Determination of the plutonium-239 content of the body

In order to increase the informativeness of the indirect dosimetric estimates of plutonium-239 body levels complex makers are widely used to enhance natural excretion of the radionuclide in urine, the ratio between 239Pu levels in urine and skeleton being measured. However, as the onset of chelate application is postponed its efficacy, with respect to the skeleton, sharply decreases making it impossible to obtain reliable information concerning plutonium 239 levels in bone tissues at later times.     

Comparisons of the skeletal locations of putative plutonium-induced osteosarcomas in humans with those in beagle dogs and with naturally occurring tumors in both species

Osteosarcomas occur from exposures to bone-seeking, alpha-particle-emitting isotopes, particularly plutonium. The skeletal distribution of putative 239Pu-induced osteosarcomas reported in Mayak Metallurgical and Radiochemical Plutonium Plant workers is compared with those observed in canine studies, and these are compared with distributions of naturally occurring osteosarcomas in both species. In the Mayak workers, 29% and 71% of the osteosarcomas were in the peripheral and central skeleton, respectively, with the spine having the most tumors (36%). An almost identical distribution of plutonium-induced osteosarcomas was reported for dogs injected with 239Pu as young adults. This distribution of osteosarcomas is quite different from the distributions of naturally occurring osteosarcomas for both species. In the Cooperative Osteosarcoma Study Group in humans (1,736 osteosarcomas from all ages), over 91% of the tumors occurred in the peripheral skeleton. In the Mayo Clinic group of older individuals (>40 years old), over 60% of the osteosarcomas appeared in the peripheral skeleton. The distribution of naturally occurring osteosarcomas in the canine is similar to that in the adult human. The similarities of the distributions of plutonium-associated osteosarcomas in the Mayak workers with those found in experimental studies suggest that many of the reported osteosarcomas may have been associated with plutonium exposures. These results also support the experimental paradigm that plutonium osteosarcomas have a preference for well vascularized cancellous bone sites. These sites have a greater initial deposition of plutonium, but also greater turnover due to elevated bone remodeling rates.     

Distribution of 239Pu in the skeleton of the tree shrew (Tupaia belangeri) between 15 and 50 months after injection

The macroscopic and microscopic distribution of intramuscularly injected, essentially monomeric, 239Pu was studied in the skeleton of the adult tree shrew (Tupaia belangeri). Data for the period between 15 and 50 months after injection are presented and compared with the data from earlier time points. Between 83 and 500 days after injection the nuclide content and the wet weight of the skeleton decreased to a constant level at about 55 per cent of the maximum values. The microscopic distribution has been analysed in distal femora, proximal humerus, proximal tibia and lumbar vertebra over the whole observation time; additionally at some selected time points proximal femur, femur shaft, distal humerus and distal tibia were analysed. The initial endosteal surface activity ranged from 3.8 to 5.3 Bq/cm2 and decreased to a minimum at about 1000 days after injection and increased thereafter. A similar behaviour was found for the dose rate near bone surfaces which was initially about 0.075 Gy/day on endosteal surfaces. In the deep bone and the deep marrow the dose rate was negligible, about 0.008 Gy/day and 0.001 Gy/day, respectively. The average cumulative dose 1500 days after injection was about 67 Gy on the endosteum, six times greater than the cumulative dose calculated from the mean concentration of plutonium in the whole skeleton. All values are normalized to an injected activity of 37 kBq/kg body weight. The tupaia data are discussed in relation to the available data from monkeys, dogs and rats.     

Effect of239Pu on mouse hemopoietic stem cells in different types of bone marrow cavities

Summary Repopulative activity of hemopoietic stem cells of mice given i.v. 5 kBq²³⁹Pu/mouse (166.5 kBq/kg) was followed. The activity retained was measured in the whole mouse, the skeleton and the liver. Simultaneously average cumulative skeletal dose was calculated. Quantitative parameters of the stem cell compartment and the marrow cellularity were studied in variously arranged bones (femur, pelvis, lumbar vertebra) using the exocolonizing test and cytological techniques. The effects of radiation were most marked in lumbar vertebra, less serious changes were found in pelvis and only a moderate response was present in femur. The bone marrow hemopoiesis is damaged in various bone sites to different degrees and the percentage of cells at risk appears higher in trabecular than in cortical bone.     

The ultrastructure of mouse testicular interstitial tissue containing plutonium-239 and its significance in explaining the observed distribution of plutonium in the testis.

