THE TOXICOLOGY OF RADIOACTIVE SUBSTANCES VOLUME 4 Thorium-232 and Uranium-238 Edited by A. A. LETAVET and Ε. B. K U R L Y A N D S K A Y A Translated by A. CROZY Translation edited by G. W. D O L P H I N U N I T E D K I N G D O M A T O M I C E N E R G Y A U T H O R I T Y Authority Health and Safety Branch Radiological Protection Division, Harwell 1966 P E R G A M O N PRESS O X F O R D · L O N D O N . E D I N B U R G H · N E W Y O R K T O R O N T O · S Y D N E Y · P A R I S · B R A U N S C H W E I G Pergamon Press Ltd., Headington Hill Hall, Oxford 4 & 5 Fitzroy Square, London W,l Pergamon Press (Scotland) Ltd., 2 & 3 Teviot Place, Edinburgh 1 Pergamon Press Inc., Maxwell House, Fairview Park, Elmsford, New York 10523 Pergamon of Canada Ltd., 207 Queen's Quay West, Toronto 1 Pergamon Press (Aust.) Pty. Ltd., 19a Boundary Street, Rushcutters Bay, N.S.W. 2011, Australia e Pergamon Press S.A.R.L., 24 rue des Ecoles, Paris 5 Vieweg & Sohn GmbH, Burgplatz 1, Braunschweig Copyright © 1970 P E R G A M O N P R E S S L T D . First English edition 1970 Al Rights Reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, elec- tronic, mechanical, photocopying, recording or otherwise, without the prior permission of Pergamon Press Ltd. Library of Congress Catalog Card No. 61-9783 This is a translation of the original Russian published in 1964 by Medgiz, Moscow Printed in Hungary 08 013413 0 C E R T A I N ASPECTS OF THE TOXICOLOGY OF I N S O L U B L E C O M P O U N D S OF T H O R I U M - 2 3 2 A N D U R A N I U M - 2 3 8 Ε. B. KURLYANDSKAYA STUDY of the toxicity of the natural radioactive substances thorium-232 and uranium-238 is an important practical and theoretical task. Theoret- ical interest in the problems of the toxicology of thorium and uranium is determined by the dual nature of the biological effects of these elements on entry into the body. The point is that early biological effects produced by these substances are the result of their genuine chemical toxicity, whereas the late ones, as a rule, depend on the radioactivity of the compounds of thorium and uranium. The literature contains quite a considerable volume of information characterizing the toxic properties of soluble and insoluble compounds of these elements. Thus, McClinton etal. (1948) consider that the L D 50 of soluble thorium nitrate on intraperitoneal injection into rats is 68 mg/kg (calcu- lated for thorium) and its tolerated dose 48-6 mg/kg. According to Wata- nabe (1957) the mean lethal dose of thorium chloride injected intravenously into mice is 44-3 mg/kg and the tolerated dose 5 mg/kg. The insoluble salts of thorium-232 are distinguished by low toxicity as indicated by the considerable clinical material accumulated as a result of use of colloidal thorium dioxide (Thorotrast) for radiodiagnosis. At late stages after administration, the radiotoxic properties of Thorotrast became manifest, producing a rather high incidence of neoplasma in the parenchy- matous organs rich in reticulo-endothelial elements. The same also applies to uranium compounds. The high toxicity of the nitrates, chlorides and other soluble compounds of uranium is known and was already described in 1889 (Woroschilsky, 1889). According to Haven and Hodge (1949) the intraperitoneal rat L D 50 for uranium nitrate is 2-37 mg/kg (1 - 1 2 mg/kg calculated for uranium) and the equivalent for rabbits 2 mg/kg (0-95 mg/kg for uranium). These .findings indicate the high toxicity of soluble com- pounds of uranium. Its insoluble compounds (UO2, UeOg) are considerably less toxic, according to Dygert et al (1949) . Pathological lesions are also 1 2 The Toxicology of Radioactive Substances seen at late times after administration of these compounds, which may be connected with the alpha-activity of uranium. Despite the considerable importance of thorium and uranium as will be discussed in detail in the relevant papers in the present volume and de- spite the attention which in the last few years has been paid to the toxicology of these elements, we still lack sufficiently convincing data on the behaviour of their various compounds, their biological effects and also the late se- quelae of their administration. This is especially the case for the insoluble compounds. The present collection presents material from experimental investiga- tions carried out in the radiotoxicology laboratory of the Institute of La- bour Hygiene and Occupational Diseases, U.S.S.R. Academy of Medical Sciences, and devoted to certain aspects of the toxicology of insoluble com- 2 3 2 pounds of these elements, namely, thorium dioxide ( Th02) and uranous- 2 3 8 uranic oxide ( U 0 ) . 