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Fibrinolysis of experimental subretinal haemorrhage without removal using tissue plasminogen activator.

AIMS/BACKGROUND: Human recombinant tissue plasminogen activator (rt-PA) fibrinolysis of subretinal haemorrhage with concomitant removal has been shown to reverse the natural history of photoreceptor degeneration in experimental subretinal haemorrhages if evacuated within 7 days. The aim of the study was to determine whether fibrinolysis of subretinal haemorrhage without concomitant removal would offer a simpler approach with similar photoreceptor sparing. METHODS: A neodymium YAG laser was used to create experimental subretinal haemorrhages beneath the holangiotic retina of the cat. Tissue plasminogen activator (10 micrograms/ml) was injected into 4 day old subretinal haemorrhages to evaluate its effect on altering the natural history of retinal degeneration. Light and electron microscopy were used to study the histopathological effect. RESULTS: The injection of rt-PA into large 4 day old subretinal haemorrhages without concomitant removal did not alter the natural history of retinal degeneration. In fact, a second focus of retinal degeneration occurred at a gravity dependent inferior site where the subretinal haemorrhages had migrated. CONCLUSIONS: There was no therapeutic benefit from the injection of rt-PA into subretinal haemorrhages without con-comitant removal in this cat model.

Animals↗

Mechanical adaptability of a sponge extracellular matrix: Evidence for cellular control of mesohyl stiffness in Chondrosia reniformis Nardo.

The marine sponge Chondrosia reniformis Nardo consists largely of a collagenous tissue, the mesohyl, which confers a cartilaginous consistency on the whole animal. This investigation was prompted by the incidental observation that, despite a paucity of potentially contractile elements in the mesohyl, intact C. reniformis stiffen noticeably when touched. By measuring the deflection under gravity of beam-shaped tissue samples, it was demonstrated that the flexural stiffness of the mesohyl is altered by treatments that influence cellular activities, including [Ca2+] manipulation, inorganic and organic calcium channel-blockers and cell membrane disrupters, and that it is also sensitive to extracts of C. reniformis tissue that have been repeatedly frozen then thawed. Since the membrane disrupters and tissue extracts cause marked stiffening of mesohyl samples, it is hypothesised that cells in the mesohyl store a stiffening factor and that the physiologically controlled release of this factor is responsible for the touch-induced stiffening of intact animals.

Animals↗

Effects of exposure to space flight on endocrine regulations in experimental animals.

This minireview summarizes the results of the observations on changes in endocrine functions of rats exposed to space flights for various periods. The results found after space flights are compared with those obtained from rats in acute or repeated restrain stress. A slight increase of plasma catecholamine levels was observed in rats after space flight of longer duration (>14 days), but no changes in catecholamine content in the activity of catecholamine synthesizing enzymes were noted in adrenal medulla and in hypothalamus. The norepinephrine content was, however, decreased in several nuclei selected from hypothalamus of flight rats. Plasma corticosterone levels were increased after space flight and morphological examination of pituitary showed elevated activity of corticotrophs. However, the plasma levels of ACTH were not increased in rats 6 hours after space flight. These changes in plasma hormone levels affected the activity of enzymes involved in metabolism of amino acids in liver and lipolysis in adipose tissue. The plasma levels of testosterone and triiodothyronine were diminished after space flight suggesting the suppression of the thyroid and gonadal activity. Increase of plasma insulin and glucose levels were found in rats after space flight, but the glucagon values were not changed. Comparing these results from flight rats with the animals exposed to acute or repeated stress indicate that long stay in microgravity do not represent very intensive stressogenic stimulus for adrenocortical and sympatho-adrenomedullar systems, and hormone alterations observed after space flight may be due to acute gravitational stress resulting from a return to Earth gravity. Therefore further studies including the inflight animal experiments on a board of International Space Station are necessary for elucidation of the effects of microgravity on endocrine functions.

Animals↗

[Pellet guns: a persistent threat to eyes].

