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(' , ' ^TOJT'COLOGY RESEARCH LABORATORY n 1 TTtO r ^ FP&S 1160-070 i/jVii.'p DOW CHEMICAL U.S.a. OAT May 11, 1976 linin HET K-28126 -fU) num|CM k K-28126 EFFECTS FROM CHRONIC INHALATION OF ASBESTOS PIPE-COVERING DUST IN RATS AND HAMSTERS . REPORTED BY: L. E IHrOKMtriVC SUWMAM HCPCaTCO cxpojumc m Pa-lC J. Leong, R. J. Nyciba, Checked Jv. Frauson, R^,W. Lisowe, L By: L. W. Ramo -- w n COHCLUSlONI lAJCO O M TH| lAM^LC *IC(iV(0. AOO'Y'0*AL iNfONMAfiQH tMC LUOlNO TNI KF^IC TI C *COU'CO A* uSCI ANO fOBMULATlONI AMI OIVILOMO OR \P RROCI1I CHnU CCU*. Rats and hamsters were exposed to the dust of pulverized asbestos pipe covering at an average concentration of 85 mg/M^ for 6 hours per day, 5 days per week for 7 months followed by a lifetime observation period. . In rats, the pulmonary responses were alveolar adenomatous proliferation, non-progressive fibrosis, squamous metaplasia and a substantial incidence of pulmonary carcinoma formation. --< O O jr ^2 A smaller group of hamsters exposed under these conditions experienced an earlier onset of mortality than control hamsters not subjected to the exposure regimen. Although this prevented conclusive evaluation of the pulmonary re sponse in this species, there were no pulmonary neoplasms noted in'the surviving hamsters. See cc < ion on back of re: Vi tf <*/ STOO12461 \o o is EFFECTS FROM CHRONIC INHALATION OF ASBESTOS PIPE-COVERING DUST IN RATS AND HAMSTERS B. K. J. Leong, R. J. Kociba, L. E. Frauson, R. W. Lisowe and L. W. Rampy Toxicology Research Laboratory Health and Environmental Research Dow Chemical U.S.A. Midland, Michigan 48640 May 11, 1976 ST0012462 STOO I 2.1,63 ABSTRACT Rata and hamsters were exposed to the dust of pulverized asbestos pipe covering at an average concentration of 85 mg/M^ for 6 hours per day, 5 days per week for 7 months followed by a lifetime observation period. In rats, the pulmonary responses were alveolar adenomatous proliferation, non-progressive fibrosis, squamous metaplasia and a substantial incidence of pulmonary carcinoma formation. A smaller group of hamsters exposed under these conditions experienced an earlier onset of mortality than control hamsters not subjected to the exposure regimen. Although this prevented conclusive evaluation of the pulmonary re sponse in this species, there were no pulmonary neoplasms noted in the surviving hamsters. STOO12463 W 2 f 001S 2 INTRODUCTION Asbestos materials, primarily chrysotile, crocidolite, and amosite, are used extensively as thermal insulation mater ials.^" Many industrial products worldwide rely increasingly on asbestos and the variety of its uses. At the same time, asbestos is also being perceived as a major health hazard. Asbestos dust exposure is now widely recognized as an important etiological factor in pulmonary fibrosis, 2 ' 3 ' 4 res piratory cancers,^ ^ and malignant mesothelial tumors.^ ^ However, the precise mechanisms of the causal relationships have not yet been clarified although many theories have been proposed. Several factors in the carcinogenic response to asbestos have been identified: (1) the action of the fiber it self due to its particular geometry,^(2) trace metals associated with the fiber (some of which are known to be carcinogenic themselves), ' (3) polycyclic aromatic hydro carbons associated with the fiber,^ (4) other carcinogenic agents adsorbed onto the asbestos during milling, handling, mining, etc. and carried on the asbestos fiber into the body, 19 and (5) cigarette smoke in conjunction with asbestos exposure."^ Considering these factors, the theories basically view asbestos fiber as a carcinogen or as a carrier of a car cinogen. . STOO12464 STOOI 2 U65 3 Most of the asbestos inhalation experiments to date were conducted using relatively pure minerological types of dust. The findings in rat3 and hamsters exposed to asbestos dusts generated from prefabricated asbestos pipe covering are pre sented in this paper. The duration of exposure was 6 hours per day, 5 days per week for 7 months followed by observa tion for the life of the animals. ST0012465 STOOI2U66 4 MATERIAL AND METHODS Material Asbestos dust was prepared by pulverizing a section of a prefabricated asbestos air-cell pipe covering into coarse powder with an industrial hammer mill for 5 minutes. The coarse powders were further reduced to micron and subraicron sizes in a ceramic ball mill running for 48 hours. The final dust_sample consisted of 30-40% of noncrystalline and approximately 60% of crystalline materials as determined by X-ray diffraction analysis. The crystalline components con sisted of approximately 50% asbestos form materials (chrysotile), 5% Fe^O^, 5% Mg(OH)2 and 5% MgO. Atomic emission spectrographic analysis of a representative sample revealed the presence of various minerals as shown in Table 1. Dust Atmosphere Generation The asbestos dust atmosphere was generated by using an air elutriator as shown in Fig. 1. The elutriator consisted of a 90 x 10 cm pvrex tube. The bottom of the tube was sealed with a rubber diaphragm attached to an air vibrator. The powder was poured through the opening on top of the tube onto the rubber diaphragm. The powder was then churned up by the vibrating diaphragm and was carried upward by a swirling air current emerging from two air nozzles located ST0012466 L9lZ I 001S 5 just above the diaphragm. The quantity of fine dust to be introduced into the exposure chamber could be regulated by adjusting the vibration frequency of the diaphragm and the volume of air ejecting through the dust cloud. For a 6 hour exposure period 20 grams of powder were initially placed in the dust generator. Additional 10-gram portions of powder were added twice daily to maintain a fairly constant level of dust in the chamber atmosphere. The coarse residue was discarded at the end of each experimental day. Chamber Dust Concentration Analyses The concentration of asbestos dust in the chamber atmosphere was determined gravimetrically using a membrane filter-sampler technique. Two open-face dust samplers were placed in the center of the exposure chamber just above the animal holding cages. A third sampler was placed within an empty cage sur rounded by other cages containing animals. An air sample was drawn at the rate of 8.5 liters per minute for 5 to 10 minutes through a 47 mm diameter membrane filter (Metricel GA-4) of 0.8 um pore size. The asbestos dust retained by the filter was weighed so that the dust concentration, in terms of milli grams per cubic meter of air, could be determined. ST0012467 8972 I 001S 6 Particle Size Distribution Analyses The particle size distribution of the airborne asbestos dust in the chamber was monitored using a light scattering particle counter. Dust in air samples drawn from the center of the chamber was analyzed for particle size distribution. The dust trapped in a membrane filter sampler was dispersed in a suitable mounting medium and counted automatically with a phase contrast microscope interfaced with an automatic scan ning image analyzer. The digital outputs from the particle analyzer or image analyzer were computerized and the numberlength-mean diameter of each sample was calculated. The dusts trapped.in the membrane filter sampler were subjected to scanning electron microscopy to determine the particulate and . fiber content of the sample. Experimental Design 50 male Sprague-Dawley Specific Pathogen Free (SPF) derived rats of Spartan substrain and 15 male Golden Syrian hamsters were exposed to the asbestos dust. The duration of exposure was 6 hours per day, 5 days per week for a total of 150 exposures in 7 months. The exposures were conducted in a one-cubic meter stainless steel and glass chamber under dynamic airflow conditions. During the exposure, the animals were housed individually in wire gauze cylinders 3 inches in diameter and 12 inches long. The cylinders could be piled ST0012468 ST00I2469 7 together up to 5 layers deep without affecting the dis tribution of dust to the animals. At the end of each ex posure day, the animals were removed from the cylinders and housed in gang cages with food and water. A comparable group of 50 rats and 15 hamsters was maintained in an animal holding room to serve as controls. These animals were not placed in individual cylinders daily but were deprived of food and water on a schedule identical to that of the exposed animals. At the termination of the 7-month exposure period, the ex posed animals were housed in a holding room with the control animals. Animal Observation' During the exposure period all animals were observed for evidence of ocular, nasal and respiratory irritation. Body weights were determined once a week for approximately 8 months (7 exposure months and 1 month post-exposure) and monthly thereafter until all animals died. The body weight data were analyzed by analysis of variance and Dunnett's test, p<0.05. Pathology p Interim kills of 5 control rats and 5 rats exposed to as bestos were conducted at the end of the 7-month exposure. ST0012469 STOO 1247 0 8 and at 1 and 6 months post-exposure (8 and 13 experimental months). A similar interim kill at 18 months post-exposure (25 months of the study) was conducted on 4 control and 4 exposed rats. (Hamsters were not killed at any of the interim kills because of the limited group size). At these intervals, the designated rats were starved over night and killed by decapitation. Immediately prior to sacrifice, the rats' tracheas were clamped under methoxyflurane