Neoplastic cell transformation by high-LET radiation: molecular mechanisms.

Experimental data on molecular mechanisms are essential for understanding the bioeffects of radiation and for developing biophysical models, which can help in determining the shape of dose-response curves at very low doses, e.g., doses less than 1 cGy. Although it has been shown that ionizing radiation can cause neoplastic cell transformation directly, that high-LET heavy ions in general can be more effective than photons in transforming cells, and that the radiogenic cell transformation is a multi-step process [correction of processes], we know very little about the molecular nature of lesions important for cell transformation, the relationship between lethal and transformational damages, and the evolution of initial damages into final chromosomal aberrations which alter the growth control of cells. Using cultured mouse embryo cells (C3H10T1/2) as a model system, we have collected quantitative data on dose-response curves for heavy ions with various charges and energies. An analysis of these quantitative data suggested that two DNA breaks formed within 80 angstroms may cause cell transformation and that two DNA breaks formed within 20 angstroms may be lethal. Through studies with restriction enzymes which produce DNA damages at specific sites, we have found that DNA double strand breaks, including both blunt- and cohesive-ended breaks, can cause cell transformation in vitro. These results indicate that DNA double strand breaks can be important primary lesions for radiogenic cell transformation and that blunt-ended double strand breaks can form lethal as well as transformational damages due to misrepair or incomplete repair in the cell. The RBE-LET relationship is similar for HGPRT gene mutation, chromosomal deletion, and cell transformation, suggesting common lesions may be involved in these radiation effects. The high RBE of high-LET radiation for cell killing and neoplastic cell transformation is most likely related to its effectiveness in producing DNA double strand breaks in mammalian cells. At present the role of oncogenes in radiation cell transformation is unclear.     

K6R4016V1D芯片在低地球轨道发生单粒子效应频次的分析

俄罗斯福布斯-土壤火星探测器于2011年11月9日携带中国首个火星探测器萤火一号进入低地球轨道(LEO),但原定于159min后探测器在轨发动机点火变轨未能实施,最终探测计划失败.俄罗斯航天局研究分析认为,事故最可能是由于宇宙线重离子轰击星载计算机存储器件,导致两台计算机重启所致.但是抗辐射专家对空间辐射粒子会在如此短时间内通过单粒子效应(SEE)导致LEO探测器失效的观点并不认同.本文根据俄罗斯航天局发布的受影响器件信息,通过实验和计算,分析了K6R4016V1D芯片在低地球轨道运行时可能遇到的空间辐射粒子诱发单粒子效应的频次,探讨了单粒子效应导致福布斯-土壤火星探测器失效的可能性.      

Life on Mars? II. Physical restrictions.

The primary physical factors important to life's evolution on a planet include its temperature, pressure and radiation regimes. Temperature and pressure regulate the presence and duration of liquid water on the surface of Mars. The prolonged presence of liquid water is essential for the evolution and sustained presence of life on a planet. It has been postulated that Mars has always been a cold dry planet; it has also been postulated that early mars possessed a dense atmosphere of CO2 (> or = 1 bar) and sufficient water to cut large channels across its surface. The degree to which either of these postulates is true correlates with the suitability of Mars for life's evolution. Although radiation can destroy living systems, the high fluxes of UV radiation on the martian surface do not necessarily stop the origin and early evolution of life. The probability for life to have arisen and evolved to a significant degree on Mars, based on the postulated ranges of early martian physical factors, is almost solely related to the probability of liquid water existing on the planet for at least hundreds of millions to billions of years.     

火星生命探测中一种潜在的生物标志物磷酸盐

地外生命探索是国际上广泛关注的深空探测重要目标之一.中国第一个火星探测器天问一号成功发射,开启了对火星表面形貌、生命迹象等进行科学探索的旅程.作为太阳系中与地球最为相似的星球,火星带给人类无穷的遐想.火星上是否存在生命,未来人类是否可以移民火星,磷作为重要的生命元素,在生命的整个进化过程具有不可替代的作用.磷酸盐可以作为一种潜在的生命标志物,为火星生命探测提供新的思路和线索.      

Cosmic radiation and evolution of life on earth: roles of environment, adaptation and selection.

