Reducing the radiation dose from inhaled americium-241 using continuously administered DTPA therapy

Accelerating the removal of a radionuclide from the body of a contaminated individual is the only available approach to decreasing the radiation dose from such exposures. In this study, continuous infusion of a chelating agent, DTPA, was given to dogs that had inhaled a moderately soluble aerosol, 241 AmO2, not only to accelerate clearance of the radionuclide from the lung but also to prevent its deposition in liver and bone. Treatment was begun with an intravenous injection of CaDTPA 1 h after exposure, and was continued for 64 days after exposure by implanting subcutaneously osmotic pumps containing ZnDTPA at 1 day after exposure. The results showed that the infusion therapy was effective in blocking the translocation of 99.5 per cent of the 241Am that would have been deposited in liver, and 98.3 per cent of the 241Am that would have been deposited in bone. This result was significantly better than the result achieved using repeated intravenous injections of DTPA, the method of treatment in current use for actinide contamination cases.     

Inhalation carcinogenesis of high-fired 241AmO2 in rats

Wistar rats were given a single inhalation exposure to high-fired 241AmO2 particles and examined over their life span. A total of 310 rats were used: 259 exposed to 241Am for life-span study, 30 exposed to 241Am for early metabolism study, and 21 unexposed life-span controls. The activity median aerodynamic diameter of the aerosols was 0.75-1.39 microns. About 55% of alveolarly deposited 241Am was cleared from the lung with a half-life of 0.5 days, 37% with a half-life of 7 days, and 8% with a half-life of 580 days. Group mean lung doses ranged from less than 5.7 rad up to 1500 rad. Significant early mortality due to radiation pneumonitis was seen only in the highest exposure group. The percentage of rats with lung tumors was 0% for controls (21 rats), 1% at lifetime lung doses less than 10 rad (139 rats), 7% at 10-50 rad (86 rats), 0% at 50-100 rad (9 rats), 60% at 100-500 rad (10 rats), and 7% at 500 rad (15 rats). Only one liver and one bone tumor were found in all exposed rats, both at lifetime tissue doses less than 10 rad. The fate and carcinogenicity of inhaled 241AmO2 in the lung of rats were similar to what has previously been described for inhaled 244CmO2.     

Retention and dosimetry of injected 241Am in beagles

Equations have been derived, from the results of total-body and partial-body counting and gamma-ray counting of individual bones and soft tissues, which describe the retention of injected 241Am in the liver, in the nonliver tissue (including skeleton), and in the skeleton of young adult beagles. Retention was found to be dependent upon injection level, and different sets of equations were developed for dogs given about (a) 2.8 microCi/kg (b) 0.9 microCi/kg (c) 0.3 microCi/kg, and (d) 0.1 microCi/kg and less. Liver rention, RL, was characterized by a single exponential equation of the form RL = ce-beta t, with c = 0.49 +/- 0.04 and beta = a function of injection level. Nonliver tissue was assigned a retention equation of the form RNL = d + alpha + J(l - e-mt), with d = 0.102 +/- 0.024 e-1.22t, alpha = 0.41 +/- 0.04, and both J and m as a function of injection level. Skeletal retention was found to be about 0.885 +/- 0.037 of nonliver retention with no significant dependence upon either injection level or time after 241Am injection. Dosimetry equations based on these retention expressions were derived. Individual bones of 55 beagles were assayed at death for their 241Am content for a determination of 241Am distribution within the skeleton.     

Assessment of the exposure pathway in the uptake and distribution of americium and cesium in cuttlefish (Sepia officinalis) at different stages of its life cycle

