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At least 37 records · Page 2

High-LET radiation-induced aberrations in prematurely condensed G2 chromosomes of human fibroblasts

PURPOSE: To determine the number of initial chromatid breaks induced by low- or high-LET irradiations, and to compare the kinetics of chromatid break rejoining for radiations of different quality. MATERIAL AND METHODS: Exponentially growing human fibroblast cells AG1522 were irradiated with gamma-rays, energetic carbon (290MeV/u), silicon (490MeV/u) and iron (200 and 600 MeV/u). Chromosomes were prematurely condensed using calyculin A. Chromatid breaks and exchanges in G2 cells were scored. PCC were collected after several post-irradiation incubation times, ranging from 5 to 600 min. RESULTS: The kinetics of chromatid break rejoining following low- or high-LET irradiation consisted of two exponential components representing a rapid and a slow time constant. Chromatid breaks decreased rapidly during the first 10min after exposure, then continued to decrease at a slower rate. The rejoining kinetics were similar for exposure to each type of radiation. Chromatid exchanges were also formed quickly. Compared to low-LET radiation, isochromatid breaks were produced more frequently and the proportion of unrejoined breaks was higher for high-LET radiation. CONCLUSIONS: Compared with gamma-rays, isochromatid breaks were observed more frequently in high-LET irradiated samples, suggesting that an increase in isochromatid breaks is a signature of high-LET radiation exposure.

Non-NASA Center↗

LETS: Lunar Environments Test System

The Environmental Effects Branch (EM50) at the Marshall Space Flight Center has developed a unique capability within the agency, namely the Lunar Environment Test System (LETS). LETS is a cryo-pumped vacuum chamber facility capable of high vacuum (10-7 Torr). LETS is a cylindrical chamber, 30 in. (0.8 m) diameter by 48 in. (1.2 m) long thermally controlled vacuum system. The chamber is equipped with a full array of radiation sources including vacuum ultraviolet, electron, and proton radiation. The unique feature of LETS is that it contains a large lunar simulant bed (18 in. x 40 in. x 6 in.) holding 75 kg of JSC-1a simulant while operating at a vacuum of 10-7 Torr. This facility allows three applications: 1) to study the charging, levitation and migration of dust particles, 2) to simulate the radiation environment on the lunar surface, and 3) to electrically charge the lunar simulant enhancing the attraction and adhesion of dust particles to test articles more closely simulating the lunar surface dust environment. LETS has numerous diagnostic instruments including TREK electrostatic probes, residual gas analyzer (RGA), temperature controlled quartz crystal microbalance (TQCM), and particle imaging velocimeter (PIV). Finally, LETS uses continuous Labview data acquisition for computer monitoring and system control.

Vaughn, Jason A.↗

Distribution of Chromosome Breakpoints in Human Epithelial Cells Exposed to Low- and High-LET Radiations

The advantage of the multicolor banding in situ hybridization (mBAND) technique is not only its ability to identify simultaneously both inter- and intrachromosome exchanges, but also the ability to measure the breakpoint location along the length of the chromosome in a precision that is unmatched with other traditional banding techniques. Breakpoints on specific regions of a chromosome have been known to associate with specific cancers. The breakpoint distribution in cells after low- and high-LET radiation exposures will also provide the data for biophysical modeling of the chromatin structure, as well as the data for the modeling the formation of radiation-induced chromosome aberrations. In a series of experiments, we studied low- and high-LET radiation-induced chromosome aberrations using the mBAND technique with chromosome 3 painted in 23 different colored bands. Human epithelial cells (CH1 84B5F5/M10) were exposed in vitro to Cs- 137 rays at both low and high dose rates, secondary neutrons with a broad energy spectrum at a low dose rate and 600 MeV/u Fe ions at a high dose rate. The data of both inter- and intrachromosome aberrations involving the painted chromosome have been reported previously. Here we present data of the location of the chromosome breaks along the length of chromosome 3 in the cells after exposures to each of the four radiation scenarios. In comparison to the expected breakpoint distribution based on the length of the bands, the observed distribution appeared to be non-random for both the low- and high-LET radiations. In particular, hot spots towards both ends of the chromosome were found after low-LET irradiations of either low or high dose rates. For both high-LET radiation types (Fe ions and neutrons), the breakpoint distributions were similar, and were much smoother than that for low-LET radiation. The dependence of the breakpoint distribution on the radiation quality requires further investigations.