The role of ionizing radiation in general, and cosmic radiation in particular, in the evolution of organisms on the earth by adaptation and natural selection is considered in a series of questions: (1) Are there times during the evolution of the earth and of life when genetic material could be exposed to heavy ion radiation? (2) Throughout the course of chemical and biological evolution on the earth, what fraction of environmental mutagenesis could be attributable to cosmic and/or solar ionizing radiation? (3) Is ionizing radiation an agent of adaptation or selection, or both? (4) What can the cladistics of the evolution of genetic repair tell us about the global history of genotoxic selection pressures? (5) How much genetic diversity can be attributed to the selection of radiation-damage repair processes?     

Biological space experiments for the simulation of Martian conditions: UV radiation and Martian soil analogues.

The survivability of resistant terrestrial microbes, bacterial spores of Bacillus subtilis, was investigated in the BIOPAN facility of the European Space Agency onboard of Russian Earth-orbiting FOTON satellites (BIOPAN I -III missions). The spores were exposed to different subsets of the extreme environmental parameters in space (vacuum, extraterrestrial solar UV, shielding by protecting materials like artificial meteorites). The results of the three space experiments confirmed the deleterious effects of extraterrestrial solar UV radiation which, in contrast to the UV radiation reaching the surface of the Earth, also contains the very energy-rich, short wavelength UVB and UVC radiation. Thin layers of clay, rock or meteorite material were shown to be only successful in UV-shielding, if they are in direct contact with the spores. On Mars the UV radiation climate is similar to that of the early Earth before the development of a protective ozone layer in the atmosphere by the appearance of the first aerobic photosynthetic bacteria. The interference of Martian soil components and the intense and nearly unfiltered Martian solar UV radiation with spores of B. subtilis will be tested with a new BIOPAN experiment, MARSTOX. Different types of Mars soil analogues will be used to determine on one hand their potential toxicity alone or in combination with solar UV (phototoxicity) and on the other hand their UV protection capability. Two sets of samples will be placed under different cut-off filters used to simulate the UV radiation climate of Mars and Earth. After exposure in space the survival of and mutation induction in the spores will be analyzed at the DLR, together with parallel samples from the corresponding ground control experiment performed in the laboratory. This experiment will provide new insights into the principal limits of life and its adaptation to environmental extremes on Earth or other planets which and will also have implications for the potential for the evolution and distribution of life.     

Planetary Protection and the astrobiological exploration of Mars: Proactive steps in moving forward

Future efforts towards Mars exploration should include a discussion about the effects that the strict application of Planetary Protection policies is having on the astrobiological exploration of Mars, which is resulting in a continued delay in the search for Martian life. As proactive steps in the path forward, here we propose advances in three areas. First, we suggest that a redefinition of Planetary Protection and Special Regions is required for the case of Mars. Particularly, we propose a definition for special places on Mars that we can get to in the next 10–20?years with rovers and landers, where try to address questions regarding whether there is present-day near-surface life on Mars or not, and crucially doing so before the arrival of manned missions. We propose to call those special places “Astrobiology Priority Exploration” regions (APEX regions). Second, we stress the need for the development of robotic tools for the characterization of complex organic compounds as unequivocal signs of life, and particularly new generations of complex organic chemistry and biosignature detection instruments, including advances in DNA sequencing. And third, we advocate for a change from the present generation of SUV-sized landers and rovers to new robotic assets that are much easier to decontaminate such as microlanders: they would be very small with limited sensing capabilities, but there would be many of them available for launch and coordination from an orbiting platform. Implementing these changes will help to move forward with an exploration approach that is much less risky to the potential Mars biosphere, while also being much more scientifically rigorous about the exploration of the “life on Mars” question – a question that needs to be answered both for astrobiological discovery and for learning more definitive lessons on Planetary Protection.     

Early martian environments: the Antarctic and other terrestrial analogs.