Following a recent case of bilateral perforating ocular trauma by multiple pellets, the authors reviewed pellet traumatisms treated in the Emergency Department and operated on in the Ophthalmology Department of the Villeneuve-St-Georges Hospital over the past 15 years. At close range, pellet weapon shootings can generate lid, conjunctiva, and powder cornea tattoos; in these cases, the lesions are often unilateral. Shooting at longer range does not result in corneodermic tattoos, but because pellets scatter, binocular lesions frequently occur. The different lesions caused by these nonmagnetic foreign bodies are often very serious. An early, even very early, traumatic cataract is sometimes very difficult to treat in these polytraumatized eyes. The visual prognosis is more reserved, as the impact zones often involve the posterior pole and the macular area. The best recovered visual acuity is 30-35/20, with visual field alterations, but the injury to the eyeball by multiple pellets can generate globe atrophy. The authors stress the gravity of these ocular traumas, fortunately rarer but not completely eradicated since the law of May 6, 1995, which restricted the possession of these pellet guns.

Adolescent↗

[Changes in chronic pancreatitis. Functional, structural and histological studies].

Pancreatic function, structural changes in comparison with morphologic (histological) ones were studied in 22 patients suffering from chronic pancreatitis. Gravity of functional and structural damages--apart from minor differences--showed close cinnection with the severity of tissue alterations. In half of the cases, combination of tissue signs of obstructive and calcificating pancreatitis were observed, so it is considered gratuitous to separate sharply the two forms of chronic pancreatitis.

Adult↗

Cellular adhesion in neoplastic and syngeneic normal cells under altered gravitational conditions.

The major objective of several experiments performed in space in the last 15 years was to establish whether single cells are sensitive to gravity. It was found in certain cells that reduced gravity leads to profound changes of a number of physiological functions like genetic expression, cell proliferation, signal transduction and cytoskeleton structure. In cell biology studies microgravity can be simulated on Earth in the clinostat. Nearly all data on experiments in the clinostat are related to cells cultured in suspension and, therefore, to adhesion-independent cells. In contrast, several biological phenomena as neoplastic transformation, cell differentiation, in-vitro cellular aging, contact inhibition and cellular adhesion require mainly cellular systems that are adhesion-dependent. The purpose of this work was: a) to study the behaviour of two rat cell strains (neoplastic SGS/4A and syngeneic fibroblasts FG) in order to test whether adhesion-dependent cells are suitable for clinorotation and b) to investigate cell-cell and cell-substratum adhesion in these cells kept under simulated low-g in the fast rotating clinostat and in hypergravity at l0g in the centrifuge.

Animals↗

Body fluid alterations during head-down bed rest in men at moderate altitude.

To determine the effects of hypoxia on fluid balance responses to simulated zero-gravity, measurements were made in six subjects (acclimatized to 5,400 ft; 1,646 m) before and during -5 degrees continuous head-down bed rest (HDBR) over 8 d at 10,678 ft. The same subjects were studied again at this altitude without HDBR as a control (CON) using a cross-over design. During this time, they maintained normal upright day-time activities, sleeping in the horizontal position at night. Fluid balance changes during HDBR in hypoxia were more pronounced than similar measurements previously reported from HDBR studies at sea level. Plasma volume loss (-19% on day 6) was slightly greater and the diuresis and natriuresis were doubled in magnitude as compared to previous studies in normoxia and sustained for 4 d during hypoxia. These changes were associated with an immediate, but transient rise in plasma atrial natriuretic peptide (ANP) to day 4 of 140% in HDBR and 41% in CON (p < 0.005), followed by a decline towards baseline. Differences were less striking between HDBR and CON for plasma antidiuretic hormone and aldosterone, which were transiently reduced by HDBR. Plasma catecholamines showed a similar pattern to ANP (+122%) in both HDBR and CON, suggesting that elevated ANP and catecholamines together accounted for the enhanced fluid shifts with HDBR during hypoxia.

Adult↗

Medical considerations for extending human presence in space.

The prospects for extending the length of time that humans can safely remain in space depend partly on resolution of a number of medical issues. Physiologic effects of weightlessness that may affect health during flight include loss of body fluid, functional alterations in the cardiovascular system, loss of red blood cells and bone mineral, compromised immune system function, and neurosensory disturbances. Some of the physiologic adaptations to weightlessness contribute to difficulties with readaptation to Earth's gravity. These include cardiovascular deconditioning and loss of body fluids and electrolytes; red blood cell mass; muscle mass, strength, and endurance; and bone mineral. Potentially harmful factors in space flight that are not related to weightlessness include radiation, altered circadian rhythms and rest/work cycles, and the closed, isolated environment of the spacecraft. There is no evidence that space flight has long-term effects on humans, except that bone mass lost during flight may not be replaced, and radiation damage is cumulative. However, the number of people who have spent several months or longer in space is still small. Only carefully-planned experiments in space preceded by thorough ground-based studies can provide the information needed to increase the amount of time humans can safely spend in space.

