Comparative studies on the lysosomal association of monomeric 239Pu and 241Am in rat and Chinese hamster liver: analysis with sucrose, metrizamide, and Percoll density gradients of subcellular binding as dependent on time

The binding of 239Pu and 241Am in the livers of Chinese hamsters and rats was analyzed by centrifugation of a mitochondrial-lysosomal fraction in sucrose, metrizamide, and Percoll density gradients at intervals between 4 and 70 days after nuclide injection. The behavior of 239Pu and 241Am during the centrifugation experiments was very similar. In contrast to the results for rats, the median densities of the nuclide profiles from hamsters decrease with time in hyperosmolar sucrose gradients, as does the nuclide fraction liberated by addition of Triton X-100, and the nuclide profiles do not respond typically to Triton WR 1339 treatment of the animals. The results with nearly iso- osmolar metrizamide gradients, together with those for Percoll, agree with the assumption that there is an initial lysosomal association of the transuranium elements. It was concluded from the results that the major fraction of 239Pu and 241Am remained bound to typical lysosomes in rat liver, whereas those in hamster liver may be transformed into telolysosomes . Possibly, a vesicular biliary transport system for certain heavy metals, for which evidence exists in rat liver, does not occur in Chinese hamster liver.     

Association of plutonium with lysosomal, lipofuscin-like granules in Chinese hamster hepatocytes: evidence from electron microscopic and biochemical studies with 241Pu and 239Pu

This electron-microscopic-autoradiographic study was undertaken to identify the cell organelles, which bind plutonium in Chinese hamster hepatocytes at different times after injection. Female Chinese hamsters were injected intraperitoneally with 241Pu and sacrificed at time intervals of between 4 days and 35 weeks. The Chinese hamster was chosen as the experimental animal as it is a species in which there is virtually no elimination of plutonium from the liver. From the 4th day onwards beta-tracks were found over globular electron-dense structures, which were randomly distributed in the cytoplasm of the hepatocytes and strongly resembled lipofuscin bodies. Comparison of the results with those from biochemical experiments showed good agreement between the morphological and biochemical observations. At early times after injection 241Pu was also found in the hepatocyte nuclei. All the evidence suggests that in this species plutonium in hepatocytes becomes bound to lipofuscin-accumulating lysosomes, which cannot be excreted.     

Comparative assessment of the danger of inhaling 239Pu and 241Am based on the degree of leukopenia in dogs

From the analysis of the results of examination of 96 dogs it is concluded that the estimate of a relative danger of breathing transuranium radionuclides with different metabolism rates (polymeric 239Pu and monomeric 241Am), according to degree of leukopenia is more reliable when it is compared with a mean weighed dose per whole organism than the dose per individual "critical" organ.     

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.     

Influence of clay mineral type and organic matter content on the uptake of239Pu and241Am by plants

Summary The uptake of²³⁹Pu and²⁴¹Am from different clay mineral-organic matter-sand mixtures simulating contrasting soil types was examined in growth chamber experiments. The mixtures represented various combinations of organic matter (0, 5 and 10%), kaolinite (11 type) and montomorillonite (21 type) clay minerals, each at the levels of 5, 10 and 25%, and purified quartz sand (as filler).Results indicated a marked reduction in uptake of both²³⁹Pu and²⁴¹Am with increase in organic matter as well as clay content of the mixtures. The Pu Concentration Ratios (CRs) ranged from (2.5–7.0)×10^–3 in the case of kaolinite-organic matter mixtures, and from (0.9–5.5)×10^–3 in the case of montmorillonite-organic matter mixtures. The corresponding values of Am Concentration Ratios (CRs) obtained were (1.9–725.4)×10^–3 in the case of kaolinite-organic matter mixtures, and between (0.7–3.5)×10^–3 for the montmorillonite-organic matter mixtures.Reduction in the uptake of²⁴¹Am with increasing clay content was more pronounced in the montmorillonite clay-organic matter mixtures as compared to that in the case of kaolinite-organic matter mixtures. While similar qualitative reduction in²³⁹Pu CRs with increasing clay content was observed, the reduction was less marked than in the case of²⁴¹Am. The values for Am CRs were higher than the corresponding Pu CRs in kaolinite based mixtures whereas in the case of montmorillonite-organic matter mixtures Pu CRs exceeded the Am CRs.Increasing organic matter content and its interaction with both kaolinite and montmorillonite clay minerals were found to be equally effective in reducing the uptake of²³⁹Pu as well as²⁴¹Am by plants.     

