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Final Report 1-1729.005 SIX WEEK REPEATED SMOKE INHALATION IN MICE TO COMPARE HISTOPATHOLOGY AND SHORT-TERM ENDPOINTS USING TEST (D6 and D10) AND REFERENCE CIGARETTES Final Report For Lorillard Research Center 420 English Street P.O. Box 21688 Greensboro, North Carolina 27420 September 25, 1985 By Microbiological Associates Inc. 5221 River Road Bethesda, Maryland 20816

Final Report I-1729.005 Table of Contents Page I. Data Page 3 II. Introduction 4 III. Purpose 6 IV. Reference and Test Cigarette Characterization 7 V. Test Description 9 VI. Materials and Methods 10 VII. Results and Discussion 16 A. General Toxicology 16 B. Clinical Chemistry and Hematology 19 VIII. Conclusions 21 IX. Figure and Tables 22 X. References 23 XI. Quality Assurance Statement 25 XII. Appendix A - Pathology Report 2

Final Report I-1729.005 I. DATA PAGE Cigarette Identity: Reference, Test Cigarettes D6 and D10 Dates Samples Received: June 9, 1982 (Reference) February 4,1983 (D6 and D10) Date Sample Returned: Retained in storage Initiation Date: March 24,1983 Completion Date: See Review Completed Date: Page 24 MA Experiment Number: 1-1729.005 MA Notebook Number: 1729.005 Archives Location: 5221 River Road, Bethesda, Maryland 20816 Sponsor: Lorillard Research Center 420 English Street P.O. Box 21688 Greensboro, North Carolina 27420 Authorized Representative: Harry Minnemeyer, Ph.D. Connie Stone, Ph.D. Testing Facility: Microbiological Associates Inc. 5221 River Road Bethesda, Maryland 20816 Lead Technician: Dj6vhd R. Dansie ~ Study Director: Rayny§nd M~-Bwid, Ph.D. 'r Z i S'j--- Da e 3

Final Report I-1729.005 II. INTRODUCTION Alterations in the morphology of the respiratory tract have been noted in animals exposed to whole cigarette smoke. The magnitude of the changes depend upon the species, the cigarette type, the smoke exposure regimen or dose, and the duration of the exposure. Recent reports have detailed procedures for standardized microscopic examination of the upper respiratory tract (Young, 1981) which complement the procedures for evaluation of the lower respiratory tract. These procedures permit visualization of the nasal passage, trachea, major bronchi, terminal airways and lung parenchyma (Dungworth et al, 1976). Thus a detailed evaluation will be performed on the following tissues from animals exposed to smoke for six weeks: 0 nasal cavity and nasal turbinates 0 oral cavity, larynx, and pharynx 0 trachea 0 lungs and bronchi 0 any other abnormal tissues In addition to the microscopic evaluation, the following 4 assays complement the histopathology endpoints and were performed on the same group of animals at the same time. These particular assays have been grouped together to most efficiently utilize the Walton Smoking Machine with 12 mice/machine. Clinical Chemistry: Changes in clinical chemistries can reflect generalized toxic effects to the lung, liver, kidney, heart or other organs by the alteration of specific blood enzyme levels (i.e., lactic dehydrogenase), blood glucose, blood urea nitrogen, ion or metal levels, and total protein levels. Hematology Assays: Hematologic changes can reflect the effects of a large number of chemicals or agents on the blood and blood-forming organs, either directly or indirectly. Decreased or increased numbers of erythrocytes or leukocytes in the peripheral blood can reflect diseased or toxic states. 4

Final Report I-1729.005 Metabolism - Microsomal Monooxygenase (MMO) Levels: Aryl hydrocarbon hydroxylase (AHH) is one of the microsomal monooxygenases and represents the enzymatic complex responsible for the metabolism of polycyclic hydrocarbons, such as benzo(a)pyrene (BaP), benz(a)anthracene, or 3-methylcholanthrene. It is present in most mammalian tissues and can be enhanced in animals by the administration of a variety of chemicals such as the polycyclic hydrocarbons themselves, 5,6-benzoflavone, or 2,3,7,8- tetrachlorodibenzo-p-dioxin (Kouri and Nebert, 1977). The enzyme is detected in a fluorometric assay which quantifies the conversion of BaP to 3-hydroxy-BaP. AHH levels will serve as internal markers in tissues for particulate deposition or redistribution after smoke exposure. Comparative levels of AHH in lung, liver, and kidney between cigarette types would allow differences in deposition, absorption, and redistribution to be observed. This may be particularly important if additives to the particulate phase involve AHH inducers. Cellular Proliferation: Cell turnover or replication can be estimated by either mitotic indices or incorporation of 3H-thymidine into DNA. These techniques are sensitive criteria indicating possible induced DNA or cellular damage, where no evidence of damage can be seen by histologic appearance (Lamb and Reid, 1968). Cigarette smoke has been shown to induce cellular proliferation in mice (Kanagalingam et al, 1982). Mitotic counts can be evaluated in the same slides (and animals) used for "general" histopathologic evaluations (Bertalanffy, 1964) and can be easily incorporated into these studies. 5

Final Report I-1729.005 III. PURPOSE The purpose of this study is to compare the response of animals exposed to smoke from the Reference cigarette with smoke from two test cigarettes using histopathology of the respiratory tract and selected short-term endpoints. 6

Final Report 1-1729.005 IV. CIGARETTE CHARACTERIZATION Approximately 2000 Reference cigarettes were received on June 9, 1982, and 1200 D6 test cigarettes, and 1200 D10 test cigarettes were received by Microbiological Associates Inc. on February 4, 1983. The Reference cigarette was assigned a registry number of 5, D6 a registry number of 14, and D10 a registry number of 15. Characterization of cigarette smoke was performed each week and included determination of resistance-to-draw (RTD), butt length, and wet total particulate matter (TPM) per cigarette. RTD was measured using a Filtrona Pressure Drop Tester (Cigarette Components UK Ltd., Wembley, England). Butt length was measured after burning cigarettes with 7 puffs/cigarette. TPM was collected onto Cambridge filter pads from each cigarette after burning on the Walton Horizontal Smoking Machine. The standardized conditions for burning cigarettes were the same as those used for exposing the animals (7 puffs). The following data were collected: Cigarette RTD Butt TPM Type (mm H20) Leygt~ (mg) mm Week 1 Reference 79.7 + 0.6 44.7 + 2.1 15.3 + 0.2 D6 101.7 + 6.0 47.3 + 2.3 13.8 + 0.4 D10 88.0 + 2.6 47.0 + 5.6 14.4 + 0.6 Week 2 Reference 80.0 + 1.4 44.0 + 4.2 14.9 + 0.1 D6 102.3 + 4.6 51.0 + 6.0 10.7 + 0.5 D10 95.5 + 10.6 49.0 + 2.8 12.3 + 0.1 Week 3 Reference 80.7 + 2.1 40.3 + 3.5 18.1 + 0.3 D6 95.3 + 11.0 47.7 + 2.5 13.0 + 0.3 D10 91.7 + 1.5 49.3 + 4.7 15.4 + 0.4 Continued... 7

Final Report I-1729.005 Cigarette RTD Butt Type (mm A20) Length T PM (mg) --------------------------------------------------------- Week 4 Reference 89.3 + 4.0 44.0 + 4.0 15.5 + 0.4 D6 101.5 + 2.1 45.5 + 0.7 15.3 + 0.1 D10 100.5 + 7.8 52.5 + 3.5 13.8 + 0.8 Week 5 Reference 83.5 + 0.1 40.0 + 5.7 14.5 + 0.2 D6 104.0 + 1.4 46.0 + 1.4 13.6 + 0.5 D10 95.5 + 7.8 47.5 + 0.7 16.7 + 2.8 Week 6 Reference 84.0 + 1.4 46.5 + 2.1 13.5 + 2.8 D6 103.5 + 0.7 45.5 + 0.7 15.8 + 0.4 D10 87.0 + 4.2 49.0 + 1.4 15.3 + 0.1 The Sponsor specified storage at room temperature and no expiration date was provided. For the purposes of this study, the test articles were stored in the original cellophane sealed boxes in a secured area at room temperature. 8

Final Report I-1729.005 V. TEST DESCRIPTION B6C3F1/Cr1BL mice were exposed to smoke from two test cigarettes and the Reference cigarette. The exposure regimen used (7 puffs, RSD1) was decreased from that used (8 puffs, RSD1) in the acute toxicity study (I-1729.003) to reduce toxicity and ensure adequate survival. At weekly intervals, mice were weighed and monitored for levels of carboxyhemoglobin (COHb). Three mice/group were sacrificed after 3 weeks exposure to determine microscopic appearance of the respiratory tract, clinical chemistry, and hematology. After 6 weeks exposure, 3 mice/group were sacrificed to determine microscopic appearance of the respiratory tract, clinical chemistry, and hematology; another 3 mice/group were sacrificed for respiratory tract histopathology and mitotic counts in the same respiratory tissues; and the remaining 3 mice/group were sacrificed to determine AHH levels in lung, liver, and kidney. 9

Final Report I-1729.005 VI. MATERIALS AND METHODS Materials Animals: B6C3F1/Cr1BL mice, females 8 weeks old Sendai vaccinated Charles River Breeding Farms Kingston, New York Smoke Generation Equipment: Walton Horizontal Smoking Machine (Walton) Stock-like neck holders Process and Instruments Corp. Brooklyn, New York Materials: Heparinized blood collecting tubes (250 ul) IL-CO-Oximeter Metofane anesthetic Methods Animals: Animals were obtained from a source monitored and known to be free of adventitious agents, vaccinated against Sendai virus and quarantined for at least 21 days. Stringent disease control procedures were followed during quarantine to assure the use of healthy animals. Mice were observed for signs of illness, unusual food and water consumption and other signs of poor health. The animals were 12 weeks old at initiation of test and were judged to be healthy prior to utilization. Animals were housed in an AAALAC-accredited facility with a controlled environment of 74 + 5 OF, 50 + 20% relative humidity, and a 12 hour light/dark cycle. Mice were housed 6 per cage in polycarbonate autoclavable cages with filter top cage lids. Corn-cob bedding was used and animals had free access to certified laboratory rodent chow which had been analyzed for environmental contaminants. Water and food were provided ad libitum, except during exposures on the smoking machines. 10

Final Report 1-1729.005 Treatment: Animals were ear tagged and randomly assigned to 4 groups of 12 animals each using the stratified sequence randomization technique. Groups of mice (12 per group) were exposed to smoke from two test cigarettes and the Reference cigarette using the following defined exposure regimen on the Walton Smoking Machine (Henry et al, 1980, Guerin et al, 1979): 10% (v/v) smoke concentration (-4 ug TPM/ml) 30 seconds smoke/30 seconds air/minute 7 puffs/exposure 9 exposures/day 63 puffs/day 7-8 minute rest periods between exposures 31.5 minutes total time/day exposed to smoke -130 minute/day exposure period This daily (5 days/week) exposure regimen over 6 weeks resulted in: 270 total exposures 1890 total puffs 30 days exposed Exposure controls were used which consisted of age-matched mice loaded onto the smoking machine, treated exactly as the test animals, the machines set and run but without cigarettes. This sham-control group was also treated for approximately -130 minutes. The following designates the disposition of the animals: Group Description 1 Reference Cigarette 2 Test Cigarette 1 (D6) 3 Test Cigarette 2 (D10) 4 Sham-Exposed Control Animals were weighed 24 hours prior to the first exposure and weekly thereafter during treatment. Blood carboxyhemoglobin (COHb) levels were monitored in 2 mice/group at weekly intervals. 11

Final Report I-1729.005 Animals were observed during the test period for any unusual persistent conditions. Sacrifices: Animals were weighed to the nearest tenth of a gram prior to necropsy. Animals were anesthetized with Metofane. Blood samples from 3 animals/group at 3 weeks and 6 weeks were collected for clinical chemistry and hematology. Animals were killed by exsanguination. Necropsy: The necropsy is defined as external examination, including body orifices, and gross examination and fixation in 10% formalin of all of the following tissues and organs: Spinal cord (thoracic) Brain Eye (R) Pituitary Trachea Larynx Esophagus Adrenals (R & L) Thyroids (R & L) (with parathyroids) Lungs & Bronchi (R & L) Kidneys (R & L) Liver Spleen Thymus Any gross lesions Salivary gland (R) Mandibular lymph node Mesenteric lymph node Sternum (bone marrow) Zymbal's gland (R) Heart Thigh muscle (R) Sciatic nerve (R) Stomach Small intestine Pancreas Large intestine Turbinates Sk in Urinary bladder Females (R) = Right (L) = Left Ovaries (R & L) Uterus Mammary gland Gross observations made during necropsy were recorded on an individual animal necropsy form containing the animal identification number, group number, compound administered, date of death, necropsy date, signature block, study number, and other pertinent information. In addition, all tissues taken at necropsy were recorded on the necropsy form. Jars containing animal tissues were labelled with species, animal number, study number, date of necropsy and technician initials. 12

Final Report I-1729.005 The trachea was transected midway between lung and tongue, and the heart, thymus and extraneous tissues were carefully removed. The lung was weighed to the nearest milligram, and perfused with fixative via the trachea to approximate the normal expanded size. Histology: 0 Lungs - The preferred planes of section for rodents' lungs are illustrated in Dungworth et al, 1976. These were sections from the hilus along the axis of major airways for the cranial, middle and caudal lobes of the right lung and longitudinally through the main stem bronchus of the left lobe. Sections in this plane for each lobe revealed bronchial and acinar orientations of lesions much more readily. All of these blocks can be sectioned whole for microscopic examination. 0 Larynx - The larynx was transected from the trachea. Three transverse sections were taken from the larynx with the thyroid and parathyroids attached, one from the distal portion (vocal folds and caudal cricoid region), one from the middle portion (arytenoid projections and ventral pouch) and one from the proximal region (epiglottis). This will enable all major regions to be assessed. 0 Trachea and bronchial bifurcation - A block containing a longitudinal section of distal trachea (distal to larynx) was prepared to include the bronchial bifurcation. The bifurcation is a site sensitive to injury in inhalation studies because it is the major location where air flow changes direction and particles are likely to impinge. Only the respiratory tract organs were processed for histopathology at this time. Lung tissues were embedded in glycol methacrylate and sectioned at 2.5 microns. The remaining fixed tissues are being held until data are analyzed. Final disposition will be determined after consultation with the Sponsor. Blood Collections: Blood collections were made from retro-orbital sinus puncture or from anesthetized animals by cardiac puncture after access was gained by incision. Approximately 0.3 ml blood was used for hematology with potassium ethylenediaminetetraacetate (K-EDTA). K- EDTA was used as the anticoagulant since it does not adversely affect the shape of the red blood cells. Blood smears were prepared from a drop of fresh blood placed directly on a glass slide, spread, air dried and fixed in methanol. Blood for clinical chemistry was allowed to clot and serum separated from the cells. 13

Final Report I-1729.005 Clinical Chemistry: The following clinical Glucose (serum) Blood Urea nitrogen (BUN) Creatinine Cholesterol Triglycerides Calcium Protein, total (serum) A/G Ratio Hematology: The following hematological Segmented neutrophils count Monocyte counts Platelet cell counts Red blood cell counts (RBC) Basophil counts Hematocrit value Mitotic Counts: Alkaline Phosphatase Lactic dehydrogenase (LDH) SGOT SGPT Uric-acid (serum) Phosphorous Albumin tests were performed: Lymphocyte counts Eosinophil counts Reticulocyte counts White blood cell counts (WBC) Hemoglobin concentration For determination of mitotic indices in lung tissues, immediately after the last smoke or sham exposure, 3 animals/group were injected with 0.1 mg colchicine per 100 g body weight (Lamb and Reid, 1968). Four hours later, animals were anesthetized with Metofane and killed by exsanguination. Clinical chemistry and hematology were not determined on these animals. Necropsies were performed as described above and respiratory tissues examined microscopically. The mitotic activity of the bronchial epithelial cells was assessed. The number of nuclei of bronchial epithelium in mitosis was expressed per 12 diameters of a microscopic field using the 40x objective. Approximately 1000 cells were observed in each specimen. AHH Levels: For determination of AHH levels, 6 hours after the last smoke or sham exposure, 3 animals from each group were sacrificed by CO asphyxiation and lungs, liver, and kidneys dissected away from ?he carcass and weighed to the nearest 0.1 mg. Microsomes were prepared according to Burke and Mayer (1974) and stored at -70°C until assayed. AHH activity was determined by fluorometric chemistry tests were performed: 14

Final Report 1-1729.005 measurement of the 3-hydroxy-BaP (3-OH-BaP) formed according to Kouri et al. (1976). AHH units are expressed as pmole 3-OH-BaP formedTi-n7mg microsomal protein. Evaluation of Test Results The survival in each group is presented, as are any unusual observations during or after exposure over the 6 weeks. Blood COHb levels from 2 mice per group at weekly intervals are also presented. Individual animal weights 24 hours prior to exposure and at weekly intervals after exposure during treatment are presented for each group. Individual animal pathologies on the respiratory tract from 9 animals per group are presented, 3 animals/group after 3 weeks and 6 animals/group after 6 weeks). Total erythrocyte counts, total leukocyte counts, hematocrit, hemoglobin, differential leukocyte counts, blood urea nitrogen, blood glucose and other clinical chemistries from 3 animals/group sacrificed at 3 weeks and 6 weeks are presented. After 6 weeks of treatment, the levels of AHH in lung, liver, and kidney tissues from 3 animals/group and the Mitotic Index (MI) in lung tissue from 3 animals/group were determined. Statistical analyses of all the data presented were performed using BMDP-79 Biomedical Computer Programs, P-series (Dixon and Brown, 1979). Analysis of variance (ANOVA) was performed comparing the equality of means between groups and an F-statistic calculated. In addition, Levene's test for the homogeneity of variances was calculated, as were Welch and Brown-Forsythe statistics which assume unequal variances. . The data were entered as a matrix and the group means for each variable (column) compared, e.g. body weights of all animals on test for 7 days. Because the program only accepts data in matrix form, values that are not recorded must be entered into the computer as "missing" as opposed to zero and are omitted from analysis. Significant differences between individual means were determined by calculating a least significant range using a Studentized Range Test modified for unequal sample sizes (Sokal & Rohlf, 1969). Criteria For Determination of Valid Tests: None of the sham-exposed animals should die or show signs of toxicity. COHb levels in the smoke-exposed groups should be statistically (p < 0.05) increased relative to the sham-exposed controls. 15