anesthesia in order to avoid aspiration of blood. The lungs and trachea were removed and subsequently dis tended with phosphate buffered 10% formalin. The lungs, thoracic lymph nodes and representative portions of heart, kidney, liver, spleen, adrenal, testes, trachea, esophagus, accessory sex glands, stomach, thyroid, parathyroid, aorta, pancreas, small intestines, brain, pituitary, urinary bladder and any other tissues suggestive of a pathologic process were preserved in formalin fixative. Nasal turbinates from rats of the 6 month post exposure (13th month of the study) kill were also preserved in the fixative. Routine pro cedures were used to prepare paraffin-embedded tissue sec tions that were subsequently stained with hematoxylin and eosin (H&E). Additional sections of lungs and thoracic lymph nodes from all interim-kill rats were stained with special stains to detect the presence or absence of collagen ST0012470 STOOI 247 I 9 (Masson's Trichrome stain) and of reticulin (Gordon and Sweet's, or Gridley's or Gomori's staining procedure). The resultant sections of all rats were examined for histo pathologic changes. Special attention was given to thelungs and thoracic lymph nodes to define the response to repeated inhalation of the asbestos dust. All rats not killed at interim kill dates were maintained under control conditions until they died spontaneously or were killed in a moribund condition (maximum of 870 days after start of exposure). All hamsters were retained under control conditions until they died spontaneously or were killed in a moribund condition (maximum of 723 days after start of exposure). All these animals were subjected to necropsy examination and the lungs were distended with phosphate buffered 10% formalin. The lungs, thoracic lymph nodes, and representative portions of heart, kidney, liver, spleen, adrenal, testes, trachea, esophagus, and any other tissues suggestive of a pathologic process or tumor formation were preserved in formalin fixative. Hematoxylin and eosin stained sections of these tissues were routinely prepared for histopathologic examination. Thorough gross and histopathologic examination was conducted on tissues of rats and hamsters in order to (a) ascertain the probable cause of death, (b) define possible pulmonary response, and (c) evaluate tumor incidence. ST0012471 ZU721 001S 10 Tumor incidence data were statistically evaluated with Fisher's Exact Probability Test. In all cases, probability values (p) of less than 0.05 were interpreted as indicating significant differences. ST0012472 STOO I 21,73 11 RESULTS Chamber Atmosphere A typical sample of asbestos dust retained on a membrane filter is shown in the scanning electron micrograph (Fig. 2). The dust consisted mainly of amorphous particulate matter and some fibrous material. Gravimetric analyses of over 400 samples of asbestos dust indicated that the average dust concentration in the chamber atmosphere was 85 f 52 milligrams per cubic meter of air. Particle size distribution analyses of the dust samples dis persed in mounting medium revealed an average number-lengthmean diameter of 0.8 urn. However, direct measurement of the dust particles suspended in air with a light scattering par ticle counter showed an average number-length-mean diameter of 4.1 ym. The larger diameter recorded by the light scat tering measurement technique suggests agglomeration of dust particles suspended in the chamber atmosphere. Animal Observation During each exposure period, signs of slight ocular and nasal irritation were observed in both rats and hamsters. The animals buried their muzcles in the hair coat. The animals remained inactive until they were removed from the ST0012473 STOOI 2474 12 dust atmosphere at the termination of each exposure. The body weight data (Fig. 3) indicated that after 1 month of exposure to asbestos dust, the mean body weights of rats were significantly lower than those of control rats. Weight gain improved slightly following termination of the exposures, but the mean body weights remained significantly lower than those of the control group until 14 months post exposure (21 experimental months). The mean body weights of the exposed hamsters were statistically decreased from those of the control animals on two occasions during the first month of the experiment. After 3 months of exposure, the test ham ster's body weights remained statistically increased com pared to controls, except on isolated occasions, until the end of the study. Mortality data for rats (Fig. 4) indicated no significant difference between exposed and control rats. Ninety percent of the rats died during the observation period between the 7th and 30th experimental months. Forty percent of the hamsters exposed to asbestos dust (Fig. 5), died during the 7 month exposure period. Remaining exposed hamsters died within 10 months post-exposure. Control hamsters did not start dying until 2 months post-exposure and some survived as long as 16 months post-exposure (23 experimental months)'. Thus, the lifespan of hamsters appeared to be significantly reduced by the exposure regimen used in this study. ST0012474 ST00 I 2U75 13 Pathology - Rats (See Tables 2 & 3*) In rats killed at the end of the 7-month exposure, pul monary deposits of the dust were visible grossly as isolated subpleural pale foci, usually less than 2 nun in diameter. Microscopically, the principal changes were isolated aggre gates of dust-laden macrophages within alveoli. Many of the affected alveoli were those that evaginate off the alveolar ducts or repiratory bronchioles of the pulmonary raceme. The alveolar membranes adjacent to these aggregates showed a very slight focal hypercellularity and thickening, but no discernible increase in reticulin or collagen deposition. The pulmonary interstitium and satellite thoracic lymph nodes sometimes contained aggregates of the dust, both extra cellular and intracellular within macrophages, with typically no significant increase in reticulin or collagen content. At subsequent evaluations during the post-exposure period (8, 13 and 25 months of the experiment), pulmonary reaction similar to that described at 7 months was noted. There were no marked accumulations of intraalveolar proteinaceous material, leukocytes or inflammatory debris nor were there marked fibrcgenic responses in the lungs. Appendixes A-D to this report contain gross and histopathologic observations on control and asbestos-exposed rats dying during the course of the study and also gross and histopathologic observations or. non-puimonary tissues examined at the interim examinations. ST0012475 STOOI2U76 14 During the period from the 14th month to the end of the study at 29 months, the pulmonary response to asbestos also included some cases of focal alveolar hyperplasia, alveolar adenomatous proliferation, non-progressive fibrosis and squamous metaplasia of alveoli. Pulmonary deposits of asbestos dust were no longer grossly visible by this time. More importantly, 6/34* rats exposed to asbestos and ex amined after months 13 of the study had pulmonary carcinoma formation as compared to 0/35* control rats with pulmonary tumors. The pulmonary carcinomas were seen in rats that died from 16 to 29 months after the initiation of the ex periment (294 to 571 days post-exposure). The incidences of all types of tumors found in control and asbestos exposed rats are summarized in Table 4**. Statistical evaluation using Fisher's Exact Probability test showed that the incidence of pulmonary carcinomas was significantly increased in the group exposed to asbestos. The incidences of non-pulmonary tumors were comparable in both asbestos exposed and control rats. Histopathologic studies of other, non-puimcnary, tissues revealed no alterations considered related to exposure to the asbestos dust. * A total of 15 rats per group had been killed for interim examir.attens at 7, 3 and 13 months cf the study and one ex posed rat was missing. Appendixes E and E contain a listing of the temporal in cidence of all tumors in control and asbestos exposed rats respect iveiy. ST0012476 STOOI 2477 15 Pathology - Hamsters* Hamsters that died during the exposure period had evidence of ulcerative enteritis and/or amyloidosis. Since ulcerative enteritis and amyloidosis can occur spontaneously in hamsters and were, in fact, later found in several control hamsters, it is difficult to consider the exposure to asbestos as the sole factor in this early mortality. However, it appears as if the exposure to asbestos or the stress of the exposure regimen was at least partially re sponsible for the earlier mortality observed in these hamsters exposed to asbestos. Two hamsters died during the exposure period with pulmonary lesions, one with focal consolidation of the lungs and one with pulmonary ectatic emphysema. These isolated cases may or may not have been related to asbestos dust exposure. All other tissue alterations in hamsters were considered spon taneous in nature and unrelated to treatment. The ability to evaluate long-term pulmonary effects associ- 1 ated with chronic inhalation of the asbestos was severely limited by the early mortality of 8/15 hamsters. The fol lowing comments are based on the examination of the remaining Appendixes G and H to this report contain mortality data and gross and histopathologic observations on individual control and ashestos-exposted hamsters respectively. STOO12477 ST00I 2478 16 7/15 hamsters that died spontaneously from 10-16 months after initiation of the