The technique of autoradiography with Araldite-embedded sections was used to study the distribution of 239Pu in mouse testis at various times post-injection. Adjacent sections were examined with both the light microscope and electron microscope. The autoradiographs showed that from 1 week to 3 months postinjection, most 239Pu in located in interstitial tissue. The major change in distribution observed was that the early diffuse deposit in interstitial tissue is concentrated in macrophages with increasing time post-injection. This is a real change of distribution as the amount of 239Pu in mouse testis remains constant from 1 week to 3 months post-injection. Study of the ultrastructure of interstitial tissue indicated that the accumulation of 239Pu in macrophages may be brought about in two ways. First, there may be phagocytosis of dead cells containing 239Pu. Second, 239Pu may follow the transfer of waste products of hormone synthesis from Leydig cells into macrophages. The significance of these observations is discussed with regard to the deposition of 239Pu in human testis.     

Removal of monomeric 239Pu from bones and liver by liposome-encapsulated pentacin

Dependence of monomeric 239Pu removal from the liver and skeleton by liposome-encapsulated pentacine on dose and concentration of encapsulated chelate was studied in rats. It has been shown that the liposome-encapsulated pentacine (LP) removed 1.5-2.5 times as much 239Pu as free chelate (FP). Dose-effect dependences were logarithmic. The distinction between LP and FP in 239Pu removal from the liver was maximum when chelate had been used in a dose of 50 mumol/kg, with the dose effect upon injection in a large number of liposomes (200 mumol of lipids/kg) being 1.8 times as high as upon injection in smaller number of liposomes (50 mumol/kg). LP doses varying from 100 to 400 mumol/kg, there were no differences between two types of LP; with a LP dose of 400 mumol/kg its action is a bit stronger than that of the chelate. The distinction between LP and FP in 239Pu removal from the skeleton is the greatest with chelate doses exceeding 100 mumol/kg. The use of liposomes in combination with concentrated chelate solution is more effective. Possible interpretation of the features revealed are discussed.     

The subcellular distribution of 238Pu and 239Pu in primary cultures of rat hepatocytes

The subcellular distribution of 238Pu and 239Pu after incubation of primary cultures of rat hepatocytes with the citrate complex of these metals was studied, and the results were compared with data from in vivo experiments. As in vivo, the lysosomes are the principal organelles in which 238Pu and 239Pu are accumulated. In contrast to in vivo studies, 239Pu is also detectable on the pericellular membranes and in the cell nuclei, where it is predominantly bound to a high-molecular-weight component. The percentage of the total cellular 239Pu which can be recovered in the cell nuclei increased with incubation time from 10% at 1 h to nearly 30% at 5 h. Plutonium-238, an isotope with 270-fold higher specific activity than 239Pu, showed no association with the nuclei. The membrane-bound fraction of 239Pu, as determined using the exogenous chelator diethylenetriaminepentaacetic acid decreased from 30% at shorter incubation times to 15% at longer incubation periods. After incubation with 238Pu the membrane fraction and the cytosolic fraction contained higher concentrations of the radionuclide than after incubation with 239Pu.     

Self-renewal capacity of murine hemopoietic stem cells under internal contamination with239Pu and241Am

Summary The self-renewal capacity of murine pluripotent hemopoietic stem cells (CFU-S) of vertebral bone marrow was studied under conditions of short-term and long-term internal contamination with²³⁹Pu or²⁴¹Am in female mice. Measurement of the CFU-S self-renewal capacity was carried out using double transplantation assay. To evaluate the production of differentiated progeny of stem cells average erythroblast numbers/visible spleen colony and⁵⁹Fe-uptake/colony were computed. The marrow cellularity/vertebra and the number of CFU-S/vertebra were decreased and affected more by²³⁹Pu than by²⁴¹Am. The production of erythroblasts per a single CFU-S and the⁵⁹Fe-uptake/colony were reduced, similarly the numbers of secondary spleen colonies and of secondary CFU-S in primary colonies. The above changes resulting from impaired functions of surviving CFU-S were more serious with²⁴¹Am than with²³⁹Pu. The biological effects of plutonium and americium appeared independent of the phase of contamination.     

The relationship between 59Fe uptake in marrow and 239Pu deposition in bone.