Final Report I-1729.005 VII. RESULTS AND DISCUSSION A. General Toxicology B6C3F1 mice were quarantined for at least 21 days prior to use, during which time their health status was evaluated by observation. The animals were 12 weeks old and were judged to be healthy prior to the initiation of this study. Groups of animals were exposed "nose-only" to whole cigarette smoke generated on a Walton Horizontal Smoking Machine. Test cigarettes D6 and D10, and the Reference cigarette, were evaluated together using the 7 puff Reference Smoke Dose 1 (7 puffs, RSD1). This change from 8 to 7 puffs was adopted after consultation with the Sponsor because the results of the acute toxicity study (See Final Report, 1-1729.003) indicated that 7 puffs contained an amount of TPM that is equivalent to what has been found with other test cigarettes. Weekly determinations of TPM from the Reference and test cigarettes were taken and are presented in Section IV. Cigarette Characterization. The results indicate that the TPM varied only about 10-20% over a 6 week period and were within historical limits. The smoke-exposed groups were compared to a sham-exposed (no cigarette) control group. The Reference, D6, and D10 cigarettes had been previously evaluated using an 8 puff RSD1 regimen in an acute toxicity study. The results of that acute toxicity study suggested that a reasonably high dose of smoke could be given with good survival predicted in longer periods of exposure (see Final Report (1-1729.003) provided a change from 8 to 7 puffs was affected. Smoke and sham-exposures were initiated on March 24, 1983, and completed on May 4, 1983. Animals were treated 5 days/week for a total of 30 exposure days. One animal from Group 3 (#1173, test cigarette D10) died during exposure on Day 12 after 63 exposures, and one animal from Group 4 (#1187, sham-exposed) died during exposure on Day 19. This sham-exposed animal was found dead at the end of exposure and had not displayed any signs of struggling or toxicity. An autopsy did not reveal any overt cause of death. It is presumed that this animal died of accidental suffocation that was unrelated to the exposure. Aside from these two cases, the 3 animals/group scheduled for sacrifice at 3 weeks on test were sacrificed on schedule, with the remaining 6 animals/group sacrificed as scheduled after 6 weeks (Table GT1). Those animals sacrificed after 3 weeks received the scheduled 135 exposures and 945 puffs over 15 treatment days. While animals sacrificed after 6 weeks received the scheduled 270 exposures and 1890 puffs. Survival to the 7 puff, RSD1 regimen for the Reference and D6 cigarette smoke was 100% and 92% for smoke from cigarette D10. Survival to sham exposure was also 92% because of one accidental death. 16

Final Report I-1729.005 Animals were observed daily during and immediately after exposure for any unusual responses to treatment. No unusual responses were observed. Mice exposed to smoke were lethargic, ataxic, and felt hypothermic. There was tearing of the eyes, trembling, and shallow breathing observed after exposure. In addition, mice that had been exposed to smoke from test cigarette D10 were unable to walk and kept a prone position. Sham-exposed mice appeared alert and active after exposure, with the exception that occasional tearing of the eyes and rhinorrhagia was observed. The COHb levels from 2 mice per group determined at weekly intervals are presented in Table GT2. The COHb values for the smoke-exposed groups ranged from between 21.8 to 59.8% COHb. The smoke-exposed mice had significantly higher COHb levels than did sham-exposed controls (Studentized Range Test, p < 0.05; Table GT2). The individual animal weights 24 hours prior to the first treatment and at weekly intervals thereafter are presented for each group in Table GT3. Individual body weights for animals from each group are also presented in histogram form in Tables GT4 - GT10. Statistical descriptions for each group, as well as an Analysis of Variance, may also be found in Tables GT4 - GT10. The group means for each weekly interval are presented in Figure 1. All animal weights were relatively stable throughout the study and Analysis of Variance indicated that the body weights of the sham- and smoke-exposed groups were statistically the same (Analysis of Variance, p > 0.05). The lung weights of mice sacrificed after 6 weeks of treatment are presented in Table GT11. Lungs were dissected away from the thoracic viscera as described in the Methods, and weighed before perfusion with formalin fixative. The mean and standard deviations are presented for 3 animals/group. Analysis of Variance did not indicate any significant differences (p > 0.05). The AHH activities in the lung, liver, and kidneys of mice exposed to smoke for 6 weeks are presented in Table GT12. No increases were observed in the pulmonary or renal AHH activities of smoke-exposed mice. The AHH activity in the liver of smoke-exposed mice was increased above that in sham-exposed controls, but the increases were not statistically significant (ANOVA, p > 0.05). These findings do not compare well to results obtained in previous studies, which have always indicated induced AHH activity in the lungs of animals exposed to cigarette smoke. The level of enzyme activity is also not as high as had been obtained previously. This may be the result of the method used to isolate microsomal enzymes. Instead of ultracentrifugation, calcium chloride precipitation was used to pellet the microsomal fraction of the cell homogenates. This procedure was done because it is more rapid and does not require an ultracentrifuge. Preliminary results using a positive control of microsomes from 17

Final Report I-1729.005 animals treated with 3-methylcholanthrene had indicated that this method would be satisfactory for subcellular isolation of microsomes from smoke-exposed animals. However, the enzymes prepared in this way may be more susceptible to denaturation during freezing and thawing. As a result, calcium chloride precipitation will no longer be used to prepare enzyme fractions. Bronchial and tracheal epithelium were evaluated for changes in cellular proliferation after final smoke or sham treatment at 6 weeks. Twelve randomly selected fields from lung and 5 randomly selected fields from tracheal sections of 3 animals/group were examined at high magnification (400x) for the presence of mitotic figures. These fields represented approximately 1000 bronchial or tracheal epithelial cells, respectively. Very few mitotic figures were found in either the smoke-exposed or sham-exposed groups (less than an average of 4 and 1 mitotic figures/1000 bronchial and tracheal epithelial cells, respectively). These data suggest that increased numbers of mitotic figures could not be associated with this dose exposure regimen during a 4 hour colchicine treatment period. 18

Final Report I-1729.005 B. Clinical Chemistry and Hematology Clinical chemistry and hematology values for individual animals are presented in Tables CCH1-CCH4 for animals sacrificed after 3 weeks on test, and in Tables CCH28-CCH31 for animals sacrificed after 6 weeks on test. After 3 weeks on test, a few significant changes in the clinical chemistries were observed. Triglycerides were elevated in mice exposed to smoke from the Reference and D6 cigarettes as compared to sham-exposed controls (Studentized Range Test, p < 0.05; Table CCH7), but not in mice exposed to smoke from cigarette D10. However, inasmuch as the differences observed are only slightly beyond normal values, and there is no concomitant change in glucose or cholesterol levels, the results are not believed to be biologically significant. Serum creatinine and BUN were also elevated in all smoke exposed groups in comparison to the sham-exposed mice (Studentized Range Test, p < 0.05; Tables CCH8-CCH9). Elevation in both of these parameters is probably indicative of a possible decrease in glomerular filtration in the kidney. All the remaining clinical chemistries were within normal limits. Smoke-exposed mice also had significantly increased numbers of leukocytes (Studentized Range Test, p < 0.05; Table CCH2O). This appears to be a general response to smoke because the differential count of leukocytes did not indicate any abnormal cell populations. All the remaining hematology values were within normal limits. After 6 weeks on test, mice exposed to smoke from test cigarettes D6 and D10 had elevated triglyceride levels when compared to sham-exposed controls (Studentized Range Test, p < 0.05; Table CCH34). This was not the case in mice exposed to smoke from the Reference cigarette. Although similar results were observed after 3 weeks of exposure to smoke, the results do not appear to be indicative of any specific pathologic condition. As had been observed after 3 weeks of exposure, BUN was elevated in all smoke exposed groups (Studentized Range Test, p < 0.05; Table CCH36). An elevation in BUN without a concomitant change in creatinine may be the result of any number of 00 physiological effects and is not necessarily indicative of a m pathologic condition (Coles, 1980). Thus, the biological ~ significance, if any, of the increase in BUN is not known. ~ A significant decrease in the A/G Ratio was observed in mice W that had been exposed to smoke from test cigarette D6 when compared to animals that were exposed to the Reference cigarette and to sham-exposed controls (Studentized Range Test, p < 0.05; Table CCH40). No differences were found among the other groups. 19

Final Report I-1729.005 Since the differences observed were within normal limits, the results are not believed to be biologically significant. No differences in the hematology values of mice on test for 6 weeks were observed. 20

Final Report I-1729.005 VIII. CONCLUSIONS The criteria for a valid test were satisfied in that no sham-exposed animals showed signs of toxicity and COHb levels for the survivors in smoke exposed groups were significantly greater than the sham-exposed group. Overall survival for mice on test for 6 weeks was 100% for those groups exposed to smoke from the Reference and D6 cigarettes, and 92% for animals exposed to smoke from test cigarette D10. Survival in the sham-exposed group was also 92%. However, because no signs of toxicity were observed in the sham-exposed group, the one death that occurred can be attributed to accidental suffocation. No unusual signs of toxicity were observed in mice exposed to test cigarette smoke. Body weight changes and lung weights were statistically the same for all groups. The AHH assay was found to be invalid for this 6 week study. The clinical chemistry of mice sacrificed after 3 weeks on test was somewhat altered from the control. However, the only notable changes were for creatinine and BUN, which were both elevated in mice exposed to smoke. Changes in these parameters usually indicate a decrease in glomerular filtration (GFR), caused by any number of physiological conditions. Inasmuch as the changes occurred in all smoke exposed animals, the cause does not appear to be test article related. Instead, a component of all cigarette smoke, such as nicotine which is known to have antidiuretic properties, may have caused the decrease in GFR. Leukocytes were also increased in smoke exposed animals after 3 weeks on test, but this may be a general response to the smoke and not test article related. After 6 weeks on test, increased triglyceride levels were observed in groups exposed to test cigarettes, and increased BUN levels were observed in all smoke exposed groups. The biological significance of these alterations is not clear. No differences were observed in the hematological values of animals sacrificed after 6 weeks. In general, the mice tolerated the smoke exposure well and no-unusual signs of toxicity were noted. Smoke related responses were observed in the lungs of female mice exposed to smoke 5 days each week for 3 and 6 weeks, respectively. The responses included macrophage infiltration of alveoli and lymphoid infiltration of the interstitial tissue between small airways and vessels and infrequently around pulmonary vessels. Grossly, lung discoloration was observed in most smoke-exposed mice, but not sham controls. Control mice were essentially free of gross and microscopic responses. No differences between reference smoke and test article smoke exposed groups could be discerned. 21

IX. Figure and Tables Final Report I-1729.005 22

FIGJRF. 1 I 23 22 BODY WEICRIT DATA FROP4 1-1729.005 Final Report 1-1729.005 0 4 a o N 19 18 5 10 15 20 25 30 35 40 DAYS ON TEST 000qjLsB?3

Final Report I-1729.005 Table GT1 Disposition of Animals Number of Animals Sacrificed/ Number of Animals Scheduled for Sacrifice Group - Cigarette Number of A on Tes nimals t 3 Weeks 6 Weeks 1-Reference 12 3/3 9/9 2-D6 12 3/3 9/9 3-DlO 12 2/3 a 9/9 4-Sham 12 2/3 b 9/9 -------------------------------------------------------------------- aAnimal #1173 died during exposure on Day 12. bAnimal #1187 died during exposure on Day 19.

Final Report 1-1729.005 Table GT2 Percent Carboxyhemoglobin Levels in Mice Days on Test a Group- 7 14 21 28 35 42 Mean + Standard Deviation ---------------------------------------------------------------------- b 1-Ref 40.7 45.8 45.2 21.8 30.5 34.3 36.3 + 8.6 35.2 51.6 33.3 34.4 24.8 37.5 2-D6 59.8 46.4 42.2 44.0 47.6 29.7 42.1 b + 8.6 49.8 40.0 42.8 39.2 34.9 29.3 3-D10 59.6 57.4 54.1 34.4 41.0 32.5 44.8 b,c + 12.1 57.6 54.4 46.7 22.4 34.7 43.1 4-Sham 2.2 1.4 2.2 2.7 2.7 1.3. 2.1 c + 0.5 1.7 2.3 1.3 2.2 2.6 2.1 aWithin 3-5 minutes after last exposure, 2 animals/group were bled from the retro-orbital sinus for COHb determination. bMean COHb values significantly higher (p < 0.05 Studentized Range Test) from sham-exposed group. COne animal from this group bled on successive weeks because of attrition in that group.

TABLE GT3 INDIVIDUAL BODY WEIGHTS OF MICE ON TEST Final Report I-1829.005 C A S E 1 3 4 5 6 7 8 9 NO. LABEL CIG BEFORE DAY7 DAY14 DAY21 DAY28 DAY3S DAY42 ---- -------- ---------- ---------- ---------- ---------- - -- ---------a ---------- ---------- 1 1145 1(Ref) 21.500 19.900 20.600 20.200 MISSING MISSING MISSING 2 1146 1 22.200 20.700 20.700 20.600 a MISSING MISSING MISSING 3 1147 1 21 20.700 19.300 20.400 MISSINGa MISSING MISSING 4 1148 1 20.400 19.700 20 20.700 20.700 20.400 22.400 5 1149 1 20.800 20.500 19.900 20.200 20.400 20.400 22.300 6 1150 1 22.200 20.400 20.500 21.200 20.900 21.300 22.200 7 1151 1 22.100 20.900 20.300 20.900 20 20.800 21.700 8 1152 1 21.600 20.500 20.500 20.700 20.300 20.600 21.400 9 1153 1 21.500 20.600 21.600 21.800 21.100 21.500 22.900 10 1154 1 21 19.200 19.600 21.300 19.600 20.300 21.300 11 1155 1 22.900 21.800 21.300 21.900 21.600 21.500 22.600 12 1156 1 21.800 21 21.100 22.100 20.600 21.600 22 13 1157 2(D6) 17.900 17.200 17.700 18.600 MISSINGa MISSING MISSING 14 1158 2 23.200 22 21.400 21.800 MISSINGa MISSING MISSING 15 1159 2 21.600 21.500 20.700 21.900 MISSINGa MISSING MISSING 16 1160 2 20.900 20.800 20.300 21.100 21.400 20.200 21.400 17 1161 2 20.700 20.700 20.500 21.100 20.800 20.200 21.400 18 1162 2 19.900 19.700 19.400 19.200 19.700 19.600 20 19 1163 2 20.900 20.900 20 20.200 20.300 20.400 21.800 20 1164 2 21.700 21.300 21.100 20.800 21 21 21.600 21 1165 2 20.200 19.700 20 20.300 19.600 20.200 21.300 22 1166 2 23.300 22.800 22.300 21.800 22.100 22.400 24 23 1167 2 21.600 20.600 21 21.400 21.600 21.800 21.500 24 1168 2 21.900 20.500 20.700 20.600 20.500 20.300 20.500 25 1169 3(D10) 21.200 20.600 20.600 21.400 MISSINGd MISSING MISSING 26 1170 3 20.700 19.700 19.600 20.800 MISSINGa MISSING MISSING 27 11.71 3 20.800 20.600 19.600 20.900 20.400 21 20.900 28 1172 3 22.100 20.600 20 21 21 21.200 20.700 29 1173 3 22.200 21.600 MISSING h MISSING MISSING MISSING MISSING 30 1174 3 20.200 19.800 20 20.100 20.700 21.100 20.900 31 1175 3 23.300 21.900 21.300 22.200 22.600 22.800 23 32 1176 3 20.500 20.600 20 20.500 20.500 20.800 21.800 33 1177 3 21.700 22.200 21 21.600 20.400 21.200 21.800 34 1178 3 22.900 21 21 20.500 21.800 20.600 21.200 35 1179 3 21.400 20.600 19.700 20.500 20.800 20.800 20.500 36 1180 3 21.200 21.400 21.700 22 21.800 21.900 23.300 37 1181 4 (Sham) 21.900 21.300 20.700 22.100 MISSINGa MISSING MISSING 38 1182 4 20.700 19.300 19.300 20.300 MISSINGa MISSING MISSING 39 1183 4 20.500 19.700 18.900 19.900 20 20.300 21.700 40 1184 4 21.800 21 20.500 22 21.600 21.500 22.900 41 1185 4 20.400 19.600 19.600 19.°00 21.200 20.500 21.700 42 11.86 4 20.500 20 20.500 20.400 20.800 20.400 21.900 43 1187 4 21.600 21.100 21.200 MISSiNGc MISSING MISSING MISSING 44 1188 4 20.400 19.500 19.900 21.100 20.200 20.800 21.500 45 1189 4 22.800 21.200 21.600 22.600 22 22.200 23.100 46 1190 4 21.800 20.300 20.700 21.100 21 20.100 21.700 47 1191 4 20.700 19.900 19.800 2u.500 20.600 20.500 21.700 48 1192 4 22.200 20.900 21 22.200 21.600 21.400 22.200 35acrificed after 3 weeks on test. EQOy4Get3 hDied during exposure on Day 12. c Di.ed durinq exposure on Day 19.

