Document rBDxvYq7Nk16kamnZZ3bmRMO0

-^li-^'-f-*'***' EVALUATION OF THE ENVIRONMENTAL TOXICANTS VINYL CHLORIDE (VC) AND VINYLIDENE CHLORIDE (VDC) FINAL REPORT July 1975 through June 1977 Contract No. N01-ES-2-2084 (Continuation of NIH-NIEHS-72-2-20841 MRI Project No. 3612-8 ! ----------'--- ^- For National Institute of Environmental Health Sciences P.0. Box 12233 Research Triangle Park, NC 27709 Attn: Dr. James S. Woods SL 083811 i j MIDWEST RESEARCH INSTITUTE 425 VOIKER BOULEVARD, KANSAS CITY, MISSOURI 64110 816 753-7600 _ - O' i'diJ evaluation of the environmental toxicants VINYL CHLORIDE (VC) AND VINYLIDENE CHLORIDE (VDC) By Joseph M. Winston William B. House Paul J. Peters Jagdish C. 3handari John R. Hodgson Robert D. Short, Jr. Jack H. Hagensen Thomas W. Reddig Judith D. Girvin Ellen R. Ellis Cheng-Chun Lee FINAL REPORT July 1975 through June 1977 Contract No. N01-ES-2-2084 (Continuation of NTH-NIEHS-72-2-2084). MRI Project No. 3612-B \ National Institute of Environmental Health. Sciences P.0. Box 12233 Research Triangle Park, NC 27709. Attn: Dr. James S, Woods MID'VSST RESEARCH INSTITUTE SL 083812 -xenu ...^auii i, > n / J-2S VOLKER BOULEVARD, KANSAS CITY. MISSOURI 6*1 110 316 753-7SCQ PREFACE This report was prepared at Midwest Research Institute, 425 Volker Boulevard, Kansas City, Missouri 64110, under Contract No. N01-ES-2-2Q84 (Continuation of NIH-NIEHS-72-2084) with the National Institutes of Health, Department of Health, Education and Welfare, MRI Project No. 3612-B, "Research Capability for Environmental Health Sciences, Research Triangle Park, North Carolina 27709. Dr. James S. Woods is the Project Officer. This work was conducted in the Biological Sciences Division, under the direction of Dr. William B. House, between July 1975 and June 1977. Dr. Cheng-Chun Lee, Assistant Director, Biological Sciences for Pharmacology and Toxicology, was the Principal Investigator, assisted by Dr. Joseph M. Winston, Associate Toxicologist. Dr. William B. House supervised the inhala tion chamber operation, assisted by Dr. Paul J. Peters, Associate Biologist. Necropsy, gross and microscopic examination of tissues were performed by Dr. Jagdish C. Bhandari, Senior Pathologist, with the technical assistance of Mr. Ernesto Castillo and Miss Judith Shifrin. Dr. John R, Hodgson, Head, Biochemical and Development Pharmacology, supervised the cytogenetic analysis and collagen assays with the technical assistance of Mrs. Mary A. Kowalski and Mr. Don Van Goethem. Dr. Robert D. Short, Jr., Senior Toxicologist, per formed the thymidine incorporation studies and heme synthesis studies with the technical assistance of Miss Mary D. Sawyer. Mr. Jack R. Hagensen and Mrs. Karen J. Smith performed the analytical monitoring of chamber con centrations. Mr. Thomas W. Reddig, M.T. (ASCP certified) and Miss Judith D. Girvin, M.T. (ASCP certified) supervised the hematology and clinical labor atory tests with Che technical assistance of Miss Ilonna S, Elwood, Mr. Duane R. Smith and Mrs. Bhanu S. Gosalia. Mrs. Ellen R, Ellis, Histologist, super vised the histology preparation with the technical assistance of Mrs. Janet L. Klietthermes. Approved for: MIDWEST RESEARCH INSTITUTE Wra, abuse. Director Biological Sciences Division ill SL 083813 TABLE OF CONTENTS paae Abstract........................................................................................................................ I. Introduction............................................................................................ 1 II. Materials andMethods.......................................................................... 2 A. Methodsof Exposure............................................................... B. Safety.................................................................................... C. Animals.................................................................................... D. Experimental Protocol........................................................ 2 4 5 6 III. Results.................................................................................................. 11 A. Rats............................................................................................. 11 B. Mice............................................................................................. 15 C. Special Studies......................................................................... 20 IV. Discussion andConclusions....................................................................... 23 A. Rats.............................................................................................23 B. Mice............................................................................................. 24 C. Special Studies......................................................................... 27 References.................................................................................................................... 29 Tables 1-86 ............................................................................................................ 32-123 Figures 1-6 ............................................................................................................ 124-129 Appendix I - Hematology, Clinical Blood Chemistry, Urinalysis, Histopathology, Statistical Analysis, and Normal Values v SL 083814 EVALUATION 07 THE ENVIRONMENTAL TOXICANTS VINYL CHLORIDE (VC) AND VINYLIDENE CHLORIDE (VDC) Final Report ABSTRACT This report summarizes the results of up to 12 months exosure to various levels of vinyl chloride (VC) and one level of vinylldene chloride (VDC), 6 hr/day, 5 day3/week, in rats and mice. Exposure of rats of both sexes to 50, 250 or 1,000 ppm VC was without adverse effects through the first 7 months of exposure. Beginning with the 8th month, a number of rats died or were terminated before schedule. Hemangiosarcomas occurred in the liver and lung. Hepatic hemangiosarccmas were found In two males and 10 females exposed to 250 ppm VC and in six males and 15 females exposed to 1,000 ppm VC. Most of the rats with hemangiosarcomas In the liver also had hemangiosarcomas in the lung. Hemangiosarcomas of the lung were found in three fe males exposed to 250 ppm VC and four males and nine females exposued to 1,Q0Ql ppm VC. The occurrence of hemangiosarcomas in the liver and lung was both dose and time related. Additional hemangiosarcomas were found in other tis sues including skin, omentum, mesentery, and mesenteric lymph nodes. Exposure of rats to 55 ppm VDC produced few adverse effects. The weight gain of both male and female rats was less than controls during the final 3 months of exposure. Hemangiosarcomas occurred in the mesenteric lymph nodes of one male and in the skin of another male. In addition, lesions Including fatty changes and nodular or disseminated vacuolization in the liver were found in VDC-exposed rats. Exposure of mice of both sexes to 5Q, 250. or 1,000 ppm VC resulted in a large number of deaths and unscheduled terminations beginning with the 6th month of exposure. At the end of 12 months, only a few- mice exposed to 50 ppm VC survived. Bronchiolo-alveolar adenomas, first appeared during the second month, occurred in eight males and four females exposed to 250 ppm, and 22 males and 26 females exposed to 1,000 ppm. Hepatic--hemangiosarcomas first appeared during the 6th month, occurred in three males exposed to 50 ppm, seven males and 16 females exposed to 250 ppm, and 13 males and 18 females exposed to 1,000 ppm. Hemangiosarcomas also occurred in other tissues in cluding mammary gland, heart, skeletal muscle, gastrointestinal tract, kidney, pancreas, mesenteric lymph nodes, epidldymus, testes, and mesentery. The occurrence of the hemangiosarcomas was dose and time related. Mammary gland tumors consisting of ductular adenocarcinoma, squamous and/or anaplastic cell carcinoma with metastasis to the lung, also first appeared during the. 6th. month, occurred in female mice exposed to all levels of VC, The incidence of these tumors was greatest in the mice exposed to the highest level and for the longest period of time. Malignant lymphomas involving various organs were found in a few mice exposed to all levels of VC. vii SL 083815 A few tumors including bronchiolo-alevolar adenomas, hepatic hemangiosarcomas and hepatomas were found in mice exposed to VDC. Hepatic-hemangiosarcomas occurred in two males and one female, the relationship of the bronchiolo-aleveolar adenomas and hepatomas to the VDC exposure is question able. In addition, there were a number of lesions, including microfoci of mononuclear cells, focal degeneration and necrosis, intranuclear inclusions and/or large, basophilic nuclei, in the liver. Exposure of mice to 1,000 ppm VC or 55 ppm VDC also caused some acute deaths between the 3rd and 13th days. There was toxic hepatitis and marked tubular necrosis of the renal cortex. viii I. INTRODUCTION Under Contract No. N01-ES-2-2084 (NIH-NIEHS-72-2084) on "Research Capability for Environmental Toxicants," we have performed various studies on the toxic effects of the two commercially available dithiocarbamates, ferbam and thiram.i/ Following completion of these studies, the inhalation toxicity of various levels of vinyl chloride (VC) was chosen for study. In addition, one level of vinylidene chloride (VDC) was also included. Vinyl chloride was first prepared more than a century ago and its fire and explosion hazard are well known. Acute toxicity of VC was first reported in guinea pigs.1/ As a possible anesthetic agent in dogs, VC was found to cause incoordinated muscular activity of the extremities, cardiac arrhythmias^/ and sensitization of the myocardium.k! Relatively high con centrations of VC (20-40% in air) for 30 min produced narcosis and/or death in mice, rats, and guinea pigs..5/ The guinea pigs were found to be more resistant. The main lesions were congestion of the lung, with pulmonary edema and hemorrhages, in some animals, and congestion of the liver and kidneys. Blood coagulation was also Impaired. Various laboratory animals were exposed to 50, 100, 200 or 500 ppm of VC, 7 hr/day and 5 days/week, for up to 6 months../ A threshold limit value of 100 ppm, below which no detectable changes occurred, was suggested. A recent study indicated that rats (Ar/IRE) exposed to high concentration of VC (3% V/V in air, equal to 30,000 ppm), 4 hr/day, 5 days/week for 12 months, developed tumors of the skin, lungs, and,bones.U Furthermore, zymbal gland carcinomas, nephro blastoma, hepatic and extrahepatlc angiosarcomas were observed in rats, and pulmonary tumors, mammary carcinomas and liver angiosarcomas were observed in mice exposed to as low as 250 ppm of VC, 4 hr/day and 5 days^ week for 12 months.,/ The growing suspicion that exposure to VC has caused a number of deaths of workers from a rare form of liver cancer, angiosarcoma, has alarmed the OSHA to set an emergency rule on April 5, 1974, that lowered the permissible worker exposure level to 50 ppm from the previous level of 500 ppm. This report summarizes the final results of up to 12 months ex posure to VC or VDC. 083816 1 SL II. MATERIAL AND METHODS A. Methods of Exposure 1. Chamber design: Five stainless steel cubical type exposure chambers of 3.5 m-* (123 ft3) volume were used (Figure 1). Contaminant entered the air stream and was then mixed in a plenum at the top of each chamber. Each chamber contained a diffusion plate and two small squirrelcage fans (100 cfm, 2.85 m^/min) mounted on opposite sides of the top cone above the diffusion plate to ensure complete mixture of the gas with air. 2. Chamber air suonlv and flow rates: The air supply to the chambers was drawn from a stack on the roof of the building. Chamber air passed through a coarse filter and then over coils for heating, cooling, and dehumidifying. The air then passed through an absolute filter (99.9799.99% retention of 0.3 m particles) into the plenum of the chamber. Initially, air flow rates were measured at the inlet side of the chamber with a pitot tube connected to a magnahelix gauge. Subsequently, critical orifice plates were installed on the chamber outlet exhaust syst^ Air flow rates were then measured with an air flow transducer (Autotronics 100-SSX). The readings obtained with the critical orifice plates were greater than those obtained with the pitot tube. The correct air flow rate is 26.4 ft^/min (0.75 m^/min) or approximately 13 air changes per hour. 3. Generation of VC and VDC vapor: Vinyl chloride at a purity of 99.8% was obtained from Matheson Products. It is a gas at room tempera ture and was metered with rotameters into the chamber air supply. VDC with a purity of 99% was obtained from the Aldrich Company. Its boiling point is 32C (89.6F) and was heated to 37^ (98.6F) to generate the gas. All gas lines and the rotameter were heated to 40C (104F) to prevent conden sation. 4. Chamber atmosphere monitoring: Chamber concentrations were monitored using a gas chromatograph (Varian-2700) with a flame ionization detector. A 6 ft x 1/8 in. stainless steel column packed with 0.4% Carbowax 1500 on Carbopak A was used with a nitrogen carrier flow rate of 80 ml/min. The injection, column, and detector temperatures were 135C, 65C and 170C, respectively.5 5. Standards: VC standards at dilutions of 10, 50 and 100 ppm were obtained in lecture bottles from Supelco, Inc,, Bellefonte, Pennsylvania. VC standard gas at a concentration of 1,000 ppm was obtained from Matheson Gas Products, Joliet, Illinois. 2 SL 083817 VDC standards ware prepared by a serial dilution (weight/volume) of VDC in carbon tetrachloride. The desired final concentrations in a 4 ul injection were determined by the following procedure: mg/i (Mg/ml) * ppm x mw 24,450 With a molecular weight for VDC of 96,94, the final concentrations of 10, 50 and 100 ppm, the amount of VDC in a 1-ml sample from the chamber would be 0.04, 0.20 or 0.40 Mg respectively. For 1 m1 of standard, these same amounts would be obtained from solutions of 40, 200 and 400 mg/liter; but since 4 jil of standard were injected, the final dilutions of VDC in carbon tetrachloride were 10, 50 and 100 mg/liter. All liquids were injected at a volume of 4 m1 and all gases at 1 ml. Standards at each point on the calibration curve were injected in triplicate. 6. Chamber sampling: Each chamber was fitted with 10 sampling ports on two sides of the chamber. During preliminary studies, a series of distribution studies were made to determine the uniformity of the test material in the chamber. These tests were made with three sampling lines of polyethylene capillary tubing, of varying lengths, passed through each sampling port. It was determined that a valid measure of chamber contami nant distribution could be obtained by sampling periodically through three positions and occasionally through nine positions (Figure 2), Samples were withdrawn from the chamber with the same syringe used to introduce the sample to the gas chromatograph. Tests indicated that a valid sample could be with drawn by pumping the syringe three times on the short sampling lines, and five times on the longest line. All sampling was done in triplicate. During the first 3 months, distribution studies were compared with a reference point in the center of the chamber. The results showed that average chamber concentrations were 3Z of the desired concentration; the reference point averages were 98.4S to 100.4Z of these chamber averages. At a later point in the study, the manual sampling system was replaced by an automatic sampling system. The sampling system consisted of 1/4 in. teflon lines that were placed at the reference point in the chambers (see Figure 2). The teflon lines, which were purged with chamber atmosphere continuously, then ran through a series of sampling valves. Samples were Injected through twin 1-ml sampling loops which were allowed to reach equilibrium before the sample was injected. The sampling sequence was controlled by a Varian CDS-11T chromatography data system. The concentrations were determined by integration of peak areas by the CDS-111'5'. The CDS-111 was calibrated each morning with VC and VDC standards. 