Hada, Megumi↗

The 'Pro-Drug' RibCys Decreases The Mutagenicity of High LET Radiation in Cultured Mammalian Cells

We have initiated studies aimed at reducing the mutational effects of high LET radiation such as Fe-56 ions and C-12 ions with certain drugs. The mutagenicity of high LET (143 keV/micrometer) Fe-56 or C-12 ions (LET = 100 keV/micrometer) was quantified at the CD59 locus of human-hamster hybrid AL cells. RibCys [2,S)-D-ribo-(1',2',3',4'- Tetrahydroxybutyl)-thiazolidine-4(R)-ca riboxylic acid], formed by condensation of L-cysteine with D-ribose, is designed so that the sulfhydryl amino acid L-cysteine is released intracellularly via nonenzymatic ring opening and hydrolysis leading to increased levels of glutathione (GSH). RibCys (4 or 10 mM), present during irradiation and a few hours post-irradiation, significantly decreased the yield of CD59(-) mutants induced by radiation. RibCys did not affect the clonogenic survival of irradiated cells, nor was it mutagenic itself. These results, together with the minimal side effects reported in mice and pigs, indicate that RibCys may be useful, perhaps even when used prophylactically, in reducing the load of mutations created by high LET radiation in astronauts or other exposed individuals. RibCys is an attractive drug that may reduce the risk of carcinogenesis in people exposed to high LET radiation.

Lenarczyk, M.↗

Differential Processing of Low and High LET Radiation Induced DNA Damage: Investigation of Switch from ATM to ATR Signaling

The members of the phosphatidylinositol kinase-like kinase family of proteins namely ataxia-telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR) are directly responsible for the maintenance of genomic integrity by mounting DDR through signaling and facilitating the recruitment of repair factors at the sites of DNA damage along with coordinating the deployment of cell cycle checkpoints to permit repair by phosphorylating Checkpoint kinase Chk1, Chk2 and p53. High LET radiation from GCR (Galactic Cosmic Rays) consisting mainly of protons and high energy and charged (HZE) particles from SPE (Solar Particle Event) pose a major health risk for astronauts on their space flight missions. The determination of these risks and the design of potential safeguards require sound knowledge of the biological consequences of lesion induction and the capability of the cells to counter them. We here strive to determine the coordination of ATM and ATR kinases at the break sites directly affecting checkpoint signaling and DNA repair and whether differential processing of breaks induced by low and high LET radiation leads to possible augmentation of swap of these damage sensors at the sites of DNA damage. Exposure of cells to IR triggers rapid autophosphorylation of serine-1981 that causes dimer dissociation and initiates monomer formation of ATM. ATM kinase activity depends on the disruption of the dimer, which allows access and phosphorylation of downstream ATM substrates like Chk2. Evidence suggests that ATM is activated by the alterations in higher-order chromatin structure although direct binding of ATM to DSB ends may be a crucial step in its activation. On the other hand, in case of ATR, RPA (replication protein A)-coated ssDNA (single-stranded DNA) generated as a result of stalled DNA replication or during processing of chromosomal lesions is crucial for the localization of ATR to sites of DNA damage in association with ATR-interacting protein (ATRIP). Although the majority of RPA-coated ssDNA is generally present only during DNA replication, ATR activation in G1 and G2-phase might still require formation of RPA-coated ssDNA, probably initiated by the MRN-CtIP complex and then extended by the Exo1- or BLM-dependent mechanisms at the sites of DSBs. Evidence accumulates that activation of ATM and ATR are oppositely regulated by the length of single stranded overhangs generated at the break sites by processes mentioned above and these stretches of single stranded overhangs hold the clue for ATM to ATR switch at broken DNA ends. We irradiated 82-6hTERT human fibroblast cells with low LET gamma-rays and high LET Fe and Si particles. Preliminary results with cells exposed to 1Gy gamma-rays show that the kinetics of pChk2-pT68 foci formation is comparable to that of gamma-H2AX although they appear to recede quicker. The number and intensity of observed foci reaches a maximum at 30 min and 60 min post IR for Chk2-pT68 and gamma-H2AX foci respectively and all Chk2-pT68 foci colocalize with gamma-H2AX foci. The kinetics of Chk1-pS345 and ATRIP are being determined. Results of Chk2-pT68 foci kinetics was also corroborated by western blot experiments, although phosphorylation was detected as early as 10 min and started receding 30 min post IR with 2Gy of gamma-rays. On the other hand, level of ATR-pS428 reached its maximum between 60 and 120 min and was maintained until the last measured time point of 4 hours post IR as determined by western blotting. Experiments performed with high LET Fe and Si particles will be reported.