Final Report 1-1729.005 TABLE GT4 ANALYSIS OF BODY WEIGHTS 24 HOURS PRIOR TO EXPOSURE REF I`6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 28.000) 27.000) 26.000) 25.000) 24.000) 23.000)* ** ** * 22.000)M**** **** M** ***** 21.000)***** M** ***** M* 20.000)* ** x* **** 19.000) 18.000) 17.000) 16.000) 15.000) 14.000) 13.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 21.583 21.150 21.517 21.275 STD.DEV. .708 1.460 .958 .832 R.E.S.D. .720 1.396 1.007 .971 S. E. M. .204 .421 .277 .240 MAXIMUM 22.900 23.300 23.300 22.800 MINIMUM 20.400 17.900 20.200 20.400 SAMPLE SIZE 12 12 12 12 ALL GROUPS COMBINED **********************R*k***k** ANALYSIS OF VARIANCE TABLE ***************x*************** MEAN 21.381 STD.DEV. 1.012 R.E.S.D. .990 S. E. M. .146 MAXIMUM 23.300 MINIMUM 17.900 SAMPLE SIZE 48 * * SOURCE S M F U 0 S UARES * * BETWEEN GROUPS 1.4873 3 .4958 .47 .7066 x WITHIN GROUPS 46.6858 44 1.0610 * p > 0.05; * TOTAL 48.1731 47 NOT SIGNIFICANT **************-***********************h*************************R*****R*A************A****** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 44 1.53 .2195 ***************h*************************************#***#*********************A**A*M******* * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 24 .48 .7007 * BROWN-FORSYTHE 3, 32 .47 .7072 Q UF MEAN SQUARE E VALUE TAIL PROBABILITY Voo94ser3

Final Report 1-1729.005 TABLE GT5 ANALYSIS OF BODY WEIGHTS ON DAY 7 REF D6'PEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 27.000) 26.000) 25.000) 24.000) 23.000) * 22.000)* * *** 21.000)***** M***** M****** ***** 20.000)M**** *** ** M**** 19.000)* ** 18.000) 17.000) * 16.000) 15.000) 14.000) 13.000) 12.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 20.492 20.642 20.883 20.317 STD.DEV. .667 1.395 .771 .741 R.E.S.D. .604 1.202 .804 .855 S. E. M. .192 .403 .223 .214 MAXIMUM 21.800 22.800 22.200 21.300 MINIMUM 19.200 17.200 19.700 19.300 SAMPLE SI ZE 12 12 12 12 AL L GROUPS COMBINED **********************R******** #*******#********************** ANALYSIS OF VARIANCE TABLE * * SOURCE SUM OF SQUARES * DF MEAN SQUARE F VALUE TAIL PROBABILITY MEAN 20.583 * BETWEEN GROUPS 2.0750 3 .6917 .78 .5099 STD.DEV. .933 * WITHIN GROUPS 38.8717 44 .8834 R.E.S.D. .838 * p > 0.05; S. E. M. .135 * TOTAL 40.9466 47 NOT SIGNII•'ICANT MAXIMUM 22.800 * A * # * * R * R * * * * * * R * * R * * * * R * R # * * * * R * # * * * * * * * * * * * * * * k * * # # R * # # * * * # * * * # # * * * * R * * * # * A * * * * A * * * * * * * * * * MINIMUM 17.200 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 44 1.15 .3394 SAMPLE SI ZE 48 * * R * * * * * * * * # * * * * * * * * * * * * * * * * * # * * * * * * R R * * * * * * * * * * * * * * A * * * * * * * * * * * * * * * # A # A * * * A * * * * * * A * * k * A A * * * * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMEI) TO BE EQUAL * WELCH , 4 .13 3574 * BROWN-FORSYTHE 3, 30 .78 .5129 P.Ua94GBE3

Final Report I-1729.005 TABLE GT6 ANALYSIS OF BODY WEIGHTS ON DAY 14 REF D6TEST D10TEST SIIAM .......................+.......................+.......................+.......................+ VAR 5 EXCLUDED DIED DURING EXPOSURE VALUES MIDPOINTS * TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE 23.200) 22.800) 22.400) 22.000) 21..600) * * * 21.200)** ** * * 20.800) * *** ** *** 20.400)M*** M* M M* 20.000)** ** *** * *** ** 19.600)* 19.200)* * * 18.800) * 18.400) 18.000) 17.600) * 17.200) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 20.450 20.425 STD.DEV. .683 1.139 R.E.S.D. .687 1.031 S. E. M. .197 .329 MAXIMUM 21.600 22.300 MINIMUM 19.300 17.700 SAMPLE SIZE 12 12 20.409 .742 .850 .224 21.700 19.600 11 20.308 .812 .882 .234 21.600 18.900 12 ******************************* ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 20.398 * BETWEEN GROUPS .1390 3 .0463 .06 .9796 STD.DEV. .838 * WITHIN GROUPS 32.1507 43 .7477 R.E.S.D. .834 * p > 0.05; S. E. M. .122 * TOTAL 32.2898 46 NOT SIGNII•'ICANT ***********************************R***************R******************************A********* MAXIMUM 22.300 MINIMUM 17.700 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 43 .53 .6659 **********A*****************#**************************************************************k SAMPLE SIZE 47 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 24 .07 .9749 * OO91 BROWN-FORSYTHE 3, 37 .06 .97 93 ' 1VpC3 ,

Final Report I-1729.005 TABLE GT7 ANALYSIS OF BODY WEIGHTS ON DAY 21 REF D6TEST DIOTEST SHAM .......................+.......................+.......................+.......................+ VAR 6 EXCLUDED DIED DURING EXPOSURE VALUES * * TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE MIDPOINTS 22.800) 22.500) * 22.200)* * ** 21.900)** *** * * 21.600) * 21.300)** * * 21.000)M ** M* M* 20.700)*** M* * 20.400)* * *** *** 20.100)** * * 19.800) ** 19.500) 19.200) * 18.900) 18.600) 1.8.300) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 21.000 20.733 STD.DEV. .659 1.033 R.E.S.D. .720 1.040 S. E. M. .190 .298 MAXIMUM 22.100 21.900 MINIMUM 20.200 18.600 SAMPLE SIZE 12 12 21.045 .674 .721 .203 22.200 20.100 11 21.100 .982 1.075 .296 22.600 19.900 11 *********RR********kR******RR*R ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 20.965 * BETWEEN GROUPS .9304 3 .3101 .42 .7367 STD.DEV. .839 * WITHIN GROUPS 30.7139 42 .7313 R.E.S.D. .859 * p > 0.05; S. E. M. .124 * TOTAL 31.6443 45 NOT SIGNIFICANT ******************************R************************************************************* MAXIMUM 22.600 MINIMUM 18.600 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 42 1.20 .3203 ******R************************************************f******#***************************** SAMPLE SIZE 46 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO RF EQUAL * WELCH 3, 23 .30 .8244 * BROWN-FORSYTHE 3, 36 .42 .7363 1-UOJf./be3

Final Report 1-1729.005 TABLE GT8 ANALYSIS OF BODY WEIGHTS ON DAY 28 REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ VAR 7 EXCLUDED ANIMALS SACRIFICED FOR 3 WEEK ANALYSIS VALUES *** *** *** *** TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE MIDPOINTS 23.400) 23.100) 22.800) 22.500) * 22.200) * 21.900) ** * 21.600)* * ** 21.300) * * 21.000)** * M M 20.700)M* M ** ** 20.400) ** ** *** 20.100) * ** 19.800) * 19.500) * * 19.200) 18.900) *'S, N'S OTIIERWISE MEAN 20.578 20.778 S9'D.DEV. .595 .845 R.E.S.D. .594 .890 S. E. M. .198 .282 MAXIMUM 21.600 22.100 MINIMUM 19.600 19.600 SAMPLE SIZE 9 ALL GROUPS COMBINED MEAN 20.867 STD.DEV. .727 R.E.S.D. .727 S. E. M. .121 MAXIMUM 22.600 MINIMUM 19.600 SAMPLE SIZE 36 R00946eR 21.111 21.000 .777 .671 .847 .709 .2.59 .224 22.6C0 22.000 20.400 20.000 9 9 9 * * * * * * * * * * * M1 * * * * * * * * * * * * * * * * * * * * ANALYSIS OF VARIANCE 'PABLE * * * k * * * * * * * * * * fr * * * * * * * * * * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS 1.5200 3 .5067 .95 .4259 * WITHIN GROUPS 16.9800 32 .5306 * p > 0.05; * TOTAL 18.5000 35 NOT SIGNIFICANT ***************************************k*************l.********************i.***************** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 32 .60 .6206 *****************************k**********#**********k**************************************** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 18 * BROWN-FORSYTHE 3, 30 1.05 .3929 .95 .4267

Final Report 1-1729.005 TABLE GT9 ANALYSIS OF BODY WEIGHTS ON DAY 35 REF D6TEST DIOTEST SHAM ............+ .......................+.......................+.......................+ VAR 8 EXCLUDED ANIMALS SACRIFICED FOR 3 WEEK ANALYSIS VALUES *** *** h** k** MIDPOINTS TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE 23.400) 23.100) 22.800) * 22.500) * 22.200) * 21.900) * * 21.600)*** * 21.300)* M* * 21.000)N * ** N 20.700)** N *** * 20.400)*** ** **** 20.100) *k* * 19.800) 19.500) * 19.200) 18.900) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 20.933 20.678 STD.DEV. .539 .894 R.E.S.D. .640 .935 21.267 .684 .640 20.856 .695 .748 S. E. M. .180 .298 MAXIMUM 21.600 22.400 MINIMUM 20.300 19.600 SAMPLE SIZE' 9 9 .228 22.800 20.600 9 .232 22.200 20.100 9 ALL GROUPS COMBINED ***********************R*k***** ******************************* ANALYSIS OF VARIANCE TABLE * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 20.933 * BETWEEN GROUPS 1.6422 3 .5474 1.07 .3742 STD.DEV. .716 * WITHIN GROUPS 16.3178 32 .5099 R.E.S.D. .709 * p > 0.05; S. E. M. .119 * TOTAL 17.9600 35 NOT SIGNIFICANT MAXIMUM 22.800 ***********************************k****************************R*************************** MINIMUM 19.600 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 32 .65 .5861 SAMPLE SIZE 36 ***k*************************************************************************************k** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO HE EQUAL * WELCH 3, 18 .91 .4565 * E;00946gE3 BROWN-FORSYTHE 3, 28 1.07 .3762

Final Report 1-1729.005 TABLE GT10 ANALYSIS OF BODY WEIGHTS ON DAY 42 VAR 9 EXCLUDED VALUES REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ ANIMALS SACRIFICED FOR 3 WEEK ANALYSIS * * * TABULATIONS AND MIDPOINTS *** *** COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE R## 24.300) 24.000) 23.700) 23.400) * 23.100) * * 22.800)* * 22.500)** 22.200)M* * 21.900)* * ** M 21.600)* M* N ***** 21.300)** **# * 21.000) M* 20.700) * 20.400) * * 20.100) * 19.800) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 22.089 21.500 STD.DEV. .540 1.099 R.E.S.D. .578 .857 S. E. M. .180 .366 MAXIMUM 22.900 24.000 MINIMUM 21.300 20.000 SAMPLE SIZE 9 9 21.567 1.005 1.073 .335 23.300 20.500 9 22.044 .577 .611 .192 23.100 21.500 9 **#*R*************R***R******** ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 21.800 * BETWEEN GROUPS 2.5889 3 .8630 1.21 .3202 STD.DEV. .851 * WITHIN GROUPS 22.7311 32 .7103 R.E.S.D. .745 * p > 0.05; S. L•'. M. .142 * TOTAL 25.3200 35 NOT SIGNIFICANT ***************#****#***********************#*****R#****************R***#*****#************* MAXIMUM 24.000 MINIMUM 20.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 32 .92 .4417 **#**********#*******************i.******k****#**R*****R************************k*#********** SAMPLE SIZE 36 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3 17 1.13 .3646 6L4946grj * BROWN-FORSYTHE , 3, 24 1.21 .3258

Final Report I-1729.005 Table GT11 Weight of Lung Tissue at Necropsy in Mice Exposed to Cigarette Smoke for 6 Weeks Group-Cigarette Animal Number Lung Weight (g) 1-Reference 1151 1152 1153 0.13 0.15 0.15 Mean + Stand. Dev. 0.14 + 0.01 2-D6 1163 0.13 1164 0.14 1165 0.17 Mean + Stand. Dev. 0.15 + 0.02 3-D10 1175 0.17 1176 0.15 1177 0.15 Mean + Stand. Dev. 0.16 + 0.01 4-Sham 1186 0.14 1188 0.12 1189 0.16 Mean + Stand. Dev. 0.14 + 0.02

Final Report I-1729.005 Table GT12 Aryl Hydrocarbon Hydroxylase (AHH) Activity in Selected Tissues of Mice Exposed to Cigarette Smoke for Six Weeksa b Tissue AHH Values + SD Group Liver Lung Kidney (pmol/min/mg protein) ----------------------------------------------------=----------------- 1-Ref 109.9 + 61.5 2.1 + 0.6 0.7 + 0.3 2-D6 80.0 + 62.6 0.9 + 0.8 1.0 + 0.3 3-D10 76.5 + 46.6 1.7 + 0.8 0.9 + 0.2 4-Sham 42.6 + 17.8 2.1 + 1.7 0.5 + 0.2 aMice were exposed daily (5 days/week) using the RSD1 exposure regimen for 6 weeks. Animals were sacrificed 6 hours after initiation of the last exposure to determine tissue levels of AHH. bAHH units are pmoles 3-OH-BaP formed/min/mg microsomal protein at 37oC for 20 minutes. Means + standard deviations are given.

Final Report 1-1729.005 Abbreviations for Clinical Chemistry and Hematology BUN - Blood urea nitrogen CALC - Calcium CREAT - Creatinine LDH - Lactic dehydrogenase ALKPHOS - Alkaline phosphatase P04,PHOS - Phosphorous SGOT - Serum-glutamic-oxaloacetic-transaminase SGPT - Serum-glutamic-pyruvic-transaminase TGA - Triglycerides URICACID - Uric acid AGRATIO - Albumin/globulin ratio WBC - White blood cell RBC - Red blood cell HGB - Hemoglobin HCT - Hematocrit NEUTS - Segmented neutrophils LYMPHS - Lymphocytes MONO - Monocytes RETCTN - Reticulocyte counts CHOL - Cholesterol TOTPROT - Total Protein PMNCTN - Polymorphonuclear leukocyte counts

Final Report I-1729.005 Abbreviations of BMDP Statistical Tables STD. DEV. - Standard Deviation R.E.S.D. - Robust Estimate of Standard Deviation S.E.M. - Standard Error of the Mean N.S. - Not Significant, p > 0.05

Final Report 1-1729.005 TABLE CCH1 CLINICAL CHEMISTRY FOR 3 WEEK SACRIFICE OF MICE NO. LABEL CIG GLUCOSE CHOL TGA CREAT BUN URIC ACID TOT PROT ALBUMIN A/G RATIO TYPE mg/dl mg/dl mg/dl mg/dl mg/dl mg/dl g/dl g/dl ---- -------- ---------- ---------- ---------- ---------- ---------- --------- --------- ---------- --- - ------ ---------- 1 1145 REF 121.350 79.500 77.700 .320 30.500 4.050 4.700 2.900 1.611 2 1146 REF 188.350 80.300 69.900 .310 34 4.650 5 2.900 1.381 3 1147 REF 137.300 91.100 66.700 .350 30.750 3.950 4.900 3 1.579 4 1157 D6 156.200 69.900 81.100 .380 30.250 3.350 4.700 2.700 1.350 5 1158 D6 139.200 86.300 87.200 .390 28.450 2.450 4.900 2.900 1.450 6 1159 06 158.600 87 73.300 .440 32.900 4.300 5 3 1.500 7 1169 D10 137.200 87.400 52.500 .400 31.750 4.050 5.200 3 1.364 8 1170 D10 135.350 81.400 55.700 .320 31.350 4.200 4.700 2.900 1.611 9 1181 SHAM 168.400 92.600 58.400 .170 20.500 3.850 5.100 3 1.429 10 1182 SHAM 152.100 84.700 47.400 .110 22.200 4.300 5 3.100 1.632 S FQ9&(;&.7

Final Report 1-1729.005 TA13LE CC112 CL.INICAL CIIh;MIS'PRY FON 3 WEEK SACRI[•'ICN Oh' MICE NO. LABEL LU1I U/1 ALKPIIOS U/1 SGP'1' U/1 SGOT U/1 CAL.:IUM my/c11 PHOS my/dl ---- 1 -------- 1145 ---------- 394 ---------- 71 ---------- 45 ---------- 122 ---------- 10.000 ---------- 6.700 2 1146 300 66 38 213 9.896 7.300 3 1147 215 62 32 124 9.063 7.900 4 1157 183 71 48 147 9.583 6.400 5 1158 201 65 40 QNS`' 10.208 7.100 6 1159 281 78 51 137 10.000 7.200 7 1.169 264 64 39 161 9.583 7.800 8 1170 246 60 29 QNS 9.479 7.300 9 1181 121 66 35 92 9.375 6.900 10 1182 222 50 45 QNS 8.040 6 `'QNS = QIJANTITY NCYI' SUl'Ftc;[RN'I' i•'clk ANALYSTS. ytoy4sea

Final Report I-1729.005 TABLE CCti3 IIEMATOLOGY FOR 3 WEEK SACRIFICE OF MICE NO. LABEL CIG '1'YPE WBC thousand RBC IIGB million y% I1C'I' NEUTS LYMPIIS MONOCYTE $ $ $ $ RETCTN $ PLATELET ---- -------- -- -------- ---------- ---------- ---------- 1 1145 REF 5.400 9.570 15.800 47.600 31 68 1 .800 ADEQ 2 1146 RL•'F 5.400 10.790 17.800 52.300 41 57 1 .600 ADEQ 3 1147 REF 3.700 9.090 15.100 43.900 32 67 1 1.400 ADEQ 4 1157 D6 4.900 9.920 16 47.900 26 70 2 0 1.500 ADEQ 5 1158 D6 4 9.100 14.700 44.600 28 72 NR 1.500 ADEQ 6 1159 D6 5.600 10.130 16.600 49.400 23 74 NR .800 ADEQ 7 1169 D10 4.300 9.270 14.600 44.900 33 64 1 1.100 A[)EQ 8 1170 D10 6.100 9.660 16.500 47.900 21 76 NR .900 ADEQ 9 1181 SNAM 1.600 8.870 14.700 43.300 38 62 NR 1.600 ADEQ 10 1182 SIiAM 2.300 9.760 15.500 47.500 38 62 NR 1.500 ADEQ u N 12 = NONE RL:L'OR'I'C:D OR OBSI•IIiVP7D. 4toyj,sae

Final Report 1-1729.005 TABLE CCH4 HEMATOLOGY FOR 3 WEEK SACRIFICE OF MICE NO. LABEL PMNCTN EOSIN $ ------ 1 2 1145 1146 MODERATE MODERATE a NR 1.0 3 1147 MODERATE NR a 4 1157 MODERATE 2.0 5 1158 MODERATE NRa 6 1159 MODERATE 3.0 7 1169 MODERATE 2.0 8 1170 MODERATE 3.0 9 1181 MODERATE NR a 10 1182 MODERATE NRa a NOT REPORTED OR OBSERVED. Nio94sea