Radiosensitivity of vertebral bone marrow CFU-S surviving in mice internally contaminated with239Pu or241Am

Summary The radiosensitivity of pluripotent hemopoietic stem cells was studied in ICR Swiss mice (28 g/mouse) given i.v. 198.6 kBq²³⁹Pu/kg as citrate complex or 208.6 kBq²⁴¹Am/kg as nitrate at the age of 10 weeks. The bone marrow cells were examined at the early and late phases of radionuclide contamination. To obtain data for survival curves andD ₀ of stem cells the CFU-S assay was used and the donor vertebral marrow cells were exposed to the complementary X-irradiation either early after injection to the heavily irradiated recipients or to the in vitro irradiation given before the transplantation. To determine the iron uptake in splenic erythroid progeny the recipients given marrow cells unexposed to the X-rays received 37 kBq⁵⁹Fe 6 h before they were killed and the relative activity per colony was calculated. The radiation effect of the used actinides on the bone marrow cells resulted in decreased cellularity and seriously altered both relative and absolute CFU-S numbers. The radiosensitivity of CFU-S increased in all intervals examined (D ₀ from 0.60 to 0.86 Gy, in controls 0.97 to 1.06 Gy) and was more expressed when the CFU-S were exposed to the X-rays immediately after the bone marrow cell transplantation to the heavily irradiated hosts. The stem cell pool appeared, especially at older age, to be affected also in its ability to produce erythrocytic progeny.     

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.     

Radiosensitivity and proliferative activity of vertebral CFU-S surviving in mice injected with oncogenic activities of 239Pu or 241Am

Survival, radiosensitivity and capability to produce differentiated progeny were followed in CFU-S from lumbar vertebrae of mice injected with 198.6 kBq 239Pu/kg or 208.6 kBq 241Am/kg. The CFU-S assay and 59Fe uptake into spleen colonies were used. The number of CFU-S from treated mice was significantly lower than in controls. Higher radiosensitivity of CFU-S from 239Pu- or 241Am-treated mice was demonstrated using additional exposure to 0.5 Gy X-rays 1, 24, 48, 72 hrs after cell transplantation and expressed more precisely by survival curves obtained 1 hr after the marrow cell injection. The effect of 239Pu on CFU-S was characterized by Do 0.58 Gy (n = 0.91) and that of 241Am by Do 0.64 Gy (n = 0.91); corresponding control values were Do 0.89 Gy, n = 1.11. Lower iron utilization due not only to the decreased CFU-S numbers, but also to the defective production of erythroid cells per one CFU-S was found. Complexity of radiation effect on hemopoietic stem cells was demonstrated by the present study.     

Quantitative evaluation of 90Sr, 137Cs, 239Pu, 241Am transfer from polluted soil to gastric and intestine juice of cows

Quantitative parameters of 90Sr, 137Cs, 239Pu and 241Am transfer from solid phase of soil to gastric, intestine juice of cows as well as to imitating solutions have been estimated on the base of results of laboratory incubation experiments. A prevailing role of enzyme complex and microflora of gastrointestinal tract in radionuclides transfer from solid phase of soil to solution has been shown.     

90Sr, 137Cs, 238Pu, 239+240Pu,and 241Am radionuclides in macrophytes within the Krasnensky flood plain: species specificity of concentration and distribution in phytocenosis components

The analysis of the content of radionuclides 90Sr, 137Cs, 238Pu, 239 + 240Pu and 241Am in water vegetation of flood plain reservoirs has allowed studing features of radionuclide accumulation by various species of macrophytes and revealing bioindicators of radionuclide contamination. Thus species-specificity of radionuclide accumulation can essentially change the contribution of different species to a percentage ratio of the radionuclide content in phytomass of reservoirs in comparison with fund of higher aquatic plants.     