Saha, Janapriya↗

Interpreting Space-Mission LET Requirements for SEGR in Power MOSFETs

Single-event gate rupture (SEGR) is a potentially catastrophic failure mechanism of power MOSFETs biased in the off-state. In part due to the severity of-SEGR consequences and in part due to the difficulty of accurated SEGR rate estimation, SEGR mitigation me1hodologies emphasize risk avoidance, using heavy-ion accelerator testing to define safe operating conditions for a surface-incident linear energy transfer {LET). This "safe-operating area" (SOA) within which the device may be biased without experiencing SEGR is then often derated by a prescribed factor to ensure low risk of SEGR. Although research in 1996 indicated that such LET-based SEGR hardness requiremen1s could provide false assurance of safe operation unless one also considered· ion energy], most mission SEGR requirements are still specified in terms of surface-incident LET. Moreover, terrestrial SEGR tests at a given surface-incident LET are limited by the small number of ion species and energies available at heavy-ion accelerators. In comparison, the on-orbit radiation environment is composed of all of the naturally-occurring elements with peak fluxes at nearly GeV/nucleon energies. The primary objective of this study is to examine whether typical derating of high-energy heavy-ion accelerator test data bounds the risk for SEGR from higher-energy on-orbit ions with the mission LET requirement.

Lauenstein, Jean-Marie↗

Comparison of Geant4-DNA and RITRACKS/RITCARD: Microdosimetry, Nanodosimetry and DNA Damage Predictions for Low to High LET Ions

Low linear energy transfer (LET) ionizing radiations induce homogeneously distributed DNA damage at the cellular scale, while high LET ionizing radiations are well known to create complex biological damage including clustered double strand breaks (DSB). Clustered DSB are critical events that are difficult to repair, leading to increased biological outcomes compared to low LET radiation exposure. Recent studies have shown that DNA compaction, which can be actively modified by the cell to allow DNA repair, influences the yield of DSB [1]. The goal of this work is to determine the impact of DNA compaction on chromosome aberration formation, a biomarker of radiation-induced cancer risk. The RITRACKS track structure code [2], together with the RITCARD code [3], allow one to simulate the transport of high LET ions, calculate DNA damage and repair, and chromosome aberration formation, but does not include detailed geometrical models of DNA. The Geant4-DNA toolkit provides such detailed models with different levels of DNA compaction [1,4], but does not model chromosome aberration formation. In this work, these highly detailed DNA geometries are used to compute DNA breaks with Geant4-DNA and combine the results with the RITCARD tool to compute chromosome aberrations. As a first step, we compared the transport models used in RITRACKS and Geant4-DNA and newly released heavy ion model transport (G4DNARuddIonisationExtendedModel Geant4 v11.2) to ensure consistency between the physical stage of the two radiation transport codes. We compared energy deposition at the micrometric and nanometric scales for different ions (H 250 MeV, H 150 Me, He 250 MeV/n, C 290 MeV/n, O 350 MeV/n, O 55 MeV/n, Si 170MeV/n, Ti 300 MeV/n, Ti 300 MeV/n, Fe 600 MeV/n, Fe 450 MeV/n and Fe 300 MeV/n) with LET ranging from ~0.4 keV/μm up to 235 keV/μm. Excellent agreement is found for both microdosimetric and nanodosimetric spectra for all ion types and energies. The calculation of DNA breaks and their complexity with the Geant4-DNA is an ongoing work, and comparison with RITCARD will be presented based on result availability at the time of the conference. [1] Tang et al. (2019). Med. Phys., 46(3), 1501-1511. [2] Plante et al. (2008). NJP, 10(12), 125020. [3] Plante et al. (2019). Rad. Res., 192(3), 282-298. [4] Incerti et al. (2018). Med. Phys., 45(8), e722-e739.