3 SL 083818 The average weekly concentrations and the range of all samples for VC are shown in Figure 3. The weekly variations for the average chamber concentration of VC did not vary more than t 57, from the desired values. With a few exceptions, the range of individual samples did not vary more than 10% from the desired concentration. At the beginning of testing with VDC, it was planned that the exposure level would be 50 ppm. Due to technical problems with the standards, the objective of 50 ppm was not achieved. These problems were associated with the use of chloroform as the solvent during the first 4 weeks, which was subsequently changed to carbon tetrachloride, and a nonlinear calibra tion curve during the 5th through the 10th week. By the 11th week, these problems had been solved and the slightly higher exposure level of 55 ppm was obtained. The average weekly concentrations of VDC are shown in Figure 4, as well as the range of individual samples. Due to the problems discussed above, the analytical concentration during the first 10 weeks is not pre cisely known. Since air flow rates have been constant throughout, we can calculate the nominal concentration using the amount of VDC used each day. This has been done (Figure 4) for the 4th through the 10th week. However, the material balance was not recorded during the first 3 weeks. Therefore, we have estimated the average concentration during the first 3 weeks by replicating all magnahelic and rotameter settings, which were known, and measuring the chamber concentration. By this method, we estimate the average concentration during the first 3 weeks to be 30, 80 and 95 ppm. The average chamber concentrations from the 11th week on were generally within t 5% of the desired 55 ppm VDC. Exceptions to this were weeks 27, 28, 44 and 45. The range of individual samples was within 15% of the desired 55 ppm VDC. B. Safety All chambers were operated with a slight negative pressure (0.10.2 in. water) to prevent escape of contaminant through leaks into the room. Before opening, chambers were flushed with clean air for a minimum of 30 min. Sampling from the chambers and from the doorway upon opening always showed less than 1 ppm of contaminant. In addition, after operating for 30 min without contaminant, on one occasion, the air supply and exhaust were shut off to the 1,000 ppm chamber for 10 min. Thus, the animals sat in a static environment and any off-gassing would be allowed to accumulate. Samples taken from the chamber after the 10-min period showed less than 1 ppm of VC in the air. 4 SL 083819 Additional samples from doorways and numerous other places in the area were collected on charcoal tubes and by grab sampling with a syringe. All samples showed less than 1 ppm of VC. Room monitoring for VC and VDC was performed on a routine basis by grab samples with a syringe and analysis with the gas chromatograph used for chamber monitoring. During all initial work, personnel wore impervious coveralls, gloves, and a demand type face mask when the chambers were opened for trans fer of animals to holding racks. This practice was discontinued only after extensive sampling had shown that exposure to more than 1 ppm did not occur. All air from the chambers travels under negative pressure to an incinerator outside the building. Integrated bag samples of stack gas were collected on January 12 and 13, 1976. These samples were collected accord ing to Method 106, ''Determination of Vinyl Chloride from Stationary Sources," Federal Register, Vol. 40, No. 243, December 24, 1975. This method involves pulling a sample of stack gas into a leak proof Tedlar bag with a vacuum pump set at a flow rate of 500 cc/min for a 1-hr period. The sample site was de termined by cutting a hole in the stack wall at two duct diameters downstream from the flame. Two tests were conducted on January 12, while three chambers operated at a combined level of 1,300 ppm of VC; and one chamber was operated at 55 ppm of VDC. An additional three chambers operated with only clean air. Test one, analyzed at the inhalation facility, showed 39 to 40 ppm of VC. Test two showed 33.5 to 39 ppm of VC. To check these values, a sample was drawn by syringe directly from the stack. This sample contained 29 ppm of VC. VDC was never detected. On January 13, a third test was conducted and showed 38 ppm of VC. Samples from the third test were taken to the Environmental Measurements Section at MRI. After setting overnight, the average of 6 peaks equalled 29 ppm of VC. This lower value could be due to loss of VC through absorp tion in the Tedlar bag, or due to loss through the used septum. During all three tests, the burner of the incinerator operated at 1700 to 1800?. Subsequent tests at higher or lower temperatures did not lower the VC. C. Animals 1. Species and Albino CD rats and albino Swiss mice (Charles River Breeding Laboratory), about 2 months old at the start, were used in these studies. They were acclimated in our holding quarters for 1 week before the experiment began. For each species, a total of 360 animals were divided into five groups, each consisting of 36 males and 36 females. 5 SL 083820 2. Animal housing: All animals lived in the same stainless steel cages during exposure and outside of the chambers. Rats were housed " two per cage and mice six to eight per cage. Powdered or block laboratory chow (Wayne Manufacturing Company) was provided at all times except during the 6-hr period of exposure each day. Water was available ad libitum. The temperature in the chamber and in the room averaged 24 1.3C (75 5F). The relative humidity in the chambers ranged from 25 to 60%; in the room it also ranged from 25 to 60%, but was regulated at 45 i 5% after the 6th month. A 12-hr light cycle was maintained at all times. When out of the chambers, the animals were placed in holding racks similar to those in Figure 1. A continuous supply of room air was drawn over the cages while in the holding racks. Provisions were made for covering the front of each compartment in the holding rack in the event of significant off-gassing from the animals. This was found not to be necessary with VC or VDC. 3. Animal hygiene: All cages and feeders were washed and sani tized each week. The floors of the room, the holding racks, and the in terior of the chambers were frequently washed with disinfectant. Animal caretakers wore gloves and surgical masks when handling animals. It was found that when the cages were loaded into the chambers without drop-pans (so not to impair air flow) the animals on the bottom shelves were frequently drenched with urine and water. In order to allev iate this problem, the cages were loaded into the chambers with clean drop pans in place. Sampling studies showed that the vapor distribution was actually improved slightly. Thus, from the 6th month on, drop pans were washed daily and a clean drop pan without bedding was inserted into each cage before loading into the chamber. D. Experimental Protocol 1. Experimental design: A total of five chambers were used. Each chamber held one group of 72 rats and one group of 72 mice. Animals in the chambers were exposed at 6 hr/day and 5 days/week to the following: Chamber 1: Chamber 2: Chamber 3: Chamber 4: Chamber 5: Uncontaminated air 50 ppm of VC 250 ppm of VC 1,000 ppm of VC 55 ppm of VDC 6 SL 083821 Four animals of each species, sex and exposure level, were termi nated for various studies, necropsy, and histopathology at the end of 1, 2, 3, 6 and 9 months. At the conclusion of 12 months exposure, all remaining animals were terminated. Moribund animals were terminated as required. All terminated animals were replaced in the respective chambers at periodic in tervals. The replacement animals are exposed for 1, 3 and 6 months for mice and 1, 3, 6 and 10 months for rats. At the conclusion of the exposure periods, the animals are removed from the chambers and held for 1 year for observation. At the end of 1 year, all animals will be terminated for necropsy and histo pathology. 2. Observations and studies: a. General observations and weight gain: All animals were observed throughout the study for behavioral changes and any unusual or toxic signs. Food consumption was recorded weekly and body weights biweekly at a uniform time of the day. Any unusual changes in weight were investi gated. An attempt was made to determine the cause when any unscheduled death occurred. b. Hematology and clinical blood chemistry; Aortic blood from males and females of each group of each species was collected under anesthesia (sodium pentobarbital for rats and ether for mice) at each termination period. Hematology (various blood counts, hematocrit, hemo globin, methemeglobin and Heinz bodies) and clinical blood chemistry (fasting blood glucose, SGOT, SGFT, alkaline phosphatase, BUN, creatinine and/or LDH) were performed on all samples. The procedures for these tests and the normal values for rats are given in Appendix I. In addition, the following tests were also performed for rats. (1) Prothrombin time: The prothrombin time was measured by the one-step method of QuickC/ using the Fibro System (BBL) with activated thromboplastin (Dade). Standardized Normal Plasma (Dade) was used as the control for each assay. (2) Bilirubin: Total and direct bilirubin was measured using the Billi-Strate Kit (General Diagnostics) which is based on a modified Jendrassik-Grof method.i2/ Moni-Trol I, Versatol (General Diagnostics), and Calibrate I (General Diagnostics) was used as the reference for each assay. c. Serum -tmratmogiobins: Serum immunoglobins were measured in four male and four female rats from the control, 1,000 ppm of VC, or 55 ppm of VDC groups. The radial immunodiffusion technique of Mancini et al.^ was used to assay immunoglobin groups IgA, IgG, subgroups A and B, and IgM. 7 SL 083822 A replicate serum sample from each rat was placed la wells in an immunodiffusion chamber along with a standard, and held until the precipitin ring reached equilibrium. The square root of the diameter of the precipitan ring is directly proportional to the concentration of anti body. d. Serum protein: (1) Total protein: Total protein was measured by the biuret method of Kingsley3Z7 Standard Human Protein (Dade) was used as the standard for each assay. (2) Albumin: Albumin was measured by the Albu-Strate Kit (General Diagnostics) which is based on the bromcresol green method of Doumas et al.,H' Lab-Trol (Dade) and Calibrate I was used as the reference for each assay. (3) Globulin: Globulin was calculated by the difference of total protein and albumin. e. Pulmonary macrophages: Pour males and four females of each species from the control, 1,000 ppm of VC, or 55 ppm of VDC groups were used for the pulmonary macrophage count. At the time of necropsy, the left lung was removed at the bifurcation of the trachea; a small needle was inserted into the bronchus; and 5 ml- of the sterile saline was gently flushed into the lungs of rats and 0.5 ml in the lungs of mice. The fluid was withdrawn and flushed again several times. This fluid was then transferred from the syringe to a glass tube and stained. The number of cells in the preparation was counted under a microscope with a differentiation made between macrophages and leukocytes. If changes were observed in lungs from animals exposed to 1,000 ppm of VC, then animals from the groups exposed to 250 or 50 ppm were also examined. f. Cytogenetic analysis: Bone marrow samples were collected from four males and four females of each species from the control, 1,000 ppm of VC, or 55 ppm of VDC groups. Short-term bone marrow cultures were made according to the methods of Tjlo and Wang.--/ After allowing approximately 5 hr for sufficient members of cells to be arrested in metaphase, the cells were collected, swollen in hypotonic saline, and processed for spreading on glass slides according to the method of Moorhead and Norwell.H/ Slides were then stained with giesma and scanned under low power optics. Those slides showing a m-tn-tTmim scattering of cells in metaphase were selected for chromosomal analysis using oil immersion optics. Estimation of cell polyploidy was obtained by rough chromosomal estimates of at least 200 cells. Exact chromosomal counts and chromosomal aberration estimates were determined for at least 50 metaphase spreads. Special attention was given to chromosomal damage such as breaks, gaps, and translocations. 8 SL 083823 g. Eyam-fnation of bones: At the time of scheduled necrop sies, the hind limbs from both species were excised and fixed in formalin. After fixation, the limbs were wrapped in plastic and frozen for storage. Following the 12 months terminations, limbs were examined for evidence of osteolysis or ocher changes. The limbs were examined using a senograph X-ray machine set at 10 mA, 0.4 sec and 22 KVF with a large mammographic cone and focal film distance of 37 cm. The specimens were radiographed with Kodak MHT-R film. The radiographs were then viewed as unknowns and evaluated for any radio graphically discemable differences between groups. They were examined for the presence of any bone tumors, any changes in bone density, cortical thick ness or striations within the bone cortex, any loss of bone cortex in terms of total width and/or subperiosteal or endosteal erosion, or any unusually widened, narrowed or distorted trabecular pattern of the bone. h. Necropsy and histopathology: When moribund or at the end of each termination period, males and females from all groups were euthanized for necropsy after the collection of blood. Gross examination of all tissues, especially for any appearance of abnormal growth or other lesions, were carefully performed. The brain, liver, kidney, spleen and gonads were removed and weighed. Portions of these and other tissues were fixed, processed, sectioned and stained for microscopic examination. The procedures are given in Appendix I. 3. Statistical analysis: The results of the various parameters were compared with the control groups at the respective time intervals, whenever applicable, according to the Dunnett's Multiple Comparison Pro cedures ^ 4. Special studies: a. Thymidine incorporation into DNA: Male mice were ex posed to SO ppm VC for 43 to 45 weeks as previously described. On the day following the last exposure, mice were injected with 100 MCl/kg of thymidine ^2"^C from New England Nuclear (Boston, Massachusetts) . One hour following the injection of ^C-thymidine, mice were sacrificed by cervical dislocation and the liver was removed and weighed. A portion of the liver (2-300 mg) was saved for histopathological examination while the remaining portion (>1.3 g) was homogenized in 5 ml of cold water. Macro-molecules were precipitated by the addition of 5 ml of 1 N perchloric acid (PCA). The resulting precipitate was washed with successive 5 ml por tions of 0.2 N PCA, 0.2 N PCA, 95% ethanol saturated with sodium acetate, ethanol:ether (3:1). The precipitate was dissolved in 4 ml of 0.3 N sodium hydroxide and heated for 1 hr at 37C. The mixture was acidified by the addition of 1.8 ml of 50% trichloroacetic acid (TCA). The precipitate was washed twice with 5 ml of 5% TCA and heated at 9038C4 f*o*r* 20 min in 5 ml of 9 SL 083824 5% TCA. The supernatant was saved and the precipitate was heated again in 52 TCA. This supernatant was saved and combined with the previous super- ' natant. Aliquots of the hot acid extract were used for liquid scintillation counting and DNA determination.HJ The results were expressed as DFM/mg DNA and mg DNA/g liver. b. 3-Amlnolevullnic acid svnthetase (ALAS) and urinary jMi~inolevulinic acid (ALA) assays*. Rats were exposed to various concentrations of VC for 6 hr a day, 5 days a week, for 12 months. The ALAS assay described by Wood&i^ was used in this study. Basically the method involves the prepara tion of a mitochondrial fraction from homogenates of rat liver and the incuba tion of 0.5 ml of this fraction with 50 mM Tris-HCl buffer (pH-8.5), 10 mM MgCl2, 100 mM glycine, 10 mM sodium succinate, 0.2 mM pyridoxal phosphate, 1 mM (3-mercaptoethanol, 250 mM sodium chloride, 0.1 mM GTP, 5 mM EDTA, 60 pM coenzyme A, 3 mM ATP and 0.2 units of succinyl thiokinase (one unit converts 1 mole of succinate to succinyl-Co A per minute at pH 7.4 at 30 C) for a final volume of 2.5 ml. All reactions were conducted at 37flC for 1 hr in room air. Reactions were terminated with 0.5 ml of 102 TCA. ALA was con verted to the pyrole derivative and quantified with modified Ehrlich's Reagent using a molar extinction coefficient of 5.3 x 10^ as previously described.--' Proteins in the mitochondrial fraction were quantified by the method of Lowry et al.,12./ The Bio-Rad Urinary ALA Test Units (Bio-Rad Laboratories, Richmond, California) were used in this study. The assay is based on the method described by Davis.--/ Basically, the method involves separating ALA from porphobilinogen on ion-exchange columns, eluting the ALA from the column, and producing a pyrole derivative of ALA which is detected with Ehrlich's Reagent. Urine was collected overnight from rats housed in metabolism cages with free access to feed and water. c. Alpha-fetoprotein assay: Serum samples from selected rats and mice were retained for assay of alpha-fetoprotein levels.21/ The assays of alpha-fetoprotein were performed by Dr. Stewart Sell, Department of Pathology, School of Medicine, University of California at San Diego, LaJolla, California. d. Dominant lethal study: Groups of male rats that were in the 11th and 12th week of exposure to VC or VDC were used in this study. In the evenings following exposure, each male was housed overnight with two nonexposed virgin female CDrats. This procedure was followed for seven successive evenings or until a male mated with two females. All females were examined in the morning for evidence of mating. On gesta tional day 13, females were sacrificed and the number of corpora lutea and implants (viable and dead) determined. 10 SL 083825 III. RESULTS A. Rats 1. General observations and weight gain: Rats were exposed to VC or VDC, 6 hr/day, 5 days/week, for periods of 1, 2, 3, 6, 9 or 12 months. During these periods, these rats were exposed for a total of 22, 41, 61, 131, 196 or 259 days, respectively. All groups of rats gained weight and appeared to be in good health until the final 1 to 2 months of exposure. The body weights of the control rats and rat3 exposed to 1,000 ppm VC or 55 ppm VDC are shown in Figure 5. During most of the exposure, the body weights of the control rats and rats exposed to VC or VDC were comparable. However, the body weights of the males and females exposed to 1,000 ppm VC decreased as compared to controls after 43 weeks of exposure in the females and after 49 weeks of exposure in the males. The body weights of the males and females exposed to 55 ppm VDC were less than those of controls during the final 3 months of exposure. Although not shown in Figure 5, the body weights of the male and female rats exposed to 50 or 250 ppm VC were comparable to controls. The final 1 to 2 months of exposure to VC was marked by a substantial number of terminations, due to ill health and deaths. These terminations and deaths occurred primarily in the 250 and 1,000 ppm VC exposed groups. 