.

Animals↗

Valsalva and gravitational variability of the internal jugular vein and common femoral vein: ultrasound assessment.

PURPOSE: Central venous cannulation via the common femoral vein is an important starting point for many interventions. The purpose of this study was to determine the optimum conditions for cannulation of the femoral vein and to compare these with the relative changes in the internal jugular vein. METHODS: High-resolution 2D ultrasound was utilised to determine variability of the calibre of the femoral and internal jugular veins in 10 healthy subjects. Venous diameter was assessed during the Valsalva manoeuvre and in different degrees of the Trendelenburg position. RESULTS: The Valsalva manoeuvre significantly increased the size of the femoral and internal jugular veins. There was a relatively greater increase in femoral vein diameter when compared with the internal jugular vein of 40 and 29%, respectively. Changes in body inclination (Trendelenburg position) did not significantly alter the luminal diameter of the femoral vein. However, it significantly increased internal jugular vein diameter. CONCLUSIONS: Femoral vein cannulation is augmented by the Valsalva manoeuvre but not significantly altered by the gravitational position of the subject.

Adult↗

Analytical model of solute transport by unsteady unsaturated gravitational infiltration.

Penetration of reactive solute into a soil during a cycle of water infiltration and redistribution is investigated by deriving analytical closed form solutions for fluid flux, moisture content and contaminant concentration. The solution is developed for gravitational flow and advective transport and is applied to two scenarios of solute applications encountered in the applications: a finite pulse of solute dissolved in irrigation water and an instantaneous pulse broadcasted onto the soil surface. Through comparison to simulations of Richards' flow, capillary suction is shown to have contrasting effects on the upper and lower boundaries of the fluid pulse, speeding penetration of the wetting front and reducing the rate of drying. This leads to agreement between the analytical and numerical solutions for typical field and experimental conditions. The analytical solution is further incorporated into a stochastic column model of flow and transport to compute mean solute concentration in a heterogeneous field. An unusual phenomenon of plume contraction is observed at long times of solute propagation during the drying stage. The mean concentration profiles match those of the Monte-Carlo simulations for capillary length scales typical of sandy soils.

Gravitation↗

Optimization of experimental conditions for the separation of small and large starch granules by gravitational field-flow fractionation.

Separation of small and large barley starch granules by gravitational field-flow fractionation was investigated from the point of view of sample pre-treatment, amount of injected sample, and elution conditions. The sample pre-treatment study resulted in the conclusion that it is reasonable to soak the starch granules for at least 24 h prior to separation. The experiments with different amounts of injected sample show that it is possible to increase as well as decrease twofold the sample amount usually used without any change in retention ratios. The implementation of flow-rate gradients for elution of the starch granules reduced total separation time. However, the applied flow-rate gradients did not improve the resolution of peaks A and B compared with the generally used constant flow-rate. Thus, for barley starch granules, the constant flow-rates within the range from 0.8 to 1.0 ml/min seem to provide the best compromise of total separation time, peak resolution and instrumental expense.

Chemical Fractionation↗

Gravitational field-flow fractionation for the characterisation of active dry wine yeast.

Gravitational field-flow fractionation (GrFFF) is applied to the fractionation of active dry wine yeast. An experimental approach to the analysis of the effects that field variation by changing mobile phase composition and flow-rate have on the fractionation process of standard particles (polystyrene) was first developed to further obtain effective fractionation of wine yeast by GrFFF. Scanning electron microscopy and Coulter counter particle size measurements were used to monitor the fractionation extent and capabilities of GrFFF to describe the distribution of yeast cells populations.

Chemical Fractionation↗

Improved performance of gravitational field-flow fractionation for screening wine-making yeast varieties.

Performance of gravitational field-flow fractionation (GFFF) is improved here with respect to the ability to fractionate and distinguish different varieties of wine-making yeast from Saccharomyces cerevisiae. A new GFFF channel with non-polar walls has been employed to enhance fractionation selectivity and reproducibility. Since GFFF retention depends from first principles on particle size, Coulter counter measurements were performed in order to compare size distribution profiles with GFFF profiles. From such a comparison, GFFF was shown to be able to reveal differences in yeast cells other than size. This could make use of GFFF for screening different varieties of wine-making yeast towards future quality assessment procedures based on a possible correlation between yeast cell morphology indexes and quality indexes.