study. Deposits of the asbestos dust were not grossly visible in the lungs or thoracic lymph nodes. Microscopically, the principal pulmonary changes in exposed animals were isolated aggregates of particle-laden macrophages within alveoli. The alveolar walls adjacent to these aggregates were slightly thickened. Minimal focal alveolar epithelial hyperplasia near the affected bronchioles of the lung was observed in 3 of the 15 hamsters examined. In the control group of ham sters major pulmonary lesions were as follows: focal alveolar epithelial hyperplasia in 2/15 hamsters; eosinophilic material and pigment-laden macrophages in alveoli of 2/15 hamsters; cholesterol clefts and mineralized deposits in 1/15 hamsters. Based on the equivocal observations made on this limited number of hamsters, there was no conclusive evidence of a significant pulmonary response to exposure to asbestos dust in this species. There were two cases of tumor formation in control hamsters, one a malignant lymphoma with widespread metastasis and one an adrenal cortical adenoma. There were no tumors observed in the 15 hamsters exposed to asbestos. This observation must be considered in light of the fact that 6 of the 15 ST0012478 S T 0 0 I2 4 7 9 17 hamsters in the asbestos group died within the initial 7 months of the study, whereas no controls died before the 9th month of the study. ST0012479 STOOI2U80 la DISCUSSION The results of the present study indicate that exposure to asbestos pipe-covering dust at an average concentration of 85 mg/M^, for 6 hours per day, 5 days per week for 7 months caused pulmonary carcinomas in SPF-derived Sprague-Dawley male rats. The earliest pulmonary carcinoma was found in a rat that died 477 days (approximately 16 months) after initi ation of exposure. Pulmonary carcinomas were found in ^18% of the rats surviving longer than 13 months. In studies by Gross et aJL. 21 (1967), rats which were not SPF derived were exposed to chrysotile fibrous dusts. The average dust concentration was 86 mg/M^ and the duration of exposure was 62 weeks (approximately 14 months). The inci dence of tumors observed in their rats was approximately 16% and the first pulmonary carcinoma was found in a rat that died 16 months after initiation of exposure. In a comparison of the results of the two studies, it is interesting to note that the incidence of tumors in their rats was approximately the same as in our rats even through their animals were ex posed for twice as long to a similar concentration. The latent period for tumor development was practically the same (16 months) in both studies. 22 In similar studies by Reeves et a_l. (1971), rats w" ere ex posed to chrysotile dust at a lower concentration of 48 mg/'M^ STOO12480 STOOI 248 I 19 for only 4 hours a day, 4 days per week for 99 weeks (ap proximately 23 months). No pulmonary cancers were found. However, in an identical inhalation study with crocidolite asbestos dust, Reeves et a_l. noted a low incidence of pul monary squamous-cell carcinomas. Two of the factors believed to be associated with carcino genicity of asbestos are (1) the morphology and (2) the trace metal contents of the material. Morphologically, the as bestos dusts used in the present experiment contained mainly 0.5 to 10.5 u particles and very few fibers (Fig. 2). Thus, the production of pulmonary carcinoma in the present experi ment supports the conclusion of Kogan e_t al_. , who considered fiber length or fibrous structure as non-essential for patho genicity.23 Regarding the trace metal contents (Table 5), the quantities of nickel, chromium and cobalt in the asbestos sample used in our experiment were quite comparable to those of the UICC (Union Internationale Contre Le Cancer) reference standard sample, 24 but were 2 to 4 times higher than the chrvsctile samples of Gross et al_. , 21 and Reeves et al. 22 The high content of trace metals in the sample used in the present experiment may be an explanation of its high carcino genic potency. On the other hand, the success of producing ,lung cancer with crocidolite 2 2 which contained rather low concentrations of nickel and cobalt indicates .that the ST0012481 STOO I 2482 20 f mechanism of pulmonary carcinoma induction by asbestos dusts may not be entirely related to the content of trace metals. Additional studies are needed to elucidate the mechanism of carcinogenesis by asbestos dust. Furthermore, it would be necessary to examine the chronic effects from inhalation exposure to lower dust concentrations to determine a real istic industrial handling guideline based on animal studies. ST0012482 0012483 REFERENCES Hendry, N. W. 7he Geology, Occurrences, and Major Uses of Asbestos. Ann, New York Acad. Sci. 132:12-22, 1965. Cooke, W. E. Pulmonary Asbestosis. Br. Med, J. 2:1024 1025, 1927. Merewether, E. R. Asbestosis and Carcinoma of the Lung, in Annual Report of the Chief Inspector of Factories for the Year 1947. London, His Majesty's Stationery Office, 1949, pp 79-91. Murphy, R. L., Ferris, B. G., Burgess, W. A., et al. Effects of Low Concentrations of Asbestos. N. Engl. J. Med. 285:1271-1278, 1971. ' Selikoff, I. J., Churg, J. and Hammond, E. C. The Occurrence of Asbestosis Among Insulation Workers in The United States. Ann. New York Acad. Sci. 132:139-153, 1965 . Hammond, E. C., Selikoff, I. J. and Churg, J. Neoplasia Among Insulation Workers in the United States with Special Reference to Intra-Abdcmina1 Neoplasia. Ann. New York Acad. Sci. 132:519-525, 1965. ST0012483 > STOgI 2484 7. Selikoff, I. J., et al. Mortality Experiences of Asbes Insulation Workers. In International Conference on . Pneumoconiosis, Johannesburg, pp. 95-103, 1969. 8. Shettigara, P. T., Morgan, R. W. Asbestos, Smoking and Laryngeal Carcinoma. Arch. Environ. Health 30:517-519, 1975. 9. ' Selikoff, I. J., Hammond, E. C., Churg, J. Asbestos Exposure, Smoking and Meoplasia. JAMA 204:106-112, 1968. . 10. Cooper, W. C. Asbestos as a Hazard to Health. Arch Environ. Health 15:295-290, 1967. 11. Selikoff, I. J., Churg, J., Hammond, E. C. Relationship . Between Exposure to Asbestos and Mesothelioma. N. Enel, J. Med. 272:560-565, 1965. 12. Newhouse, J. L., Thompson, H. Mesothelioma of Pleura and Peritoneum Following Exposure to Asbestos in the London Area. Br. J. Ind. Med. 22:261-269, 1965. 13. Smith, W. E-, Miller, L., Churg, J., and Selikoff, I. J. Mesotheliomas in Hamsters Following Intrapleural Injection of Asbestos, J. Mt. Sinai Hosp. 32:1-8, 1965. 14. Wagner, J. C. and Berry, G. Mesotheliomas in Rats Following Inoculation with Asbestos. Brit. J. Cancer 23:567-581, 1969. ST0012484 STOOI 2 485 15. Berry, G. and Wagner, J. C. Application of a Mathematical Model Describing the Time of Occurrence of Mesotheliomas in Rats Following Inoculation with Asbestos. Brit. J. Cancer 23:582-536, 196~9. 16. Stanton, M. F. and Wrench, C. Mechanisms of Mesothelioma Induction with Asbestos and Fibrous Glass. J. Nat. Cancer Inst. 48:797-821, 1972. 17. Gross, P., Harley, R. A. The Locus of Pathogenicity of Asbestos Dust. Arch. Environ. Health 27:240-242, 1973. 13. Cralley, L. J. and Lainhart, W. S. Are Trace Metals Associated with Asbestos Fibers Responsible for the ' Biologic Effects Attributed to Asbestos? J. Occup. Med. 15 (3) :2 6 2-2 6 6 , 1973. 19. Dixon, J. R. et al. The Role of Trace Metals in Chemical Carcinogenesis: Asbestos Cancers. Cancer Res. 30:1068-1074, 1970. 20. Shabad, L. M., et al. Experimental Studies on Asbestos Carcinogenicity. J. Natl. Cancer Inst. 52(4):11751137, 1974. 21. Gross, P. deTreville, R. T. P., Toker, E. B., Kaschak, M. and Babyak, M. A. Experimental Asbestosis. Arch Environ. Health 15:343-355, 1967. ST0012485 STOO12486 22. Reeves, A. L., Puro, H. E., Smith, R. G. and Vorwald, A. J. Experimental Asbestos Carcinogenesis. Environ. Res. 4:496-511, 1971. 23. Kogan, F. M., Troitskie, S. Y. and Udilova, N. N. Data on the Mechanism of the Fibrogenic Action of Asbestos Dust. Gig. Sanit. 31:10-14, 1966. 24. Timbrell, V., Gilson, J. C. and Webster, I. UICC Standard Reference Samples of Asbestos. Int. J. Cancer 3:406-408, 1963. ST0012486 ST00I2487 o GO TABLE 1 ATOMIC EMISSION SPECTROGRAPHIC ANALYSIS OF ELEMENTS PRESENT IN THE BALL MILLED ASBESTOS SAMPLE USED IN INHALATION EXPOSURE OF RATS AND HAMSTERS Element Weicht % of SamDle A1 Ba B Ca Cd Co Cu Cr Fe Mg Mn , Ni Pb Si Sn Sr Na K Ti V Zn 0.008 <0.05 0.4 <0.05 0.004 <0.001 0.07 2.0 11.0 Q .04 0.09 0.01 10.0 <0.01 <0.001 1.0 <2.0 0.5 <0.005 <0.5 ST0012487 soqssqsv O OO <N Xozquco to o E-* OOO O CO CO E<->. a a a co < w x *" m soqsaqsY in Hoo o , U1 < E-I 2 l-t s c 0 *H X o >* a r-i XOJrquoo c-^ u-> Ooo o X <0 > CO 3 u CO C"1 o 3 CO >n CO co 2 Mo XI o e- IX Cfl E- >< < xX CO soqsaqsv in ao 3 o o << U 2 ITS O2 2M as Xozquoo in ooo o > X <N| 3 (J aj JZ X2 DU k-H X X 2X cr. a O3 c <Q E- CO 2 CO OX i-t < so5.ssqs y m m * o o -C E-1 > <X >X (X x c2 CO I--i a oX Xocrquco in oo a 3 o0 Li O' 4Z 2 o C-l i--i E-1 CJ) O 3 OJ OX X< o XX E- X <X X CO < CO oc XX uX t-- M in sz u C 0 c r-l c > u 0 c a r c- a, 3 in 0 Ui 3 c cs X U_i c0 c Eu X a _7 CJ z W c 0 -.H 4J 3 > u CJ in A O > u 3 c 0 zz Cl in m 0 u C f X 3 W a rH >rj a, 3e CJ n <H fN u1 u <--H tn E 3 uo Li 0 u a; 3 u a3 H O U * c-* 21 SZ X cn E in O' >. cn c c3 3o r-( Cu U-t 3 OL m0 aj 3 r. u CJ u c L, 33 c Li 3 TJ a OJ CL c p., OJ 3 T3 TJ TJ U o c CJ .