2. Laboratory data: The results of hematology, clinical blood chemistry, serum iamunoglobins, serum proteins, and macrophage counts in the lungs of male and female rats after exposure to VC or VDC for 1, 2, 3, 6, 9 or 12 months are summarized in Tables 1 through 12. At each of the periods of observation, the peripheral blood elements and clinical chemistry values were within normal ranges for rats observed in our laboratory as shown in Appendix I. When compared with control rats, there were a number of oc casional differences at each observation period. However, these differences do not appear to be related to the VC or VDC exposures. At the end of 1 month, in the male rats (Table 1) the SGOT level was lower and the alkaline phosphatase level was higher in the group ex posed to 50 ppm VC, and the direct bilirubin concentration was higher in the group exposed to 250 ppm VC or 55 ppm VDC. In the female rats (Table 2), the leukocyte counts, creatinine concentration and total protein were higher in the group exposed to 55 ppm VDC. At the end of 2 months, in the male rats (Table 3), the leukocyte count was higher in the group exposed to 50 ppm VC, the B factor of immunoglobln G was lower in the group exposed to 1,000 ppm VC, and the reticulo cyte count was higher and the hematocrit was lower in the group exposed to 55 ppm VDC. SGOT levels were lower in all groups exposed to VC as well as VDC. SGPT levels were lower in the groups exposed to 250 ppm VC or 55 ppm 11 SL 083826 VDC. The lactic dehydrogenase (LDH) and alpha-hydroxybutyrate dehydro genase (alpha-H3DH) levels were lower in all groups exposed to VC. In the female rats (Table 4), the percentage of reticulocytes was higher in the group exposed to 250 ppm VC. At the end of 3 months, the prothrombin time in male rats exposed to 1,000 ppm VC and immuneglobin A and the A factor of immunoglobin G in males exposed to 55 ppm VDC were lower than controls (Table 5). In the female rats (Table 6), the pulmonary macrophage counts of the group exposed to 1,000 ppm VC or 55 ppm VDC, the hemoglobin, mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCHb) indices, and the imniunoglobin A of the group exposed to 55 ppm VDC were lower than the respective controls. At the end of 6 months, the prothrombin time of the male rats exposed to 250 ppm VC and the serum immunoglobin M level of the males ex posed to 55 ppm VDC were lower, and the creatinine level of the males exposed to 1,000 ppm VC was higher than controls (Table 7). In the female rats (Table 8), the MCHbC index and the serum albumin level was higher in the group exposed to 50 ppm VC; the MCHb index was higher in the group exposed to 1,000 ppm VC, the hemoglobin and the immunoglobin M levels were lower and the immunoglobin A level was higher in the group exposed to 55 ppm VDC than the respective controls. At the end of 9 months, the LDH and a-HBDH levels of male rats exposed to all levels of VC were lower than controls (Table 9). Xn addi tion, the percentage of monocytes were lower and the SGOT level was higher in males exposed to 50 ppm VC or 55 ppm VDC; the erythrocyte count and serum immunoglobin M level were lower, and the MCV and MCHb indices were higher in males exposed to 55 ppm VDC than in controls. There were no significant changes in female rats exposed to VC or VDC for 9 months (Table 10). At the end of 12 months, the LDH and/or a-HBDH levels was lower in male rats exposed to 50 or 250 ppm VC (Table 11) The serum immunoglobin A and/or M was lower in males exposed to 1,000 ppm VC or 55 ppm VDC, In the female rats (Table 12), increases in percentage of reticulocytes, leuko cyte count, SGOT, SGPT, alkaline phosphatase levels and pulmonary macrophage count occurred in the group exposed to 1,000 ppm VC. However, the measure ments were performed in only two females and the increases were due to the extremely high values found in one of these rats (Ho. 275). 3. Cytogenetic analysis; Bone marrow cultures were prepared from both sexes of control rats and rats exposed to 1,000 ppm VC or 55 ppm VDC at each termination period. The results of these studies are presented in Tables 13 and 14. The bone marrow preparations from rats exposed to 1,000 ppm VC or 55 ppm VDC, for each of the exposure periods studied, showed no changes in chromosome frequency, distribution or number of tetTaploids, or any changes in the frequency of chromatid breaks, gaps or translocations. 12 SL 083827 4. Examination of bones; X-ray examination of the hind limbs of rats terminated after 12 months exposure to VC or VDC showed no evidence of osteolysis or other abnormalities. The bones from rats terminated at earlier periods were removed, fixed and/or frozen and stored, but not examined. 5. Collagen content in liver and lung: The collagen content in the liver and lungs of control rats and rats exposed to 1,000 ppm VC or 55 ppm VDC for 6, 9 or 12 months is summarized in Tables 15 to 17. No signifi cant changes in collagen content were found In the livers or lungs of rats exposed to 1,000 ppm VC or 55 ppm VDC for 6, 9 or 12 months. 6. Organ weights: The organ weights of male and female rats exposed to VC or VDC for 1, 2, 3, 6, 9 or 12 months are summarized in Tables 18 to 29. At 2 months, the absolute brain weight of female rats exposed to 55 ppm VDC was statistically smaller than that of the controls although this difference was small (Table 21). At 6 months, the absolute spleen weights of male rats exposed to 1,000 ppm VC or 55 ppm VDC were smaller than con trols (Table 24). Based on brain weight, however, the difference was not significant. The absolute brain weight of females exposed to 55 ppm VDC for 6 months was smaller than control values (Table 25). At 12 months, the absolute liver weight and liver weight relative to brain weight were both significantly higher in female rats exposed to 1,000 ppm VC (Table 29). However, there were only two rats in this group and the increase was due to one of the rats (No. 275) which had a hepatic hemangiosarcoma. With the exception of the changes just noted, there were no other differences in organ weights at any of the observation times. 7. Gross and microscopic examination of tissues: a. Gross changes: Gross examination of rats exposed to VC or VDC for 1, 2, 3 or 6 months revealed no remarkable lesions or tumors. Beginning about the end of the 3th month, a number of rats died or were terminated before scheduled because of 111 healths The number of deaths and early terminations increased sharply during the final 3 months of ex posure. Gross examination of rats exposed to VC terminated after 9 or 12 months or terminated before scheduled showed a number of lesions. The livers from these rats appeared mottled, with, spots ranging from pin-head size to several millimeters in diameter or had small white, tan or red nodules in the liver. Lungs obtained from numerous rats also had a red mottled appearance. In addition, some subcutaneous tumors ranging In size from 2 to 5.5 cm and greyish white to dark in color were noted. SL 083828 13 b. Tumors la racs exposed to VC or VDC: Microscopic exami nation of tissues showed a number of tumors. The tumor occurrence in rats exposed 7. to 9 months and 10 to 12 months are summarized in Tables 30 to 32. The cumulative incidence of all tumors is summarized in Table 33. The only tumor found in control rats was a ductular adenocarcinoma in the mammary gland in one female control. The most prevalent tumors found in rats exposed to VC were hepatic and pulmonary hemangiosarcoma. Hepatic hemangiosarcoma occurred in two females exposed to 250 ppm VC and four females exposed to 1,000 ppm VC during the 35th through 39th weeks (Table 30). An additional two males and eight females exposed to 250 ppm and six males and 11 females exposed to 1,000 ppm developed hemangiosarcoma during the 40th through 52nd weeks (Tables 31 and 32). Most c" these rats also developed hemangiosarcoma in the lung. Pulmonary hemani; 'arcoma occurred in one female exposed to 250 ppm and one female exposed : L,000 ppm during the 39th week (Table 30). An additional two females e:: sed to 250 ppm and four males and eight females exposed to 1,000 ppm develo^c pulmonary hemangiosarcoma during the 43rd through 52nd weeks (Tables . . and 32). No hemangiosarcoma was found in the liver or lung of rats expose to 50 ppm VC or 55 ppm VDC. Hemangiosarcoma occurred occasionally in other tissues also (Tables 30-32). These included one rat (subcutaneous) exposed to 50 ppm VC, two rats (omentum or mesentery) exposed to 250 ppm, five rats (omentum or mesentery) exposed to 1,000 ppm, and two rats (subcutaneous or mesenteric lymph nodes) exposed to 55 ppm VDC. Malignant lymphoma occurred in one female exposed to 250 ppm VC for 52 weeks, one male exposed to 1,000 ppm VC for 52 weeks and in two females exposed to 1,000 ppm VC for 47 to 49 weeks. The malignant lymphoma in the male (No. 242, Table 32) exposed to 1,000 ppm VC was widely disseminated being found in adrenal, heart, lung, liver, mesenteric lymph nodes, spleen, kidney, intestine, prostate and pancreas. The malignant lymphomas found in the other rats appeared to be confined to the spleen. Other tumors which occurred in one or several rats were bronchiolar adenoma and squamous cell carcinoma of the lung; reticuloendothelial cell carcinoma and hepatoma in the liver; ductular adenocarcinoma and fibroadenoma In the mammary gland; renal adenoma; hemangioma of the adrenal gland; squamous cell carcinoma, keratocanthoma or fibroma in the skin; adeno carcinoma in the sebaceous gland; and chromophobe cell adenoma in the pitui tary (Table 33). These occasional tumors were not related to VC or VDC. c. Other lesions: There were a number of spontaneous or incidental lesions in various tissues of control rats and rats exposed to VC or VDC for various lengths of time (Tables 34-39). These lesions were not related to VC or VDC. However, there were a few lesions in the livers of rats exposed to 55 ppm VDC. These lesions included fatty changes in the liver after exposure for 6 and 9 months (Tables 37 and 38) and nodular or disseminated vacuolization after exposure for 12 months (Table 39). Generally, these lesions were more severe and the incidence was greater than in control rats. 14 SL 083829 The myeloid/erythroid (M/E) ratios of the rib bone marrow smears of control rats and rats exposed to VC or VDC were within normal limits at all time periods examined (Tables 34-39). B. Mice 1. General observations and weight gain: The mice were exposed to VC or VDC, 6 hr/day, 5 days/week, for periods of 1, 2, 3, 6, 9 or 12 months. During these periods, these mice were exposed for a total of 19, 39, 59, 128, 193 or 256 days, respectively. There were five early deaths, apparently due to the acute effect of the test compounds. Two male and one female mice exposed to the highest level of VC were found dead between the 3rd and 9th days of exposure; two males exposed to VDC died on the 13th day. They were replaced with healthy mice from the same shipment for the remainder of the experiment. Thereafter, all mice appeared in good health. During the 6th month of exposure, the health of a few mice ex posed to various levels of VC deteriorated. The clinical signs included rough hair coat, lethargy, anorexia and rapid weight loss. They died or were terminated before their imminent death. During the 7th through the 9th months, the general health of the mice worsened. Additional clinical signs were abdominal distention and/or the appearance of external tumor masses, especially mammary tumors in the females. There were a large number of deaths or unscheduled terminations, proportional to exposure VC levels. By the end of the 9th month, all males and females in the group exposed to 1,000 ppm and all females exposed to 250 ppm died or were termi nated. By the end of 12 months, only the controls and a few mice exposed to 50 ppm VC had survived. In contrast, only a few deaths or early termi nations occurred in the VDC exposed group. The number of deaths and early terminations during the final 6 months of exposure is shown in Table 40. The average weight gains of the mice exposed to 1,000 ppm VC or 55 ppm VDC are shown in Figure 6. The weight gains of these mice were com parable to controls through the 35th week of exposure. The weight gain of the male mice exposed to 1,000 ppm VC began to decline at week 36 and re mained less than controls through week 39. The body weights of the females exposed to 1,000 ppm VC followed this same pattern although the weight loss during weeks 37 and 38 was exaggerated since only one or two animals remained in the group. The body weights of the mice exposed to 55 ppm VDC remained similar to those of controls although during the period of weeks 46 through 51 the weights were less than the controls. The body weights of the mice exposed to 50 and 250 ppm VC, although not shown in Figure 6, were similar to that of the controls throughout the exposure period. 15 SL 083830 2. Laboratory data: The results of hematology, clinical blood chemistry, and pulmonary macrophage counts in male and female mice exposed to VC or VDC for 1, 2, 3, 6, 9 or 12 months are summarized in Tables 41 to 52. At each of the observation periods, the peripheral blood elements and clinical blood chemistry values vere within normal ranges. When compared with control mice, there were some occasional differences at the respective observation period. However, these differences were small and not seen consistently. At the end of 1 month, the MCV index of male mice exposed to 50 ppm VC, the MCHb index of male mice exposed to 250 ppm VC, and the SGPT level of male mice exposed to 55 ppm VDC were higher than controls (Table 41). At the end of 2 months, the percentage of eosinophils of male mice exposed to 50 ppm VC, the reticulocyte count and BUN levels of female mice exposed to 55 -pm VDC were higher, and the BUN levels of male mice at all levels of VC -are lower than controls (Tables 43 and 44). At the end of 3 months, statistical difference was noc found between any values of the control and the exposed mice (Tables 45 and 46). At the end of 6 months, the male mice exposed to 1,000 ppm VC had lower hematocrit and hemoglobin concentration; the percentage of neutrophils of these mice was higher with a corresponding lower percentage of lymphocytes (Table 47). At the end of 6 months, the pulmonary macrophage counts of male and female mice exposed to 1,000 ppm VC, were higher than controls (Tables 47 and 48). At the end of 9 months, the pulmonary macrophage count of male mice exposed to 1,000 ppm VC was higher than controls (Table 49). In female mice exposed to 2 or 1,000 ppm VC (Table 50), there were several differences as compared tc the controls. However, only one animal was examined in each group and no statistical comparison was made. At the end of 12 months, the SGPT and BUN levels of male mice exposed to 55 ppm VDC were higher than controls (Table 51). In female mice exposed to 55 ppm VDC (Table 52), the percentage of reticulocytes and BUN level were higher than controls and the percentag of monocytes was lower than controls. 3. Cytogenetic analysis: Bone marrow cultures were prepared from both sexes of control mice and mice exposed to 1,000 ppm VC or 55 ppm VDC at each termination period. The results of these studies are summarized in Tables 53 and 54. The bone marrow preparations from mice exposed to 1,000 ppm VC or 55 ppm VDC showed no changes in chromosome frequency, distribution or number of tetraploids, or any changes in the frequency of chromatid breaks, gaps or translocations. 4. Exjtm-fnation of bones: X-ray examination of the hind limbs from mice exposed to VC or VDC for 12 months or from those terminated before schedule showed no evidence of osteolysis or other abnormalities. The hind limbs of mice exposed to VC or VDC for 9 months or less were re moved, fixed and/or frozen and stored, but not examined. 16 SL 083831 5. Organ weights: The organ weights of mice exposed to VC or VDC for 1, 2, 3, 6, 9 or 12 months are summarized in Tables 55 to 66. At 1 month,, the absolute brain weight in male mice exposed to all levels of VC and VDC and the absolute spleen weight in male mice exposed to 55 ppm VDC were significantly lower than controls (Table 55). The relative spleen to brain weight in female mice exposed to 250 ppm VC for 2 months (Table 58) and the relative spleen a-nri testes weight in male mice exposed to 50 ppm VC for 3 months (Table 59) were statistically larger than controls. At 9 months, the brain weights of male mice exposed to 50 ppm VC or 55 ppm VDC and the testes weights of male mice exposed to 50 or 1,000 ppm VC were smaller than the respective controls (Table 63). No significant changes in organ weights were seen in female mice at 9 months. However, the one female survivor at 9 months exposed to 250 ppm VC had a greatly enlarged liver and spleen (Table 64). Histopathologic examination showed that this mouse had a hepatic hemangiosarcoma and lymphoid hyperplasia of the spleen. At 12 months, the absolute kidney weight and the kidney weight relative to brain weight in male mice exposed to 55 ppm VDC were significantly smaller than controls (Table 65). In female mice exposed to 55 ppm VDC for 12 months, the absolute liver weight and liver and spleen weights relative to brain weight were signifi cantly larger than the respective controls (Table 66). No significant change was seen in any mice exposed to 50 ppm VC and no mice exposed to 250 or 1,000 ppm VC survived for 12 months. 