Fractionation, Field Flow↗

Gravitational effects on volume distribution in a model of partial and total liquid ventilation.

To estimate regional lung volume during ventilation with liquids (e. g. perfluorochemicals, PFC) we developed a multi-compartment mathematical model of a lung and thorax. The height of the fluid column and the fluid's density determine alveolar pressure (PA). The weight of thoracic contents above any given gravitational plane influences pleural pressure (PPL). Transpulmonary pressure (PTP=PA−PPL) and compliance of the lung and chest wall permit estimation of volumes. The results indicate the lung inflates almost uniformly during total liquid ventilation despite a substantial vertical PA gradient. Inflation uniformity is due to the offsetting vertical PPL gradient created by the added weight of the PFC and sustained by the relative rigidity of the chest wall. During partial liquid ventilation our model indicates that the combination of uniform PA with a large vertical gradient in PPL leads to a vertical PTP gradient and therefore relative over-inflation of the top of the lung. This effect increases with increasing PFC dose and with lung height.

Functional Residual Capacity↗

Gravitational sedimentation of flocculated waste activated sludge.

The sedimentation characteristics of flocculated wastewater sludge have not been satisfactorily explored using the non-destructive techniques, partially owing to the rather low solid content (ca. 1-2%) commonly noted in the biological sediments. This paper investigated, for the first time, the spatial-temporal gravitational settling characteristics of original and polyelectrolyte flocculated waste activated sludge using Computerized Axial Tomography Scanner. The waste activated sludge possessed a distinct settling characteristic from the kaolin slurries. The waste activated sludges settled more slowly and reached a lower solid fraction in the final sediment than the latter. Flocculation markedly enhanced the settleability of both sludges. Although the maximum achievable solid contents for the kaolin slurries were reduced, flocculation had little effects on the activated sludge. The purely plastic rheological model by Buscall and White (J Chem Soc Faraday Trans 1(83) (1987) 873) interpreted the consolidating sediment data, while the purely elastic model by Tiller and Leu (J. Chin. Inst. Chem. Eng. 11 (1980) 61) described the final equilibrated sediment. Flocculation produced lower yield stress during transient settling, thereby resulting in the more easily consolidated sludge than the original sample. Meanwhile, the flocculated activated sludge was stiffer in the final sediment than in the original sample. The data reported herein are valuable to the theories development for clarifier design and operation.

Flocculation↗

Invariant aspects of human locomotion in different gravitational environments.

Previous literature showed that walking gait follows the same mechanical paradigm, i.e. the straight/inverted pendulum, regardless the body size, the number of legs, and the amount of gravity acceleration. The Froude number, a dimensionless parameter originally designed to normalize the same (pendulum-like) motion in differently sized subjects, proved to be useful also in the comparison, within the same subject, of walking in heterogravity. In this paper the theory of dynamic similarity is tested by comparing the predictive power of the Froude number in terms of walking speed to previously published data on walking in hypogravity simulators. It is concluded that the Froude number is a good first predictor of the optimal walking speed and of the transition speed between walking and running in different gravitational conditions. According to the Froude number a dynamically similar walking speed on another planet can be calculated as [formula: see text] where V(Earth) is the reference speed on Earth.

Acceleration↗

Effects of gravitational changes on the bone system in vitro and in vivo.