* 3 lx U 30 C a- -C -U ^ 3 C-T 3 r-H V) 0o 73 4J 0 )X c 3 CL. C 'r* u 3 Li 4i 0 Li OJ X) 3 C in 3 T3 CJ J rH 3 J 3 a CO --4 O CD no -C~ co CO STOO12488 TABLE 3 HISTOPATHOLOGIC OBSERVATIONS ON PULMONARY TISSUES OP MALE RATS KILLED AT SERIAL TIME INTERVALS DURING INHALATION STUDY WITH ASBESTOS ro CO V.O abesl Time Interval in Months 7 Treat--ent Group______________________________ Number of Rats_______________________________ Microscopic Pulmonary Observations: _wou uC0 5 Alveoli and Alveolar Membrane Isolated focal aggregates of parcicle- laden alveolarmacrophages 0 Isolated focus of inf lacuna tory cells In lungs 0 Isolated aggregates of alveolar macrophages 0 Isolated focus of alveolar epithelial hyperplasia 0 Isolated focus of hyperceilulanty of alveolar wall 0 Isolated foci of hyparcellularity and thickening of alveolar walls adjacent to particle-laden alveolar macrophages - very slight to equivo cal .0 Very slight to equivocal increase in collagen and reticulin in affected alveolar walls 0 Very slight to slight increase in collagen and reticulin in affected alveolar walls 0 Pulmonary carcinoma, with both adenomatous and mucinous areas 0 Sarcomatous-1ike proliferation of tissue in lung and thorax 0 Pronounced deposition of connective tissue in lung 0 Adenomatcus proliferation of bronchial epithelium 0 7 wtoavA 1 < 5 5 0 0 0 0 5 3 0 0 0 0 0 8___ a 90} w c n01 ua (A < 55 ___ 13 *ouj c o o 5 13 o71 < 5 05 20 30 00 30 05 03 00 00 00 00 00 05 10 50 00 12 04 00 00 00 00 00 00 Data listed as number of rats in each group showing he various observations 25_____25 ou aO c uo < 44 04 00 00 02 00 04 01 03 01 01 01 01 ST0012489 STOOI 2U90 TABLE 3 (CONTINUED) HISTOPATHOLOGIC OBSERVATIONS ON PULMONARY TISSUES OF MALE RATS KILLED AT SERIAL TIME INTERVALS DURING INHALATION STUDY WITH ASBESTOS Time Interval in Months 7 Treatment Grouo Number of Rats Microscopic Pulnonarv Cbservaticr.a : W u0 *j c 0 u 5 Interstitium Peribronchiolar, perivascular lymphoid aggregates Peribronchiolar lymphoid aggregates containing particle-laden macrophages and particles, with no JigniSicsnc collagen and reticulin 0 0 Thoracic Lymoh Nodes Intracellular and extracellular deposits of particles, within thoracic lymph nodes Submucosal tracheitis,slight 0 0 7 n 0 XI ia V n V) < s 0 1 2 0 88 V) 0 0 *j u9 xt 4> cA 09 u< 55 13 13 n 1-4 0 0 XJ u9 AJ o cA 0 83 'O < 55 20 02 05 00 00 03 03 1I Data listed as number of rats in each group showing the various observations 25 25 91 o o u X* Cl *j o cA 0a < 4. 4 33 01 02 00 STOO12490 I 6*lZ I 001S TABLE 4 SUMMARY* OF TUMOR INCIDENCE IN GROUPS OF MALE RATS USED IN INHALATION STUDY WITH ASBESTOS (Or Used as Controls for the Study) GrouD Control Number of Rats Examined Number of Rats with Tumors Number of Tumors Mean Number of Tumors/Rats with Tumors 50 19 23 1.2 Luna Papillary adenocarcinoma Epidermoid carcinoma Adenomatous and mucinous carcinoma Total Pulmonary Tumors 0/50 0/50 0/50 0/50 (0/35) Pituitary Gland Adenoma 6/50 Adrenal Gland Pheochromocytoma 4/50 Subcutaneous Neurofibroma Carcinoma Fibroma Fibroadenoma 0/50 0/50 3/50 1/50 Pancreas Adenoma Islet ceil adenoma 2/50 1/50 Asbestos 49b 20 34 1.7 3/49 2/49 1/49 6/49* (6/34) 6/49 6/49 1/49 1/49 1/49 0/49 1/49 2/49 aA record of observations on individual rats is contained in Appendixes A and 3. b0ne rat was missing. Q Fifteen of the 49 by the 13th month tumors up to 6 of rats were killed for interim examinations bringing the actual ir.c idence of pulnonar 34. Significantly increased as compared to controls when analyzed by Fisher's Exact Probability Test, p<0.05. STOO12491 STOOI 2492 TABLE 4 (Continued) SUMMARY OF TUMOR INCIDENCE IN GROUPS OF MALE RATS USED IN INHALATION STUDY WITH ASBESTOS (Or Used as Controls for the Study) Grouo Control Number of Rats Examined Number of Rats with Tumors Number of Tumors Mean Number of Tumors/Rats with Tumors 50 19 23 1.2 Brain Craniopharyngioma 1/50 Heart Rhabdomyosarcoma 1/50 Thyroid Adenoma 2/50 Zvmball Gland Carcinoma - 1/50 Small Intestine Mucinous adenocarcinoma 0/50 Testes Interstitial cell tumor 0/50 Intraabdominal Malignant schwannoma Unclassified sarcoma 0/50 1/50 Asbestos 49b 20 34 1.7 0/49 0/49 6/49 0/49 1/49 1/49 1/49 1/49 aA record of observations on individual rats is contained in Appendixes A and B. bOne rat was missing. C F.ifteen of the 49 rats were killed for interim exam'. inati.ons by the 13th month bringing the actual incidence of pulmonary tumors up to 6 or 34. Significantly increased as compared to controls when analyzed by Fisher's Exact Probability Test, p<0.05. STOO12492 cn i4 Ul <4 aa o2 u< to Q 2 < oin E-i X CO u 2u 4 M a so CO < K 2 Du CJ O to 4 U VC E- 2W uM E2- 2O vO ON ON a> ON o u TJ* c CM V o o o ooo uu afi 1 1 1 u 0) a> rvi in HH L-l r* m D 'll cc oo 4o oo o a> rH -D rH o CN 0oo T3 <*> 1o (n <JS QJ CM u O o rH CN 0 o a- H Li flj U o nj cn 4J OJ OJ rH u tn a c OJ rH V> (N V TT O a> o Oo u o uU a: a 0 ooo Uu C2l t/1 00 1 a> CO CO cz Li m cn cn u O4 o O O oo o r-H CN r-H a n3 rH lO m CN jj iJ in CN Q) o <N rH O W V) w >L 0in jC CJ 00 O O o * o O o CJ <JJ 4-> O- in -4 ^ cl c V) CP CTN r- O oc O oo 4-> M 00 in u o oo CJ OJ X3 > in O < CJ 4i~n> c o e CJ e ^H -aH o- 0E o4 4L 4-1 i-H <0 X0) ex CJ o iH T3 O M D QJ pH a E 3 CA in o 4J in a X) in A3 T3 OJ rH rH *H E rH rH XI IM 0 in in >* rH *3 C *3 U -iH JZ (CTJm u CP 0 4J U 0) CL in c 0 in in .^ E CJ u E O 4J ra l4 o cn 31 in a >i T42JJ u in E uOu* . ^in w ST00 I 2493 ST0012493 STOOI 2494 Rubber Stopper To Exposure Chamber Fig. 1. A schematic diagram of the air elutriator dust generator. Compressed air passes flow meters to an air vibrator (A) and two air nozzles (B) to stir up the dust. Air ejected from nozzle (C) blows the dust over to exposure chamber. STOO12494 S612I001S Fig. 2 . Scanning electron micrograph of ashes os dusts re ta i.ned on filter 2i70x (2.5 cm=10u) STOO12495 S T 0 0 I 2 4*9 6 Body Weighis, Grams " Figura 3 MEAN BODY WEIGHTS OF MALE RATS AND HAMSTERS EXPOSED TO ASBESTOS DUSTS OR MAINTAINED AS CONTROLS ST0012496 peaQ % 'uoijeincJOd |eujiuv Figure 4 CUMULATIVE PERCENT M O R T A LIT Y OF MALE RATS EXPOSED TO ASBESTOS DUSTS AND THOSE M A IN TA IN E D AS CONTROLS Experimental Duration, Months OO O o DO -T" lD ST0012497 Figure 5 CUMULATIVE PERCENT M O RTAILITY OF MALE HAMSTERS EXPOSED TO ASBESTOS DUSTS AND THOSE M AIN TAIN ED AS CONTROLS STOO12498 STOOI 2499 APPENDIX A CROSS AND MICROSCOPIC OBSERVATIONS CM RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at tine of Death _____________ Gross and Microscopic Observations 72-105 60 Gross: Killed; moribund, due to traumatic injury to upper incisors and overgrowth of lower incisors; decreased adipose tissue. Microscopic: Odontitis, peridoncitis and gingivitis secondary to crauma; focal interstitial pneumonitis focal hepatocellular degeneration and inflammation. 72-146 109 Gross: Killed; cyst in the region of the brain and pituitary, 1 cm in diameter. Microscopic: Proliferating cystic structure in region of ventral aspect of brain, and pituitary (craniopharyngioma). . 72-134 186 Gross: Killed.; large subcutaneous nodular structure weighing 169.5 grams, consisting of semi-gelatinous tissue with non-fibrous areas; generalized anemia. Microscopic: Subcutaneous fibroma; increased splenic hemopoiesis; increased hemopoiesis in liver. 72-115 409 Gross: Killed; large subcutaneous mass weighing 134.5 grams; heart dilated. Microscopic: Subcutaneous fibroma; slight chronic nephropathy; myocardial degeneration; intestinal nematodiasis. 72-131 409 Gross: Killed; excessive adipose tissue; edema of testes and scrotal region. Microscopic: Vacuolatlon of adrenal cortical cells; pituitary adenoma; focal pancreatic atrophy and fibrosis; hyperplasia of pancreacic islet; slight chronic nephropathy; isolated focus of inflam matory cells in lung. ST0012499 S T00I2500 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE 3EINC MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Sire 50 Male Rats) Pathology Number 72-148 Days on Test at time of Death __________ Gross and Microscopic Observations 413 Cross: Killed; noribur.d; over 130 ml of serosanguineous fluid in abdomen; highly lobulated dense white mass incorporating stomach, small and large intestines, and mesencery; metastatic trans plantations on the diaphragm and peritoneal cavity. Microscopic: Lntraabdominal sarcoma, with extension via abdominal region; moderate chronic nephropathy; increased splenic hemopoiesis; scattering of alveoi macrophages in lungs. 72-149 420 Gross: Died; advanced aucolysis; multiple nodular ' areas in che pancreas 1 mm to cm in diameter; - enlarged kidneys with multiple pale foci on the cortex and dilated renal pelvis; pale circum scribed area on liver; enlarged parathyroid glands. Microscopic: Pancreatic adenoma; pronounced chronic nephropathy; pituitary cyst formation; isolated aggregates of particle-laden macrophages in lung; periarteritis; testicular atrophy; skeletal muscle degeneration; increased splenic hemopoiesis; myocardial degeneration; hepacocellular hyper plastic nodule. 72-139 423 Cross: Killed; enlarged lobes of prostate containing green-yellow exudate. Microscopic: Moderace chronic nephropachy; myocardial degeneration; degeneration of myocardial vessels; intestinal nemacodiasis; increased splenic hemo poiesis; increased reclculoendothelial proliferacicr in spleen; hepacic lipidosis; diffuse suppurative inflammation adjacent Co .urinary bladder and sex glands. ST0012500 S T 00I250I APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Site 50 Male Rats) Pathology Number 72-108 Days on Test at time of Death____ Gross and Microscopic Observations 456 Gross: Died; advanced autolysis; enlarged, pitted, and mottled surface appearance of kidneys; gastro intestinal contents consisting of dark black material, suggestive of uremia. Microscopic: Pronounced chronic nephropathy with mineralization; secondary parathyroid hyperplasia; myocardial degeneration with mineralization; aortic mineralization. 