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fltioum TOR API'KOVAL tg release; PROPRIETARY TECHNICAL INFORMATION QW e- /733Q 1 P.7l-/o-i3LOl_r ' . Effects From Chronic Inhalation of Asbestos Pipe- r t Mi \ AN A r/tiRE VIA T t' l_j 'Ml OR AbSTRACT Covering Dust in Rats and Hamsters' MIS $ nw aORK FRrv ;OUSLT JNREPORTED B.K.J. Leong, R.J. Kociba, L.E. Frauson, R.W. Lisowe, : 71 rTAL rACE'- !NNCCIUDING L.W. Rampy J -Li--__- 7> nli ; AN-pnrGicUuRfE_S_____ ' 1_'n ' '' This report is to be given to former employee B.K.J. Leong to get ir. shape for publication. 05 u ' When revised, to Amer. Ind. Hyg. Journal RELATED PaTFnTS OB DISCLOSURE F1.a"5D P^OP"ri;- OP ' OCE ;S ES RFLATfcD PEP 05 . CD s' k: KJ 1 j ! sur.cfirs^NS h* a >' * i . At.f't D row ross-p. z prv!i**ERs or this manuscript r `OK 1 fCMN'CAL ASPECTS / :! 'mpoc*' T''Verkema -<>" - 0 'CAi.Tf. . Miv,BOi.VLNUt RES. name V. K. Rowe Toxicology Research Q Tf-.HNCLCGY 'ENTER ,, L'ff, _______ _________ 0 C : rt E R ^ -P' * 8 T'l Jd'OOl :1 CUTES' iN4 COnO* A(^Nr. IH.S vanUSCRiPT ARE TO 3 DIRECTED TO L. W. Rampy 1803 W Midland 2j C - It ** * >$ C C' >>nM .. 55CoCc. .r f a? mi . : ' r UJ* P' - cr:. f't VG-" ;N0! ^ j ; , Toxicology Research 10/11/76 \r-- c PMONL:: 636-626.0.. DATE M '.CEr . 10/21/.7-_ iu'ni and s* -cpe; of the DF^A^TmENTal APPROVAL FOR RClFaST I J . - r- ~ J ` ...J . J . . . : Of ' ^ DEPARTMENTAL SFLCCTtON OF RVtJ>C.F^ .iU riA Tr . ;^a t ETT--^-- - . f: i : J. R. Peele, 2020 it l' > ' N {' ( J. s. Laing, 2020 r:. AT j_- ' J_ V. K. Rowe, 1803 :i*i#j i . - to C t N' kA , */' EEC: 1 cm .**-> !*M' \`An l DSN * ,, r * a:. (O > hYp,,i will a v#r b> him Se*id *h*s si^tifcu in midland cw FEB 1 i 1967 'CCOmmENDAt jQNS 0 V.USI [_] Rf i i Air Ar:i:: . .,.nges 0 do t:o7 pe l r ai.r b * r. *; a . w r* U-, I cr*. - ' lr.r . 4 Rrr,0R; i.:u n; .'IN' r (t mst 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 4S640 May 11, 1976 407123 $ T 0 0 0 IM i9 ABSTRACT 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. A smaller group of hamsters exposed under these conditions experienced an earlier onset of mortality than control hamsters not subject-id 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. 407124 : STQ00I450 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. 407125 l Sh I 0001S ! 3 Most of the asbestos inhalation experiments to date were conducted using relatively pure minerological types of dust. The findings in rats 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 thu life of the animals. 407126 : STOOD I 452 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 submicron 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% MgCOH^ 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 Pyrex 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 407127 tS T 000 I 453 STOOOl'tS 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 pm 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. 407128 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 407129 STOOD 1455 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 Interim kills of 5 control rats and 5 rats exposed to as bestos were conducted at the end of the 7-month exposure, 407130 1ST000 11458 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 T000 I 457 co 407131 8SM 0001S 9 (Masson's Trichrome stain) and of rcticulin (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 the lungs and thoracic lymph nodes to define the response to repeated inhalacion 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. Herutovylin and eosin stained sections of these tissues were routinely prepared f3r 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. 407132 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. 407133 ST00 0 I 459 j ST000I 460 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 camples of asbestos dust indicated that the average dust concentration in the chamber atmosphere was 85 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 um. However, direct measurement of the dust particles suspended in air with a ligh-u 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 Durin ich exposure period, signs of slight ocular and nasal `citation were observed in both rats and hamsters. The animals buried their muzzles in the hair coat. The animals remained inactive until they were removed from the 407134 i ST00 0 I li 6 I 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 cf 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. 407135 STOOD I ti62 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 mm in diameter. Microscopically, the principal changes were isolated aggre gates of dust-lade macrophages within alveoli. Many of the affected alveoli w^re 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 fibrogenic 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 on non-pulmonarv tissues examined at the interim examinations. 407136 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 evalyi'tion 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-pulmonary, tissues revealed no alterations considered related to exposure to the asbestos dust. * A total of 15 rats per group had been killed for interim examinations at 7, 8 and 13 months of the study and one ex posed rat was missing. **Appendixes E and F contain a listing of the temporal in cidence of all tumors in control and asbestos exposed rats respectively. 