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Biomedical subjects

R Tixador

Publications and source records attributed to R Tixador.

At least 19 recordsLinked to original sources

Skeletogenesis in sea urchin larvae under modified gravity conditions.

From many points of view, skeletogenesis in sea urchins has been well described. Based on this scientific background and considering practical aspects of sea urchin development (i.e. availability of material, size of larvae, etc.), we wanted to know whether orderly skeletogenesis requires the presence of gravity. The objective has been approached by three experiments successfully performed under genuine microgravity conditions (in the STS-65 IML-2 mission of 1994; in the Photon-10 IBIS mission of 1995 and in the STS-76 S/MM-03 mission of 1996). Larvae of the sea urchin Sphaerechinus granularis were allowed to develop in microgravity conditions for several days from blastula stage onwards (onset of skeletogenesis). At the end of the missions, the recovered skeletal structures were studied with respect to their mineral composition, architecture and size. Live larvae were also recovered for post-flight culture. The results obtained clearly show that the process of mineralisation is independent of gravity: that is, the skeletogenic cells differentiate correctly in microgravity. However, abnormal skeleton architectures were encountered, particularly in the IML-2 mission, indicating that the process of positioning of the skeletogenic cells may be affected, directly or indirectly, by environmental factors, including gravity. Larvae exposed to microgravity from blastula to prism/early pluteus stage for about 2 weeks (IBIS mission), developed on the ground over the next 2 months into normal metamorphosing individuals.

Animals↗

The sea urchin larva, a suitable model for biomineralisation studies in space (IML-2 ESA Biorack experiment '24-F urchin').

By the ESA Biorack 'F-24 urchin' experiment of the IML-2 mission, for the first time the biomineralisation process in developing sea urchin larvae could be studied under real microgravity conditions. The main objectives were to determine whether in microgravity the process of skeleton formation does occur correctly compared to normal gravity conditions and whether larvae with differentiated skeletons do 'de-mineralise'. These objectives have been essentially achieved. Postflight studies on the recovered 'sub-normal' skeletons focused on qualitative, statistical and quantitative aspects. Clear evidence is obtained that the basic biomineralisation process does actually occur normally in microgravity. No significant differences are observed between flight and ground samples. The sub-normal skeleton architectures indicate, however, that the process of positioning of the skeletogenic cells (determining primarily shape and size of the skeleton) is particularly sensitive to modifications of environmental factors, potentially including gravity. The anatomical heterogeneity of the recovered skeletons, interpreted as long term effect of an accidental thermal shock during artificial egg fertilisation (break of climatisation at LSSF), masks possible effects of microgravity. No pronounced demineralisation appears to occur in microgravity; the magnesium component of the skeleton seems yet less stable than the calcium. On the basis of these results, a continuation of biomineralisation studies in space, with the sea urchin larva as model system, appears well justified and desirable.

Animals↗

Influence of the environment in space on the biochemical characteristics of human low density lipoproteins.

The purpose of this experiment was to study the efficiency of protective substances on the effects of cosmic radiation in space on low density lipoproteins. This environment induced modifications in LDL consisting of an increase of lipid peroxidation markers (hydroperoxides, thiobarbituric acid reactive substances). In contrast, apo B was not affected by cosmic radiation as shown by the stability of the trinitrobenzenesulfonic acid reactivity and the tryptophan content. Furthermore, oxidation of LDL was partially inhibited by the addition of cysteamine or/and probucol before the spaceflight experiment. The hydroperoxide formation was almost completely inhibited by cysteamine. It was concluded that antioxidants can exert a protective effect against peroxidative stress induced by the space environment.

Cosmic Radiation↗

Growth and division of Escherichia coli under microgravity conditions.