Floriane Poignant↗

A Review: Some biological effects of high LET radiations

There are qualitative and quantitative differences in the biological damage observed after exposure to high LET radiation as compared to that caused by low LET radiations. This review is concerned with these differences, which are ultimately reflected at the biochemical, cellular and even whole animal levels. In general, high LET radiations seem to produce biochemical damage which is more severe and possibly less repairable. Experimental data for those effects are presented in terms of biochemical RBE's with consideration of both early and late manifestations. An LET independent process by which significant biochemical damage may result from protons, neutrons and negative pion mesons is discussed.

Wiley, A., Jr.↗

LET spectra measurements from the STS-35 CPDs

Linear energy transfer (LET) spectra derived form automated track analysis system (ATAS) track parameter measurements for crew passive dosimeters (CPD's) flown with the astronauts on STS-35 are plotted. The spread between the seven individual spectra is typical of past manual measurements of sets of CPD's. This difference is probably due to the cumulative net shielding variations experienced by the CPD's as the astronauts carrying them went about their activities on the Space Shuttle. The STS-35 mission was launched on Dec. 2, 1990, at 28.5 degrees inclination and 352-km altitude. This is somewhat higher than the nominal 300-km flights and the orbit intersects more of the high intensity trapped proton region in the South Atlantic Anomaly (SAA). However, in comparison with APD spectra measured on earlier lower altitude missions (STS-26, -29, -30, -32), the flux spectra are all roughly comparable. This may be due to the fact that the STS-35 mission took place close to solar maximum (Feb. 1990), or perhaps to shielding differences. The corresponding dose and dose equivalent spectra for this mission are shown. The effect of statistical fluctuations at the higher LET values, where track densities are small, is very noticeable. This results in an increased spread within the dose rate and dose equivalent rate spectra, as compared to the flux spectra. The contribution to dose and dose equivalent per measured track is much greater in the high LET region and the differences, though numerically small, are heavily weighted in the integral spectra. The optimum measurement and characterization of the high LET tails of the spectra represent an important part of the research into plastic nuclear track detector (PNTD) response. The integral flux, dose rate, dose equivalent rate and mission dose equivalent for the seven astronauts are also given.

Source record↗

Summary of LET spectra and dose measurements on ten STS missions

A comparison of linear energy transfer (LET) spectra measurements made with plastic nuclear track detectors (PNTD's) from area passive dosimeters (APD's), was made for ten different STS missions under similar shielding. The results show that integral flux, dose rate and equivalent dose rate values follow a general increase with respect to increasing orbital inclination and altitude but that there are large variations from a simple relationship. This is to be expected since it has been shown that Shuttle attitude variations, combined with the anisotropic particle flux at the South Atlantic Anomaly (SAA), can result in differences of a factor of 2 in dose rate inside the Shuttle (Badhwar et al., 1995). Solar cycle and shielding differences also result in variations in radiation dose between STS missions. Spaceflight dosimeters from the STS missions are also being used in the development of a method for increasing LET spectra measurement accuracy by extending LET measurements to particle tracks of ranges 10-80 microns. Refinements in processing and measurement techniques for the flight PNTD's have yielded increased detection efficiencies for the short tracks when LET spectra determined by using the standard and refined methods were intercompared.

Source record↗

Cosmic ray LET spectra and doses on board Cosmos-2044 biosatellite

Results of the experiments on board Cosmos-2044 (Biosatellite 9) are presented. Various nuclear track detectors (NTD) (dielectric, AgCl-based, nuclear emulsions) were used to obtain the Linear Energy Transfer (LET) spectra inside and outside the satellite. The spectra from the different NTDs have proved to be in general agreement. The results of LET spectra calculations using two different models are also presented. The resultant LET distributions are used to calculate the absorbed and equivalent doses and the orbit-averaged quality factors (QF) of the cosmic rays (CR). Absorbed dose rates inside (approximately 20 g cm (exp -2) shielding) and outside (1 g cm(exp -2) the spacecraft, omitting electrons, were found to be 4.8 and 8.6 mrad d (exp -1), respectively, while the corresponding equivalent doses were 8.8 and 19.7 mrem d(exp -1). The effects of the flight parameters on the total fluence of, and on the dose from the CR particles are analyzed. Integral dose distributions of the detected particles are also determined. The LET values which separate absorbed and equivalent doses into 50% intervals are estimated. The CR-39 dielectric NTD is shown to detect 20-30% of the absorbed dose and 60-70% of the equivalent dose in the Cosmos-2044 orbit. The influence of solar activity phase on the magnitude of CR flux is discussed.

