Cytogenetic examination of cosmonauts for space radiation exposure estimation

Purpose

To evaluate radiation induced chromosome aberration frequency in peripheral blood lymphocytes of cosmonauts who participated in flights on Mir Orbital Station and ISS (International Space Station).

Materials and methods

Cytogenetic examination which has been performed in the period 1992–2008 included the analysis of chromosome aberrations using conventional Giemsa staining method in 202 blood samples from 48 cosmonauts who participated in flights on Mir Orbital Station and ISS.

Results

Space flights led to an increase of chromosome aberration frequency. Frequency of dicentrics plus centric rings (Dic+Rc) depend on the space flight duration and accumulated dose value. After the change of space stations (from Mir Orbital Station to ISS) the radiation load of cosmonauts based on data of cytogenetic examination decreased. Extravehicular activity also adds to chromosome aberration frequency in cosmonauts’ blood lymphocytes. Average doses after the first flight, estimated by the frequency of Dic+Rc, were 227 and 113 mGy Eq for long-term flights (LTF) and 107 and 53 mGy Eq for short-term flights (STF).

Conclusion

Cytogenetic examination of cosmonauts can be applied to assess equivalent doses.     

Lifetime total radiation risk of cosmonauts for orbital and interplanetary flights.

This paper presents results of calculations of total radiation risk for cosmonauts over their lifetimes and assessments of possible shortening of life expectancy on the basis of generalized doses calculated for cosmonauts after a long term interplanetary and orbital space missions on "MIR" station and International Space Station with the use of mathematical expressions coming from a model of change mortality rate of mammals after irradiation. Tumor risk assessments for cosmonauts over lifetime after flights are also given. Dependences of the delayed radiation consequences mentioned above on flight duration, spacecraft shielding thicknesses, solar activity and cosmonauts' age are analyzed.     

Biological dosimetry in Russian and Italian astronauts.

Large uncertainties are associated with estimates of equivalent dose and cancer risk for crews of long-term space missions. Biological dosimetry in astronauts is emerging as a useful technique to compare predictions based on quality factors and risk coefficients with actual measurements of biological damage in-flight. In the present study, chromosomal aberrations were analyzed in one Italian and eight Russian cosmonauts following missions of different duration on the MIR and the international space station (ISS). We used the technique of fluorescence in situ hybridization (FISH) to visualize translocations in chromosomes 1 and 2. In some cases, an increase in chromosome damage was observed after flight, but no correlation could be found between chromosome damage and flight history, in terms of number of flights at the time of sampling, duration in space and extra-vehicular activity. Blood samples from one of the cosmonauts were exposed in vitro to 6 MeV X-rays both before and after the flight. An enhancement in radiosensitivity induced by the spaceflight was observed.     

Chromosomal aberrations induced by high-energy iron ions with shielding.

Biophysical models are commonly used to evaluate the effectiveness of shielding in reducing the biological damage caused by cosmic radiation in space flights. To improve and validate these codes biophysical experiments are needed. We have measured the induction of chromosomal aberrations in human peripheral blood lymphocytes exposed in vitro to 500 MeV/n iron ion beams (dose range 0.1-1 Gy) after traversing shields of different material (lucite, aluminium, or lead) and thickness (0-11.3 g/cm2). For comparison, cells were exposed to 200 MeV/n iron ions and to X-rays. Chromosomes were prematurely condensed by a phosphatase inhibitor (calyculin A) to avoid cell-cycle selection produced by the exposure to high-LET heavy-ion beams. Aberrations were scored in chromosomes 1, 2, and 4 following fluorescence in situ hybridization. The fraction of aberrant lymphocytes has been evaluated as a function of the dose at the sample position, and of the fluence of primary 56Fe ions hitting the shield. The influence of shield thickness on the action cross-section for the induction of exchange-type aberrations has been analyzed, and the dose average-LET measured as a function of the shield thickness. These preliminary results prove that the effectiveness of heavy ions is modified by shielding, and the biological damage is dependent upon shield thickness and material.     

Tissue equivalent detector data obtained recently on Mir space station. Comparison with Solid State Detector data

Tissue equivalent and Solid State Detector (SSD) measurements of the radiation environment inside the Mir space station were performed during the Antares mission in 1992 and long period after it. Interesting results about radiation measurements show (a) the South Atlantic Anomaly (SAA) crossing, (b) the increase of radiation near the poles and (c) the effects of solar eruptions (the most important one occurring in early November 1992). These data give also information about the dose and the quality factor of the radiation received by the cosmonauts during different missions.     