The comparability of the early environments of Mars and Earth, and the biological evolution which occurred on early earth, motivates serious consideration of the possibility of an early martian biota. Environments which could have contained this early martian life and which may presently contain evidence of this former life include aquatic, ice, soil, and rock habitats. Several analogs of these potential early martian environments, which can provide useful information in searching for extinct life on Mars, are currently available for study on Earth. These terrestrial analogs include the perennially ice-covered lakes and sandstone rocks in the Polar Deserts of Antarctica, surface of snowfields and glaciers, desert soils, geothermal springs, and deep subsurface environments.     

Planetary protection issues in advance of human exploration of Mars.

Current planetary quarantine considerations focus on robotic missions and attempt a policy of no biological contamination. The presence of humans on Mars, however, will inevitably result in biological contamination and physical alteration of the local environment. The focus of planetary quarantine must therefore shift toward defining and minimizing the inevitable contamination associated with humans. This will involve first determining those areas that will be affected by the presence of a human base, then verifying that these environments do not harbor indigenous life nor provide sites for Earth bacteria to grow. Precursor missions can provide salient information that can make more efficient the planning and design of human exploration missions. In particular, a robotic sample return mission can help to eliminate the concern about returning samples with humans or the return of humans themselves from a planetary quarantine perspective. Without a robotic return the cost of quarantine that would have to be added to a human mission may well exceed the cost of a robotic return mission. Even if the preponderance of scientific evidence argues against the presence of indigenous life, it must be considered as part of any serious planetary quarantine analysis for missions to Mars. If there is life on Mars, the question of human exploration assumes an ethical dimension.     

Radiation issues for piloted Mars mission.

Man is now entering an era of colonizing the moon and exploration of Mars. The crewmembers of a piloted mission to Mars will be exposed to inner belt trapped protons, the outer trapped electrons, and the galactic cosmic radiation. In addition there is always the added risk of acute exposure to a solar particle event. Current radiation risk is estimated using the idea of absorbed dose and ICRP-26, LET-dependent quality factors. In a spacecraft with aluminum walls (2 g cm-2) at solar minimum the calculated dose equivalent is 0.73 Sv for a 406-day mission. Based on the current thinking this leads to an excess cancer mortality in a 35 year male of about 1%. About 75% of the dose equivalent is contributed by HZE particles and target fragments with average quality factors of 10.3 and 20, respectively. The entire concept of absorbed dose, quality factor, and dose equivalent as applied to such missions needs to be reexamined, in light of the fact that less than 50% of the nuclei in the body of the astronaut would have been traversed by a single GCR nuclei in the 406-day mission. Clearly, more biologically relevant information about the effects of heavy ions and target fragments is needed and fluence based risk estimation strategy developed for such long term stays in space.     

Exobiology and future Mars missions: The search for Mars'' earliest biosphere

The primordial Mars may have possessed a thick carbon dioxide atmosphere, with liquid water common on the surface, similar in many ways to the primordial Earth. During this epoch, billions of years ago, the surface of Mars could have been conducive to the origin of life. It is possible that life evolved on Mars to be later eliminated as the atmospheric pressure dropped. Analysis of the surface of Mars for the traces of this early martian biota could provide many insights into the phenomenon of life and its coupling to planetary evolution.     

Radiation biology in space: a critical review.

A short summary of the results of radiobiological studies in space or on respective particles on ground will be given. Among the various types of radiation in space, the effect of heavy ions with high energy (HZE-particles) are most essential. Thus, radiobiology in space concerns mostly to the effect of these particles, in cells and in whole organism. Cell death, mutation and malignant transformation are the relevant endpoints, with can be studied on ground with heavy ions of different energy with suitable accelerators or in space, especially by the BIOSTACK concept. In space, however, the effect of microgravity has to be considered as well and there are hints, that under weightlessness the biological effect of radiation may be enhanced. There are still open questions to be answered concerning radioprotection of man in space. Further experiments are necessary.     

Survival of microorganisms in space: A review

Spores of $\text{Bacillus subtilis}$ were exposed to selected factors of space (vacuum, solar UV radiation, heavy ions of cosmic radiation), and their response was studied after recovery. These investigations were supplemented by ground-based studies under simulated space conditions. The vacuum of space did not inactivate the spores. However, vacuum-induced structural changes in the DNA, and probably in the proteins, caused a supersensitivity to solar UV radiation. This phenomenon is caused by the production of specific photoproducts in DNA and protein, which cannot be removed by normal cellular repair processes. In vegetative bacterial cells, exposed to vacuum, cell dehydration led to damage of the cell membrane, which could be partly repaired during subsequent incubation. The high local effectiveness of the cosmic heavy ions further decreases the chance that spores can survive for any length of time in space. Nonetheless, a spore travelling through space and protected from ultraviolet radiation could possibly survive an interplanetary journey. Such a situation favors panspermia as a possible explanation for the origin of life.     