407137 ST000 I 463 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 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 asbestos-exposted hamsters respectively. 407138 W IOOOIS ST000I 465 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 407139 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. 407140 9 9 *110 001 :s 18 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 pulmonar-' 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 a_l. 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 lung to a similar concentration. The latent period for tumor development was practically the same (16 months) in both studies. In si. m ilar studies by Reeves et al. 22 (1971). rats were ex posed to chrysotile dust at a lower concentration of 48 mg/M? 407141 I ST000IU67 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 al^ noted a low incidence of pul monary squamous-cell carcinomas. Two of the factors believed to be associ '.ted 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 y particles and very few fibers (Fig. 2). Thus, the production of pulmonary carcinoma in the present experi ment supports the conclusion of Kogan et al., who considered fiber length or fibrous structure as non-essential for pathogem. c.ity. 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 chrysotile samples of Gross et^ al., 21 and Reeves et al. 22 The high content o.i 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 22 which contained rather low concentrations of nickel and cobalt indicates that the 407142 ST000I468 20 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. CD CD cn UD 407143 REFERENCES Hendry, N. W. The Geology, Occurrences, and Major Uses of Asbestos. Ann. New York Acad. Sci. 132:12-22, 1965. Cooke, W. E. 1025, 1927. Pulmonary Asbestosis. Br. Med. J. 2:1024- 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 79-81. 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-Abdominal Neoplasia. Ann. New York Acad. Sci. 132:519-525, 1965. ' ST000I4*70 407144 IZ.'llOOOiS 7. Selikoff, I. J., et al. Mortality Experiences of Asbestos 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, 1. J., Hammond, E. C., Churg, J. Asbestos Exposure, Smoking and Neoplasia. JAMA 204:106-112, 1968. 10. Cooper, W. C. Asbestos as a Hazard to Health. Arch Environ. Health 15:285-290, 1967. 11. Selikoff, I. J., Churg, J., Hammond, E. C. Relationship Between Exposure to Asbestos and Mesothelioma. N. Engl. 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. Mesothelioma^ in Rats Following Inoculation with Asbestos. Brit. J. Cancer 23:567-581, 1969. 407145 15. Berry, G. and Wagner, J. C. Application of a Mathematical Model Describing the Tine of Occurrence of Mesotheliomas in Rats Following Inoculation with Asbestos. Brit. J. Cancer 23:582-586, 1969. 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. 18. Cralley, L. J. and Lainhart, W. S. Are Trace Metals Associated with Asbestos Fibers Responsible for the Biologic Effects Attributed to Asbestos? J. Occnp. Med. 15(3):262-266, 1973. 19. Dixon, J. R. et al. The Role of Tracei 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):11751187, 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. 407146 ZL'i I0G01S r 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 Fihrogenic 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, 1968. L'y CD CD CD CO 1 407147 TABLE 1 ATOMIC EMISSION SPECTROGRAPHIC ANALYSIS OF ELEMENTS PRESENT IN THE BALL MILLED ASBESTOS SAMPLE USED IN INHALATION EXPOSURE OF RATS AND HAMSTERS Element Weight I of Sample o 00 A1 Ba 0.008 B <0.05 Ca 0.4 Cd <0.05 Co 0.004 Cu <0.001 Cr 0.07 Fe 2.0 Mg 11.0 Mn 0.04 Ni 0.09 Pb 0.01 Si 10.0 Sn <0.01 Sr <0.001 Na 1.0 K <2.0 Ti 0.5 V <0.005 Zn <0 " <1L'i I 0001S 407148 GROSS PATHOLOGIC OBSERVATIONS ON PULMONARY TISSUES OF MALE RATS KILLED AT SERIAL TIME INTERVALS DURING INHALATION STUDY WITH ASBESTOS in soqsaqsv CN m -[caquoo CN o oo CN ooo o m soqssqsv in m [caquoo in rP h o o o OO O O co soqsaqsv in co Xojquoo in H O O O oo o o soqsaqsv in [OJ^UOD in mO o o IT o I XI A 3a to to E en CO cn c oH i--Q>1 cfl Cr c rH e3 3 rH o Pto PA4-> (0 > a -H to E 4-1 c H u to Q) g 0 p c o WCD XI P CN 0) to o 10 A s XoI P 1 o 1 to P V to p <0 c H PiH 3 10 p 0 p 10 3 -P co 'O A U O 01 O cc 0 P 4P a QJ 3 Pp0) P EH u t--t cO 13 rO to Cp o 3 0) fp P TJ (0 IQ P rP PC CO P O .lp: p E 3 o p p a P o P c 3 CO o co E' Xu s aaa to c -oHU <0 > p 0) in XI o cn O3 PH to > 0) iz p WCD .C5O to a 3 0 p O' au <0 W e H to P f0 P U-l 0 P 0) X 3 C co CO a 0J p to p rp (0 p to a 407149 ST00 0 I f| 75 .TABLE 3 HISTOPATHOLOGIC OBSERVATIONS ON PULMONARY TISSUES OP MALE RATS KILLED AT SERIAL TIME INTERVALS DURING INHALATION STUDY WITH ASBESTOS Time IntervalIn Months 7 Treatment Group________ .__________________ Number of Rats___________________________ Microscopic Pulmonary Observations; H 0 u u c 0 u 5 Alveoli and Alveclar Membrane Isolated focal aggregates of particle- laden alveolar macrophages 0 Isolated focus of inflammatory cells in lungs 0 Isolated aggregates of alveolar macrophages 0 Isolated focus of alveolar epithelial hyperplasia 0 Isolated focus of hypercellularity of alveolar wall 0 Isolated foci of hypercellularity and thickening of alveolar walls adjacent to particle-laden alveolar macrophages - very alight 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 andmucinousareas 0 Sarcomatous-like proliferation of tissue in lung and thorax 0 Pronounced deposition of connective tissue in lung 0 Adenomatous proliferation of bronchial epithelium 0 7 OJ 0 *J W if 3 tn < 5 5 0 0 0 0 5 3 0 0 0 0 0 8 B___________13 13 m H 0 a0J u 0) if e o 3 u< 55 91 0 40J 4hJi m w e 0 9) u< 55 05 20 30 00 30 05 03 00 00 00 00 00 05 10 50 00 12 04 00 00 00 u0 00 00 Data listed as number of rats in each group shewing the various observations. 