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Biomedical subjects

A Cogoli

Publications and source records attributed to A Cogoli.

At least 55 records · Page 3Linked to original sources

Space flight and the immune system.

Depression of lymphocyte response to mitogens in cosmonauts after space flight was reported for the first time in the early 1970s by Soviet immunologists. Today we know that depression of lymphocyte function affects at least 50% of space crew members. Investigations on the ground on subjects undergoing physical and psychological stress indicate that stress is a major factor in immune depression of astronauts. This is despite the fact that weightlessness per se has a strong inhibitory effect on lymphocyte activation in vitro. Although the changes observed never harmed the health of astronauts, immunological changes must be seriously investigated and understood in view of long-duration flight on space stations in an Earth orbit, to other planets such as Mars and to the Moon.

Animals↗

Cultivation of hamster kidney cells in a dynamic cell culture system in space (Spacelab IML-1 Mission).

Cell proliferation, tissue plasminogen activator (t-PA) production and metabolic changes of Hamster Kidney cells (HaK) grown on microcarriers in an automatic Dynamic Cell Culture System (DCCS) were determined on the first International Microgravity Mission (IML-1) Spacelab (22-30 January 1992). The DCCS was designed for two cell culture chambers (volume: 200 microliters each), one operating as a hatch system, the other as a perfusion system. Medium exchange was achieved with an osmotic pump (flow rate 1 microliter h-1). Two major items were investigated: the biological performance of the DCCS in space and the effect of microgravity on HaK cells. The results obtained demonstrated that (1) the DCCS can be used for biological experiments on long term Spacelab missions. In fact, higher cell densities and higher concentration of glucose but lower concentration of lactate in the perfusion chambers than in the batch chambers were measured. The concentration of t-PA, glutamine and ammonia was similar in all chambers. (2) Microgravity had no effect on cell growth and metabolism of HaK cells.

Ammonia↗

The effect of space flight on human cellular immunity.

T-lymphocyte responsiveness to mitogens is depressed by an average of 56% in humans (129 subjects) tested during and after space flight. Although there is not yet conclusive evidence of a clinical significance of the test, it is clear that factors of space flight like stress, closed environment and cosmic radiation may affect immune responsiveness. The data obtained from space crews may be compared to the depression seen in subjects undergoing heavy physical stress of head down tilt bedrest. Recently, delayed hypersensitivity [correction of hypersensivity] response was tested on crews of the US space shuttle and of the orbital station MIR by means of a commercially available "skin test". Again, the response was lower in 14 of the 15 subjects tested. In two cases, a strong in flight depression could be related to heavy physical and psychological stress experienced in flight. The data available today are not sufficient to draw conclusions on the hazard of infectious diseases during and after space flight. Although the changes observed never harmed the health of astronauts, immunological changes must be seriously investigated and understood in view of long-duration flights on space stations in an Earth orbit and to other planets like Mars and the Moon.

Adult↗

Activation of microcarrier-attached lymphocytes in microgravity.

A technology has been developed to achieve optimal attachment of adhesion-independent lymphocytes to microcarrier beads. The activation of T-lymphocytes by concanavalin A was tested under microgravity conditions in an experiment carried out in space during the first Spacelab Life Science Mission. Activation, measured as the synthesis of deoxyribonucleic acid (DNA) and the production of interferon-gamma, more than doubled in attached lymphocytes in microgravity. The depression of the activation discovered in previous space experiments is due to an impairment not of the lymphocyte but of the macrophage function. The system described here may be useful for radiobiological investigations on the effect of high-energy particles and for testing the efficiency of the immune system in humans during the long-duration space flight planned in the future. The biotechnological significance of the increased lymphokine production in space remains to be assessed.

Biotechnology↗

Reduced lymphocyte activation in space: role of cell-substratum interactions.

We investigated the effect of substratum adhesiveness on stimulated lymphocyte blastogenesis by reducing and blocking cell adhesion with poly (2-hydroxyethyl methacrylate) (poly-HEMA) in a simple on-ground system. Cells grown on medium-thick and thick poly-HEMA films were rounded in shape and displayed no signs of spreading. By contrast, on tissue culture plastic and very thin poly-HEMA films, they showed clear signs of spreading. The mitogenic response of lymphocytes grown on thick poly-HEMA films was reduced by up to 68% of the control (tissue culture plastic). Interferon-gamma production was near zero when the cells were grown on the least adhesive substratum. On uncoated plastic, activated lymphocytes subjected to high gravity (20g) exhibited an increased proliferation rate (40%) compared with 1g. By contrast, on poly-HEMA, high gravity did not improve lymphocyte responsiveness. These results show that activated lymphocytes need to anchor and spread prior to achieving an optimal proliferation response. We conclude that decreased lymphocyte adhesion could contribute to the depressed in vitro lymphocyte responsiveness found in the microgravity conditions of space flight.

Cell Adhesion↗

Effect of head-down tilt bedrest (10 days) on lymphocyte reactivity.

