Experimental assessment of 241Am accumulation in Carassius gibelio from amphipods.

Accumulation of dietary 241Am in freshwater omnivorous fish (Carassius gibelio) was studied aiming to estimate the distribution and retention of 241Am in the fish body. Amphipods labeled with 241Am served as a food item. From 0.4 to 0.6% of ingested 241Am was retained in the fish body after short courses of feeding and depuration, the trophic transfer factor of 241Am for whole organism level was about 2∙10-4.

Plutonium Systemic Biokinetic Model for Rats.

A baseline compartmental model (relative to modeling decorporation) of the distribution and retention of plutonium (Pu) in the rat for a systemic intake is derived. The model is derived from data obtained from a study designed to evaluate the behavior of plutonium in the first 28 days after incorporation. The model is based on a recently published model of americium (Am) in rats, which incorporated a pharmacokinetic (PK)-front-end modeling approach, which was used to specify transfer to and from the extracellular fluids (ECF) in the various tissues in terms of vascular flow and volumes of ECF. In the americium model, the approach was "cell-membrane limited," meaning that rapid diffusion of americium occurred throughout all the extracellular fluids (i.e., the blood plasma and interstitial fluids), while back-end rates representing transport into and out of the cells were determined empirically. However, this approach was inconsistent with the plutonium dataset. A good fit to the data is obtained by incorporating aspects of the Durbin et al. model structure, with plutonium in plasma separated into "free" and "bound" components. Free plutonium uses a cell-membrane-limited front end as for americium. Bound plutonium uses a capillary-wall-limited front end, where transfer rates from blood plasma into the interstitial fluids are relatively slow, and must be determined either empirically or from a priori knowledge. As in the Durbin et al. model, both free and bound plutonium are available for deposition in bone. In addition, our model has some bound plutonium associated with uptake to the gastrointestinal (GI) tract. Uncertainties in transfer rates were investigated using Markov Chain Monte Carlo (MCMC). It is anticipated that this model structure of plutonium will also be useful in interpreting comparable data from decorporation studies done in experimental animals.

In vitro evidence of the influence of complexation of Pu and Am on uptake by human lung epithelial cells Calu-3.

Understanding the mechanisms involved in retention and clearance of actinides from the lungs after accidental intake is essential for the evaluation of the associated radiological risks. Although the absorption of radioelements has been shown in vivo to depend on their nature and physico-chemical properties, their mechanisms of translocation remain unknown. In this study, we have evaluated in vitro the binding and uptake by bronchial epithelial cells Calu-3 of 2 transuranic actinides, plutonium (Pu) and americium (Am), as the first steps of translocation across the pulmonary barrier. For this purpose, Calu-3 cells grown to confluence in 24-well plates were exposed to the radioelements for 24 h under various culture conditions. Two compartments were identified for the association of actinides to cells, corresponding to the membrane bound and internalized fractions. Binding of Pu was slightly higher than of Am, and depended on its initial chemical form (nitrate, citrate, colloids). Uptake of Pu and Am nitrate was higher in serum-free conditions than in supplemented medium, with an active mechanism involved in Pu internalization. Overall, our results suggest that complexation of actinides to bioligands may have an influence on their uptake by pulmonary epithelial cells, and therefore possibly on their subsequent absorption into blood.

The transfer of 241Am and 137Cs to the tissues of broilers' organs.

Data on the transfer of artificial radionuclides from the environment to the food supply is necessary for internal dose assessment. There is a necessity for expanding and improving the available information on these factors in order to make better dose models for specific scenarios. This paper describes the results of a field experiment with broiler chickens on the transfer factor (Ff) and concentration ratio (CR) for the long-term intake of 241Am and 137Cs with grass meal and soil. The broilers were divided into two groups, each group had nine subgroups and each subgroup had three broilers. The radionuclide concentrations in the feed and the thigh muscle, thigh bone, and liver of 54 broilers divided between the grass meal and soil groups were evaluated by gamma spectrometry for 241Am and 137Cs. The duration of feeding with "contaminated" sources ranged between 1-70 days. The equilibrium stage of 241Am in muscle and bone occurs on the 1st and 40th day, respectively; for 137Cs in muscle- 30th days of intake and for liver and bone- 7th days. For 241Am, the liver did not reach equilibrium stage during the 70 days of intake. Ff of 137Cs in the "forage-muscle" and "soil-muscle" systems were determined as 1.9±0.3 and 0.18±0.05; Ff of 241Am in the "soil-muscle" system was-7.5×10-5.

Americium Systemic Biokinetic Model for Rats.