Final Report 1-1729.005 TABLE CCH5 HISTOGRAM OF GLUCOSE ANALYSIS OF GLUCOSE VALUES AFTER ; WEEKS EXPOSURE REF D6TEST D10TEST SHAM ......................... ....................................................................... .....+ MIDPOINTS 200.000) 190.000) * 180.000) 170.000) 160.000) ** 150.000)N N 140.000)* * M* 130.000) 120.000) * 110.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 149.000 STD.DEV. 34.999 R.E.S.D. 40.268 S. E. M. 20.207 MAXIMUM 188.350 MINIMUM 121.350 SAMPLE SIZE 3 ALL GROUPS COMBINED MEAN 149.405 STD.DEV. 19.436 R.E.S.D. 16.001 S. E. M. 6.146 MAXIMUM 188.350 MINIMUM 121.350 SAMPLE SIZE 10 151.333 136.275 10.576 1.308 12.416 1.640 6.106 .925 158.600 137.200 139.200 135.350 3 2 k**********k******R***RkR****** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH ' * BROWN-FORSYTHE * N * 160.250 11.526 14.445 8.150 168.400 152.100 2 ANALYSIS OF VARIANCE TABLE *******R*****k***************** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS 591.6689 3 197.2230 .42 .7446 * WITHIN GROUPS 2808.0981 6 468.0163 * p > 0.05; * TOTAL 3399.7666 9 NOT SIGNIFICANT ***********************************************Rk********************R******k*****k*****R*** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 4.50 .0559 **RR**********************************R****************************************R************ 3, 3 3.20 .1823 3, 3 .57 .6736 G10y46B3

Final Report 1-1729.005 TABLE CCH6 HISTOGRAM OF cHOL ANALYSIS OF CHOLESTEROL AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 96.000) 93.000) * 90.000)* N 87.000) 84.000)N * * * N * 81.000)* N 78.000) * 75.000) 72.000) 69.000) * * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 83.633 81.067 84.400 88.650 STD.DEV. 6.479 9.677 4.243 5.586 R.E.S.D. 7.641 11.427 5.317 7.001 S. E. M. 3.740 5.587 3.000 3.950 MAXIMUM 91.100 87.000 87.400 92.600 MINIMUM 79.500 69.900 81.400 84.700 SAMPLE SIZE 3 3 2 2 * i. * * * *R****R#*********************** ANALYSIS OF VARIANCE TABLE ALL GROUPS COMBINED * * * * * * R * * k * * * * * * * * A k * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY MEAN 84.020 * BETWEEN GROUPS 69.7777 3 23.2592 .44 .7356 STD.DEV. 6.585 * WITHIN GROUPS 320.4386 6 53.4064 R.E.S.D. 6.760 * p > 0.05; S. E. M. 2.082 * TOTAL 390.2163 9 NOT SIGNIFICANT *****#*****R******************R*************k*******R***********************************k*** MAXIMUM 92.600 MINIMUM 69.900 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.83 .2426 ***************R*******************A*******************R**#*************************k******* SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 .35 .7932 * BROWN-FORSYTHE 3, 5 .52 .6871 ozos4sea

TABLE CCH7 ANALYSIS OF TRIGLYCERIDES AFTER 3 WEEKS EXPOSURE IIIS'I'OGRAM OL•' TGA Final Report 1-1729.005 REF D6TEST D10'I'h:ST S[IAM .......................4 ....................... i....................... -I................ ........t M1DPOIN'/'S 90.000) 85.000) 80.000)* 75.000) 70.000) M 65.000)* 60.000) * M * * 55.000) M N 50.000) 45.000) * dGROUP MEANS ARE DENO'PEI) BY M' S IF THEY COINCIDE WITII *'S , N'S OTIIERWISE MEAN 71 433' a,c 8 1), ° . 0.533 54.100 52.900 STI"1.Ij EV. 5.658 6.967 2.263 7.778 R,E.S.D. 6.413 7.402 2.836 9.748 S. E. M. 3.267 4.023 1.600 5.500 MAXIMUM 77.700 87.200 55.700 58.400 MINIMUM 66.700 73.300 52.500 47.400 SAMPLE SIZE 3 3 2 2 ALL GROUPS COMI3INE1) *****k*#****k*#**#k*#***k*kk*** ANALYSIS OE' VARIANCE 'I'AU[,E *******k***#*k*#k*Akkk**k****#k k * SOURCE SUM OF SQUARES DE MEAN SOUARE F VALUE TAIL PROBABILITY * MEAN 66.990 * BEITWEEN GROUPS 1338.8557 3 446.2852 11.81 .0063 STD.DEV. E 13.189 * * 4'ITIIIN GROUPS 226.7333 6 37.7889 R. .S.U. 5.443 S. E. M. 4.171 * TOTAL 1565.5889 9 05 1) ` 0 MAXIMUM 87.200 k * # # # * * * k # # * * k * k * * * * * # k * * # k * * k * k * * * k k k * * * * # * # * * # * * * k * * * * k # * A * * * * # # # * * * # * * k * . * * * k k * * k # * * * # * # * fi MINIMUM 47.400 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 .92 .4846 SAMPLE SI'LE 10 k # * # # * * # * * # * * * * * k * * * # * * * # k # * * * * k * * * * * # k * * * * * * k * * * * * k * * A * k * F * i. * * # * # * * * * * * * * * A k * * * * k * * k k # * * * * * * ONE-WAY ANALYSIS OE' VARIANCE * TEST S'I'ATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EpUAL * WELCH 3, 3 12.17 .0347 * BRUWN-E'ORSY'1'[iE 3, 4 12.30 .0173 tzos4sse aSIGNIFICAN'1'LY D1FE'13R1•'N'I' PROM 5[IAM CON9'ROL, S'PIIDI:N'I'1'LGIi RANG!: '['I:ST, p< 0.()5 bSIGNIE'ICANTLY DIFFLRPIN'C FROM REFERENCE C1GAR1;'Nl'I:, i'!'[1DiN'1'l'l.E[) RANGE TIi 9', p 0,05, cS1GN[I•'ICANTLY DI1'NI:REN'P FROM OTHER TEST CIG,IRE1'TL•', S'I'UI)I;N'I'I'LI,D RANGI•1 TEST, p`< 0,05.

Final Report I-1729.005 tl I S'l'OGRAM 01' CItEIA'1' TABLE CCH8 ANALYSIS OF CREATININE AFTER 3 WEEKS EXPOSURE kEF 1)6'I'ES'I' D10'I'I:S'1' SIIAM ............................................... i.............................. .................. MIDPOIN'PS .500) .450) .400) .350) M .300)** .250) .200) .150) .100) .050) "GROUP MEANS ARE DIiNO`I'ED BY M'S IF * M* * N * MEAN .32V' S'1'D.DEV. .021 R.E.S.D. .024 S. E. M. .012 MAXIMUM .350 MINIMUM .310 SAMPI,E SIZE 3 AL,L GROIIPS COMBINED MEAN .319 STD.DEV. .104 R.E.S.D. .034 S. E. M. .033 MAXIMUM .440 MINIMUM .110 SAMPLE SIZE 10 M * 'f'HEY COINCIDE WITtI *'S, N'S OTIIE]RWIShI 403 ~ . 360 a .032 .057 .038 .071 .019 .040 .440 .400 .380 .320 3 2 * * * k * A * * * * * * * * * * * * * * * * * * * * k * * * * ANALYSIS * * * /. * * * * * A * * * * * * * * k * * * * * * * * * * k * * * SOURCE SUM OF SQUARES DE MEAN S~,)IIARE F VALUE TAII, PROBABILITY * * BETWEEN GROUPS .0890 3 .0297 22.43 .0012 * WITHIN GROUPS .0079 6 .0013 * * TOTAL .0969 9 1) < 0.05 * A * * * * * * * * * * * * * * A * * * * A * * * A * * * * * * * * k * * * * * * A * l. * * * * * * * * * * * * * * * * * * * * * * A * * * * * * * * * A * * R A * * * * * * * * * A * * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 3.66 .0828 * * * * * * * * * * * k * * * * * A * * * * * * * * * * * * * A * * * * * * k * * * * * * k * * * * * * * * A * * * A R A k * * * * * * * * * * * k * A A A A A * * * * * * * * * * A * * ONE-WAY ANALYSIS OF VARIANCE * TEST STA'I'ISTICS FOR WITIIIN-GROI)P * VARIANCES NO'I' ASSUMED '1'0 BE EQUAL * WELCII 3, 2 * BROWN-PORSY't'lIE 3, 3 12.10 .0773 17.70 .0207 a 5IGN1l'lt'AN'I'I,Y UL1•'NI:RI-N'I' f'lt(1M SunM CONTRC)I,, tPl'lID1:N'1'IZP,I) kAN(J; < 0.05. zzo94seR

Final Report 1-1729.005 HISTOGRAM ON 6UN 12E;E' TABLE CCH9 ANALYSIS OF BLOOD UREA NITROGEN AFTER 3 WEEKS EXPOSURE D 6'1' E S'1' D 1. 0'I' E S'I' S 11 A M ....................... +............................................... MIDPOIN'I'S t..................... ..+ 39.000) 36 000) . 33.000)* 30.000)N 27.000)** # M* N 24.000) 21.000) 18.000) 15.000) 12.000) 'GROUP MEANS ARE DENOTED BY 'S * F * * HEY COINCIDE WITU *'S, N'S OTHERWISE * M * MEAN 29.867a 26.367a 25.567 a 17.667 STD.DEV. 3.848 1.856 3.580 2.281 R.E.S.D. 4.537 1.978 4.025 2.524 S. E. M. 2.221 1.071 2.067 1.317 MAXIMUM 34.300 28.300 29.500 19.700 MINIMUM 27.400 24.600 22.500 15.200 SAMPLE SIZE 3 3 3 3 ALL GROUPS COMBINED *************************R****# ANALYSIS Oh * * * # * k * * * * * * * * * * * * * k * * * k k * A * * * * VARIANCE TABLE x * SOURCE SUM OE' SQUARES 1)E MEAN SQUARE F VALUE 'fAIL k PROBABILITY MEAN 24.867 * BETWEEN GROUPS 238.7401 3 79.5800 8.78 .0065 STD.DEV. 5.319 * WITHIN GROUPS 72.5267 8 9.0658 R.E.S.D. 2.785 * S. E. M. 1.536 * TOTAL 311.2667 11 p< 0.05 MAXIMUM 34.300 * * * * R * * * * * * * * # * * * * * * * * * k * * * * * * * * * * M * * * * k * * * * * k # * * * * * * A * * * * k * * * A k * * * A * * # * * * * * * * * * * * * * * * * * * * * * MINIMUM 15.200 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 1.19 .3746 SAMPLE S1 ZE 12 * * * * R R * * * * * * * * * * * * * * * * * * k * # * * * * * * * * * * # * * * * * * * A * * * * * * R k * k * * * * * * k * * * M # * # # * # k * * k * * * * * * * k * * # * * k # * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITUIN-GROUP * VARIANCES NO'I' ASSUMED TO BE EQUAL * WELCII 3, 4 9.04 0296 * BROWN-E'ORSY'PUE 3, 6 8.78 .0130 dSIC;NIFICAfITLY DIFI•'ERI:N'P I'ROM SUAM CONTROL, S'TUDI:N'I'I'LL•'f) kAN(;1: 'I'RS'I', 1< 0.05. EZ03G6e9

Final Report I-1729.005 TABLE CCH10 HISTOGRAM OF URICACID ANALYSIS OF URIC-ACID AFTER 3 WEEKS EXPOSURE REF D6TEST DIOTEST SHAM .................+.......................+.......................+.......................+ MIDPOINTS 4.800) 4.500) * 4.200)N * M M 3.900)** * * 3.600) 3.300) M 3.000) 2.700) 2.400) * 2.100) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 4.217 3.367 STD.DEV. .379 .925 R.E.S.D. .443 .955 S. E. M. .219 .534 MAXIMUM 4.650 4.300 MINIMUM 3.950 2.450 SAMPLE SIZE 3 3 4.125 .106 .133 .075 4.200 4.050 2 4.075 .318 .399 .225 4.300 3.850 2 ****R*t*****A*********R******** ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE * * k * * * * * M * * * * * * * * A A * A * * * * * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 3.915 * BETWEEN GROUPS 1.3144 3 .4381 1.25 .3733 STD.DEV. .617 * WITHIN GROUPS 2.1108 6 .3518 R.E.S.D. .440 * p > 0.05; S. E. M. .195 * TOTAL 3.4253 9 NOT SIGNIFICANT *****R*****i.f***************i**********A**********************************A***********M****k MAXIMUM 4.650 MINIMUM 2.450 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.41 .3289 ***************************************************************************A*****x********** SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 .50 .7056 * BROWN-FORSYTHE 3, 3 1.67 .3428 Vzousetq

Final Report I-1729.005 TABLE CCH11 HISTOGRAM OF TOTPROT ANALYSIS OF TOTAL PROTEIN AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 5.250) 5.180) 5.110) 5.040) * 4.970)* * N 4.900)M M 4.830) 4.760) 4.690)* 4.620) * * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 4.867 STD.DEV. .153 R.E.S.D. .171 S. E. M. .088 MAXIMUM 5.000 MINIMUM 4.700 SAMPLE SIZE 3 ALL GROUPS COMBINED MEAN 4.920 STD.DEV. .175 R.E.S.D. .167 S. E. M. .055 MAXIMUM 5.200 MINIMUM 4.700 SAMPLE SIZE 10 4.867 4.950 .153 .354 .171 .443 .088 .250 5.000 5.200 4.700 4.700 3 2 ********R********************** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH * BROWN-FORSYTHE * N * 5.050 .071 .089 .050 5.100 5.000 2 ANALYSIS OF VARIANCE TABLE * * * * * * * * l. * * * * * * * * * * * * * * * * * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS .0527 3 .0176 .47 .7131 * WITHIN GROUPS .2233 6 .0372 * p > 0.05; * TOTAL .2760 9 NOT SIGNIFICANT *********!R********************Ie******************************it*******k*********1c*********** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 4.53 .0551 ******************************************************************************************** 3, 3 1.19 .4439 3, 2 .39 .7783 ~zos,~se~

Final Report I-1729.005 TABLE CCH12 IIISTOGRAM OF' ALBUMIN ANALYSIS OF ALBUMIN AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................................................... ................+ MIDPOINTS 3.220) 3.150) 3.080) M 3.010) * * * * 2.940) N N 2.870)** M * 2.800) 2.730) * 2.660) 2.590) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 2.933 2.867 STD.DEV. .058 .153 R.E.S.D. .068 .171 S. E. M. .033 .088 MAXIMUM 3.000 3.000 MINIMUM 2.900 2.700 SAMPLE SIZE 3 3 2.950 .071 .089 .050 3.000 2.900 2 3.050 .071 .089 .050 3.100 3.000 2 ALL GROUPS COMBINED ****R*k***#***R*k**********#**R ***************R*************** ANALYSIS OF VARIANCE TABLE * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY MEAN R 2.940 * BETWEEN GROUPS .0407 3 .0136 1.28 .3621 STD.DEV. R.E.S.D. .107 * .088 * WITHIN GROUPS .0633 6 .0106 p> 0.05; S. E. M. .034 * TOTAL .1040 9 NOT SIGNIFICANT MAXIMUM 3.100 k*R*******#*******************************************************************************hR MINIMUM 2.700 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.64 .2766 SAMPLE SIZE 10 *#R*#**k*********#*R**##*#*************#****#!*R***********#***********************k****#*** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 1.14 .4592 * BROWN-FORSYTHE 3, 4 1.52 .3376 szo94set3

Final Report 1-1729.005 TABLE CCH13 HISTOGRAM OF AGRATIO ANALYSIS OF A/G RATIO AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 1.680) 1.640) * 1.600) * * 1.560) * 1.520)N N 1.480) * N 1.440) M  1.400)* 1.360) 1.320) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S , N'S OTHERWISE MEAN 1.524 1.433 STD.DEV. .125 .076 R.E.S.D. .146 .085 S. E. M. .072 .044 MAXIMUM 1.611 1.500 MINIMUM 1.381 1.350 SAMPLE SIZE 3 3 1.487 .175 .219 .124 1.611 1.364 2 1.530 .144 .180 .102 1.632 1.429 2 AL[. GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 1.491 * BETWEEN GROUPS .0163 3 .0054 .35 .7937 STD.DEV. .111 * WITHIN GROUPS .0940 6 .0157 R.E.S.D. .119 * p > 0.05; S. E. M. .035 * TOTAL .1102 9 NO'I' SIGNIFICANT MAXIMUM 1.632 ***Rk****~F********************~F************#**h***R*k*fr**ir*8.F*******i4***********RM*kR******* MINIMUM 1.350 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.96 .2221 SAMPLE SIZE 10 *****1F*************************************fi***************it***********************1x******** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 2 .35 .7985 * BROWN-FORSYTHE 3, 3 .29 .8314 LzOyL6B~q

Final Report I-1729.005 TABLE CCH14 HISTOGRAM OF LDH ANALYSIS OF LACTIC DEHYDROGENASE AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 440.000) 400.000)* 360.000) 320.000)N 280.000)* * * 240.000) N M * 200.000)* ** 160.000) N 120.000) * 80.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S , N'S OTHERWISE MEAN 303.000 221.667 STD.DEV. 89.538 52.166 R.E.S.D. 93.123 60.717 S. E. M. 51.695 30.118 MAXIMUM 394.000 281.000 MINIMUM 215.000 183.000 SAMPLE SIZE 3 3 255.000 12.728 15.952 9.000 264.000 246.000 2 171.500 71.418 89.509 50.500 222.000 121.000 2 ALL GROUPS COMBINED ******************************* ******************************* ANALYSIS OF VARIANCE TABLE * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 242.700 * BETWEEN GROUPS 22676.9336 3 7558.9775 1.70 .2662 STD.DEV. 74.099 * WITHIN GROUPS 26739.1667 6 4456.5278 R.E.S.D. 55.443 * p > 0.051 S. E. M. 23.432 * TOTAL 49416.0938 9 NOT S1GN1F1CANT MAXIMUM 394.000 ******************************************************************************k**k********** MINIMUM 121.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.18 .3933 SAMPLE SIZE 10 ****k******************************************************************i.*************fi****** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 1.03 .4899 * BROWN-FORSYTHE 3, 4 1.93 .2659 Rzas4sea

Final Report 1-1729.005 TABLE CCH15 HISTOGRAM OF ALKPHOS ANALYSIS OF ALKALINE PHOSPHATASE AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 80.000) 76.000) 72.000)* 68.000)N 64.000)* 60.000)* 56.000) 52.000) 48.000) 44.000) GROUP MEANS ARE DENOTED BY M'S * M * * M * N * IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 66.333 71.333 62.000 58.000 STD.DEV. 4.509 6.506 2.828 11.314 R.E.S.D. 4.776 6.822 3.545 14.180 S. E. M. 2.603 3.756 2.000 8.000 MAXIMUM 71.000 78.000 64.000 66.000 MINIMUM 62.000 65.000 60.000 50.000 SAMPLE SIZE 3 3 2 2 ALL GROUPS COMBINED *k**********A****************** ANALYSIS * * OF VARIANCE TABLE * * k * * * * * k A * * * * * k * * * * * * * k k * * * * A * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE 'PAIL PROBABILITY * MEAN 65.300 * BETWEEN GROUPS 240.7667 3 80.2556 1.84 .2400 STD.DEV. 7.469 * WITHIN GROUPS 261.3333 6 43.5556 R.E.S.D. 5.637 * p > 0.05; S. E. M. 2.362 * TOTAL 502.1000 9 NOT SIGNIFICANT ********A*#***************************A********************************k******************** MAXIMUM 78.000 MINIMUM 50.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 2.28 .1793 ********************k******A***********A****************************M******AR*k************* SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 1.36 .4026 * BROWN-FORSYTHE 3, 2 1.58 .4107 f,zoy4sea