72-137 485 Gross: Died; slight autolysis; mass in left atrium and ventricle. Microscopic: Proliferating neoplastic mass along endocardial aspect of heart, tentatively diagnosed as cardiac rhabdomyosarcoma; atrial thrombosis; increased splenic hemopoiesis; pulmonary ex foliation of heart failure type cells; metastasis of tumor to thoracic lymph nodes. 72-106 504 Gross: Died; depleted adipose tissue; enlarged, dilated kidneys; aorta dilated; darkening of lungs possibly post-morcea; tesces decreased in size. Microscopic: Pronounced chronic nephropachy with mineralization; myocardial degeneration with mineralization; degeneration of myocardial vessels; cesticular atrophy; aortic minerali zation; lung mineralization. ST0012501 STOOI 2502 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE 3EING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at tine of Death Gross and MicroscoDic Observations 72-109 513 Gross: Killed; overgrown incisors and necrccic . area of hard palace; pale, pitted kidneys with numerous cyscic spaces. Microscopic: Moderate chrcnic nephropathy; degen eration of myocardial vessels; increased splenic hemopoiesis; secondary parathyroid hyperplasia; periarteritis; peribronchiolar lymphoid aggre gates; one focus of alveolar fibrosis and cholesterol cleft formation. 72-113 529 Gross: Died; moderate autolysis; moccled surface of kidneys. Microscopic: Pronounced chronic nephropathy; hepatic congestion: increased splenic hemopoiesis; myo cardial degeneration; vacuolacion of adrenal cortical cells; periartericis, dystrophic miner alized focus in lung. 72-111 536 Cross: Died; subcutaneous nodule weighing 7.4 gn, 2 cm in diameter; lungs appear darkened; en larged kidneys with moccled surface appearance. Microscopic: Pronounced chronic nephropachy; myocardial deneneracion; degeneracion of myo cardial blood vessels; increased splenic hemo poiesis; hepatic congestion; subcucaneous fibro adenoma . ST0012502 STOOI 2503 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY UITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at time of Death Gross and Microscopic Observations 72-112 543 Gross: Died; moderate aucolysis; overgrown incisor; enlarged kidneys, mottled surface appearance; dilated renal pelvis. Microscopic: Pronounced chronic nephropathy with cyst formation; hepatic foci of inflammatory cells; increased splenic hemopoiesis, increased pigment in spleen; degeneration of myocardial vessels; vacuolation of adrenal cortical cells; aortic mineralization. 72-118 547 Gross: Rilled; moribund; corneal opacity; subcutaneous nonfibrous mass adjacent to ear canal. . Microscopic: Zyrnbal gland carcinoma; moderate chronic nephropachy; increased pigmenc in spleen; vacuolation of adrenal corcical cells. 72-120 72-121 556 556 Gross: Died; moderate autolysis; enlarged pituitary; excessive mammary development. Microscopic: Pituitary adenoma; pronounced chronic nephropachy with mineralization; myocardial de generation; degeneration of myocardial vessels; aortic mineralization; isolated foci of in flammatory cells in lungs; subcutaneous galactcce! Gross: Died; kidneys pitted and roughened; inelastic aorca; nodular areas in pancreas. Microscopic: Moderate chronic nephropachy; myocardial degeneration; aorcic mineralization; no change in pancreatic tissue examined. STOO12503 S T 00I2504 APPENDIX A (Continued) CROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at time of Death _____________ Gross and Microscopic Observations 72-123 604 Cross: Killed; large subcucaneous mass 7x7x4 cm weighing 109 grams; kidneys slightly pitted. Microscopic: Subcutaneous fibroma; pronounced chronic nephropathy. ' 72-125 618 Gross: Killed; moribund; enlarged, pitted, mottled surface of kidneys with multiple cysts; enlarged parathyroid glands; multiple reddened and golden brown areas of the lungs; deminerali zation of the bones; Microscopic: No microscopic examination conducted on tissues. 72-127 631 Gross: Died; advanced autolysis; enlarged parathyroids: enlarged, pale, pieced kidneys with mottled surface; hydrochorax; lungs edematous; aorta and aortic arch had loss of elasticity; heart dilated; testes decreased in size and flaccid. Microscopic: Pronounced chronic nephropathy with mineralization; myocardial degeneration with mineralization; degeneration and mineralization of myocardial vessels; thyroid adenoma; peri arteritis; aortic mineralization; exfoliation of heart failure cells uichin alveoli of lung; mineralization of lungs. ST0012504 S T00I2505 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Site 50 Male Rats) Pathology Number Days on Test at time of Death Gross and Microscooic Observations 72-129 634 Cross: Died; left corneal opacity; enlarged, pale, moccled surface of kidneys with cystic structures; decreased adipose tissue; aorta distended with los of elasticity; testes decreased in sice and flacci stomach contained red material; mesenteric lymph nodes tortuous and nodular. Microscopic: Pronounced chronic nephropathy; myoc2rdis. degeneration and inflammation; periarteritis; testicular atrophy; adrenal cortical hyperplastic nodule; vacuolation of adrenal cortical cells; secondary parathyroid hyperplasia; increased reticr endothelial proliferation in spleen; pertbronchiole lymphoid aggregates; edema of lymph nodes; aortic '. mineralization. 72-128 641 Cross: Died; slight autolysis; darkening of lungs; enlarged pituitary (assumed pituitary adenoma). Microscopic: Moderate chronic nephropathy; myocardial degeneration; postmortem congestion of lungs; pituitary unavailable for examination. 72-132 652 Gross: Killed; moribund; bilateral, partial corneal opacity; moccled surface appearance of lungs; enlarged, pitted, mottled kidneys with cystic structures; mesenteric aneurysm; tesces decreased in size and flaccid; anemic appearance of cissues. Microscopic: Pronounced chronic nephropathy; increased splenic hemopoiesis; vacuolacion of adrenal cortica cells; periartericis; myocardial degeneration; aortic mineralization; increased, alveolar macro phages; proceinaceous material and fibrosis of alveolar walls; thoracic Lymph nodes edemacous; testicular atropny. ST0012505 STOOI 2 5 0 6 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at tine of Death ________________ Gross and Microscopic Observations 72-133 665 Gross: Died; moderate autolysis; enlarged prostate; _ mesenteric vessels diiaced; pitted, cystic cortex of kidneys; enlarged parathyroids; gastric auccsa mineralization; testes decreased in size and flacc; Microscopic: Periarteritis with thrombosis; pronounced chronic nephropathy; testicular atrophy, myocardial degeneration; secondary parathyroid hyperplasia; vacuolation of adrenal cortical cells; suppurative prostaticis; increased alveolar macrophages (heart failure cells) In lung. 72-135 697 Gross: Killed; moribund; enlarged pituitary with a darkened area (adenoma); mottled surface of kid neys; two nodular structures in mesentery (4-5 mm) (6-7 mm); a focal reddened area in one adrenal gland. Microscopic: Moderate chronic nephropathy; increased splenic hemopoiesis; increased pigment in spleen; myocardial degeneration; adrenal cortical hyper plastic nodule; adrenal hematocyst; edematous mesenteric lymph nodes; pituitary unavailable for examination. 72-136 . 728 Gross: Died; enlarged, pale, moctled surface of kidneys; enlarged parathyroids; bones soft and friable; large vessels of thoracic and abdominal region dilated with loss of elasticity; atrial thrombosis. Microscopic: Pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; increased splenic hemopoiesis; periarteritis; testicular acrophy; aortic mineralization; atrial thrombosis; increased alveolar macrophages withir. alveoli and focal fibrosis of alveoli. STOO12506 STOOI 2507 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Inicial Group Size 50 Male Rats) Pathology Number 72-143 Days on Test at time of Death Gross and Microscopic Observations 739 Gross: Killed; moribund; emaciated; 25-30 ml serosanguineous fluid in peritoneal cavity; enlarged, pale kidneys with cystic structures; multiple coalescing pale foci in liver; cough focal capsule areas on liver; nodular structures throughout pan creas; cystic structure between liver and stomach, 5 cm in diameter; several focal pale areas in ventricles; aorta distended with loss of elas ticity; enlarged parachyroid glands; thoracic lymph nodes green-brown in color. Microscopic: Pronounced chronic nephropathy with cyst formation; focal hepatic lipoidosis; periarteritis testicular atrophy; aortic mineralization; myo cardial degeneration and inflammation; secondary parathyroid hyperplasia; adrenal pheochronocytoaa; hepatic cyst; pancreatic adenoma. 