Immunological responses of six healthy males to 10 days of head-down tilt bedrest (HDT) were assessed. Lymphocyte responsiveness was severely reduced immediately before, during, and immediately after the HDT, even though the lymphocyte numbers did not change. By contrast, delayed-type hypersensitivity was not affected. No dramatic changes were found in WBC counts and lymphocyte subpopulations, with the only exception of natural killer (NK) cells which transiently decreased immediately after HDT. Plasma cortisol levels were elevated above normal immediately before and during the HDT. The data suggest that the mitogenic response of lymphocytes was affected by psychological and fluid shift stress. These results are compared with data obtained during and after spaceflight. We conclude that the stress of HDT induces changes in immunological responsiveness that are strikingly similar to those arising from the stress of spaceflight.

Adult↗

Changes observed in lymphocyte behavior during gravitational unloading.

The effect of microgravity has been extensively studied on human lymphocytes in several space missions. A clear distinction must be made between two kinds of experiments: (i) with cells purified from the peripheral blood of test subjects before flight and then exposed inflight to mitogens and other activators (these are called in vitro experiments), and (ii) with lymphocytes from crewmembers of space missions exposed to mitogens prior to and after flight (ex vivo experiments). The first approach can be considered as basic research in cell biology in space; the second contributes to identifying the effects of the stress of spaceflight on the immune response of astronauts. The results from in vitro experiments have clearly shown that lymphocyte activation is nearly totally depressed in microgravity. This activation depression is confirmed by investigations on Earth in the fast rotating clinostat. Conversely, activation is increased when lymphocytes are cultured at 10 g in a centrifuge. In microgravity cell adhesion may be reduced, thus partly accounting for the decreased cell activation. The results of the experiments conducted at 10 g are due to a simultaneous activation of T- and B-lymphocytes by concanavalin A. The reduced activation observed in lymphocytes from crewmembers of space missions can be ascribed to both the physical and psychological stress of spaceflight. This observation was confirmed by investigations on subjects undergoing stress on Earth.

B-Lymphocytes↗

A ground-based model to study the effects of weightlessness on lymphocytes.

The mitogenic response of human lymphocytes was found to be markedly reduced in weightlessness conditions as compared to normal gravity. One possible explanation is that due to the non-existent sedimentation in space the lymphocytes could not adhere and spread on a substratum. Thus, we investigated the effect of substratum adhesiveness on lymphocyte responsiveness by reducing and blocking cell adhesion with poly-HEMA in a simple on-ground system. Lymphocyte adhesiveness was assessed by measuring the proportion of non-adhesive, slightly, and strongly adhesive 51Cr-radiolabelled cells on uncoated and poly-HEMA coated plastic. The amount of cell spreading on surfaces with varying adhesiveness was determined by measuring the area of cells. Cells grown on medium and thick poly-HEMA films were rounded in shape. By contrast, on tissue culture plastic, they showed clear signs of spreading. The mitogenic response of lymphocytes grown on thick poly-HEMA films was reduced by up to 68% of the control (tissue culture plastic). Interferon-gamma production was virtually nil when the cells were grown on the least adhesive substratum. These results show that activated lymphocytes need to anchor and spread prior to achieving an optimal proliferation response. We conclude that decreased lymphocyte adhesion could contribute to the depressed in vitro lymphocyte responsiveness found in the microgravity conditions of space flight.

Cell Adhesion↗

Effect of a herbal yeast food supplement and long-distance running on immunological parameters.

The effect of a food supplement on immunological parameters of 16 long-distance runners was tested in a randomized, double-blind and placebo-controlled trial. The supplement comprised plasmolysed herbal yeast, malt, honey, and orange juice. No statistically significant differences between the two groups regarding the following variables were detected at three sessions at rest and immediately after a 21 km run: total and differential white blood cell counts, numbers of B- and T-cells and T-subpopulations, concanavalin-A-induced lymphocyte proliferation, serum levels of immunoglobulins, neopterin, IL-2 receptors, beta 2-microglobulin, complement factor b, c4 and c3c, and c1-inactivator. These findings suggest that the effects of the tested food supplement on these parameters are negligible with respect to improvements in the immunological status of long-distance runners. The changes observed immediately after the run had a transient character. In both groups, however, low lymphocyte counts, IgG subclass 2 levels and c1-inactivator levels were noted at rest, which indicate that the immune status of endurance athletes may be affected by training.

Adult↗

[Effect of dimethyladipimidate and dimethylsuberimidate on cell-cell adhesion in rat fibroblasts].

