Historical overview of the project "Sunshine" in Belgium.

The project "Sunshine" was originally focused on the study of strontium 90 from the world-wide radioactive fallout and its effects on man. It was one of the most important projects the USAEC had ever had. It was being conducted as a scientific study, the primary purpose of which was to discuss the scientific truth and present the facts publicly. The implementation of the Project in Belgium started in 1958 and developed into two main directions: a) Environmental survey and b) Experimental research in both natural and controlled conditions. The paper described the characteristics of the seven stations "Sunshine" distributed in the typical agricultural regions of the country, their sampling, the analytical procedures used and the results obtained (relationship rain/grass; milk/grass; the observed ratio etc.). Experimental researches performed in natural conditions aimed to investigate the influence of the yield of the grass/crops on the direct retention of the deposited fission products; other experiments watched the transfer of radiostrontium and radiocesium from soils to plants; several experiments on artificially contaminated pastures grazed by cows allowed to clarify the importance of such factors as the methods of feeding and types of pastures for the transfer of major radionuclides to milk. Experimental researches conducted in controlled conditions dealt with such topics as study on the retention of strontium 90 in soil, studies on foliar contamination by radiostrontium and radiocesium, absorption and distribution in plants of Sr and Ca, discrimination between Sr and Ca during the transfer from feed to cow milk, study on the contamination by the major fission products of the milk products. Finally tentative countermeasures were tested: application of significant amounts of stable strontium in contaminated soils and influence of alginate on radiostrontium absorption from contaminated milk to piglets.

On the pathogenesis of atherosclerosis: enzymatic transformation of human low density lipoprotein to an atherogenic moiety.

Combined treatment with trypsin, cholesterol esterase, and neuraminidase transforms LDL, but not HDL or VLDL, to particles with properties akin to those of lipid extracted from atherosclerotic lesions. Single or double enzyme modifications, or treatment with phospholipase C, or simple vortexing are ineffective. Triple enzyme treatment disrupts the ordered and uniform structure of LDL particles, and gives rise to the formation of inhomogeneous lipid droplets 10-200 nm in diameter with a pronounced net negative charge, but lacking significant amounts of oxidized lipid. Enzymatically modified LDL (E-LDL), but not oxidatively modified LDL (ox-LDL), is endowed with potent complement-activating capacity. As previously found for lipid isolated from atherosclerotic lesions, complement activation occurs to completion via the alternative pathway and is independent of antibody. E-LDL is rapidly taken up by human macrophages to an extent exceeding the uptake of acetylated LDL (ac-LDL) or oxidatively modified LDL. After 16 h, cholesteryl oleate ester formation induced by E-LDL (50 micrograms/ml cholesterol) was in the range of 6-10 nmol/mg protein compared with 3-6 nmol/mg induced by an equivalent amount of acetylated LDL. At this concentration, E-LDL was essentially devoid of direct cytotoxic effects. Competition experiments indicated that uptake of E-LDL was mediated in part by ox-LDL receptor(s). Thus, approximately 90% of 125I-ox-LDL degradation was inhibited by a 2-fold excess of unlabeled E-LDL. Uptake of 125I-LDL was not inhibited by E-LDL. We hypothesize that extracellular enzymatic modification may represent an important step linking subendothelial deposition of LDL to the initiation of atherosclerosis.

Technetium absorption and turnover in monogastric and polygastric animals.

Technetium (Tc) released into the environment can reach animals in various chemical forms: as pertechnetate (TcO-4) in drinking water or deposited on the surface of vegetables and forage plants, or as Tc bioincorporated into plants and associated with various plant constituents. In addition to being influenced by chemical speciation in the diet, absorption, metabolism, and retention of Tc in animals are modified by the treatment that the alimentary bolus undergoes during its passage through the gastrointestinal tract. This behavior differs markedly between polygastric and monogastric animals. We have, therefore, studied the fate of 99mTc given in the diet either as TcO-4 or bioincorporated into maize in rats (as an example of a monogastric animal) and in sheep (as an example of a polygastric animal). Urine and feces were collected and assayed for Tc activity by gamma spectrometry. Animals were sacrificed at different times after contamination, and the Tc content of tissues was determined. The pattern of absorption, excretion and, to a certain degree, of organ distribution and retention depended on animal species and species of Tc administered. Excretion was by feces and urine, and several metabolic components could be discerned. A component of very short half-time in urine suggests that newly absorbed Tc is more readily excreted than that already bound by tissues. The highest tissue concentrations were found in the thyroid. Retention of Tc was, however, most pronounced in bone and skin. Hair contains considerable amounts of Tc and may serve as a bioindicator of Tc contamination.