3 8 To establish the toxicity of thorium dioxide, Yan Syao-Shan compared it with the soluble compounds thorium nitrate and chloride. It was found that the most toxic is thorium nitrate, the L D 5 03/ o of which on intraperito- neal injection in mice is 370-8 mg/kg (calculated for thorium). According to Traikovich, intracardiac administration of 1 mg/kg thorium nitrate causes instant death of rats as a result of embolism by coagulated proteins. The least toxicity is shown by thorium dioxide, the minimum lethal dose of which is about 2000 mg/kg. The comparative data on the behaviour of soluble and insoluble com- pounds of thorium for different routes of administration are of definite interest. Thus, the investigations carried out by Traikovich show that on intracardiac administration of thorium citrate to rats the greatest amount of thorium is found in organs rich in reticuloendothelial elements—the liver and spleen (17 and 3 per cent of the thorium administered). For the intraperitoneal, intramuscular and oral routes, 53-75 per cent of the amount found in the body was detected in the bones (about 2 per cent of that administered). A large amount of thorium was at the site of adminis- tration apparently connected with the formation of poorly soluble com- plexes which are fixed there for a long time. Different behaviour is shown by thorium dioxide. As shown by the in- vestigations of Yan Syao-Shan, thorium dioxide is retained for a long time in the lungs on intratracheal and in the abdominal cavity on intraperito- neal administration. Thus, in the early stages after administration of thorium dioxide from 68 to 73 per cent is found in the lungs and after 21 months up to 30 per cent of that administered. Only in the first few days after intratracheal administration was thorium dioxide found in the tra- chea, lymph nodes, gastro-intestinal tract, liver, bones and kidneys. Toxicology of Insoluble Compounds of Thorium-232 and Uranium-238 3 Up to 75 per cent of the thorium dioxide administered was found in the abdominal cavity 15 months after intraperitoneal injection. Some was found in the mesenterial lymph nodes and also about 0-33-0-51 per cent of the amount administered was in the bones. These findings show that a certain part of the thorium dioxide adminis- tered is dissolved in body fluids. Possible evidence of this is the presence of thorium in the bones and also its excretion in the urine. Some of the thorium dioxide may be removed by the phagocytes and is held in the reti- culoendothelial tissue. But the fraction of thorium dioxide retained by the organs and the tissues is negligible as compared with the amount fixed at the site of administration. In line with the low true toxicity of thorium dioxide the changes in the rat body at early times (during 10-12 months) after intratracheal and intraperitoneal administration are slight and differ little from those observed in control animals. In rats given 2 and 20 mg/kg thorium dioxide intratracheally (first and second groups) the counts of the peripheral blood cells were not outside normal limits for a long period. Only in individual animals of these groups was it possible, 10-14 months after intratracheal administration, to note transient anaemia accompanied by considerable reticulocytosis. Marked change in erythropoiesis was noted only in rats given high doses of thorium dioxide (300-400 mg/kg). In the animals of these groups (third and fourth), starting from 6 weeks and especially 3 and 6 months after administration of T h 0 , a considerable 2 number of cases of marked reticulopenia were observed with a reticulocyte count at about 0-9 per cent and lower. Thus, changes in erythropoiesis are related to dose. Some depression of leucopoiesis was noted in rats of the third group in the 7th and 9th months and in animals of the fourth group in the 4th month. Cytological changes in the leucocytes were seen before numerical ones. These changes (fragmentation of the nuclei of the neutrophils, hyper- segmentation and chromatinolysis) were of the degenerative-destructive character, but also were possibly associated with disturbances in the pro- cesses of maturation of the neutrophils. The number of lymphocytes fell considerably after 7-8 months in rats given 200 mg/kg thorium dioxide and in rats of the fourth group (300 mg/kg), 3 months after administration. Together with the numerical, morphological changes were seen in vacuolization of the protoplasm, and sometimes also of the nuclei of the lymphocytes. At later stages, binucle- ated lymphocytes appeared as a result of amitotic cell division. Changes in thrombocytopoiesis reflected in thrombocytosis appeared only in rats which had received 200 mg/kg and more thorium dioxide. Thus, at late stages after intratracheal administration of thorium dioxide changes in erythropoiesis and leucopoiesis appeared similar to those 4 The Toxicology of Radioactive Substances observed as a result of oral administration of radioactive isotopes (Belo- borodova and Baranova, 1957; Beloborodova, 1960; Beloborodova, Ponomareva and Red'kina, 1962; Sagaidak et aL, 1962; and others). The character of these changes and the late times of their appearance allow us to relate