Final Report 1-1729.005 TABLE CCH16 HISTOGRAM OF SGPT ANALYSIS OF SGPT AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 54.000) 51.000) * 48.000) * 45.000)* N * 42.000) 39.000)M * N 36.000) * 33.000) * N 30.000) * 27.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 38.333 46.333 STD.DEV. 6.506 5.686 R.E.S.D. 6.822 6.481 S. E. M. 3.756 3.283 MAXIMUM 45.000 51.000 MINIMUM 32.000 40.000 SAMPLE SIZE 3 3 34.000 7.071 8.862 5.000 39.000 29.000 2 40.000 7.071 8.862 5.000 45.000 35.000 2 ALL GROUPS COMBINED ******************************* ANALYSIS * * * * * OF VARIANCE TABLE * * * * * * * * * * * * * * * * * * * * * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL. PROBABILITY * MEAN 40.200 * BETWEEN GROUPS 200.2667 3 66.7556 1.61 .2840 STD.DEV. 7.068 * WITHIN GROUPS 249.3333 6 41.5556 R.E.S.D. 6.077 * P > 0.05; S. E. M. 2.235 * TOTAL 449.6000 9 NOT SIGNIFICANT *******************************k******************************************************w***** MAXIMUM 51.000 MINIMUM 29.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 .06 .9778 *********************A********************************A************************************* SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 1.15 .4557 * BROWN-FORSYTHE 3, 4 1.51 .3399 UE09L6gR

Final Report 1-1729.005 TABLE CCH17 ANALYSIS OF SGOT AFTER 3 WEEKS EXPOSURE HISTOGRAM OF SGOT REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ VAR 16 EXCLUDED VALUES QUANTITY NOT SUFFICIENT FOR ANALYSIS * * * MIDPOINTS 240.000) TABULATIONS AND COMPU;ATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE 220 0 * 0 . ) 0 200.000) 180.000) 160.000)N M 140.000) M* 120.000)** 100.000) M 80.000) 60.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 153.000 142.000 STD.DEV. 51.971 7.071 R.E.S.D. 61.400 8.862 S. E. M. 30.006 5.000 MAXIMUM 213.000 147.000 MINIMUM 122.000 137.000 SAMPLE SIZE 3 2 161.000 .000 .000 .000 161.000 161.000 1 92.000 .000 .000 .000 92.000 92.000 1 ********k********************** ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE R****************************** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 142.286 * BETWEEN GROUPS 3223.4287 3 1074.4763 .59 .6617 STD.DEV. 38.025 * WITHIN GROUPS 5452.0000 3 1817.3333 R.E.S.D. 25.141 * p > 0.05; S. E. M. 14.372 * TOTAL 8675.4277 6 NOT SIGNIFICANT *************************k******M****k**********************k*****k************************* MAXIMUM 213.000 MINIMUM 92.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 3 4.05 .1405 **************k*********************k*****R*****A******k***A****k****k********R************* SAMPLE SIZE 7 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 11 .04 .9893 * BROWN-FORSYTHE 3, 2 2.04 .3455 I(:Q9468R

Final Report I-1729.005 TABLE CCH18 HISTOGRAM OF CALCIUM ANALYSIS OF CALCIUM AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 10.400) 10.200) * 10.000)* M 9.800) * 9.600) N * M 9.400) * * 9.200) 9.000) * N 8.800) 8.600) * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 9.653 9.931 STD.DEV. .514 .318 R.E.S.D. .604 .355 S. E. M. .297 .184 MAXIMUM 10.000 10.208 MINIMUM 9.063 9.583 SAMPLE SIZE 3 3 9.531 .074 .092 .052 9.583 9.479 2 9.010 .516 .646 .365 9.375 8.646 2 ***R*#************************* ALL GROUPS COMBINED ANALYSIS * * * # OF VARIANCE TABLE * * k w * * * * * * * * * * * * * k * k k * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 9.583 * BETWEEN GROUPS 1.0381 3 .3460 2.07 .2054 STD.DEV. .476 * WITHIN GROUPS 1.0019 6 .1670 R.E.S.D. .358 * p > 0.05 S. E. M. .151 * TOTAL 2.0399 9 NOT SIGNIFICANT ***********************************~F*************************************R**#R************** MAXIMUM 10.208 MINIMUM 8.646 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 3.10 .1108 ***************************************************#************************.*************** SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 1.49 .3750 * BROWN-FORSYTHE 3, 3 2.20 .2675 v(:Q9z1b&R

Final Report I-1729.005 TABLE CCH19 HISTOGRAM OF PHOS ANALYSIS OF PHOSPHORUS AFTER 3 WEEKS EXPOSURE REF D6TEST D10TEST SHAM ....................... +........................................................................ .......+ MIDPOINTS 8.400) 8.100) 7.800)* * 7.500) N 7.200)M ** * 6.900) N * 6.600)* 6.300) * N 6.000) * 5.700) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S , N'S OTHERWISE MEAN 7.300 6.900 7.550 6.450 STD.DEV. .600 .436 .354 .636 R.E.S.D. .614 .512 .443 .798 S. E. M. .346 .252 .250 .450 MAXIMUM 7.900 7.200 7.800 6.900 MINIMUM 6.700 6.400 7.300 6.000 SAMPLE SIZE 3 3 2 2 ALL GROUPS COMBINED #*#R*R*R*****R*R*#R******R**RRR ANALYSIS OF VARIANCE TABLE ******************************R * " SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 7.060 * BETWEEN GROUPS 1.4740 3 .4913 1.81 .2458 STD.DEV. .587 * WITHIN GROUPS 1.6300 6 .2717 R.E.S.D. .476 * p > 0.05; S. E. M. .186 * TOTAL 3.1040 9 NOT SIGNIFICANT MAXIMUM 7.900 ******R*****A************#*R***************#**RR****A****#**********R***k******************* MINIMUM 6.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 .33 .8043 SAMPLE SIZE 10 *****R*R**R*R**R*R**R*R*******RR*R****R*#k**#*****##*********R****R*********k*#R****#******* * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 1.40 .3949 * BROWN-FORSYTHE 3, 4 1.82 .2831 f3:Q9F.t6g d

Final Report 1-1729.005 TABLE CCH2O ANALYSIS OF WHITE BLOOD CELL COUNTS AFTER 3 WEEKS EXPOSURE IIIS'L'OGRAM OF WBC RP:L' D6'PES'I' UlOTEST SIIAM .......... ..................................... /....................... i........................ 1- MI1)POINTS 9.000) 8.000) 7.000) 6.000) 5.000) M* 4.000) * 3.000) 2.000) 1.000) .000) aGRUUP MEANS E DENOTED Y S * M * L, IEY COINCIDE WITH * N * 'S, S IIERWISE * MEAN 4.833 a 4.833a 5.200° 1.950 STD.DEV. .981 .802 1.273 .495 R.E.S.D. 1.160 .853 1.595 .620 S. E. M. .567 .463 .900 .350 MAXIMUM 5.400 5.600 6.100 2.300 MINIMUM 3.700 4.000 4.300 1.600 SAMPLE SIZE 3 3 2 2 * * * ALL GROUPS COMBINED * * * * * * k * * * * * * * * * A * * * * * * * * * * * * * * * * * * * * * * * * * * ANALYSIS OF VARIANCE TABLE * * * * * * * * * * * * * * * * * * SOURCE SUM OF SQUARES DP MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 4.330 * BETWEEN GROUPS 14.3627 3 4.7876 5.66 .0349 STD.DEV. 1.470 * WITIIIN GROUPS 5.0783 6 .8464 R.E.S.D. .850 * S. E. M. .465 * TOTAL 19.4410 9 1' _ 0.05 * * * MAXIMUM 6.100 * M * * * * * * * * * * * * * * A * * * * * * * * * * * k * * * * * * * * * * k # A A * * R * A * A * * * # * * * # # k * * h * * h h A R * * * # * * * * R * * * * * * * * * * * MINIMUM 1.600 * LEVENE'S TEST FOR EQUAI. VARIANCES 3, 6 1.21 .3664 * * * SAMPLE SIZE 10 * * * * * * * * * * * * A * * * * * * * * * A * A * k * * * * * * * A * * * * * * * * * * * R A * * * A * * h ir * A A A A * * A A h * * * A * * * * * F A * * * * * * h * A * * * * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITIIIN-GROUP * VARIANCES NOT ASSUMEI) TO BE EQLIAL * WELC11 3, 3 8.43 .0567 * BROl9N-FORSYTIIE 3, 3 5.49 .0978 a ti1GN11•'7C:A21'I'LY DIPI'I':RI:NT l•'ROM :iIIAM CON'PI2OL, :iTUUI:N'I`I'/,f;l) RAN('.I: 0.0') YcO9l.,IGBC/

Final Report 1-1729.005 TABLE CCH21 ANALYSIS OF RED BLOOD CELL COUNTS AFTER 3 WEEKS EXPOSURE HISTOGRAM OF RBC REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 11.100) 10.800) * 10.500) 10.200) * 9 .900) N * * 9.600)* N M 9.300) * N 9.000)* * * 8.700) 8.400) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 9.817 9.717 STD.DEV. .876 .544 R.E.S.D. .996 .631 S. E. M. .506 .314 MAXIMUM 10.790 10.130 MINIMUM 9.090 9.100 9.465 .276 .346 .195 9.660 9.270 9.315 .629 .789 .445 9.760 8.870 SAMPLE SIZE 3 3 2 2 ******************************* ********************k********** ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 9.616 * BETWEEN GROUPS .3780 3 .1260 .29 .8310 STD.DEV. .575 * WITHIN GROUPS 2.6008 6 .4335 p> 0.05; R.E.S.D. .589 S. E. M. .182 * TOTAL 2.9788 9 NOT SIGNIFICANT ************R**********k************iF******************************************************* MAXIMUM 10.790 MINIMUM 8.870 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.41 .3282 *******k************************************************************************************ SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 .23 .8681 * BROWN-FORSYTHE 3, 5 .34 .8004 sEOyz,sea

Final Report 1-1729.005 TABLE CCH22 ANALYSIS OF HEMOGLOBIN AFTER 3 WEEKS EXPOSURE HISTOGRAM OF HGB REF D6TEST D10TEST SHAM .................... ......+.......................+.......................+ MIDPOINTS 21.000) 20.000) 19.000) 18.000)* 17.000) * 16.000) M M M ].5.000) * * * M* 14.000) 13.000) 12.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WI TH *'S, N'S OTHERWISE MEAN 16.233 15.767 STD.DEV. 1.401 .971 R.E.S.D. 1.603 1.092 S. E. M. .809 .561. MAXIMUM 17.800 16.600 MINIMUM 15.100 14.700 SAMPLE SIZE 3 3 15.550 1.344 1.684 .950 16.500 14.600 2 15.100 .566 .709 .400 15.500 14.700 2 ALI. GROUPS COMBINED •************RR#*##RR#********* ANALYSIS OF VARIANCE TABLE *****R*k**R#k*#kk###**RR#**k*** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 15.730 * BETWEEN GROUPS 1.6227 3 .5409 .41 .7526 STD.DEV. 1.031 * WITHIN GROUPS 7.9383 6 1.3231 R.E.S.D. 1.052 * p > 0.05; S. E. M. .326 * TOTAL 9.5610 9 NOT SIGNIFICANT *********************************************************************************k********** MAXIMUM 17.800 MINIMUM 14.600 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 1.11 .4159 ************#***********R***********#***#**#**********************#******#******#*********** SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 .47 .7271 * BROWN-FORSYTHE 3, 4 .43 .7403 74OxGEiC1

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Final Report I-1729.005 TABLE CCH24 ANALYSIS OF NEUTROPHIL COUNTS AFTER 3 WEEKS EXPOSURE HISTOGRAM OF NEUTS REF D6TEST DIOTEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 45.000) 42.000)* 39.000) M* 36.000)N 33.000) * * 30.000) * 27.000) M* N 24.000) * 21.000) * 18.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 34.667 25.667 27.000 38.000 STD.DEV. 5.508 2.517 8.485 .000 R.E.S.D. 6.481 2.729 10.635 .000 S. E. M. 3.180 1.453 6.000 .000 MAXIMUM 41.000 28.000 33.000 38.000 MINIMUM 31.000 23.000 21.000 38.000 SAMPLE SIZE 3 3 2 2 ALL GROUPS COMBINED ***A**************#*******h**** * * * A * k * * * * * * * * * * * * * * * * * * * * * * * * * * ANALYSIS OF VARIANCE TABLE MEAN 31.100 * SOURCE BETWEEN GROUPS SUM OF SQUARES 255.5667 DF 3 MEAN SQUARE 85.1889 F VALUE 3.52 TAIL PROBABILITY .0888 STD.DEV. 6.674 * WITHIN GROUPS 145.3333 6 24.2222 R.E.S.D. 3.963 * p > 0.05; S. E. M. 2.111 * TOTAL 400.9000 9 NOT SIGNIFICANT **********************k*************A*****k**********.F***1.**#*******k*********1***R****k**** MAXIMUM 41.000 MINIMUM 21.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 8.67 .0133 ***********R*****f***********************A***************#*********R**#*****#********#A***** SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 1.65 .3135 * BROWN-FORSYTHE 3, 2 3.07 .2553 NZ.0.2r.fOa Cf

Final Report I-1729.005 TABLE CCH25 ANALYSIS OF LYMPHOCYTE COUNTS AFTER 3 WEEKS EXPOSURE HISTOGRAM OF LYMPHS REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 81.000) 78.000) 75.000) * 72.000) M 69.000) * * N 66.000) * 63.000)N * M* 60.000) 57.000) * 54.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 64.000 72.000 STD.DEV. 6.083 2.000 R.E.S.D. 7.163 2.047 S. E. M. 3.512 1.155 MAXIMUM 68.000 74.000 MINIMUM 57.000 70.000 SAMPLE SIZE 3 3 70.000 8.485 10.635 6.000 76.000 64.000 2 62.000 .000 .000 .000 62.On0 62.000 2 ALL GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE * * * * * * * * * * * * * * * * * * * * * * * * * * * k * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 67.200 * BETWEEN GROUPS 169.6000 3 56.5333 2.20 .1886 STD.DEV. 5.996 * WITHIN GROUPS 154.0000 6 25.6667 R.E.S.D. 3.963 * p > 0.05; S. E. M. 1.896 * TOTAL 323.6000 9 NuT SIGNII•'ICAN'P #************1#****************A***A*k**R*il**************#********A***it****#***#************* MAXIMUM 76.000 MINIMUM 57.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 9.29 .0113 *R*R****************k****k**************k***********************************k*************** SAMPLE SIZE 10 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 1.16 .4291 * BROWN-FORSYTHE 3, 2 1.97 .3548 E;EO94sed

Final Report I-1729.005 TABLE CCH26 ANALYSIS OF MONOCYTE COUNTS AFTER 3 WEEKS EXPOSURE HISTOGRAM OF MONO REF D6TEST U10TEST SHAM .......................+.......................+.......................+.......................+ VAR 9 EXCLUDED VALUES NOT REPORTED VALUES ** * ** TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE MIDPOINTS 2.100) 1.950) M 1.800) 1.650) 1.500) 1.350) 1.200) 1.050)M** M .900) .750) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S N'S OTHERWISE MEAN 1.000 2.000 1.000 .000 STD.DEV. .000 .000 .000 .000 R.E.S.D. .000 .000 .000 .000 S. E. M. .000 .000 .000 .000 MAXIMUM 1.000 2.000 1.000 .000 MINIMUM 1.000 2.000 1.000 .000 SAMPLE SIZE 3 1 1 0 ALL GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE ******#********k#***********R** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 1.200 * BETWEEN GROUPS .8000 2 .4000 .00 1.0000 STD.DEV. .447 * WITHIN GROUPS .0000 2 .0000 R.E.S.D. .000 * p > 0.05; S. E. M. .200 * TOTAL .8000 4 NOT SIGNIFICANT * 1 * * * MAXIMUM 2.000 * * # * * * * * * * R * * * * k * * # * * * * A * * * * * * k * * * * * * * * * * * * f * * * * * * * * * * rt * * f M * * * * * * * * * * * * * * * * i. * k * t * * * * * # * MINIMUM 1.000 * LEVENE'S TEST FOR EQUAL VARIANCES 2, 2 .00 1.0000 #*A*#************************************************************k************************** SAMPLE SIZE 5 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 2, 0 .00 .0000 * BROWN-FORSYTHE 2, 0 .00 .0000 OV0946eFi

Final Report 1-1729.005 TABLE CCH27 ANALYSIS OF RETICULOCYTE COUNTS AFTER 3 WEEKS EXPOSURE HISTOGRAM OF RETCTN REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 1.800) 1.650) * 1.500) ** M 1.350) * 1.200) N 1.050) M .900)N * * .750) .600) * .450) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN .933 1.267 STD.DEV. .416 .404 R.E.S.D. .478 .478 S. E. M. .240 .233 MAXIMUM 1.400 1.500 MINIMUM .600 .800 1.000 1.550 .141 .071 .177 .089 .100 .050 1.100 1.600 .900 1.500 SAMPLE SIZE 3 3 2 2 ALL GROUPS COMBINED ***k*************R************* ANALYSIS OF VARIANCE TABLE ****R************************** * * SOURCE SUM OF SQUARES DF MEAN SQUA RE F VALUE TAIL PROBABILITY * MEAN 1.170 * BETWEEN GROUPS .5427 3 .1809 1.55 .2951 STD.DEV. .371 * WITHIN GROUPS .6983 6 .1164 R.E.S.D. .286 * p > 0.05; S. E. M. .117 * TOTAL 1.2410 9 NOT SIGNIFICANT ******************************************************************************************** MAXIMUM 1.600 MINIMUM .600 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 6 2.94 .1207 SAMPLE SIZE 10 ***********1********************************************************************************* * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 6.68 .0765 * BROWN-FORSYTHE 3, 5 2.12 .2160 tVaXsea