25____ 25 0 u *o n 4J G O U * < 44 04 00 00 02 00 04 01 03 01 01 01 01 S T 0 0 0 I^76 407150 TABLE 4 SUMMARY3 OF TUMOR INCIDENCE IN GROUPS OF MALE RATS USED IN INHALATION STUDY WITH ASBESTOS (Or Used as Controls for the Study) Control Number of Rats Examined Number of Rats with Tumors Number oi Tumors Mean Number of Tumors/Rats with Tumors 50 19 23 1.2 Lung 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 Subcutaneous Neurofibroma Carcinoma Fibroma Fibroadenoma 4/50 0/50 0/50 3/50 1/50 Pancreas Adenoma Islet cell 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 STOOO1478 aA record of observations on individual rats is contained in Appendixes A and B. bOne rat was missing. CFifteen of the 49 rats were killed for interim examinations by the 13th month bringing the actual incidence of pulmonary tumors up to 6 of 34. `Significantly increased as compared to controls when analyzed by Fisher's Exact Probability Test, p<0.05. 407151 ST000I479 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) Group '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 Zymball Gland Carcinoma 1/50 Small Intestine Mucinous adenocarcinoma 0/50 Testes Interstitial cell tumor 0/50 Intraabdomina1 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 1/4 9 1/49 A record of observations on individual rats is contained in Appendixes A and B. bOne rat was missing. CFifteen of the 49 rats were killed for interim examinations 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. 407152 (0 Jw 3 .J 03 & o JE u to Q to 22 o s to Du M 03 2 to o< OS X (14 uo * to >4 E-| WZ w u E-i MZ zo rj VO 00 c\ d) as o U OOO rC 4 dl ooo U I-I tfP i 1 U 0) oo CM in H 4-1 m D ai as oo o o oo di r4 4J i-4 H O <N rl r--1 o olo o df> 1 to H o\ o o fS U o o (N O a .-f U Id U o o to 4-1 a> d> u to 0) iH V> <N XT c d) 4-1 <r O ia-)i > -H d) 4-1 ooO * a> d) O 44 os to dP o o o 1 a) >< os 14 r- 00 CO mmm -C o o o u O o -i SI 1--4 d) id r--i -f-i in m rsi 4-) 4J r in <N <D 0 <N r-4 o to op O o to >- to S-l Ooo O SZ I-I U O * d) CaHU Qi3n to O' <T> r~ o *oJ -Hc aP O o o to kl a) a> xi > Ooo to O to 3 1 It] c n o H -c O4 id M O U -U U d) O4 to c o H to to H a> o H 4OJ It] 4-1 O to 4-* i--I 3 03 to 3 d) 4-1 H to OH --HC tl4 4-1 GO CD CD CD CO CD 407153 Rubber Stopper l $10001481 Fig. 1. A schematic diagram of the air elutriaror 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. 407154 i .i ) >. i i> i i I .} -i ST000I 482 . 2. Scanning electron micrograph of asbestos dusts retained on filter 2470x (2.5 cm=10y) 407155 Body Weights, Grams ST000 Hi 83 Figure 3 MEAN BODY WEIGHTS OF MALE RATS AND HAMSTERS EXPOSED TO ASBESTOS DUSTS OR MAINTAINED AS CONTROLS 407156 Figure 4 CUMULATIVE PERCENT M O R T A LIT Y OF MALE RATS EXPOSED TO ASBESTOS DUSTS AND THOSE M AINTAINED AS CONTROLS -co5 5 co o3 A <0 c0) XlL LU 407157 OO --H CD CD CD CO APPENDIX A 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 72-105 60 Gross: Killed; moribund, due to traumatic injury to upper incisors and overgrowth of lower incisors; decreased adipose tissue. Microscopic: Odontitis, peridontitis and gingivitis secondary to trauma; focal interstitial pneumonitis; focal hepatocellular degeneration and Inf lanzna cion. 72-146 109 Gross: Killed; cyst in the region of the brain and pituitary, 1 cm in diameter. 72-134 72-115 186 409 Microscopic: Proliferating cystic structure in region of ventral aspect of brain and pituitary (craniopharyngioma). Gross: Killed; large subcutaneous nodular structure weighing 169.5 grams, consisting of semi-gelai^ious tissue with non-flbrous areas; generalized anemia. Microscopic: Subcutaneous fibroma; Increased spleg^c hemopoiesis; increased hemopoiesis in liver, } Gross: Killed; large subcutaneous mass weighing 134.5 grams; heart dilated. CO cn Microscopic: Subcutaneous fibroma; slight chronic nephropathy; myocardial degeneration; intestinal nematodlasis. 72-131 409 Gross: Killed; excessive adipose tissue; edema of testes and scrotal region. Microscopic: Vacuolation of adrenal cortical cells; pituitary adenoma; focal pancreatic atrophy and fibrosis; hyperplasia of pancreatic islet; slight chronic nephropathy; isolated focus of inflam matory cells in lung. 407158 APPENDIX A (Continued) GROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING MAINTAINED AS CONTROLS FOR INHALATION STUDY WITH Af jESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at time of Death ______________ Gross and Microscopic Observations 72-148 413 Gross: Killed; moribund; over 130 ml of serosanguineous fluid in abdomen; highly lobulated dense white mass incorporating stomach, small and large intestines, and mesentery; metastatic trans plantations on the diaphragm and peritoneal cavity. Microscopic: Intraabdominal sarcoma, with extension via abdominal region; moderate chronic nephropathy; increased splenic hemopoiesis; scattering of alveolar macrophages in lungs. 72-149 420 Gross: Died; advanced autolysis; multiple nodular areas in the 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; hepatocellular hyper plastic nodule. 