Laboratory radiotracer experiments were performed to study the uptake, assimilation and retention of americium (²⁴¹Am) and cesium (¹³⁴Cs) by the common cuttlefish Sepia officinalis. Uptake and loss kinetics of the radionuclides were measured following exposure through sediments, seawater and food at different stages of the animal's life cycle. Sediment was found to be a minor uptake pathway for both radionuclides in juveniles. Following a short seawater exposure, cuttlefish accumulated ²⁴¹Am and ¹³⁴Cs, but only to a limited extent (whole-body CF < 2). Among the cuttlefish organs, branchial hearts and their appendages displayed the highest degree of uptake for ²⁴¹Am (CF = 42 and 16, respectively), but these tissues contained low percentage of total ²⁴¹Am due to their relatively small contribution to whole organism weight. The major fraction of incorporated radionuclides was associated with muscular tissues (viz. 65% and 82% of total ²⁴¹Am and ¹³⁴Cs, respectively). Whole-body loss of ²⁴¹Am and ¹³⁴Cs was relatively rapid (T_b½ = 14 and 6 days, respectively). After dietary exposure, around 60% and 30% of ingested ²⁴¹Am was assimilated into the tissues of juvenile and adult cuttlefish, respectively. However, assimilated ²⁴¹Am was more strongly retained in adults than in juveniles (T_b½ = 28 vs. 5 days, respectively), suggesting that different mechanisms govern ²⁴¹Am elimination at both ages. Ingested ¹³⁴Cs was assimilated to a similar extent in juveniles (29%) and adults (23%), but the depuration rate was four times faster in adults. Our results strongly suggest that these two radionuclides follow different excretion pathways and that the mechanisms can vary with age for a given radionuclide.     

Effect of gavaged chemical form of 241Am on its retention in mice

The retention of 241Am in mice 48 h after administration by gavage is reported here. The 241Am was given to mice in the form of either 241Am nitrate or 241Am citrate. The 241Am was also injected into rats in the same form. The homogenized livers of those rats were subsequently administered by gavage to another group of mice. The retention of 241Am citrate was 1.5 X 10(-2)% of the original dose and was the highest among the compounds examined. The retention of biologically incorporated 241Am into the liver as 241Am nitrate and as 241Am citrate was 2.4 X 10(-3) and 2.6 X 10(-3)%, respectively, and was similar to the retention of 241Am nitrate, which was 2.8 X 10(-3)%. The ratio of the retention in the carcass to that in the liver for the 241Am citrate was lower than that of the 241Am nitrate and the biologically incorporated 241Am. This difference indicates that the distribution of 241Am in the animal body depends on the chemical form administered. The retention of liver-incorporated 241Am as citrate after autolysis of the liver is similar to that of fresh liver-incorporated 241Am citrate.     

Influence of radiation dose patterns on lung tumor incidence in dogs that inhaled beta emitters: a preliminary report

Different radiation dose patterns to the lung from inhaled beta-emitting radionuclides may influence the frequency and kind of biological effects. To determine the magnitude of this influence, groups of Beagle dogs were exposed to aerosols of 90Y, 91Y, 144Ce, or 90Sr in relatively insoluble particles and observed for their life spans. Different dose patterns were achieved by using these radionuclides having similar beta emissions and chemical form but having physical half-lives ranging from 2.6 days to 28 years. The range of initial lung burdens of radionuclides studied resulted in a range of biological effects from early deaths at the highest radiation doses to no discernible effects at the lowest doses. The effective half-lives of the four radionuclides in the lung ranged from 2.5 to 600 days. Within 1.5 years after exposure, some dogs died with radiation pneumonitis and pulmonary fibrosis. Between 1.5 and 10 years after exposure, 42 pulmonary carcinomas and 28 pulmonary sarcomas were observed in 163 dogs that died. Protracted irradiation of the lung from 90Sr or 144Ce resulted in a relatively high radiation dose and produced more total lung tumors but fewer lung tumors per rad than less protracted irradiation from 90Y or 91Y. At 10 years after inhalation exposure, the difference in risk per rad among the different dose patterns was a factor of 4 to 8, indicating that the different radiation dose patterns from inhaled beta emitters do influence lung tumor risk factors, at least at high (greater than 20,000 rad) doses to lung.     