The marrow in the left femur of each of 17 mice was destroyed by X-irradiation and 59Fe and 239Pu uptake into both femurs was measured 1, 3 and 7 days later. Uptake of 59Fe into marrow was depressed in the left femur 1 and 3 days after irradiation but was enhanced in the right unirradiated femur 3 days after the left femur was irradiated. There was no corresponding depression of 239Pu uptake into the left irradiated femur or enhancement into the right unirradiated femur. These results do not support the view that a functioning erythropoietic marrow is necessary for 239Pu to be deposited in bone.     

Distribution and biological effects of inhaled 239Pu(NO3)4 in cynomolgus monkeys.

Twenty male cynomolgus monkeys were exposed by inhalation either to an aerosol of 239Pu(NO3)4 to produce projected initial lung burdens of either 40, 10, or 4 kBq or to a carrier aerosol as a control. Animals died or were sacrificed at 0.01, 1, 3, 6, 12, 24, 40, and 99 months after inhalation, and the distribution and biological effects of the 239Pu were determined. The 239Pu cleared efficiently from the lungs so that less than 0.05 kBq remained at 99 months after exposure to 40 kBq. Total skeletal 239Pu activity was nearly constant after the first year, but the fraction of the body burden in skeleton at sacrifice increased with time up to 99 months because of clearance from other organs. Plutonium in the liver increased to a peak at 1 year and then decreased to about 10% of the peak value at 99 months. Plutonium in the testes was localized in the interstitial tissue with only 0.01 to 0.002% of the projected lung burden remaining in testes at 99 months after inhalation. Three animals exposed to 40 kBq of 239Pu died of radiation-related pulmonary pneumonitis and fibrosis. A primary papillary adenocarcinoma of the lung was identified in one animal exposed to 40 kBq initial lung burden and sacrificed 99 months after inhalation. The frequency of chromosome aberrations in blood lymphocytes was significantly elevated only in monkeys with projected deposits of 40 kBq of 239Pu. There was no change in aberration frequency in other exposure groups as a function of inhaled activity, time after exposure, or calculated total dose to the lungs. Only in monkeys that had marked radiation-induced pathological changes in the lung did the frequency of chromosome-type aberrations increase significantly, to a value about twice the control level. In cynomolgus monkeys, chromosome aberration frequency in blood lymphocytes is not a good indicator of radiation dose or damage from inhaled soluble plutonium.     

The induction of liver tumors by 239Pu citrate or 239PuO2 particles in the Chinese hamster

The influence of radiation dose distribution on the frequency of 239Pu-induced liver tumors was evaluated in the Chinese hamster. Different concentrations of 239Pu citrate 239PuO2 particles of known sizes were injected intravenously via the jugular vein. About 60% of the injected 239Pu citrate was deposited in the liver and 40% in the bone. The 239Pu citrate was rather uniformly distributed throughout the liver parenchyma. Injected plutonium oxide particles were taken up by the reticuloendothelial system with 90% of the body burden deposited in the liver. The 239PuO2 particles were localized in the Kupffer cells and produced nonuniform dose distributions that were dependent on particle size. There was an activity- and dose-dependent increase in the incidence of total liver parenchymal cell tumors following injection with either plutonium particles or citrate. For animals that received 14.0-, 2.7-, 0.3-, and 0.04-Gy dose to liver from 239Pu citrate the cumulative tumor incidence was 39, 32, 5, and 0%, respectively. Animals that were injected with the 0.24 micron 239PuO2 particles had doses of 42.0, 7.2, and 0.8 Gy to the liver and tumor incidences of 34, 26, and 5%, respectively. Plutonium citrate also produced hemangiosarcomas of the liver and tumors in bone and bone marrow. The latent period for liver tumor appearance in animals exposed to 239Pu citrate or 239PuO2 particles increased as the injected activity decreased. For animals injected with a similar total activity (7.4 Bq/g), the lifetime cumulative liver tumor incidence was similar for animals exposed to either 239Pu citrate (32%) or 239PuO2 (26%). There was little effect of particle size on liver tumor incidence. These data indicate that, in Chinese hamster liver, local radiation dose distribution is less important in altering tumor incidence than injected activity or average dose. However, the more uniform irradiation from 239Pu citrate administration was more effective in cancer production than the nonuniform irradiation from 239PuO2 particles.     

Quantitative microscopic studies of the distribution and retention of 239Pu in the ilium of the female CBA mouse.