In this work, a baseline compartmental model of the distribution and retention of americium in the rat for a systemic intake was derived. The model was derived from data obtained from a study designed to evaluate the behavior of americium in the first 28 days after incorporation. A pharmacokinetic (PK)-front-end modeling approach was used to specify transfer to and from the extracellular fluids (ECF) in the various tissues in terms of vascular flow and volumes of ECF. Back-end rates representing transport into and out of the cells were determined empirically. Uncertainties in transfer rates were investigated using Markov chain Monte Carlo (MCMC). The combination of PK-front-end model and the back-end model structure used allowed for extrapolation to the earliest times with small uncertainty. This approach clearly demonstrated the rapid transfer of material from ECF to liver and bone. This model provides a baseline for modeling the action of decorporation agents, such as DTPA.

Uptake, depuration, dose estimation and effects in zebrafish exposed to Am-241 via dietary route.

Zebrafish were chronically exposed to Am-241, an alpha-emitting radionuclide via daily consumption of an enriched artificial diet. Am-241 uptake was quantified in Danio rerio after 5 and 21 days of exposure via daily Am-spiked food ingestion and after 21 days of exposure followed by 5 days of depuration. Americium accumulates mostly in digestive tract, muscle, rest of the body but the accumulation levels and trophic transfer rate (0.033-0.013%) were low. Corresponding cumulative doses were calculated for the whole body (9 mGy) and for the digestive tract (42 mGy) with internal alpha radiation contributing to more than 99% of the total dose. Genotoxic effects (gamma-H2AX assay) and differential gene expressions of main biological functions were examined. Although fish were exposed to a low dose rate of 13 µGy h-1, DNA integrity and gene expression linked to oxidative stress, hormonal signaling and spermatogenesis were altered after 21 days of Am-241 exposure. These results underline the higher toxicity of alpha emitter Am-241, as compared to other studies on gamma radiation exposure.

Radioecological aftermath: Maternal transfer of anthropogenic radionuclides to shark progeny is sustained and enhanced well beyond maternal exposure.

Cartilaginous dogfish Scyliorhinus canicula continued to transfer four anthropogenic radionuclides (65Zn, 60Co, 134Cs and 241Am) to their eggs for over six months, after two months of continued maternal exposure to radio-labelled food. Unexpectedly, rates of radionuclide transfers to eggs and their yolk & embryo during maternal depuration were equivalent for 60Co and 241Am, or even enhanced for 65Zn and 134Cs by factors of c.200-350%, over two-three months, compared to their maximal transfer rates at the end of the maternal uptake phase. These rates of maternal transfer of radionuclides to yolk & embryo were positively associated with their previously determined efficiencies of assimilation (AE) from ingested radio-labelled food. Thus progeny may be more exposed via maternal transfer to those radionuclides which have greater rates of maternal assimilation from food. As maternal depuration continued beyond 60-80 up to 180-200 days the transfers of all four radionuclides to eggs did diminish but were still substantial at mean values of 18% for 241Am, 17% for 134Cs and 9 and 8% for 60Co and 65Zn, respectively. In the yolk & embryo the mean rates of transfer over this period were further reduced for 241Am (13.5%), 60Co (2.5%) and 65Zn (5.8%), but were still appreciable for 134Cs at 56%. These results for S. canicula have demonstrated a potential enhanced radiological risk of extended duration due to the particular biokinetics of maternal transfer in this species. This study draws further attention to the current paucity of knowledge about the maternal: progeny transfer pathway, particularly in the context of the known heightened radio-sensitivity of early life stages in fish and other vertebrates, compared to later life stages.

Decorporation of Pu/Am Actinides by Chelation Therapy: New Arguments in Favor of an Intracellular Component of DTPA Action.

Diethylenetriaminepentaacetic acid (DTPA) is currently still the only known chelating drug that can be used for decorporation of internalized plutonium (Pu) and americium (Am). It is generally assumed that chelation occurs only in biological fluids, thus preventing Pu/Am deposition in target tissues. We postulate that actinide chelation may also occur inside cells by a mechanism called "intracellular chelation". To test this hypothesis, rats were given DTPA either prior to (termed "prophylactic" treatment) or belatedly after (termed "delayed" treatment) Pu/Am injection. DTPA decorporation efficacy was systematically tested for both plutonium and americium. Both prophylactic and delayed DTPA elicited marked decreases in liver Pu/Am. These results can be explained by chelation within subcellular compartments where DTPA efficacy increased as a function of a favorable intracellular DTPA-to-actinide molar ratio. The efficacy of intracellular chelation of liver actinides decreased with the delay of treatment. This is probably explained by progressive actinide binding to the high-affinity ligand ferritin followed by migration to lysosomes. Intracellular chelation was reduced as the gap between prophylactic treatment and contamination increased. This may be explained by the reduction of the intracellular DTPA pool, which declined exponentially with time. Skeletal Pu/Am was also reduced by prophylactic and delayed DTPA treatments. This decorporation of bone actinides may mainly result from extracellular chelation on bone surfaces. This work provides converging evidence for the involvement of an intracellular component of DTPA action in the decorporation process. These results may help to improve the interpretation of biological data from DTPA-treated contamination cases and could be useful to model DTPA therapy regimens.