them to the radioactivity of thorium dioxide. A characteristic late effect of the biological action of thorium dioxide according to Yan Syao-Shan is seen in the fall in arterial pressure in rats given thorium dioxide via the intratracheal and intraperitonal routes. No distinct relation was established between the fall in arterial pressure and the dose of thorium given. The difference was only in the times of appearance of hypotension. Thus, in the rats which received from 2 to 200 mg/kg thorium dioxide it appeared in the period from 11 to 12 months and for higher doses (300, 400 and 1000 mg/kg) 6-7 months after administration. It should be noted that after intraperitoneal injection of high doses of thorium dioxide the fall in blood pressure set in later than for intratracheal administration. Possibly this is connected with the huge receptor surface of the lungs, irritation of which causes considerable disturbances in the regulation of blood circulation. Such a reaction of the vascular system appearing at late stages after administration of thorium dioxide may also be related to the radiation effect of thorium since from the numerous ex- perimental and clinical findings it is known that ionizing radiations produce hypotension. The function of the central nervous system was quite stable inasmuch as it can be judged from the threshold of stimulation of the neuromuscular apparatus of a hind paw of the rat. During 18 months' observation no sig- nificant changes were observed which could be linked with the action of thorium dioxide. It is possible that use of more refined methods might reveal changes in the functional state of the central nervous system. Nev- ertheless, these findings suggest absence of organic lesions of the central nervous system in the conditions of our experiment as indicated by the results of pathomorphological investigation. Of great interest are the results of the morphological investigations car- ried out by Gaidova and Yan Syao-Shan. It was found that a single admin- istration of thorium dioxide produces histological lesions in the lungs and the parenchymatous organs, directly related to the dose of substance administered. The most serious lesions were observed in the lungs. They appeared on intratracheal injection of thorium dioxide and were of a uni- form nature. They were manifest in the development of suppurative pro- cesses in the bronchi, with sclerotic changes in the peribronchial tissue, development of pneumonia, abscesses and adenomatous proliferations. At late stages, after 13-21 months, 13-6 per cent of the test rats had devel- oped tumours which proved to be sarcomas or squamous cell cancer of Toxicology of Insoluble Compounds of Thorium-232 and Uranium-238 5 the lungs. The minimum dose at which the first tumours were seen was 5 2 mg/kg body weight which corresponds to an activity of 19*2 X l0~ μ Ο per kg body weight. Histoautography revealed tracks of the path of alpha-particles and stel- late figures in the lung tissues of these rats 13-18 months after adminis- tration of thorium dioxide. All this suggests that thorium dioxide possesses low true toxicity so that even for high doses (intratracheal and intraperitoneal) no marked changes are found in the early stages after administration. The late biological effects such as changes in the peripheral blood, reduction in blood pressure and appearance of neoplasms corresponds to those produced by internal and external irradiation with different ionizing radiation sources. All this and also the fact that the neoplasms appear on intratracheal administration of low doses exceeding only ten times the maximum per- missible ones for single administration of thorium dioxide places it in the category of one of the most toxic radioactive substances, which is also con- firmed by the considerable experience with the clinical diagnostic use of colloidal thorium dioxide (Thorotrast). In the present collection we have considered it pertinent to include a description of the technique of determination of thorium in biological media devised by Pavlovskaya and Cherkashina, since the high sensitiv- ity of this technique made it possible to obtain reliable quantitative data on the thorium contents of various biological media. This technique may be useful for those working with thorium. The review also contains laboratory studies on the acute and chronic effects of the very poorly soluble uranium compound U3O8. These inves- tigations were carried out in 1947-50 and prompted work in the radio- toxicology laboratory on the comparative effects of soluble and insoluble compounds of radioactive substances among which a place must be assigned to uranium. The published findings on toxicity of uranous-uranic oxide ( U 0 ) are 3 8 few. The work of Haven and Hodge (1949) and Dygert et al. (1951) by no means exhausted all the problems of the toxicology of this compound and the observation period was merely a month. The investigations of Rubanovskaya showed that negligible amounts of U3O8 are absorbed in the gastro-intestinal tract of the order of hun- dredths and thousandths of a per cent of the amount given judged by the urinary excretion of uranium and its content in the kidneys. Consequently, dogs given by mouth a total dose of 17-100 g of this compound over a long time remained practically healthy and U 0 was almost completely 3 8 removed via the gastro-intestinal tract. Determinable amounts of uranium were found only in the kidneys and thyroid. 