Chromosome aberrations in astronauts

A review of currently available data on in vivo induced chromosome damage in the blood lymphocytes of astronauts proves that cytogenetic biodosimetry analyses on blood collected within a week or two of return from space provides a reliable estimate of equivalent radiation dose and risk after protracted exposure to space radiation of a few months or more. Recent studies indicate that biodosimetry estimates from single spaceflights lie within the range expected from physical dosimetry and biophysical models, but very large uncertainties are associated with single individual measurements and the total sample population remains low. Retrospective doses may be more difficult to estimate because of the fairly rapid time-dependent loss of “stable” aberrations in blood lymphocytes. Also, biodosimetry estimates from individuals who participate in repeated missions, or very long (interplanetary) missions, may be complicated by an adaptive response to space radiation and/or changes in lymphocyte survival and repopulation. A discussion of published data is presented and specific issues related to space radiation biodosimetry protocols are discussed.     

Radiation risk of the crew members of the expeditions on the "MIR" station during the 22nd solar activity cycle.

The radiation risk at the end of the flight was calculated for the members of the main expeditions on the "Mir" station. It was based on the absorbed dose dynamics data measured by the board dosimeter. The radiation damage models created for standards of the radiation safety of the space flights were used in the calculations. The analysis of the obtained values of the risk and its dynamics for some cosmonauts are presented in the topic. The risk values delta P are close to the limited levels given by equation of delta P = 0.6 x 10 x T(-4), [this equation appears also as delta RHrad = 0.6 x 10(-4) x T later in the text] where T--is flight duration in months.     

Biodosimetry of heavyions by interphase chromosome painting

We report measurements of chromosomal aberrations in peripheral blood lymphocytes from cancer patients undergoingradiotherapy treatment. Patients with cervix or esophageal cancer were treated with 10 MV X-rays produced at a LINAC accelerator, or high-energy carbon ions produced at the HIMAC accelerator at the National Institute for Radiological Sciences (NIRS) in Chiba. Blood samples were obtained before, during, and after the radiation treatment. Chromosomes were prematurely condensed by incubation in calyculin A. Aberrations in chromosomes 2 and 4 were scored after fluorescence in situ hybridization with whole-chromosome probes. Pre-treatment samples were exposed in vitro to X-rays, individual dose-response curves for the induction of chromosomal aberrations were determined, and used as calibration curves to calculate the effective whole-body dose absorbed during the treatment. This calculated dose, based on the calibration curve relative to the induction of reciprocal exchanges, has a sharp increase after the first few fractions of the treatment, then saturates at high doses. Although carbon ions are 2–3 times more effective than X-rays in tumor sterilization, the effective dose was similar to that of X-ray treatment. However, the frequency of complex-type chromosomal exchanges was much higher for patients treated with carbon ions than X-ray.     

In vivo and in vitro measurements of complex-type chromosomal exchanges induced by heavy ions.

Heavy ions are more efficient in producing complex-type chromosome exchanges than sparsely ionizing radiation, and this can potentially be used as a biomarker of radiation quality. We measured the induction of complex-type chromosomal aberrations in human peripheral blood lymphocytes exposed in vitro to accelerated H-, He-, C-, Ar-, Fe- and Au-ions in the LET range of approximately 0.4-1400 keV/micrometers. Chromosomes were analyzed either at the first post-irradiation mitosis, or in interphase, following premature condensation by phosphatase inhibitors. Selected chromosomes were then visualized after FISH-painting. The dose-response curve for the induction of complex-type exchanges by heavy ions was linear in the dose-range 0.2-1.5 Gy, while gamma-rays did not produce a significant increase in the yield of complex rearrangements in this dose range. The yield of complex aberrations after 1 Gy of heavy ions increased up to an LET around 100 keV/micrometers, and then declined at higher LET values. When mitotic cells were analyzed, the frequency of complex rearrangements after 1 Gy was about 10 times higher for Ar- or Fe- ions (the most effective ions, with LET around 100 keV/micrometers) than for 250 MeV protons, and values were about 35 times higher in prematurely condensed chromosomes. These results suggest that complex rearrangements may be detected in astronauts' blood lymphocytes after long-term space flight, because crews are exposed to HZE particles from galactic cosmic radiation. However, in a cytogenetic study of ten astronauts after long-term missions on the Mir or International Space Station, we found a very low frequency of complex rearrangements, and a significant post-flight increase was detected in only one out of the ten crewmembers. It appears that the use of complex-type exchanges as biomarker of radiation quality in vivo after low-dose chronic exposure in mixed radiation fields is hampered by statistical uncertainties.     

Simultaneous exposure of mammalian cells to heavy ions and X-rays.