Final Report I-172.9.005 TABLE CC1128 CLINICAL CHEMISTRY FOR 6 WEEK SACRIFICE OF MICE NO. LABEL CIG TYPE GLUCOSE mg/dl CHOL mg/dl TGA mg/dl CREAT mg/dl BUN mg/d1 URIC ACID mg/dl TOT PROT g/dl ALBUMIN g/dl A/G RAT10 ---- -------- ---------- ---------- ---------- ---------- ---------- ------- ---------- ---------- ---------- 1 1151 REF 210.700 86.700 112.800 .460 27.900 3.100 4.800 2.500 1.087 2 1152 REF 168.700 76.900 118.800 .320 27.400 2.700 4.400 2.400 1.200 3 1153 REF 260.300 82.300 114.800 .470 34.300 2.800 4.500 2.400 1.143 4 1163 D6 173.700 88.600 119.500 .340 26.200 2.800 4.700 2.300 .958 5 1164 D6 121.500 87.900 133.800 .380 24.600 2.100 4.600 2.300 1 6 1165 D6 191.200 89.600 124.200 .350 28.300 1.800 4.700 2.400 1.043 7 1175 D10 156.500 75.400 208.900 .710 22.500 2 4.700 2.400 1.043 8 1176 D10 195.800 75.700 132.500 .500 29.500 4 4.500 2.300 1.045 9 1177 D10 180.800 90 88.200 .350 24.700 .300 5.300 2.700 1.038 10 1186 SHAM 187.900 76.800 49.200 .400 19.700 1.200 4.900 2.500 1.042 11 1188 SHAM 215.500 82.600 42.900 .320 18.100 .500 4.600 2.500 1.190 12 1189 SHAM 208.900 87.600 43.200 .340 15.200 .400 4.500 2.400 1.143 C.VV 74Ci00

Final Report 1-1729.005 TABLE CCH29 CLINICAL CHEMISTRY FOR 6 WEEK SACRIFICE OF MICE NO. LABEL LDH U/1 ALKPHOS U/1 SGPT U/1 SGOT U/1 CALCIUM mg/dl PHOS mg/dl --------- ---------- ---------- ---------- ---------- 1 1151 152 70 51 128 9.600 7.600 2 1152 272 70 NRXa~ NRX 8.100 7.900 3 1153 117 65 40 112 10.300 8.300 4 1163 361 64 NRX NRX 8.500 7.200 5 1164 157 59 51 143 9 7.500 6 1165 215 59 55 195 8.600 8.600 7 1175 229 75 60 179 9.200 7.500 8 1176 401 52 NRX NRX 9.100 7.700 9 1177 103 68 NRX NRX 8.600 8.600 10 1186 195 63 NRX 187 8.800 7.400 11 1188 105 61 NRX 108 7.600 8.200 12 1189 138 56 NRX 68 8.700 8.900 aNRX =NO REACTION ABOVE BACKGROUND Etro~wsea

Final Report 1-1729.005 TARLE CCH30 HEMATOLAGY NOR 6 WEEK SACRINICE OF MICE NO. LABEL CIG TYPE WBC thousand RF3C million HG13 yl FICT $ NEUTS 2 LYMPIIS % MONOCYTE % RETCTN $ PLATELET ---- 1 -------- 1151 ---------- REE ---------- 6.200 ---------- 9.960 ---------- 16.900 ---------- 49.400 ---------- 26 ---------- 74 ----------- NIt' ---------- 4.100 ---------- 1 2 1152 REF 7.100 9.110 16.100 46.700 25 74 MP 3.100 1.600 3 1153 REF 7.700 8.270 15.600 41.600 17 82 1 6.800 1.700 4 1163 D6 8.300 9.460 16.700 47.200 21 76 rn< 4.700 1.400 5 1164 D6 6.100 9.470 16.800 46.900 29 68 Nk 5.400 1.500 6 1165 D6 24 9.680 16.300 49 28 71 NR 6.900 Nlt 7 1175 D10 7.500 9.690 17.200 48.100 52 38 N1< 1.100 1.500 8 1176 D10 5.700 9.830 16.800 48.300 24 74 N1< 3.400 1.100 9 1177 D10 14.600 8.160 17.200 41.300 31 68 NR 3.900 1.500 10 1186 SHAM 8.900 9.410 15 47.100 30 69 1 5.100 1.100 11 1188 SIIAM 7.300 9.860 18.100 49.100 41 59 NIZ 3.200 1.200 12 1189 SHAM 21.300 9.200 16.400 46.800 43 55 2 7.600 1.300 aNR = NUNE RI:PORTED OR ODSL•'ItVED. ~t~oy~ssa

Final Report 1-1729.005 TABLE CCH31 HEMATOLOGY FOR 6 WEEK SACRIFICE OF MICE NO. LABEL PMNCTN EOSIN $ 1 1151 -- NRa ---- NR 2 1152 NR 1.0 3 1153 NR NR 4 1163 NR 2.0 5 1164 NR 2.0 6 1165 NR 1.0 7 1175 NR NR 8 1176 NR 1.0 9 1177 NR 1.0 10 1186 NR NR 11 1188 NP, NR 12 1189 NR NR aNR = NOT REPORTED OR OBSERVED stroy1,Gsa

Final Report 1-1729.005 TABLE CCH32 ANALYSIS OF GLUCOSE VALUES AFTER 6 WEEKS EXPOSURE IIISTOGRAM OF GLUCOSE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOIN'PS 280.000) 260.000)* 240.000) 220.000)M * 200.000) * * M 180.000) * M * 160.000)* N * 140.000) 120.000) * 100.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 213.233 162.133 STD.DEV. 45.853 36.261 R.E.S.D. 48.165 41.581 S. E. M. 26.473 20.935 MAXIMUM 260.300 191.200 MINIMUM 168.700 121.500 SAMPLE SIZE 3 3 177.700 19.833 21.695 11.450 195.800 156.500 3 204.100 14.412 16.578 8.321 215.500 187.900 3 ALL GROUPS COMBINED ***************************x*** ANALYSIS OF VARIANCE TABLE ***********x******************* * * SOURCE SUM OF SQUARES OF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 189.292 * BETWEEN GROUPS 4993.2949 3 1664.4316 1.66 .2523 STD.DEV. 34.417 * WITHIN GROUPS 8036.7316 8 1.004.5915 R.E.S.D. 27.294 * p > 0.05; S. E. M. 9.935 * TOTAL 13030.0254 11 NOT SIGNIFICANT MAXIMUM 260.300 *k*t#************k******************k***********************k**********************k********* MINIMUM 121.500 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 1.22 .3621 SAMPLE SIZE 12 **************x*************************************k**********k**k******k****k************k * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 1.59 .3235 * BROWN-FORSYTHE 3, 5 1.66 .2897 9ttQ946Bc4

Final Report 1-1729.005 TABLE CCH33 ANALYSIS OF CHOLESTEROL AFTER 6 WEEKS EXPOSURE HISTOGRAM OF CHOL REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 92.000) 90.000) 88.000) 86.000)* 84.000) 82.000) M 80.000) 78.000) 76.000) * 74.000) GROUP MEANS ARE DENOTED BY M'S F * * M* * M N ** * THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 81.967 88.700 80.367 82.333 STD.DEV. 4.908 .854 8.344 5.405 R.E.S.D. 5.185 .921 9.858 5.662 S. E. M. 2.834 .493 4.817 3.121 MAXIMUM 86.700 89.600 90.000 87.600 MINIMUM 76.900 87.900 75.400 76.800 SAMPLE SIZE 3 3 3 3 ALL GROUPS COMBINED **********R*#****R*##****#*#R** *************k*k*************** ANALYSIS OF VARIANCE TABLE R * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 83.342 * BETWEEN GROUPS 121.4093 3 40.4698 1.31 .3369 STD.DEV. 5.790 * WITHIN GROUPS 247.3202 8 30.9150 R.E.S.D. 4.611 * p > 0.05; S. E. M. 1.671 * TOTAL 368.7294 11 NOT SIGNIFICANT MAXIMUM 90.000 **R**#RR*#kR#R#**#*********R**##*#R*#***RR*R*****R****#******************R*#*******#******** MINIMUM 75.400 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 2.87 .1039 SAMPLE SIZE 12 ******#****RR*#*R********#R**#******R****#*****#****#**********#*****#***A**************#*** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 2.89 .2030 * BROWN-FORSYTHE 3, 5 1.31 .3687 Z.Vos11sea

Final Report I-1729.005 TABLE CCH34 HISTOGRAM OF TGA ANALYSIS OF TRIGLYCERIDES AFTER 6 WEEKS EXPOSURE REF U6TL•:ST D L OTE ST SIIAM ....................... r....................... i....................... +....................... + MIDPOINTS 270.000) 240.000) 210.000) 180.000) 150.000) 120.000)M** 90.000) 60.000) 30.000) .000) * M* * * M k * `~GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE W1TH *'S, N'S OTHERWISE MEAN 115.467 STD.DEV. 3.055 R.E.S.D. 3.411 S. E. M. 1.764 MAXIMUM 118.800 MINIMUM 112.800 SAMPLE SIZE 3 ALI. GROUPS COMBINED MEAN 107.400 STD.DEV. 47.010 R.E.S.D. 17.694 S. E. M. 13.571 MAXIMUM 208.900 MINIMUM 42.900 SAMPLE SIZE 12 125.833a 143.200a 7.289 61.057 8.152 67.233 4.208 35.251 133.800 208.900 119.500 88.200 3 3 *********k*******#*k*******AA** ANALYSIS * * SOURCE SUM OF SQUARES * BETWEEN GROUPS * WITNIN GROUPS * * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS L'OR WITHIN-GROUP * VARIANCES NOT ASSUMEI) TO BE EQUAI. * WELCII * BROWN-FORSYTHE 45.100 3.554 4.196 2.052 49.200 42.900 3 OF VARIANCE TABLE R * * !F * * * * * * * * * * * * * * k * * * * * * * * * * * k UE MEAN SQUARE L' VALUE TAIL PROI3ABILI'1'Y 16703.3672 3 5567.7891 5.86 .0204 7606.1527 8 950.7691 * TOTAI. 24309.5195 ll N<_ 0.05 * * * * * * * * * * * * * * * * # k * * A * A * * * * * * A * * * * * * * * * k * * * * * * * * * * * * * * k * * * * * k * * A * * k * * * * * * * A * * * * * * * * * * * * * * * * * * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 5.68 .0221 * * * * * * * * * * * * * * * * * * * * * * * # * * * * * * * * * * * * * * * * * * * * * * * * A * * * # * * * * l. * * A A A * * * 4 * * * * * A A * A * * * * * * * * A * * * * * * !. 3, 4 191.82 .0001 3, 2 5.86 .1493 aS1GNl1•'ICANTI.Y DIFFI~;ItI'N'P FROM SIIAM CON'PROL, 5'I'IIDI:N'I'IGI?Il P F ` 0.05. 8W94,GeR

Final Report 1-1729.005 TABLE CCH35 ANALYSIS OF CREATININE AFTER 6 WEEKS EXPOSURE IIISTOGRAM OF CREAT MIDPOINTS REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ .750) .700) .650) .600) * .550) .500) .450) ** M .400) N R .350) M* .300) * * M * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S , N'S OTHERWISE MEAN .417 .357 STD.DEV. .084 .021 R.E.S.D. .099 .024 S. E. M. .048 .012 MAXIMUM .470 .380 MINIMUM .320 .340 .520 .353 .181 .042 .194 .048 .104 .024 .710 .400 .350 .320 SAMPLE SIZE 3 3 3 3 ALL GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE *********************R********* * * SOURCE SUM OF SQUARES DF MEAN SQUA RE F VALUE TAIL PROBABILITY * MEAN .412 * BETWEEN GROUPS .0546 3 .0182 1.74 .2367 STD.DEV. .112 * WITHIN GROUPS .0838 8 .0105 H.E.S.D. .078 * p > 0. 05; S. E. M. .032 * TOTAL .1384 11 NOT SIGNIFICANT MAXIMUM .710 ****R*****R****#*********************************************************k**k*************** MINIMUM .320 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 2.97 .0969 SAMPLE SIZE 12 #R*R**A*************####*k****R************R***R****#******************kk*t1*************A*** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .96 .4940 * BROWN-FORSYTHE 3, 3 1.74 .3308 E,V0946eg

TABLE CCH36 ANALYSIS OF BLOOD UREA NITROGEN AFTER 6 WEEKS EXPOSURE IIISTOGRAM OL•' BUN Final Report 1-1729.005 RF.L' D6TE:S'P 1)1UTEST SIIAM .......................+........................ 1....................... +........................ h1IDPOINTS 36.000) 34.000) * 32.000)N * M* 30.000)** M 28.000) * 26.000) 24.000) 22.000) M 20.000) * 1.8.000) `'CROUI' MEANS ARE DENO'l'EU BY M'S Il THEY COINCIDE WITH *'S, N'S OTHE.IfWI SE MEAN 31.750d 30.533 a 31.550' 21.350 S'l'f).DE:V. 1.953 2.238 .283 1.202 R.E.S.I). 2.302 2.422 .354 1.507 S. L'. M. 1.127 1.292 .200 .850 MAXIMUM 34.000 32.900 31.750 22.200 MINIMUM 30.500 28.450 31.350 20.500 SAMPLE S1lE: 3 3 2 2 ALI. GROUPS COM6INEl) i. * * * * * * * * * A k * * * * * * * * * A * k * k * * * * * ANALYSIS A * * * * * OF VARIANCI: 'I'AIiL,E * * A A * * * A * * A A k * * * * A * * * * * k * * * SOURCE SUM OF SQUARES DE' MEAN Ei(_)L1ARE E' VALUE '1'AIL. PROBAI3II,ITY * MEAN 29.265 * BETWEEN CROIIPS 159.0886 3 53.0295 16.60 .0026 S'1'D.DEV. 4.450 * WITHIN GROUPS 19.1717 6 3.1953 R.E.S.D. 1.497 * S. E. M. 1.407 * TO'l'AL 178.2602 9 1) 0.05 MAXIMUM 34.000 * * * k * * * * k * A * * * A * A * A * * A k A * * A A * * * A A * k * A * * A A * * * * A A * * * R A * A A * k A A * A k * A * * A * * * k k * A k * A * A A A * * k A * A k A A * k MINIMUM 20.500 * LEVhiNE'S TES'1' FOR EQUAL VARIANCES 3, 6 1.69 .2672 SAMPI.E; S f'LL; 10 * * k * * A * * * * A * A * A A * * A k A A * k k * * * k * * * A * * * * k * * * * k A A A * A * * * * * * A k * A R A A A A * * A A * * * * * * * A * * * A A A * k A * * * * * * A * * ONE-WAY ANALYSIS UL' VARIANCE * TEST STATISTICS FOR WITHIN-GROl1P * VARIANCES NO'l' ASSLIMEU TO 13E EQUAL * WELCH 3, 3 30.51 .0095 * IIROWN-FORSYTHE 3, 5 21.51 .0028 `)5IGN11•'LCANTLY DIl•'l'ERICNT FROM :.i11AM CON'1'ROL, S'l'lJI)KN'I'1'I.I;D RAN(;I: '1'I;ST, I' < 0.05. OS09G66R

Final Report 1-1729.005 TABLE CCH37 HISTOGRAM OF ORICACID ANALYSIS OF URIC-ACID AFTER 6 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 7.000) 6.000) 5.000) 4.000) 3.000)M** * * 2.000) M* M 1.000) .000) -1.000) -2.000) * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 2.867 STD.DEV. .208 R.E.S.D. .239 S. E. M. .120 MAXIMUM 3.100 MINIMUM 2.700 SAMPLE SIZE 3 ALL GROUPS COMBINED MEAN 1.975 STD.DEV. 1.182 R.E.S.D. .698 S. E. M. .341 MAXIMUM 4.000 MINIMUM .300 SAMPLE SIZE 12 2.233 2.100 .513 1.852 .580 1.944 .296 1.069 2.800 4.000 1.800 .300 3 3 *****************#************* * M ** .700 .436 .512 .252 1.200 .400 3 ANALYSIS OF VARIANCE TABLE * * * * * * * * * * * * * * * * * * * k * * * * * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS 7.5092 3 2.5031 2.55 .1289 * WITHIN GROUPS 7.8533 8 .9817 * p > 0. 05; * TOTAL 15.3625 11 NOT SIGNIFICANT f*************************************#****R*********A********************************k***** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 2.70 .1161 *A*A****************************************************************************kk********** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 15.18 .0119 * BROWN-FORSYTHE 3, 3 2.55 .2312 IS0946et?