Adaptation, Physiological↗

Laboratory outreach: student assessment of flow cytometer fluidics in zero gravity.

Due to the the clinical utility of the flow cytometer, the National Aeronautics and Space Administration (NASA) is interested in the design of a space flight-compatible cytometer for use on long-duration space missions. Because fluid behavior is altered dramatically during space flight, it was deemed necessary to validate the principles of hydrodynamic focusing and laminar flow (cytometer fluidics) in a true microgravity environment. An experiment to validate these properties was conducted by 12 students from Sweetwater High School (Sweetwater, TX) participating in the NASA Reduced Gravity Student Flight Opportunity, Class of 2000. This program allows high school students to gain scientific experience by conducting an experiment on the NASA KC-135 zero gravity laboratory aircraft. The KC-135 creates actual zero-gravity conditions in 30-second intervals by flying a highly inclined parabolic flight path. The experiment was designed by their mentor in the program, the Johnson Space Center's flow cytometrist Brian Crucian, PhD, MT(ASCP). The students performed the experiment, with the mentor, onboard the NASA zero-gravity research aircraft in April 2000.

Aircraft↗

Simulated microgravity impairs leukemic cell survival through altering VEGFR-2/VEGF-A signaling pathway.

Motile cells capable of undergoing transendothelial migration, such as hematopoietic and leukemic cells, have been shown to sense and respond to a decrease in their surrounding gravity. In this study, we investigated the effects of microgravity on human leukemic cell proliferation and expression of receptors that control cell survival, such as the tyrosine kinase vascular endothelial growth factor receptor-2 (VEGFR-2). VEGFR-2 is shuttled between the nucleus and membrane, and through an autocrine activation of its ligand, VEGF-A, conveys signals that control cell survival. Autocrine or paracrine stimulation of VEGFR-2 facilitates localization of this receptor from the membrane to the nucleus--a process that results in increased survival of the leukemic cells. Here, we provide evidence that the mechanical forces altered by simulated microgravity localize and maintain VEGFR-2 in the membrane, and also block VEGF-A expression. This interferes with the shuttling of VEGFR-2 to the nucleus, resulting in a decrease in signaling and enhanced leukemic cell death. These data suggest that microgravity modulates cell survival through altering the cellular trafficking and activation state of tyrosine kinase receptors. This study has potential implications for understanding the regulation of receptor biology in pathophysiology, particularly VEGFR trafficking, thereby providing for the development of appropriate therapeutic strategies to abrogate intracrine stimulation triggered by VEGFR internalization.

Cell Proliferation↗

Vestibular adaptation to space in monkeys.

Otolith-induced eye movements of rhesus monkeys were studied before and after the 1989 COSMOS 2044 and the 1992 to 1993 COSMOS 2229 flights. Two animals flew in each mission for approximately 2 weeks. After flight, spatial orientation of the angular vestibulo-ocular reflex was altered. In one animal the time constant of postrotatory nystagmus, which had been shortened by head tilts with regard to gravity before flight, was unaffected by the same head tilts after flight. In another animal, eye velocity, which tended to align with a gravitational axis before flight, moved toward a body axis after flight. This shift of orientation disappeared by 7 days after landing. After flight, the magnitude of compensatory ocular counter-rolling was reduced by about 70% in both dynamic and static tilts. Modulation in vergence in response to naso-occipital linear acceleration during off-vertical axis rotation was reduced by more than 50%. These changes persisted for 11 days after recovery. An up and down asymmetry of vertical nystagmus was diminished for 7 days. Gains of the semicircular canal-induced horizontal and vertical angular vestibulo-ocular reflexes were unaffected in both flights, but the gain of the roll angular vestibulo-ocular reflex was decreased. These data indicate that there are short- and long-term changes in otolith-induced eye movements after adaptation to microgravity. These experiments also demonstrate the unique value of the monkey as a model for studying effects of vestibular adaptation in space. Eye movements can be measured in three dimensions in response to controlled vestibular and visual stimulation, and the results are directly applicable to human beings. Studies in monkeys to determine how otolith afferent input and central processing is altered by adaptation to microgravity should be an essential component of future space-related research.

