Effect of artificial gravitation on the skeletal musculature of rats during space flights]

Comparative morphological and histochemical investigation of the rat skeletal muscles after a 18.5 days flight in the space ship "Cosmos-936" under conditions of weightlessness and artificial gravitation created by means of a centrifuge has demonstrated that weightlessness creates muscular atrophy, with fibres of different types affected in different muscles. The greatest changes take place in the soleus muscle. Metabolic disorders in the muscular tissue are manifested as an excessive deposition of glycogen and an increased content of phospholipids in it. The artificial gravitation favourably affects the state of the skeletal musculature preventing the development of metabolic disorders, though it does not completely avert the appearance of atrophic processes in it.     

Skeletal muscle adaptation in rats flown on Cosmos 1667

Seven male Wistar rats were subjected to 7 days of weightlessness on the Soviet biosatellite Cosmos 1667. Muscle histomorphometry and biochemical analyses were performed on the soleus (SOL) and extensor digitorum longus (EDL) of flight rats (group F) and compared with data from three groups of terrestrial controls: one subjected to conditions similar to group F in space except for the state of weightlessness (group S) and the others living free in a vivarium (V1, V2). Relative to group V2 (its age and weight-matched control group), group F showed a greater decrease of muscle mass in SOL (23%) than in EDL (11%). In SOL a decrease in the percentage of type I fibers was counterbalanced by a simultaneous increase in type IIa fibers. The cross-sectional area of type I fiber was reduced by 24%. No statistically significant difference in capillarization and enzymatic activities was observed between the groups. In EDL a reduction in type I fiber distribution and 3-hydroxyacyl-CoA-dehydrogenase activity (27%) occurred after the flight. The small histochemical and biochemical changes reported suggest the interest in studying muscular adaptation during a flight of longer duration.     

Compensatory-adaptive modifications of the C-cell apparatus of the thyroid in normal rats of various ages and in partial sympathectomy

Population of the thyroid C-cells, normal and at sympathectomy has been analyzed in 75 white male rats at the age of 1, 3 and 6 months by means of electron microscopical, morphometrical and radioimmunological methods. Partial sympathectomy has been performed using subcutaneous injection of guanethidine (15 mg/kg of body mass) during 14 days after birth. During the period from 1 up to 6 months of life in intact rats a decrease in C-cells functional activity is observed. Under conditions of sympathectomy in 30-day-old animals decreasing extrusion processes of the secretory material are observed. During successive periods of life (3 and 6 months) mechanisms of paracrinic evacuation of hormonal products enhance considerably, nuclear volume of the cells and number of C-cells in the field of vision increase. Their hyperplastic alterations in the sympathectomized thyroid gland are more pronounced in 3-month-old animals.     

The effect of space flight conditions on the rate of multiplication, morphology of cells, DNA and protein content in the ciliate Tetrahymena pyriformis

The conditions of a space flight and, in particular, the weightlessness promote an increased density of the ciliate culture, an enhanced reproduction rate, and an elevated ratio of dividing cells. The condition of weightlessness brings about some decrease in the bulk protein content of the cells determined by cytophotometry of Naphthol-yellow stained ciliates. The quantity of DNA in macronuclei was measured following the routine Feulgen procedure (its "cold" variant). The DNA content was found to remain unchanged. Some changes in the shape and size of the cells were noticed under flight conditions: ciliates that had developed in weightlessness appeared more spherical than control ones, due presumably to a decrease in the body length and to some extension in the body width. The conditions of space flight, including the weightlessness, induce changes in the physiological status of unicellular organisms. A decrease in the gravitation force may lead to a decrease in the energy expenditures for maintenance of the cell positional homeostasis.     

