Applicability of composite materials for space radiation shielding of spacecraft.

Energetic ion beam experiments with major space radiation elements, 1H, 4He, 16O, 28Si and 56Fe, have been conducted to investigate the radiation shielding properties of composite materials. These materials are expected to be used for parts and fixtures of space vehicles due to both their mechanical strength and their space radiation shielding capabilities. Low Z materials containing hydrogen are effective for shielding protons and heavy ions due to their high stopping power and large fragmentation cross section per unit mass. The stopping power of the composite materials used in this work is intermediate between that of aluminum and polyethylene, which are typical structural and shielding materials used in space. The total charge-changing cross sections per unit mass, σUM, of the composite materials are 1.3-1.8 times larger than that of aluminum. By replacing conventional aluminum used for spacecraft with commercially available composite (carbon fiber / polyether ether ketone), it is expected that the shielding effect is increased by ∼17%. The utilization of composite materials will help mitigate the space radiation hazard on future deep space missions.

Space Radiation Dosimetry at the Exposure Facility of the International Space Station for the Tanpopo Mission.

Radiation dosimetry was carried out at the exposure facility (EF) and the pressurized module (PM) of the Japanese Kibo module installed in the International Space Station as one study on environmental monitoring for the Tanpopo mission. Three exposure panels and three references including biological and organic samples and luminescence dosimeters were launched to obtain data for different exposure durations during 3 years from May 2015 to July 2018. The dosimeters were equipped with additional shielding materials (0.55, 2.95, and 6.23 g/cm2 mass thickness). The relative dose variation, as a function of shielding mass thickness, was observed and compared with Monte Carlo simulations with respect to galactic cosmic rays (GCRs) and typical solar energetic particles (SEPs). The mean annual dose rates were DEF = 231 ± 5 mGy/year at the EF and DPM = 82 ± 1 mGy/year at the PM during the 3 years. The PM is well shielded, and the GCR simulation indicated that the measured mean dose reduction ratio inside the module (DPM/DEF = 0.35) required ∼26 g/cm2 additional shielding mass thickness. Observed points of the dose reduction tendency could be explained by the energy ranges of protons (10-100 MeV), where the protons passed through, or were absorbed in, the shielding materials of different mass thickness that surrounded dosimeters.

DNA Damage and Survival Time Course of Deinococcal Cell Pellets During 3 Years of Exposure to Outer Space.

The hypothesis called "panspermia" proposes an interplanetary transfer of life. Experiments have exposed extremophilic organisms to outer space to test microbe survivability and the panspermia hypothesis. Microbes inside shielding material with sufficient thickness to protect them from UV-irradiation can survive in space. This process has been called "lithopanspermia," meaning rocky panspermia. We previously proposed sub-millimeter cell pellets (aggregates) could survive in the harsh space environment based on an on-ground laboratory experiment. To test our hypothesis, we placed dried cell pellets of the radioresistant bacteria Deinococcus spp. in aluminum plate wells in exposure panels attached to the outside of the International Space Station (ISS). We exposed microbial cell pellets with different thickness to space environments. The results indicated the importance of the aggregated form of cells for surviving in harsh space environment. We also analyzed the samples exposed to space from 1 to 3 years. The experimental design enabled us to get and extrapolate the survival time course to predict the survival time of Deinococcus radiodurans. Dried deinococcal cell pellets of 500 µm thickness were alive after 3 years of space exposure and repaired DNA damage at cultivation. Thus, cell pellets 1 mm in diameter have sufficient protection from UV and are estimated to endure the space environment for 2-8 years, extrapolating the survival curve and considering the illumination efficiency of the space experiment. Comparison of the survival of different DNA repair-deficient mutants suggested that cell aggregates exposed in space for 3 years suffered DNA damage, which is most efficiently repaired by the uvrA gene and uvdE gene products, which are responsible for nucleotide excision repair and UV-damage excision repair. Collectively, these results support the possibility of microbial cell aggregates (pellets) as an ark for interplanetary transfer of microbes within several years.