Final Report 1-1729.005 TABLE CCH38 ANALYSIS OF TOTAL PROTEIN AFTER 6 WEEKS EXPOSURE HISTOGRAM OF TOTPROT REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 5.550) 5.400) 5.250) * 5.100) 4.950) 4.800)* N 4.650) M** * M 4.500)M * * 4.350)* 4.200) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 4.567 4.667 STD.DEV. .208 .058 R.E.S.D. .239 .068 S. E. M. .120 .033 MAXIMUM 4.800 4.700 MINIMUM 4.400 4.600 SAMPLE SIZE 3 3 4.833 .416 .478 .240 5.300 4.500 3 4.667 .208 .239 .120 4.900 4.500 3 ALL GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL. PROBABILITY * MEAN 4.683 * BETWEEN GROUPS .1100 3 .0367 .56 .6580 STD.DEV. .241 * WITHIN GROUPS .5267 8 .0658 P > 0 05; R.E.S.D. .218 S. E. M. .069 * TOTAL .6367 11 . NOT SIGNII•'ICANT MAXIMUM 5.300 ******************************************************************************************** MINIMUM 4.400 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 3.39 .0745 SAMPLE SIZE 12 ***************************i#********************************************************k**k**** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .28 .8363 * BROWN-FORSYTHE 3, 4 .56 .6708 9V,71.160IR

Final Report 1-1729.005 TABLE CCH39 ANALYSIS OF ALBUMIN AFTER 6 WEEKS EXPOSURE HISTOGRAM OF ALBUMIN REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 2.750) 2.700) * 2.650) 2.600) 2.550) 2.500) * ** 2.450)N N N 2.400)** * * * 2.350) N 2.300) ** * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 2.433 2.333 STD.DEV. .058 .058 R.E.S.D. .068 .068 S. E. M. .033 .033 MAXIMUM 2.500 2.400 MINIMUM 2.400 2.300 2.467 .208 .239 .120 2.700 2.300 2.467 .058 .068 .033 2.500 2.400 SAMPLE SIZE 3 3 3 3 ***********************t***k*** ALL GROUPS COMBINED ******************************* ANALVSIS OF VARIANCE TABLE * * SOURCE SUM OF S QUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 2.425 * BETWEEN GROUPS .0 358 3 .0119 .90 .4842 STD.DEV. .114 * WITHIN GROUPS .1 067 8 .0133 R.E.S.D. .095 . P > 0.05; S. E. M. .033 * TOTAL .1 425 11 NOT SIGNIFICANT ****************************************************************************************1*** MAXIMUM 2.700 MINIMUM 2.300 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 4.55 .0386 ***************************************************M***************k************************ SAMPLE SIZE 12 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 2.26 .2236 * BROWN-FORSYTHE 3, 3 .90 .5350 Esoy4sea

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TABLE CCH41 HISTOGRAM OF LDH ANALYSIS OF LACTIC DEHYDROGENASE AFTER 6 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 440.000) 400.000) 360.000) 320.000) 280.000)* * * 240.000) N M 200.000)N * * 160.000)* * N 120.000)* * ** 80.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 180.333 STD.DEV. 81.292 R.E.S.D. 93.805 S. E. M. 46.934 MAXIMUM 272.000 MINIMUM 117.000 SAMPLE SIZE 3 ALL GROUPS COMBINED MEAN 203.750 STD.DEV. 98.048 R.E.S.D. 90.324 S. E. M. 28.304 MAXIMUM 401.000 MINIMUM 103.000 SAMPLE SIZE 12 244.333 244.333 105.116 149.591 119.388 160.321 60.689 86.366 361.000 401.000 157.000 103.000 3 3 *********A********1fi*R********I** * 146.000 45.530 50.143 26.287 195.000 105.000 3 ANALYSIS OF VARIANCE TABLE Final Report I-1729.005 * * * * * * * * k * k * * * * * * * * * k * * * * * * * k * * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS 21532.2500 3 7177.4170 .68 .5876 * WITHIN GROUPS 84216.0000 8 10527.0000 * p > 0.05; * TOTAL 105748.2500 11 NOT SIGNIFICANT * * * * * * * * * * * * * * * * * * * * * * * * * * * * k * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * k * * ~r * * * * * * * * * * * * * * * * * * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 1.15 .3874 *L************************************k***k****k******************k*****************k******* * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 * BROWN-FORSYTHE 3, 5 .78 .5616 .68 .6002 SS09L68d

Final Report 1-1729.005 TABLE CCH42 HISTOGRAM OF ALKPHOS ANALYSIS OF ALKALINE PHOSPHATASE AFTER 6 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 78.000) 75.000) 72.000) 69.000)M* * * 66.000)* N 63.000) 60.000) 57.000) 54.000) 51.000) * M* * M * * GROUP MEANS ARE DENOTED ElY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 68.333 STD.DEV. 2.887 R.E.S.D. 3.411 S. E. M. 1.667 MAXIMUM 70.000 MINIMUM 65.000 SAMPLE SIZE 3 ALL GROUPS COMBINED MEAN 63.500 STD.DEV. 6.571 R.E.S.D. 5.163 S. E. M. 1.897 MAXIMUM 75.000 MINIMUM 52.000 SAMPLE SIZE 12 60.667 65.000 60.000 2.887 11.790 3.606 3.411 13.303 4.093 1.667 6.807 2.082 64.000 75.000 63.000 59.000 52.000 56.000 3 3 3 ***********R*R*******RR*****R** ANALYSIS OF VARIANCE TABLE ******************************* R * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS 137.6667 3 45.8889 1.09 .4080 * WITHIN GROUPS 337.3333 8 42.1667 * p > 0.05; * TOTAL 474.9999 11 NOT SIGNIFICANT ***********R*****R************************************************************************** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 3.91 .0545 ************************************************x******************************************* * ONE-WAY ANALYSIS OF' VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 * BROWN-FORSYTHE 3, 3 3.60 .1237 1.09 .4731 9SQ!jt,68tj

Final Report I-172A.005 TABLE CCH43 ANALYSIS OF SGPT AFTER 6 WEEKS EXPOSURE HISTOGRAM OF SGPT REF D6TEST D10TEST SIIAM ....................... 1........................................................................ .......+ VAR 14 EXCLUDED VALUES QUANTITY NOT SUFFICIENT FOR ANALYSIS * * * * * * * TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE MIDPOINTS 63.000) 60.000) 57.000) 54.000) M 51.000)* 48.000) 45.000)N 42.000) 39.000)* 36.000) M * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 45.500 53.000 STD.DEV. 7.778 2.828 R.E.S.D. 9.748 3.545 S. E. M. 5.500 2.000 MAXIMUM 51.000 55.000 MINIMUM 40.000 51.000 SAMPLE SIZE 2 2 60.000 .000 .000 .000 60.000 60.000 1 .000 .000 .000 .000 .000 .000 0 ALL GROUPS COMBINED **#A***A***AAA**A#*******A***** ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 51.400 * BETWEEN GROUPS 148.7000 2 74.3500 2.17 .3154 STD.DEV. 7.369 * WITHIN GROUPS 68.5000 2 34.2500 R.E.S.D. 4.204 * p > 0.05; S. E. M. 3.295 * TOTAL 217.2000 4 NOT SIGNIFICANT MAXIMUM 60.000 AkA*#**AA******#A#AAA**AA#iA#AA##k**#*****kAA**A*#A************AA**#A******************#**** MINIMUM 40.000 * LEVENE'S TEST FOR EQUAL VARIANCES 2, 2 .00 1.0000 SAMPLE SIZE 5 **A**#**A**********#**#A***AA**#*#*******iF***#***k**********#****#***********#****A****ik**** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 2, 3 .39 .7060 * BROWN-FORSYTHE 2, 1 3.62 .3485 4SQ946BR

Final Report 1-1729.005 TABLE CCH44 ANALYSIS OF SGOT AFTER 6 WEEKS EXPOSURE HISTOGRAM OF SGOT REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ VAR 15 EXCLUDED VALUES MIDPOINTS QUANTITY NOT SUFFICIENT FOR ANALYSIS * * ** TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE 220.000) 200.000) * 180.000) M * 160.000) N 140.000) * 120.000)M* N 100.000) * 80.000) 60.000) * 40.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 120.000 169.000 179.000 121.000 STD.DEV. 11.314 36.770 .000 60.556 R.E.S.D. 14.180 46.084 .000 67.540 S. E. M. 8.000 26.000 .000 34.962 MAXIMUM 128.000 195.000 179.000 187.000 MINIMUM 112.000 143.000 179.000 68.000 SAMPLE SIZE 2 2 1 3 ALL GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE ******************************* R * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 140.000 * BETWEEN GROUPS 5086.0000 3 1695.3333 .77 .5681 STD.DEV. 44.561 * WITHIN GROUPS 8814.0000 4 2203.5000 R.E.S.D. 33.496 * p > 0.05; S. E. M. 15.755 * TOTAL 13900.0000 7 NOT SIGNIFICANT MAXIMUM 195.000 ****************R***************R**k****************it*************************************** MINIMUM 68.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 4 2.00 .2565 SAMPLE SIZE 8 **********************************************************~k#********************k**#*#****** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .80 .5538 * BROWN-FORSYTHE 3, 3 1.50 .3745 asoyW;et4

Final Report 1-1729.005 TABLE CCH4S ANALYSIS OF CALCIUM AFTER 6 WEEKS EXPOSURE HISTOGRAM OF CALCIUM REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 10.800) 10.400)* 10.000) 9.600) * 9.200) N ** 8.800) M N ** 8.400) ** * N 8.000) * 7.600) * 7.200) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 9.333 8.700 STD.DEV. 1.124 .265 R.E.S.D. 1.262 .307 S. E. M. .649 .153 MAXIMUM 10.300 9.000 MINIMUM 8.100 8.500 SAMPLE SIZE 3 3 8.967 .321 .375 .186 9.200 8.600 3 8.367 .666 .785 .384 8.800 7.600 3 ALL GROUPS COMBINED ******************************* ***************/.*************** ANALYSIS OF VARIANCE TABLE * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 8.842 * BETWEEN GROUPS 1.5092 3 .5031 1.07 .4145 STD.DEV. .692 * WITHIN GROUPS 3.7600 8 .4700 R. E:.S.D. .582 * p > 0.05; S. E. M. .200 * TOTAL 5.2692 11 NOT SIGNIFICANT MAXIMUM 10.300 #***k**************************************************************************k************ MINIMUM 7.600 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 3.04 .0924 SAMPLE SIZE 12 ******************#************************************************************************* * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .80 .5558 * BROWN-FORSYTHE 3, 4 1.07 .4556 GS094668

Final Report 1-1729.005 TABLE CCI146 ANALYSIS OF PHOSPHORUS AFTER 6 WEEKS EXPOSURE fIISTOGRAM OF PHOS REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 9.300) 9.000) 8.700) * 8.400) * 8.100) M 7.800)M N M 7.500) * * * * 7.200) * 6.900) 6.600) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 7.933 7.767 STD.DEV. .351 .737 R.E.S.D. .375 .853 S. E. M. .203 .426 MAXIMUM 8.300 8.600 MINIMUM 7.600 7.200 SAMPLE SIZE 3 3 7.933 .586 .682 .338 8.600 7.500 3 8.167 .751 .785 .433 8.900 7.400 3 ALL GROUPS COMBINED ******************************* *************#****k************ ANALYSIS OF VARIANCE 'fABLE * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE 'PAIL PROBABILITY * MEAN 7.950 * BETWEEN GROUPS .2433 3 .0811 .21 .8893 STD.DEV. .555 * WITHIN GROUPS 3.1467 8 .3933 R.E.S.D. .575 * p > 0.05; S. E. M. .160 * TOTAL 3.3900 11 NOT SIGNIFICANT MAXIMUM 8.900 #********#*R**************##*##********k#****#****#*#*##*#*****###********#**********k****** MINIMUM 7.200 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 .68 .5877 SAMPLE SIZE 12 *******#*#*****#******##**#***#*****#**#***#*****#***#*****#*#**************#**k******k***** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .11 .9488 * BROWN-FORSYTHE 3, 7 .21 .8889 0Jl1 9f.4O H k3

Final Report 1-1729.005 TABLE CCH47 ANALYSIS OF WHITE BLOOD CELL COUNTS AFTER 6 WEEKS EXPOSURE HISTOGRAM OF WBC REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 27.000) 24.000) * 21.000) * 18.000) 15.000) * 12.000) N N 9.000)* * N * 6.000)M* * ** * 3.000) .000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 7.000 12.800 STD.DEV. .755 9.762 R.E.S.D. .819 11.461 S. E. M. .436 5.636 MAXIMUM 7.700 24.000 MINIMUM 6.200 6.100 SAMPLE SIZE 3 3 9.267 4.706 5.458 2.717 14.600 5.700 3 12.500 7.663 9.005 4.424 21.300 7.300 3 **#*#***#*k#******#*******#**#* ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 10.392 * BETWEEN GROUPS 69.0425 3 23.0142 .52 .6797 STD.DEV. 6.197 * WITHIN GROUPS 353.4467 8 44.1808 R.E.S.D. 5.702 * p > 0.05; S. E. M. 1.789 * TOTAL 422.4891 11 NOT SIGNIFICANT ****#*********************'***********#****#*****#***#*************#*#***#******##********##* MAXIMUM 24.000 MINIMUM 5.700 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 4.93 .0317 ***********************##***#*********#*R#**#****i****************#**#***###*#*A#*##***#**** SAMPLE SIZE 12 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 3 .77 .5842 * BROWN-FORSYTHE 3, 5 .52 .6863 1JII.J(„tGp O

Final Report 1-1729.005 TABLE CCH48 ANALYSIS OF RED BLOOD CELL COUNTS AFTER 6 WEEKS EXPOSURE HISTOGRAM OF RBC REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 10.500) 10.200) k * 9.900) * 9.600) M** * N 9.300) N ** 9.000)M 8.700) 8.400)* 8.100) 7.800) * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N' S OTHERWISE MEAN 9.113 9.537 STD.DEV. .845 .124 R.E.S.D. .866 .147 S. E. M. .488 .072 MAXIMUM 9.960 9.680 MINIMUM 8.270 9.460 SAMPLE SIZE 3 3 9.227 .926 1.092 .535 9.830 8.160 3 9.490 .337 .379 .195 9.860 9.200 3 * * * * * * * * * * * * * ALL GROUPS COMBINED * * * * * * * * * * * * * * * * * * ANALYSIS OF VARIANCE TABLE ***k******************k******** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 9.342 * BETWEEN GROUPS .3762 3 .1254 .29 .8283 STD.DEV. .586 * WITHIN GROUPS 3.4028 8 .4253 R.E.S.D. .529 * p > 0.05; S. E. M. .169 * TOTAL 3.7790 11 NOT SIGNIFICANT ******************************************************************************************** MAXIMUM 9.960 MINIMUM 8.160 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 2.67 .1189 k******************************************************************************************* SAMPLE SIZE 12 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .26 .8500 * BROWN-FORSYTHE 3, 5 .29 .8282 (.7V 0QCt .7L4

Final Report 1-1729.005 TABLE CCH49 ANALYSIS OF HEMOGLOBIN AFTER 6 WEEKS EXPOSURE HISTOGRAM OF HGB REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 21.000) 20.000) 19.000) 18.000) 17.000)* M* M** 16.000)M* * M 15.000) * 14.000) 13.000) 12.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *' S, N'S OTHERWISE MEAN 16.200 16.600 STD.DEV. .656 .265 R.E.S.D. .716 .307 S. E. M. .379 .153 MAXIMUM 16.900 16.800 MINIMUM 15.600 16.300 SAMPLE SIZE 3 3 17.067 .231 .273 .133 17.200 16.800 3 16.500 1.552 1.637 .896 18.100 15.000 3 ALE. GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE *******kk********************** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 16.592 * BETWEEN GROUPS 1.1625 3 .3875 .52 .6784 STD.DEV. .803 * WITHIN GROUPS 5.9267 8 .7408 R.E.S.D. .625 * p > 0.05; S. E. M. .232 * TOTAL 7.0892 11 NOT SIGNIFICANT ******************************************************************************************** MAXIMUM 18.100 MINIMUM 15.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 2.50 .1335 *************************************************************************i.****************** SAMPLE SIZE 12 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 2.09 .2444 * BROWN-FORSYTHE 3, 3 .52 .6960 E9U9&63R

Final Report I-1729.005 TABLE CCH50 ANALYSIS OF HEMATOCRIT VALUES AFTER 6 WEEKS EXPOSURE HISTOGRAM OF HCT REF ' D6TEST DIUTEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 50.000) 49.000) * * * 48.000) N ** N 47.000) * ** ** 46.000)N N 45.000) 44.000) 43.000) 42.000) * 41.000) R GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 45.900 47.700 45.900 47.667 STD.DEV. 3.961 1.136 3.985 1.250 R.E.S.D. 4.400 1.330 4.707 1.467 S. E. M. 2.287 .656 2.301 .722 MAXIMUM 49.400 49.000 48.300 49.100 MINIMUM 41.600 46.900 41.300 46.800 SAMPLE SIZE 3 3 3 3 ALL GROUPS COMBINED ****R*****R*#***###**#***#**R** ******************************* ANALYSIS OF VARIANCE TABLE M * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 46.792 * BETWEEN GROUPS 9.5425 3 3.1808 .37 .7772 STD.DEV. 2.670 * WITHIN GROUPS 68.8467 8 8.6058 R.E.S.D. 2.538 * P > 0.05; S. E. M. .771 * TOTAL 78.3891 11 NOT SIGNIFICANT MAXIMUM 49.400 *****************************************R*****k******k*****************k******************* MINIMUM 41.300 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 3.09 .0895 SAMPLE SIZE 12 **********RR**R#R**R#R*#**R****#*k************##*****#********k**#*k*k#*A*f*#kkk*##*k**k*k** * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 .28 .8380 * BROWN-FORSYTHE 3, 5 .37 .7789 V9Q946gR

Final Report 1-1729.005 TABLE CCHS1 ANALYSIS OF NEUTROPHIL COUNTS AFTER 6 WEEKS EXPOSURE HISTOGRAM OF NEUTS REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 55.000) 50.000) * 45.000) * 40.000) M 35.000) N 30.000) ** * * 25.000)M* N * 20.000) # 15.000) * 10.000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 22.667 26.000 35.667 38.000 STD.DEV. 4.933 4.359 14.572 7.000 R.E.S.D. 5.799 5.117 16.714 8.187 S. E. M. 2.848 2.517 8.413 4.041 MAXIMUM 26.000 29.000 52.000 43.000 MINIMUM 17.000 21.000 24.000 30.000 SAMPLE SIZE 3 3 3 3 ALL GROUPS COMBINED ******************************* ANALYSIS OF VARIANCE TABLE ******************************* # * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 30.583 * BETWEEN GROUPS 493.5834 3 164.5278 2.16 .1707 STD.DEV. 10.013 * WITHIN GROUPS 609 3333 8 76 1667 R.E.S.D. 7.636 * . . p > 0.05; S. E. M. 2.891 * TOTAL 1102.9165 11 NOT SIGNIFICANT MAXIMUM 52.000 ##**#****#*#*1t#******fl*******!e**i!***1.*******#******ir*#*********#*#*k#*A#*!.!***************** MINIMUM 17.000 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 3.15 .0864 SAMPLE SI # * ZE 12 * * t~ * fi # * # * # # # # * # * i1 * * * * fe # * # # * * * # * f' * # iF # h fr 1' * * .Y # * * * * # i. # /e * # * # # # 1r * * * * * * * * f. * * * tF * * * * # # k * * * * * * * * * * * * * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 2.77 .1750 * BROWN-FORSYTHE 3, 4 2.16 .2354 ssos4sea