Experiments with cultures of mammalian cells aboard the biosatellite "Cosmos-782"

A considerable contribution to the investigation on biological importance of weightlessness was made by the experiments with animals in the artificial Earth satelites (AES) of "Cosmos" type. Cell cultures can serve as an ideal model to get a direct cell response to the effect of external factors. For the experiment in the AES "Cosmos-782", two thoroughly examined cell strains (L and 237) were chosen, which differed in a number of parameters (for example, duration of their mitotic cycles). Density of cell seeding and temperature of their cultivation in the laboratory experiment were calculated in such a way that the whole cycle of the culture development should take place under the conditions of weightlessness: the beginning of lag-phase--before launching and the stationary phase--after landing. The weightlessness was not shown to result in any genetical shifts revealed at chromosomal level. When cultivated after the flight, the cells do not change their mitotic cycle parameters, mitotic course and structural organization. The data obtained in the experiments with AES "Cosmos-368" and "Cosmos-782" (increase of mitotic index, some forms of mitotic pathology during the first terms of cultivation after the flight and enlargement of cellular nuclei) demonstrate the changes in the cell population which have formed under the conditions of weightlessness. Similar changes are observed while the cells propagate in the laboratory conditions. Indirect data on an earlier cell culture aging during the flight do not exclued the possibility that under weightlessness the rate of cell propagation could differ from that under gravitation.     

Morphological and biochemical examination of Cosmos 1887 rat heart tissue: Part I--Ultrastructure

Morphological changes were observed in the left ventricle of rat heart tissue from animals flown on the Cosmos 1887 biosatellite for 12.5 days. These tissues were compared to the synchronous and vivarium control hearts. While many normal myofibrils were observed, others exhibited ultrastructural alterations, i.e., damaged and irregular-shaped mitochondria and generalized myofibrillar edema. Analysis of variance (ANOVA) of the volume density data revealed a statistically significant increase in glycogen and a significant decrease in mitochondria compared to the synchronous and vivarium controls. Point counting indicated an increase in lipid and myeloid bodies and a decrease in microtubules, but these changes were not statistically significant. In addition, the flight animals exhibited some patchy loss of protofibrils (actin and myosin filaments) and some abnormal supercontracted myofibrils that were not seen in the controls. This study was undertaken to gain insight into the mechanistic aspects of cardiac changes in both animals and human beings as a consequence of space travel (1). Cardiac hypotrophy and fluid shifts have been observed after actual or simulated weightlessness and raise concerns about the functioning of the heart and circulatory system during and after travel in space (2-4).     

Biochemical changes in rat liver after 18.5 days of spaceflight

The effect of "weightlessness" on liver metabolism was examined using tissue from rats flown in earth orbit for 18.5 days aboard the Soviet Cosmos 936 biosatellite. Changes in the activities of certain carbohydrate and lipid enzymes were noted. Of the 28 hepatic enzyme activities assayed, two, palmitoyl-CoA desaturase and lactate dehydrogenase, increased, whereas five, glycogen phosphorylase, 6-phosphogluconate dehydrogenase, both acyltransferases which act on alpha-glycerolphosphate and diglycerides, and aconitate hydratase decreased. The remaining enzyme activities measured were unchanged. In addition, increased levels of liver glycogen and palmitoleate were noted which probably resulted from the lowered glycogen phosphorylase and increased palmitoyl-CoA desaturase activities, respectively, in those animals that experienced weightlessness. These changes caused by weightlessness were transient since all of the aforementioned alterations returned to normal values when measured in the livers of other rats which had flown in the biosatellite 25 days after recovery.     

Quantitative changes in the frequency and distribution of the C-cell population in the rat thyroid gland with age

Summary The development of calcitonin cells (C-cells) was investigated in rat thyroid glands from birth to 120 days, using an immunoperoxidase technique and a point-counting method. The proportion of C-cells to follicular cells was 4.5% on the day of birth and increased progressively to 10.4% by 120 days. The highest density of C-cells was noted in the mid-region of the lobes along a longitudinal axis. The caudal and cephalic regions of the lobes contained smaller numbers of C-cells. The C-cells tended to be more numerous in the posterior aspects of the lobes. Although the numbers of C-cells in 120-day-old animals were markedly increased as compared to animals at the time of birth, the cell distributions within the glands were similar at all ages.     

Influence of gravitational overload and hypokinesia on the structure of the vascular bed of the stomach]

Under study were effects of gravitation stresses, total hypokinesia and their combinations on blood vessels of the stomach. The work was carried out in 130 rabbits, 16 of them being used to study the normal structure of the gastric vascular bed. The vascular bed was injected with the Gerota's mass followed by clearing, making histological preparations and roentgenography. The investigation has revealed both quantitative and qualitative changes in the structure of the gastric blood vessels. The maximum endurable stress of the ventro-dorsal direction causes morphological shifts less pronounced than stresses of longitudinal directions. With prolonged terms of hypokinesia (1-12 weeks) morphological changes became more pronounced in all the layers of the stomach. A combination of successive gravitation stresses and hypokinesia during 4-12 weeks aggravated morphological changes which occurred after exposure to isolated above factors. The animals subjected to maximum endurable stresses before and after 4-week hypokinesia developed vascular changes more typical for the effects of gravitation. The pretraining of animals did not give a pronounced positive effect on the changes of the angioarchitectonics of all the gastric layers after a repeated stress following 4-week hypokinesia.     