72-145 756 Gross: Died; advanced autolysis; enlarged, mottled surfaces of kidneys; enlarged parachyroid glands. Microscopic: Pronounced chronic nephropachy; myocardia degeneration; degeneration of myocardial vessels; secondary parathyroid hyperplasia; testicular atrophy; periarteritis; aortic nineralicacion; incra-alveolar aggregates of exfoliated alveolar macrophages (heart failure cells). ST0012507 ST 0 0 1 2 5 0 8 APPENDIX A (ContinuedO GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH ASBESTOS (Initial Croup Size 50 Male Rats) Pathology Number Days on Test at tiae of Death _______________ Gross and Microscopic Observations 72-150 ' 806 Gross: Killed; moribund; emaciated; testes decreased in size and flaccid; decreased adipose tissue; mesenteric lymph nodes darkened; enlarged left adrenal; enlarged, pale, mottled kidneys with cystic structures; mineralization of gastric mucosa; liver pale with subcapsular pale foci; bones demineralized; aorta overdistended with loss of elasticity; enlarged parachyroids; hydro thorax: heart distended and pale; left atrial thrombosis. Microscopic: Pronounced chronic nephropathy with cyst formation; thickening of gastric mucosa with dis tended gastric pics; pulmonary exfoliation of ' alveolar macrophages; focal fibrosis and edema of . lymph nodes; degeneration of myocardial vessels; myocardial degeneration; atrial thrombosis; diffuse keratitis; aortic mineralization; secondar parathyroid hyperplasia; Increased splenic hemo poiesis; vacuolation of adrenal cortical cells; focal pancreatic fibrosis; bilateral adrenal pheochromocytoma; periarteritis; testicular atrophy; decreased secretory content of sex glands pancreatic islet ceil adenoma; increased size of pancreatic islet hepatic cyst formation, diffuse hepatocellular cytoplasmic vacuolation, lipidosis, bile duct hyperplasia, focal necrosis and in flammation and aicrocyst formation. ST0012508 STOOI 2509 APPENDIX a (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STCDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at time of Death ________________Gross and Microscopic Observations 72-104 370 Cross: Killed; corneal opacity; enlarged parathyroid. glands; enlarged, mettled surface of kidneys; enlarged right adrenal (10-15 x normal) , dark and hemorrhagic in color; two nodular structures in pancreas; mesenteric vessels tortuous and sclerotic; testes decreased in size and flaccid; liver had focal pale areas; multiple subpleural circumscribed reddened foci in lungs; loss of elasticity of aorta. Microscopic: Pronounced chronic nephropathy; pituitary adenoma; adrenal pheochromocycoma; pancreatic . islet cell adenoma; pigmentation and edema of lymph nodes; myocardial degeneration and inflam- mation of degeneration of myocardial vessels; increased pigment in spleen; aortic mineralization keratitis; periarteritis; testicular atrophy; decreased secretory concent of accessory sex glands; increased number of reticuloendothelial cells in liver; subpleural aggregates of alveolar macrophages containing hematogenous pigment. ST0012509 S T 0 0 I2510 APPENDIX B GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Time of Death Gross and Microscoolc Observations 72-198 200 Gross: Killed; moribund due to traumatic injury co lower incisor teeth and resultant hemorrhage; toes white areas in lungs, suggestive of aggregates of alveolar macrophages. 72-180 260 Microscopic: Lungs contained aggregates of dust parti cles, sometimes within cytoplasm of alveolar macro phages; adajeent alveolar walls showing minimal hypercellularity; aggregates of dust particles in thoracic lymph nodes; slight chronic nephropachy. Gross: Died; advanced autolysis; increased fluidity of intestinal concents; focal white areas in lungs suggestive of aggregates of macrophages. ' . Microscopic: Pulmonary aggregates of particle-laden macrophages; aortic mineralization; pronounced chronic nephropachy; myocardial degeneration; testicular atrophy; decreased concent of accessory sex glands. 72-154 409 Gross: Killed; Subcutaneous cyst formation within perineal area. Microscopic: Subcutaneous cyst formation and granulom atous reaction; sllghc chronic nephropachy; myo cardial degeneration; isolated foci of hypercellu larity and thickening of alveolar cells adjacent to aggregates of dusc particles, frequently within alveolar macrophages of che lungs; equivocal to vet slighc increase in fibrogenic tissues within thick ened alveoli; pulmonary incerstium and thoracic lymph nodes containing aggregates of dusc particles but not accompanied by any fibrogenic response. ST0012510 ST00I25II APPENDIX B (Continued) CROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEINC USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Tine of Death ______________ Gross and Microscopic Observations 72-152 - 431 Gross: Killed; corneal opacity; overgrown incisors; isolated subpleural circumscribed white foci in the lungs, 1 mm in diaaeter, which disappeared upc inflation with formalin; mesenteric tissue thicker, in one area, 3 cn in diaaeter. Microscopic: pulmonary reaction similar to 72-154; focal aggregates of inflammatory cells in liver; increased splenic hemopoiesis; slighc chronic nephropathy; mesenteric fat necrosis and inflamma tion; pituitary microcyst formation; vacuolation c adrenal cortical cells; diffuse keratitis. 72-169 477 Gross: Killed; moribund; enlarged, mottled kidneys vi:. cystic structures; testes decreased in size; mesenteric vessels dilated and sclerotic; diffuse darkened areas in lungs, one of which appeared fir? and nodular upon palpation; thoracic lymph nodes enlarged and darkened. Microscopic: pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; hepatic lipidosis; periarteritis, testicular atrocr . increased splenic hemopoiesis; secondary parathyroc hyperplasia; adrenal pheochromocycoma; vacuolatior. of adrenal cortical cells; aortic mineralization; pulmonary papillary adenocarcinoma; pulmonary exfo liation of alveolar macrophages (heart failure cell plus edema, hemorrhage and reaction as described fc 72-154. 72-168 505 Gross: Died; autolysis present; traumatically Induced hemorrhage of external nares and nasal turbinates; nasal turbinates hemorrhagic in appearance. Microscopic: increased pigment in spleen; slight chrome nephropathy; myocardial degeneration; degeneration of myocardial vessels; occlusive hemorrhagic rhinitis; sane pulmonary reaction as 72-154. ST0012511 0012512 APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS . (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Time of Death _______________Gross and Microscopic Observations 72-158 507 Gross: Killed; enlarged, pitted kidneys with enlarged pelvic portions. Microscopic: Prounounced chronic nephropathy; pulmonar; reaction similar to 72-154; aortic mineralization; increased pigment in spleen; myocardial degeneratic degeneration of myocardial vessels; secondary parathyroid hyperplasia; vacuolation of adrenal cortical cells; periarteritis. 72-153 529 Gross: Died; moderate autolysis; kidneys roughened on surface. Microscopic: Pronounced chronic nephropathy; secondary parathyroid hyperplasia; cystic retention of material within thyroid; myocardial degeneration; periarteritis, aortic mineralization; pulmonary reaction like 72-154. 72-155 532 Gross: Died; advanced autolysis; fluidity of contents of discal intestinal tract, dark red in color; pale, pitted kidneys, with mottled surface appear ance . Microscopic: Pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; periarcericis, tescicular atrophy; pulmonary reacti: similar to buc less than 72-154. ST0012512 ST00 I 25 I 3 APPENDIX 3 (Concinued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE 3EING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Time of Death______ ____________ Gross and Microscopic Observations 72-157 554 Gross: Died; advanced aucolysis; hydrothorax; lungs filled with fluid. Microscopic: Pronounced chronic nephropathy; myccardia degeneration; degeneration of myocardial vessels; periarteritis; vacuolacion of cortical cells; foes hepatic fibrosis; epidermoid carcinoma of lung with metastasis to thoracic lymph node; also pul monary reaction like 72-164 plus exfoliation of macrophages (heart failure cells). 72-160 531 Gross: Died; moderate aucolysis; one subpleural cir cumscribed pale foci in lung; enlarged, pale kidne' Microscopic: Pronounced chronic nephropathy; myocardial degeneration; vacuolacion of cortical cells; pulmonary reaccion similar to 72-154. 72-162 603 Gross: Rilled; moribund; darkened areas in lungs firm on palpation; choracic lympn nodes containing red pigment; kidneys pitted with mottled surface appea: ance and cyscic areas; enlarged parathyroids; aortic arch dilated and inelastic. Microscopic: Pulmonary reaction similar to 72-154 plus focal alveolar hyperplasia and fibrosis; pronounced chronic nephropathy; increased splenic hemopoiesis; secondary parathyroid hyperplasia; periarcericls; testicular atrophy; aortic mineralization; vacuo lacion of cortical cells. ST0012513 ST00 I 2514 APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE 3EING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number___ Days on Test at Tine of Death ______________ Gross and Microscopic Observations 72-163 ' 625 Cross: Died; hydrochorax; several subpleural circumscribed pale foci up to 4 on in lungs; kidneys roughened over surface. Microscopic: Pulmonary epidermoid carcinoma; also reaction similar to but more pronounced than 72-154; aortic mineralization, myocardial de generation; degeneration of myocardial vessels; secondary parathyroid hyperplasia; pronounced chronic nephropachy; periarteritis. 72-165 630 Cross: Died; enlarged, pitted kidneys; enlarged sub- lumbar lymph nodes. . Microscopic: Pulmonary alveolar adenomatous pro .. liferation; also pulmonary reaction described for 72-154; pronounced chronic nephropathy; myo cardial degeneration; vacuolation of cortical , cells; edema and pigment within lymph nodes; secondary parathyroid hyperplasia. 