6. Gross and microscopic examination of tissues: a. Early deaths: Microscopic examination revealed a number of lesions in the five unscheduled deaths (two males and one female exposed to 1,000 VC for 3 to 9 days, and two males exposed to 55 ppm VDC for 13 days). These lesions included acute toxic hepatitis, characterized by focal to marked congestion, and marked diffuse coagulation type necrosis of hepatocytes beginning in the centrilobular region of the liver. Marked tubular necrosis characterized by pyknosis and eosinophilic granulation of the cytoplasm in the renal cortex was also observed. One mouse had R-E cells and lymphoid hyperplasia and a number of megakaryocytes in the spleen. b. Gross changes in later deaths and unscheduled terminations: Mice that died or were terminated before schedule were generally in poor nutritional condition with little or no fat deposits. A number of lesions were noted in these mice as well as those terminated as scheduled. These lesions included small, raised tan to grey nodules varying from pinhead size to 0.5 cm in diameter, and small dark hemorrhagic spots from petechiae to 1 cm in diameter in the lung. Small, dark nodular masses In the liver as well as internal hemorrhage into the abdominal cavity were observed in some of the mice. Most of the livers were moderately to severely mottled in appearance. The mammary tumors found were of varying size and were greyish white to dark red in color. 17 SL 083832 c. Turn rs In mice exposed to VC: Microscopic examination of the mice that died or were terminated, both scheduled and unscheduled, revealed a variety of tumors. The tumor occurrence during 1 to 3 months, 4 to 6 months, 7 to 9 months, and 10 to 12 months are summarized in Tables 67 to 72. The cumulative incidence of all tumors is summarized in Table 73. Bronchiolo-alveolar adenomas in mice first occurred during the 2nd month of exposure (Table 67). At this time, one male and two fe males exposed to 1,000 ppm and one male exposed to 250 ppm were found to have bronchiolo-alveolar adenomas. After 3 months exposure, one male at 50 ppm, one male at 250 ppm, and two males and one female at 1,000 ppm were found to have bronchiolo-alveolar adenomas. Between 4 and 6 months, two males at 50 ppm, one male and four females at 250 ppm, and six males and four females at 1,000 ppm had bronchiolo-alveolar adenomas (Table 68). Be tween 7 and 9 months, two males and four females at 50 ppm, five males and eight females at 250 ppm, and 13 males and 19 females at 1,000 ppm developed this tumor (Tables 69-71). During the final 3 months of exposure, three males at 50 ppm and two males at 250 ppm were found to have bronchiolo alveolar adenomas (Table 72). The bronchiolo-alveolar adenoma was charac terized by papillary proliferation of alveolar epithelium forming small and well-demarcated, but not encapsulated, round nodules. The severity of this tumor was in direct proportion to the level of VC and length of expo sure. In the more severe lesions, there was an increase in number and size of the nodules by expansion and coalescing to cause consolidation of the affected lobes of some animals. These bronchiolo-alveolar adenomas occurred in a total of eight males and four females exposed to 50 ppm, 10 males and 12 females exposed to 250 ppm, and 22 males and 26 females exposed to 1,000 ppm (Table 73). Only one control male mouse was found to have a bronchiolo alveolar adenoma during the 9th month. Hemangiosarcoma of the liver was first seen in mice during exposure for 4 to 6 months. Two females exposed to 250 ppm and two males and three females exposed to 1,000 ppm had hemangiosarcomas of the liver during this period (Table 68). Between 7 and 9 months, one male at 50 ppm, five males and 14 females at 250 ppm, and 11 males and 15 females at 1,000 ppm were found to have hepatic hemangiosarcomas (Tables 69-71). Be tween 10 and 12 months, additional hemangiosarcomas of the liver were found in two males at 50 ppm and two males at 250 ppm (Table 72). These hemangio sarcomas were characterized by mild to severe proliferation of endothelial cells lining the sinusoids, dilation of the sinusoids, focal hemorrhage forming small to large cavernous blood spaces, Invasion of the hepatic parenchyma with neoplastic tissue and mild to severe necrosis. Hepatic hemangiosarcomas occurred in a total of three males exposed to 50 ppm, seven males and 16 females exposed to 250 ppm, and 13 males and 18 females exposed to 1,000 ppm (Table 73). This tumor was not found in any control mice. A few hemangiosarcomas were also found in tissues other than liver. These 18 SL 083833 tissues included the mammary gland, heart, skeletal muscle, gastrointestinal tract, kidney, pancreas, mesenteric lymph nodes, epididymus, testes, and mesentery (Table 73). The severity of hemangiosarcoma in the liver and the overall incidence in other tissues were related to the level of VC and the length of exposure. The third major type of tumors was found in the mammary gland of the female mice exposed to VC. These tumors were composed of ductular adenocarcinoma, squamous cell carcinoma, and/or anaplastic carcinoma. Between 4 and 6 months, one and three females exposed to 250 or 1,000 ppm, respectively, had mammary gland tumors (Table 68). Between 7 and 9 months, seven, two and 10 females exposed to 50, 250 or 1,000 ppm, respectively, developed these tumors (Tables 69, 70 and 71). Between 10 and 12 months, mammary gland tumors occurred in two females exposed to 50 ppm (Table 72). No females exposed to 250 or 1,000 ppm survived during this period. The adenocarcinoma was charac terized by proliferation of ductular epithelium with marked anaplastic and squamous cell metaplasia; the squamous cell carcinoma was characterized by marked proliferation of stratified squamous epithelium, marked keratlnization, marked purulent inflammation and necrosis; and anaplastic carcinoma was char acterized by marked proliferation of undifferentiated cells in large sheets, irregular cords and packets. These mammary gland tumors occurred in nine, three and 13 female mice exposed to 50, 250 or 1,000 ppm, respectively. Most of these mice had more than one of these different tumors in the mammary gland. These tumors were more severe in the mice exposed to higher levels of VC and in the mice that died or were terminated at a later date. In addition, the squamous and/or anaplastic cell carcinomas metastasized to the lung of a number of mice. This type of tumor was not seen in control female mice (Table 73). Malignant lymphoma was seen in one female mouse exposed to 50 ppm during the 6th month and was characterized by marked disseminated or diffused infiltration of lympho-reticular cells in the epicardium and myo cardium, perivascular and interstitial areas of the lung, liver, spleen and kidney. There was loss of splenic architecture. In addition, one male ex posed to 1,000 ppm had a malignant lymphoma characterized by a large mass of lympho-reticular cells and necrotic debris, Infiltrating the cervical tissue surrounding the trachea, blood vessels and esophagus. During the 9th month, two females exposed to 250 ppm and one male and three females exposed to 1,000 ppm had malignant lymphomas involving spleen, liver, lung, heart, subcutaneous tissue in the cervical area, and/or mammary gland. Malignant lymphoma was not found in any control mice or mice exposed to 55 ppm of VDC at any time. There was hemangioma, renal adenoma or skin keratocanthoma in one mouse each exposed to VC (Table 73). 19 SL 083834  d. Tumors In mice exposed to VDC: In mice exposed to 55 ppm VDC, a few small nodules of bronchiolo-alveolar adenoma occurred in one male during the 6th month, two males during the 9th month, and three males during the 12th month, Hemangiosarcoma occurred in the liver of one male during the 9th month, and one male and one female during the 12th month. One male had a hepatoma when terminated during the 9th month, one female developed a hepatoma during the 12th month. The hepatomas were characterized by a marked proliferation of hepatocytes with a loss of the lobular pattern in the liver, except for the male terminated during the 12th month. This mouse had only a tiny focus of the neoplastic cells. Hepatoma was not found in any control mice or mice exposed to any level of VC. Hepatic cell carcinoma or renal adenoma occurred in one male each exposed to VDC during the 12th month. Mammary gland tumors were not found in any mice exposed to VDC. The cumulative incidence of tumors in mice exposed to VDC is sum marized in Table 73. e. Other lesions: As seen in rats, there were a number of spontaneous or incidental lesions in various tissues of the control mice and mice exposed to VC or VDC for various lengths of time (Tables 74-79). These lesions were not related to VC or VDC. However, there were a few lesions in the liver of mice that appeared to be related to VDC. The livers of mice exposed to 55 ppm VDC for 9 or 12 months had microfoci of mononuclear cells, focal degeneration and necrosis, intranuclear inclusions and/or large basophilic nuclei (Tables 78 and 79). While these lesions also appeared in some controls, the incidence and/or severity was greater in the mice exposed to VDC. In addition, there was an increased incidence of amyloidosis in the kidney of mice exposed to VDC (Table 79). The myeloid/erythroid (M/E) ratios of the bone marrow smears of control mice and mice exposed to VC or VDC were within normal limits at all time periods examined (Tables 74-79). C. Special Studies 1. ^C-Thymldine incorporation into DNA in mice: Histopathologic examination had revealed a moderate number of mitotic figures in the livers of some mice exposed to VC during the 9th month. The DMA synthesis was undertaken to determine if this observation could be confirmed at the bio chemical level. DNA synthesis, as measured by ^C-thymidine incorporation into DNA, was significantly increased in the livers of male mice exposed to 50 ppm VC for 11 months (Table 80). However, the DNA concentration, expressed as mg DNA/g of liver, was not significantly affected. Histopathologic examination of these livers used in this study did not reveal an increase in the number of mitotic figures or any evidence of neo- or preneoplastic lesions. 20 SL 083835 2. H11eme s.y..n..t.h...e..s..i.s.. in rats: A number of chemicals are known to Oo / produce porphyria in both man and animals by disrupting porphyrin metabolism.--' To determine whether VC affects porphyrin metabolism some aspects of the heme biosynthetic pathway were studied in rats exposed to VC for 12 months. Hepatic levels of ALAS were determined since it is the rate limiting step in heme biosynthesis and the activity of ALAS is increased in drug-induced porphyria. Urinary levels of ALA were also determined because epoxides of VC may combine with essential sulhydryl groups on ALA dehydrogenase in a manner similar to lead. Inactivating this en2yme may result in an increased excretion of ALA. In addition, urinary ALA excretion, which is used to de tect lead poisoning, might also be used as a monitoring system for detection of VC intoxication if heme synthesis was affected by VC. The studies of ALAS activity in liver of rats exposed to 50 or 250 ppm VC for 12 months showed no significant effect of VC (Table 81). The ALAS activity observed in this study was about 50% of the activity previously reported by Woods,--^ Although there was some contamination of urine samples with drinking water, the general pattern of results suggest that VC exposure does not alter the urinary excretion of ALA. The results of this study were expressed on an arbitrary scale since the low level of ALA prevented an accurate determination of ALA concentration (Table 82). After the addi tion of Ehrlich's Reagent, all samples produced a faint yellow to pink color while none of the samples had a distinct red color. 3. Dominant lethal studies in rats: The purpose of this study was to determine if repeated exposures to VC or VDC, for nearly 3 months, produced germinal mutations, of the dominant lethal type, in male rats. VC exposure did not significantly reduce the ratio of fertile to tested males. However, there was a reduced number of males exposed to 1,000 ppm VC that mated with two females (Table 83). In addition, there was a re duced ratio of pregnant to mated females in the groups mated with males exposed to 250 or 1,000 ppm VC or 55 ppm VDC. The mating period ranged from 7 days for the control group to 8 to 9 days for the treated groups. Pregnancies that resulted from the mating of exposed males with unexposed females were normal in terms of corpora lutea and implants (Table 84). In these females, there was no evidence of preimplantation loss, as measured by the ratio of implants to corpora lutea, or postimplactation loss, as measured by the ratio of viable implants to total implants. 4. Alpha-fetoprotein: The appearance of an embryo-specific globulin (a-fetoprotein) in serum has been reported in adult animals and humans with primary hepatomas.--^ In order to determine whether a-fetoprotein could serve as an early indicator of hepatic hemangiosarcoma, serum or plasma 21 SL 083836 from rats and mice were assayed for ot-fetoprotein levels. The serum or plasjna was obtained from animals terminated at various periods as well as from a number of unscheduled terminations. Levels of a-fatoprotein from rats ex posed to VC for 9 months showed no differences from controls (Table 85). In mice exposed to VC or VDC, occasional high levels of a-fetoprotein occurred (Table 86). Two (Nos. 177 and 240) of the mice with elevated afetoprotein levels had hepatic hemangiosarcomas and the third mouse (No. 324) had a hepatoma. However, the a-fetoprotein level of several other mice that had hepatic hemangiosarcoma was not elevated. 22 SL 083837 rv. Discussion and conclusions A. Rats 1. Vinyl chloride: Exposure of rats of both sexes to 50, 250 or 1,000 ppm VC was without adverse effects through 7 months of exposure. Beginning with the 8th month, a number of rats died or were terminated before schedule. These deaths and early terminations occurred primarily in the 250 and 1,000 ppm VC exposed groups. The body weights of the male and female rats exposed to VC were comparable to controls until after 43 weeks of exposure. At this time, the weight gains of both males and females exposed to 1,000 ppm VC were less than controls. The exposure to VC did not result In any persistent changes in hematology, clinical blood chemistry, cytogenetic parameters, collagen content in liver or lung, pulmonary macrophage count or the bones of the extremities. Histopathologic examination of rats exposed to VC for up to 12 months showed a number of tumors. The tumors found with the greatest fre quency were hemanglosarcomas of the liver and lung. Hemangiosarcoaa of the liver started to appear during the 9th month. It occurred In two males and 10 females exposed to 250 ppm VC and in six males and 15 females exposed to 1,000 ppm. Most rats with hepatic hemangiosarcoma also developed hemangiosarcoma in the lung. Hemangiosarcoma of the lung was found in three females exposed to 250 ppm and four males and nine females exposed to 1,000 ppm. The occurrence of these tumors is dose and time related; the occurrence in females was greater than in males. Hemanglosarcomas were also found in other tissues including skin, omentum, mesentery, and mesenteric lymph nodes. Malignant lymphoma was found in one female rat exposed to 250 ppm VC and one male and two females exposed to 1,000 ppm VC. The occur rence of the malignant lymphomas may be related to the VC exposure. A number of spontaneous or incidental tumors were found in various tissues. These tumors occurred in one or a few rats and were not related to the exposure to VC.2 2. Vinylldene chloride: Exposure of rats to 55 ppm VDC for up to 12 months had very few adverse effects. The weight gain of both males and females were less than controls during the final 3 months of exposure. No consistent changes in hematology, clinical blood chemistry, cytogenetic analysis, collagen content of liver or lung, pulmonary macrophage counts or in the bones of the extremities were found. 