Biologic Impact of Different Ultra-Low-Fluence Irradiations in Human Fibroblasts.

In this study, we aimed to evaluate the cellular response of healthy human fibroblasts induced by different types of ultra-low-fluence radiations, including gamma rays, neutrons and high linear energy transfer (LET) heavy ions. NB1RGB cells were pretreated with ultra-low-fluence radiations (~0.1 cGy/7-8 h) of 137Cs gamma rays, 241Am-Be neutrons, helium, carbon and iron ions before being exposed to an X-ray-challenging dose (1.5 Gy). Helium (LET = 2.3 keV/µm), carbon (LET = 13.3 keV/µm) and iron (LET = 200 keV/µm) ions were generated with the Heavy Ion Medical Accelerator in Chiba (HIMAC), Japan. No differences in cell death-measured by colony-forming assay-were observed regardless of the radiation type applied. In contrast, mutation frequency, which was detected through cell transformation into 6-thioguanine resistant clones, was 1.9 and 4.0 times higher in cells pretreated with helium and carbon ions, respectively, compared to cells exposed to X-ray-challenging dose alone. Moreover, cells pretreated with iron ions or gamma-rays showed a mutation frequency similar to cells exposed to X-ray-challenging dose alone, while cells pretreated with neutrons had 0.15 times less mutations. These results show that cellular responses triggered by ultra-low-fluence irradiations are radiation-quality dependent. Altogether, this study shows that ultra-low-fluence irradiations with the same level as those reported in the International Space Station are capable of inducing different cellular responses, including radio-adaptive responses triggered by neutrons and genomic instability mediated by high-LET heavy ions, while electromagnetic radiations (gamma rays) seem to have no biologic impact.

Investigation of shielding material properties for effective space radiation protection.

Geant4 Monte Carlo simulations were carried out to investigate the possible shielding materials of aluminum, polyethylene, hydrides, complex hydrides and composite materials for radiation protection in spacecraft by considering two physical parameters, stopping power and fragmentation cross section. The dose reduction with shielding materials was investigated for Fe ions with energies of 500 MeV/n, 1 GeV/n and 2 GeV/n which are around the peak of the GCR energy spectrum. Fe ions easily stop in materials such as polyethylene and hydrides as opposed to materials such as aluminum and complex hydrides including high Z metals with contain little or no hydrogen. Attenuation of the primary particles in the shielding and fragmentation into more lightly charged and therefore more penetrating secondary particles are competing factors: attenuation acts to reduce the dose behind shielding while fragmentation increases it. Among hydrogenous materials, 6Li10BH4 was one of the more effective shielding materials as a function of mass providing a 20% greater dose reduction compared to polyethylene. Composite materials such as carbon fiber reinforced plastic and SiC composite plastic offer 1.9 times the dose reduction compared to aluminum as well as high mechanical strength. Composite materials have been found to be promising for spacecraft shielding, where both mass and volume are constrained.

Environmental Data and Survival Data of Deinococcus aetherius from the Exposure Facility of the Japan Experimental Module of the International Space Station Obtained by the Tanpopo Mission.

The Tanpopo mission has two objectives: (1) test the panspermia hypothesis and (2) test whether organic compounds may have been transferred to Earth before the origin of life. We developed an exposure panel (EP) designed to expose microbes and organic compounds to the space environment and a capture panel designed to capture high-velocity particles on the International Space Station (ISS) using aerogel contained in an aluminum container. The panels returned after 1 year of exposure at the Exposure Facility of the Japan Experimental Module, ISS. In this communication, we report the measurements of temperature, radiation dosimeter and vacuum ultraviolet dosimeter in the EP, and survival data of Deinococcus aetherius. The environmental data are consistent with survival data of microbes and organic compounds, which will be presented elsewhere in detail.

Low 134Cs/137Cs ratio anomaly in the north-northwest direction from the Fukushima Dai-ichi Nuclear Power Station.