Crews of space missions are exposed to a mixed radiation field, including sparsely and densely ionizing radiation. To determine the biological effectiveness of mixed high-/low-LET radiation fields, mammalian cells were exposed in vitro simultaneously to X-rays and heavy ions, accelerated at the HIMAC accelerator. X-ray doses ranged from 1 to 11 Gy. At the same time, cells were exposed to either 40Ar (550 MeV/n, 86 keV/micrometers), 28Si (100 MeV/n, 150 keV/micrometers), or 56Fe (115 MeV/n, 442 keV/micrometers) ions. Survival was measured in hamster V79 fibroblasts. Structural aberrations in chromosome 2 were measured by chemical-induced premature chromosome condensation combined with fluorescence in situ hybridization in isolated human lymphocytes. For argon and silicon experiments, measured damage in the mixed radiation field was consistent with the value expected using an additive function for low- and high-LET separated data. A small deviation from a simple additive function is observed with very high-LET iron ions combined to X-rays.     

Space Environment Prediction for SZ-4 and SZ-5

The Space Environment Prediction Center (SEPC) of the Center for Space Science and Applied Research of the Chinese Academy of Sciences (CSSAR, CAS)took on the mission of offering the space environment parameters which may be of use to the safety of manned spacecraft. In order to complete the space environment safety guarantee mission for SZ-4 and SZ-5, SEPC improved the space environment monitoring system, database system, prediction result display system, prediction implementation system, etc. For guaranteeing the safety of the airship and cosmonaut in the first manned SZ-5, flying experiment mission,SEPC developed the software for analyzing radiation dose and early-warning software for large debris collision with SZ-5. Three months before the flights of SZ-4 and SZ-5, SEPC began to predict the safe launch period in view of the space environment, and offered timely and valid reference opinions for selecting the safety period. Especially during the mission of SZ-5, SEPC analyzed the space high-energy environment in a pre-arranged orbit and abnormal orbit andevaluated the radiation dose which cosmonauts may encounter in space. The evaluation offered an important reference for cosmonaut safety and decisionmaking in the SZ-5 mission. The calculation of the distribution of large debris and the collision risk assessment at different orbit entry times for SZ-5 provided an important base for the superior department to make flight decisions.     

Role of chromosome instability in long term effect of manned-space missions.

Astronauts are exposed to heavy ions during space missions and heavy ion induced-chromosome damages have been observed in their lymphocytes. This raises the problem of the consequence of longer space flights. Recent studies show that some alterations can appear many cell generations after the initial radiation exposure as a delayed genomic instability. This delayed instability is characterized by the accumulation of cell alterations leading to cell transformation, delayed cell death and mutations. Chromosome instability was shown in vitro in different model systems (Sabatier et al., 1992; Marder and Morgan, 1993, Kadhim et al., 1994 and Holmberg et al., 1993, 1995). All types of radiation used induce a chromosome instability, however, heavy ions cause the most damage. The period of chromosome instability followed by the formation of clones with unbalanced karyotypes seems to be shared by cancer cells. The shortening of telomere sequences leading to the formation of telomere fusions is an important factor in the appearance of this chromosome instability.     

Chromosome aberrations as biomarkers of radiation exposure: modelling basic mechanisms.

The space radiation environment is a mixed field consisting of different particles having different energies, including high charge and energy (HZE) ions. Conventional measurements of absorbed doses may not be sufficient to completely characterise the radiation field and perform reliable estimates of health risks. Biological dosimetry, based on the observation of specific radiation-induced endpoints (typically chromosome aberrations), can be a helpful approach in case of monitored exposure to space radiation or other mixed fields, as well as in case of accidental exposure. Furthermore, various ratios of aberrations (e.g. dicentric chromosomes to centric rings and complex exchanges to simple exchanges) have been suggested as possible fingerprints of radiation quality, although all of them have been subjected to some criticisms. In this context a mechanistic model and a Monte Carlo code for the simulation of chromosome aberration induction were developed. The model, able to provide dose-responses for different aberrations (e.g. dicentrics, rings, fragments, translocations, insertions and other complex exchanges), was further developed to assess the dependence of various ratios of aberrations on radiation quality. The predictions of the model were compared with available data, whose experimental conditions were faithfully reproduced. Particular attention was devoted to the scoring criteria adopted in different laboratories and to possible biases introduced by interphase death and mitotic delay. This latter aspect was investigated by taking into account both metaphase data and data obtained with Premature Chromosome Condensation (PCC).     

Cosmic radiation exposure of aircraft occupants on simulated high-latitude flights during solar proton events from 1 January 1986 through 1 January 2008

From 1 January 1986 through 1 January 2008, GOES satellites recorded 170 solar proton events. For 169 of these events, we estimated effective and equivalent dose rates and doses of galactic cosmic radiation (GCR) and solar cosmic radiation (SCR), received by aircraft occupants on simulated high-latitude flights. Dose rate and dose estimates that follow are for altitudes 30, 40, 50, and 60 kft, in that order.     