The growth rate in glucose minimal medium and time of entry into the stationary phase in pepton cultures were determined during the STS 42 mission of the space shuttle Discovery. Cells were cultured in plastic bags and growth was stopped at six different time points by lowering the temperature to 5 degrees C, and at a single time point, by formaldehyde fixation. Based on cell number determination, the doubling time calculated for the flight samples of glucose cells was shorter (46 min) than for the ground samples (59 min). However, a larger cell size expected for more rapidly growing cells was not observed by volume measurements with the electronic particle counter, nor by electron microscopic measurement of cell dimensions. Only for cells fixed in flight was a larger cell length and percentage of constricted cells found. An optical density increase in the peptone cultures showed an earlier entry into the stationary phase in flight samples, but this could not be confirmed by viability counts. The single sample with cells fixed in flight showed properties indicative of growth stimulation. However, taking all observations together, we conclude that microgravity has no effect on the growth rate of exponentially growing Escherichia coli cells.

Cell Division↗

Behavior of bacteria and antibiotics under space conditions.

We have previously reported an increase of the "resistance" to antibiotics of bacteria during space missions. In the present experiment, we studied the growth of Escherichia coli cultured in vitro in space in the presence of dihydrostreptomycin: tritiated and nontritiated. This experiment was carried out during the STS 42 mission aboard the U.S. Space Shuttle Discovery (IML-1 program). Cells were cultured in plastic bags and growth was stopped at six different time points by lowering the temperature to 5 degrees C. Several methods were used: viable cell counting by Colony Forming Units; total cell number by optical densitometry; electron microscopy; radioactivity measurements. The investigations show no difference between flight and ground experiments for the cultures without antibiotic. The growth rate with antibiotic was accelerated in flight, the growth yield was not changed, and there were no differences in the ultrastructures. The results suggest some changes in antibiotic binding in space. We did not observe any differences between the cultures developed in flight in the 1-g centrifuge and the cultures placed in the static rack in microgravity.

Colony Count, Microbial↗

Influence of low-temperature storage and glucose starvation on growth recovery in Escherichia coli relA and relA+ strains.

To study the influence of microgravity on bacterial growth behavior during a space mission, the special experimental conditions and the hardware environment necessitate storage of cells at low temperature, and permit a relatively short experimental period. Before this experimental period, cells have to recover their condition of steady-state growth, because it is only in this condition that the growth behavior of the flight and ground populations can be adequately compared. To meet these requirements and to obtain cells which recover rapidly their steady-state growth, we analyzed the size and shape of Escherichia coli cells during storage at 4 degrees C, with and without previous glucose starvation of the cells. It appeared that cells stored at low temperature in the presence of glucose continued to increase in average mass and assumed ovoid shapes. In addition, upon restoration of maximal growth rate at 37 degrees C, they continued to increase in size and showed a transient overshoot of their final steady-state value, which was reached after about 5 h. Cells previously starved for glucose, however, maintained their average size and rod-shape during low-temperature storage. Recovery of the starved cells was most rapid in the relA+ strain which, contrary to the isogenic relA strain, showed no overshoot and reached its final steady-state size within 2 h.

Cold Temperature↗

Effects of angular speed in responses of Paramecium tetraurelia to hypergravity.

The paper shows the results of investigations carried out in a single cell organism. Paramecium tetraurelia exposed to different gravitational levels. Hypergravity resulted in a decrease in cell growth rate. The responses depend on g level and angular speed of the centrifuge; furthermore they depend also on small short fluctuations in g levels, delta g, due to the swimming of the cells inside the culture tubes. Delta g depends on angular speed and size of the holding device. The inhibitory effect of hypergravity, for the same angular speed, increases with respect of the diameter of the culture tubes.

Animals↗

Effects of microgravity and hypergravity on the cell: investigations on Paramecium tetraurelia.

Previous space CYTOS experiments have shown that space flights resulted in an increase in growth of Paramecia cultures. Microgravity is the major factor responsible of this response: indeed the stimulatory effect disappeared in inflight cultures placed on a 1 g centrifuge aboard the Spacelab. On the other hand, exposure to different levels of hypergravity on Earth resulted in an opposite response, i.e. to a reduced cell growth rate. A possible mechanism of microgravity on paramecia is discussed.

Animals↗

Influence on cell proliferation of background radiation or exposure to very low, chronic gamma radiation.