72-139 423 Gross: Killed; enlarged lobes of prostate containing green-yellow exudate. Microscopic: Moderate chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; intestinal nematodiasis; increased splenic hemo poiesis; Increased reticuloendothelial proliferation in spleen; hepatic lipidosis; diffuse suppurative inflammation adjacent to urinary bladder and sex glands. t/1 --I CD CD CD 407SO -`=" co cn 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 72-108 456 Gross: Died; advanced autolysis; enlarged, pitted, mottled surface appearance of kidneys: gastro intestinal contents consisting of dark black material, suggestive of uremia. and Microscopic: Pronounced chronic nephropathy with mineralization; secondary parathyroid hyperplasia; myocardial degeneration with mineralization; aortic mineralization. 72-137 72-106 485 504 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. Gross: Died; depleted adipose tissue; enlarged, dilated kidneys; aorta dilated; darkening of lungs possibly post-mortem; testes decreased In size. Microscopic: Pronounced chronic nephropathy with mineralization; myocardial degeneration with mineralization; degeneration of myocardial vessels; testicular atrophy; aortic minerali zation; lung mineralization. ST000I 487 407160 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 time of Death ________________Gross and Microscopic Observations 72-109 513 Gross: Killed; overgrown incisors and necrotic area of hard palate; pale, pitted kidneys with numerous cystic spaces. Microscopic: Moderate chronic nephropathy; degen eration of myocardial vessels; increased splenic hemopoiesis; secondary parathyroid hyperplasia; periarteritis; peribronchiolar lymphoid aggre gates; cne focus of alveolar fibrosis and cholesterol cleft formation. 72-113 529 Gross: Died; moderate autolysis; mottled surface of kidneys. Microscopic: Pronounced chronic nephropathy; hepatic congestion; increased splenic hemopoiesis; myo cardial degeneration; vacuolation of adrenal cortical cells; periarteritis, dystrophic miner alized focus in lung. 72-111 536 Gross: Died; subcutaneous nodule weighing 7.4 gm, 2 cm in diameter; lungs appear darkened; en larged kidneys with mottled surface appearance. Microscopic: Pronounced chronic nephropathy; myocardial deneneration; degeneration of myo cardial blood vessels; increased splenic hemo^ polesls; hepatic congestion; subcutaneous fil adenoma. 98*110001 407161 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 72-112 543 Gross: Died; moderate autolysis; overgrown incisor; enlarged kidneys, mottled surface appearance; dilated renal pelvis. 72-118 547 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. ^Si --J CD Gross: Killed; moribund; corneal opacity; subcutanSoUs nonfibrous mass adjacent to ear canal. CD Microscopic: Zymbal gland carcinoma; moderate chrotelc nephropathy; increased pigment in spleen; vacuolation of adrenal cortical cells. ^ 72-120 556 Gross: Died; moderate autolysis; enlarged pituitary; excessive mammary development. Microscopic: Pituitary adenoma; pronounced chronic nephropathy with mineralization; myocardial de generation; degeneration of myocardial vessels; aortic mineralization; isolated foci of in flammatory cells In lungs; subcutaneous galactocele. 72-121 556 Gross: Died; kidneys pitted and roughened; inelastic aorta; nodular areas in pancreas. Microscopic: Moderate chronic nephropathy; myocardial degeneration; aortic mineralization; no change in pancreatic tissue examined. 407162 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 time of Death Gross and Microscopic Observations 72-123 604 Gross: Killed; large subcutaneous 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, pitted kidneys with mottled surface; hydrothorax; 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 within alveoli of lung mineralization of lungs. 06*11OO'OlS 407163 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 Nunber Days on Test ac time of Death ________________ Gross and Microscopic Observations 72-129 634 Gross: Died; left corneal opacity; enlarged, pale, mottled surface of kidneys vith cystic structures; decreased adipose tissue; aorta distended vith loss of elasticity; testes decreased in .size and flaccid; stomach contained red material; mesenteric lymph nodes tortuous and nodular. Microscopic: Pronounced chronic nephropathy; myocardial degeneration and inflammation; periarteritis; testicular atrophy; adrenal cortical hyperplastic nodule; vacuolatlon of adrenal cortical cells; secondary parathyroid hyperplasia; increased reticulo endothelial proliferation in spleen; peribronchiolar lymphoid aggregates; edema of lymph nodes; aortic mlrerallzation. 72-128 72-132 641 652 Gross: Died; slight autolysis; darkening of lungs; enlarged pituitary (assumed pituitary adenoma.). CO _J Microscopic: Moderate chronic nephropathy; myocardial* degeneration; postmortem congestion of lungs; pituitary unavailable for examination. Gross: Killed; moribund; bilateral, partial cornea^ n opacity; mottled surface appearance of lungs; ___ enlarged, pitted, mottled kidneys with cystic structures; mesenteric aneurysm; testes decreased in si.e and flaccid; anemic appearance of tissues. Microscopic: Pronounced chronic nephropathy; Increased splenic hemopoiesis; vacuolation of adrenal cortical cells; periarteritis; myocardial degeneration; aortic mineralization; increased alveolar macro phages; proteinaceous material and fibrosis of alveolar walls; thoracic lymph nodes edematous; testicular atrophy. 