Lung lavage therapy to lessen the biological effects of inhaled 144Ce in dogs

To evaluate the therapeutic effects of removal of an internally deposited radionuclide on long-term biological effects, lung lavage was used to treat dogs that had inhaled 144Ce in a relatively insoluble form, in fused aluminosilicate particles. Either 10 lung lavages were performed between Days 2 and 56 after exposure or 20 lung lavages were performed between Days 2 and 84 after exposure. Approximately one-half of the 144Ce was removed by the lavages, resulting in a corresponding reduction in the total absorbed beta dose to lung. The mean survival time of the treated dogs was 1270 days compared to 370 days for untreated dogs whose initial pulmonary burdens of 144Ce were similar. Treated dogs died late from cancers of the lung or liver, whereas the untreated dogs died at much earlier times from radiation pneumonitis. Dogs treated with lung lavage but not exposed to 144Ce had a mean survival of 4770 days. We concluded that removal of 144Ce from the lung by lavage resulted in increased survival time and in a change in the biological effects from inhaled 144Ce from early-occurring inflammatory disease to late-occurring effects, principally cancer. In addition, the biological effects occurring in the treated dogs could be better predicted from the total absorbed beta dose in the lung and the dose rate after treatment rather than from the original dose rate to the lung. Therefore, we concluded that prompt treatment to remove radioactive materials could be of significant benefit to persons accidentally exposed to high levels of airborne, relatively insoluble, radioactive particles.     

Effect of inhaled alpha-emitting nuclides on mouse alveolar macrophages

The effects of inhaled alpha emitters on the free cell population of the mouse lung were investigated up to 100 days after exposure. Groups of mice inhaled aerosols of 238PuO2, 239PuO2, or 241Am(NO3)3 to give alveolar deposits resulting in lung-averaged cumulative absorbed doses of about 20 Gy by the end of the study. Initially, with 238Pu most of the activity was associated with relatively few pulmonary alveolar macrophages (PAM), whereas with 241Am, all pulmonary alveolar macrophages were labeled and a substantial fraction was extracellular. The free cell population of the lung was sampled using bronchoalveolar lavage. The main parameters investigated were (a) the recovery and total numbers of free cells, including PAM, lymphocytes, and neutrophils; (b) the incidence of nuclear abnormalities in PAM (cells with more than one nucleus or with micronuclei); and (c) metabolic activation of PAM from measurements of their size and associated beta-glucuronidase activity. All three actinides produced depletions in total numbers of PAM, increased incidences of nuclear abnormalities, and metabolic activation of PAM, without a marked infiltration of inflammatory cells. Americium-241, which is distributed relatively uniformly in PAM, produced the most marked changes in that population and 238Pu, which gave the most inhomogeneous distribution of activity, produced the least.     

Removal of Pu and am from beagles and mice by 3,4,3-LICAM(C) or 3,4,3-LICAM(S)

Decorporation of Pu and Am by tetrameric catechoylamide (CAM) ligands has been investigated in beagles and mice. Eight dogs were injected intravenously (iv) with 237 + 239Pu(IV) + 241Am(III) citrate, and 30 min later, pairs of dogs were injected iv with 30 mumole/kg of 3,4,3-LICAM(C) [N1,N5,N10,N14-tetrakis(2,3-dihydroxy-5-sulfobenzoyl)tetr aazatetradecane, tetrasodium salt], 3,4,3-LICAM(S) [N1,N5,N10,N14-tetrakis(2,3-dihydroxy-4-carboxybenzoyl)te traazatetradecane, tetrasodium salt], CaNa3-DTPA, or each of the latter two ligands. Blood was sampled, and excreta were collected for 7 days, at which time the dogs were sacrificed and nuclide retention in liver and nonliver tissue was measured. Groups of five mice were each given 238Pu(IV) or 241Am(III) citrate iv; 3 min later 30 mumole/kg of a CAM ligand was injected intraperitoneally, mice were killed at 24 hr, and separated excreta and tissues were analyzed. In the dogs, average retention at 7 days of the injected Pu and Am, respectively, was as follows: 12 and 70% after treatment with a CAM ligand alone; 30 and 20% after DTPA; 12 and 20% after LICAM(S) plus DTPA; 90 and 89% without a ligand. In the mice, mean retention of the injected Pu and Am, respectively, was as follows: 14 and 66% after treatment with LICAM(C); 21 and 54% after LICAM(S); 91 and 87% without a ligand. In both species, about 99% of net Pu excretion (excretion with ligand - excretion without ligand) promoted in 24 hr by DTPA or LICAM(S) was in the urine, whereas about 10% of net Pu excretion promoted by the less hydrophilic LICAM(C) was in feces. Delayed excretion of both Am and Pu was significant in all ligand-treated dogs. Comparison of the nuclide content of tissues of ligand-treated mice with those of mice killed 3 min after nuclide injection indicated that the CAM ligands chelated circulating Pu and Am and prevented further deposition. In addition, the CAM ligands removed much of the presumably loosely bound Pu present in liver and skeleton at the time of ligand injection. LICAM(C) was more effective in removing Pu from liver and LICAM(S) was more effective in the skeleton. Moderate to severe uremia and histological evidence of cell killing in the distal tubules of the kidney were observed in the four dogs injected once with 30 mumole/kg of LICAM(S).(ABSTRACT TRUNCATED AT 400 WORDS)     