Adaptation, Physiological↗

A two-person technique for epidural needle placement and medication infusion.

Traditional techniques of epidural needle placement have not proved satisfactory in patients who have had a previous laminectomy and may have epiduroarachnoiditis. A two-person technique was developed to: 1) simplify performance of difficult, epidural needle placement; 2) allow both resident and staff a simultaneous "hands-on" teaching experience; 3) allow a painless method of medication administration in patients with altered epidural anatomy; and 4) allow a simple method for determination of epidural space pressures. A glass syringe and extension tubing filled with saline are connected to the epidural needle. The needle is advanced by the operator while the pressure is monitored by the assistant. Once in the epidural space, the extension tubing is held vertically to act as a manometer. The column of saline falls and fluctuates with heart beat, providing verification of correct needle placement. Medication is placed in the barrel of the syringe and allowed to infuse by gravity. The two-person technique has achieved both patient and resident acceptance, and allows a painless method of medication administration in patients with altered epidural space anatomy.

Clinical Competence↗

Skylab experiment results: hematology studies.

These studies were designed and coordinated to evaluate specific aspects of man's immunologic and hematologic systems which might be altered by or respond to the space flight environment. The biochemical functions investigated included cytogenetic damage to blood cells, immune resistance to disease, regulation of plasma and red cell volumes, metabolic processes of the red blood cell, and physical chemical aspects of red blood cell functions. Only minor changes were observed in the functional capacity of erythrocytes as determined by measuring the concentrations of selected intracellular enzymes and metabolites. Tests of red cell osmotic regulation indicated some elevation in the activity of the metabolic dependent Na-K pump, with no significant alterations in the cellular Na and K concentrations or osmotic fragility. A transient shift in red cell specific-gravity profile was observed on recovery, possibly related to changes in cellular water content. Measurements of hemoconcentration (hematocrit, hemoglobin concentration, red cell count) indicated significant fluctuations postflight, reflecting observed changes in red cell mass and plasma volume. There was no apparent reticulocytosis during the 18 days following the first manned Skylab mission in spite of a significant loss in red cell mass. However, the reticulocyte count and index did increase significantly 5 to 7 days after completion of the second, longer duration, flight. There were no significant changes in either the while blood cell count or differential. However, the capacity of lymphocytes to respond to an in vitro mitogenic challenge was repressed postflight, and appeared to be related to mission duration. The cause of this repression is unknown at this time. Only minor differences were observed in plasma protein patterns. In the second mission there were changes in the proteins involved in the coagulation process which suggested a hypercoagulative condition.

Aerospace Medicine↗

Backtransport of dialysate solutes during in vitro continuous arteriovenous hemodialysis.

Simulated continuous arteriovenous hemodialysis (CAVHD) was performed with Renaflo anisotropic polysulfone capillary hemofilters and Biospal homogeneous PAN parallel-plate hemofilters. Inulin and glucose were present in the dialysis solution. Plasma and dialysis solution flow rates were varied as was the delivery of dialysis solution by gravity or pump. The presence of dialysis solution did not affect the components of transmembrane pressure and did not alter ultrafiltration rate. Inulin and glucose backtransported into the plasma perfusate during CAVHD with the Biospal but not with the Renaflo filters. Dialysis solution will need to be sterile and pyrogen-free even in the low-flow, low-pressure system of CAVHD.

Dialysis Solutions↗

Weightlessness experiments on Biosatellite II.