Assessment of dose to man from releases of 99Tc in fresh water systems.

The objective of this paper is to evaluate the dose to man from releases of 99Tc in a fresh water system and to identify the biospheric transfer parameters to which the total dose is the most sensitive. Only internal exposure is taken into account, as the external irradiation leads to a negligible dose contribution. Two release modes were considered: continuous (routine) releases and accidental releases. The concentrations in the biospheric compartments subsequent to routine releases were calculated according to International Atomic Energy Agency procedures. For the accidental releases, a more dynamic approach was adopted, especially for the milk and meat compartments. A routine-release scenario typical for the Mol site has been applied, and the biospheric compartment leading to the highest dose contribution was shown to be the irrigated grain. The biospheric transfer parameters to which the first-year doses were the most sensitive consisted mainly of the mass interception factor for grain and the milk transfer factor. The doses in following years were very dependent on the value of the root zone removal rate. The accidental-release scenario resulted in committed dose equivalent that are strongly influenced by the time of year at which the release occurs.

Toxicity of 99Tc: can it represent a risk to man?

The movement through the environment and the toxicity to plants of 99Tc have been studied. However, information is scanty on 99Tc toxicity to mammals even though this is the decisive criterion for assessing the consequences of releases of Tc. A critical tissue could be the thyroid because of its preferential accumulation of Tc and the developing organism because of its greater radiosensitivity; moreover, this toxicity might be enhanced under conditions of a low iodine (I) diet. These questions were studied in rats given large amounts of 99Tc in either a normal or an I-deficient diet for several months starting 2 wk before mating. Newborns were continued on these diets after weaning. The parameters determined were: occurrence of pregnancy and litter size, triiodothyronine and thyroxin in serum, uptake of 131I by the thyroid 24 h after injection, histology of the thyroid and concentration of Tc in tissues of the mothers and their decendants. Thyroid damage, as well as the effects on pregnancy, could be observed after amounts of 10 micrograms Tc g-1 food. Iodine deficiency only slightly influences the 99Tc toxicity. The chemical and radiological toxicity of 99Tc to rats is small. Consequently, it seems unlikely that contamination levels in the environment would ever reach levels that could lead to serious non-stochastic effects, even in the developing organism.

The transfer of tritium-labeled organic material from grass into cow's milk.

Two lactating cows were given tritiated hay containing organically bound tritium (OBT) only for about 4 weeks. Tritium activity was determined in milk fat, casein, lactose, milk water, and whole milk. In one cow, milk was sampled for approximately 450 days, covering two lactation periods. At steady state, specific tritium activities in casein, lactose, and milk water were 58, 10, and 11%, respectively, of those in milk fat. Some OBT was converted into THO during catabolism and entered the body water pool. This 3H source accounted for nearly 40% of tritium in lactose, but in casein and milk fat about 97% of tritium was derived from ingested OBT. Comparison of the specific activity of milk constituents with the specific activity of ingested hay showed the following values: 0.84 for milk fat, 0.49 for casein, 0.05 for lactose, 0.10 for milk water. About one-half of the tritium transferred to milk was found in organic milk constituents and the other half in milk water. Decrease of tritium activity with time could be represented by three components with different half-lives for the organic milk constituents. Those for milk fat and casein were quite similar, with a slow component of nearly 3 months.

Metabolism of 51Cr, 54Mn, 59Fe and 60Co in lactating dairy cows.