407164 APPENDIX A (Continued) CROSS AND MICROSCOPIC OBSERVATIONS ON PATS 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 72-133 665 Gross: Died; moderate autolysis; enlarged prostate; mesenteric vessels dilated; pitted, cystic cortex of kidneys; enlarged parathyroids; gastric mucosa mineralization; testes decreased in size and flaccid. Microscopic: Periarteritis with thrombosis; pronounced chronic nephropathy; testicular atrophy, myocardial degeneration; secondary parathyroid hyperplasia; vacuolation of adrenal cortical cells; suppurative prostatitis; increased alveolar macrophages (heart failure cells) in lung. 72-135 72-136 697 728 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. OO Microscopic: Moderate chronic nephropathy; increased0 splenic hemopoiesis; increased pigment in spleenV~ myocardial degeneration; adrenal cortical hyper-0 plastic nodule; adrenal hematocyst; edematous _ mesenteric lymph nodes; pituitary unavailable fo?" examination. io Gross: Died; enlarged, pale, mottled 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 atrophy; aortic mineralization; atrial thrombosis; increased alveolar macrophages within alveoli and focal fibrosis of alveoli. 407165 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 time of Death ________________ Gross and Microscopic Observations 72-143 739 Gross: Killed; moribund; emaciated; 25-30 ml serosanguineous fluid in peritoneal cavity; enlarged, pale kidneys with cystic structures; multiple coalescing pale foci in liver; tough focal capsular 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 parathyroid 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 pheochromocytoma; hepatic cyst; pancreatic adenoma. 72-145 756 Gross: Died; advanced autolysis; enlarged, mottled surfaces of kidneys; enlarged parathyroid glands. Microscopic: Pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; secondary parathyroid hyperplasia; testicular atrophy; periarteritis; aortic mineralization; intra-alveolar aggregates of exfoliated alveolar macrophages (heart failure cells). GO --I CD CD CD CD CD 407166 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 72-104 Days on Test at time of Death ________________Gross and Microscopic Observations 870 Gross: Killed; corneal opacity; enlarged parathyroid glands; enlarged, mettled surface of kidneys; enlarged right adrenal (10-15 x normal), dark and hemorrhagic in color; two nodule ' 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 pheochromocytoma; 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 content of accessory sex glands; increased number of reticuloendothelial cells in liver; subpleural aggregates of alveolar macrophages containing hematogenous pigment. STOOOI494 407167 APPENDIX A (ContinucdO 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 time 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 parathyroids; hydrothorax; heart distended and pale; left atrial thrombosis. Microscopic: Pronounced chronic nephropathy with cyst formation; chickening of gastric mucosa with dis tended gastric pics; pulmonary exfoliation of alveolar macrophages; focal fibrosis and edema of lymph nodes; degeneration of myocardial vessels; myoca.dial degeneration; atrial thrombosis; diffuse keratitis; aortic mineralization; secondary 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 cell adenoma; increased size of pancreatic islet hepatic cyst formation, diffuse hepatocellular cytoplasmic vacuolation, lipidos bile duct hyperplasia, focal necrosis and in flammation and microcyst formation. i>T000 I 495 407168 Figure 5 CUMULATIVE PERCENT M O R TAILITY OF MALE HAMSTERS EXPOSED TO ASBESTOS DUSTS AND THOSE MAINTAINED AS CONTROLS O S e3 a 2co> E I Uxl CO --I CD CD CD er CD (JTk 407169 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 time of Death ______________Cross and Microscooic Observations 72-104 870 Gross: Killed; corneal opacity; enlarged parathyroid glands; enlarged, mettled surface of kidneys; enlarged right adrenal (10-15 x normal) , dark and hemorrhagic In color; tvo 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 pheochromocytoma; 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 content of accessory sex glands; increased number of reticuloendothelial cells in liver; subpleural aggregates of alveolar macrophages containing hematogenous pigment. STOOD I 497 407170 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 Microscopic Observations 72-198 200 Gross: Killed; moribund due to traumatic injury to lower inrtsor teeth and resultant hemorrhage; focal white areas In lungs, suggestive of aggregates of alveolar macrophages. 