23 SL 083838 One male rat exposed to 55 ppm VDC was found to have a hemangiosarcoma of the mesenteric lymph nodes and another male had a hemangiosarcoma of the skin. These tumors may be related to the VDC exposure. A few lesions occurred in the livers of rats exposed to 55 ppm VDC. Fatty changes of the liver appeared during the 9th month; nodular or disseminated vacuolization occurred during the 12th month. These lesions were more severe and the incidence was greater in rats exposed to VDC. It appears that these lesions were related to VDC. B. Mice 1. Vinyl chloride: a. Acute effect and general observation: Exposure to VC caused deaths of two males and one female exposed to 1,000 ppm VC between the 3rd and 9th days. Histopathology revealed acute toxic hepatitis and marked tubular necrosis of the renal cotex. Thereafter, the mice appeared to be in good health. During the 6th month, the general health of a few mice began to deteriorate. They died or were terminated. During the 7th month, the health of these mice worsened. By the end of the 9th month, all males and females exposed to 1,000 ppm VC and all females exposed to 250 ppm VC had died or were terminated before scheduled. At the conclusion of 12 months only the controls and a few mice exposed to 50 ppm VC had survived. The clinical signs included rough hair coat, lethargy, sudden weight loss, and the development of external tumor masses and/or abdominal distensions. Nearly all of the mice that died or were terminated ahead of schedule and many of the mice terminated at the scheduled times had one or more types of tumors. b. Bronchiolo-alveolar adenoma: Bronchiolo-alveolar adenoma was first noted in mice exposed to 250 or 1,000 ppm VC for 2 months and in mice exposed to 50 ppm VC after 3 months exposure. In the period between 3 and 12 months of exposure to VC, bronchiolo-alveolar adenomas were found in 37% of the mice exposed to 50 ppm VC, in 51% of the mice exposed to 250 ppm VC, in 89% of the mice exposed to 1,000 ppm VC. The overall in cidence and severity of the tumor was directly related to the level and length of exposure. Only one bronchiolo-alveolar adenoma was found in the control mice and this occurred in one male during the 9th month of exposure. Bronchiolo-alveolar adenoma, bronchiolar adenoma, or pulmon ary adenomatosis, in mice have been reported to occur spontaneously in aging mice, mostly over 1 year of age.2.fr~/ However, in our study, large numbers of mice exposed to various levels of VC developed this tumor. In addition, the tumor started to occur at a very early age and had a dose and time re lationship as regard to the incidence and severity. On the other hand, 24 083S39 only a few small nodules of this tumor occurred in several mice treated with VDC. In addition, they occurred relatively at a later time. Its significance In these mice is questionable. c. Hemangiosarcoma: Hemangiosarcoma of the liver first occurred in mice exposed to 250 or 1,000 ppm VC for 5 to 6 months. After the 6th month, approximately 75% of the mice examined in the 250 and 1,000 ppm VC exposed groups had hepatic hemangiosarcomas. The occurrence of the hepatic hemangiosarcoma appeared to be greater in the female mice than in the males. In the mice exposed to 50 ppm VC, three male mice were found to have hepatic hemangiosarcomas. In addition, hemangiosarcomas were occasionally found in other tissue including mammary gland, heart, skeletal muscle, gastrointestinal tract, kidney, pancreas, mesenteric lymph nodes, epididymis, testes and mesentery. The incidence and severity of the hemangiosarcoma in the liver and the overall incidence of hemangiosarcoma in other tissues were related to the exposure level of VC and the length of exposure. No hemangiosarcomas were found in any tissues of control mice. d. Mammary tumors: Mammary tumors were found in females exposed to all levels of VC. The tumors consisted of ductular adenocarcinoma, squamous and anaplastic cell carcinomas. The mammary gland tumors were first found after 6 to 7 months. The incidence and severity of the tumors appeared to be greater in mice exposed to higher levels of VC and in those mice ex posed for the longest periods of time. This may explain the higher inci dence of mammary tumors in the group exposed to 50 ppm as compared to the group exposed to 250 ppm. All females exposed to 250 ppm had died by the end of 9 months, while many females exposed to 50 ppm survived beyond this time. This increased exposure time may thus account for the greater number of mammary tumors. As mentioned above, a number of hemangiosarcomas were found in the mammary gland. In addition, the squamous and/or anaplastic cell carcinomas of the mammary gland also metastasized to the lung. No mammary gland tumors were found in control mice or in male mice. The pathogenesis of the mammary tumor was of a complex type. Observations of various stages and sizes of the mammary tumors suggest that the tumor originated as ductular adenocarcinoma and then in a very early stage underwent an anaplastic and squamous cell metaplasia. At this point, the tumor was quite malignant and invasive. In many cases, there was metastasis of squamous and/or anaplastic cell carcinoma to the lung. In addition, the ductular or alveolar involvement or pattern in the mammary gland, characteristic of spontaneously occurring mammary tumors in mice, was minimal except in the early stages of some small tumors. This pro vides further evidence that the mammary tumors are related to the VC ex posure. 25 SL 083840 e. Malignant lymphoma; Malignant lymphoma was found in one female exposed to 50 ppm VC, two females exposed to 250 ppm and in two males and three males exposed to 1,000 ppm VC. Generally, the lymphomas appeared to be disseminated, affecting a number of var . us tis sues. No malignant lymphomas were found in control mice. f. Other tumors: There were other tumors occasionally found in mice but these tumors were considered to be spontaneous and un related to the VC exposure. Other observations: No consistent changes were found in mice exposed to VC when compared to controls in hematology, clinical blood chemistry, cytogenetic analysis of bone marrow cultures, or X-ray examination of bones from the extremities. The pulmonary macrophages count was elevated in mice exposed to 250 or 1,000 ppm. The mice with elevated macrophage counts also had bronchiolo-alveolar adenoma. However, not all mice with bronchiolo alveolar adenomas had the elevation of macrophage count. The significance of this observation is not clear. 2. Vlnvlldene chloride; Mice exposed to 55 ppm VDC for up to 12 months were found to have some tumors including bronchiolo-alveolar adenoma, hepatic hemangiosarcoma and hepatomas. The bronchiolo-alveolar adenomas were found in six male mice. Bronchiolo-alveolar adenoma, bronchlolar adenoma or pulmonary adenomatosis has been reoorted to occur spontaneously in aging mice, mostly over 1 year of ageSince most of the bronchiolo-alveolar adenomas in VDC exposed mice occurred in the late stages of exposure, the relationship of these tumors and the VDC ex posure is questionable. Hemangiosarcoma of the liver was found in two males and one female exposed to 55 ppm VDC, and is probably related to the VDC exposure. In ad dition, one male and one female mouse exposed to VDC developed hepatomas. The significance of this tumor and its relationship to VDC exposure is ques tionable. Hepatomas have been reported to occur spontaneously In small num bers in mice approximately 1 year of age.2.Zri9/ Therefore, it is possible that the hepatomas found in this study are of a spontaneous nature since they occurred latex in the study. There were no hepatomas found in control mice. A number of other lesions also occurred in the liver of mice ex posed to 55 ppm VDC. These lesions Included enlarged and basophilic hepa- tocytes, enlarged nuclei with eosinophilic inclusions, mitotic figures or polyploidy, microfoci of mononuclear cells, focal degeneration and necrosis. It may be that these lesions are ,,preneoplastic', and may be related to the occurrence of hemangiosarcoma or other liver tumors. Since only a few mice developed liver tumors compared to the large number with the "preneoplastic" lesions, further study of VDC in mice utilizing additional exposure levels is needed to establish any definite relationship between the VDC exposure, the "preneoplastic" lesions, and subsequent development of neoplastic lesions in the liver. ^ SL 083841 C. Special Studies 1. ~^C-Thymidine incorporation into DNA in mice: An increased incorporation of ^C-thymidine into liver DNA was found in male mice exposed to 50 ppm VC for approximately 11 months. Disregarding pool size considera tions, this suggests that there was an increased population of dividing cells in the livers of these mice. An increased population of dividing cells could be due to either regenerative or carcinogenic processes. Due to the low inci dence of hepatic tumors at 50 ppm VC, it would seem most likely that the in creased thymidine incorporation into DNA is the result of regenerative pro cesses. However, further study would be needed to confirm this observation. As previously mentioned, the study of the thymidine incorporation into DNA was undertaken in response to the observation of the increased numbers of mitotic figures in the livers of mice exposed to VC. While in creased incorporation of ^C-thymidine into DNA was observed in the present study, histopathologic examination did not reveal an increased number of mitotic figures in the liver of these mice. This would indicate that bio chemical changes may provide a more sensitive monitor for VC related effects than do morphological changes. 2. Heme synthesis in rats: Studies of hepatic ALAS activity in the liver and urinary levels of ALA did not show any effects related to the VC exposure. These results would indicate that VC, under the conditions of this study, does not have an effect on the heme biosynthetic pathway. However, the possibility that long-term exposure to VC may result in an adaptation to an acute effect of VC on the heme pathway should be considered. 3. Dominant lethal studies in rats: Neither preimplantation nor postimplantation losses were observed in pregnancies that resulted from the mating of VC or VDC exposed male rats. However, the occurrence of a preimplantation loss was suggested by the increased number of nonpregnant fe males in the groups mated with males exposed to 250 or 1,000 ppm VC and 55 ppm VDC. The relevance of this observation to the detection of a dominant lethal effect is of questionable significance for two reasons. First, there was no indication of a preimplantation loss in pregnant rats. Secondly, a postimplantation loss is more indicative of dominant lethality than a preimplantation loss. Since neither prelmplantation nor postimplantation losses were observed in pregnant rats, it is concluded that the VC and VDC exposures utilized in this study did not produce dominant lethal mutations in the germinal cells of male rats.4 4. Alpha-fetoprotein levels in rats and mice: The assay of afetoprotein levels in the serum or plasma of rats and mice does not appear to serve as a useful indicator of VC or VDC induced hepatic hemangiosarcoma. Only three mice were found to have elevated a-fetoprotein levels. Two of these mice had hepatic hemangiosarcomas and the third mouse had a hepatoma. However, an additional 10 mice with hepatic hemangiosarcoma did not have elevated a-fatoprotein levels. 27 SL 083842 ACKNOWLEDGEMENTS We gratefully acknowledge the technical assistance of Dr. Stewart Sell, Department of Pathology, School of Medicine, University of California at San Diego, LaJolla, California, for the a-fetoprotein assays, and Dr. Norman L. Martin, Department of Diagnostic Radiology, University of Kansas Medical Center, Kansas City, Kansas for the radiographic examination of the rat and mouse limbs. 23 SL 083843 REFERENCES 1. Lee, C. C. , et al.: Toxicological Evaluation of Ferric Dimethyldithiocarbamate (Ferbam) and Dithiocarbamate (Thiram) with Acute Toxicity of Manganese and Zinc Ethylenebisdithiocarbamates (Maneb and Zineb). NIH Contract No. N01-ES-2-2Q84 (Continuation of NIH-NIEHS-72-2084), 1-356, 1975. 2. Patty, F. A., W. P. Yant, and C. W. Waite: Acute Response of Guinea Pigs to Vapors of Some New Commercial Organic Compounds V: Vinyl Chloride. Pub. Health Rept., 45: 1963-1971, 1930. 3. Oster, R. H., C. J. Carr, J. C. Krantz, and M. J. Sauerwald: Anesthe sia XXVII, Narcosis with Vinyl Chloride. Anesthesiology, 8: 359-361, 1947. 4. Carr, J., R. M. Burgison, J. F. Vitcha, and J. C. Krantz, Jr.: Anesthe sia XXIV, Chemical Constitution of Hydrocarbons and Cardiac Automaticity. J. Pharmacol., 97: 1-3, 1949. 5. Mastromatteo, E., A. M. Fisher, H. Christ!, and D. Danziger: Acute Inhalation Toxicity of Vinyl Chloride to Laboratory Animals. Am. Ind. Hyg. Assoc. J., 21: 391-397, 1960. 6. Torkerson, T. R., F. Oyeri, and V. K. Rowe: The Toxicity of Vinyl Chloride as Determined by Repeated Exposure of Laboratory Animals. Amer. Ind. Hyg. Assn. J., 22: 354-361, 1961. 7. Viola, ?. L., A. Bigotti, and A. Caputo: Oncogenic Response of Rat Skin, Lungs, and Bones to Vinyl Chloride. Cancer Research, 31: 516-522, 1971. 8. Maltoni, C. and G. Lefemine: The Potential of Experimental Stages of the Prediction of the Ambient Oncogenic Risks. An Example: Vinyl Chloride. Lincel-Rend. Sc. fs. mat. enat., 56: 1, 1974. 9. Quick, A. J.: A Study of the Coagulation Defect in Hemophilia and in Jaundice. Am. J. Med. Sci., 190: 501, 1935. 10. Jendrassik, L., and P. Grof: Simplified Photometric Method for the Determination of the Blood Bilirubin. Siochem. Z., 297: 81, 1938.11 * * * 11. Mancini, G., A. 0. Carbonara, and J. F. Heremans: Immunochemical Quantitation of Antigens by Single Radial Immunodiffusion. Immunochemistry, 2: 235-254, 1964. V SL 083844 29 12. Kingsley, G. R.: The Direct Biuret Method for the Determination of Serum Protein as Applied to Photoelectric and Visual Colorimetry. J. Lab. Clin. Med., 27: 840, 1942. 13. Doumas, B. T., W. A. Watson, and H. G. Biggs: Albumin Standards and the Measurement of Serum Albumin with Bromcresol Green. Clin. Chim. Acta, 3'-W1: 87, 1971. 14. Tjio, J. H. and J. Wang: Direct Chromosome Preparations of Bone Marrow Cells. In: Human Chromosome Methodology, J. J. Tunis (ed.), Academic Press, New York, 1965. 15. Moorhead, P. S. and P. C. Norwell: Chromosome Cytology. In: Methods in Medical Research, H. N. Eisen (ed.), Year Book Medical Publ. Inc., Chicago, 10: 310, 1964. 16. Dunnett, C. W.: A Multiple Comparison Procedure for Comparing Several Treatments with a Control. J. Am. Stat. Assoc., 50: 1096-1121, 1955. 17. Burton, K.: A Study of the Conditions and Mechanisms of the Diphenylamine Reaction for the Colorimetric Estimation of Deoxyribonucleic Acid. Biochem. J., 62: 315, 1956. 18. Woods, J. S.: Studies on the Role of Heme in the Regulation of a-Amino- levulinic Acid Synthetase During Fetal Development. Molec. Pharmacol., 10: 387-389, 1974. .-V 19. Lowry, 0. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall: Protein Measurements with Folin Phenol Reagent. J. Biol. Chem., 193: 265-275, 1951. 'v~ 20. Davis, J. R. and S. L. Andelman: Urinary a-Aminolevulinic Acid (ALA) Levels in Lead Poisoning. II: A Modified Method for the Rapid Deter mination of Urinary a-Aminolevulinic Acid Using Disposable Ion-exchange Chromatography Columns. Arch. Environ. Health, 15: 53-63, 1967. 21. Sell, S. and D. Gord: Rat a-Fetoprotein III. Refinements of Radio immunoassay for Detection of 1 ng Rat <*]_F Imaunochemistry, 10: 439442, 1973.22 * 22. Tschudy, D. P. and H. L. Bonkowski: Experimental Porphyria. Fed. Proc., 31: 147-159, 1972. 30 SL 083845 23. Smith, J. B.: a-Fetoprotein: Occurrence in Certain Malignant Diseases and Review of Chronical Applications, Med. Clin. N. Amer. 54: 797803, 1970. 24. Baillif, R. N. and J. L. Jones: Pulmonary Adenomatosis in Aging Mice. J. Comp. Path. 83: 597-603, 1973. 25. Amaral-Mendes, J. J.: Histopathology of Primary Lung Tumors in the Mouse, J. of Pathol. 97: 415-422, 1969. 26. Deerberg, F. et al.: Der Lungentumor der Maus: ein progressive Tumor, Vet. Path. 11: 430-441, 1974. 27. Sanford P. Sher: Tumors in Control Mice: Literature Tabulation, Toxicol. Appl. Pharmacol., 30: 337-359, 1974. 28. Percy, D. H. and A. N. Jonas: Incidence of Spontaneous Tumors in CD-1 HaM/ICR Mice, J. Nat. Cancer Inst., 46: 1045-1065, 1971. 29. Andervont, H. B.: Studies on the Occurrence of Spontaneous Hepatomas in Strains C3H and CBA, J. Nat. Cancer Inst., 11:. 581-592, 1950. 30. Mann, H. B. and D. R. Whitney: On a Test of Whether One or Two Random Variable is Statistically Larger than the Other, Ann. Math. Scat., 18: 50-60, 1947.31 31. Siegel, S.: Nonparametric Statistics, McGraw-Hill, New York, 96-104, 1956. SL 083846 31 TABLE 1 LABORATORY DATA OF MALE RATS AFTER EXPOSURE TO VC OR VPC FOR 1 MONTH CRYfHffOCTTCS (iu /mm ) flCTICULOCrrcs, ft nn*rOC*tT, vOL* 1 hC^OGLOAIN* Oh, ft mcv. cubic hichons MCHB* MICRO MICROQNS* HCMBC* Of* 1 53 PLATELETS (Xio /mm ) 33 LEUKOCYTES (XU /mm ) HCUTAOPHtLS* 1 LTHRwOCyTCS* ft SAMOS* ft eosinophils* ftASOAHlLS* ft hohocttcs* ft ATYPICAL. ft NUCLEATED ABC* ft HROTNROHBlN TIMCt SEC* SOOT, IU/L SOPT* WL al*. pnos.* UJ/L aiLiRoerw* total ** * ylLIPUfllN, DIRECT MG ft SUN* G ft CREATININE* M ft LOH IU/L THHUMOOLOBUCINS A* IU/L 3* a factor* iu/nl 3. FACTOR* IU/NL N* JU/Ht total protein* h ft ALftOHtN* OH ft OLOHULlNt OH ft 3 HACftOPHAOeS (NO./HH ) 6,40 .5* (3) *?! 1 .25 (3) *4.0 & .4 (3) 15.1 r *2 (3) AS.5 1 *.4 <3) 22** 1,6 C3> .3**2 T.* ,0 (3) .7 (3) n*7 - ,4 (3) 14.3 1 1.3 *3.5 V 2.4 0*0 L 0.0 .s 1 .3 0*0 1 0.0 v* 1 .0 0*0 1 0,0 0.0 1 0.0 12.2 & 3 03.3 & 9.3 30.3 W* 03 3 9 l i ,1 0.0 0.0 14.0 1*1 .7 *1 513 - 307 7.09 * 1 .34 * *4,3 , .13 52 .4 15.0 * .2 42.* * 21.2 . 33*9 . 6.2 11.4 * 1*.3 1.0 .2 .2 .0 1.5 3.2 03*0 L *.4 0.0 a 0.0 1 .4 1.2 0.0 0.0 1.0 * .7 0*0 . 0.0 0*0 * 0.0 11.4 * *4 45.0 * 3*5 32*5 * 1.5 l 124 * 0 * *3 .1 .1 .0 17,$ # ,9 .* * 0.0 239 * 104 4*2 1 343 .4 42 7ft * *7 22 2 9.3 * .3 3*3 * *2 1 .9 .2 u* 43 5*4 . 3*5 1.4 2 *L .2 VC HO pp* 7.*4 l *4 *5.0 t 15.2 * 0* .*7 .* .1 40*3 1 .7 20.* i .3 33*4 .2 0.2 .4 9,0 9,5 i .5 1.4 99.4 & 1.4 0*0 1 0*0 3 i .3 0*0 1 0*0 .3 i *3 0*0 0.0 0*0 0*0 11.9 9 .1 44.5 1 2*9 33.3 l* 9ft 9 .3 1 .2 i 13.4 9 .1 .t i' .9 .4 0.0 397 1*9 5*4 * 3*7 2.0 .1 .2 .1 7C 1,000 6*04 2.30 * *4 1,1* *2*5 * 1*.* # .9 .3 71.7 4.2 24*2 * 33.9 1.9 * 4*7 * 1.1 11.3 . 1.1 14.3 # 5.* 40.3 0.0 * 3.7 0.0 1.0 * *4 0.0 0*0 *ft * 0*0 . .3 0*0 0.0 . 0*4 12*1 * *3 44*5 * 1*9 3**0 * 102 . 1*2 ft *3 - *1 .1 .0 15.3 ,4 4 0.0 302 . 10* 5*5 340 * .1 20 107 30 * 24 1 3.4 3.3 * 2.* . 1 *3 *3 *39 93 EuctIm r m*aa i S.t. f 4 ftc setpc * aoc*4 is patmchM**. / t*vm eh* Coocrol $tv*9, 9 < 0.05, amulet'* oulclpl* ca*prtien proctdur*. 5v5 j>p 7.31 1.17 *< 3 1*.3 * #06 10 >9 *2 40*3 # 20,* * .3 33.8 * . 5.2 ,0 v, a l*o - .6 2.7 43*3 0.0 2*o 0.0 1*0 * 0.0 0.0 . 0.0 t.a u.o *3 0.0 0.0 L 12.1 . 95.9 * 0.0 A 34.5 * U4 103 * 6 .3 . .L .2 .1 14.9 . 1.1 *7 . *1 309 . 199 5*9 *1 3,9 V.} 1 .1 243 i 154 32 SL 083847 TABLE 2 LABORATORY DATA OF FEMALE RATS AFTER EXPOSURE TO VC OR VDC FOR l MO>rTH 63 ERTTHflOCTTCS {*10 /MN > rftTICULOCVTESt 1 fMA.TOCJlT VOL, ft hCNOOLOBIN. IjM, mcv* cubic microns MCKB. MICRO MtcROGNS. MCHBC* GM ft 53 PLATELETS 1X10 /mm \ 23 LEUKOCYTES (X10 /mm \ NEUTROPHILS* wTMp*OCYTS* ft 34*405* ft EOSINOPHILS* ft 94S0PHILS# ft MONOCYTES* ft atypical* ft nucleated wee. ft PflOrwflONQlM TtwC* SEC# SOOT* IU/L SGPT, IU/L ALK, PMQS** rU/L 91LIRUBIN* TOTAL mo ft BILIRUBIN* DIRECT MO ft 3UN. MO ft CREATININE* mg ft L0H* iu/L Csacral 5.19 * .3* 1*57 .67 *1.5 .5 l*.5 i .2 57,9 * * * 7 23.7 * 1.7 3**9 * .1 6.5 i .5 7.3 i .5 17,3 3.1 91.9 3.1 0.0 0.0 .4 1 .5 0.0 1 0.0 3 .3 0.0 0.0 0.0 0*0 12*1 ? -3 < 7*.3 9,9 33,5 * 2.9 74 * 7 1 .1 *1 .1 15*5 1.6 6 0.0 391 215 VC 30 ?tm 6.55 *21 1.34 .53 *3*0 * 0.0 1 * 7 *2 64.9 2.0 22*2 .9 34.2 *4 5.5 . 4 7.3 .9 10*0 2*9 99.5 3*3 0.0 0.0 1.0 .4 0.0 0*0 *5 1 *3 0*0 0.0 0*0 0*0 11.5 *2 55.4 10*7 27.0 2*1 55 *2 1 1 1 .1 15*0 1*2 *6 0.0 404 234 7C 230 ?l 5.22 *19 1*17 *35 *i.a 14.2 .6 .3 67*2 * 1.3 22*9 *2 34,1 .3 7.5 .9 7.7 * .5 13.5 4,6 54*3 *1 0.0 0*0 .9 ,5 0.0 0.0 *5 *3 0.0 0.0 0.0 0.0 11.6 .2 71*3 6*9 27.4 1.4 77 LQ *2 .1 *1 16*0 .1 .7 *7 .1 *64 147 IMMUNOGLOBULINS A* IU/ML G* A FACTOR. IU/**L o* e factor* iu/nl m* Iu/*L TOTAL PROTEIN. 