A low 134Cs/137Cs ratio anomaly in the north-northwest (NNW) direction from the Fukushima Dai-ichi Nuclear Power Station (FDNPS) is identified by a new analysis of the 134Cs/137Cs ratio dataset which we had obtained in 2011-2015 by a series of car-borne surveys that employed a germanium gamma-ray spectrometer. We found that the 134Cs/137Cs ratio is slightly lower (0.95, decay-corrected to March 11, 2011) in an area with a length of about 15 km and a width of about 3 km in the NNW direction from the FDNPS than in other directions from the station. Furthermore, the area of this lower 134Cs/137Cs ratio anomaly corresponds to a narrow contamination band that runs NNW from the FDNPS and it is nearly parallel with the major and heaviest contamination band in the west-northwest. The plume trace with a low 134Cs/137Cs ratio previously found by other researchers within the 3-km radius of the FDNPS is in a part of the area with the lower 134Cs/137Cs ratio anomaly that we found. Our result suggests that this lower 134Cs/137Cs ratio anomaly is the area which was contaminated before March 13, 2011 (UTC) in association with the hydrogen explosion of Unit 1 on March 12, 2011 at 06:36 (UTC) and it was less influenced by later subsequent plumes.

Galactic cosmic ray simulation at the NASA Space Radiation Laboratory.

Most accelerator-based space radiation experiments have been performed with single ion beams at fixed energies. However, the space radiation environment consists of a wide variety of ion species with a continuous range of energies. Due to recent developments in beam switching technology implemented at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL), it is now possible to rapidly switch ion species and energies, allowing for the possibility to more realistically simulate the actual radiation environment found in space. The present paper discusses a variety of issues related to implementation of galactic cosmic ray (GCR) simulation at NSRL, especially for experiments in radiobiology. Advantages and disadvantages of different approaches to developing a GCR simulator are presented. In addition, issues common to both GCR simulation and single beam experiments are compared to issues unique to GCR simulation studies. A set of conclusions is presented as well as a discussion of the technical implementation of GCR simulation.

Non-induction of radioadaptive response in zebrafish embryos by neutrons.

In vivo neutron-induced radioadaptive response (RAR) was studied using zebrafish (Danio rerio) embryos. The Neutron exposure Accelerator System for Biological Effect Experiments (NASBEE) facility at the National Institute of Radiological Sciences (NIRS), Japan, was employed to provide 2-MeV neutrons. Neutron doses of 0.6, 1, 25, 50 and 100 mGy were chosen as priming doses. An X-ray dose of 2 Gy was chosen as the challenging dose. Zebrafish embryos were dechorionated at 4 h post fertilization (hpf), irradiated with a chosen neutron dose at 5 hpf and the X-ray dose at 10 hpf. The responses of embryos were assessed at 25 hpf through the number of apoptotic signals. None of the neutron doses studied could induce RAR. Non-induction of RAR in embryos having received 0.6- and 1-mGy neutron doses was attributed to neutron-induced hormesis, which maintained the number of damaged cells at below the threshold for RAR induction. On the other hand, non-induction of RAR in embryos having received 25-, 50- and 100-mGy neutron doses was explained by gamma-ray hormesis, which mitigated neutron-induced damages through triggering high-fidelity DNA repair and removal of aberrant cells through apoptosis. Separate experimental results were obtained to verify that high-energy photons could disable RAR. Specifically, 5- or 10-mGy X-rays disabled the RAR induced by a priming dose of 0.88 mGy of alpha particles delivered to 5-hpf zebrafish embryos against a challenging dose of 2 Gy of X-rays delivered to the embryos at 10 hpf.

G(2)-M phase-correlative bystander effects are co-mediated by DNA-PKcs and ATM after carbon ion irradiation.