6 The Toxicology of Radioactive Substances As shown by the investigations of Beloborodova in these dogs no changes in the red blood cells were found. The white cells tended to show neu- trophilesis. The absolute numbers of lymphocytes and monocytes rose. The morphological investigations of Tolgskaya revealed some hyperplasia of the reticuloendothelial cells of the spleen and lymph nodes. In the kidneys a minor degenerative process was observed and local circulation disturbances in the walls of the intestines. The results suggest that U O has low but marked toxicity which despite 3 s the insignificant absorption of this compound in the gastro-intestinal tract is manifest at different times of treatment. Unfortunately, our inves- tigations were confined to observations for one year which prevented us from following up these animals at late stages when they might have shown a radiation effect from the U 0 given. 3 8 The findings on intratracheal injection of U O to rabbits and dogs are 3 g of interest. In these conditions of administration uranium was found for a long time in the lungs. The investigations of Rubanovskaya showed that over a long period the lungs are cleared of particles of U 0 . For example, 3 8 4-9 days after intratracheal administration from 53-79 per cent of the amount of U 0 administered was found in the lungs of rabbits. Similar 3 8 amounts of thorium dioxide were found in rats at the same times after intratracheal administration. After 2-5 months from 15-53 per cent U O 3 s was detected in the rabbits and after 5-5 to 10 months from 5-5 to 35-6 per cent of the amount given (about 10-50 per cent of the amount found in the lungs in the first days). Thus, during 10 months' observation uranium was removed from the lungs. The latter was found only in small amounts in the urine and faeces. It is possible that such vigorous excretion of uranium from the lungs is connected with the fact that, as shown by the investigations of Tolgskaya, large accumulations of uranium granules are freely situated in the lumens of the alveoli and in the interalveolar septa. The reaction of the surround- ing connective tissue is weakly marked and no nodular formations of a connective tissue character appear as found, for example, for quartz dust. The absence of a marked fibrous reaction is possibly connected with the radioactivity of uranium. The appearance in the pulmonary tissue of foreign body cells around the accumulations of uranium is also charac- teristic. The catarrhal-desquamative inflammatory process in the lungs is highly marked. Intratracheal administration of considerable amounts of U 0 (up to 3 8 1 g) in dogs causes acute poisoning similar to that described by Woroschil- sky in 1889 for the soluble uranium salts. These animals 10-12 days after administration of U 0 develop severe lesions, chiefly of the kidneys, which 3 8 we do not observe on oral administration even of high amounts (up to Toxicology of Insoluble Compounds of Thorium-232 and Uranium-238 1 100 g). These findings show that in relation to the chemical toxicity, intra- tracheal injection of insoluble uranium compounds presents considerable hazards. Tolgskaya presents a detailed morphological description of acute and chronic involvement in rabbits and dogs on oral and intratracheal admin- istration of U3O8. The main changes were found by her in the lungs, kid- neys, the subcortical nodes of the thalamo-hypothalamic region and in the medulla oblongata with insignificant changes in the cerebral cortex and other organs. The investigations of Kurlyandskaya were concerned with the perme- ability of the placenta for U 0 , the possibility of its excretion by the 3 8 mammary glands and also the influence of this compound on intra-uterine foetal development and in the early post-natal period. These investigations showed that following oral and intratracheal administration of the virtu- ally insoluble uranium compound to the pregnant animal, uranium is found in the placenta and foetal tissues. It is also found in the milk of lactating animals and in the tissues and urine of puppies and rabbits fed with the milk of females burdened with uranium. Prolonged observation of the state and development of animals ex- posed to the chronic action of U 0 and their progeny established that it 3 8 exerts a certain effect on the reproductive ability of