Parameters of the innate and adaptive immunity in cosmonauts after long-term space flight on board the international space station

The results of the study of the innate and adaptive immunity indicators in 12 cosmonauts who took part in long-term (128–215 days) expeditions on board the International Space Station (ISS) are presented. It was shown that the space flight may lead to deviations in the human immune system. A decrease in the functional activity of phagocytes, NK and T-cells, as well as in the ability of immunocompetent cells to synthesize cytokines were observed. Significant individual changes were observed in the immune system’s response to a long-term space flight, which indicated an individual’s predisposition to the development of immune reactivity disorders under varying gravitation conditions.     

The water-salt balance and renal function in space flights and in model experiments

Study of a condition of mineral and water-electrolyte metabolism, function of kidneys, and their hormonal regulation during model experiments (hypokinesia, bed rest, immersion etc.), and also in space flights and in readaptation period, has shown a major role of water-electrolyte homeostasis during general adaptation of humans and animals to new conditions of life and to conditions of weightlessness in particular. The change in regulation of volumes of fluid milieu in an initial period of weightlessness was shown to be the consequence of redistribution of blood and hemodynamics of the shifts resulting in change of production of volume-regulation hormones, formation of negative water balance, and redistribution of fluid in the organism among various fluid compartments. At later stages of flight or long-term hypokinesia, a change of water-electrolyte homeostasis occurs with a decrease in the kidneys excretion of sodium, and diuresis, but with an increased excretion of calcium and production of ADH and RAAS hormones. Following returning to earth gravitation, the majority of astronauts have adaptive reactions, compensating for the loss extracellular fluid and mineral substances and formation of "earth" water-electrolyte homeostasis. For estimation of water-electrolyte homeostasis and the functions of kidneys in astronauts, various functional loading tests have been developed. The developed system of preventive maintenance is successfully used for abolition of adverse changes at various stages of space flight and in readaptation period.     

Effects of ethanol on the ultrastructure of the hamster thyroid C-cell

The morphology of the thyroid C-cells in golden hamsters after short- and long-term treatment with ethanol was studied. Immunohistochemistry was applied to examine the distribution of the C-cells in the thyroid gland. In the short-term experimental animals, the Golgi complexes and the granular endoplasmic reticulum were well developed and the number of the secretory granules was decreased as compared with those of the control animals. These findings suggest that the cellular activity of the thyroid C-cell is stimulated after short-term treatment with ethanol. The morphology of the thyroid C-cells of the long-term experimental animals was similar to that of the controls. It is conceivable that long-term treatment with ethanol does not affect the function of the C-cell.     

Structural and metabolic characteristics of human soleus fibers after long duration spaceflight

The fiber size decline, alterations in fiber metabolic potential and increase of connective tissue component were shown in human m. vastus lateralis after short and long-duration space flights and in m.soleus and m.vastus lateralis after 120 day head down tilt bed rest. It is known from rat and monkey studies that the exposure to weightlessness leads to the most pronounced changes in postural muscles, e.g. m.soleus. It was shown that 17 day space flight induced significant decrease of fiber cross-sectional area and slow-to-fast fiber type transformation in human soleus. But in the cited work the fiber population under study was limited like in most single fiber technique analyses. The present study was purposed to investigate the structural and metabolic properties of soleus muscle in Russian cosmonauts exposed to 129-day space flight on board of the International Space Station.     