4 72-180 72-154 260 409 Microscopic: Lungs contained aggregates of dust parti cles, sometimes within cytoplasm of alveolar macro phages; adajcent alveolar walls showing minimal hypercellularity; aggregates of dust particles in thoracic lymph nodes; slight chronic nephropathy. Gross: Died; advanced autolysis; Increased fluidity of Intestinal contents; focal white areas in lungs suggestive of aggregates of macrophages. Microscopic: Pulmonary aggregates of particle-laden macrophages; aortic mineralization; pronounced chronic nephropathy; myocardial degeneration; testicular atrophy; decreased content of accessory sex glands. Cross: Killed; Subcutaneous cyst formation within perineal area. Microscopic: Subcutaneous cyst formation and granulom atous reaction; slight chronic nephropathy; myo cardial degeneration; isolated foci of hypercellu larity and thickening of alveolar cells adjacent to aggregates of dust particles, frequently within alveolar macrophages of the lungs; equivocal to very slight increase in fibrogenic tissues within thick ened alveoli; pulmonary interstium and thoracic lymph nodes containing aggregates of dust parti but not accompanied by any fibrogenic response. 8 6 * ll0 0 0 B 407171 APPENDIX B (Continued) CROSS AND MICROSCOPIC OBSERVATIONS ON RATS DYING WHILE BEING USED IN INHALATION STUDY WITH ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number 72-152 Days on Test at Time of Death______ 431 Gross and Microscopic Observations UD Gross: Killed; corneal opacity; overgrown incisorsjLO Isolated subpleural circumscribed white foci in the lungs, 1 mm in diameter, which disappeared upon inflation with formalin; mesenteric tissue thickened in one area, 3 cm in diameter. Microscopic: pulmonary reaction similar to 72-154; focal aggregates of inflammatory cells in liver; increased splenic hemopoiesis; slight chronic nephropathy; mesenteric fat necrosis and inflamma tion; pituitary microcyst formation; vacuolation of adrenal cortical cells; diffuse keratitis. 72-169 477 Gross: Killed; moribund; enlarged, mottled kidneys with cystic structures; testes decreased in size; mesenteric vessels dilated and sclerotic; diffuse darkened areas in lungs, one of which appeared firm and nodular upon palpation; thoracic lymph nodes enlarged and darkened. Microscopic: pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; hepatic lipidosis; periarteritis, testicular atrophy; Increased splenic hemopoiesis; secondary parathyroid hyperplasia; adrenal pheochromocytoma; vacuolation of adrenal cortical cells; aortic mineralization; pulmonary papillary adenocarcinoma; pulmonary exfo liation of alveolar macrophages (heart failure cells) plus edema, hemorrhage and reaction as described for 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 chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; occlusive hemorrhagic rhinitis; same pulmonary reaction as 72-154. 407172 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. 72-153 529 Microscopic: Prounounced chronic nephropathy; pulmonary reaction similar to 72-154; aortic mineralization; increased pigment in spleen; myocardial degeneration degeneration of myocardial vessels; secondary parathyroid hyperplasia; vacuolation of adrenal cortical cells; periarteritis. Gross: Died; moderate autolysis; kidneys roughened on surface. 72-155 532 Microscopic: Pronounced chronic nephropathy; secondary parathyroid hyperplasia; cystic retention of material within thyroid; myocardial degeneration; periarteritis, aortic mineralization; pulmonary reaction like 72-154. Gross: Died; advanced autolysis; fluidity of contents of distal intestinal tract, dark red In color; pale, pitted kidneys, with mottled surface appear ance . :!icroscopic: Pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; periarteritis, testicular atrophy; pulmonary reaction similar to but less than 72-154. 407173 oo CD CD CD cn CD CD 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 72-137 554 Gross: Died; advanced autolysis; hydrothorax; lungs filled with fluid. 72-160 581 Microscopic: Pronounced chronic nephropathy; myocardial degeneration; degeneration of myocardial vessels; periarteritis; vacuolation of cortical cells; focal hepatic fibrosis; epidermoid carcinoma of lung with met'stasis to thoracic lymph node; also pul monary reaction like 72-164 plus exfoliation of macrophages (heart failure cells). Gross: Died; moderate autolysis; one subpleural cir cumscribed pale foci in lung; enlarged, pale kidneys. 72-162 60? Microscopic: Pronounced chronic nephropathy; myocardial degeneration; vacuolation of cortical cells; pulmonary reaction similar to 72-154. Gross: Killed; moribund; darkened areas in lungs firm on palpation; thoracic lymph nodes containing red pigment; kidneys pitted with mottled surface appear ance and cystic areas; enlarged parathyroids; aortic arch dilated and inelastic. Microscopic: Pulmonary reaction similar to 72-154 plus focal alveolar hyperplasia and fibrosis; rvonounced chronic nephropathy; increased splenic hemopoiesis; secondary parathyroid hyperplasia; periarteritis; testicular atrophy; aortic mineralization; vacuo lation of cortical cells. GO --i CD CD CD cn CD 407174 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 Nuu.ber Days rn Test at Time of Death ______________ Gross and Microscopic Observations 72-163 625 Gross: Died; hydrothorax; several subpleural circum scribed pale foci up to 4 mm 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 nephropathy; periarteritis. 