Spaceflight data obtained on bone cells, rodents, and humans are beginning to shed light on the importance of gravitational loading on the skeletal system. The space environment is a relevant model to explore the bone cell response to minimal strains. However, whether there is a direct effect of gravity on the cell rather than changes related to lack of convection forces in cell cultures performed in microgravity is unknown. In vitro studies carried out using osteoblastic cell cultures in space show changes in cell shape, suggesting that cell attachment structures as well as cytoskeleton reorganization might be involved. Valuable information is expected from in vitro models of an increase or decrease in mechanical stress in order to identify the different pathways of mechanoreception and mechanotransduction in the osteoblastic lineage. Results obtained from both humans and rodents after spaceflights indicated that bone mass changes are site specific rather than evenly distributed throughout the skeleton, thus emphasizing the need to perform measurements at different bone sites: weight- and non-weight-bearing bones, and cancellous and cortical envelopes. Bone mass measurements and biochemical parameters of bone remodeling are currently under evaluation in cosmonauts. Histomorphometric studies of bones from rats after space missions of various periods provided the time course of the cancellous bone cellular events: transient increase in resorption and sustained decrease in bone formation. The underlying bone loss occurred first in weight-bearing bones and later in less weight-bearing bones. During the postflight period, time required to recover the lost bone was greater than the mission length. Thus, the postflight period deserves more attention than it is currently receiving. On earth, the rat tail-suspension model is currently used to mimic spaceflight-induced bone loss. Data from the model confirmed the impairment of osteoblastic activity and showed an alteration in osteoblast recruitment with skeletal unloading. However, this model needs to be further validated.

Adaptation, Physiological↗

Determination of viable yeast cells by gravitational field-flow fractionation with fluorescence detection.

Vinification processing is largely related to yeast performance and depends on the initial cell viability. To optimize the quality of wine fermentation, control of the yeast quality is mandatory. The present paper describes a new method using gravitational field flow fractionation (GrFFF) with fluorescence detection for the determination of yeast cell viability before the fermentation process. A GrFFF calibration procedure was developed using commercial yeast to prepare standards of viable cells and propidium iodide (PI) as fluorescent probe for nonviable cells. The suitability of the new method was tested with several commercial yeast strains with a g/L content ranging from 1 to 3. The validation of the method was performed by comparing GrFFF viability values with those obtained using Coulter counter and flow cytometry techniques.

Cell Separation↗

Use of fluorescent probes for determination of yeast cell viability by gravitational field-flow fractionation.

The quality of wine greatly depends on the features of the yeast used in its production, and yeast cell viability is one of the most important quality control issues to consider in this regard. In the first steps of winemaking, the use of a low-cost and simple methodology for monitoring the cell viability of yeast inoculates is of paramount importance. Gravitational field-flow fractionation is a useful technique for the determination of cell viability because it provides gentle experimental conditions, although the proper use of fluorophore probes as biomass indicators is required. In this paper the use of different fluorescent probes such as carboxyfluorescein diacetate (cFDA), calcein-AM, and SYTO-13 were considered as viability biomarkers. Calceina-AM allowed the establishment of a direct GrFFF method to determine cell viability, with a limit of detection of 5.0 x 10(4) viable cell/mL. SYTO-13 could be used as biomass indicator with a limit of detection of 3.5 x 10(4) total cells/mL. The suitability of the procedure was tested with three commercial yeast samples, and the results were compared with those obtained using standard techniques.

Calibration↗

Impact of weighting agents and sucrose on gravitational separation of beverage emulsions.

The influence of weighting agents and sucrose on gravitational separation in 1 wt % oil-in-water emulsions was studied by measuring changes in the intensity of backscattered light from the emulsions with height. Emulsions with different droplet densities were prepared by mixing weighting agents [brominated vegetable oil (BVO), ester gum (EG), damar gum (DG), or sucrose acetate isobutyrate (SAIB)] with soybean oil prior to homogenization. Sedimentation or creaming occurred when the droplet density was greater than or lower than the aqueous phase density, respectively. The weighting agent concentrations required to match the oil and aqueous phase densities were 25 wt % BVO, 55 wt % EG, 55 wt % DG, and 45 wt % SAIB. The efficiency of droplet reduction during homogenization also depended on weighting agent type (BVO > SAIB > DG, EG) due to differences in oil phase viscosity. The influence of sucrose (0-13 wt %) on the creaming stability of 1 wt % soybean oil-in-water emulsions was also examined. Sucrose increased the aqueous phase viscosity (retarding creaming) and increased the density contrast between droplets and aqueous phase (accelerating creaming). These two effects largely canceled one another so that the creaming stability was relatively insensitive to sucrose concentration.

Beverages↗