Late effects from particulate radiations in primate and rabbit tissues.

Optic tissues in groups of New Zealand white rabbits were irradiated locally at different stages throughout the median life span of the species with a single dose (9 Gy) of 425 MeV/amu Ne ions (LET infinity approximately 30 keV/micrometer) and then inspected routinely for the progression of radiation cataracts. The level of early cataracts was found to be highest in the youngest group of animals irradiated (8 weeks old), but both the onset of late cataracts and loss of vision occurred earlier when animals were irradiated during the second half of the median life span. This age response can have serious implications in terms of space radiation hazards to man. Rhesus monkeys that had been subjected to whole-body skin irradiation (2.8 and 5.6 Gy) by 32 MeV protons (range in tissue approximately 1 cm) some twenty years previously were analysed for radiation damage by the propagation of skin fibroblasts in primary cultures. Such propagation from skin biopsies in MEM-alpha medium (serial cultivation) or in supplemented Ham's F-10 medium (cultivation without dilution) revealed late damage in the stem (precursor) cells of the skins of the animals. The proton fluxes employed in this experiment are representative of those occurring in major solar flares.     

Late skin damage in rabbits and monkeys after exposure to particulate radiations.

Skin biopsies were taken from the central regions of the ears of New Zealand white rabbits following localized exposure of one ear of each rabbit to 530 MeV/amu Ar or 365 MeV/amu Ne ions. The unirradiated ears served as controls. Biopsies were taken also from the chests and inner thighs of rhesus monkeys after whole-body exposure to 32 MeV protons and from unirradiated control animals. The linear energy transfers (LET infinity's) for the radiations were 90 +/- 5, 35 +/- 3, and approximately 1.2 keV/micrometer, respectively. In the rabbit studies, explants were removed with a 2 mm diameter dermal punch at post-irradiation times up to five years after exposure. Similar volumes of monkey tissue were taken from skin samples excised surgically 16-18 years following proton irradiation. Fibroblast cultures were initiated from the explants and were propagated in vitro until terminal senescence (cessation of cell division) occurred. Cultures from irradiated tissue exhibited decreases in doubling potential that were dependent on radiation dose and LET infinity and seemed to reflect damage to stem cell populations. The implications of these results for astronauts exposed to heavy ions and/or protons in space include possible manifestations of residual effects in the skin many years after exposure (e.g. unsatisfactory responses to trauma or surgery).     

Chromosome aberrations in human lymphocytes induced by 250 MeV protons: effects of dose, dose rate and shielding.

Although the space radiation environment consists predominantly of energetic protons, astronauts inside a spacecraft are chronically exposed to both primary particles as well as secondary particles that are generated when the primary particles penetrate the spacecraft shielding. Secondary neutrons and secondary charged particles can have an LET value that is greater than the primary protons and, therefore, produce a higher relative biological effectiveness (RBE). Using the accelerator facility at Loma Linda University, we exposed human lymphocytes in vitro to 250 MeV protons with doses ranging from 0 to 60 cGy at three different dose rates: a low dose rate of 7.5 cGy/h, an intermediate dose rate of 30 cGy/h and a high dose rate of 70 cGy/min. The effect of 15 g/cm2 aluminum shielding on the induction of chromosome aberrations was investigated for each dose rate. After exposure, lymphocytes were incubated in growth medium containing phytohemagglutinin (PHA) and chromosome spreads were collected using a chemical-induced premature chromosome condensation (PCC) technique. Aberrations were analyzed using the fluorescence in situ hybridization (FISH) technique with three different colored chromosome-painting probes. The frequency of reciprocal and complex-type chromosome exchanges were compared in shielded and unshielded samples.     

Induction of high grade astrocytoma (HGA) by protons: molecular mechanisms and RBE considerations.