Effect of low-level alpha-radiation on bioluminescent assay systems of various complexity.

This study addresses the effects of low-level alpha-radiation on bioluminescent assay systems of different levels of organization: in vivo and in vitro. Three bioluminescent assay systems are used: intact bacteria, lyophilized bacteria, and bioluminescent system of coupled enzyme reactions. Solutions of 241Am(NO3)3 are used as a source of alpha-radiation. It has been shown that activation processes predominate in all the three bioluminescent assay systems subjected to short-term exposure (20-55 h) and inhibition processes in the systems subjected to longer-term exposure to radiation. It has been found that these effects are caused by the radiation component of 241Am3+ impact. The intensity of the 241Am3+ effect on the bioluminescent assay systems has been shown to depend on the 241Am3+ concentration, level of organization and integrity of the bioluminescent assay system. The bioluminescent assay systems in vivo have been found to be highly sensitive to 241Am3+ (up to 10(-17) M).     

Plutonium microdistribution in the lungs of Mayak workers

The degree of nonuniform distribution of plutonium in the human lung has not been determined; thus current dosimetric models do not account for nonuniform irradiation. A better scientific basis is needed for assessing the risk of developing radiation-induced disease from inhaled alpha-particle-emitting radionuclides. We measured the distribution of plutonium activity in the lung by autoradiography and related the activity to specific compartments of the lung. The study materials were lung specimens from deceased workers employed by the Mayak Production Association. The approach to analyzing these lung samples used contemporary stereological sampling and analysis techniques together with quantitative alpha-particle autoradiography. For the first time, plutonium distribution has been quantified in the human lung. The distribution of long-term retained plutonium is nonuniform, and a significant portion of plutonium was retained in pulmonary scars. In addition, a large fraction of plutonium was present in the parenchyma, where it was retained much longer than was estimated previously. The sequestration of plutonium particles in scars would greatly reduce the radiation exposure of the critical target cells and tissues for lung cancer. Thus the prolonged retention of plutonium in lung scars may not increase the dose or risk for lung cancer.     

Aminotransferase and lactate dehydrogenase activity in the blood of dogs exposed to the joint action of external gamma- and internal alpha-radiation

After the effect of external gamma-radiation (6.5 to 51.6 mC/kg) and inhaled 239Pu submicron oxide, containing 25% of 241Am, (approximately 7 to 10 kBq/kg) delivered separately and in a combination, activities of alanine-aspartate aminotransferase and lactate dehydrogenase changed in an undulatory manner tending to increase at later times; the change rating was a function of type and level of radiation as well as the time lapsed from the onset of exposure. The combined effect of gamma- and alpha-radiation did not exceed the additive effect of the two factors delivered separately.     