Microbial mobilization of plutonium and other actinides from contaminated soil.

We examined the dissolution of Pu, U, and Am in contaminated soil from the Nevada Test Site (NTS) due to indigenous microbial activity. Scanning transmission x-ray microscopy (STXM) analysis of the soil showed that Pu was present in its polymeric form and associated with Fe- and Mn- oxides and aluminosilicates. Uranium analysis by x-ray diffraction (µ-XRD) revealed discrete U-containing mineral phases, viz., schoepite, sharpite, and liebigite; synchrotron x-ray fluorescence (µ-XRF) mapping showed its association with Fe- and Ca-phases; and µ-x-ray absorption near edge structure (µ-XANES) confirmed U(IV) and U(VI) oxidation states. Addition of citric acid or glucose to the soil and incubated under aerobic or anaerobic conditions enhanced indigenous microbial activity and the dissolution of Pu. Detectable amount of Am and no U was observed in solution. In the citric acid-amended sample, Pu concentration increased with time and decreased to below detection levels when the citric acid was completely consumed. In contrast, with glucose amendment, Pu remained in solution. Pu speciation studies suggest that it exists in mixed oxidation states (III/IV) in a polymeric form as colloids. Although Pu(IV) is the most prevalent and generally considered to be more stable chemical form in the environment, our findings suggest that under the appropriate conditions, microbial activity could affect its solubility and long-term stability in contaminated environments.

Effect of low-dose ionizing radiation on luminous marine bacteria: radiation hormesis and toxicity.

The paper summarizes studies of effects of alpha- and beta-emitting radionuclides (americium-241, uranium-235+238, and tritium) on marine microorganisms under conditions of chronic low-dose irradiation in aqueous media. Luminous marine bacteria were chosen as an example of these microorganisms; bioluminescent intensity was used as a tested physiological parameter. Non-linear dose-effect dependence was demonstrated. Three successive stages in the bioluminescent response to americium-241 and tritium were found: 1--absence of effects (stress recognition), 2--activation (adaptive response), and 3--inhibition (suppression of physiological function, i.e. radiation toxicity). The effects were attributed to radiation hormesis phenomenon. Biological role of reactive oxygen species, secondary products of the radioactive decay, is discussed. The study suggests an approach to evaluation of non-toxic and toxic stages under conditions of chronic radioactive exposure.

Species-dependent chelation of (241)Am by DTPA Di-ethyl ester.

Diethylenetriaminepentaacetic acid (DTPA) is an FDA-approved chelating agent for enhancing the elimination of transuranic elements such as americium from the body. Early access to therapy minimizes deposition of these radionuclides in tissues such as the bone. Due to its poor oral bioavailability, DTPA is administered as an IV injection, delaying access. Therefore, a diethyl-ester analog of DTPA, named C2E2, was synthesized as a means to increase oral absorption. As a hexadentate ligand, it was hypothesized that C2E2 was capable of binding americium directly. Therefore, the protonation constants and americium stability constant for C2E2 were determined by potentiometric titration and a solvent extraction method, respectively. C2E2 was shown to bind americium with a log K of 19.6. The concentrations of C2E2, its metabolite C2E1, and DTPA required to achieve effective binding in rat, beagle, and human plasma were studied in vitro. Dose response curves for each ligand were established, and the 50% maximal effective concentrations were determined for each species. As expected, higher concentrations of C2E2 were required to achieve the same degree of binding as DTPA. The results indicated that chelation in beagle plasma is more representative of the human response than rats. Finally, the pharmacokinetics of C2E2 were investigated in beagles, and the data was fit to a two-compartment model with elimination from the central compartment, along with first-order absorption. Based on the in vitro data, a 100 mg kg dose of C2E2 can be expected to have an effective duration of action of 3.8 h in beagles.

Orally administered DTPA penta-ethyl ester for the decorporation of inhaled (241)Am.