Three month old female CBA mice were injected with 50 nCi kg-1 body mass of minimally polymeric 239Pu-citrate and killed at 24 hours, 10 days, 1 month and 3 months after injection. The distribution of 239Pu in the ilia of these mice was analysed using neutron-induced autoradiography of bone sections together with computer-based methods of data reduction. Results of these investigations demonstrate that while 239Pu is initially localized on bone surfaces, by 3 months after injection it is fairly uniformly distributed throughout mineral bone and its included marrow.     

Behavior and biological action of 239Pu administered intramuscularly during complexon therapy

With intramuscular injection of 239Pu nitrate the radionuclide content of the rat skeleton was higher, and at the site of injection, lower, than with injection of a polymer plutonium. The animals developed osteosarcomas (nitrate greater than polymer), and at the site of trauma, fibrosarcomas (nitrate less than polymer). After a complexon therapy the dose accretion in the skeleton and liver diminished, the life span increased, and the incidence of sarcomas decreased.     

Identification of transferrin as the principal plutonium-binding protein in the blood serum and liver cytosol of rats: immunological and chromatographic studies

An analysis of 239Pu-labelled protein complexes in serum and liver cytosol fractions prepared from rats injected intravenously with 239Pu-citrate indicated that among the possible metal-binding proteins, ferritin, transferrin and metallothionein, 239Pu was bound almost exclusively to transferrin. The method employed for the quantitative determination of 239Pu-transferrin was chromatography on immobilized rabbit anti-rat apo-transferrin. There was no evidence that transferrins from serum and liver cytosol are serologically different. The residual blood content of perfused and non-perfused liver was measured using 51Cr-labelled red blood cells and the amounts of blood-derived 239Pu and 239Pu-transferrin in the liver cytosol were calculated. The results are discussed with respect to the possible role of transferrin in the uptake of Pu into cells.     

The distribution of the radionuclides in the main components of lake ecosystems within the Chernobyl NPP exclusion zone

The results of the studies devoted to the distribution of radionuclides 90Sr, 137Cs, 238Pu, 239 + 240Pu and 241Am in 1998-2003 in main components of Glubokoe Lake and Dalekoe-1 Lake located within Krasnensky flood lands of the Pripyat River (inner exclusion zone of the Chernobyl NPP) were analysed. The data about the radionuclide content in bottom sediments, in water, in seston, in macrozoobenthos (including bivalvia molluscs), in gasteropods molluscs, in higher aquatic plants and in fish are presented.     

Efficacy of polymeric 239Pu removal by liposome-encapsulated pentacin

Liposome-incapsulated pentacine (DTPA) removes intracellular polimeric 239Pu and colloidal hydroxide of polymeric 239Pu from a rat body in the amounts 2- and 4 times, respectively, exceeding those eliminated by free DTPA. However the amount of 239Pu removed decreases sharply with increasing 239Pu hydrolysation and polymerization. It is concluded that the effectiveness of 239Pu removal depends on the physico-chemical status of the radionuclide and its interaction with the biosubstrate rather than on the amount of DTPA entering the cells.     

Intensification of the excretory flow of plutonium-239 from the body in the late stage of its metabolism

The paper deals with the effect of iron preparations on the excretion of plutonium 239 from a body at a later stage of the radionuclide metabolism. The experimental results show that oral administration of the iron preparation at a later stage of 239Pu metabolism enhances the radionuclide excretion both in urine and in faeces. On the basis of the results obtained the coefficients are calculated for 239Pu excretion in urine and faeces and for its content in the organs of deposition. This may be used for increasing the sensitivity of indirect dosimetry of plutonium-239 within the body.     

Metabolic kinetics and the biological action of 238Pu in the body of rabbits after intratracheal administration

A study was made of the distribution and biological effect of 238Pu nitrate intratracheally administered to rabbits. The skeleton and liver were the main organs in which 238Pu was secondarily deposited to make 63.5 and 12.9%, respectively, of the total amount administered. For 60 days of observation 15.3% of the amount administered were excreted in feces and urine. With 238Pu dose of 520 kBq/kg acute radiation sickness developed while at a dose of 4 kBq/kg the life span of animals did not vary from the control.     

The distribution of intratracheally administered plutonium-239 throughout the skeletal bones]

The data are presented on distribution of the intratracheally administered plutonium-239 within the skeleton bones. Plutonium was found to be distributed non-uniformly within the skeleton: it was concentrated mainly in the bones where the trabecular fraction of the bone tissue was most manifest. The results obtained were supported by the correlation analysis.