Organ-specific gene expressions in C57BL/6 mice after exposure to low-dose radiation

The possibility that radiation-induced alterations in gene expression are tissue specific and are related to apoptosis was examined using samples from brain, heart, lung, spleen and intestine from female C57BL6 mice after exposure to 0.2 Gy radiation. Apoptosis was the highest in spleen and intestine, moderate in lung, and absent in brain and heart. However, the mRNA expression of Trp53 and Cdkn1a (p21) after irradiation was not different among the organ types, and immunohistochemistry revealed that all the organs expressed these two proteins after irradiation. When expression patterns of 23 genes in the organs were examined by RT-PCR, neogenine, Apo1, nuclease sensitive element binding protein 1, syntaxin, cyclin G1, hNOP56, paraoxonase and glutathione peroxidase were overexpressed after irradiation in all the organs sampled, suggesting them as universal exposure markers for low-dose radiation. Sialyltransferase may be a candidate for radiation detection in spleen and intestine, which are radiosensitive organs. Because Sod1 (Cu/ZnSOD) and alphaB crystalline were expressed only in spleen, and protein tyrosine kinase and platelet membrane glycoprotein lib were expressed in both spleen and lung, these genes may also be potential markers for detection of radiation exposure, especially low-dose radiation, in these tissues. These data suggested possible tissue-specific markers of low-dose radiation exposure and suggested potential novel genetic modifiers of radiation response.     

Radiation-induced translocations in mice: persistence, chromosome specificity, and influence of genetic background

The translocation frequency response in the chromosomes of peripheral blood lymphocytes is widely used for radiation biomonitoring and dose estimation. However, this assay is based upon several assumptions that have not been rigorously tested. It is typically assumed that the translocation frequency in blood lymphocytes reflects the level of genomic damage in other hemopoietic tissues and is independent of the chromosome probe and genetic background. We conducted studies to evaluate these assumptions using mice with different genetic backgrounds. Six different whole-chromosome fluorescence in situ hybridization (FISH) probes were used to detect translocations in peripheral blood lymphocytes at multiple times after whole-body irradiation. Translocation frequencies were chromosome-independent at 6 and 16 weeks after exposure but were chromosome-dependent at 1. 5 years after exposure. Similar translocation frequencies were observed in blood, bone marrow and spleen at 1.5 years, supporting previous suggestions that genetically aberrant peripheral blood lymphocytes may derive from precursor populations in hemopoietic tissues. Translocations measured 66 h after irradiation differed among some strains. We conclude that the translocation frequency response is a complex phenotype that is influenced not only by exposure dose but also by genetic background, the choice of chromosome analyzed, and time after exposure. These results raise important considerations for the use of the FISH-based translocation frequency response for radiation dosimetry and biomonitoring.     

Relative biological effectiveness of 241Am relative to 192Ir for continuous low-dose-rate irradiation of BA1112 rat sarcomas

Sealed sources of 241Am have been developed for intracavitary irradiation of gynecological cancers. Relative to conventional isotopes (that is, 226Ra, 137Cs, 192Ir), 241Am allows for better shielding of dose-limiting normal tissues in the patient. In addition, the long half-life of 241Am (432 years) makes it an attractive isotope both for clinical use and for long-term radiobiology studies. Using a previously developed in vivo applicator system, BA1112 sarcomas on WAG/Rij Y rats were irradiated using 241Am or 192Ir at three different dose rates. Following in vivo treatment of the sarcomas with graded doses of radiation, cell survival curves were determined using an in vitro colony formation assay. The slopes of the resulting cell survival curves were observed to increase significantly as the dose rate increased from 0.30 to 0.60 Gy/h, then to decrease slightly as the dose rate increased from 0.60 to 0.95 Gy/h. The relative biological effectiveness (RBE) of 241Am relative to 192Ir was observed to increase linearly with increasing dose rate; the RBEs were 0.96 +/- 0.009, 1.09 +/- 0.12, and 1.17 +/- 0.11 at dose rates of 0.30, 0.60, and 0.95 Gy/h, respectively.     

Microdistribution and localized dosimetry of 241Am in bones of beagle dogs

The microdistribution of 241Am in selected bones of seven beagle dogs was analysed using a computer controlled microscope photometer. Four of the animals receiving between 102.5 and 165 kBq/kg were killed between 7 and 20 days after injection, and three animals receiving 32.9-34.0 kBq/kg were killed between 1300 and 1569 days. Using the photometric scanning technique, the concentrations of 241Am in several anatomical regions, as well as the specific surface activities and their variations, dose rates, accumulated radiation doses, burial depths and morphometric parameters, were derived. Dose rates to the 0-10 micron marrow band adjacent to surfaces were found to be between 8.6 and 15.7 times higher than the average skeletal dose. Accumulated radiation doses from initial deposits to lining cells were estimated to be between 87 and 252 Gy. The average burial depth in the animals killed at later times was around 8 micron. Morphometric parameters showed that radiation damage occurred in these animals, resulting in abnormal trabecular architecture. A positive correlation between specific surface activity and local turnover rates was established.     