Accumulated evidence has shown that radiation-induced bystander effect (RIBE) may have significant implications to the efficiency of radiotherapy. Although cellular radiosensitivity relies on cell cycle status, it is largely unknown how about the relationship between RIBE and cell cycle distribution, much less the underlying mechanism. In the present study, the lung cancer A549 cells were synchronized into different cell cycle phases of G1, S and G2/M and irradiated with high linear energy transfer (LET) carbon ions. By treating nonirradiated cells with the conditioned medium from these irradiated cells, it was found that the G2-M phase cells had the largest contribution to RIBE. Meanwhile, the activity of DNA-PKcs but not ATM was increased in the synchronized G2-M phase cells in spite of both of them were activated in the asynchronous cells after carbon ion irradiation. When the G2-M phased cells were transferred with DNA-PKcs siRNA and ATM siRNA individually or treated with an inhibitor of either DNA-PKcs or ATM before carbon ion irradiation, the RIBE was effectively diminished. These results provide new evidence linking cell cycle to bystander responses and demonstrate that DNA-PKcs and ATM are two associated factors in co-regulating G2-M phase-related bystander effects.

Induction of Non-Targeted Stress Responses in Mammary Tissues by Heavy Ions.

PURPOSE: Side effects related to radiation exposures are based primarily on the assumption that the detrimental effects of radiation occur in directly irradiated cells. However, several studies have reported over the years of radiation-induced non-targeted/ abscopal effects in vivo that challenge this paradigm. There is evidence that Cyclooxygenase-2 (COX2) plays an important role in modulating non-targeted effects, including DNA damages in vitro and mutagenesis in vivo. While most reports on radiation-induced non-targeted response utilize x-rays, there is little information available for heavy ions. METHODS AND MATERIALS: Adult female transgenic gpt delta mice were exposed to an equitoxic dose of either carbon or argon particles using the Heavy Ion Medical Accelerator in Chiba (HIMAC) at the National Institute of Radiological Sciences (NIRS) in Japan. The mice were stratified into 4 groups of 5 animals each: Control; animals irradiated under full shielding (Sham-irradiated); animals receiving whole body irradiation (WBIR); and animals receiving partial body irradiation (PBIR) to the lower abdomen with a 1 x 1 cm2 field. The doses used in the carbon ion group (4.5 Gy) and in argon particle group (1.5 Gy) have a relative biological effectiveness equivalent to a 5 Gy dose of x-rays. 24 hours after irradiation, breast tissues in and out of the irradiated field were harvested for analysis. Induction of COX2, 8-hydroxydeoxyguanosine (8-OHdG), phosphorylated histone H2AX (γ-H2AX), and apoptosis-related cysteine protease-3 (Caspase-3) antibodies were examined in the four categories of breast tissues using immunohistochemical techniques. Analysis was performed by measuring the intensity of more than 20 individual microscopic fields and comparing the relative fold difference. RESULTS: In the carbon ion group, the relative fold increase in COX2 expression was 1.01 in sham-irradiated group (p > 0.05), 3.07 in PBIR (p < 0.05) and 2.50 in WBIR (p < 0.05), respectively, when compared with controls. The relative fold increase in 8-OHdG expression was 1.29 in sham-irradiated (p > 0.05), 11.31 in PBIR (p < 0.05) and 11.79 in WBIR (p < 0.05), respectively, when compared with controls. A similar increase in γ-H2AX expression was found in that, compared to controls, the increase was 1.41 fold in sham-irradiated (p > 0.05), 8.41 in PBIR (p < 0.05) and 10.59 in WBIR (p < 0.05). Results for the argon particle therapy group showed a similar magnitude of changes in the various biological endpoints examined. There was no statistical significance observed in Caspase-3 expression among the 4 groups. CONCLUSIONS: Our data show that both carbon and argon ions induced non-targeted, out of field induction of COX2 and DNA damages in breast tissues. These effects may pose new challenges to evaluate the risks associated with radiation exposure and understanding radiation-induced side effects.

Rescue of Targeted Nonstem-Like Cells from Bystander Stem-Like Cells in Human Fibrosarcoma HT1080.

