T. Straume

Impaired cell proliferation in mice that persists across at least two generations after paternal irradiation

Irradiation of male F0 mice 6 to 7 weeks prior to mating causes significant changes in the proliferation of F1 and F2 embryonic cells. These changes are revealed as a competitive cell proliferation disadvantage in chimera assays when the affected embryo is paired with a normal embryo in an aggregation chimera. This effect has been observed previously to be transmitted to F1 embryos for absorbed doses from 0.01 to 1.0 Gy; 0.01 Gy is about 100-fold lower than detectable using conventional germline mutation assays. However, until now there has been no reported cross-generation heritability. We now report that this competitive cell proliferation disadvantage persists without degradation in the F2 generation of embryos when F0 males received 1.0 Gy from gamma irradiation 6 and 7 weeks prior to conception of F1 males.

Neutrons confirmed in Nagasaki and at the Army pulsed radiation Facility: Implications for Hiroshima

Recent reports have clearly demonstrated that large discrepancies exist between neutron activation measured in Hiroshima and activation calculated using the current dosimetry system, DS86. The reports confirmed previous results for cobalt activation in Hiroshima that suggested problems, and this has spurred a joint U.S.-Japan effort to identify the source(s) of this discrepancy. Here, new results are presented that appear to eliminate both the measurements of neutron activation and the DS86 air-transport calculations as potential sources of the discrepancy in Hiroshima. Computer transport of DS86 fission neutrons through large distances of air was validated using concrete samples from Nagasaki and chloride detectors placed at selected distances from a bare uranium reactor. In both cases, accelerator mass spectrometry was used to measure thermal neutron activation via the reaction, 35Cl(n, gamma)36Cl (half-life, 301,000 years). Good agreement was observed between measurements of neutron activation and DS86 calculations for Nagasaki, as well as for the reactor experiment. Thus the large discrepancy observed in Hiroshima appears not to be due to uncertainties in air-transport calculations or in the activation measurements; rather, the discrepancy appears to be due to uncertainties associated with the Hiroshima bomb itself.

A search for neutron reactions that may be useful for Hiroshima dose reconstruction

Abstract Results are presented of a detailed search for neutron reactions that could potentially be useful in reconstructing the neutron fluence in Hiroshima, with emphasis on fast neutrons in the 1-MeV range. Searches were made for suitable reactions in several neutron cross-section libraries from the U.S., Europe, Japan, China, and the former U.S.S.R. Because of the long time (∼ 50 yr) since the atomic bombing of Hiroshima, reaction products evaluated in this search were limited to those with half-lives between 5 and 10 9 yr. From the about 100 reactions within this category, only six appear to have some potential utility for fast neutron measurements of suitable materials from Hiroshima. The technology may currently be available to measure two of these reactions if suitable materials can be obtained.

Measurement of neutron-induced genetic damage in mouse immature oocytes

Recent experimental evidence concerning the nature of radiosensitive targets in mouse immature (resting) oocytes has led to new experimental designs that permit measurement of radiation-induced genetic damage in these important cells. We have previously reported initial results of the detection of genetic damage in mouse immature oocytes using monoenergetic 0.43-MeV neutrons. Here we provide a full report of our data and compare the genetic sensitivity of immature oocytes with those measured by others for maturing oocytes. Until recently, all attempts to detect radiation-induced genetic damage in mouse immature oocytes had failed. This appears to have been because the radiation types and modes of dose delivery used in those studies did not sufficiently spare the hypersensitive lethality target (the plasma membrane) while at the same time deposit enough dose in DNA to produce detectable mutation. Recoil protons from 0.43-MeV neutrons produce short ionization tracks (2.6 micron mean) and can therefore deposit energy in the DNA without simultaneously traversing the plasma membrane. Using these particles, we have obtained dose-response relationships for both chromosome aberrations and dominant lethal mutations in oocytes from females irradiated 8-12 weeks earlier, when oocytes were immature. Results suggest that the intrinsic mutational sensitivity of mouse immature oocytes is not very different from that of maturing oocytes.

Neutron RBEs and the Radiosensitive Target for Mouse Immature Oocyte Killing

The highly radiosensitive immature oocytes of mice were irradiated in vivo with graded doses of 252Cf fission radiation, 0.43- or 15-MeV neutrons, or 60Co gamma rays. Comparisons of oocyte survival for neutrons and for gamma rays demonstrate that neutron RBEs for the killing of these important cells do not reach the high values (30-50 or more) at low doses observed for several other biological end points. Rather, neutrons differ little in effectiveness from gamma rays in killing these extremely sensitive murine oocytes. For 0.43-MeV neutrons, RBEs obtained from fitted survival curves reach only 1.7 at 0.1 rad. For 15-MeV neutrons, they are not significantly different from 1 at any dose tested (lowest, 4.5 rad). For 252Cf fission neutrons (E = 2.15 MeV), RBEs are intermediate between those for 0.43- and 15-MeV neutrons. For all neutron energies tested, the RBEs are particularly low in the juvenile period, a time when murine immature oocytes are especially radiosensitive. With exposure just prior to birth, however, when these cells are much less easily killed, higher, more usual RBEs are found. The minimum size of the lethality target in mouse immature oocytes, estimated from the inactivation constant for 0.43-MeV neutrons and microdosimetric values, is larger than the nucleus but not larger than the cell. This and related analytical considerations suggest that the hypersensitive target in these particular oocytes is the plasma membrane, a finding which is in excellent accord with results from other experiments using different, contrasting radiations and dose deliveries (accelerated Si14+ ions, gamma rays, and beta rays from 3HOH compared with those from [3H]thymidine).