In a study performed to identify the molecular mechanisms which regulate cell to cell adhesion and contact inhibition in neoplastic and syngeneic normal cells of the rat we have observed that the adhesive capacity depends on the reagents used, either EDTA or trypsin, to release the cells from monolayer. Taking profit of this last property and of the possibility of blocking free -NH2 groups on membrane proteins with specific cross-linking reagents "in vitro", we have studied in this work the behaviour of the proteins of the cell coat involved in cell to cell adhesion of rat fibroblasts FG/2. The cross-linking reagents used were dimethyladipimidate (DMA) and dimethylsuberimidate (DMS). The cells were exposed to the reagents at 0 degrees C for 30'. Cell to cell adhesion was measured by determining the percentage of single cells labeled with 3H-leucine, adhering to a confluent monolayer at different incubation times. The inhibitory effect on cell to cell adhesion brought about by cross-linking reagents indicates that a) EDTA-released cells are more sensitive to both imides than those released with trypsin, b) DMA is more effective on trypsin-released cells and c) DMS is more effective on EDTA-released cells. Therefore, we conclude that the inhibition of adhesion by reaction with the two cross-linking reagents is more likely due to a stiffening of the molecules of the cell coat involved in the adhesion, rather than to the modification of -NH2 residues which should specifically participate to adhesive process.

Animals↗

Bioprocessing under microgravity--an introduction.

The main biological processes presently considered for applications in space laboratories are: (i) bioseparation, (ii) cell cultivation and (iii) cell electrofusion. All three technologies were discussed by experts in the field at the 1988-COSPAR Meeting in Helsinki.

Biotechnology↗

Mammalian cell cultivation in space.

Equipment used in space for the cultivation of mammalian cells does not meet the usual standard of earth bound bioreactors. Thus, the development of a space worthy bioreactor is mandatory for two reasons: First, to investigate the effect on single cells of the space environment in general and microgravity conditions in particular, and second, to provide researchers on long term missions and the Space Station with cell material. However, expertise for this venture is not at hand. A small and simple device for animal cell culture experiments aboard Spacelab (Dynamic Cell Culture System; DCCS) was developed. It provides 2 cell culture chambers, one is operated as a batch system, the other one as a perfusion system. The cell chambers have a volume of 200 microliters. Medium exchange is achieved with an automatic osmotic pump. The system is neither mechanically stirred nor equipped with sensors. Oxygen for cell growth is provided by a gas chamber that is adjacent to the cell chambers. The oxygen gradient produced by the growing cells serves to maintain the oxygen influx by diffusion. Hamster kidney cells growing on microcarriers were used to test the biological performance of the DCCS. On ground tests suggest that this system is feasible.

Animals↗

Research on Biolab, a multi-user facility for APM.

A study carried out by a team of seven scientists appointed by ESA resulted in the design of a biological laboratory "Biolab" for Columbus APM. The basis for the study were four pre-Phase A studies performed by industry on the assumption that 15 racks would be available to biology and biotechnology in the APM. Due to the constraints newly imposed by the Columbus project, only five racks are now allocated. The tasks of the Biolab scientific team were: (i) to define the scientific objectives of biological research in Columbus; (ii) to review the requirements of the industrial studies; and (iii) to design a multi-purpose facility compatible with the present constraints and satisfying the requirements of the biological investigations considered in the four studies. The Biolab team was able to define a facility capable of accommodating in five racks the following biological objects: small plants (up to 40 cm), insects like drosophila, frog eggs, single cells from animals, bacteria, slime molds and protozoa, as well as human physiology, but restricted to general diagnostic needs. The Biolab facility includes instruments and devices providing the capacity of holding and/or growing the organisms as well as to perform basic experimentation and a minimum essential diagnostic inflight. Within the growth unit the growth chambers/incubators are exchangeable, permitting the use of growth chambers of different sizes. The temperature will be adjustable to the requirements of the objects under investigation, i.e. either 20 or 37 degrees C. Thus a considerable level of flexibility will permit to investigate a broad spectrum of living systems.

Animals↗

Dynamic cell culture system: a new cell cultivation instrument for biological experiments in space.

The prototype of a miniaturized cell cultivation instrument for animal cell culture experiments aboard Spacelab is presented (Dynamic cell culture system: DCCS). The cell chamber is completely filled and has a working volume of 200 microliters. Medium exchange is achieved with a self-powered osmotic pump (flowrate 1 microliter h-1). The reservoir volume of culture medium is 230 microliters. The system is neither mechanically stirred nor equipped with sensors. Hamster kidney (Hak) cells growing on Cytodex 3 microcarriers were used to test the biological performance of the DCCS. Growth characteristics in the DCCS, as judged by maximal cell density, glucose consumption, lactic acid secretion and pH, were similar to those in cell culture tubes.

Animals↗

Cultivation of single cells in space.

The purpose of this review is to present an updated and comprehensive analysis of the experiments with single cells performed in space. Especially the results of the investigations performed in Biorack on the D-1 mission clearly show that important cellular functions are changing in microgravity. Cell proliferation, differentiation, metabolism, membrane properties, and cytoplasmic streaming underwent significant alteration during exposure to space flight conditions in a variety of single cells cultures spanning from bacteria to mammalian cells. These findings open new and interesting perspectives to basic and applied research in microgravity. The focus of this paper is on the cultivation of mammalian cells in space laboratories and on the related instrumentation. While Biorack is a useful and efficient instrument for simple studies in Spacelab, the development of new facilities like incubators with automated fixation devices as well as of more complex bioreactors is strongly recommended.

Animals↗