Investigations carried out on the protozoan Paramecium tetraurelia and the cyanobacteria Synechococcus lividus, which were shielded against background radiation or exposed to very low doses of gamma radiation, demonstrated that radiation can stimulate the proliferation of these two single-cell organisms. Radiation hormesis depends on internal factors (age of starting cells) and external factors (lighting conditions). The stimulatory effect occurred only in a limited range of doses and disappeared for dose rates higher than 50 mGy/y.

Animals↗

Antibiotic activity in space.

Environmental factors in space exert an influence on the behaviour of bacteria, particularly on their sensitivity to antibiotics. Thus, G. Taylor and S. Zaloguev observed that bacterial samples collected on the crew during flight in the Apollo-Soyouz Test Project Mission presented higher antibiotic resistance than controls. This paper presents the results of two experiments performed in 1982 and 1985 (Cytos 2 during the French-Soviet Mission and "Antibio" in the Biorack programme of the European Space Agency). The results show an increase of antibiotic resistance in bacteria growth in flight and a modification in the structure of the cell wall. All these modifications are transitory. Two hypotheses are put forward to explain the phenomenon.

Anti-Bacterial Agents↗

Respective role of microgravity and cosmic rays on Paramecium tetraurelia cultured aboard Salyut 6.

Paramecium tetraurelia cultured aboard Salyut 6 have shown in increase in cell growth rate, cell volume, water content and changes in electrolyte content. Additional experiments, carried out in balloon flight and on earth, showed that the stimulating effect observed on cell proliferation is related to exposure to cosmic rays. Other changes seem to be due to a direct effect of microgravity on cell. Mechanism of gravity action on cell is discussed.

Animals↗

Preliminary results of Cytos 2 experiment.

Cytos 2 experiment, carried out during the French-Soviet manned flight (July 1982), has studied the antibiotics sensitivity of bacteria cultivated in vitro during the orbital flight. The results show an increase of the antibiotics resistance and a larger thickness of the cellular envelope for the inflight cells. The increase of antibiotics resistance can be related to a stimulating effect of space on the cell growth rate or to changes of the cellular envelope structure.

Anti-Bacterial Agents↗

Study of minimal inhibitory concentration of antibiotics on bacteria cultivated in vitro in space (Cytos 2 experiment).

The aim of the Cytos 2 experiment, carried out during the French-Soviet manned flight in July 1982, was to study the bacteria's sensitivity to antibiotics cultivated in vitro during the orbital flight, using the bacterial method of minimal inhibitory concentration (MIC). Two species of bacteria were tested with various antibiotics: Staphylococcus aureus with Oxacillin, Chloramphenicol and Erythromycin; Escherichia coli with Colistin and Kanamycin. The results show an increase in resistance to antibiotics particularly strong in E. coli and weaker in Staphylococcus aureus. Considering these results, we think that there might be a relationship between the increase in resistance to antibiotics and a stimulating effect on growth rate by the factors of environmental space.

Anti-Bacterial Agents↗

Effects of gravity and cosmic rays on cell proliferation kinetics in Paramecium tetraurelia.

Space flights resulted in a stimulating effect on kinetics of proliferation in Paramecium tetraurelia. Additional experiments were performed in order to determine the origin of this phenomena. Paramecia were cultivated in balloon flights or in a slow clinostat, or were exposed to different levels of hypergravity. The results suggest that changes in cell proliferation rate are related to cosmic rays and to a direct effect of microgravity.

Animals↗

[Biological effects of weightlessness at the cellular level. Comparative study of cultures of Paramecia aboard the orbital station Salyut-6 and a stratospheric balloon].

In order to distinguish the effects of cosmic rays from those of weightlessness at the cellular level, we performed experiments aboard stratospheric balloon, where gravity is equal to 1 g and cosmic radiation roughly equal to that aboard Salyut-6. The results suggest that the stimulation of cell proliferation is probably due to cosmic rays, metabolic changes being related to microgravity.

Calcium↗