Protons of a specific energy, 55 MeV, have been found to induce primary high grade astrocytomas (HGA) in the Rhesus monkey (Macaca mulatta). Brain tumors of this type were not induced by protons of other energies (32-2,300 MeV). Induction of HGA has been identified in human patients who have had radiation therapy to the head. We believe that the induction of HGA in the monkey is a consequence of dose distribution, not some unique "toxic" property of protons. Comparison of the human experience with the monkey data indicates the RBE for induction of brain tumors to be about one. It is unlikely that protons cause an unusual change in oncogenic expression, as compared to conventional electromagnetic radiation.     

Trapped antiprotons produced by cosmic rays in the Earth's magnetosphere.

The existence of significant fluxes of antiparticles in the Earth magnetosphere has been predicted on theoretical considerations in this article. These antiparticles (positrons or antiprotons) at several hundred kilometers of altitudes, we believe are not of direct extraterrestrial origin, but are the natural products of nuclear reactions of the high energy primary cosmic rays (CR) and trapped protons (TP) confined in the terrestrial radiation belt, with the constituents of terrestrial atmosphere. Extraterrestrial positrons and antiprotons born in nuclear reactions of the same CR particles passing through only 5-7 g/cm2 of interstellar matter, exhibit lower fluxes compared to the antiprotons born at hundreds of g/cm2 in the atmosphere, which when confined in the magnetic field of the Earth (in any other planet), get accumulated. We present the results of the computations of the antiproton fluxes at 10 MeV to several GeV energies due to CR particle interactions with the matter in the interstellar space, and also with the residual atmosphere at altitudes of approximately 1000 km over the Earth's surface. The estimates show that the magnetospheric antiproton fluxes are greater by two orders of magnitude compared to the extraterrestrial fluxes measured at energies <1-2 GeV.     

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.     

Protein expression profile changes in human fibroblasts induced by low dose energetic protons

Extrapolation of known radiation risks to the risks from low dose and low dose-rate exposures of human population, especially prolonged exposures of astronauts in the space radiation environment, relies in part on the mechanistic understanding of radiation induced biological consequences at the molecular level. While some genomic data at the mRNA level are available for cells or animals exposed to radiation, the data at the protein level are still lacking. Here, we studied protein expression profile changes using Panorama antibody microarray chips that contain antibodies to 224 proteins (or their phosphorylated forms) involved in cell signaling that included mostly apoptosis, cytoskeleton, cell cycle and signal transduction. Normal human fibroblasts were cultured until fully confluent and then exposed to 2 cGy of 150 MeV protons at high-dose rate. The proteins were isolated at 2 or 6 h after exposure and labeled with Cy3 for the irradiated cells and with Cy5 for the control samples before loading onto the protein microarray chips. The intensities of the protein spots were analyzed using ScanAlyze software and normalized by the summed fluorescence intensities and the housekeeping proteins. The results showed that low dose protons altered the expression of more than 10% of the proteins listed in the microarray analysis in various protein functional groups. Cell cycle (24%) related proteins were induced by protons and most of them were regulators of G1/S-transition phase. Comparison of the overall protein expression profiles, cell cycle related proteins, cytoskeleton and signal transduction protein groups showed significantly more changes induced by protons compared with other protein functional groups.     

The heliolongitudinal distribution of solar flares associated with solar proton events.

We find that the heliolongitudinal distribution of solar flares associated with earth-observed solar proton events is a function of the particle measurement energy. For solar proton events containing fluxes with energies exceeding 1 GeV, we find a Gaussian distribution about the probable root of the Archimedean spiral favorable propagation path leading from the earth to the sun. This distribution is modified as the detection threshold is lowered. For > 100 MeV solar proton events with fluxes > or = 10 protons (cm2-sec-ster)-1 we find the distribution becomes wider with a secondary peak near the solar central meridian. When the threshold is lowered to 10 MeV the distribution further evolves. For > 10 MeV solar proton events having a flux threshold at 10 protons (cm2-sec-ster)-1 the distribution can be considered to be a composite of two Gaussians. One distribution is centered about the probable root of the Archimedean spiral favorable propagation path leading from the earth to the sun, and the other is centered about the solar central meridian. For large flux solar proton events, those with flux threshold of 1000 (cm2-sec-ster)-1 at energies > 10 MeV, we find the distribution is rather flat for about 40 degrees either side of central meridian.     