Different radionuclides which can be released by nuclear installations (Na2(51) CrO4 , 54MnCl2 , 59FeCl3 and 60CoCl2 ) were given to lactating dairy cows either orally or by intravenous (i.v.) injection. Excretion into feces and urine as well as secretion into milk were followed for several weeks. Distribution of activity in organs was determined at sacrifice 102 days after oral and 70 days after i.v. administration. After ingestion, excretion of chromate followed a three-term exponential function with half-lives of 0.88, 3.7 and 26 days. Intestinal absorption was on the order of 0.1-0.2%. About 63% of injected chromate was excreted into urine, about 18% into feces and about 3.6% into milk. Orally administered chromate was concentrated in liver, intestine and spleen. The transfer coefficient into milk was about 1 X 10(-5) days/1. Less than 1% of an ingested dose of manganese was absorbed. Excretion occurred mainly into feces and followed a three-term exponential function. Very little manganese was excreted into urine or secreted into milk. Manganese is concentrated in brain, pancreas, kidney and heart but the differences in concentration are small. The body burden is, therefore, mainly determined by manganese in muscle, skin and bone. The transfer coefficient of manganese into milk is about 3 X 10(-5). Excretion of iron into feces after oral administration follows a three-term exponential function with a small component having a half-life of 72 days. Intestinal absorption is on the order of 0.5-2% of the dose. After i.v. administration, 7% of the dose is excreted in the feces displaying two components of turnover. Very little iron is excreted into urine. Secretion into milk also follows a two-term exponential function. The transfer coefficient of iron into milk is about 3 X 10(-5). About 0.6% of an oral and about 94% of an i.v. dose were recovered from the cows at sacrifice. Most activity was present in blood, liver and spleen. Excretion of radiocobalt into feces after oral administration is described by a three-term exponential function, while excretion into milk and urine is described by two-term exponential functions. Long-lived components in urine and milk represent about 5-10% of the activity absorbed. Only 1-2% of an oral cobalt dose is absorbed and 0.05% of an oral and 5.85% of an i.v. dose is recovered from the cow at sacrifice where concentrations are highest in kidney and thyroid. The body burden depends mainly on cobalt in muscle, skin and bone. The transfer coefficient of cobalt into milk is about 7.5 X 10(-5).

Study of the distribution and biological effects of 3H in the algae Acetabularia, Chlamydomonas and Porphyra.

Freshwater and marine algae are particularly useful for studying the radioactive contamination of aquatic systems. Acetabularia, Chlamydomonas and Porphyra were used to investigate the uptake and eventual biological effects of tritium. When the Acetabularia are grown in the presence of tritiated water, a significant amount of 3H is incorporated in the total nucleic acids and protein fraction. Chloroplasts of Acetabularia were isolated from whole cells and their DNA purified by the agarose procedure, before radioactivity analysis: a significant amount of 3H was incorporated into the chloroplast genome. Chlamydomonas was grown on minimal medium containing increasing concentrations of tritiated water. The increase in cell number was checked by microscope counting. The generation time was 9.6 h and seemed not affected even by the highest 3H concentration. Parallel experiments have shown that an appreciable amount of 3H was incorporated into the total organic matter of the plants. In the case of Porphyra, it was found that a very low level of 3H was incorporated into the total DNA of the plant.

Distribution of tritium in the different organs of calves and pigs after ingestion of various tritiated feeds.

A research programme on the transfer of tritium in the food chain has been in progress for several years on the experimental farm of the Nuclear Energy Research Center at Mol. The studies reported here are related to the distribution of tritium in the organs of farm animals contaminated in various ways. Two young male calves ingested tritiated milk; the daily intake of 3H-organic form was about 15 muCi for each calf and the total activity ingested until the sacrifice was 482 muCi. Three male pigs from the same litter and about 7 weeks old were used for each experiment on the administration of tritium under different forms: (a) single intraperitoneal injection of 39.3 mCi HTO (P2, P3, P4). (b) daily ingestion of 28.4 muCi HTO. The total activity ingested was respectively 569 muCi (P5) and 766.8 muCi (P6). (c) ingestion of tritiated potatoes. The total activity ingested was respectively 21 muCi (P8), 40.3 muCi (P9) and 48.1 muCi (P10). (d) ingestion of tritiated milk powder. The total activity ingested was respectively 60.6 muCi (P13), 110.4 muCi (P11) and 154.5 muCi (P12). After slaughtering of each animal various organs were removed and analyzed for the 3H content in the tissue water and in the organic matter. We could verify that the chemical form of 3H present in the food is of great importance for the incorporation of 3H in the organic matter of the animal organs. The total incorporation increases by a factor 5.6 when 3H is ingested as tritiated milkpowder by pigs as compared to HTO and with a factor 15 for calves. When tritiated potatoes were ingested by pigs a factor 15.6 was found. The transfer of 3H from HTO and milk feed ingested in the organic fraction of organs is lower for pig than for calf. When we consider the 3H in the tissue water of organs the specific activity (SA) is a little lower than the SA of ingested HTO and after ingestion of tritiated milk feed the activity is very low and no difference due to the species is found. After fractionation of liver and spleen tissue following the technique of Schmidt--Thannhauser radioactivity was found in all liver and spleen constituent lipids--RNA--DNA and proteins, but after isolation and purification of DNA following the original methods, we have not been able to demonstrate that tritium is really incorporated into DNA molecules of a non-dividing organ such as the liver nor of an actively dividing organ such as the spleen.