Watts, J. W., Jr.↗

Analysis of unrejoined chromosomal breakage in human fibroblast cells exposed to low- and high-LET radiation

Reported studies of DNA breakage induced by radiation of various qualities have generally shown a higher fraction of unrejoined residual breaks after high-LET exposure. This observation is supported by the argument that high-LET radiation induced DNA breaks that are more complex in nature and, thus, less likely to be repaired. In most cases the doses used in these studies were very high. We have studied unrejoined chromosome breaks by analyzing chromosome aberrations using a fluorescence in situ hybridization (FISH) technique with a combination of whole chromosome specific probes and probes specific for the telomere region of the chromosomes. Confluent human fibroblast cells (AG1522) were irradiated with gamma rays, 490 MeV/nucleon Si, or with Fe ions at either 200 and 500 MeV/nucleon, and were allowed to repair at 37 degrees C for 24 hours after exposure. A chemically induced premature chromosome condensation (PCC) technique was used to condense chromosomes in the G2 phase of the cell cycle. Results showed that the frequency of unrejoined chromosome breaks was higher after high-LET radiation, and the ratio of unrejoined to misrejoined chromosome breaks increased steadily with LET up a peak value at 440 keV/microm.

NASA Discipline Radiation Health↗

Kinetics of chromatid break repair in G2-human fibroblasts exposed to low- and high-LET radiations

The purpose of this study is to determine the kinetics of chromatid break rejoining following exposure to radiations of different quality. Exponentially growing human fibroblast cells AG1522 were irradiated with gamma-rays, energetic carbon (290 MeV/u), silicon (490 MeV/u) and iron (200 MeV/u, 600 MeV/u). Chromosomes were prematurely condensed using calyculin A. Prematurely condensed chromosomes were collected after several post-irradiation incubation times, ranging from 5 to 600 minutes, and the number of chromatid breaks and exchanges in G2 cells were scored. The relative biological effectiveness (RBE) for initial chromatid breaks per unit dose showed LET dependency having a peak at 55 keV/micrometers silicon (2.4) or 80 keV/micrometers carbon particles (2.4) and then decreased with increasing LET. The kinetics of chromatid break rejoining following low- or high-LET irradiation consisted of two exponential components. Chromatid breaks decreased rapidly after exposure, and then continued to decrease at a slower rate. The rejoining kinetics was similar for exposure to each type of radiation, although the rate of unrejoined breaks was higher for high-LET radiation. Chromatid exchanges were also formed quickly.

NASA Center JSC↗

TLD efficiency of 7LiF for doses deposited by high-LET particles

The efficiency of 7 LiF TLDs (TLD-700) in registering dose from high-LET (> or = 10 keV/micrometers) charged particles (relative to 137Cs gamma rays) has been measured for a number of accelerated heavy ions at various particle accelerator facilities. These measured efficiency values have been compared with similar results obtained from the open literature and a dose efficiency function has been fitted to the combined data set. While it was found that the dose efficiency is not only a function of LET, but also of the charge of the incident particle, the fitted function can be used to correct the undermeasured value of dose from exposures made in mixed radiation fields where LET information is available. This LET-dependent dose efficiency function is used in our laboratory in determining total absorbed dose and dose equivalent from combined TLD and CR-39 plastic nuclear track detector measurements.

NASA Discipline Radiation Health↗

Repair of clustered DNA damage caused by high LET radiation in human fibroblasts

It has recently been demonstrated experimentally that DNA damage induced by high LET radiation in mammalian cells is non-randomly distributed along the DNA molecule in the form of clusters of various sizes. The sizes of such clusters range from a few base-pairs to at least 200 kilobase-pairs. The high biological efficiency of high LET radiation for induction of relevant biological endpoints is probably a consequence of this clustering, although the exact mechanisms by which the clustering affects the biological outcome is not known. We discuss here results for induction and repair of base damage, single-strand breaks and double-strand breaks for low and high LET radiations. These results are discussed in the context of clustering. Of particular interest is to determine how clustering at different scales affects overall rejoining and fidelity of rejoining of DNA double-strand breaks. However, existing methods for measuring repair of DNA strand breaks are unable to resolve breaks that are close together in a cluster. This causes problems in interpretation of current results from high LET radiation and will require new methods to be developed.