Cell cycle delays induced by heavy ion irradiation of synchronous mammalian cells.

Cell cycle effects of very high LET particles on synchronous V79 Chinese Hamster cells have been studied in a track segment experiment by means of flow cytometric methods. Cells were irradiated with 10 MeV/u Pb-ions (LET = 13500 keV/micrometers) at an average fluence of 2 particles per cell nucleus, corresponding to a survival level of about 25%. Instantaneous drastic reductions of cell proliferation in all cycle phases have been observed, which affect the cell cycle for at least 50 hours after exposure to heavy ions. These findings are in clear contrast to the results from low LET radiation experiments, where significant delays can only be observed in S-phase and G2M-phase and for comparatively short time intervals of a few hours. Additionally, high LET radiation gives rise to prolonged DNA synthesis bypassing cell division, which leads to cells with DNA content greater than that of G2M-cells.     

γ-ray spectroscopy in Mars orbit during solar proton events

Understanding the evolution of Mars requires determining the composition of the surface and atmosphere of the planet. The European Space Agency’s ExoMars rover mission, which is expected to launch in 2016, is part of the Aurora programme. The instruments on the rover will search for evidence of life on Mars and will map a sub-section of the Martian surface, extracting compositional information. Currently our understanding of the bulk composition (and mineralogy) of Mars relies on orbital data from instruments on-board satellites such as 2001 Mars Odyssey, Mars Reconnaissance Orbiter and Mars Express, in addition to in-situ instrumentation on rovers such as Spirit and Opportunity. γ-ray spectroscopy can be used to determine the composition of Mars, but it has yet to be successfully carried out in-situ on Mars. This study describes some of the results obtained from the γ-ray spectrometer on 2001 Mars Odyssey during solar proton events and discusses whether the increased emissions are useful in γ-ray spectroscopy. The study shows that although increased γ-ray emissions were expected from the Martian surface during a solar proton event, they were not detected from orbit probably due to insufficient signal-to-background. However, this does not preclude the possibility of measuring changes in γ-ray flux corresponding to changes in solar activity on the surface of the planet.     

Life on Mars? I. The chemical environment.

The origin of life at its abiotic evolutionary stage, requires a combination of constituents and environmental conditions that enable the synthesis of complex replicating macromolecules from simpler monomeric molecules. It is very likely that the early stages of this evolutionary process have been spontaneous, rapid and widespread on the surface of the primitive Earth, resulting in the formation of quite sophisticated living organisms within less than a billion years. To what extent did such conditions prevail on Mars? Two companion-papers (Life on Mars? I and II) will review and discuss the available information related to the chemical, physical and environmental conditions on Mars and assess it from the perspective of potential exobiological evolution.     

Dictyostelium discoideum, a lower eukaryote model for the study of DNA repair: implications for the role of DNA-damaging chemicals in the evolution of repair proficient cells.

The evolution of the ability of living cells to cope with stress is crucial for the maintenance of their genetic integrity. Yet low levels of mutation must remain to allow adaptation to environmental changes. The cellular slime mold D. discoideum is a good system for studying molecular aspects of the repair of lethal and mutagenic damage to DNA by radiation and chemicals. The wild-type strains of this soil microorganism are extremely resistant to DNA damaging agents. In nature the amoeboid cells in their replicative stage feed on soil bacteria and are exposed to numerous DNA-damaging chemicals produced by various soil microorganisms. It is probable that the evolution of repair systems in this organism and perhaps in others is a consequence of the necessity to cope with chemical damage which also confers resistance to radiation.     

From mechanisms to risk estimation--bridging the chasm.