Vestibular function and space motion sickness

The vestibular system plays an important role in intersensory interactions and gravitation is a natural stimulus for its receptors. Weightlessness alters the input signals of the otoliths and their effect on the pattern and dynamics of changes in the vestibular function (VF), which is accompanied by development of space adaptation syndrome (SAS) and space motion sickness (SMS). These changes occur both during the spaceflight (SF) and after returning to Earth, but the mechanisms of their development are still poorly understood and require special studies. In total, 47 Russian cosmonauts (crewmembers of long-term International Space Station (ISS) missions) have participated in the studies into VF before and after SF and nine of them, in onboard studies during SF (129–215 days) as a part of the Virtual space experiment (stage 1). Electro- and video-oculography are used to record spontaneous eye movements (SpEM), static vestibular–ocular responses during head tilts to the right or left shoulder (static otolith–cervical–ocular reflex, OCOR), and dynamic vestibular-ocular response during the head rotation around the longitudinal axis of the body. The examination is accompanied by personal and questionnaire survey on subjective responses and complaints of cosmonauts about SAS and SMS. Significant changes in SpEM (drifts of eyes, spontaneous and gaze-evoked nystagmus, and arbitrary saccades) and a decrease in OCOR (statistically significant decrease in the amplitude of ocular counter-rolling in response to head tilts up to its absence or inversion, an atypical OCOR) are observed during SF. An atypical OCOR is observed at the beginning of adaptation to weightlessness in seven of the nine cosmonauts (the first one to two weeks of SF) and repeatedly throughout the flight in all cosmonauts regardless of whether it is their first flight or not. Atypical vestibular responses after SF, similar to the responses during SF, are observed in several cosmonauts by day 9 after flight. It has been shown that atypical OCOR variants are more frequently observed in the subjects lacking any previous space experience, as well as a more pronounced decrease in this response with a concurrent increase in the response of the semicircular canals. It is also demonstrated that repeated SFs lead to a considerable shortening in the after-flight readaptation to terrestrial conditions and a considerable decrease in the degree of vestibular disorders. In the initial period of SF, the changes in VF are correlated with the complaints and manifestations of SAS and SMS; however, the complaints and the corresponding symptoms are unobservable during the further flight despite significant changes in the VF state. The patterns of the VF disorders associated with the impact of weightlessness and observed during and after SF are very similar, allowing these disorders to be regarded as SAS and SMS of different severities (intensities).     

Histometry of normal thyroid glands in neonatal and adult rats

The present histometric study is on thyroid glands of Wistar rats ranging in age from 0 to 120 days. The mean volume of one lobe of the thyroid in 4-month-old animals was some 22-, 10-, 5-, and 3-fold greater, respectively, than the volumes in the newborn, 5-, 10-, and 30-day-old rats. At 4 months of age the mean length of the lobe was 3 times greater than at birth. The volumetric fractions (Vv) of the different histological components (follicular cells, C-cells, colloid, and interstitial tissue) changed considerably in the course of development. The Vv of follicular cells diminished from 61.4% at birth to 37.2% at 4 months. C-cells increased from 2.9% in the newborn to 4% at 15 days, with no further significant change at 4 months. Colloid and stroma together represented 35.7% at birth, increasing to 58.5% at 120 days. In the course of the first 4 months of life, the absolute volumes occupied by follicular cells, C-cells, colloid, and stroma increased 13.25, 30.75, 38.6, and 33.7 times, respectively; these changes reflect the variations that occur in the volume of the gland and the Vv throughout postnatal development.     

Complex estimation of thyroid gland state in tundra voles, inhabiting the natural radioactive polluted sites

Behow is presented the complex state of thyroid gland of tundra voles living in high level natural radioactivity conditions (Ukhta region of Komi Republic) by morphological and functional criteria. High sensitivity of thyroid gland under natural chronic irradiation of whole organism is noticed. The essential changes in the morphological structure and in the hormone level of the thyroid gland are found in animals, which live for a long time in the conditions of the radioactive pollution. It is determined, that the inside population processes influence on the structure and on the funcition of the thyroid gland.     

Lipid peroxidation rate and the antioxidant protection system during the readaptation period after long-term space flights on board the International Space Station

The content of lipid peroxidation (LPO) products (diene conjugates (DC), malondialdehyde (MDA), Schiff bases (SB), and tocopherol (TP, a main lipid antioxidant) were measured in blood serum of 17 astronauts taking part in long-term (125–217 days) missions on board the International Space Station (ISS) during the preflight period, on the day of the landing, and on the 7th and 14th days after landing (the rehabilitation period, RP). A decrease in the DC and MDA levels against a background of an increase in TP has been found in a group of eight astronauts after landing on board the Space Shuttle spacecraft and a group of eight astronauts after a space flight on board the Soyuz TM in the course of RP. The changes in measured indices were more pronounced in the group of astronauts after the space flight on board the Space Shuttle spacecraft. Inhibition of LPO during RP was regarded as an adequate response to readaptation stress to the conditions on earth. The possible mechanisms of differences in the efficiency of LPO inhibition between groups are discussed: the changes in the biomembrane phase state under the conditions of deceleration load during disorbiting and the stressful reaction to landing on board different spacecrafts.     