Relative biological effectiveness and microdosimetry of a mixed energy field of protons up to 200 MeV.

We have studied radiation effects utilizing the new 250 MeV Synchrotron at Loma Linda University Medical Center. In this paper we present the data collected for the survival of Chinese hamster lung (V79) cells, that were irradiated with a beam of mixed energy protons up to 200 MeV. The RBE for protons, when compared to 60Co gamma rays, ranged from a low of 1.2 at the high energy portion of the field to 1.3+ at the low energy portion of the field. These results are consistent with the measured lineal energy (microdosimetric) spectra.     

Monte Carlo calibration of the response of the University of Chicago’s Cosmic Ray Nuclei Experiment (CRNE) on IMP-8 to electrons above 0.5 MeV

Modern instrument-simulation techniques offer the possibility of increasing the scientific yield from archival space datasets. In this paper, we report on a simulation of the electron response of the University of Chicago’s Cosmic Ray Nuclei Experiment (CRNE) instrument on the IMP-8 satellite. IMP-8/CRNE returned data from 1973 to 2006. The CRNE particle telescope was designed to measure the isotopic composition of Galactic cosmic-ray (GCR) nuclei and has also been used in many studies of protons and ions above 10 MeV/nucleon from solar energetic particle (SEP) events. But CRNE also functions as a highly-capable detector for solar electrons above 0.5 MeV, an energy range that has not been extensively studied. Utilization of the CRNE electron data has heretofore been limited by the fact that CRNE was never calibrated for electrons. We have therefore used the GEANT4 Monte Carlo simulation package to model the CRNE response to electrons and (separately) protons for multiple energies and incident angles. The results were used to compute the energy- and angle-dependence of the effective area and the energy-dependence of the geometric factor. The response to protons, which was already well understood, was used to verify the mass model, the simulation settings, and the post-processing software. Our simulation of the IMP-8/CRNE electron response now allows analysis of hundreds of relativistic solar electron events observed by CRNE over the years, including studies of evolution of electron energy spectra with high time resolution. We show examples of these results and briefly discuss potential applications to future scientific investigations.     

A model of radiation conditions during spacecraft flights in the interplanetary space and in the Earth's magnetosphere.

Based on the available measurement data, simulations of radiation conditions during spacecraft flights in the interplanetary space and in the Earth's and Jupiter's radiation belts has been carried out. The > or = 10 MeV and > or = 30 MeV solar flare proton fluence forecast has been proposed for Cycle 22. Radiation conditions due to both magnetospheric electrons and protons and to solar flare protons, magnetic rigidity cutoff being taken into account, have been evaluated on spacecraft trajectories in the Earth's and Jupiter's magnetospheres.     

Late cataractogenesis in primates and lagomorphs after exposure to particulate radiations.

Rhesus monkeys that were exposed in 1969, at the age of approximately 2 years, to low doses of "mixed-energy" protons (10- and 110-MeV) are exhibiting progressive (degenerative) lenticular changes. We have conducted regular examinations of this group of monkeys for cataractogenic development since 1987, i.e., 18 years after irradiation, and the animals began to show enhanced degrees of lenticular opacification two years later. The lenses of age-matched controls (median lifespan in captivity approximately 24 years) continue to exhibit much lower levels of opacification (senile cataracts). Trends in the new data are consistent with the cataractogenic patterns observed for other groups of monkeys that were exposed at similar ages in 1964 and 1965 to protons of different energies, and which we began to monitor only 20-21 years later. Therefore, the new information from the mixed-energy group of monkeys provides insight into the development of late cataractogenic sequelae in the other groups of animals during the 2-3 years before we began to measure them. Comparisons are also made here among recent results from the different groups of primates and from New Zealand white (NZW) rabbits that were exposed when young to 56Fe ions and monitored continuously thereafter. This is done because analogous expression of radiation-induced degenerative cataractogenesis also occurs late in the lifespan of the lagomorphs (control median lifespan in captivity approximately 5-7 years), but in this case the cataractogenic profile has been documented through most of the post-irradiation lifespan.     