the females and on the uterine and early post-natal development of rabbits and dogs. The findings obtained concur with those described by Maynard and Hodge (1949) who found a reduction in the litter size for rats exposed to the chronic action of uranium nitrate. Our laboratory findings on the toxicology of U O do not diverge from 3 s those of Dygert et al. (1951) who showed that inhalation of U 0 in con- 3 8 3 siderable concentrations (20 mg/m ) may cause heavy damage to the kid- neys and even death. On the basis of the results we may judge chiefly the chemical toxicity of U 0 since the observation times, as stated above, were not long enough 3 8 to detect radiation damage in rabbits and dogs. This is due to the low specific activity of uranium and the consequent need for a longer time for the onset of the late sequelae as occurs in rats following the intratracheal administration of thorium dioxide. The short review given in the present paper far from exhausts all the aspects of the toxicology of the oxide of thorium-232 and uranous-uranic 238-uranium, which are detailed in the papers in the present collection. Its purpose was to give some generalizations and sharpen awareness of certain patterns important for the understanding of the toxicology of these substances. 8 The Toxicology of Radioactive Substances References 8 9 BELOBORODOVA N. L. and BARANOVA Y E . R, Radiotoxic Effect of Strontium under the Conditions of a Continuous Experiment (Radiotoksicheskoye deistviye radio- aktivnogo strontsiya ν usloviyakh khronicheskogo eksperimenta). In: Toxicology of Radioactive Substances (Materialy po toksikologii radioaktivnykh veshchesty), Vol. 1, p. 151. Moscow (1957). BELOBORODOVA N. L. and BARANOVA Y E . F., Changes in the Peripheral Blood and Cer- tain Other Factors on Continuous Administration of Radioactive Caesium (Izme- neniya perifericheskoi krovi i nekotorykh drugikh pokazatelei pri khronicheskom vozdeistvii radioaktivnogo tseziya). Ibid., p. 162. BELOBORODOVA N. L. and BARANOVA Y E . F., Effect on Haemopoiesis of Prolonged Oral Administration of Radioactive Ruthenium (Vliyaniye na krovotvoreniye dlitel'- nogo wedeniya cherez rot radioaktivnogo ruteniya). Ibid., p. 166. BELOBORODOVA N . L. and BARANOVA YE. F., Investigation of the Functional State of the Haemopoietic System in Rabbits subjected to Continuous Administration of Radioactive Caesium, Strontium and Ruthenium (Issledovaniye funktsionaPnogo sostoyaniya krovotvornoi sistemy u krolikov, podvergavshikhaya khronicheskomu vozdeistviyu radioaktivnogo tseziya, stronttsiya i ruteniya). Ibid., p. 171. BELOBORODOVA N. L., Changes in Haemopoiesis during Prolonged Internal Adminis- e o tration of C o (Izmeneniye protsessov krovotvoreniya pri dliternom wedenii 6 0 vnutr' Co ) . In: Toxicology of Radioactive Substances, Vol. 2, p. 39. Moscow (1960). BELOBORODOVA N. L., PONOMAREVA V. L. and RED'KINA Ε. K., The Effect of Prolonged 5 9 F e Administration on Haemopoiesis (Krovotvoreniye pri khronicheskom deistvii 5 9 Fe ) . In: Toxicology of Radioactive Substances, Vol. 3, p. 48. Moscow (1962). DYGERT H. P., LABELLE C. W . , LASKIN S., POZZANI U. C , ROBERTS E., ROTHERMEL J. J., ROTHSTEIN Α., SPIEGL C. J., SPRAGUE G. F. and STOCKINGER Η. E., Toxicity fol- lowing Inhalation (Toksichnost' uranovykh soyedinenii pri ingalyassionnom w e - denii). In: The Pharmacology and Toxicology of Uranium Compounds, Vol. 1, p. 423. Foreign Literature Publishing House, Moscow (1951). HAVEN F. and HODGE H. C , Toxicity following Parenteral Administration of Certain Soluble Uranium Salts. In: Pharmacology and Toxicology of Uranium Compounds, Vol. 1, p. 281. McGraw-Hill, New York (1949). MAYNARD E. A. and HODGE H. C , Studies of Toxicity of Various Uranium Compounds when fed to Experimental Animals. In: Pharmacology and Toxicology of Uranium Compounds, Vol. 1, p. 309. McGraw-Hill, New York (1949). MCCLINTON L. T. and SCHUBERT J., The Toxicity of Some Zirconium and Thorium Salts in Rats. / . Pharm. Exp. Ther. 9 4 , 1 (1948). PONOMAREVA V. L., Changes in Erythropoiesis during Prolonged Administration of 5 9 5 9 F e (K voprosu ob izmeneniyakh eritropoeza pri dliternom vozdeistvii Fe ) . In: Toxicology of Radioactive Substances, Vol. 3, p. 56. Moscow (1962). POZZANI J., Vysokokachestvennaya ruda (High-grade Ore). In: The Pharmacology and Toxicology of Uranium Compounds, Vol. 2, p. 115. Foreign Literature Publishing House, Moscow (1951). SAGAIDAK N. D. , Peripheral Blood Changes in White Rats following Intratracheal 5 9 Injection of Various F e Compounds (Izmeneniya perifericheskoi krovi pri intra- 5 9 trakheaPnom wedenii belykh krysam razlichnykh soyedinenii Fe ) . In: Toxicology of Radioactive Substances, Vol. 3, p. 86. Moscow (1962). WATANABE S., Pharmacological and Toxicological Studies on Thorium, a Rare Earth Metal. / . Tokyo Med. College 1 5 , No. 2, 121-439 (1957). WOROSCHILSKY L., Wirkung des Urans. Diss., Dorpat (1889).