Cancer stem-like cells (CSCs) have been suggested to be the principal cause of tumor radioresistance, dormancy and recurrence after radiotherapy. However, little is known about CSC behavior in response to clinical radiotherapy, particularly with regard to CSC communication with bulk cancer cells. In this study, CSCs and nonstem-like cancer cells (NSCCs) were co-cultured, and defined cell types were chosen and irradiated, respectively, with proton microbeam. The bidirectional rescue effect in the combinations of the two cell types was then investigated. The results showed that out of all four combinations, only the targeted, proton irradiated NSCCs were protected by bystander CSCs and showed less accumulation of 53BP1, which is a widely used indicator for DNA double-strand breaks. In addition, supplementation with c-PTIO, a specific nitric oxide scavenger, can show a similar effect on targeted NSCCs. These results, showed that the rescue effect of CSCs on targeted NSCCs involves nitric oxide in the process, suggesting that the cellular communication between CSCs and NSCCs may be important in determining the survival of tumor cells after radiation therapy. To our knowledge, this is the first report demonstrating a rescue effect of CSCs to irradiated NSCCs that may help us better understand CSC behavior in response to cancer radiotherapy.

Comparison of cosmic rays radiation detectors on-board commercial jet aircraft.

Aircrew members and passengers are exposed to increased rates of cosmic radiation on-board commercial jet aircraft. The annual effective doses of crew members often exceed limits for public, thus it is recommended to monitor them. In general, the doses are estimated via various computer codes and in some countries also verified by measurements. This paper describes a comparison of three cosmic rays detectors, namely of the (a) HAWK Tissue Equivalent Proportional Counter; (b) Liulin semiconductor energy deposit spectrometer and (c) TIMEPIX silicon semiconductor pixel detector, exposed to radiation fields on-board commercial Czech Airlines company jet aircraft. Measurements were performed during passenger flights from Prague to Madrid, Oslo, Tbilisi, Yekaterinburg and Almaty, and back in July and August 2011. For all flights, energy deposit spectra and absorbed doses are presented. Measured absorbed dose and dose equivalent are compared with the EPCARD code calculations. Finally, the advantages and disadvantages of all detectors are discussed.

The differential role of human macrophage in triggering secondary bystander effects after either gamma-ray or carbon beam irradiation.

The abscopal effect could be an underlying factor in evaluating prognosis of radiotherapy. This study established an in vitro system to examine whether tumor-generated bystander signals could be transmitted by macrophages to further trigger secondary cellular responses after different irradiations, where human lung cancer NCI-H446 cells were irradiated with either γ-rays or carbon ions and co-cultured with human macrophage U937 cells, then these U937 cells were used as a bystander signal transmitter and co-cultured with human bronchial epithelial cells BEAS-2B. Results showed that U937 cells were only activated by γ-irradiated NCI-H446 cells so that the secondary injuries in BEAS-2B cells under carbon ion irradiation were weaker than γ-rays. Both TNF-α and IL-1α were involved in the γ-irradiation induced secondary bystander effect but only TNF-α contributed to the carbon ion induced response. Further assay disclosed that IL-1α but not TNF-α was largely responsible for the activation of macrophages and the formation of micronucleus in BEAS-2B cells. These data suggest that macrophages could transfer secondary bystander signals and play a key role in the secondary bystander effect of photon irradiation, while carbon ion irradiation has conspicuous advantage due to its reduced secondary injury.

Target irradiation induced bystander effects between stem-like and non stem-like cancer cells.

Tumors are heterogeneous in nature and consist of multiple cell types. Among them, cancer stem-like cells (CSCs) are suggested to be the principal cause of tumor metastasis, resistance and recurrence. Therefore, understanding the behavior of CSCs in direct and indirect irradiations is crucial for clinical radiotherapy. Here, the CSCs and their counterpart non stem-like cancer cells (NSCCs) in human HT1080 fibrosarcoma cell line were sorted and labeled, then the two cell subtypes were mixed together and chosen separately to be irradiated via a proton microbeam. The radiation-induced bystander effect (RIBE) between the CSCs and NSCCs was measured by imaging 53BP1 foci, a widely used indicator for DNA double strand break (DSB). CSCs were found to be less active than NSCCs in both the generation and the response of bystander signals. Moreover, the nitric oxide (NO) scavenger c-PTIO can effectively alleviate the bystander effect in bystander NSCCs but not in bystander CSCs, indicating a difference of the two cell subtypes in NO signal response. To our knowledge, this is the first report shedding light on the RIBE between CSCs and NSCCs, which might contribute to a further understanding of the out-of-field effect in cancer radiotherapy.