Diethylenetriaminepentaacetic acid (DTPA) is an effective decorporation agent to facilitate the elimination of radionuclides from the body, but its permeability-limited oral bioavailability limits its utility in mass-casualty emergencies. To overcome this limitation, a prodrug strategy using the penta-ethyl ester form of DTPA is under investigation. Pharmacokinetic and biodistribution studies were conducted in rats by orally administering [(14) C]DTPA penta-ethyl ester, and this prodrug and its hydrolysis products were analyzed as a single entity. Compared with a previous reporting of intravenously administered DTPA, the oral administration of this prodrug resulted in a sustained plasma concentration profile with higher plasma exposure and lower clearance. An assessment of the urine composition revealed that the bioactivation was extensive but incomplete, with no detectable levels of the penta- or tetra-ester forms. Tissue distribution at 12 h was limited, with approximately 73% of the administered dose being associated with the gastrointestinal tract. In the efficacy study, rats were exposed to aerosols of (241) Am nitrate before receiving a single oral treatment of the prodrug. The urinary excretion of (241) Am was found to be 19% higher than with the control. Consistent with prior reports of DTPA, the prodrug was most effective when the treatment delays were minimized.

Detoxification of AM-241 solutions by humic substances: bioluminescent monitoring.

The study addresses the effect of humic substances on marine luminous bacteria Photobacterium phosphoreum exposed to Am-241 (3,000 Bq L(-1), water solution). Luminescent intensity of the bacteria was applied as a marker of their physiological activity. Humic substances have been found to reduce the effect of Am-241 on luminescence, decrease damage to cells, and change distribution of Am-241 between bacterial cells and intercellular media. It was shown that water-soluble humic substances, being products of natural transformation of organic substances in soil and bottom sediments, can serve as protecting agents for water microorganisms exposed to alpha radionuclides.

Microdistribution of 241Am in structures of submerged macrophyte Elodea canadensis growing in the Yenisei River.

A submerged macrophyte of the Yenisei River, Elodea canadensis, was used to study the microdistribution of the artificial radionuclide (241)Am among different components of the plant. The total amount of (241)Am added to the experimental system was 1850+/-31 Bq/L. The total amount of (241)Am accumulated by the plants was 182 Bq per sample, or 758,333+/-385 Bq/kg dry mass. It has been found that the major portion of (241)Am accumulated by E. canadensis, up to 85%, was bound to solid components of the cells. It is observed that the microdistribution of (241)Am within different components of the submerged plant E. canadensis was not uniform. (241)Am distribution vary depending on the age of the leaf blades, the state of the cells and morphological features of the plant stem.

Accumulation of (241)Am by suspended matter, diatoms and aquatic weeds of the Yenisei River.

In this work we experimentally estimated the capacities of the key components of the Yenisei River (Russia): particulate suspended matter (seston), diatom microalgae, and submerged macrophytes for accumulating (241)Am from water. In our experiments large particles of seston (>8mum), comparable in size with diatoms, took up most of americium from water. The accumulation of americium by isolated diatom algae (Asterionella formosa and Diatoma vulgare) was lower than by total seston. The concentration factors (CFs) of (241)Am for seston of the Yenisei River in our experiments were (2.8-6.9).10(5); for diatoms - (1.5-4.2).10(4). The CFs for aquatic plant Elodea canadensis were within the same order of magnitude as those for diatoms. Activity concentration and CFs of (241)Am were nearly the same in experiments under dark and light conditions. This is indicative of an energy independent mechanism of americium uptake from the water by diatoms and submerged macrophytes.

[Transuranium elements on the Belarus territory].

The isotope composition and the dynamics of its change in Chernobyl falling-outs in the Belarus territory are studied. The levels of the content of transuranium elements in the basic components of ecosystems such as air, ground, water, plants and animals are quatified. The features of the inflow of transuranium elements into living organisms are determined.

[Radioecological studies of freshwater mollusks in the Chernobyl accident exclusion zone].

Species-specificity and dynamics of 90Sr, 137Cs and some transuranic elements accumulation in bivalve and gastropod freshwater molluscs of the Chernobyl exclusion zone during 1997-2008 was analyzed. The results of radiation dose and chromosome aberration rate estimation and the analysis of hemolymph composition of freshwater snail (Lymnaea stagnalis L.) was produced. The absorbed dose rate was registered in the range of 0.3-85.0 microGy/h. In closed water bodies the heightened chromosome aberration rate (up to 27%) in embryo tissues, and also the change of haematological indexes for the adult individuals of snails was registered.

[The accumulation of 241Am by suspended matter of the Yenisei River].

Laboratory experiments were performed to determine parameters of accumulation of 241Am by suspended particulate matter (seston) of the Yenisei River, with particles larger than 1 microm, and the diatoms A. formosa and D. vulgare. Concentration factors for seston were (2.8-4.1) x 10(5) and for diatoms--(1.5-4.2) x 10(4). As phytoplankton's contribution to the seston mass is rather small (< 10%), we assume that suspended matter contains other particles similar in size to the Yenisei River phytoplankton, which make larger contribution to 241Am concentration of seston than the studied algae. No energy-dependent accumulation of americium by algae was detected in the experiments. Addition of dissolved organics and hydrogen carbonates led to a lower uptake of 241Am from the Yenisei water by seston.