72-167 652 Gross: Killed; two subcutaneous masses weighing 64.7 grams and 36.3 grams; pitted kidneys with mottled surface appearance. Microscopic: Subcutaneous neurofibroma; hyperplasia of adrenal medullary cells; pronounced chronic nephropachy; focal inflammation in portal triads and lobules of liver; pulmonary reaction similar to 72-154 plus more lung fibrosis and focus of alveolar adenomatous proliferation. ST0012514 ST00I25I5 APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Tine of Death Gross and Microscopic Observations 72-170 652 Gross: Died; emaciated; enlarged parathyroids; en larged parathyroids; enlarged, pale kidneys with mottled surface and cystic spaces; gastrointestir.c tract darkened; gastric wall Chickened; aorta dilated and inelastic; no adipose tissue; testes decreased in size. .72-171 658 Microscopic: Lung reaction like 72-154 plus focal fibrosis and alveolar hyperplasia; aortic ainerali ration; secondary parathyroid hyperplasia; myo cardial degeneration and fibrosis; pronounced chronic nephropathy; periarteritis. Gross: Died; enlarged kidneys with cysts and mottled' surface; enlarged parathyroid; enlarged pituitary dark in appearance; turbinaces congested or hemorrhagic. , Microscopic: Focal pulmonary lung fibrosis and alveola ' hyperplasia; also reaction like 72-154; pronounced chronic nephropachy; hepatic portal inflamination; myocardial degeneration; pituicary adenoma; parathyroid not examined. ST0012515 ST00 I 2516 APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test ac Time of Death Gross and Microscooic Observations 72-172 661 Gross: Killed; two subpleural circumscribed gray-white foci in lungs 3 cm in diameter; nodular structure in duodenal wall; two nodules in mesentery 1/2 cm and 2/3 cm in diameter; thoracic lymph nodes enlarged; pale kidneys. Microscopic: Pulmonary reaction like 72-154 plus focal alveolar fibrosis and hyperplasia; aortic mineralization; edema of lymph nodes; aoderace chronic nephropathy; myocardial degeneration; increased splenic hemopoiesis; periarteritis; secondary parathyroid hyperplasia; adrenal pheochromocytoma; mucinous adenocarcinoca of small . intestine; thyroid adenoma. 72-175 675 . Gross: Died; moderate autolysis; enlarged, pale, mot tled kidneys; enlarged parathyroids; mesenteric blood vessels dilated; left atrial thrombosis; pulmonary consolidation; tesces decreased in size. Microscopic: Pronounced chronic nephropathy; nyocardia degeneration; vacuolation of cortical cells; adrenal hematocyst; periartericis; left atrial thrombosis; secondary parathyroid hyperplasia; aortic mineralization; pulmonary reaction like 72-164 plus exfoliacion of alveolar macrophages. ST0012516 LIQZIOOIS APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Time of Death _______________Gross and Microscopic Observations 72-179 . 697 Gross: Killed; moribund; multi-loculated hemorrhagic mass within mesentery, 10 cm in diameter; firm nodular structure in pancreatic area of mesentery; enlarged pale spleen; liver pale; aorta with loss of elasticity. Microscopic: Incraabdoninal malignant schwannoma; aort mineralization; pulmonary reaction similar to but less chan 72-164; increased splenic hemopoiesi slight chronic nephropachy; myocardial degeneracic: focal aggregates of reticuloendothelial cells in liver; pancreatic lslec cell adenoma. 72-190 ' 697 Gross: Died; subcutaneous multi-nodular structure ' weighing 21 grams; hydrochorax; heart distended and containing blood clots; focal darkened area ' In lung, 5-7 mm in diameter; enlarged kidneys; enlarged left adrenal; aorta dilated with loss of elasticity; mesenteric vessels tortuous and sclerotic. Microscopic: Lung reaction Includes that described for 72-154 plus alveolar hyperplasia, squamous metaplasia and fibrosis; aorcic mineralization; pronounced chronic nephropathy; periarceritis; myocardial degeneration; degeneration of myo cardial vessels; focal hepatic portal fibrosis; subcutaneous fibroma; adrenal pheochroraocycoma; secondary parathyroid hyperplasia; edema and congestion of lymph nodes. ST0012517 lST00 I 2 5 8 APPENDIX S (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYINC WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Time of Death______ Gross and Microscopic Observations 72-181 721 Gross: Died; tnoderace aucolysis; absence of adipose tissue; fluidity of incestinal tract contenc; mineralization of gastric mucosa; enlarged, pale" kidneys with mottled surface appearance; one cyst 3 mm in diamecer; liver pale; enlarged parachyroic nodular structure in pancreas 1/2 cm in diameter. . 72-183 . 721 Microscopic: Lung reaction like 72-154 plus focal area of alveolar epichelial adenomatous proliferation; aortic mineralization; pronounced chronic nephropa' focal hepatocellular vacuolation and lipidosis; focal portal fibrosis in liver; myocardial degener. tion; degeneration of myocardial vessels; peri arteritis; testicular acrophy; vacuolation of ' cortical cells; secondary parathyroid hyperplasia; pancreatic adenoma. Gross: Died; emaciated; advanced aucolysis; fluidity o: incestinal tract; mineralization of gastric mucosa: enlarged, pale, mottled kidneys with cystic areas. Microscopic: Pronounced chronic nepnropachy; periarcer: tis; testicular acrophy; myocardial degeneration; aggregates of reticuloendothelial cells in liver; increased splenic hemopoiesis; secondary parachyrot hyperplasia; adrenal pheochromocytona; aortic min eralization; pulmonary reaction similar to 72-154. ST0012518 ST00125 I 9 APPENDIX 3 (Continued) CROSS AND MICROSCOPIC. OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Site 50 Male Rats) Pathology Number Days on Test at Time of Death _______________Gross and Microscopic Observations 72-185 . 724 Gross: Killed; moribund; scattered subpleural circumscribed pale foci in lungs up to 2 mn in diameter; aorta dilated with los3 of elasticity; enlarged parathyroids; liver pale with multiple foci up to 2 mm in diameter on all lobes; uremic gastritis; enlarged, pitted, pale kidneys with multiple cystic structures; decreased adipose tissue; testes de- creased in size and flaccid. MMicroscopic: Pulmonary reaction like 72-154 plus more exfoliated macrophages; pronounced chronic neph ropathy; focal reticuloendothelial cell aggregates and lipidosis in liver, increased splenic hemo poiesis; myocardial degeneration; degeneration of myocardial vessels; pancreatic islet ceil adenoma. 72-195 732 Gross: Killed; moribund; circumscribed area 2 cm in diameter having the consistency of an epidermal inclusion cyst; mesenteric vessels containing mul- tiple nodular areas; fluid in abdominal cavity; enlarged, pale kidneys with nulciple cystic areas; righc kidney containing a pale circumscribed mass 1 cm above the surface and 1 1/2-2 cm in diameter; fluidity of contents of intestinal tract and stomach; subpleural pale areas in the liver; en larged, pale parathyroids; hydrochorax; focal red dened area in pituitary; left atrial thrombosis; thoracic lymph nodes enlarged and red-brown In color; two subpleural circumscribed pale foci 1/2 cin diameter. Microscopic: Pulmonary reaction similar to 72-154, plus increased exfoliated alveolar macrophages and focal alveolar fibrosis; aortic mineralization; adrenal hematocv3t and vacuolacion of cortical cells; pertarteritis and thrombosis; pronounced chronic ne phropathy with cyst formations and fetalization of epithelium lining tubules; testtcular atrophy; hepatic lipidosis and extramedullary hemopoiesis; granulomatous colitis; secondary parathyroid hyper plasia; myocardial degeneration; atrial chrcmbosis; pituitary adenoma. ST0012519 ST00I2520 APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Site 50 Male Rats) Pathology Number Days on Test at Time of Death______ Gross and Microscopic Observations 72-200 - 732 Gross: Killed; moribund; enlarged, hemorrhagic pituit. enlarged, pale prostate, with purulent material; enlarged, pale kidneys; enlarged parathyroids. Microscopic: Picuitary adenoma; aortic mineralization: ' vacuolation of adrenal cortical cells; periarteri: secondary parathyroid hyperplasia; myocardial de generation and fibrosis; moderate chronic nepnropc with mineralization; focal biliary hyperplasia in liver; increased splenic hemopoiesis; pulmonary reaction like 72-154; prostace not examined. 72-192 812 Gross: Died; enlarged, dark kidneys with scattered pale foci; enlarged liver; 2 subpleural circus- ' scribed gray foci in lungs; pinpoint in size. " Microscopic: Pituitary adenoma; similar pulmonary reaction to 72-154; focal pancreatic fibrosis; pronounced chronic nephropathy; liver congested. 72-193 835 Cross: Died; subcutaneous mass 2.5 cm in diameter; hydrothorax; left atrial thrombosis; enlarged kid neys mottled in appearance; one thyroid enlarged and dark; nodular area projecting off pituitary; aorta with loss of elasticity. Microscopic: Myocardial degeneration; degeneration of myocardial vessels; atrial thrombosis; pronounced chronic nephropathy; hepatic cellular vacuolation and cyst formation; thyroid adenoma; pulmonary reaction similar to 72-154 plus focal alveolar fibrosis; subcutaneous carcinoma; pituitary not examined (adenoma?); diffuse keratitis. ST0012520 ST00I252I APPENDIX B (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Tine of Death______ Gross and Microscopic Observations 72-196 843 Gross: Died; moderate autolysis; testes decreased in size and flaccid; red mass in one testis 5 mm in diameter; suppurative prostatitis; decrease of ' adipose tissue; enlarged, dark kidneys, nottled in appearance; subcapsular hepatic pale pinpoint foci, 1 mm in diameter; hydrothorax; left atrial thrombosis; enlarged thoracic lymph nodes. 