Radioactive contamination mapping of northeastern and eastern Japan by a car-borne survey system, Radi-Probe.

We constructed a new car-borne survey system called Radi-Probe with a portable germanium gamma-ray spectrometer onboard a cargo truck, to identify radionuclides and quantify surface contamination from the accident at Fukushima Dai-ichi Nuclear Power Station. The system can quickly survey a large area and obtain ambient dose equivalent rates and gamma-ray energy spectra with good energy resolution. We also developed a new calibration method for the system to deal with an actual nuclear disaster, and quantitative surface deposition densities of radionuclides, such as (134)Cs and (137)Cs, and kerma rates of each radionuclide can be calculated. We carried out car-borne survey over northeastern and eastern Japan (Tohoku and Kanto regions of Honshu) from 25 September through 7 October 2012. We discuss results of the distribution of ambient dose equivalent rate H(∗)(10), (134)Cs and (137)Cs surface deposition densities, spatial variation of (134)Cs/(137)Cs ratio, and the relationship between surface deposition densities of (134)Cs/(137)Cs and H(∗)(10). The ratio of (134)Cs/(137)Cs was nearly constant within our measurement precision, with average 1.06 ± 0.04 in northeastern and eastern Japan (decay-corrected to 11 March, 2011), although small variations from the average were observed.

Co-visualization of DNA damage and ion traversals in live mammalian cells using a fluorescent nuclear track detector.

The geometric locations of ion traversals in mammalian cells constitute important information in the study of heavy ion-induced biological effect. Single ion traversal through a cellular nucleus produces complex and massive DNA damage at a nanometer level, leading to cell inactivation, mutations and transformation. We present a novel approach that uses a fluorescent nuclear track detector (FNTD) for the simultaneous detection of the geometrical images of ion traversals and DNA damage in single cells using confocal microscopy. HT1080 or HT1080-53BP1-GFP cells were cultured on the surface of a FNTD and exposed to 5.1-MeV/n neon ions. The positions of the ion traversals were obtained as fluorescent images of a FNTD. Localized DNA damage in cells was identified as fluorescent spots of γ-H2AX or 53BP1-GFP. These track images and images of damaged DNA were obtained in a short time using a confocal laser scanning microscope. The geometrical distribution of DNA damage indicated by fluorescent γ-H2AX spots in fixed cells or fluorescent 53BP1-GFP spots in living cells was found to correlate well with the distribution of the ion traversals. This method will be useful for evaluating the number of ion hits on individual cells, not only for micro-beam but also for random-beam experiments.

Measurement of air dose rates over a wide area around the Fukushima Dai-ichi Nuclear Power Plant through a series of car-borne surveys.

A series of car-borne surveys using the Kyoto University RAdiation MApping (KURAMA) and KURAMA-II survey systems has been conducted over a wide area in eastern Japan since June 2011 to evaluate the distribution of air dose rates around the Fukushima Dai-ichi Nuclear Power Plant and to evaluate the time-dependent trend of decrease in air dose rates. An automated data processing system for the KURAMA-II system was established, which enabled rapid analysis of large amounts of data obtained using about 100 KURAMA-II units. The initial data used for evaluating the migration status of radioactive cesium were obtained in the first survey, followed by other car-borne surveys conducted over more extensive and wider measurement ranges. By comparing the measured air dose rates obtained in each survey (until December 2012), the decreasing trend of air dose rates measured through car-borne surveys was found to be more pronounced than those expected on the basis of the physical decay of radioactive cesium and of the air dose rates measured using NaI (Tl) survey meters in the areas surrounding the roadways. In addition, it was found that the extent of decrease in air dose rates depended on land use, wherein it decreased faster for land used as building sites than for forested areas.