Single ion actions: the induction of micronuclei in V79 cells exposed to individual protons.

Understanding the effects of single-particles from conventional radiation biology experiments is problematic due to the stochastics of particle tracks. This complicates the determinations of risk associated with low doses. We have developed a charged particle microbeam, which allows individually counted particles to be delivered to precise cellular locations. The system is capable of delivering a single charged particle with > 99% efficiency. Of these particles 90% are delivered with a resolution of +/- 2 micrometers and 96% with a resolution of +/- 5 micrometers. We have carried out preliminary studies in Chinese hamster V79 cells to monitor the effectiveness of low energy protons at inducing cytological damage. We have used the micronucleus assay as a measure of predominantly lethal chromosome damage. The effects of a single 3.2 MeV proton delivered individually to cells could be measured, with less than 2% of the exposed cells producing micronuclei 24 hours later. The yield of micronuclei formation was essentially linear up to the highest dose (30 particles per cell nucleus) delivered. Ultimately, the ability to target particles to different parts of the cell nucleus may start to impact on models available for chromosome aberration formation and chromosomal Organisation and mechanisms underlying genomic instability.     

The effects of proton exposure on neurochemistry and behavior.

Future space missions will involve long-term travel beyond the magnetic field of the Earth, where astronauts will be exposed to radiation hazards such as those that arise from galactic cosmic rays. Galactic cosmic rays are composed of protons, alpha particles, and particles of high energy and charge (HZE particles). Research by our group has shown that exposure to HZE particles, primarily 600 MeV/n and 1 GeV/n 56Fe, can produce significant alterations in brain neurochemistry and behavior. However, given that protons can make up a significant portion of the radiation spectrum, it is important to study their effects on neural functioning and on related performance. Therefore, these studies examined the effects of exposure to proton irradiation on neurochemical and behavioral endpoints, including dopaminergic functioning, amphetamine-induced conditioned taste aversion learning, and spatial learning and memory as measured by the Morris water maze. Male Sprague-Dawley rats received a dose of 0, 1.5, 3.0 or 4.0 Gy of 250 MeV protons at Loma Linda University and were tested in the different behavioral tests at various times following exposure. Results showed that there was no effect of proton irradiation at any dose on any of the endpoints measured. Therefore, there is a contrast between the insignificant effects of high dose proton exposure and the dramatic effectiveness of low dose (<0.1 Gy) exposures to 56Fe particles on both neurochemical and behavioral endpoints.     

The quantification of wound healing as a method to assess late radiation damage in primate skin exposed to high-energy protons.

In an experiment examining the effects of space radiations on primates, different groups of rhesus monkeys (Macaca mulatta) were exposed to single whole-body doses of 32- or 55-MeV protons. Survivors of those exposures, together with age-matched controls, have been monitored continuously since 1964 and 1965. Late effects of nominal proton doses ranging from 2-6 Gray have been measured in vitro using skin fibroblasts from the animals. A logical extension of that study is reported here, and it involves observations of wound healing after 3-mm diameter dermal punches were removed from the ears (pinnae) of control and irradiated monkeys. Tendencies in the reduction of competence to repair cutaneous wounds have been revealed by the initial examinations of animals that received doses greater than 2 Gy more than 2 decades earlier. These trends indicate that this method of assessing radiation damage to skin exposed to high-energy radiations warrants further study.     