Single inhalation exposure to 90SrCl2 in the beagle dog: hematological effects

The toxicity of 90Sr administered by the inhalation route was studied in young adult Beagle dogs exposed once to aerosols containing 90SrCl2. Due to its relatively soluble chemical form, 90Sr was rapidly translocated from lung to bone where a substantial portion was retained for a long period of time. This resulted in only a brief radiation exposure of the respiratory tract and a protracted exposure of the skeleton. The long-term retained burdens ranged from 0.037 to 4.4 MBq 90Sr/kg body wt. Dogs were subsequently observed throughout their life span. Six dogs with long-term retained burdens of 1.7 to 4.1 MBq 90Sr/kg died at less than 32 days after exposure from radiation-induced bone marrow hypoplasia. Review of hematological parameters of all dogs showed a similar, consistent, and dose-related pancytopenia in those animals having a long-term retained burden of greater than 0.37 MBq 90Sr/kg. Thrombocytopenia and neutropenia persisted in all exposed dogs through 1000 days after exposure. For reference purposes, a burden of 0.37 MBq 90Sr/kg is calculated to deliver an average radiation dose to the skeleton over 30, 100, and 1000 days after intake of 1.0, 2.8, and 17 Gy, respectively. The hematologic changes were similar to those seen in people exposed to high doses of whole-body external radiation.     

Microscopic dose distribution around PuO2 particles in lungs of hamsters,rats, and dogs

Summary Syrian hamsters, Fischer rats and Beagle dogs inhaled mono-disperse aerosols of PuO₂ and were scarificed during the first 16 days after exposure. The microscopic distribution of radiation dose and tissue-at-risk to alpha irradiation around individual particles in lung was studied using autoradiographs of lung tissue sections. The dose distributions in dogs and rats were more diffuse than in hamsters. A slightly greater tumor incidence was calculated for rats and dogs than for hamsters on the basis of dose distribution using the same dose-effect model for all three species. The small differences in tumor incidence predicted on this basis do not explain the extremely large differences in tumor incidences observed in these species after inhalation of PuO₂.Research performed under U.S. Department of Energy Contract Number DE-AC04-76EV01013. Research conducted in facilities fully accredited by the American Association for the Accreditation of Laboratory Animal Care     

Chelation therapy of incorporated plutonium-238 and americium-241: comparison of LICAM(C), DTPA and DFOA in rats, hamsters and mice

The carboxylated catechoylamide 3,4,3-LICAM(C) was tested for removal of 238Pu and 241Am from small laboratory rodents. The effectiveness of treatment was compared with that of two ligand preparations approved for clinical use: calcium-trisodium diethylenetriaminepentaacetate (DTPA) and desferrioxamine (DFOA). With early treatment and at the dosage used clinically for the decorporation of actinides with DTPA (30 mumol/kg body weight) LICAM(C) was superior to DFOA but when compared with DTPA, the effect of LICAM(C) on 238Pu was greater only in bone; as little as 1 mumol LICAM(C)/kg was as effective as 30 mumol DTPA/kg. However, in all animals treated with LICAM(C) there was a large increase in the 238Pu content of the kidney. With 241Am the effect of DTPA was always superior to that of LICAM(C). The best overall results early (1 day) after injection of 238Pu and 241Am were achieved by a combination of a single injection of LICAM(C) and DTPA with subsequent continuous administration of DTPA in drinking water. LICAM(C) affected the retention of 238Pu even if given orally; the data suggested that about 3 per cent of ingested LICAM(C) was absorbed. When the beginning of treatment was delayed, LICAM(C) became equally effective or less effective than DTPA even as far as 238Pu retention in bone was concerned, but it still increased the accumulation of 238Pu in the kidneys.