9M ft ALBUMIN* 9 ft GLOBULIN* ON ft 3 maCRORnaOCS (NO./mm i 7,3 .4 343 * 3* 71 12 44 * 3 5.3 *1 3*4 2.1 *1 *1 24 72 5*7 3.7 2*0 * l *2 *1 5*9 3.9 2*0 .2 *2 .1 M>C4 LaEaeflo art mas : J.I. of 4 rati me ye i< SignlficAaclr dlffarvne fro* eft* C*sc?ol froup, p < 0,05, DtmsAce'* ouleiplc eofortooo procaduv*. A, 1,000 ?P* 5.47 * ,51 1.76 * .77 *0.0 * . * l*.l .1 70,0 7,4 2*.9 2.4 35*2 3 7.9 * ,a 5*9 .5 15.3 3*3 43.0 3.4 0*0 0.0 .3 3 0.0 * 0.0 *5 *3 0.0 0.0 0.0 0.0 11*7 9 61*0 2.* 29.3 * 3.1 *0 2 *2 .1 .0 *0 15.3 * .9 .6 0.0 72 6 7*1 9 399 4 1 SO 13 *9 i 5.7 * 2 .1 3*5 .2 2.2 .2 293 93 'DC 55 ;p" 6.73 * 19 94 * l i *0,0 14,3 59.5 20.4 3*.9 i .2 .6 *2 .2 5.5 1 U .9 12.a - .6 ,1.0. a" 3.0 96.0 - 2. 7 0.0 .9 - 0.0 .5 - 0,0 .1 0*0 ,1 0.0 - 0.0 0.0 - 0.3 11.2 - -l (2) 73,3 - 9*3 (3) 3*. 0 - 1.7 (1) 60 5 O) .3 - * l '3> ,2 - - - 31 1 7,3 - 1-P >2) 1.0 i .1 ;3l* 132b 166 1') r .i O)1 3.9 * ,,* ; .3 (3) .3 (31 269 a *5 SL 083848 33 TABLE 3 LABORATORY DATA OF MALE RATS AFTER EXPOSURE TO VC OR VDC AFTER 2 MONTHS 63 RYTN*0C'fTE5 <*W *** fftTICULflCTTES* * hHATQCST, yOL. 9 nnOGLQ01N* Gw* 9 mcv* cuetc icaons -CHd* *tC0 nicggns* *CH*iC, GW * i3 PLATELETS (X10 /m ) 33 LEuKrtcrrcs uio /** > SEUT0PHILS* * uYWPHOCYTES* 1 0ANOS* % EOSINOPHILS* % aASQPHtLS* % aONACTTES* * ATYPICAL, % nucleated BC, % PMOThOOmBIn tine* see* SOOT* lU/L SGPT, IU/L iL** PHOS.* IU/L tJtt,I*U0lN TOTAL W9 * 0ILtUI*- 0I&ECT mg l auN. mg t CHEATININt MG % LOH. IU/L ALPHA-M0OH* IU/L immunoglobulins A IU/WL G, A FACTO*, IU/WL G* S FACTO*, fU/L M. IU/WL TOTAL PWOTE1N* W 9 AL9UH1N. OM GLOtULlN. GM 1 3 ACA0P*a0E5 (NO./WM i 06ami 8,9* * ,10 U01 * 12 **.5 ; I*.A * 3 ,2 4M, l 1 .4 21.3 * .3 33*3 l 5.T ; , 6 4.* i ,6 12.0 i 1.2 86.J 1 2,1 3*3 L 0,0 i.a - 1 .* 3.3 L 0,4 0,3 i. 0.0 0*3 1 0.0 0*0 * 0,0 u.* i 104 * *3 (3) T *3*4 i 3.9 73 1 9 .4 i .0 0*3 I 0.0 13.0 * .3 a ^ 0.0 102* * 1*3 A3* * 6 10*3 * 2.1 329 * 14 4* * 9 T9 * 20 9.7 t ,1 3,3 *, I.A . .1 .2 209 i A VC 30 trm 7,03 i .12 1.92 *3.0 * 1 *.9 ,00 .4 ,0 42*3 * 1*7 21.2 i 3*.L * ,3 0 9.0 1 12.* * .3 .I*7 19,0 i 0* l 79,3 i 0*0 0,0 i, 0*0 .5 1 ,3 0,0 *. 0,0 1*3 l ,0 0*0 0*4 0.0 * 0,0 10.0 L 7* L ,1 *1' 3*.0 l 2** 00 * 0 ,1 * .1 0*0 * a.a 10.3 * ,0 *0 L o.o 5*4 227 * 9* if M* 5.7 3,* * 2*3 * ,1 .1 .2 vc 130 ?p* 0.92 * .12 1.30 * .10 *3.0 * 1**7 . .7 *2 02,2 * 1*9 21.2 * *0 3**1 * ** 0*7 * ,7 0*9 i 1*1 1**0 * 2.7 02.0 2.0 0*0 * 0,0 .9 t .5 0*0 , 0.0 J * 3 4*0 * 0,0 0*4 * 0*0 10.* * A* * * 0' 0 32.9 * 2,9 - 73 I 9 .1 i *1 0.0 * 0.0 1A.5 * uo 0 i 0.0 311 * 120 * 73 i' 20 * 5*0 * 3.9 * 2*1 1 1 .0 ,1 VC :,cqo 0.03 * *02 1*32 * *2#0 * 1*. 1 *, 02,4 * .10 *3 *2 *5 29*0 * 32*9 * *2 *A 5*9 * ,5 4,2 i ,7 9,0 * 1,1 90.3 * .9 0*0 * 0*0 .3 * *3 0,0 * 0*0 .5 * .3 0.0 i 0,4 0,0 0*4 10,4 X TO * *3 3 4/ 3*.* * l. 4. i 1 1 ,0 0*4 * 9,0 13,4 i *3 .4 * 0*0 si i 207 *. * if ay 24 * 10.2 * 2,a 290 * 34 *7 * if 47 * 17 5.5 * *2 3.5 * .0 2.0 * *2 24* i 134 VDC 53 ?P 4.57 . 1,99 i 20 .33*' *2,3 i .5*' 1*.4 L ,* 4*,9 i 1.9 22*2 i % 3*.5 : ,4 5*7 . 2 8.5 L 5 11,3 i 1.3 87,4 * 1.8 0.0 1 0.0 1 .0 *, , 7 0,0 . 0.0 0.4 L 0.0 0*0 * 0.0 0*4 * 0.0 11.3 * T* i 20.5 * 99 i .2 *' Ui s" 3 * ,1 * 15-9 i .1 41 *6 ,4 * 0.0 8*4 *, 143 342 i 47 12.2 * 310 * 1*4 12 7* i 5 103 L 2* 4,0 *, c 3*9 2,* 1 41 249 i MM tfltrlai k sub : S.E* at A rac txcivc a ttoct4 U pcnthMt< \J Slfttificaaciy 4lff*rec fro* tita Caoeral jraup, p < 0.05, ftmMCC'f wleipU o4a*Ho pea*i*t. SL 083849 34 TABLE 4 LABORATORY DATA OF FEMALE RATS AFTER EXPOSURE TO VC OR VPC FOR 2 MONTHS 43 ERYTHROCYTES txlO /hh ) 8TICUL0CYTCS* % hEHATOCRIT* vOL. \ hemoglobin* an. % -cv, CUBIC M(CQN5 HCWp, MICRO MICROOMS* MCHttCf 6* % 93 PLATELETS (Xio / ) 33 LEUKOCYTES (Xio /* NEUTROPHILS* LTMpMOCTTESt 4 4 A NQ S 1 EOSINOPHILS* 1 9ASOPH1LS* 4 hONOCtTES* 4 ATYPICAL* 1 nucleated Rac* * PROTHPOHatN Tine* SEC* SOOT* IU/L SOPT. IU/L ALU* PHOS** IU/L aiURuaiN* total g a aiLlRUatH* DIRECT HO * -fUN* HO % CREATININE* h* LQH* IU/L AL PHA --**aOM * tU/L IMMUNOOLOaULlNS A* IU/HL 3* A FACTOR* IU/hl 0. a F*CT0. IU/HL H* IU/HL total protein* ON % ALBUHIN* ON ft OLOaULlNt ON ft 3 MACRQPHAOCS (NO*/hh > 4*54 X .13 1.34 * .13 *2.3 i 5 1*.S i *2 44.2 1 22,a * 1*4 ,* 3**3 x .* 4.0 7*3 ** ,4 11.0 X 3*1 47.3 i 3*3 .3 1 .3 1.3 1 .5 0*0 * 0*0 0.0 X 0.0 o.o x 0.0 a.o i 0.0 10.4 * .1 75,0 * 10,3 32.5 i 2.4 50 i .0 * 3 *a 3.0 1 0.0 15,0 i * 3 1 .1 ITS * 4* 144 X *2 11.3 * 3.5 *4* IS 74 *. 14 51 1 15 4*0 i 3*3 * 2.4 * *2 2 .2 200 X A3 vc 30 4.41 1 *23 N+O 23 Al * 0 14.2 L ,2 43.3 * 2,1 21.4 * *4 3**1 x A 4.3 * .5 7*5 i *5 11.5 i 2*3 37.a * 2*9 0.0 i 0.0 1*0 * 0.0 0,0 i 0*0 *3 1 .4 0*0 x 0.0 0.0 1 0.0 10.4 * .3 44,3 1 9*2 35.3 i *,7 53 i 10 *1 1 0 X .1 .0 14*3 x 1,3 ,0 t .1 225 L 94 4* 1 34 4.2 * 3.4 * 2*5 1 .1 .0 ,1 7C UO spa 4.19* 1*34 * *21 ,2A *1.3 * *9 1**1 i .2 47,7 * 2.* 22.A X 33.7 5.A * .7 .2 .7 9.1 * 1.1 1 4* 3 i 5.0 44.4 i 4.A 0.0 i 0.0 1*3 X *4 0.0 ; 0.0 0.0 * 0.0 0.0 - 0.0 0.0 * 0.0 10.4 i .2 47.0 4*1 33*0 * 3*1 *3 * 4 l i .0 0,0 i 0.0 1A.0 * .9 *7 X .1 230 - 1 '5 121 * *7 4.0 i 3*4 * 2** * .2 .1 .2 luriia in maii t 3.C- ot 4 nca axe ape it oocaA la aaraBthaaaa, Slfttiflc-aaeij dlffataae ftfi m tha Caocral froup, p < 0*09* Ouaaact'a aulelpla ^oa<Mti*oa ineadun. L.OOO7C spa 4.70 * *20 1.A4 * *2.3 * 1A* 1 * .15 *4 .1 43.1 x 1*5 21.1 * .5 33.a * .3 4*1 * .3 7,2 * *9 14.4 x 7.5 02*4 ; 7,2 0.0 * 0*0 4 .5 0.0 * 0*0 3 * .3 0*0 1 0.0 0*0 * 0*0 10*4 * *2 3A.0 * 7*7 34.3 * 4*1 40 * 4 1 1 -1 *0 * .0 14.5 * .4 .4 *. . 1 521 ; ?l 204 i 14 11.1 1 2.9 453 * 35 43 i 12 53 * 15 4.1 * 3.4 * 2.5 x .2 .1 .1 190 i 1$ 'DC 53 ?p<R 4,35 x ,2'i 1.29 i 21*1 *0,5 x .4 13*9 * .2 43*4 x 22.0 x 1.2 ,4 34,5 i ,5 7.2 X . 7 4.0 x 4 14*3 X 2.9 31*4 x 2.1 0.0 i 3.0 2,0 x 1.1 0.0 X 0.0 .3 1 .3 0.0 X 0.0 a.o * a.o 10,2 * .4 73.3 x 4,J 22.4 x 2.1 3a x 2 .1 x . L .0 x .0 l 7,0 x *4 X 1.2 0.0 443 X 93 24S x 32 11./ * 1 . 7 34J x 4 A 43 X M 2* x 3 4.0 * .2 3*4 X .0 2.3 i .2 125 x 10 SL 083850 35 TABLE 5 LABORATORY DATA OF MALE RATS AFTER EXPOSURE TO VC OR VT3C FOR 3 MONTHS 43 erttnrocttcs i*ia /* aCTICULOCYftS, 4 i*T0CfltT. VOL. 1 HEMOGLOBIN* 3*1. % mCV- CUBIC MICRONS NCH4. MICRO *MCOGM$. mChnC* 5R 1 93 LATfLETS (X10 > 33 leukocytes < x10 /nr ) nEUTROPmILS* % LTnpmocYTES* 1 <UftOS % EOSINOPHILS* 4 9AS0PHILS* * *ONOCYTS, * ATYPICAL* NUCLEATED RRC, 4 pPOThPOmBIH TINE* SEC, SOOT, tu/L 5GPT* tU/L ALA, 9*05.. tu/L alLIHUBrN, TOTAL G * atLlPUilN, direct *$ * SUN* MQ % creatinine, -g a LOh* iu/l alpha-hbOh, iu/L IMHUNOOL08ULIN9 A* IU/ML 0, A FACTOR, IU/**L 9, 9 FACTOR, IU/**L M, JU/L total protein, OR * ALBUMIN* or t OLOHULIN, OR j maCPORNAOCS (NO./MR 1 Caacral 0,93 1 .33 l,3S i .Ot *3.0 i 1.3 1**6 * .A 41.9* 1 .* 31.3 l .A 3*,3 i .2 5,* 1 .5 9*3 i l.l 11.3 i *.4 55.3 i 9.1 ,3 * 3 3.3 1 .4 0,9 i 0.0 .* * .9 0.0 * 0.0 0,0 * 0.0 11.3 * .2 112 * 21 s*.a * 13.3 47 * a 0 1 0 0.0 * 0,0 13.0 * l.l .6 * 0.0 8RR * 222 37* * 41 4.0 * .3 Alt 19 At X 4 103 9 5.i * * L 3.3 * .0 2,0 l .1 130 * 94 '/C 50 ppR 4.73 *, ,07 1 ,31 1 *3,0 1 ,19 .7 l*,7 t 3 A 3.9 * .7 21,4 i .A 3*.2 i .3 4.0 1 A 8.9 i .7 13,3 1 3,1 82,8 * 3,* 0.0 * 0.0 1.3 i .9 0.0 * 0*0 .3 i ,3 0,0 c 0.0 0.0 i 0.0 11. A * .0 TO i 7 31.0 i 7a - 1.2 a .1 i .0 0.0 i 0.0 13.3 r .5 5 * ,1 544 * [A* 243 * 41 3.4 * 3.2 1 2, * 1 1 0 .0 (unua in an i J.5. of 4 rae* ottoc M asnd la / SL<niieaeLy 4tar*nc fro* t& Cdoefok itoup, 9 < 0.03, nmapec'* VC 130 pp 4.49 1.43 * 41,3 * .07 .31 .3 14.2 42,0 X 21.3 * .3 *4 .A 34.4 . 7.7 * *A .7 9,9 1 *2 11,3 2.9 87.3 X 2.1 0.0 X 0.0 1.3 * .A 0.0 3.0 .3 * .3 0.0 * 0.0 0.0 * 0.0 11.3 74 * .2 4 34.0 , 40 * 1.7 A ,1 t Q.o .0 0.0 12.5 .3 .A X ,1 903 137 232 . 44 9.7 x 3.1 X 2.4 X 1 #1 ,1 vc 1,000 PPR 4.97 1.01 *4.0 19.0 x 43.2 X 21,4 x 3**2 * .14 .23 ,A ,3 l.l .4 .3 4,5 L ,8 10,9 22,0 - 2,0 4,9 77,5 x a.a 0.9 0,0 0.0 X 0.0 x 0.0 0,0 ,5 X .5 0.0 X 0.0 0.0 X 11.0 0.0 A/ .3 " 2 X 9 33.4 X 3.5 71 x 7 .1 X .0 0.0 X t*,5 x 0.0 ,9 .5 X .1 777 * 144 335 70 9,5 *13 x ,5 A 99 X ?2 44 x 7 5,4 x ,3 3.1 * .2 2.5 * .2 2*4 * 55 eMprlaott proeadut*. /X 53 394 7.32 i .11 1.09* .0? (2) *2.0 * 1.0 (2) 1*.a *, * 3 '?i 47.3 i .5 (2) 20.0 ^ 1 1 2) 34,-1 ,2 t 21 9,2 * 3 121 8,9 * 1,7 (2) 10.5 * .5 i 2 1 89-5 1*5 iZ) 0.0 * a.a <-) .3 * 0.0 * .3 (23 0.0 (13 .3 * 0.0 * *3 (2) 0.0 (1) 0,0 * 0*0 103 *. 10 *0,0 * 51 * V0,0 43 . i i . i ( -> 0.3 i 0.0 15.0 4 13) .4 * 0,0 Oi 9*2 * 193 (3) *32 1 A* ( 3) 5.1 1 0.0 (31 2** ; 87 (3) 90 2* 131 IPN * 9 (3) 4,0 * ,2 ( 31 3.3 * .2 (3) 2.7 * L (3) 254 * 133 SL 083851 36 TABLE 6 LABORATORY DATA OF FEMALE RATS AFTER EXPOSURE TO VC OR VPC FOR 3 MONTHS 63 3TTw(0CYTtS <X10 *" l *TICLL0CtTE3* 1 **T0CffIU VOL * % *C*OGLOfn** <5M, 9 *CVw CUKtC I CRONS CHO* MtCaO NICROGHS* MCHMC* GN 1 53 OLAtELCTS (xlO /n > 33 LEUKOCYTES (Xlo /Ml* ) NEUTtfOPwlLS. % Lr*R*oCTTES. * SANOSt % eosimormili* i 9AS0*tL5* 9 'AONOCYTCS* % atypical* MUCLEATFO 0C. P*OTHflO**lN Tiwe* see. 5GOT, tU/L SGRT. IU/L alK, R03*. IU/L alLlRuSTN* Total *G * StLIflumM. OlffECT Ms \ 3UN* * CtfCATlNtftC* G 1 l0* IU/L alPha-aO* iu/l INNUNOGLOSLLINS A U/*L 3. A fACTO. IU/L Of 6 'icroff* Iu/**L 9* tu/t TOTAL *0TEIN. 3* 5 ALflU"t** G 1 9L09ULIM* 9M % 3 t*AC0R*4GC5 (NO./MV 1 CaaetaL 4*37 i .14 1.29 .24 *2.0 A U.5 i *2 46.0 * US 22.a i .4 34.6 * .2 4.2 i .9 5,a * .1 14.3 * 3.4 94.3 3.a 0.0 9.0 1.3 i .4 0.0 0.0 *3 * .3 0.0 * 0.0 0,0 * 0.0 u.o * .2 40.3 1 4,1 34.9 * 3.1 39 * * l i 0.0 * .0 0.0 13.3 i .6 .4 9.0 400 i T9 242 * 33 10.4 * . 4 424 * 4T TO i 9 79 10 4*3 .1 3.4 * *0 2*7 * .0 129 i 13 VC 50 ?P* 4.46 .13 U24 .24 41.3 * 1.3 14.2 1 .2 42*4 i 1.4 21.7 * .3 34,5 * 3 3*7 * .1 4.4 i .a 19.5 * 4.4 79,5 1 4.7 9.9 1 9.0 5 .5 o.o i 0.0 .4 * .3 0.0 1 0.0 0*0 1 0*0 10.7 *2 93,3 a 4.4 34.3 l 1.9 3* i 3 .1 1 .1 0.0 0.0 14.3 * .4 4 1 0.0 704 i 140 314 * 42 4.1 3.4 2*9 *1 .1 *1 vc 230 ppm 6.45 - . 11 1.49 - .16 42.3 * 5 14*6 .1 45.5 22.7 34.7 ,9 .3 ,2 5.9 * 4.5 * .6 .7 9.a 99.5 0*0 .3 0.0 . 1.9 1.7 0.0 .3 0.9 .3 * 0.0 . .3 o.O 0.0 11.0 . 0.0 3 41.0 * 9*1 33.0 3.5 30 4 .1 0.0 1 .3 3,9 15.3 .7 .4 0.9 421 275 * 59 19 3.9 3.3 * 2*7 * .1 .1 .0 VC 1,000 PC" 6,53 . 15 1.74 * ,06 4US ,4 14.3 .3 43.4 *4 21.9 34,4 4.2 .2 44 ,3 4.1 4 14.3 * 2.9 93,5 2.7 0,0 0.0 1.4 .5 0.0 0.0 *5 ,3 0*0 * 0,0 0.0 0,0 ll.l .1 94.3 10.5 32*3 *.9 *9 * 7 .1 .0 0,0 0.0 15.0 i ,9 .3 : .1 649 * i *4 304 * 4a 9.2 , * *53 * 19* 44 ?* 40 14 5.4 3.4 2,4 * A* * 3 .0 .3 7 *' CbetL** K4 iMB r J,;v ot * raca ** la g/ Sijaificinely dlffaraiU (th eft* Control irmi*, p < 0.05', Dwae*'a UtpU co^aciao* ?t*ca4u*a. 73C !3 3* 4*51 ** .49 a 39.3 13.5 40,3 20,7 - .07 4 *2 l ,2 ,4 34,3 * *3 5.5 .3 5.0 .4 12.a 47.0 9.0 2.* 2-7 9.0 .3 0,0 .3 O.O 0.0 0.9 9,0 0.9 o.o O.o u.o * ** 40,3 4.9 31.0 1.2 21 1 .1 9.0 9,9 1-.3 .A 4 9.0 *93 112 229 49 5.2 343 0 22 94 19 104 19 4.* *1 3.4 .1 3.0 .1 Mi 4 SL 083852 37 TABLE 7 LABORATORY DATA OF MALE RATS EXPOSED TO VC OR TOC FOR 6 MONTHS a3 SYT-0C^TS <MQ > Rff ro/lOCYTH, % *C**ATQC*IT rfOU. % g*. i *Cv* CURIC *!C*ONS -CH** nicuo mICBOG*^, 33 *WATCLEtS illO > 33 LEUKOCYTES (*10 /** > nuTAG*mIL5- * COTTEOL 6.33 .30 (?) 1**3 1 *3*5 1 13*1 ft*.4 a*.3 3**7 M3 A.A 3* (31 .A (21 . 3 12' 2* A <2) .7 (21 *3 (?) 1 i 2) .8 (2i 1**3 ,/S (3* 50 ?FM VC 8.81 1 .33 1 .12 4**8 X .5 15.9 .5 88.2 * 3.A 23.8 i 1.5 35.8 * *.7 * .7 .3 10.2 x t.o 18-3 * 2.3 :sa P?K 7C A, S9 1*30 .1* ( 11 -U *1.7 1*. 9 * ? (3i I (31 80*8 1 .9 i 31 21*7 .8 (3) 35,9 X * (31 *.3 .4 (3* 9.4 - [.-* 20.5 X 8.2 LrPHOCrTCS* % aamos* a 8*.7 1 .4 ( 3i 0*0 9.0 81.0 x 3*5 3.0 * o.o 77.3 8*0 0*0 1 9.0 e'OSlNORlLS* 9 1.0 * 1.5 i .9 2.9 - .7 4aso>***ils* * 0*0 t 4.9 o.o i o.o 0.0 - 0.9 -ONOCYTE5* * a Typical * * 0*3 L 0*0 0*0 & o.o 1*3 i .8 0*0 * 0.0 .3 t. .3 0.0 * 0.0 ^t/CLfATrO tfC# * JOOT--0Y[* TI*E* stc. AftoT. IU/L a.o o.o 12.3 *1 f 2 00.3 & 7.4 o.o 0.0 11.4 i 3 71 .0 x 7.1 0,0 10.8 49.3 0.0 . 1 Ol*' 6.3 3&PT. ru/L ala* b^rs.* ro/u 3**0 & *4 1*7 2 37,fl * 49 . 5,0 9 29,0 1.1 *3 i l hljouaih* total *<* * 0.0 0.0 0.0 * 0.0 0.0 - 0.0 aiUlHUfltN, 3[CT + 0.0 1 o.o 0.0 i 0,0 9.0 t 9.9 AU*. **<5 A 13.9 1 .4 11.3 x ,5 12.0 - *4 CPCATtMlNC* % ** X . 1 *5 * .1 4 i . 1 LQ* l u/l iu/l ln*u80GL0*t8S a* isj/hl G* a factor* tu/*L ni 1 1*2 *37 - 74 8.1 1 .5 50* 18 **1 x 158 258 1 *0 *14 i 101 2*5 - 5* G* * facto** Iu/*l 106 i 13 . Iu/"t 120 i 3 total R*orri"* v n 6.9 & 1 8*2 X .1 6*1 1 .0 ALBUWlM* G** A 3.* i *1 3-3 1 .1 3*5 i 9 GLOBULIN. GM % 3 MACROPHAGES IMO./mh ) 2.A i .7 l A* X 10* 7.9 * .0 2*8 .0 WntllS ARC IAS * STANDARD rwo* or 4 *a ccctrr as NOTED t* PARUTOrESES. / SL*niMcantLy 41<f*r*aC frn cM Control jroup, p < 0.03* Onimncs1* nuUlplo eonpnrUnn protadufA. 1,000 ?p* vc 4.7a x .22 (3) l.oo t .17 (3 > ** * 0 * 1.9 (3) 15.* * *3 (3i 88.3 X 1.7 (37 22.9 * .6 (3) 35.1 * .* I3> *.6 * 6 (3) 10.8 * 1.2 (3) 19,0 * 3.7 79,5 l 1,8 9.0 9.0 1.3 i .3 9.0 i 9.0 3 1 ,3 0.0 * 0.9 0.0 * 0.0 11.2 * .1 76,5 1 6*0 31*8 * AO * 1 ** 4 0.9 0.9 0.9 i 0,9 12*3 i *4 8 i *li' 7*0 x 21* **o i 117 7.9 * Ao *90 X 30 90 20 120 * 5 6.9 * 0.0 3.* X .0 2*6 * .0 2*8 * 81 33 FPM 7DC 7.24 m ! 1 . 79 X . 1* *5*0 1 7 15-3 t -2 82.2 X .4 21*2 i * 3**1 X .3 3.9 X .2 8.8 * ,5 12-8 X 2.* 8*. 5 X 2.9 0,9 1 0,fl 2.4 - , A 9,9 X 9.9 0.9 * 9,0 9.9 4*0 9*0 0,9 12.8 71.0 ^ 11.4 *9 X ( 5^ 9.9 x 0-0 9.9 x 0.3 12.3 X ,s . <* X * l 439 x 192 282 X 47 9.8 X l m> *?5 x 26 92 1 *6 i 6*1 i 3** X 31' .2 .1 2.7 X . 1 229 x 191 38 SL 083853 TABLE 8 LABORATORY DATA OF FEMALE RATS EXPOSED TO VC QR VPC FOR 6 MONTHS OOSE '> Pm jOYr-flQcvTe$ f*id 0 3 * CONTROL 6.*2 .21 50 TV* vc 6,65 .06 :so PPM /C 6*66 ,23 TICUlOCtTCS* ,47 *10 65 * .26 76 .25 -t*4TOC[7* vOL* * 3*. % -CV* CUBIC -tCPONS *2*3 1*.6 4l.i 1 * > .2 1 .1 41.5 . .6 15.2 *3 56,7 1 .0 *2.0 15,0 63,0 ,7 .2 2,4 *ch, *tc4o *fc*OGNS. 5 3CHP*C* G" ft *LATLET5 *MO /mm > 33 LEUKOCYTES <mo /* 21,ft i 35*3 L *.3 1 6.6 ( ,4 fA t4 21.6 36 * * *.7 4.2 .3 .i* .3 1.0 22.6 35,7 * ,5 4,1 4-6 *6 1*0 MfUTOQMtLS* % ,y0MOCYrES* 1 12*3 45.8 .3 ,6 15.5 6.6 76,6 ft.3 20.0 77.a 3*4 3,4 BANOS* * '05TM0N**n.5* ft B*5nP**[LS* * -ONOCYfES* 1 iTyfftOL. 1 nucleated brc* * 0.0 1 0.0 UK 1 .5 0.0 3.0 3 L . 1 0.0 1 0.0 0.0 1 0*0 0.0 0.3 1 ,5 .3 0.0 0.0 .3 .3 0.0 0*0 0.0 0*0 0.0 0.0 2.3 # #ft 0.0 * 0*0 o.o 4 0.0 0*0 * 0.0 o.o * 0*0 600r*ow#lN rif. SEC* LU* 1 .2 11.4 *3 11.9 2 (31 SCOT * tU/L ai .o 1 10*6 ftft.ft * 4.7 *9.4 6,6 SGPT, flj/L al*. *os* iu/l 6lLi6UN[M. TOTAL ** * 4iLlyr"* direct tui bon* i*n CEaTIM[n, <S ft l0. iu/l 4Ld4--.BD-t* tu/L r"UNOGL0HlNS A* tU/NL 37.0 6,4 22 1 3 0*0 1 0*3 0*0 1 0*0 13*3 .6 . 1 *0 952 1 71 329 45 7*7 .2 25.4 2*5 14 2 0.0 0,0 0.0 0.0 12,4 *6 .6 .1 *47 196 '*5 * 45 26*4 * 1.7 10 * 7 0.0 * 0,0 0,0 0.0 13*5 *5 * 1.6 *1 321 * 55 230 . 44 3* A ACTON* IU/"L 565 33 G* N *ACTON* lg/*L Tft 1* -* IU/nl TOTAL NWOTflN* GM ft ALU**IN. G* 1 107 6*3 3.6 3 .1 .1 7.0 4.2 .1 .1* 6.0 . 3*6 .3 m7 GLOBULIN* 3" % i maCNONmaGCS (NO./** ) 2.7 *3 .1 7 2.4 *0 2*6 .2 rtf ( wSNTUltS *R MEAJJ * STANDARD CTRO* - RATS VC**?? *S *?{ tpd pasetmes Stfnlciciotl/ dlff*rnt from ch group, r> < 0.05, Ninmct'* ailctal* cooRiftrtioa sroft.^ur*. L.COO PPM VC 6.44 ,23 (3) .65 *2,7 15.2 .16 O) .7 (3) ,3 (31 66*0 23,5 2.0 (3) -5 (31 / 35.7 *3 (31 5.2 5.7 * .6 (31 1.4 (3) 26.3 * 4.5 (3i *2.3 5*5 (3i 0*0 0.0 1.0 ,6 0.0 * 0*0 0,0 0*0 0*0 0.0 0*0 * 0,0 10,6 .0 (3) 47,3 . 2.5 35.5 1.6 *6 4 0.0 0*0 0.0 0.0 12*3 ,7 6*3 141 *.3 516 Tb 1*1 .1 69 94 .6 *1 14 1 15 6.2 5 .2 3*5 2-7 * 165 .1 .2 Y4 55 PPM vftC ft .64 .44 34.3 13*4 36*5 20.6 35.1 6.4 5*5 4 ^4 .07 .4 ,2i ,* .3 .2 .3 ,7 20.5 * 3.ft 75.4 3.7 9.0 * 0.0 *4 ,3 0*0 0.0 * 0.0 0*0 0*0 * 0.0 0.0 0.0 ll.ft *1 73.3 6.1 25.3 2.4 17 3 0.0 0.0 0.0 o.o 13.5 1*0 *6 257 [50 . ,l 67 ft* 10.4 * # 7i 669 * Vfl2 * *5 12 4,7 6.4 3.6 2*7 . 3 ,? .1 .1 101 * 34 SL 083854 39 BLE 9 LABORATORY DATA OF MALE RATS ?0SED TO VC OR VDC FOR 9 MONTHS 4 0OSE: PP" a 3 ERYTHROCYTES 1*10 /mn > RET1CUL0CYTE5. 6 HEMATOCRIT t VOL. 6 HEMOGLOBIN* 3". * mcv* cueic MfMoivs HCH9. MICRO MICROGmS. mcbc< rM * ' 53 RLATELET5 1*10 /MN 1 3J LEUaOCytES 1*10 /MX ) NEUTROPHILS. * LYMPHOCYTES* \ BANOS* * eosinophils, * SASOPhIlS* 9 ONOCTTES* 4 % NUCLEATED "PC. % R90ThR<J"9IN TIME. SEC. SCOT. IU/L SGPT. tu/L ALU. w05** IU/L BILIRUBIN. TOTAL "3 4 BIL1RUMIN, DIRECT Met BUN. MG 4 CREATININE, mg t LON* lu/L ALPHA-N90N. IU/L ImhunOGL 081NS At IU/WW G* A FACTO*. IU/XL G. B FACTO*. IU/NL ** JU/*L total ppotcin. g* * ALBUMIN. sm % GLOBULIN, GM araia xS-S mean * stwmjl. Cooeroi 50 ppa VC 7.75 3 .26 1.26 .37 *5.4 3 1.3 15.* 3 .5 7.74 * .96 * 46.0 *. 15.5 3 .27 .16 .4 .3 55.1 3 1.3 59.6 3 1.3 U ial ! 19.9 3 *.l * l0aA .4 .2 .1 .4 20.1 3 .3 33.9 * .2 4.9 * .5 19*6 * 1.2 14.5 1 1.2 74.3 3 1.3 11*3 * *.3 86*3 * 4.9 0.9 * 0.0 0.0 3 0.0 1.3 3 .3 0.0 3 0.0 2.0 * .4 0.0 0.0 0.0 0.0 11.* 1 * 103 i 6 35.5 3 3.6 4 2.0 3 1.1 9*0 * 0.0 .5 3 .3J/ 9*9 * 9.9 9*9 * 0.0 10.5 3 65 *4 u > 31.5 * *.t 44 * 4 X 0.0 9.0 J, o.o 1*.5 1.3 .7 - *0 101* ISO 62* 1 10T 9*0 1 9*9 0.0 3 0.0 15.0 3 *4 *6 i .1 323 3 179 4/ 1.3 " 196 * * & .5 4*6 * 105 113 t 29 193 * 4 6.1 3 .2 3.5 3 .1 2.4 * .2 5.6 3*4 * 2*1 * .5 .1 .5 33 * 10 jig* or root i*b sczrr as mots> n fajlwisub 230 ffm vc 7,50 * .1* 1.04 3 .12 **.5 3 1*9 1*.S * *4 59,* * 2.9 19.4 3 .4 33.* .2 4.0 6 10.9 3 .9 21.6 *.2 T*,4 3 3.6 0.0 3 0.0 2.5 3 .1 9.0 * 0.0 1.0 *4 0.0 * a*a 0.0 * 0.0 11.1 3 .1 76 * 2 37.8 * 2.4 55 3 8 1 * .0 0.9 * 0.9 16.0 * *4 .7 3 359 . 2*6 * *9 41 47 65 4/ 6*1 * 9.0 3.5 .0 2.6 1 .0 1000 ?pa 7C 7.12 3 1 a 14 * .*1 .12 *2.3 3 1.7 l*.2 .4 59.6 3 1.* 20.0 ,4 33.7 * .7 4,8 * 11.5 3 1.3 .9 2*.3 3 5.6 73.5 3 6.2 .3 3 .3 1.3 * .9 0.0 3 0.0 .8 * .3 0.0 3 0.9 0.0 3 o.o 11*4 .3 81 * 11 *3.3 3 9.4 *7 * 9 9.9 * 0,0 0.0 3 0.0 15.5 3 .3 .6 3 *1* * 248 ; .1 93 4/ 52 4/ 5.4 *3 476 * 112 123 * J6 66 * 17 6*0 * .1 3.* 3 .1 2.6 * .2 49 3 2* */ SlfaiEleaotly ilEfarwe fta* tha eaaenl (rasp, * < 0.05. Otmaee'i wlelpla ea^aruon ptocadara. 55 ppa VDC 6.53 1 j/ *16 1.39 I .36 *2.5 * ,4 14*3 * *5.1 3 ,1 1-1J/ 21.9 3 -*J/ 33.B * *2 4,Y 9.5 26.3 i 6.7 72.0 i 6.* 9*0 1 9.9 US 3 1.0 0.0 3 .3 . 