Postnatal variations in the number and size of C-cells in the rat thyroid gland

The heterogeneous distribution of thyroid C-cells has until now hindered an objective evaluation of changes caused by age or experimental stimuli. To overcome this, a rigorous methodology has been designed to detect variations in shape, size, and number of C-cells throughout development. Using this methodology, we have demonstrated that C-cells do not significantly alter their shape with age. However, their volume increases gradually from 472 m³ in newborn rats to 1653 m³ in 120-day-old animals. Over the same time period, the mean number of C-cells within the thyroid gland increased 9-fold (from 1.6x10⁴ to 1.5x10⁵), and the number of C-cells per unit area decreased (from 6.15x10⁴/mm³ to 2.6x10⁴/mm³). We conclude that there are marked variations in size, total number, and number of C-cells per unit area in the rat thyroid gland after birth.     

Changes in myocardial contractility and contractile proteins after four weeks of simulated [correction of simulate] weightlessness in rats

The interaction between the gravitational field, the position of the body, and the functional characteristics of the blood vessels determines the distribution of intravascular volume. In turn, this distribution determines cardiac pump function. One of the most profound circulatory changes that occurs in man during exposure to weightlessness is a cephalad redistribution of fluid caused by the lack of hydrostatic pressure in this microgravitative environment. The cephalad redistribution of fluid results in a loss of blood volume and then induces a decrease in preload. Recently, a decrease in sensitivity of arteriole to catecholamine has reported in rats of simulated weightlessness. This change in arteriole may reduce afterload. As a result, cardiovascular system may be shifted to a hypokinetic state during weightlessness condition for long-term. Echocardiographic data from astronauts during space flight showed an increase in heart rate, a 12 % decrease in stroke volume, and a 16 % decrease in left end diastolic volume. Electron-microscopic studies have shown changes in cardiac morphology in rats after exposure to microgravity for 7-12.5 days. After the COSMOS 2044 flight for 14 days, the light-microscopic studies have shown an atrophy of papillary muscles in rats left cardiac ventricle. It is not clear whether the function of atrophic myocardium is impaired. The data in three aspects as mentioned above suggest that weightlessness or simulated weightlessness may decrease the myocardial function. However, definite changes in cardiac performance have been hard to prove due to many limits. This studies were to answer two questions: Is the myocardial contractility depressed in rats subjected to simulated weightlessness for four weeks? What are the underlying mechanisms of the changing contractility?     

Behavioral and functional vestibular disorders after space flight: I. Mammals

The review presents data on functional disorders in mammals caused by changes in the vestibular system after space flight. These data show that the mammalian vestibular system responds to weightlessness dissimilarly at different ontogenetic stages. During the embryonic period, orbital space flight conditions have a little effect on the developing vestibular system and even promote normal fetal development. During the early postnatal period, when optimal sensorymotor tactics arise, long-term exposure to space flight conditions leads to the development of novel, “extraterrestrial”, sensory-motor programs that may fixate in CNS for life. In adult individuals, substantial vestibular changes and disorders may occur immediately after landing depending on the weightlessness duration. An adult organism has to solve two concurrent and mutually conflicting problems: to adapt to weightlessness and not to adapt to it in order to facilitate readaptation after return. Thus, individuals have to counteract weightlessness to retain a maximum of their pre-flight health status. The means of such a counteraction have to be adjusted according to the weightlessness duration. It is noteworthy, however, that not all functional changes occurring in adult individuals under weightlessness can be adequately accounted for. Some of them can assume a chronic or even pathological character. The review raises for the first time the question of necessity to include into the scope of studies the effect of weightlessness on a senile (senescent) organism and its vestibular system. We believe that development of space gerontology as a special branch of space biology and medicine is undoubtedly of interest and may become practically important in the future in view of the ever-growing age of space explorers.