72-165 630 Gross: Died; enlarged, pitted kidneys; enlarged sublumbar 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; edena 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 nephropathy, 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. ST000I 502 407175 APPENDIX B (Continued; GROSS AND MICROSCOPIC OBSERVATION'S ON RATS DYING WHILE BEING USED IN INHALATION STUDY Will! ASBESTOS (Initial Group Size 50 Male Rats) Pathology Number Days on Test at Time 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; gastrointestinal tract darkened; gastric wall chickened; aorta dilated and Inelastic; no adipose tissue; testes decreased in 6ize. Microscopic: Lung reaction like 72-154 plus focal fibrosis and alveolar hyperplasia; aortic minerali zation; secondary parathyroid hyperplasia; myo cardial degeneration and fibrosis; pronounced chronic nephropathy; periarteritis. 72-171 658 Gross: Died; enlarged kidneys with cysts and mottled surface; enlarged parathyroid; enlarged pituitary dark in appearance; turbinates congested or hemorrhagic. Microscopic: Focal pulmonary lung fibrosis and alveolar hyperplasia; also reaction like 72-154; pronounced chronic nephropathy; hepatic portal inflammation; myocardial degeneration; pituitary adenoma; parathyroid not examined. C/7 H CD CD CD cn CD CO 407176 APPENDIX B (Continued) CROSS 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 72-172 661 Gross: Killed; two subpleural circumscribed gray-whice foci in lungs 3 mm 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; moderate chronic nephropathy; myocardial degeneration; increased splenic hemopoiesis; periarteritis; secondary parathyroid hyperplasia; adrenal pheochromocvtoma; mucinous adenocarcinoma 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; tesres decreased In size. Microscopic: Pronounced chronic nephropathy; myocardial degeneration; vacuolation of cortical cells; adrenal hematocyst; periarteritis; left atrial thrombosis; secondary parathyroid hyperplasia; aortic mineralization; pulmonary reaction like 72-164 plus exfoliation of alveolar macrophages. ST000I 50U 407177 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 72-190 697 697 Gross: Killed; moribund; multi-loculated hemorrhagic mass within mesentery, 10 cm in diameter; firm ncdular structure in pancreatic area of mesentery; enlarged pale spleen; liver pale; aorta with loss of elasticity. Microscopic: Intraabdominal malignant schwannoma; aortic mineralization; pulmonary reaction similar to but less than 72-164; increased splenic hemopoiesis; slight chronic nephropathy; myocardial degeneration; focal aggregates of reticuloendothelial cells in liver; pancreatic islet cell adenoma. Gross: Died; subcutaneous multi-nodular structure weighing 21 grams; hydrothorax; 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; aortic mineralization; pronounced chronic nephropathy; periarteritis; myocardial degeneration; degeneration of myo cardial vessels; focal hepatic portal fibrosis; subcutaneous fibroma; adrenal pheochromocytoma; secondary parathyroid hyperplasia; edema and congestion of lymph nodes. ST000I505 407178 0001507 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 72-185 724 Gross: Killed; moribund; scattered subpleural circum scribed pale foci in lungs up to 2 mm in diameter; aorta dilated with loss 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 cell 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 multiple cystic areas; right 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; hydrothorax; 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 cm in diameter. Microscopic: Pulmonary reaction similar to 72-154, plus increased exfoliated alveolar macrophages and focal alveolar fibrosis; aortic mineralization; adrenal hematocyst and vacuolation of cortical cells; peri arteritis and thrombosis; pronounced chronic ne phropathy with cyst formations and fetalization of epithelium lining tubules; testicular atrophy; hepatic lipidosis and extramedullary hemopoiesis; granulomatous colitis; secondary parathyroid hyper plasia; myocardial degeneration; atrial thrombosis; pituitary adenoma. 407179 APPENDIX B (Continued) CROSS 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 72-200 732 Gross: Killed; moribund; enlargen, hemorrhagic pituitary: enlarged, pale prostate, with purulent material: enlarged, pale kidneys; enlarged parathyroids. Microscopic: Pituitary adenoma; aortic mineralization; vacuolation of adrenal cortical cells; periarteritis; secondary parathyroid hyperplasia; myocardial de generation and fibrosis; moderate chronic nephropathy with mineralization; focal biliary hyperplasia in liver; increased splenic hemopoiesis; pulmonary reac-ion like 72-154; prostate not examined. 72-192 812 Gross: Died; enlarged, dark kidneys with scattered pale foci; enlarged liver; 2 subpleural circum 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 Gross: Died; subcutaneous mass 2.5 cm in diameter; hydrothorax; left atrial thrombosis; enlarged kid neys mottled in appearance; one thyroid enlarged and da'k; 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 Jibrosis; subcut. eous carcinoma; pituitary not examined (adenoma:); diffuse keratitis. ST000 I 508 407180 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 72-196 843 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, mottled in appearance; subcapsular hepatic pale pinpoint foci, 1 mm in diameter; hydrothorax; left atrial thrombosis; enlarged thoracic lymph nodes. Microscopic: Pulmonary reaction like 72-154, plus more alveolar macrophage exfoliation and focal fibrosis of alveoli; aortic mineralization; focal amyloidosis and reticuloendothelial cell proliferation in liver; pronounced chronic nephropathy; myocardial degeneration; atrial thrombosis; suppurative pros tatitis and urocystitis; vacuolation of cortical cells and adrenal hematocyst; thyroid adenoma; testicular atrophy; interstitial cell tumor of testicle. 