Differential effects of p53 on bystander phenotypes induced by gamma ray and high LET heavy ion radiation.

High LET particle irradiation has several potential advantages over γ-rays such as p53-independent response. The purpose of this work is to disclose the effect of p53 on the bystander effect induced by different LET irradiations and underlying mechanism. Lymphocyte cells of TK6 (wild type p53) and HMy2.CIR (mutated p53) were exposed to either low or high LET irradiation, then their mitochondrial dysfunction and ROS generation were detected. The micronuclei (MN) induction in HL-7702 hepatocytes co-cultured with irradiated lymphocytes was also measured. It was found that the mitochondrial dysfunction, p66(Shc) activation, and intracellular ROS were enhanced in TK6 but not in HMy2.CIR cells after γ-ray irradiation, but all of them were increased in both cell lines after carbon and iron irradiation. Consistently, the bystander effect of MN formation in HL-7702 cells was only triggered by γ-irradiated TK6 cells but not by γ-irradiated HMy2.CIR cells. But this bystander effect was induced by both lymphocyte cell lines after heavy ion irradiation. PFT-µ, an inhibitor of p53, only partly inhibited ROS generation and bystander effect induced by 30 keV/µm carbon-irradiated TK6 cells but failed to suppress the bystander effect induced by the TK6 cells irradiated with either 70 keV/µm carbon or 180 keV/µm iron. The mitochondrial inhibitors of rotenone and oligomycin eliminated heavy ion induced ROS generation in TK6 and HMy2.CIR cells and hence diminished the bystander effect on HL-7702 cells. These results clearly demonstrate that the bystander effect is p53-dependent for low LET irradiation, but it is p53-independent for high LET irradiation which may be because of p53-independent ROS generation due to mitochondrial dysfunction.

Gap junction communication and the propagation of bystander effects induced by microbeam irradiation in human fibroblast cultures: the impact of radiation quality.

Understanding the mechanisms underlying the bystander effects of low doses/low fluences of low- or high-linear energy transfer (LET) radiation is relevant to radiotherapy and radiation protection. Here, we investigated the role of gap-junction intercellular communication (GJIC) in the propagation of stressful effects in confluent normal human fibroblast cultures wherein only 0.036-0.144% of cells in the population were traversed by primary radiation tracks. Confluent cells were exposed to graded doses from monochromatic 5.35 keV X ray (LET ~6 keV/µm), 18.3 MeV/u carbon ion (LET ~103 keV/µm), 13 MeV/u neon ion (LET ~380 keV/µm) or 11.5 MeV/u argon ion (LET ~1,260 keV/µm) microbeams in the presence or absence of 18-α-glycyrrhetinic acid (AGA), an inhibitor of GJIC. After 4 h incubation at 37°C, the cells were subcultured and assayed for micronucleus (MN) formation. Micronuclei were induced in a greater fraction of cells than expected based on the fraction of cells targeted by primary radiation, and the effect occurred in a dose-dependent manner with any of the radiation sources. Interestingly, MN formation for the heavy-ion microbeam irradiation in the absence of AGA was higher than in its presence at high mean absorbed doses. In contrast, there were no significant differences in cell cultures exposed to X-ray microbeam irradiation in presence or absence of AGA. This showed that the inhibition of GJIC depressed the enhancement of MN formation in bystander cells from cultures exposed to high-LET radiation but not low-LET radiation. Bystander cells recipient of growth medium harvested from 5.35 keV X-irradiated cultures experienced stress manifested in the form of excess micronucleus formation. Together, the results support the involvement of both junctional communication and secreted factor(s) in the propagation of radiation-induced stress to bystander cells. They highlight the important role of radiation quality and dose in the observed effects.