Problems and conception of ensuring radiation safety during Mars missions.

The Mars mission differs from near-Earth manned space flights by radiation environment and duration. The importance of effective using the weight of the spacecraft increases greatly because all the necessary things for the mission must be included in its starting weight. For this reason the development of optimal systems of radiation safety ensuring (RSES) acquires especial importance. It is the result of sharp change of radiation environment in the interplanetary space as compared to the one in the near-Earth orbits and significant increase of the interplanetary flight duration. The demand of a harder limitation of unfavorable factors effects should lead to radiation safety (RS) standards hardening. The main principles of ensuring the RS of the Mars mission (optimizing, radiation risk, ALARA) and the conception of RSES, developed on the basis of the described approach and the experience obtained during orbital flights are presented in the report. The problems that can impede the ensuring of the crew members' RS are also given here.     

Human response to high-background radiation environments on Earth and in space

The main long-term objective of the space exploration program is the colonization of the planets of the Solar System. The high cosmic radiation equivalent dose rate represents an inescapable problem for the safe establishment of permanent human settlements on these planets. The unshielded equivalent dose rate on Mars ranges between 100 and 200 mSv/year, depending on the Solar cycle and altitude, and can reach values as high as 360 mSv/year on the Moon. The average annual effective dose on Earth is about 3 mSv, nearly 85% of which comes from natural background radiation, reduced to less than 1 mSv if man-made sources and the internal exposure to Rn daughters are excluded. However, some areas on Earth display anomalously high levels of background radiation, as is the case with thorium-rich monazite bearing sand deposits where values 200–400 times higher than the world average can be found. About 2% of the world’s population live above 3 km and receive a disproportionate 10% of the annual effective collective dose due to cosmic radiation, with a net contribution to effective dose by the neutron component which is 3–4 fold that at sea level. Thus far, epidemiological studies have failed to show any adverse health effects in the populations living in these terrestrial high-background radiation areas (HBRA), which provide an unique opportunity to study the health implications of an environment that, as closely as possibly achievable on Earth, resembles the chronic exposure of future space colonists to higher-than-normal levels of ionizing radiation. Chromosomal aberrations in the peripheral blood lymphocytes from the HBRA residents have been measured in several studies because chromosomal damage represents an early biomarker of cancer risk. Similar cytogenetic studies have been recently performed in a cohort of astronauts involved in single or repeated space flights over many years. The cytogenetic findings in populations exposed to high dose-rate background radiation on Earth or in space will be discussed.     

Biologically weighted measurement of UV radiation in space and on Earth with the biofilm technique.

Biological dosimetry has provided experimental proof of the high sensitivity of the biologically effective UVB doses to changes in atmospheric ozone and has thereby confirmed the predictions from model calculations. The biological UV dosimeter 'biofilm' whose sensitivity is based on dried spores of B. subtilis as UV target weights the incident UV radiation according to its DNA damaging potential. Biofilm dosimetry was applicated in space experiments as well as in use in remote areas on Earth. Examples are long-term UV measurements in Antarctica, measurements of diurnal UV profiles parallel in time at different locations in Europe, continuous UV measurements in the frame of the German UV measurement network and personal UV dosimetry. In space biofilms were used to determine the biological efficiency of the extraterrestrial solar UV, to simulate the effects of decreasing ozone concentrations and to determine the interaction of UVB and vitamin D production of cosmonauts in the MIR station.     

Improved environment radiation models

The two components of the space radiation environment, galactic cosmic rays and solar energetic particles, are of special importance for the planning of space missions and designing space vehicles for flights in the inner heliosphere. There is a constant need for developing and updating the models for calculating the fluxes of these particles for purposes of forecasting radiation conditions anticipated for future flights, including missions to the Moon and Mars.     

Radiation situation determining the possibility of a manned flight to Mars and back.

Possible manned flights toward Mars are discussed from the viewpoint of radiation hazard. A standard situation is considered for the fast two times crossing of the Earth radiation belts. The flight to Mars is shown to be practically impossible without a special system of radiation shelters, because of the effect of penetrating galactic and solar radiations which are responsible for almost maximum permissible doses. But even in case there were radiation shelters on board the spacecraft their flights are undesirable in the periods of maximum and minimum solar activity. It would obviously be worthwhile to schedule Martian flights for intervals in between minima and maxima of 11-year cycles of solar activity when primary cosmic rays levels are considerable reduced and flare activity is not yet sufficiently high. It should be mentioned that it would not be easy to select such allowed intervals. Further studies of that aspect are discussed.