NASA Discipline Radiation Health↗

Cosmic ray LET spectra and doses on board Cosmos-2044 biosatellite

Results of the experiments on board Cosmos-2044 (Biosatellite 9) are presented. Various nuclear track detectors (NTD) (dielectric, AgCl-based, nuclear emulsions) were used to obtain the LET spectra inside and outside the satellite. The spectra from the different NTDs have proved to be in general agreement. The results of LET spectra calculations using two different models are also presented. The resultant LET distributions are used to calculate the absorbed and equivalent doses and the orbit-averaged quality factors (QF) of the cosmic rays (CR). Absorbed dose rates inside (approximately 20 g cm-2 shielding) and outside (1 g cm-2) the spacecraft, omitting electrons, were found to be 4.8 and 8.6 mrad d-1, respectively, while the corresponding equivalent doses were 8.8 and 19.7 mrem d-1. The effects of the flight parameters on the total fluence of, and on the dose from, the CR particles are analyzed. Integral dose distributions of the detected particles are also determined. The LET values which separate absorbed and equivalent doses into 50% intervals are estimated. The CR-39 dielectric NTD is shown to detect 20-30% of the absorbed dose and 60-70% of the equivalent dose in the Cosmos-2044 orbit. The influence of solar activity phase on the magnitude of CR flux is discussed.

Non-NASA Center↗

Comparison of F ratios generated from interphase and metaphase chromosome damage induced by high doses of low- and high-LET radiation

Although biophysical models predict a difference in the ratio of interchromosomal to intrachromosomal interarm exchanges (F ratio) for low- and high-LET radiations, few experimental data support this prediction. However, the F ratios in experiments to date have been generated using data on chromosome aberrations in samples collected at the first postirradiation mitosis, which may not be indicative of the aberrations formed in interphase after exposure to high-LET radiations. In the present study, we exposed human lymphocytes in vitro to 2 and 5 Gy of gamma rays and 3 Gy of 1 GeV/nucleon iron ions (LET = 140 keV/micrometer), stimulated the cells to grow with phytohemagglutinin (PHA), and collected the condensed chromosomes after 48 h of incubation using both chemically induced premature chromosome condensation (PCC) and the conventional metaphase techniques. The PCC technique used here condenses chromosomes mostly in the G(2) phase of the cell cycle. The F ratio was calculated using data on asymmetrical chromosome aberrations in both the PCC and metaphase samples. It was found that the F ratios were similar for the samples irradiated with low- and high-LET radiation and collected at metaphase. However, for irradiated samples assayed by PCC, the F ratio was found to be 8.2 +/- 2.0 for 5 Gy gamma rays and 5.2 +/- 0.9 for 3 Gy iron ions. The distribution of the aberrations indicated that, in the PCC samples irradiated with iron ions, most of the centric rings occurred in spreads containing five or more asymmetrical aberrations. These heavily damaged cells, which were either less likely to reach mitosis or may reach mitosis at a later time, were responsible for the difference in the F ratios generated from interphase and metaphase analysis after exposure to iron ions.

NASA Discipline Radiation Health↗

Chromosome aberrations induced by high-LET radiations

Measurements of chromosome aberrations in peripheral blood lymphocytes are currently the most sensitive and reliable indicator of radiation exposure that can be used for biological dosimetry. This technique has been implemented recently to study radiation exposures incurred by astronauts during space flight, where a significant proportion of the dose is delivered by high-LET particle exposure. Traditional methods for the assessing of cytogenetic damage in mitotic cells collected at one time point after exposure may not be suitable for measuring high-LET radiation effects due to the drastic cell cycle perturbations and interphase cell death induced by this type of exposure. In this manuscript we review the recent advances in methodology used to study high-LET induced cytogenetic effects and evaluate the use of chemically-induced Premature Chromosome Condensation (PCC) as an alternative to metaphase analysis. Published data on the cytogenetic effects of in vitro exposures of high-LET radiation is reviewed, along with biodosimetry results from astronauts after short or long space missions.

ISS Project↗