We have a considerable amount of work ahead of us to determine the importance of the wealth of new information emerging in the fields of sub-cellular, cellular and tissue biology in order to improve the estimation of radiation risk at low dose and protracted dose-rate. In this paper, we suggest that there is a need to develop models of the specific health effects of interest (e.g., carcinogenesis in specific tissues), which embody as much of the mechanistic (i.e., biological) information as is deemed necessary. Although it is not realistic to expect that every radiation-induced process should or could be included, we can hope that the major factors that shape the time dependence of evolution of damage can be identified and quantified to the point where reasonable estimations of risk can be made. Regarding carcinogenesis in particular, the structure of the model itself plays a role in determining the relative importance of various processes. We use a specific form of a multi-stage carcinogenic model to illustrate this point. We show in a review of the application of this model to lung cancer incidence and mortality in two exposed populations that for both high- and low-LET radiation, there is evidence of an "inverse dose-rate" or protraction effect. This result could be of some considerable importance, because it would imply that risk from protracted exposure even to low-LET radiation might be greater than from acute exposure, an opinion not currently held in the radiation protection community. This model also allows prediction of the evolution of the risk over the lifetimes of the exposed individuals. One inference is that radiation-induced initiation (i.e., the first cellular carcinogenic event(s) occurring in normal tissue after the passage of the radiation) may not be the driving factor in the risk, but more important may be the effects of the radiation on already-initiated cells in the tissue. Although present throughout the length of the exposure, radiation-induced initiation appears to play a dominating role only very late in life, and only for those individuals who began their exposure early in life. These conclusions are very dependent, of course, on the hypotheses embodied in the initiation-promotion-conversion paradigm of carcinogenesis. We suggest that recently identified processes, such as the "bystander effect", might affect initiation, promotion, and malignant conversion in different ways. Finally, the manner in which the quality of radiation affects these processes must be understood in the context of the mixed high- and low-LET radiations that are found in the space environment. Important directions in critical experiment definition are suggested, including a renewed emphasis on well-designed animal experiments over extended periods of time.     

Preliminary results of space experiment: Implications for the effects of space radiation and microgravity on survival and mutation induction in human cells

In view of the concern for the health of astronauts that may one day journey to Mars or the Moon, we investigated the effect that space radiation and microgravity might have on DNA damage and repair. We sent frozen human lymphoblastoid TK6 cells to the International Space Station where they were maintained under frozen conditions during a 134-day mission (14 November 2008 to 28 March 2009) except for an incubation period of 8 days under 1G or μG conditions in a CO₂ incubator. The incubation period started after 100 days during which the cells had been exposed to 54 mSv of space radiation. The incubated cells were then refrozen, returned to Earth, and compared to ground control samples for the determination of the influence of microgravity on cell survival and mutation induction. The results for both varied from experiment to experiment, yielding a large SD, but the μG sample results differed significantly from the 1G sample results for each of 2 experiments, with the mean ratio of μG to 1G being 0.55 for the concentration of viable cells and 0.59 for the fraction of thymidine kinase deficient (TK⁻) mutants. Among the mutants, non-loss of zygosity events (point mutations) were less frequent (31%) after μG incubation than after 1G incubation, which might be explained by the influence of μG on cellular metabolic or physiological function. Additional experiments are needed to clarify the effect of μG interferes on DNA repair.     

Annual solar UV exposure and biological effective dose rates on the Martian surface.

The ultraviolet (UV) environment of Mars has been investigated to gain an understanding of the variation of exposure throughout a Martian year, and link this flux to biological effects and possible survival of organisms at the Martian surface. To gain an idea of how the solar UV radiation varies between different regions, including planned landing sites of two future Mars surface missions, we modelled the total solar UV surface flux throughout one Martian year for two different dust scenarios. To understand the degree of solar UV stress on micro-organisms and/or molecules essential for life on the surface of Mars, we also calculated the biologically effective dose (BED) for T7 and Uracil in relevant wavelength regions at the Martian surface as a function of season and latitude, and discuss the biological survival rates in the presence of Martian solar UV radiation. High T7/Uracil BED ratios indicate that even at high latitudes where the UV flux is significantly reduced, the radiation environment is still hostile for life due to the persisting UV-C component of the flux.