Final Report 1-1729.005 TABLE CCH52 HISTOGRAM OF LYMPHS ANALYSIS OF LYMPHOCYTE COUNTS AFTER 6 WEEKS EXPOSURE REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ MIDPOINTS 91.000) 84.000) * 77.000)M* 70.000) 63.000) 56.000) 49.000) 42.000) 35.000) 28.000) * M* * * N * GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 76.667 STD.DEV. 4.619 R.E.S.D. 5.458 S. E. M. 2.667 MAXIMUM 82.000 MINIMUM 74.000 SAMPLE SIZE 3 ALL GROUPS COMBINED MEAN 67.333 STD.DEV. 11.765 R.E.S.D. 8.654 S. E. M. 3.396 MAXIMUM 82.000 MINIMUM 38.000 SAMPLE SIZE 12 71.667 60.000 4.041 19.287 4.434 22.513 2.333 11.136 76.000 74.000 68.000 38.000 3 3 ******************************* * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH * BROWN-FORSYTHE 61.000 7.211 8.187 4.163 69.000 55.000 3 ANALYSIS OF VARIANCE TABLE ******************************* * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * * BETWEEN GROUPS 599.3334 3 199.7778 1.73 .2377 * WITHIN GROUPS 923.3333 8 115.4167 * p > 0.05; * TOTAL 1522.6665 11 NOT SIGNIFICANT *****#**#******#**#*****##**##*****##*****##*****#**#**#******##*k#****k***#****A*****#*k*** * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 5.53 .0237 *#*****k*##******#******k#**k****#***#*****##***********************k**#******#************* 3, 4 2.83 .1704 3, 3 1.73 .3317 99Oy4fgd

Final. Report I-17?9.005 TABLE CCH53 ANALYSIS OF MONOCYTE COUNTS AFTER 6 WEEKS EXPOSURE HISTOGRAM OF MONO REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ VAR 9 EXCLUDED VALUES VALUES NOT REPORTED ** *** *** * MIDPOINTS 2.100) TABULATIONS AND COMPUTATIONS WHICH FOLLOW EXCLUDE VALUES LISTED ABOVE 1.950) * 1.800) 1.650) 1.500) N 1.350) 1.200) 1.050) M * .900) .750) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE MEAN 1.000 .000 STD.DEV. .000 .000 R.E.S.D. .000 .000 S. E. M. .000 .000 MAXIMUM 1.000 .000 MINIMUM 1.000 .000 SAMPLE SIZE 1 0 .000 .000 .000 .000 .000 .000 0 1.500 .707 .886 .500 2.000 1.000 2 *****k************************* ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE *******k*********************** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 1.333 * BETWEEN GROUPS .1667 1 .1667 .33 .6667 STD.DEV. .577 * WITHIN GROUPS .5000 1 .5000 R.E.S.D. .512 * p > 0.05; S. E. M. .333 * TOTAL .6667 2 NOT SIGNIFICANT ****!************************************i******k******************************************* MAXIMUM 2.000 MINIMUM 1.000 * LEVENE'S TEST FOR EQUAL VARIANCES 1, 1 .00 1.0000 *********************************************k************************k***********k********* SAMPLE SIZE 3 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 1, 0 .00 .0000 * BROWN-FORSYTIIE 1, 1 1.00 .5000 1_.9U !)4E',Bg

TABLE CCH54 ANALYSIS OF RETICULOCYTE COUNTS AFTER 6 WEEKS EXPOSURE HISTOGRAM OF RETCTN MIDPOINTS REF D6TEST D10TEST SHAM .......................+.......................+.......................+.......................+ 9.000) 8.000) * 7.000)* * 6.000) N 5 .000) N ** M 4.000)* * 3.000)* M * 2.000) 1.000) * .000) GROUP MEANS ARE DENOTED BY M'S IF THEY COINCIDE WITH *'S, N'S OTHERWISE Final Report 1-1729.005 MEAN 4.667 5.667 STD.DEV. 1.914 1.124 R.E.S.D. 2.183 1.262 S. E. M. 1.105 .649 MAXIMUM 6.800 6.900 MINIMUM 3.100 4.700 SAMPLE SIZE 3 3 2.800 1.493 1.740 .862 3.900 1.100 3 5.300 2.207 2.354 1.274 7.600 3.200 3 ****************************#** ALL GROUPS COMBINED ANALYSIS OF VARIANCE TABLE ********A********************** * * SOURCE SUM OF SQUARES DF MEAN SQUARE F VALUE TAIL PROBABILITY * MEAN 4.608 * BETWEEN GROUPS 14.6158 3 4.8719 1.62 .2599 STD.DEV. 1.875 * WITHIN GROUPS 24.0533 8 3.0067 R.E.S.D. 1.607 * p > 0.05; S. E. M. .541 * TOTAL 38.6692 11 NOT SIGNIFICANT *******************************************************************A****/.*****#************* MAXIMUM 7.600 MINIMUM 1.100 * LEVENE'S TEST FOR EQUAL VARIANCES 3, 8 .48 .7036 ************************************R*******************************1.****k****************** SAMPLE SIZE 12 * ONE-WAY ANALYSIS OF VARIANCE * TEST STATISTICS FOR WITHIN-GROUP * VARIANCES NOT ASSUMED TO BE EQUAL * WELCH 3, 4 1.86 .2774 * BROWN-FORSYTHE 3, 7 1.62 .2692 N90y4699

Final Report I-1729.005 X. REFERENCES Bertalanffy, F.D. Tritiated Thymidine Versus Colchicine Technique in the Study of Cell Population Cytodynamics. Lab. Investigation, 13:871-884, 1964. Burke, M.D., and Mayer, R.T. Ethoxyresorufin: Direct Fluorometric Assay of a Microsomal-O-Dealkylation Which Is Preferentially Inducible By 3-Methylcholanthrene. Drug Metab. Disp. 2:583, 1974. Coles, E.H. Veterinary Clinical Pathology. Third Edition. Philadelphia: W.B. Saunders Co., 1980. Dixon, W.J., and Brown, M.B. BMDP-79, Biomedical Computer Programs, P-Series. Berkeley: University of California Press, 1979. Dungworth, D.L., Schwartz, L.W., Tyler, W.S. and Phalen, R.F. Morphological Methods for Evaluation of Pulmonary Toxicity in Animals. Ann. Rev. Pharmacol. Toxicol. 16:381-399, 1976. Guerin, M.R., Stokely, J.R., Higgins, C.E., Moneyhun, J.H., and Holmberg, R.W. Inhalation Bioassay Chemistry - Walton Horizontal Smoking Machine for Inhalation Exposure of Rodents to Cigarette Smoke. J. Natl. Can. Inst., 63:441-448, 1979. Henry, C.J., Whitmire, C.E., Lopez, A., Dansie, D.R., Avery, M. D., Caton, J.E., Stokely, J.R., Holmberg, R.W., Guerin, M.R., and Kouri, R.E. The Dosimetry and Distribution of Whole Cigarette Smoke Particulates in Inbred Strains of Mice: Comparison of a Large Smoke- Exposure Machine (SEM) with a Small Capacity Smoke Exposure Machine (Walton). In: Pulmonary Toxicology of Respirable Particles, C.L. Sanders, F.T. Cross, G.E. Dagle, and J.A. Mahaffey (Eds.), Technical Information Center, U.S. Department of Energy, NTIS, pp. 177-192, 1980. Kanagalingam, K.K., Banerjee, A., Reed, S.M., Dansie, D.R., Benedict, W.F., Kouri, R.E., and Henry, C.J. Cigarette Smoke Increases DNA Replication in Lung and Sister Chromatid Exchange (SCE) in Bone Marrow of Mice. Proc. of Am. Assoc. Can. Res., 23:56, 1982. 23

Final Report I-1729.005 Kouri, R.E., and Nebert, D.W. Genetic Regulation of Susceptibility to Polycyclic Hydrocarbon-Induced Tumors in the Mouse. In: Origins of Human Cancer, H.H. Hiatt, J.D. Watson, and J.A. Winsten, eds., Cold Spring Harbor Press, pp. 811-835, 1977. Kouri, R.E., Rude, T., Thomas, P.E., and Whitmire, C.E. Studies on Pulmonary Aryl Hydrocarbon Hydroxylase Activity in Inbred Strains of Mice. Chem. Biol. Interactions, 13:317-331, 1976. Lamb, D. and L. Reid. Mitotic Rates, Goblet Cell Increase and Histochemical Changes in Mucus in Mice Bronchial Epithelium During Exposure to Sulphur Dioxide. J. Path. Bact. 96:97-111, 1968 Mitruka, B.M. and Rawnsley, H.M. Clinical Biochemical and Hematological Reference Values in Normal Experimental Animals. Second Edition. New York: Masson Publishing USA, Inc., 1981. Sokal, R.R. and Rohlf, F.J. Biometry, The Principles and Practice of Statistics in Biological Research. San Francisco: W.H. Freeman and Co., 1969. Young, J.T. Histopathologic Examination of the Rat Nasal Cavity, Fund & Applied Tox., 1:309-312, 1981. 24

QUALITY ASSURANCE STATEMENT Study Title: SIX WEEK REPEATED SMOKE INHALATION IN MICE TO COMPARE HISTOPATHOLOGY AND SHORT-TERM ENDPOINTS USING TEST (D6 AND Dl0) AND REFERENCE CIGARETTES Study Number: 11729.005 Study Director: Raymond M. David, Ph.D. Initiation Date: 83/03/24 Review Completed Date: 85/10/11 This study has been divided into a series of phases. Using a random sampling approach, Quality Assurance monitors each of these phases over a series of studies. Procedures, documenta- tion, equipment, etc., are examined in order to assure.that the study is performed in accordance with the Good Laboratory Practice regulations and to assure that the study is conducted according to the protocol. The following are the inspection dates, phases inspected, and report dates of QA inspections of the study. INSPECT ON 83/03/22 - 83/03/22,TO STUDY DIR 83/03/22,TO MGMT 83/03/22 PHASES: PROTOCOL REVIEW INSPECT ON 83/04/13 - 83/04/13,TO STUDY DIR 83/04/14,TO MGMT 83/07/01 PHASES: BLOOD COLLECTION FOR CLINICAL CHEMISTRY NECROPSY INSPECT ON 83/06/20 - 83/06/22,TO STUDY DIR 83/06/22,TO MGMT 83/07/01 PHASES: DRAFT REPORT INSPECT ON 83/06/30 - 83/07/01,TO STUDY DIR 83/07/01,TO MGMT 83/07/01 PHASES: PATHOLOGY REPORT INSPECT ON 85/10/07 - 85/10/07,TO STUDY DIR 85/10/07,TO MGMT 85/10/11 PHASES: DRAFT TO FINAL REPORT This report describes the methods and procedures used in the study and the reported results accurately reflect the raw data of the study. 1\ Quality Ass rance Date RA/QA Department 25

Final Report 1-1729.005 PATHOLOGY REPORT SIX WEEK REPEATED SMOKE INHALATION IN MICE TO COMPARE HISTOPATHOLOGY AND SHORT-TERM ENDPOINTS USING TEST (D6 and D10) AND REFERENCE CIGARETTES, STUDY NO. 1-1729.005 (CAHS No. 1263-1) INTRODUCTION AND METHODS Female B6C3F1/crlBL mice were divided into sham controls, a reference cigarette group, and 2 test cigarette groups and exposed to repeated doses of smoke by nose-only inhalation 5 days per week for 3 or 6 weeks. Twenty-one days after initiation of treatment, 3 mice from each group were anesthetized by metofane and killed by exsanguination. Forty-two days after initiation, 3 mice from each group were given colchicine (1 mg/kg) and 4 hours later were anesthetized with metofane and killed by exsanguination. Three additional non-colchicine-treated mice from each group were also anesthetized and killed in an identical manner. All mice were necropsied. Lungs were removed and weighed to the nearest 0.1 mg. The remaining organs and tissues were removed from the animal and fixed in 10% neutral buffered formalin. Lungs were fixed by perfusion with formalin to approximate the normal expanded state. The nasal passages were flushed with formalin and immersed in the fixative. Nasal passages were decalcified and trimmed according to the procedures described by Young (1981). Lung lobes were sectioned and embedded longitudinally through the stem bronchus of each lobe (Dungworth, et al, 1976); trachea and mainstem bronchi were removed and embedded whole longitudinally and 3 sections of the larynx were embedded transversely. All tissues except nasal passages were embedded in glycol methacrylate, sectioned at 2.5 um and stained with toluidine blue. The latter were embedded in paraffin, sectioned at 6 um, and stained with hematoxylin, phloxine, and eosin. Sections were evaluated microscopically without knowledge of the group being evaluated. RESULTS GROSS NECROPSY FINDINGS At both the 3-week interim and 6 week terminal sacrifices, treatment related gross lesions consisted of tan or brown discoloration of lungs of most smoke-exposed mice but not sham controls. No difference was discerned between reference and test cigarette groups. Other lesions were incidental (Table 1). 1

Final Report 1-1729.005 MICROSCOPIC FINDINGS Mice treated for 3 weeks had macrophage infiltration of pulmonary alveoli. It was most often observed in alveoli located near small airways. All mice of groups 1 and 2 and 1 of group 3 had this response. Lesions were graded as minimal to mild. Mice of group 4 (sham controls) were essentially free of this response. The response was interpreted to be attributable to smoke inhalation. A difference was not observed between reference and test cigarettes. All mice of both six weeks treatment groups (colchicine and non-colchicine), had minimal or mild alveolar macrophage infiltrates in the lung. The lesion was not observed in sham control lungs. Another response observed in both 3 and 6 weeks treated mice was lymphoid infiltration of the interstitium between small airways and vessels and infrequently, lymphoid infiltration around blood vessels of the lung. The response was minimal-to-mild in all instances. It was observed in 2 of the 9 smoke-exposed mice in the 3 weeks study and 17 of the 18 smoke-exposed mice in the 6 weeks studies. Similar lesions were not observed in sham controls. Nasal cavity, trachea and lung sections were assessed for changes in the airway epithelium. No morphologic changes were observed in test or control mice. DISCUSSION AND SUMMARY Smoke related responses were observed in the lungs of female mice exposed to smoke 5 days each week for 3 and 6 weeks respectively. The responses included macrophage infiltration of alveoli and lymphoid infiltration of the interstitial tissue between small airways and vessels and infrequently around pulmonary vessels. Grossly, lung discoloration was observed in most smoke-exposed mice, but not sham controls. Control mice were essentially free of gross and microscopic responses. No differences between reference smoke and test article smoke exposed groups could be discerned. 2

Final Report 1-1729.005 Signature Page &~ n/ 61301s3 Anthony son, D.V.M., Ph.D. Date Veterinary Pathologist Reviewed by: William C. Hall, V.M.D., Ph.D. Director of Veterinary Pathology Date 3

Final Report 1-1729.005 Leyend + = Tissue present NR = Organ not required 1 = Minimal 2 = Mild 3 = Moderate 4 = Severe TS = Terminal Sacrifice IS = Interim Sacrifice FD = Found Dead X = Lesion present but not graded - = Required organ not present

Final Report 1-1729.005 TABLE 1 Gross Necropsy Findings (CAHS 1263-1) GROUP INDIVIDUAL ANIMAL NO. GROSS OBSERVATIONS INTERIM SACRIFICE - 3 WEEKS 4 1187 Lungs - Red 1 1145 Lungs - Slightly tan 1 1147 2 1157 2 1158 2 1159 3 1169 3 1170 3 1170 Thymus - Slightly enlarged 3 1173 Gums - Pale 3 1173 Tongue - Pale 4 1187 Stomach - Distended with Air 4 1187 Upper Intestines - Distended with Air TERMINAL SACRIFICE - 6 WEEKS 1 1151 Lungs - Brown Discoloration 1 1152 1 1153 2 1163 2 1164 2 1165 3 1175 3 1176 3 1177 TERMINAL SACRIFICE - 6 WEEKS - COLCHICiNE TREATMENT 1160 Mesenteric Lymph Node - Hemorrhage 2 1161 Mesentery - Hemorrhage 2 1160 Lungs - Brown Discoloration T 2 1161 m 2 1162 t0 ~ 3 1171 T 3 1172 O 3 1174 ~ ~:

Final Report 1-1729.005 TABLE 2 Individual Animal Microscopic Findings (CAHS 1^63-1) (3 Weeks) Group Number• 1 1 1 2 2 2 3 3 3 4 4 4 An imal Number 1145 1146 1147 1157 1158 1159 1169 1170 1173 1181 1182 1187 Histology Number 83-1229 83-1230 83-1231 83-1232 83-1233 83-1234 83-1235 83-1236 83-1237 83-1238 83-1239 83-1240 Date of Death Type of Death 04/13/83 IS 04/13/83 04/13/83 04/13/83 04/13/83 04/13/83 04/13/83 IS IS IS IS IS IS 04/13/83 IS 04/04/83 FD 04/13/83 IS 04/13/83 IS 04/11/83 FD NASAL,ORAL,PHARYNGEAL SECTIONS + + + + + .1+ + + + + + LUNG + + + + + + + + + + + + Alveolar Macrophage 2 1 1 2 2 1 1 Accumulation Lymphoid Infiltration 1 2 Congestion LARYNX + - + + + + + + X + + - + TRACHEA + + + + + + + + + + + + THYMUS + 44094Egg

Final Report 1-1729.005 TABLE 2 (Cont'd) Individual Animal Microscopic Findings (CALIS 1263-1) (6 Weeks) Group Number 1 1 1 2 2 2 3 3 3 4 4 4 Animal Number 1151 1152 1153 1163 1164 1165 1175 1176 1177 1186 1188 1189 Ilistology Number 83-1244 83-1245 83-1246 83-1250 83-1251 83-1252 83-1256 83-1257 83-1258 83-1262 83-1263 83-12 64 Date of Death 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/0 4/83 05/04/83 Type of Death TS TS TS TS TS TS TS TS TS TS TS TS NASAL,ORAL,PHARYNGEAL SECTIONS + + + + + + + + + + + + LUNG + + + + + + + + + + + + Alveolar Macrophage 2 2 2 2 2 2 2 2 1 Accumulation L.ymphoid Infiltration 2 2, 1 1 1 1 2 2 LARYNX + + + + f + + + + + + + TRACIIEA + + + + + + + + + + + + Nf.lV J&GpR

Final Report 1-1729.005 TABLE 2(Cont'd) Individual Animal Microscopic Findings (CAHS 1263-1) - (6 Weeks - Colchicine Treatment) Group Number 1 1 1 2 2 2 3 3 3 4 4 4 Animal Number 1148 1149 1150 1160 1161 1162 1171 1172 1174 1183 1184 1185 Histology Number 83-1241 83-1242 83-1243 83-1247 83-1248 83-1249 83-1253 83-1254 83-1255 83-1259 83-1260 83-12 61 Date of Death 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/04/83 05/0 4/83 05/04/83 Type of Death TS TS TS TS TS TS TS TS TS TS TS TS NASAL, ORAL, PIIARYNGEAL SECTIONS + + + + + + + + + + + + LUNG + + + + + + + + + + + + Alveolar Macrophage 2 2 2 2 2 2 2 1 1 Accumulation Lymphoid Infiltration 2 2 2 2 1 1 2 1 1 LARYNX + + + + + + + + + + + + TRACHEA + + + + + + + + + + + i f4U9L6Bd