Neoplastic transformation of mouse C3H 10T1/2 and Syrian hamster embryo cells by heavy ions.

C3H 10T1/2 mouse-embryo fibroblasts were used for transformation experiments to study the effectiveness of various heavy ions with energies up to 20 MeV/u and LET values from 170 to 16,000 keV/micrometers. The transformation frequency per unit absorbed dose decreased with increasing ionization density; at the highest values of LET we found a decrease even of the transformation efficiency per unit fluence. Uranium ions at energies of 5, 9, and 16.3 MeV/u did not induce any transformation. In additional studies primary Syrian hamster embryo cells (SHE) were exposed to heavy ions in order to characterize cytological and molecular changes which may be correlated with neoplastic transformation. Growth behaviour, chromosomal status, tumorigenicity in nude mice, and expression of oncogenes of transformed cell lines were examined     

First-order Fermi shock acceleration in solar flares

First order Fermi shock acceleration of electrons, protons and alpha particles is compared to observations of energetic particle events. For each event, a unique shock compression ratio produces spectra in good agreement with observation. The simple model predicts that the acceleration time to a given energy will be approximately equal for electrons and protons and, for reasonable solar parameters, can be less than 1 second to ～ 100 MeV.     

Cataractogenic potential of ionizing radiations in animal models that simulate man.

Aspects of experiments on radiation-induced lenticular opacification during the life spans of two animal models, the New Zealand white rabbit and the rhesus monkey, are compared and contrasted with published results from a life span study of another animal model, the beagle dog, and the most recent data from the ongoing study of the survivors from radiation exposure at Hiroshima and Nagasaki. An important connection among the three animal studies is that all the measurements of cataract indices were made by one of the authors (A.C.L.), so variation form personal subjectivity was reduced to a minimum. The primary objective of the rabbit experiments (radiations involved: 56Fe, 40Ar and 20Ne ions and 60Co gamma photons) is an evaluation of hazards to astronauts from galactic particulate radiations. An analogous evaluation of hazards from solar flares during space flight is being made with monkeys exposed to 32, 55, 138 and 400 MeV protons. Conclusions are drawn about the proper use of animal models to simulate radiation responses in man and the levels of radiation-induced lenticular opacification that pose risks to man in space.     

Temporal variation in the new proton belt created in March 1991 observed using the CREAM & CREDO experiments.

The Cosmic Radiation Environment & Activation Monitor (CREAM) was carried in high inclination (57.1 degrees) orbits on Shuttle missions STS-48 in September 1991 (altitude 570 km) and STS-53 (altitude 325 to 385 km) in December 1992. On both occasions the instrument observed an excess of counts due to protons of greater than 30 MeV in energy in the region off of South Africa where field lines of L=2.5 intersect low earth orbit. Meanwhile the Cosmic Radiation Environment and Dosimetry Experiment (CREDO) carried to 840 km, 98.7 degrees orbit on UOSAT-3 has continued to sample the high field portions of the L-shells around L = 2.5 from April 1990 until the present time. When careful subtraction of cosmic-ray contributions is made it can be seen that the March 91 enhancement persisted for approximately 8 months and explains the STS-48 observation. There would appear to have been a further increase produced by the 31 October 1992 flare event and seen by STS-53.     

Distribution of energetic particles and secondary radiation according to orbital station “MIR” data obtained in 1991

A set of instruments for measuring energetic particle fluxes, containing two neutron detectors under different plexiglas shielding thicknesses, a scintillation detector, measuring energy release >0. I MeV and 0.5 MeV and a Geiger counter were launched onboard OS ‘MIR’. The latitude dependencies of the cosmic ray measurements were obtained and studied. The distributions of primary particle fluxes (protons and electrons) as well as secondary particle fluxes (bremsstrahlung gamma-rays and neutrons) produced in interactions of radiation belt particles with the station materials were obtained. The electron belt, generated during the storm of March 24 1991, is studied.