72-197 865 Microscopic: Pulmonary reaction like 72-154, plus more alveolar macrophage exfoliacion and focal fibrosis of alveoli; aortic mineralization; focal amyloidos and reticuloendothelial cell proliferation in liver; pronounced chronic nephropachy; myocardial degeneration; atrial thrombosis; suppurative pros tatitis and urocysticis; vacuolacion of cortical cells and adrenal hematocyst; thyroid adenoma; ' testicular atrophy; interstitial.cell tumor of testicle. Gross: Died; corneal opacity; nodular area in left lung; firm on palpation; firm nodular structure on primary bronchus, 8 nm in diameter; liver motel in appearance with scattered subcapsular pale foci up to I mm in diameter; pale, pitted kidneys. Microscopic: Pulmonary papillary adenocarcinoma of lung; focal alveolar adenomatous proliferation and alveolar hyperplasia; some pulmonary reaction like 72-154 plus more alveolar fibrosis; pronounce: chronic nephropachy; thyroid adenoma; secondary parathyroid hyperplasia; vacuolation of adrenal cortical cells; splenic congestion; myocardial de generation; hepatic congestion and bile duct hyperplasia; aortic mineralization. 72-199 - No animal available for examination,.probably due to cannibalism or loss of rac. ST0012521 CROSS AND HISTOPATHOLOGIC OBSERVATIONS ON NOM-PU1.MONARY T IS S U E S OF MALE RATS K IL L E D AT 7 , 8 OR 13 MONTH T IM E INTERVALS DURING IN H A LA T IO N STUDY W ITH ASBESTOS Data lis te d as n u m b e r o f ra ts in each group show ing th e v a rio u s o b s e rv a tio n s . ux luaaSfd jo uof^BxnmnDDB xB30i o o o o c SB3J3UEd jo sTSojq-r; -[B-p'psaaauT xB30I O O OO rH O 31}2txs * sfsaxodoaiaq ojusxds pssBsaoux O o o o n n uoT3Banrox jut pun uoj^Baauaaap xBTP-tB30^m TBOOj --i --< o CM -- S33.C001Bd3q 31BJ3U323P O} 3U33Bppe SXX33 jCjo^BnmrBXjuj jo s31b93t32b p33BX0SI OO rM o HN SJSBTpo^Bia3U XbuTTS33UX -h O O O <T 33Bi3potn 'XqaBdouqdsu OTuauq;) O O O O -i o jq3xxs `Xqiedoiqdsu ojuoaq^ O O O O -y m sjAXsd xBU3i UT sajsodap pBzjx^311?;* -HO -HO O C xsajnqni BU31 UT sajsodsp pazTxenaujp; CM rH O O O O jCsupt'y uj s33b93j93b XT33 ^-lorieinaex jui cm O O O u" SUOJ1B3JOJ jsbo jEXnqru XBU3H lT\ XX^ax33 PUB spAjad x503-1 J uoiis^BXTQ --I O O C O O sAsupjq jo aoejjns paxdJOH O O CM rH rH rH q333] JOSTOUJ UAlOi3^3A0 O O O O rH 3Si<D03CUI3L( XBU3-1PV O O O O rH si try jo jaqanq SJ~\ tn lH cO dnojQ 1U3U13E3JX <rt --H O o 1H u CO w 17 C -3 o CO u< in ^H 0 o 4J u .0 *J 1) c o (/I u< CO O o 4H H CO 3 c o v> u< XEATJ3UT 3U1JX r-* r-* n ro ST0012522 CO CSJ LO CVJ o o I-- CO -J 10 (--A >a- ^ !r I<>eUe-J <>tu--ti <foF= Co --Xo t<-- X<o ou__>J: tCoO M*l riqiajn pur jappriq Xjrui jn oa ooaprfpa rt~333rs pa:r; 30*3 a;; i pu.'j ja*77 ci taaXpcjirdaq pairionpeA ;c snuo* tooupf jCjciI r>3 1 i xasjo? x9jpr ;o ?|npou d i 3 sr i d J adXjj T;odott*q pxuajdc pasca33uj ua*ids ux 3uacS;d snouaaosroa^ paBea3DUi rx^npac xruaipe 30 a^npou 073sridjad^H ruouapc p-joiXi^x B0 3XDomoac| ooayd jcuajpy risctdjadXM p703Xi23C3*d /jcpu03s TTd 1WP13303 iuajpr 30 uoijrxonoa^ itqdoisr jxn3|3Bl )DS1S*A 373*20133 pur 9 3 3 0? 0 0 I 3 2 T T f 3 U T K tX37a3 3TlJd 3*tr\P7 3*J 37 J3U*SK papunoaoid ` Xq3?dojL|du 071103173 *3tjapoa ` X43?doii|dau 37003173 uo73*mTXju7 pu* uoxirjauaBap 373*d'q 7*004 *7saca8o3rejadB pc*a33Q *7*o3<;73 pur uotarooiri;ui *uoj3jaua8p x?ipj?poX^ ?7*ba poojq 7?7p3poXc ;o uoxaejauaBafl japprxq XJaujjn 03 3ua3*fpr str^ Bsaosqr anoauc^nonq flAfad jvuaj pa3ri7Q O O 3070^ ^0013 3 j7 7 uo73PCJ3U03 X3e37n37d uo aaa* pauappaj 7*304 cpurjg xc pur ta3t3 70 azys pacea33aQ 77m 0133**8 30 uo;3r;77*3au7X *U73jCqdJocl x**u *0073*70*^3 5p70aXt|3ri*d jo aauauiooad paseaaouj 3a77 uT T30J 3?xnsdToqns axed axdxsxnj; -- pu?x* xruaap* 3 *rT* T asraioui ~ Xaupx^ u; 3CX3 Xaup7> pauaqSnoa pur pa8jxu *Trd X370cdo 1**02031^ pau7wax3 ase-g 30 jaqonj; dnoj^ ouacieaa^,;^ si;3uo^ u; i*.\j3u; =7i; D n ta .lifte d *8 number o f r n t* In each group showing the v a rio u s o b s e rv a tio n *. ST0012523 STOOI 2524 APPENDIX e INCIDENCE Of TUMORS IN GROUPS OF 50 MALE RATS USED AS CONTROLS IN INHALATION STUDY WITH ASBESTOS Morphologic Type of Tumor and Probable Origin Neural craniopharyngioma Subcutaneous fibroma Subcutaneous fibroma Pituitary adenoma Intraabdominal sarcoma Pancreatic adenoma Cardiac rhabdomyosarcoma Subcutaneous fibroadenoma Zymbal gland carcinoma Pituitary adenoma Subcutaneous fibroma .. Thyroid adenoma Pituitary adenoma . Pituitary adenoma Adrenal pheochromocytoraa Pancreatic Adenoma Adrenal pheochromocytona Thyroid adenoma Pituitary adenoma Adrenal pheochromocycoma Pancreatic islet cell adenoma Pituitary adenoma Adrenal pheochromocytoma Days on Test at Time of Death of Rat With Tumor 109 136 409 409 413 420 435 526 547 556 604 631 641 697 739 739 742 742 742 306 306 370 870 Rat Number 72-146 72-134 72-115 72-131 72-L48 72-149 72-137 72-111 72-118 72-120 72-123 72-127 72-123 72-135 72-143 72-143 72-142 72-142 72-138 72-150 72-150 72-104 72-104 Includes all tumors diagnosed in racs dying durin g study or at the interim kills. ST0012524 STOOI 2525 APPENDIX P INCIDENCE OF TUMORS IN CROUP OF 49 MALE RATS USED IN INHALATION STUDY WITH ASBESTOS Morphologic Type of Tumor and Probable Origin Days on Tesc at Time of Death of Rat With Tumor Pulmonary papillary adenocarcinoma Adrenal pheochronocytoma Pulmonary epidermoid carcinoma Pulmonary epidermoid carcinoma Subcutaneous neurofibroma Pituitary adenoma Adrenal pheochronocytoma Mucinous adenocarcinoma of small intestine Thyroid adenoca Intraabdotninal malignant schwannoma Pancreatic islet cell adenoma Subcutaneous fibroma Adrenal pheochromocytoma Pancreatic adenoma Adrenal pheochromocytoma Pancreatic islet cell adenoma Pituitary adenoma Pituitary adenoma Pulmonary mucinous and adenomatous carcinoma Pituitary adenoma Adrenal pheochromocytoma Thyroid adenoma Thyroid adenoma 477 477 554 625 652 658 661 661 661 697 697 697 697 721 721 724 732 732 742 742 742 742 742 Rat Number 72-169 72-169 72-157 72-163 72-167 72-171 72-172 72-172 72-172 72-179 72-179 72-190 72-190 72-181 72-183 72-185 72-195 72-200 72-136 72-136 72-136 72-186 . 72-138 ST0012525 STOOI 2526 APPENDIX F (Continued) INCIDENCE OF TUMORS IN GROUP OF 49 MALE RATS USED IN INHALATION STUDY WITH ASBESTOS Morphologic Type of Tumor and Probable Origin Days on Test at Time of Death of Rat With Tumor Undifferenciaced sarcoma attached to urinary bladder Adrenal pheochromocycoma Pituitary adenoma Pituitary adenoma Thyroid adenoma Subcutaneous carcinoma Pituitary adenoma Thyroid adenoma Testicular interstitial cell cunor Pulmonary papillary adenocarcinoma Thyroid adenoma ' 742 742 742 812 835 835 835 843 843 865 865 Rat Number 72-188 72-189 72-189 72-192 72-193 72-193 72-193 72-196 72-196 72-197 72-197 STOO12526 <01 OS Ui H Vi "X. < Z >-- c/i z 00 oC r-A H to < 3J > CO QS 00 W< t/1 CO li. oc o> r-A Z oz o c-- _J 00 o zz pC < 1-- O- w o< H- z t/1 < >--i *T zz r-< Q os o u. 00 CO 00 oZ OS o o OS H PS o"* azCJ oo < Q UonJ <z f-- < o >- *- -3 i0S O t; 01 -3 1 <3J o I 1 01 H 0 o K*-l > 4-4 o r4 9 O --1 >9 W TJ 01 0 - S C ID H 0 >> C9 ^4 u u C3 CL o (9 kJ U 01 3 --4 TJ o p--< u m4 4*4 >4H 0 a 3 m4 9 -3 - >> .* TJ TJ 01 9 U 44 O 9 cn a 0) *+ c --4 a. 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CJ X CO C u Cl O T3 --4 C CO c 3oo Z 4- 44 H 4-1 u C- 3 o `J SI 2 c 44 0 W4 3 O OZ c au - 4J <3 cZ 9 o ac C- 3 3 co C U O3 9 C rH 93 4-1 C c CJ o in z "d n3 -ri z 3 EC -4 to C 3 z Si Si 9 10 9 CO CO CO </> oZ >s >, >. h* CO (0 3 9 4-1 a Z c o cc 0 co z z 3 O' nT vT rs to B -J wO H-N >s 9H a 1_1 C3| LJ c 9 e to 4H z TJ C 9 d 4^ o rH >> a <a 4^ CO >4 CJ >s U 9 44 rH CO 44 44 Z co 0 -3 CO 4H 44 O LJ rH si C9 O d ^ O9 c 4-1 3 C ^4 rn n 3 u -o 3C Z9 ou 4-4 44 Z 93 3 rH -a z tn TJ 9 ~ 4-1 U 93 > U-- 9 rH a 9C >*o to C Hi 3 -H z V) -4 O Li z C3 X "3 O - Li to 10 9 44 3 w U) si CO 0 39 d 30 ^H 44 3 O o UZ rH to u X to a 3 *d 9c U9 TJ 9 C ^ CO 9 3 Si rH --4 C rH 71 c 30 44 U Z Si 9 U -O a3 e rH T3 9 O9 p-H 9 N <4h > c 44 9 9 Li - rH CJ Cl 9 C 4-1 U -4 7) O S X CJ . g X 9 7) w S9 3 O ^H 4 cz rH 3 Q 44 -3 si z 9 5k U u r-4 44 3 C 4-1 U H 9 Z -i to 9 L* CJ Z c -- uu . - -H CO rH 4-| 4H 9 U SJ 3 3 44 3 Z Cl uu to 9 d z c Si 3 si 3 C rH 9 3 to c - -4 CO H Z to 3 Si C CO to 9d 9 c 9HZ o TJ o z 44 4* O SJ (0 >> L4 9 3 a9 a O 99 u 9 sO C CJ S4 9 -- ?s Si Si c 3 si <s- O H CO 9 z zo tn to a U to in >s >N 39 Oz r>. un in vO * c > r-* r* m -- -r CO 4/1 c kH un w~> -- CM Psl PM PM CM PM 1 r pm r'l CM rs< PM PM ft- P4, P4. rs P- GO --H CD CD ^--4 ro cn ro co STOO12528 MORTALITY DATA AND MAJOR CROSS AND HISTO PATHO LO G IC OBSERVATIONS IN HAMSTERS USED AS CONTROLS FOR IN IT IA T IO N STUDY OF ASBESTOS t >s -3 u u a cl d 0 G- ^4 cn T3 H X l- B u (1 *3 a3UC Oa ua 3 u 6 UJ kJ <a C1) h *_ o 7) a. u to VI 3 O -a 4J 3 kJ Cl i_> tM U V . T5 H >--4 0) u 4J *o 3 4) 4 to U kJ c 3 d ai ** 3 3 CO c U i-- U >N o U) a. U 3 u k4 to a W Cl u d U> *T3 C m a 'J) . * 4J tn CO a > C o a 3 "3 3 to X to to O a. O o d u co a. o . o3 rs JZ C. *r4j tc *--o okJ aj VI u kt c c <1 d W <--i rs 3 T3 3 Q. o >x s U U JZ JZ zo o V> V kJ <-4 CL 3 . c. >v > >> c. a 6 o B -3 3 tM kJ o c o "3 10 "3 3 -- 3 acO c ou o to " 33 "3 >. kJ JZ z 0 j O 1 kt 3 a3cNd cc u V) a g a c d CO o kj 3 D --4 kJ c. u U ** to <1 (J JD 3 C rHd u d u M Q. o o . U .. c - 05 3 >N 0 d | a ** c. u 2 xj >s ^ -j 1O -> w CJ -3 -- ^ ^H U - O uc wa > co> a o a* o 6 ^ u - o cJ 1 T3 -h 3 -- rj C V- *- <0 - 3 3 to kj to * M C 'J C XJ <S) Q <J tc o U) u CO Z a. >% a o to U C 0 "3 <J X ^ k<J1 to "j 3a H co o 3 v>N 3H D a r-\ fN r-* C >\ o c| rv* a''.iI rsi r> STOOI2529 STOO12529 < -v o0 CO cn c u3 c 0 F4 y SM W4 44 "3 4 cn cn 40 44 V) Q 40 33 "3 to 0 to 3 4 0 U ^4 40 44 3 3a y C o*. 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