0.0 .1J/ 0.0 - 0.0 3 ^ 12.0 87 3 2 I3l4/ 21.3 * 37 * 3.3 (3) 7 i i| 0*9 * 0*9 9,0 * 0.9 12.0 3 1-2 13) , r i .Of? * 5*7 214 * 66 3 51 * 2 13) 7 niJ/ 6.0 ; .1 f1> 3.A * 0.0 (3t 2,6 1 .1 (31 99 i 14 SL 083855 40 TABLE 10 LABORATORY DATA 07 FEMALE RATS EXPOSED TO VC OR VDC FOR 9 MONTHS ertthrocttes <*10 heticulocttes. t hEHatOCBIT. VOL, 1 hEHOGLORIN. G**. % CV, CUBIC "ICBONS cna. ico hicrqgms. mchbc. i S3 platelets ixio /mm 1 33 LEUKOCYTES 1X10 /mm ) vEuTWOPMILS. * LThphoctTES. SAMOS, S EOSINOPHILS. * BASOPHILS. MONOCYTES, * ATYPICAL, NUCLEATEO BBC, PROTHROMBIN TIME, sec. SGGT, IU/L SGPT. IU/L ALK. PH(1S., IU/L siLisuPtN, total h a bilirubin. direct -o* SUN, MG t CREATININE, HO * LU IU/L ALPHA-HBOH, IU/L IhhunOGLOBINS A, [U/HL Ot A FACTOR, IU/ML o, a factor, iu/hl H, Itl/HL total protein, on % ALSUMIM. OH t GLOBULIN, OH S haCBOPhaSES (NO./hh | 6,*2 , .16 (3) 1.06 * .3* *1.7 * .7 (31 14.1 m.m 61.1 * .1 (31 *3 (3) 20.7 .4 (31 33.9 * .3 (3) 3.3 * .3 (31 5*2 * . 1 (3) 15.0 * U9 32.3 0.0 2.0 0.0 l.s . .6 0.0 * 0.0 .3 * .3 0.0 * 0.0 0,0 * 0.0 11,0 .4 (3> 74,3 * 3.3 30,3 * 2.3 26 * 3 .0 4 .0 0.0 0.0 17*0 * 1.6 ,7 * .0 602 * 102 374 * 62 6,3 * 1.2 560 150 34 12 34 * 24 4,4 . .2 3.7 * 1 2.7 53 .1 4 6.ST * .41 1.0T * 10 AO.O *4 13.3 * S*,4 SI ,3 * .3 4.2 1.2 33.* * .5 3.A .5 3.3 .7 ST.3 5.7 70.3 * 5.A 0.0 * 0.0 .3 * .3 0.0 0.0 2.0 * .7 a.o 0.0 0.0 0.0 n.i * .2 00.3 . 4.3 3*. 3 * 5.5 ST * 5 0.0 * 0.0 0,0 * I*.3 * .T * *., * S99 * 0.0 .0 .0 46 27 3.1 * 3.A * S.* * .3 *3 .3 2TCBIZS All >AN 3 STANDARD SUUA 07 FOUR RAIS EXCEPT AS NOTED Cl PAUSHUSES. 0.76 .67 in I .60 * .27 (3) *0.0 * 1.7 (31 13.6 * .6 (3> 59.6 * 2.7 131 SO.3 * Ua (31 3*.l ** .3 (31 *.a *4P 3.6 * .3 (31 *6 (31 SO.3 * 5.9 (3) 76.7 5,4 (3) 0.0 * 0.0 1.3 * 0.0 *m 1.0 ,4 0.0 .4 0.0 * 0.0 0.0 * 0.0 11.0 # .2 (3) 7*.3 14,9 S9.S * 4.4 ss 0 .1 * .1 0,0 . 0.0 16.0 . 1.7 .7 . .0 332 * 131 229 * 84 6.2 *.0 * s.s * .2 .2 .1 6*40 * 34 <31 94 ,19 (3) 40.7 . 1.2 (3 14,1 * .* (31 49.9 ** 1.3 (31 ?o, a * .* (31 34.a * .1 (3) 3.3 * .3 (3) s,a * 1.1 (3) 28.3 . 70.3 * *.l (31 * .6 ( 3) 0.9 . .4 * 0,0 * 0.0 .s o.o .5 * .3 0,0 0.0 0.0 . o.o 11.1 .1 (3) 93.3 11.9 (3> 42,0 * ?6 . 0.0 * *.* <3> 3 (31 0.0 0.0 0.0 19.7 m S.9 (3) .a .1 (31 444 I6S (1) 312 # 97 13> 5.6 .0 (3> 378 * 139 9 * 16 (31 69 * 1* (31 6.0 . .2 (3) 3. a 2.2 .1 (3) 3 (3i 44 36 6*0 - U 14 39.5 13.4 41 ,6 20.9 14,0 `.31 1* * l. 5.3 <6 . . * 17,6 . 7, 80.0 0,0 US 0.0 .8 . 2. 0. * 0. 0.0 0.0 . 11.4 * 0, 0. 72.a * 24.3 , 17 7, 1. 2 0.0 0, 0.0 3, 0. t**fl ml * 444 21* *20 - 133 5.9 . 450 * *7 94 * 19 61 * 3 4.* 4.1 2*3 51 13 SL 083856 41 TABLE U T.ASORATORT DATA. CY "ALE PATS EXPOSED 70 vc a* <nc rot U MOTTO 005E PR* E^ythhocytes t*10 /MW ) RETICULOCYTES* S HEMATOCRIT. VOL. * hWOOL09IN, OH. 1 HCV. CUBIC MICRONS HCH9. MICRO Ml GROOMS. *CH$C * GM * 3 PUATELCTS <A10 /MM ) 33 LEUKOCYTES <X10 /MM ) NEUTROPHILS. * LYMPHOCYTES* * 3ANOS % EOSINOPHILS. * 3AS0PHILS. * MONOCYTES' * ATYPICAL. NUCLEATED RBC, * PROTHROMBIN TIME. SEC. SGOT. IU/L SORT. IU/L ALK. Rmos.. IU/L 9ILI RUB IN. TOTAL mb BILIRUBIN. DIRECT MOB 9UN, MG i CREATININE, mg LOH. IU/L ALPH4~HBDH* IU/L IMMUNOGLOeiNS 4. IU/ML control 0.94 .20 1.03 * .11 *5.0 u.s * B .2 6*.a i t.l 21.4 * .5 33.0 * .4 4.2 t 10.2 t .A .6 27.3 L 2.4 TO.4 2.4 0.0 * 0.0 l.S t .5 a.o i 0.0 -9 * 4 0.0 1 0.0 0.0 1 0.0 11.2 i .1 SB 12 34.* L 1.7 SI t 4 a.a * 0.0 o.o * 0.0 IS.* I .0 *6 t .0 SOB L 108 302 L 61 7.2 * *6 30 pjn VC 4.SO * .19 1 . *9 .*4 *2.9 * 1*4 1*.2 1 .4 66*0 i 2*3 21.A i .B 33.1 i .5 5.4 * .4 IU* * .5 22.1 1 *.T 74.3 5.0 0.0 i 0*0 .9 * *3 0*0 * 0.0 .A .4 0.0 0.0 0.0 0.3 10.S 1 .* (7) *5 t 3 24.9 * 2.7 4* * 4 .0 * .0 0.0 0.0 15.* L .B 4 * .0 13B 1 SlA/ RT 17i/ ISO PRO '/c 7.S2 t .24 .78 * **.l 1 14.6 1 .B .3 SB.9 *. 1.2 19.8 i .4 33.7 * .2 5.7 * .4 i. ; .4 2U8 * 3.9 77.0 i 3.8 0.0 * 0.0 6 i .2 0.0 * 0.0 .6 * .4 0.0 0.0 0.0 0.0 10.9 . .3 S3 l 3 24.4 1.4 39 - 3 #0 * .0 .0 * .0 15.4 .5 #6 * i 224 i 4ai/ 148 38 a* a factor, iu/ml 331 * 17 s. a factor, iu/ml 101 4 M. IU/ML TOTAL PROTEIN. ON ALBUMIN. Gm % SLOWULTN. GM 1 104 L 21 5.9 t .1 3.4 * .1 2.2 * .1 3.8 3.3 i 2.3 .1 .1 .1 4.1 * 3.7 2.3 t .2 .2 .2 MACR0RMAGC5 (NO./MM ) 64 * 30 E8TRIZS ARE MSA STASDAID IRXOR OF limit RATS UNLESS OTHERWISE INDICATB) 01 PARENTHESES. a; S'.pitflcnc'-v 4l!f*ruc ir<* ttM control (roup, p < 0.03, DunaAtt' mulctpU ctpMtt*on proemdut*. i.coo spa vc 4.32 i ,*9 l.*8 1 .*6 41. S 1 2.0 13.7 *6 67,2 * 3.0 22.2 * 1,2 33.0 S.7 .5 #4 24,S 16.5 24.5 3.3 42.3 l 8.3 9*4 * 9*4 .9 * .2 0*0 * 0.0 .4 * 0.0 * .2 0.0 *3 i. .5 10.s * .4 134 i 94 45.3 ` 21.5 91 - 37 ,0 * .0 0.0 i 0.0 L A , 0 * 1.1 *5 .1 343 * 64 218 L 3.7 * 3B .3*-/ 264 64 92 l 4 *3 *, 20 5.7 .1 3.4 i .1 2.3 i .1 3 r 44 A 53 7.72 i .83 i *4,8 * .u ,4 1*,6 * 58.1 * -2 .a 19.0 * ,2 32.7 L 5.0 1 -Z .2 8.9 * .4 15.3 1 2.1 82.6 1 2.2 0,0 i o*o 1.3 * .6 0.0 1 0.0 *4 i .2 Q.O * 0,0 0.0 L 0.0 11.3 * .2 59 1 35.1 3 .3 46 1 0.0 * 0.0 0.0 * 13.* * 0,0 97 .6 i 366 * .i 74 230 * 93 4.5 4*1 40 98 * 36 * 6.0 * ,4 3.9 i *1 2.2 i .1 50 i 25 42 SL 083857 TASL1 12 LUCTATWT DATA OT FEMALE RATS ERYTmROCYTES 1X10 /MM 1 RETICULOCYTES * HEMATOCRIT* VOL. A HEMOGLOBIN* Gm. a MCV. CUBIC MICRONS 5.26 .41 (7) 1.56 i .19 *2.3 1 1.0 t 7) 13.9 1 ,4 (7) 63.7 7.4 (7) 6,99 1.16 43,6 14,3 62,4 *19 .14 .9 3 1*3 5.50 L 6.56 c 35.1 * .91 { 7) 4.27 <7? 4.6 (7) 11.7 * 1*6 (7) 69.1 6.0 (7) *.S3 - 3.33 17.00 - 16.J3-" 33.5 - 9.S 10.A - 3.6 31.1 - 18.9 6.90 * 9 iJ 43*4 13.9 61*4 .05 - 1? *3 -1 *5 MCHB. micro MICROSMS. MCHBCf gw * 53 PLATELETS < *10 /mm ) 33 leukocytes (XLO /mm > 27.5 2.4 (7) 32.a .3 t 7) 5.6 6*9 *6 (7) 1.0 (7) 20.4 32.9 5.0 5.a 4 *3 *5 .5 23.6 * 33.9 * *. a * 10.6 * 1.5 (TO *6 (7) .7 (6) 4,0 (7 J 35.1 A.A 31 .A 1.9 6,3 .7 30.3 11.7V 20.1 * 32.7 4.3 4.1 .3 *1 *3 neutrophils. * 37,3 z. 3,4 20.6 4.4 35.9 * 6*5 (71 A3.0 - 16.0 21.0 2-7 LYMPHOCYTES. A 70*a 1 3.6 79*0 4.4 63.3 * 6.4 (7) 55.0 15.0 79.0 2*7 BANOS. A 0,0 0.0 0,0 0.0 .1 . ,1 o.o 0.0 0.0 0.0 EOSINOPHILS. 1 1.9 6 *4 2 .5 . .2 3.0 2.0 .6 *2 BASOPHILS. A monocytes, a atypical, a nucleated rbc. a PROThhqnRIN time. Stc. SSUT. IU/L SSPT. IU/L ALK. PHOS.t iu/l 0*0 1 0.0 1*0 1 *3 0.0 1 0.0 0,0 0,0 10.9 .2 43 5 34*5 2.0 34 3 0.0 0,0 .4 *4 0.0 0.0 0,0 0.0 10.6 *2 41 3 34.4 1*9 19 3 0.0 * 0.0 .1 * .1 0.0 0,0 0.0 * 0*0 11.3 * .9 (7) 71 . 22 3S.6 * 10.2 36 . 7 0.0 1.0 ~ 0.0 0.0 1.0 0.0 0.0 0.0 12.0 1.2 ANT a3<>V SI.5 60. ^ 70 49 V 0.0 * 0,0 .4 0*0 * 0*0 0.0 * 0.0 10,5 *3 61 6 39.9 i 3.3 24 * 3 BILIRUBIN. TOTAL mb 1 .1 .0 .0 *0 .3 * .2 .1 .0 0 -0 9ILI RUB IN. OIRECT Md* BUN. HO A 0,0 0,0 16,9 3.2 0.0 0.0 16,4 1.1 .1 30.6 *1 5,0 0.0 I5.S 0.0 .5 0.0 * 0*0 14*5 CREATININE, mo A LON* IU/L .7 .0 414 103 , 6 .0 473 96 .7 * .0 791 * 355 .7 0.0 1099 913 6 351 12* alpha-mboh, iu/L ImmijNOGLOBINS A. Iu/ML 3. A FACTOR. IU/ML 345 53 6.3 1 *6 439 30 294 46 ABO * 160 730 616 5.8 .5 303 336 219 4,4 * 73 1 * 4-- 304 2 3. 8 FACTOR, IU/ML 46 3 54 38 p*a 13 ( M. IU/ML 91 17 70 3a 54 12 TOTAL PROTEIN. SM % 6.6 .2 6*4 * *2 6.6 6.A .A 6*5 1 alBumin. GM 1 3LOBULIN. GM A 3 MACROPHaSES (NO./mh i 4*0 3*7 * 31 * *1 *1 17 4*0 * 2*4 * *1 *3 3.R * 3.* * .2 *2 3.7 3.7 * A03 ,2 .3 4.2 2.3 34 *1 *1 ID ENTRIES AM MAN STANDAM IRSCR OF SICHT RAIS UNLESS OTHESWTSE INDICATED IN PARENTHESES. THE VALUES FOE THE 1,000 PW VC CROUP AM THE MEAN 5.2. OF TVC RAIS. 1/ Siiaiilcaady dfTaranc from eha coacxal group, P <0.03, Dunnacc'a nulcipLa cOBoartaoa procadura. 43 SL 083858 TABLE 13 NUMBERICAL DISTRIBUTION OF CHROMOSOMES IN BONE MARROW OF RATS EXPOSED TO VC OR VDC treatment Control VC (1,000 ppm) VDC (55 ppm) Control VC (1,000 ppm) VDC (55 ppm) Control VC (1,000 ppm) VDC (55 ppm) Control VC (1,000 ppm) VDC (55 ppm) VC (1,000 ppm) VDC (55 ppm) No. of Chromosome Frequency Rats * 40 41 42 43 * 44 Tetraploids (4N) oer 100 cells After One Month b/ 6 3^/ 4 42 1 0 0.25 0.11 6 4 4 41 1 0 0.33 0.17 6 3 3 43 1 0 0.42 -- 0.08 After Three Months 5 6 7 34 2 1 0.0 0.0 5 9 7 30 3 1 0.0 0.0 3 5 5 39 1 0 0.0 0.0 After Six Months 5 5 6 36 2 1 0.2 0.2 5 3 6 35 1 0 0.2 0.2 6 6 5 36 1 0 0.33 ,k 0.21 After Nine Months 5 7 3 36 1 1 0.4 i. 0.25 6 6 5 37 1 1 0.5 0.22 6 4 6 38 2 0 0.5 0.22 After Twelve Months 4 3 3 41 2 1 0.25 0.31 2 6/ 6 38 2 0 0.5 0.86 a/ Mean b/ Mean S.E. c/ Chromosome frequency based on numerical counts of 25 metaphase spreads. 44 SL 083859 TABLE 14 MORPHOLOGICAL ABERRATIONS Of CHROHOSOMJ.S IN BnM: IlliUh'-'A' OF RATS EXPOSIT* TO VC OR VDC Treatment Ho. of Mice Chromatid Capa per 50 Celia Chroma tid Breaks per 50 Cells TranslocaLiors pet 50 C..Mis total Aberrations per 30 Cells Control VC (1,000 ppm) VDC (55 ppm) 6 6 6 After One Month 0.67 0.21--^ 0.83 0.31 0.33 0.21 0.33 0.33 0.50 0.22 0.87 0.48 After Three Months 0.33 0.21 0.33 0.21 0.17 0.17 1.33 0.21 1.67 0.56 1.37 0.61 Ol Control 5 0.80 0.40 VC (1,000 ppm) 5 0.14 0.14 VDC (55 ppm) 3 0.33 it 0.33 0.0 0.0 0.0 i 0.0 0.53 * 0,53 After Six MonLha 0.25 0.25 0.20 -e 0.20 0.0 0.0 0.05 * 0.32 0.34 * 0.21 0.86 0.47 Control VC (1,000 ppm) VDC (55 ppm) 5 5 6 0.66 0.27 0.20 0.20 0.60 0.28 0.26 0.26 0.0 0.0 0.33 0.21 0.20 0.20 0.0 0.0 0.0 0.0 0.12 0.52 0.20 0.20 0.93 0.33 SL 083860 (Continued to next page) TABLE 14 (Concluded) irea rntent No. of Mice Chromatid Chromatid Wreaks per 50 Celts Translocations per 50 Cells Total Aberrations per 50 Cells Control VC (1,000 ppm) VDC (55 ppm) 5 6 6 After Nine Months 0.0 0.0 0.33 0.21 0.33 0.21 0.0 * 0.0 0.17 0.17 0.0 0.0 0.0 0.0 0.17 0.17 0.0 0.0 0.0 0.0 0.67 0.33 0.33 0.21 After Twelve Months o> VC (1,000 ppm) 4 0.0 0.0 VDC (55 ppm) 2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 a/ Mean S.E. b/ Chromosomal aberrations based on analysis of 25 metaphase spreads. Co r< o *98q TABLE 15 CQT.TAGEK CONTENT IN LIVER AND LUNG 0? RATS EXPOSED TO VC OR VDC FOR 6 MONTHS Liver Controls Vinyl Chloride (1,000 ppm) Vinylidene Chloride (55 ppm) Lung Controls Vinyl Chloride (1,000 ppm) Vinylidene Chloride (55 ppm) Collagen Content Males (w/wO_____ Females 188.9 , 25.4/ 259.8 42.1 254.7 44.0 226.0 72.2 213.0 16.6 213.0 16.6 329.8 111.3 545.9 97.9 612.3 165.6 H o 363.6 166.2 364.2 26.5 425.6 a/ Mean S.E. of four rats. SL 083862 47 TABLE 16 COLLAGEN CONTENT IN LIVER AMD LUNG OF RATS EXPOSED TO VC OR VDC FOR 9 MONTHS Collagen Concent (ug/gm) Males Females Liver Controls 283.8ll45.6^/ 206.2181.1 Vinyl Chloride (1,000 ppm) 264.0i30.2 256.7156.4 Vinylidene Chloride (55 ppm) 330.7187.8 349.21143.7 Lung Controls 151.3135.3 Vinyl Chloride (1,000 ppm) 225.9126.9 Vinylidene Chloride (55 ppm) 298.6147.5 a/ Mean 1 S.E. of four rats. 188.4131.0 178.0129.3 273.01107.9 48 SL 083863 TABLE 17 COLLAGEN CONTENT IN LIVER AND LUNG OF RATS EXPOS FT) TO VC OR VDC FOR 12 MONTHS Liver Controls Vinyl Chloride (1,000 ppm) Vinylidene Chloride (55 ppm) Lung Controls Vinyl Chloride (1,000 ppm) Vinylidene Chloride (55 ppm) Collagen Content Males (uz/m) Females 425.6 i 42.6^/ 383.4 106;2 335.0 73.4^ 293.0 i 45.4/ 465.6 128.6 331.6 52.6 256.7 32.5 266.2 139.7 151.6 57.^ 303.2 117.4^ 320.2 136.2 294.4 55.1 a/ Mean = S.E of four rats, b/ Mean S.E of three rats, c/ Mean S.E of two rats. 49 SL 083864 Terminal Body Height Brain Kidney Spleen Testes Liver Kidney Spleen Testes TABLE 18 ORGAN WEIGHTS OP MALE RATS AFTER EXPOSURE TO VC OR VPC FOR i MONTH Control VC 50 pm VC 250 ppm VC 1,000 ppm VDC 55 ppm Absolute Height ftml 350 452/ 2.11 0.03 10.4 0.9 2.91 0.13 0.75 0,06 3.35 0.05 4.9 0.4 1.4 0.1 0.36 0.03 1.59 0.02 371 t 27 1.82 t 0.25 11.4 : 0.9 3.36 t 0.40 0.73 t 0.05 2.98 b 0.14 367 11 1.98 0.02 11.2 0.5 2.87 0.08 0.64 0.02 3.07 0.00 384 26 2.04 0.04 12.4 0.7 3.20 0.13 0.80 0.02 3.18 0.11 Relative to Brain Weight {Ri/.Riy 6.6 t 1.0 2.1 b 0.7 0.43 t 0.08 1.77 t 0.34 5.7 0.3 1.5 0.1 0.33 0.01 1.55 0.02 6.1 0.2 1.6 0.0 0.39 0.00 1.56 0.05 363 3 1.87 0.04 11.0 0.5 2.98 0.03 0.64 0.04 3.12 0.10 5.9 0.3 1.6 0.0 0.34 0.02 1.66 0.03 a/ Mean S. E, of four rata. co tr> o 03 Ui CD TABLE 19 ORGAN WEIGHTS OF FEMALE RATS AFTER EXPOSURE TO VC OR VDC FOR 1 MOUTH Control VC 50 ppib VC 250 ppm VC 1,000 ppm VDC 55 ppm Absolute Weight fgml Terminal Body Weight 235 A 15s7 235 8 230 8 210 A 8 230 X 1 Brain 1.91 0.06 1.95 0.04 1.88 x 0.04 1.81 A 0.07 1.74 X 0. 05 Liver 1H/1 Kidney Spleen 6.9 X 0.2 1.82 0.09 0.46 0.02 7.3 A 0.2 1.92 A 0.06 0.51 A 0.03 7.5 x 0.2 1.88 x 0.02 0.48 x 0.02 7.5 0.3 1.77 X 0.10 0.48 0.04 7.0 X 0. 2 1.55 0. 23 0.50 0. 04 Ovaries 0.22 0.06 0.13 0.01 0.14 0.01 0.17 0.03 0.12 X 0. 02 Liver Kidney Spleen Ovaries 3.63 A 0.05 0.96 x 0.06 0.24 0.01 0.12 x 0.04 Relative to Brain Weight (em/finO 3.72 0.09 0.98 A 0.02 0.26 A 0.02 0.07 0.01 4.01 x 0.15 1.00 x 0.03 0.26 x o.Ol 0.07 x 0.01 4.19 0.28 0.98 0.06 0.27 0.02 0.09 0.02 4.05 0.,22 0.90 X 0.,14 0.29 A 0.,03 0.07 X 0.,01 SL 083866 a/ Mean S.E. of four rats. TABLE 20 ORCAN WEIGHTS OF MALE RATS AFTER EXPOSURE TO VC OR VDC FOR 2 MONTHS Control VC 50 ppm VC 250 ppm VC 1.000 ppm VDC 55 ppm Absolute Weight (gw) Terminal Body Weight 477 9a/ 475 11 451 16 488 15 479 11 Brain 2.14 0.05 2.09 0.04 2.19 0.02 2.21 0.01 1.70 0.28 t: 12.1 0.4 12.4 0.7 13.0 0.8 14.6 0.7 12.7 0.6 Kidney 3.32 0.17 3.25 0.08 3.30 0.08 3.65 0.13 3.13 0.11 Spleen 0.81 0.05 0.74 0.05 0.76 0.08 0.83 0.08 0.79 0.05 UK)l Testes 2.83 0.27 3.27 0.16 3.53 0.17 3.32 0.10 3.08 0.06 Relative to Brain Weight fgm/gm) Liver Kidney Spleen Testes 5.7 0.3 1.55 0.09 0.38 0.02 1.3 0.1 6.0 0.4 1.55 0.03 0.35 0.03 1.6 0.1 6.0 0.4 1.51 0.03 0.35 0.03 1.6 + 0.1 6.6 0.3 1.65 0.06 0.38 0.01 1.5 0.1 8.4 1.9 2.07 0.46 0.51 0.09 2.0 0.5 a/ Mean S.E. of four rats ir* o /LS)8 8 i j ii i TABLE 2L ORGAN WEIGHTS OF FEMALE RATS AFTER EXPOSURE TO VC OR VOC FOR 2 MONTHS Control VC 50 ppm VC 250 ppm VC 1.000 ppm VDC 55 ppm Absolute Weiftht (anQ Terminal Body Weight 271 A ga/ 274 A 9 273 * 12 265 A 3 270 A 11 Brain 2.08 A 0.04 2.16 A 0.09 2.03 a 0.03 1.98 A 0.05 1.84 A 0.03^ Liver 7.A 0.3 8.0 A 0.8 8.1 A 0.2 7.9 A 0.1 6.9 X 0.3 Kidney 1.97 0.06 2.13 0.07 2.02 A 0.03 1.95 0.05 1.59 X 0.18 Spleen iun* Ovaries 0.59 0.05 0.14 A 0.02 0.54 A 0.05 0.18 A 0.02 0.58 A 0.04 0.15 0.00 0.50 0.02 0.13 X 0.01 0.48 X 0.01 0.11 A 0.00 Liver Kidney Spleen Ovaries 3.57 A 0.16 0.95 A 0.01 0*28 A 0.03 0.07 A 0.01 Relative to Brain Welftht ({WesI 3.66 A 0.20 0.99 A 0.03 0.25 A 0.03 0.08 A 0.01 4.02 A 0.09 1.00 A 0.02 0.29 * 0.02 0.07 * 0.00 3.98 a. 0.06 0.99 A 0.04 0.25 X 0.01 0,06 .L 0.00 3.75 A 0.20 0.86 A 0.11 0.26 U_ 0.01 0.06 0.00 a/ Mean S.E. of four rata. b/ Significantly different from control, p < 0*05, Dunnett's Multiple Comparison procedure. SL 083868 TABLE 22 ORGAN WEIGHTS OF MALE RATS AFTER EXPOSURE TO VC OR VDC FOR 3 MONTHS Control VC 50 ppm VC 250 ppm VC 1.000 ppm VDC 55 ppm Absolute Weight (gm) Terminal Body Weight 554 2i/ Brain 2.22 0.05 Liver 15.0 0.4 Kidney 3.65 + 0.19 Spleen 0.76 + 0.03 U1 Testes 3.4 + 0.1 Liver Kidney Spleen Testes 6,8 0.3 1.65 0.12 0.34 t 0.02 1.53 0.06 519 35 2.18 0.05 13.8 0.5 3.28 0.18 0.80 0.12 3.5 0.1 516 2.22 12.8 3.42 0.78 3.4 37 0.07 1.3(3) 0.10(3) 0.07(3) 0.0 500 12 2.23 0.04 14.2 0.6 3.30 0.21 0.83 0.07 2.9 0.5 Relative 6.3 0.1 1.50 0.06 0.36 0.05 1.61 0.04 Brain Weight (gm/gm) 5.6 1.49 0.34 1.54 t 0.6(3) 0.03(3) 0.03(3) 0.06 6.4 1.48 0.38 1.29 0.3 0.10 0.03 0.21 520 17 2.36 0.22 13.9 0.3 3.43 0.33 0.77 0.07 3.4 0.2 6.0 0.4 1.51 0.24 0.33 0.02 1.50 0.17 a/ Mean S.E. of four rata except as noted in parentheses. SL 083869 t ,i i i ' .1 I TABLE 23 ORLAN WEIGHTS OF FEMALE RAT'S AFTER EXPOSURE TO VC OR VDC FOR 3 MONTHS Control Terminal Body Height Brain Liver Kidney Spleen Ovaries 322 lO3^ 2.06 0.08 8.6 0.4 2.08 0.06 0.55 0.04 0.14 0.01 Liver Kidney Spleen Ovaries 4.16 0.16 1.01 0.03 0.27 0.02 0.07 0.00 VC 50 Plan VC 250 ppm VC 1.000 ppm VDC 55 ppm Absolute Height Cgml 309 6 2.13 8.4 0.2 2.26 0.05 0.63 0.06 0.17 0.02 o uj 284 2^ 2.05 0.01 8.4 0.3 2.22 0.07 0.58 :L 0.03 0.16 0.01 305 10 2.02 0.02 8.7 0.5 2.13 0.08 0.57 0.02 0.15 0.01 278 9^ ---- 1.91 0.03 7.1 0.3 1.98 0.04 0.48 0.02 0.17 i 0.02 Relative to Brain Weight (Rm/Km) 3.97 0.09 1.06 x 0.01 0.29 0.03 0.08 0.01 4.08 0.16 1.09 0.04 0.29 0.02 0.08 0.01 4.29 i 0.27 1.06 0.04 0.28 0.01 0.08 0.01 3.74 0.18 1.04 0.02 0.25 0.02 0.09 0.01 a/ Mean S.E. of four rats. b/ Significantly different from control, p < 0.05, Dunnett's Multiple Comparison procedure. SL 083870 TABLE 24 ORGAN WEIGHTS OF HALE RATS EXPOSED TO VC OR VDC FOR 6 MONTHS Group Control Ab 50 ppm VC 250 ppm VC 1,000 ppm VC 55 ppm VDC Terminal body weight Brain Liver Kidney Spleen Testes 632 + 302/ 660 + 12 2.82 + 0.18 2.41 + 0.11 16.6 + 1.9 16.4 + 0.7 4.13 + 0.33 4.09 + 0.20 1.17 + 0.05 1.10 + 0.06 3.29 + 0.32 3.40 + 0.14 686 20 2.17 + 0.13 17.4 + 0.6 4.01 + 0.19 0.95 + 0.11 3.59 + 0.07 607 + 0.32 2.11 + 0.08 16.4 + 1.4 3.74 + 0.24 0.86 + 0.03^ 3.46 + 0.20 623 + 15 2.17 + 0.06 16.0 + 1.0 3.87 + 0.20 0.85 + 0.03k/ 3.49 + 0.24 Relative to Brain Height (gm/gm) Ul Liver O' Kidney Spleen Testes 7.6 + 1.6 1.88 + 0.30 0.53 + 0.06 1.50 + 0.25 6.8 + 0.1 1.70 + 0.06 0.46 + 0.04 1.42 + 0.10 8,1 + 0.8 1.87 + 0.15 0.45 + 0.07 1.67 0.09 7.4 + 0.6 1.69 + 0.09 0.39 + 0.02 1.57 + 0.12 7.4 + 0.4 1.78 + 0.10 0.39 + 0.01 1.61 + 0.10 a/ Mean + S.E. of four rats. b/ Significantly different from Control group (Dunnett's multiple comparison procedure). to TA.8C80 SL 083872 i i L .. J I I1 i. i TABLE 25 ORGAN WEIGHTS OP FEMALE RATS EXPOSED TO VC OK VDC FOR 6 MONTHS Group Control 50 ppm VC Absolute Weight (gm) 250 ppm VC 1.000 ppm VC 55 ppm VDC Te rmina1 body weight Brain Liver Kidney Spleen Ovaries t 360 + 29^ 315 + 34 2.16 + 0.03 2.08 + 0.08 8.7 + 0.7 8.7 + 0.3 2.16 + 0.11 2.38 + 0.17 0.61 + 0.04 0.58 + 0.06 0.30 + 0.13 0.13 + 0.03 329 + 12 2.13 + 0.06 8.8 + 0.3 2.31 + 0.15 0.69 + 0.08 0.19 + 0.02 310 + 11 2.06 + 0.07 8.7 + 0.3 2.08 + 0.14 0.07 -06 0.21 + 0.03 347 + 16 1.87 + 0.0^ 8.4 + 0.4 2.14 i 0.16 0.53 + 0.04 0.15 + 0.02 Relative to Brain Weight (Rni/gm) Liver Kidney Spleen Ovaries 4.05 + 0.35 1.00 + 0.06 0.28 + 0.02 0.14 + 0.06 4.21 + 0.02 1.15 + 0.08 0.28 + 0.02 0.06 + 0.01 4.16 + 0.14 1.09 + 0.07 0.32 + 0.03 0.09 + 0.01 4.21 0.22 1.01 + 0.07 0.34 + 0.04 0.10 + 0.02 4.48 + 0.18 1.14 0.05 0.28 0.02 0.08 0.01 a/ Mean + S.E. of four rata. b/ Significantly different from Control group (Dunnett's multiple comparison procedure). Il TABLE 26 ORGAN WEIGHTS OF MALE RATS EXPOSED TO VC OR TOC FOR 9 MONTHS Control 50 PPM VC 250 PPM VC 1.000 PPM VC 55 PPM VDC Absolute Weight (gm) Terminal Body Weight Brain Liver Kidney Spleen Testes 682 + 4C/ 2.37 + 0.06 13.0 0.9 4.10 0.12 0.98 + 0.03 4.43 + 0.79 669 + 18 2.27 + 0.11 16.7 + 0.9 4.11 0.25 0.93 0.06 3.63 0.14 724 + 22 2.38 + 0.03 18.7 + 0.5 4.27 + 0.11 1.07 + 0.05 3.82 + 0.13 658 + 15 2.47 0.08 17.9 0.9 3.90 0.15 0.92 -r 0.11 3.58 -Im 0.29 688 + 27 2.13 0.07 14.9 + 0.9 4.07 + 0.19 0.88 + 0.06 3.37 0.12 Relative to Brain Weizht (zm/zm) Liver Kidney Spleen Testes 7.6 -L 0.4 1.73 + 0.05 0.41 + 0.01 1.86 0.30 7.4 + 0.4 1.81 + 0.12 0.41 + 0.04 1.61 + 0.12 7.9 + 0.3 1.80 + 0.05 0.45 + 0.02 1.61 + 0.06 7.2 + 0.4 1.58 + 0.08 . 