72-197 72-199 865 - Gross: Died; corneal opacity; nodular area in left lung; firm on palpation; firm nodular structure on primary bronchus, 8 mm in diameter; liver mottled in appearance with scattered subcapsular pale foci up to 1 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; pronounced chronic nephropathy; thyroid adenoma; secondary parathyroid hyperplasia; vacuolation of adrenal cortical cells; splenic congestion; myocardial de generation; hepatic congestion and bile duct hyperplasia; aortic mineralization. oo No animal available for examination, probably due to | cannibalism or loss of rat. CD CD cn CD k> 407181 GROSS AND HISTOPATHOLOGIC OBSERVATIONS ON NON-PULMONARY TISSU ES OF MALE RATS K IL L E D AT 7 , 8 OR 13 MONTH TIM E INTERVALS DURING IN H ALATIO N STUDY W ITH ASBESTOS C ontrol A s b e s to s C ontrol A sbestos C ontrol A sbestos D ata lis te d as number o f r a ts in each group show ing th e v a rio u s o b s e rv a tio n s . 0ISI0001S SBBJDUCd uj luomSyd jo uoxqcxnuinooc ICDoj OO OO r-t O 8B3J3UBd jo sjsojqjj XCI3T3SJ93UJ xB3i OO O O f-t o 3q8xxs *sjesjodornsq opuoxds pasBaaouj o O O O CO CM uo73BnjmBX3U"F pur uox3Ea3U3Sap XBjpjBOo/Cm xB3i 0-4 f-c O CM r-c ssqXooqrdoq 33BJ3U383P 03 3U33CfpB SXX33 Xj03BTnmrx3UX jo s33b83j88b p33cxosj O O r-< O l-f CM Bxscxpo3cm3u xuj3S33ux O O O > <0 ro a o 33Ba3pom `Xqsedojqdau ojuojqo o o o o -< o Xu <0 G. uo 3q8jx8 `Xq3Bdoaqd3U ojuoaqQ 0 0 0 0 in 0 >4 X BjAXsd XBU3a UI s3TSod3p p3zxxB-*3UIW r-4 O -* 0 0 0 xBU3J UTssxnqn3 S3Jsod3p p3ZjxB;t3UTH CM rH 0 0 0 0 Xbupj-jj uj s33b83j88b XT33 Xao3BnmBXJUi CM O 0 0 *n m xBU3Hsuoj3Bnuoj 3sbo aBxnqns m < <r m m Jinojvo pup sjAxad tbusj jo uoj3E3BX7(I -t 0 O O 0 0 sXaupj^ JO 33BJjnS p3X330H 0 0 CN| fH H H q3333 jobjouj uaoj8j3ao 0 0 O O O *H 3sXoo3rtaaq XBU3JPV 0 0 O O -4 O 83t?R 30 asqnmn m m m m m m dnojQ 3u3m3ea.11 XBAJ33UX 3QJX r-* 00 00 HH 407182 -CIO SS akd histopathologic observations o k non fulhohart tis s u e s o r MALE RATS K IL L E D AT 25 MONTH T IM E INTERVAL DURINC IN H A LA T IO N STUDY W ITH ASBESTOS Data, lis te d aa number o f ra ts In ascii group showing the v a rio u s observations. M W Tine Interval in Months #tor Pf00Ni Treatment Croup 0 Number of Rats Examined K Corneal opacity Ul Pale, enlarged and roughened kidney Cyst In kidney Increase In size of adrenal gland w ** Multiple pale aubcapsular foci In liver M M Increased prominence of parachyrolda o Emaciation, nasal porphyrins Mineralization of gastric wall ** M Decreased size of testes and sex glands o o tor n 3 n O Focal reddened area on pituitary o Contraction of left hock jolot o o Dilated renal pelvis o o Sucutaneous abscess o Mass adjacent to urinary bladder o o Degeneration of myocardial blood vessels K Myocardial degeneration. Inflanmatlon and fibrosis o M Decreased spermatogenesis k) Focal hepatic degeneration and inflassaatlon o Chronic nephropathy, moderate U* a* Chronic nephropathy, pronounced u O* Mesenteric, testicular periarteritis w Mineralization of aorta and thoracic vesaelu u* Testicular atrophy Vecuolation of adrenal cortical cells Kl Secondary parathyroid hyperplasia - Adrenal pheochromocytoma - Thyroid adenoma o M Hyperplastic nodule of adrenal medulla X o ryov oopo* g a 3r O0m " KJ Increased heaatogcnouB pigment in spleen K) O Increased splenic hemopoiesis i-- Hyperplastic nodule of adrenal cortex K> Pituitary adenoma K> - Focus of vacuolated hepatocytes In liver Undifferentiated sarcoma adjacent to urinary bladder and urethra 407183 APPENDIX E INCIDENCE OF TUMORS IN CROUPS 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 pheochromocytoma Pancreatic Adenoma Adrenal pheochromocytoma Thyroid adenoma Pituitary adenoma Adrenal pheochromocytoma Pancreatic islet cell adenoma Pituitary adenoma Adrenal pheochromocytoma Days on Test at Time of Death of Rat With Tumor 109 186 409 409 413 420 485 536 547 556 604 631 641 697 739 739 742 742 742 806 806 870 870 Rat Number 72-146 72-134 72-115 72-131 72-148 72-149 72-137 72-111 72-118 72-120 72-123 72-127 72-128 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 rats dying during study or at the interim kills. ST0001512 407184 APPENDIX P 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 Pulmonary papillary adenocarcinoma Adrenal pheochromocytoma Pulmonary epidermoid carcinoma Pulmonary epidermoid carcinoma Subcutaneous neurofibroma Pituitary adenoma Adrenal pheochromocytoma Mucinous adenocarcinoma of small intestine Thyroid adenoma Intraabdominal 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 72-186 72-186 72-186 72-186 72-188 407185 CO H CD /WV O -- CJ1 CO ' -V <.r kVf APPENDIX F (Continued) INCIDENCE OF TUMORS IN CROUP 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 Undifferentiated sarcoma attached to urinary bladder Adrenal pheochromocytoma Pituitary adenoma Pituitary adenoma Thyroid adenoma Subcutaneous carcinoma Pituitary adenoma Thyroid adenoma Testicular interstitial cell tumor 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 ST000I5I4 407186 LO in -- o o o t- --r-- r /s M 1 Ma1 to i K> M K) on NO N> | K> %J* SO F NJ I K|> K> 1 K) to0 1 ft KJ N> ro K> 3* in in mO KJ on OB o F o 7<9 Z O ** ON X* a 0 N< tt ^. 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