Tissue, cellular, and subcellular distribution of 241Pu in the rat testis

The distribution and localization of 241Pu in rat testes were determined by quantitative autoradiography. Rats were given an intravenous injection of 241Pu citrate and tissues were collected 1 week later. The tissue distribution of 241Pu was determined by light microscope autoradiography. Significant concentrations of 241Pu were observed in the interstitial tissue but not in seminiferous tubules. The cellular distribution and subcellular localization of 241Pu were determined by electron microscope autoradiography. Within the interstitial tissue, 241Pu was concentrated in macrophages. There was no preferential localization of 241Pu in any other interstitial tissue components. Within interstitial tissue macrophages, 241Pu was concentrated in the lysosomal system of the cell. Other organellar compartments of the cell did not preferentially incorporate 241Pu. The association of 241Pu with the macrophage lysosomal system may explain the long retention times of Pu in testes as observed in experimental studies.     

Inhibition of spermiation in the syrian hamster using dibutyryl cyclic-AMP

Summary Young adult male Syrian hamsters were given intraperitoneal injections of 50 mg dibutyryl cyclic AMP twice daily for a period of three days. On the fourth day the animals were sacrificed and their testes were processed for light and electron microscopy. The results indicate that the mature spermatozoa were retained within the seminiferous epithelium after the stage in the seminiferous cycle in which spermiation normally occurs. The unreleased spermatozoa were ultrastructurally normal. Typical Sertoli-spermatid junctional specializations remained associated with the retained spermatozoa. These findings indicate that normally spermiation is initiated by the disappearance of the junctional specializations. In addition, the present results demonstrate that spermiation can be controlled.Supported in part by NIH Grant RR05654 and by NIH Grant P 30 HD 10202 (Morphology Core)The skillful technical assistance of Pam Duke is gratefully acknowledged     

Association of Chernobyl-derived 239+240Pu, 241Am, 90Sr and 137Cs with different molecular size fractions of organic matter in the soil solution of two grassland soils

Radiocesium is normally bound only rather weakly and unspecifically by humic substances, in contrast to the actinides Pu and Am. Recently, however, it was observed that fallout ¹³⁷Cs in the soil solution from an Of-horizon of a podzol forest soil (slightly decomposed plant material) was associated essentially only with one single size fraction of the humic substances. In deeper soil layers with well humified material (AOh-horizon), radiocesium was associated with all size fractions of the dissolved organic matter (DOM). To examine whether this unexpected behaviour is also observable for DOM isolated from other soils, we determined the association of fallout ¹³⁷Cs,⁹⁰Sr,²³⁸Pu, ^239+240Pu and ²⁴¹Am with various size fractions of DOM from in situ soil solutions isolated from two layers (0–2 cm and 2–5 cm) of two grassland soils (a soddy podzolic soil and a peat soil) within the 10 km zone of the nuclear reactor at Chernobyl (Ukraine). The four size fractions of DOM as obtained by gel filtration of the soil solution were (mean nominal molecular weight in daltons): fraction I: ≥2000, fraction II: 1300; fraction III: 560, fraction IV: inorganic compounds. The results for the well humified DOM (humus accumulation horizon of podzol, deeper layer of peat soil) showed that Pu and Am are essentially associated with the high molecular weight fractions, while Sr is present only in the `inorganic' fraction. Radiocesium is found in all the size fractions separated. A quite similar pattern was also found for Pu, Am, and Sr in the soil solution from only slightly decomposed plant material (0–2 cm of peat soil), but not for radiocesium. This radionuclide was again essentially only observable in one single low molecular weight fraction of DOM. The above results thus support our recent observations in the different horizons of a forest podzol mentioned above, even though no reason for the different binding of radiocesium by well humified soil organic matter and by only slightly decomposed plant material can be given at present. The data demonstrate, however, that information on only the total amount of a radionuclide in the soil solution will not be sufficient to interpret or predict its fate adequately in the soil. Received: 13 February 1998 / Accepted in revised form: 14 July 1998