Four experiments in the aft compartment of Biosatellite II investigated the broad question of the effect of nearly zero gravity on the development, morphology and metabolism of plants and animals. The fertilization and development of the egg of a vertebrate (the frog, Rana pipiens), the feeding and growth of a protozoan (the giant amoeba, Pelomyxa carolinensis), the orientation of leaves and petioles of a young dicotyledon (pepper plants, Capsicum annuum) and the morphogenesis, orientation, histochemistry and biochemistry of a monocotyledon seedling (wheat, Triticum vulgare) gave a broad scope. All are known to have specific responses to normal gravity and changes in them might be expected to reflect the effects of orbital flight on living organisms. No differences in development of the frog eggs could be detected. Unfortunately, the 3 1/2 hour delay in launch allowed the first cleavage (the stage most sensitive to inversion) to appear before launch. Although the orbited embryos were somewhat slower to reach certain stages of development, recovered embryos developed just as did the controls. The amoebae fed normally while in orbit, and specimens fixed in orbit retained the ordinary heteropodal shape. Growth rates of orbited amoebae, both fed and starved, were slower than controls following reentry and recovery procedures. In continuous-fed organisms there was little or no effect of flight detectable in growth rate or actual number of divisions. Electron micrographs showed no abnormalities and few differences between flight and control organisms. The pepper plants were photographed in orbit at ten-minute intervals, as were the clinostat and erect controls. The subsequent measurement of photographs showed that in the orbited plants all leaves showed epinasty, the interaxial angle decreasing by 20-60 degrees C. Plants on the horizontal clinostat behaved comparably, but recovered more rapidly than orbited plants when returned to the normal erect position. Although the maximum age of wheat seedlings was only 65 hours, coleoptile and root growth rates during that time had not been significantly altered by flight or by slow rotation on a horizontal clinostat. There was some evidence that growth was accelerated after normal gravity was restored. The orientation of coleoptiles and of primary and lateral roots of orbited plants varied significantly from the normal erect seedlings but was almost identical with that of clinostat plants. The Periodic-Acid-Schiff technique on sectioned material showed starch grains at the bottom of cells of erect control coleoptile and root tips, while in orbited and clinostated plants the grains were located more or less at random. Histochemical differences between clinostat and orbited tissues are apparent however. Peroxidase localization varied and its activity was higher in both clinostat and orbited tissues; five other enzymes studied biochemically showed no differences. These experiments all suggest that there is no deleterious effect on living organisms or their activities from short-term weightlessness. Several results indicate that the horizontal clinostat may simulate the weightless state effectively here on Earth.

Amoeba↗

The effect of hypoxia on traumatic head injury in rats: alterations in neurologic function, brain edema, and cerebral blood flow.

We evaluated the effects of early posttraumatic hypoxia on neurologic function, magnetic resonance images (MRI), brain tissue specific gravities, and cerebral blood flow (CBF) in head-injured rats. By itself, an hypoxic insult (PaO2 40 mm Hg for 30 min) had little effect on any measure of cerebral function. After temporal fluid-percussion impact injury, however, hypoxia significantly increased morbidity. Of rats subjected to impact (4.9 +/- 0.3 atm) plus hypoxia, 71% had motor weakness contralateral to the impact side 24 h after injury, while only 29% of rats subjected to impact alone had demonstrable weakness (p less than 0.05). Lesions observed on MR images 24 h after injury were restricted to the impact site in rats with impact injury alone, but extensive areas with longer T1 relaxation times were observed throughout the ipsilateral cortex in rats with impact injury and hypoxic insult. Brain tissue specific gravity measurements indicated that much more widespread and severe edema developed in rats with impact injury and hypoxia. [14C]Iodoantipyrine autoradiography performed 24 h after injury showed that there was extensive hypoperfusion of the entire ipsilateral cortex in rats with impact injury and hypoxia. These results show that large areas of impact-injured brain are extremely vulnerable to secondary insults that can irreparably damage neural tissue, and provide experimental evidence for the observed adverse effects of hypoxia on outcome after human head injury.

Animals↗

Mapping the functional roles of cap cells in the response of Arabidopsis primary roots to gravity.

The cap is widely accepted to be the site of gravity sensing in roots because removal of the cap abolishes root curvature. Circumstantial evidence favors the columella cells as the gravisensory cells because amyloplasts (and often other cellular components) are polarized with respect to the gravity vector. However, there has been no functional confirmation of their role. To address this problem, we used laser ablation to remove defined cells in the cap of Arabidopsis primary roots and quantified the response of the roots to gravity using three parameters: time course of curvature, presentation time, and deviation from vertical growth. Ablation of the peripheral cap cells and tip cells did not alter root curvature. Ablation of the innermost columella cells caused the strongest inhibitory effect on root curvature without affecting growth rates. Many of these roots deviated significantly from vertical growth and had a presentation time 6-fold longer than the controls. Among the two inner columella stories, the central cells of story 2 contributed the most to root gravitropism. These cells also exhibited the largest amyloplast sedimentation velocities. Therefore, these results are consistent with the starch-statolith sedimentation hypothesis for gravity sensing.

Arabidopsis↗