Inherited effects from irradiated mouse immature oocytes detected in aggregation embryo chimeras

Data obtained using the mouse-preimplantation-embryo-chimera assay are presented that show a transmitted effect following low-dose irradiation of immature oocytes in vivo. Six-week-old female mice were irradiated using 137Cs-gamma-rays (0.05 Gy, 0.15 Gy, and unexposed controls). At 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 weeks post exposure, the mice were mated and aggregation chimeras made from the 4-cell embryos. Three independent experiments have now been carried out, all showing a significant embryonic cell-proliferation disadvantage of the embryos obtained from the females treated 7 weeks previously, i.e., embryos from oocytes that were immature at the time of radiation exposure. No effect was detected at 1-6 weeks when embryos were obtained from maturing oocytes. Also, the effect was not seen at 8, 9, 10, 11, and 12 weeks post exposure. The implications of these results are discussed in the light of previous studies on mouse oocytes.

Transmitted proliferation disadvantage from mouse oocytes labeled in vivo with [3H]thymidine: radiosensitive target considerations

This study was conducted to test the hypothesis that a nuclear target is involved in the embryonic cell proliferation disadvantage transmitted by irradiated mouse oocytes and detected by the chimera assay. In this assay, the cells from the irradiated embryo exhibit a competitive cell proliferation disadvantage when they are challenged by direct cell-cell contact with cells from a normal embryo in an aggregation chimera. Here, six pregnant CD-1 mice received a total of 1.85 TBq tritiated thymidine (TdR) delivered by multiple intraperitoneal injections during days 13-15 postconception. Six more pregnant mice were sham-injected to provide control embryos. Sixty randomly selected female progeny were mated at 47 days of age and their 4-cell embryos tested in the chimera assay. The mean proliferation ratio (PR, number of cells from the experimental embryo divided by total cell number of the chimera) for experimental chimeras was 0.45 +/- 0.02 SE (n = 43), which was significantly less than the mean PR of 0.49 +/- 0.01 SE (n = 47; p = 0.02) for control chimeras. This entire experiment was repeated, with similar results. A comparison for TdR confined to the nucleus (i.e., mean beta-ray range is only 0.7 microm) with the relationship for uniform irradiation by 137Cs gamma-rays demonstrates that these two very different modes of dose delivery produce essentially identical PRs. These results in vivo suggest a nuclear DNA target for embryonic cell proliferation disadvantage consistent with our previous findings in vitro.

Fast Neutrons Measured in Copper from the Hiroshima Atomic Bomb Dome

Abstract Marchetti, A. A., McAninch, J. E., Rugel, G., Rühm, W., Korschinek, G., Martinelli, R. E., Faestermann, T., Knie, K., Egbert, S. D., Wallner, A., Wallner, C., Tanaka, K., Endo, S., Hoshi, M., Shizuma, K., Fujita, S., Hasai, H., Imanaka, T. and Straume, T. Fast Neutrons Measured in Copper from the Hiroshima Atomic Bomb Dome. Radiat. Res. 171, 118–122 (2009). The first measurements of 63Ni produced by A-bomb fast neutrons (above ∼1 MeV) in copper samples from Hiroshima encompassed distances from ∼380 to 5062 m from the hypocenter (the point on the ground directly under the bomb). They included the region of interest to survivor studies (∼900 to 1500 m) and provided the first direct validation of fast neutrons in that range. However, a significant measurement gap remained between the hypocenter and 380 m. Measurements close to the hypocenter are important as a high-value anchor for the slope of the curve for neutron activation as a function of distance. Here we report measurements of 63Ni in copper samples from the historic Hiroshima Atomic Bomb Dome, which is located ∼150 m from the hypocenter. These measurements extend the range of our previously published data for 63Ni providing a more comprehensive and consistent A-bomb activation curve. The results are also in good agreement with calculations based on the current dosimetry system (DS02) and give further experimental support to the accuracy of this system that forms the basis for radiation risk estimates worldwide.

Mouse immature oocytes irradiated in vivo at 14-days of age and evaluated for transmitted effects using the aggregation embryo chimera assay

A previous study using the mouse-preimplantation-embryo-chimera assay demonstrated a reproducible transmitted effect (proliferation disadvantage observed in early embryos) from females irradiated as 49-day-old adults using 0.15 Gy of gamma rays and then mated seven weeks later, i.e., embryos were from oocytes that were immature at time of irradiation. Because mouse immature oocytes are known to be much more radiosensitive to cell killing in juveniles than in adults, a follow-on study was performed here using 14-day-old juvenile mice. In contrast to adults, the exposure of juveniles to 0.15 Gy of gamma rays did not result in a detectable transmitted proliferation disadvantage when animals were mated 7 or 12 weeks later. This observation is discussed in light of previous studies on mouse immature oocytes and embryo chimeras.