0.37 + 0.05 1.46 + 0.14 7.0 + 0.4 1.91 i 0.08 0.41 _ 0.03 1.59 + 0.10 a/ Mean + . E. of four rats. 58 SL 083873 TABLE 27 ORGAN WEIGHTS OF FEMALE :*ATS EXPOSED TO VC OR VTC FOR 9 MONTHS Control 50 P'?M VC 250 PPM VC 1.000 PPM VC Absolute Weight (gm) 55 PPM VC Terminal Body Weight Brain Liver Kidney Spleen Ovaries 383 + 37--^ 2.17 + 0.06 12.0 + 2.0 2.36 + 0.09 0.59 0.04 0.10 + 0.03 307 + 36(3) 2.14 + 0.10(3) 7.9 + 1.2(3) 2.18 + 0.31(3) 0.55 + 0.05(3) 0.14 + 0.03(3) 347 + 25 2.21 + 0.10 10.9 + 0.9 2.51 + 0.16 0.61 + 0.10 0.14 + 0.01 341 + 15 2.12 + 0.08 10.5 + 0.3 2.46 + 0.06 0.55 + 0.09 0.11 + 0.03 Re la five to Brain Weizht (zm/zm) Liver Kidney Spleen Ovaries 5.6 + 1.0 1.09 + 0.05 0.27 + 0.01 0.05 + 0.02 3.7 + 0.6(3) 1.02 + 0.16(3) 0.26 + 0.03(3) 0.07 ^+w 0.01(3) 5.0 + 0.5 1.14 + 0.07 0.28 + 0.05 0.06 + 0.01 5.0 + 0.2 1.16 + 0.03 0.25 + 0.04 0.05 + 0.01 a/ Mean + S. E. or four rats excep t as noted in parentheses. 404 + 24 1.97 + 0.04 10.2 + 0.9 2.63 + 0.12 0.51 + 0.01 0.15 + 0.01 5.2 + 0.5 1.34 + 0.06 0.26 + 0.01 0.08 + 0.00 SL 083874 59 1 TABLE 28 ORGAN WEIGHTS OF MALE RATS EXPOSED TO VC OR VPC FOR 12 MONTHS Control 50 ppm VC Absolute Weight (gm) 250 ppm VC 1,000 ppm VC 55 ppm VPC Terminal Body Weight Brain Liver Kidney Spleen Testes 815 is*/ 2.31 0.04 18.9 0.3 4.43 0.13 0.9 0.1 3.67 0.16 824 24 2.33 0.04 22.3 t 1.4 5.07 0.29 1.1 0.0 3.57 0.06 792 16 2.40 0.04 19.6 1.0 4.48 0.27 1.0 0.1 3.40 0.13 722 40 2.33 0.05 20.5 + 1.5 4.33 0.19 2.3 1.3 3.52 0.19 769 + 30 2.21 0.05 17.3 0.8 4.40 0.09 1.0 i 0.1 3.61 0.09 Relative to Brain Weight (gm/srm) Liver Kidney Spleen Testes 8.2 + 0.2 1.92 0.07 0.4 0.0 1.60 0.08 9.6 0.5 2.18 0.12 0.5 0.0 1.54 0.05 8.2 0.4 1.37 0.11 0.4 0.0 1.42 0.05 8.8 0.7 1.86 0.09 1.0 0.6 1.52 0.09 7.9 0.4 1.99 0.04 0.4 0.0 1.64 0.06 a/ Mean S.E. of eight rats. 60 SL 0838^ i i.i .i ,i ji >i t> i t L-i I TABLE 29 ORGAN WEIGHTS OF FEMALE RATS EXPOSED TO VC OR VDC FOR 12 MONTHS Control 50 ppm VC Absolute Weight (gm)______________________ 250 ppra VC 1,000 ppm VC 55 ppm VDC Terminal Body Weight Brain Liver Kidney Spleen Ovary--------------- 473 15^ 2.18 0.04 11.2 0.6 2.85 0.19 0.7 0.1 416 15 2.14 t 0.02 10.4 0.4 2.50 0.10 0.7 0.0 391 24--^ 2.13 0.02 14.2 2.4 2.51 0.09 1.2 0.5 412 62 (2) 2.15 0.07 (2) 19.9 7.6- (2) 3.11 0.00 (2) 2.1 1.1 (2) 0.12 -* 0.03 (7) 428 22 2.08 0.05 10.6 0.4 2.70 0.07 0.6 0.0 0.17 1 o.or Liver Kidney Spleen Ovary 5.1 0.3 1.31 0.09 0.33 0.03 0.06 + 0.01 Relative to Brain Weight (gm/gm) 4.9 0.2 1.17 0.04 0.33 0.02 0.08 + 0.01 6.7 l.l 1.18 0.05 0.57 0.21 0.08 + 0.01 9.2 3.2--^ (2) 1.45 0.05 (2) 0.94 0.50 (2) 0.06 + 0.12 (2) 5.1 0.2 1.30 0.02 0.29 0.02 0.08 + 0.01 a/ Mean iS.E. of eight rats unless otherwise indicated in parentheses b/ Significantly different from control group, p < 0.05; Dunnett's multiple comparison procedure. SL 083876 TABLE 30 SUMMARY OF TUMORS THAT OCCURRED IN RATS DURING 7 TO 9 MONTHS EXPOSURE TO VC OR VDG^7 250 PPM VC Sex Female Male Rat No. 197 200 252--; Tumor--/ Week Terminated 39 39 39 1.000 PPM VC Female 269 270 271 288? 39 39 39 33 287--/ 35 286SJ 38 55 PPM VDC Male Female 305 360^ 39 39 Liver n a io s arcoma 33 Lung Hemangiosarcoma 3 Mammary Gland Duct adenocarcinoma 2 Skin (subcutaneous) Hemangiosarcoma Squamous cell carcinoma 3 Adrenal Gland Sebaceous gland Adenocarcinoma 11 3 1 13 3 2 1 a/ Includes only rats which were found to have tumors during 7 to 9 months expc^Ae to VC or VDC. b/ Severity of lesion: 1 - mild; 2 - moderate; 3 - marked; 4 - severe, c/ Died or unscheduled termination. No tumors were found in control rats during this period. 62 SL 83877 TAUl.g 31 a/ SUHHAHY Of HJHOBS 11IAT Oa.UftRKO 1H UUHTttOL HATS AND HATS EXPOSE!) IQ IQ 12 HOffl'IIS 10 SO OK 250 ffH VC~ Tuiaork/ Cuntrot 50 omu VC 250 ppm VC Sex Rat Ho. female Male Female Ma le Female 41 94 I44t/ 143t/ 165 166 167 179-^ 201 202 203 205 207 20# 2t.lt/ 2l2t/ 2 lit/ 210t/ 209t/ Week Terminated 52 52 40 50 52 52 52 50 52 52 52 52 52 52 48 49 49 51 51 Lung Nemangloaarcoma 2 Squaaioua cell carcinoma 1 Liver IIcutang 1 oearcona 3 1 23 1 33 Hepatoma 1 He wary gland AdeiHKArclnow Fibroadenoma 22 22 2 Spleen Malignant lymphoma 2 Omentum llema ng 1 oa a rcoma 2 Mesentery Hemangloaa rcoma 4 Skin IIeiaang 1oaarcoma 3 Pituitary Chromophobe cell adenoma 2 3 33 1 32 at Includes only cat a that were found to have tumora during 10 to 12 montha exposure to VO. b/ Severity of lesianat l - slid; 2 - Moderate; 3 - marked; 4 - aevere. c/ Death or unaclteduled termination. SL 083878 TAM.E 12 SlttHAKY Of mums Wl Ot-'l.-IHlKHi IH RATS WHIlNi: III til 12 HIHTTIlS EXIYISUHi: 1 I .Wilt rm V UK 11 I'm <4T>* j/ Includea only rata that war* (oarf to tiava tuaore during IU to 12 ewiitlie uapoaure to UC or VDU. b/ Severity ol leatonai I - alld; I - wtderace; J - marked; k - aevure; I - queel tollable, c / Death or unaiiieduled tcrnlaatton. t-i 6 i*tBO ................................................................................ TABLE 33 J CUMULATIVE TUMOR INCIDENCE IH RATS DURING 12 MOUTHS EXPOSURE TO VC OR VDC Control I 50 ppm VC 1 250 ppm VC i 1,000 ppm VC 55 ppm VDC Tumor Lung Hemanglosarcoma Bronchiolar adenoma Squamous cell carcinoma 0/35 0/35 0/35 0/35 0/35 0/35 0/36 0/36 0/36 0/36 0/36 0/36 0/36 0/36 1/36 3/34 0/34 0/34 4/34 1/34 0/34 9/36 0/36 0/36 0/36 0/36 0/36 0/35 0/ 35 0/35 Liver Hemangloaarcoma Reticuloendothelial cell carcinoma Hepatoma 0/35 0/35 0/35 0/35 0/36 0/36 2/36 10/34 0/35 0/35 0/36 1/36 0/36 0/36 0/36 1/36 0/34 0/34 6/34 15/36 0/34 0/34 1/36 0/36 0/36 0/35 0/36 0/36 0/35 0/35 Mammary gland Due t adenocarclnoma Fibroadenoma Malignant lymphoma 0/35 0/35 0/35 1/35 0/35 0/36 0/36 0/36 0/36 0/36 0/36 0/35 0/36 0/36 0/36 5/34 1/34 1/34 0/34 0/36 0/34 " 1/36 1/34 2/36 0/36 0/36 1/35 0/35 0/36 0/35 Kidney Adenoma 0/35 0/35 0/36 0/36 0/36 0/34 1/34 1/36 0/36 0/35 Hemangloaarcoma Omentum Mesentery Mesenteric lymph nodes Skin 0/35 0/35 0/35 0/35 0/35 0/35 0/35 0/35 0/36 0/36 0/36 0/36 0/36 0/36 0/36 1/36 1/36 1/36 0/36 0/36 0/34 0/34 0/34 0/34 1/34 0/34 0/34 0/34 1/36 3/36 0/36 0/36 0/36 0/36 1/36 1/36 0/35 0/35 0/35 0/35 Adrenal Hemangioma 0/35 0/35 0/36 0/36 0/36 0/34 0/34 2/36 0/36 0/35 SL 083880 (continued to next page) TAIiLE 33 (Concluded) CUMULATIVE TUMOR INCIDENCE IN RATS DURING 12 MONTHS EXPOSURE TO VC OR VDC Control 50 own VC 1 250 ppm VC 1.000 ppm VC Skin Squamous cell carcinoma Keratoacanthoma Fibroma 0/35 0/35 0/35 0/35 0/35 0/35 0/36 0/36 0/36 0/36 0/36 0/36 0/36 0/36 0/36 0/34 0/34 0/34 1/34 1/34 1/34 0/36 1/36 0/36 Sebaceous gland Adenocarcinoma 0/35 0/35 0/36 0/36 0/36 0/34 0/34 1/36 Pituitary Chromophobe cell adenoma 0/35 0/35 0/36 0/36 0/36 1/34 0/34 0/36 55 ppm VDC of i 0/36 0/36 0/36 0/35 0/35 0/35 0/36 0/35 0/36 0/35 of Humber of rats with tumor/totaL number of rata that died or were terminated during 12 months expoeure to VC or VDC. SL 083881 IJ J i "1 \ TABLE 34 SUMMARY OF TISSUE LESIONS AND M/E RATIOS IN RATS AFTER EXPOSURE TO VC OR VDC FOR 1 MONTH gj Severity of lesion] 1 - 114, 3 - Moderate, 3 - earlied, 4 - severe, $ - questtollable, IlM ' not available m x? o CtPP CCPO to TABLE 35 SUMMARY OF TISSUES LESIONS AND M/E RATIOS IN RATS AFTER EXPOSURE TO VC OR VDC FOR 2 MONTHS ! |/ Severity of lesion; 1 - alld, 2 - aoderete* 3 * narked, 4 - severe, questionable CO IT* O CD U> 00 CbDJ * TABLE 36 SUMMARY OF TISSUE LESIOHS AND H/E RATIOS IN RATS AFTER EXPOSURE TO VC OR VDC FOR 3 MONTHS i ik i SL 083884 a/ Severity of lealuna: 1 - mild, 2 - Moderate, 3 - marked, 4 - eevere, J - questionable TABLE 37 SUMMARY OF TISSUE LESIONS AND M/E RATIOS IN RATS EXPOSED TO VC OR VDC FOR 6 MONTHS Ualontr^ Sex: Rat Wo,; LrOntl OU Malea Feinu 1 ei 11 ii 14 49 50 51 Via \ 4 ,WU |f at. --a _______ t,,u Ha le Fema t es Males 152 219 130 231 232 265 266 26 268 301 302 303 304 337 338 339 340 Eye Heart Hyocardltla till 1 1 1 2 2 11 1 l 1 Lung Chronic aairine pneiaaonla l Bapli||(c i'oata cell Infiltration A focua of papillary prollfera- l i 1111 1111 2 1 1 1 l t1 1 1 11 11 1 1 l.lvar Micro foci of Mononuclear cel la i t t i 1111 1111 11 1111 till Portal Inflaanatlon O i il11 11 1l1 1 l1 1 1113 1111 Pancreaa Chronic Interetltlal pan- Cecum i 1 1 1 Kidney Chronic lnteratltlal nephrltla MlcrocaIculL _ PyeUt U_______________________________ Teat la Tubular degeneration 1 necroala tilth Minerallaa- i i i1 l 3 1l1 11 1 1 I 1111 1 1 1 111 I Braln Hemorrhage Bone Marrow U/JC Ratio 11 1 Li. LA _1~3_ LA U5_ 1.A _l-5_ LA L2_ LA _l-J_ 1.3 _l-l_ 1.2 _U_ 1.2 .U4_ 1.1 _L4_ l.A _LJ_ 1.A 1-3 Ttauue* not llated were normal. aj Severity of lealona; 1 - lld, 1 - Moderate, 3 - marked. 4 - aevere + - questionable. SL 083885 TABLE 38 SUMMARY OF TISSUE LESIONS (OTHER THAH TUMORS) AND M/E RATIOS It) RATS EXPOSED TO VC OR VDC FOR 9 MONTHS--^ SL 083886 tj Do*a uot lac I death* aad iiMclitdulad UralMtloni that occurred 4url*| / t 1 enitthi. y Severity of Ualonei 1 - alld; I - noderate; ) - Marked; h - aavera; - fueetlouable. K^41 cn TAB I K 1 gWMA AY OF T1 SSK LESKMS (WHEN HIAll TWHNS) _ AHH H/K KATIOS IN Mil 6* l'< 5* H> TO VC ON VI)C Fl II HnNTHSg/ Him mil I .QUO |Pn (VC) Hule* Eenuilej Ha lea fenalaa__ in ii m21 21 21 21 21 2k 21 2* 51 5! S Sfi ii i2 il ii ?37 m 23S 2A0 241 24? 243 244 *> Hetloal degauaratlux l 1I _ Ursilla_____________________________________________ 1------------------------------------------ Heart _ tlydcirillili___________________ 1---- *----1---- 1--- 1---------------- *- Luii| 0__ ipdU i(lQMBeUMnia- __ _____________________ l_____ _________I_____i________ i--------- Liver |>t a tended aluuaotda ftbroala Hlcrofocl of monouuc tear cal la Itltllll Vacuollent Lon (nodular nr dlaaetainated) _ Orjenaratl loti and/or nacroala _1___________ _________ __________________ ______ i iiii ii Cvcua _ fljiwariv.lA iuaeji___________-____________ Kidney HlcrocalculI Chronic luteratitlal naplirltla I 1I I Tubular dilation and caata PlfcMCut In tuhnlea _ fyfilflnephrlti*_____________________________ 1Ur i tvary ladder _ iidiiMitiiim_________________________________ Taatla _ lybol** UK.cnexatlft^-ftd_nftciuila Adrenal lienor rhah* Dlacaadad alwuaolda 1 _ iteaoiUoiU -_______________________________________________________________________________________________ pancreaa _ IulojiU Jiauciutitia____________________________________________________________ __________ ________________ Spleen Extramedullary InaatopoUilt _ pfirg | ftaia_______________________________________________________________________________________ tone Harrow Smear _ U/J (_____________________1*-- 1-1 -J-A- i-Jk -M- 1* 1-tt i*.5_ 1*1 i*i i*a u M -2*0- l l 2*1 i*_ 15 23 3 IA 083887 t* TAtil.e )9 (CiNicludoil) .leaI.ona jug 310 3M 312 Jgg. U*\*L 3U 116 ms HI HI Hi H2 Hi HI Heart I _ Choglc gutta*JW!igl_____________________________I___ 1____ Liver Hicrofocl of mononuclear cell* Vacuullaatloa ( ulnUr or I 111 1)21121 dtiaa(M(4) Dciteueratio* iM/or Mcroala I _ f*f{l_Ll inl**Ia iac4l,i,l______________________________ CeciMa _Ptmwmi_ln Xu5eGt ________________ _________ ____ ________1 Kidney Clironlc lateral Itlal Mphrltli I _ dlU|.lgRiftd_caaUav _ Pituitary Cyat____ Adrenal _ DUtju^ed * I[maoIda Paacreca v| Chroute |iaacreat l^t^a_ Buna Harrow Saeer _ H/f facto____ 1.4 _|i6_ LI LB X-i I S Lli*_ 1! I 1.7 1.6 I, I. */ Doe not Include uaachtduled terminal Iona tliat occurred duriu* Ift to 12 nonlba. hi Stv.rUy of Ualooai I - alnlMli 1 - od*ra(o; 1 - aevero; 4 - vary tavero; aid j - 4.1*11 imalila. SL 083888 TABLE 40 NUMBER OF pFATMS AND UNSCHEDULED. TERM!NATIONS .OF,_MIC repos ro to VC OR vtC Trmatment Control VC-50 ppm VC-250 ppm VC-1,000 ppm VC-55 pprn Sex Male Female Male Female Male Female Male Female Male Female During Exposure Month 6 7 3 9 jO 11 12 - - 1 1 m * *m 1 1 111- 1 1 244 21 - 142l- 1 3 4 10 * 2353m- - 29 64 *> mm - - - 2* - - ----- 1 74 SI 0s3889 i i LJ ) 1 TABLE 41 LABORATORY DATA OP MALE MICE AFTER EXPOSURE TO VC OR Vf>C FOR 1 MONTI] Vinl EHYlHIiOCYIES 63 (KIO /HH 1 HET1CULOCYIES. ft IlfHATOCHIT* VOL. ft IIEHOGL Oil IN. GH. ft HCV CUIt 1C HI CHUNS MC1 ill HI CKO MlCHUGHS. HCIIHC. GH ft 53 PLATELETS 1X10 /HH 3 t J LEUKOCYTES 1X10 /HH 1 NLUTHOPHILS* ft LYHPHOCYTES. ft HANOS* ft EOSINOPHILS* ft 11AS0PH 1L S * ft HONOCYTESt ft ATYPICAL* ft NUCLEATEO HOC * ft SGPT. IU/L HUN. HO ft HACHOPHAGIS 3 INO./HH 1 Cun trot 8.17 .66 l.ofl 4_ 45.5 4_ .09 .6 13. A .3 56.8 A.h 16.7 1.2 29.4 .5 3.6 _ 1.0 A.2 .S io.o 4 A.3 80.0 5.1 0.0 * 0.0 1.0 * .6 0.0 0.0 1 .0 4_ .6 0.0 4 0.0 0.0 * 0.0 5TO <2) 21.5 * 2 (I) 20 * 5 (21 VC 50 ppM 6.33 4_ .80 (3) 1 .no *_ .28 Ol 44.3 ?.T (3) 13.0 4. 11.3 4_ 1.2 4.9 (3) <3)S/ 20.T 4_ 1 .0 (1) 29.? 4_ 1.0 O) 6.1 .9 (3) 5.? * .6 (3) 27.0 4.H 71.3 4_ 4.2 0.0 0.0 1.3 .5 0.0 0.0 .5 .5 0.0 0.0 .3 .3 1711 X 59 28.5 X 9 (21 (2) VC 2 SO Hnu 6.99 .48 1 .63 .28 45.H * 2.8 15.1 1 .4 65.5 21.8 1.7 1. 5^ 33.4 *_ 2.6 4.7 1.1 6.3 *_ .8 15.8 * 2. 1 83.3 2.0 0.0 0.0 .3 * .3 0.0 0.0 .8 4_ .5 0.0 _ 0.0 0.0 * 0.0 119 A S (2) VC 1,(100 ppm 7.27 4 .43 .74 4_ .23 47.3 4_ 2.1 14.0 4 .6 65.2 4_ 1.7 19.3 29.6 4 .5 .2 4.6 * .9 6.4 4_ 1 .2 18.11 *_ 4.5 79.3 * 4.8 0.0 0.0 .8 .5 0.0 0.0 1.0 1.0 0.0 0.0 0.0 0.0 95 i 2 (21 1 7.5 X l (2) 11 _ " (1) SL 083890 entries are aiean S.g. of 4 alee except aa noted In parentheses, a/ Significantly different frua the Control group, p 0.05. Ikmnutt's Multiple coaparlaon procedure. vw: Si ppm 6.8 3 i. 1.2 7 *_ .44 .21 4 6.5 4_ 1.2 14.0 * .6 68.6 4 2.9 20.7 4 l.U 30. 1 .7 5.4 4_ .8 6.8 4_ 1. 1 9.5 4 2. 1 89.8 t 1.8 0.0 * 0.0 .5 4 .3 U.0 4 0.0 .3 .3 0.0 4_ 0.0 0.0 4^ 0.0 282 - 19 (2) 19. n J 3 <21 72 4_ 24 TABLE 42 IABORATORY DATA OF FEMALE MICE AFTER EXPOSURE TO VC OR VDC FOR 1 MONTH 63 (UYlHHOCYItS 4X10 /MM ) HtncuLOcyits, HEMAIOCRil. VOt. * HEHOOLOHIN. GM. ft HCV. CUIIIC M1 CHUMS HCItll, MICRO HICROtiHS . HCIIUCt 6H % 53 PLAtFtflS 1X10 /MM 1 33 LEUKOCYTES IK10 /MM 1 nluthophils. ft LYMPHOCYTES. HANDS, ft EOSINOPHILS. ft RASOPHILS, ft MONOCYIfS, ft ATYPICAL, ft NULL LA ILO HOC. ft SGPf. 1U/L IIUN t Kb ft 4 MACHOPHAfitS INO./MM 1 Control 7.*7 .14 .96 49.0 .10 .9 14.S .4 65.6 l.U 19.5 .5 29.6 .6 5.5 .a 9.0 2.1 12. J 2.0 04.11 * 2.a 0.0 L 1.5 4 0.0 .6 0.0 4. 0.0 l.s *_ .6 0.0 4_ 0.0 0.0 0.0 171 i 47 (2) 22.0 i. l CD 71 t. 21 (3) VC so piH 6.9H bit 1.19 * .31 4 7 .5 * 2.7 14.6 .B 6U.9 4_ 3.2 21.2 4^ 30.0 4^ 1.0 .2 6.3 4_ .7 9.1 4_ 2.0 15.3 *_ 2.3 At .3 3.4 0.0 4 0.0 3.3 1 .4 0.0 4 0.0 .3 4^ .3 0.0 4 0.0 0.0 4 0.0 197 1 0 (D 21.4 - o.s (D to tr< Entries are naan 1 li.E. ot 4 ttlte accept aa noted In parenOieeea. VC ISO ppu 7.21 .29 1 .60 4 .34 4B.3 . 14.5 2.4 .7 66.9 * 1.0 20.2 4^ .4 30.1 4^ .3 5.0 4 1.3 (3) B.9 4 2.3 13.11 4_ 3.1 H4.0 4_ 4.0 0.0 . 0.0 2.0 .A 0.0 0.0 ,5 4^ 0.0 4_ .3 0.0 0.0 4_ 0.0 143 * 84 VC 1,000 ppu 7.16 1 .33 (3) 1.06 t . 14 O) 44./ 2.2 (3) 12.9 * .7 (3) 62.4 i 2.0 (3) 17.9 28.6 * .2 (3) .9 (3) 4.1 .9 (3) 3.6 i .3 (3) 24.7 * 5.6 <3> 7 3.0 5.0 (3) 0.0 0.0 <3) 1.3 * .0 (3) 0.0 1 0.0 (1) .5 1 .6 13) 0.0 t 0.0 (3) 0.0 * 0.0 13) 201 (l) 19.0 d) 20 i 5 <D voc SS ppn 7.40 .66 (J) 1.71 . SH 4 | . * 7.4 16.1) 4 .6 <3> 66.6 4 2. 3 (3> 20.4 4 30.7 4 H o> .i (3) 3. 7 x .4 <D 5.3 4 l .4 (3) 15.1 4 4 . A 3 3 6 6.5 0.0 * 0.0 1.0 4 .6 O.U .5 t 0.0 .3 0.0 O.U 0.0 * 0.0 311 J 40 24.0 A 0.0 ID <D 20 - s <D 1* 083891 SL 083892 TABLE 43 LABORATORY DATA OF MALE MICE AETER EXPOSURE TO VC OR VDC FOR 2 MONTHS Conetol VC SO ppia VC iSO pp*U VC ltOOO ppia 63 EHV TllllOCVIf S 1*10 /MM 1 s.he i icni ncrn * iichatdcrii. vm . * ilfHIKil IHIIM. AM. * HCVt CUBIC Ml CHONS HCHII HICHO HICHDGMS. MCIIIIC. AM f 53 PL AllLfIS I X1 A /MM ) 33 LEUKflCVICS 1X10 /MM 1 -I-^1 nfiithhphil s. % |VMPHAC.VlfS. % HANDS * EOSINOPHILS. % HASOPIIILS. * HONOCVIES. * AfVPICAL. * NHCLEATEO HHC. * SAP I. 1 ll/L HUN* HA % .1 MACHOPIIAAES INO./MM 1 A. A A * .17 I.IB .14 411.A 1.3 14.2 .5 75.0 1.2 22.1 .4 29.5 .2 5.a A 5.9 .0 40. ft 1 1.0 57.S 1 1 .9 A.A t A.O .3 .1 A.O * 0.0 1 .5 1.0 0.0 0.0 0.0 # 0.0 129 40 35.5 2.0 23 * 11 A.00 .22 1.70 * .07 4S.B #_ 1.3 13.A 4 .3 76.3 * .7 22.7 .4 29.11 .3 6.3 4_ .7 4.4 .5 43.0 7.9 54.3 7.1 0.0 0.0 1.0 *_ .6 tl 0.0 4_ 0.0 .3 .3 0.0 * 0.0 0.0 0.0 H7 24.0 12 3 4--^ 6.7H 4 .H8 * 50.0 .26 (1) .11 (1) 1.0 (3) 14 .H 4 .5 (1) 73.9 * 2.0 (3) 21. H 29.S * . 1 (3) .7 <31 5.9 * .7 6.H .7 (1) JU.I * 13.6 60.5 1 14.2 .3 .1 .5 * .3 0.0 0.0 .5 * .1 0.0 0.0 .3 .3 104 4_ 14 23.0 1.7* 6.S9 .19 1.10 * .30 40.B 4 .6 14.1 *_ .2 74.0 1.3 21.7 .3 29.3 .1 6.0 1.0 6.1 4_ .9 39.0 1 1. I 59.5 10.7 0.0 0.0 1.0 .4 0.0 4_ 0.0 .5 4_ .5 0.0 0.0 0.0 * 0.0 163 * 22.S * 162 * 35 1 ?A^ H2 <3) Cdtrlea are Mean * S.E. of 4 alee except aa noted In parentheaea. a/ Significantly different trow the Control group, p <-0.04, IJunuett'a Multiple coaiparlaon procedure. vim: SS P|MH 6. 17 * . 19 1.31 .11 45.3 * .5 13.5 * .1 71.2 *_ i.a 21.1 4. 29.9 .6 .2 4.0 .4 4.4 * 1.4 31.0 4 4.4 60.5 4_ 4.1 0.0 # 0.0 0.0 0.0 0.0 0.0 .5 .5 0.0 0.0 0.0 4 0.0 216 4 24 IS . 0 1.9 07 4 50 TABLE 44 LABORATORY DATA OF FEMALE MICE AFTER EXPOSURE TO VC OR VDC FOR 2 MONTHS 6 .1 EHVIHHUCY It S 1X10 /MH 1 HtTICULOCYltS. % litMAIOCWIT* VOL. * UlMOlil.OH 1N OH. * HCV. CUHIC MICRONS HCMIH MICRO MICROHMS. HCHHC. r.M % 53 PLATLLEIS 1X10 /MM 1 33 LtUKOCYTES I X10 /MH > NIUTHOPHILS. * LYMPHOCYIES. HANUS * % CUSINOPHILS. * HASOPMILS. * HOMOCVIES. * A 1YP1CAL % HUCLEAIFO ROC. * SGP1. IU/L HUN, MG % .1 MACHOPMAGLS (HO./MM ) Control 5.94 1 .18 .18 .07 46.8 3.3 13.8 t 1.0 78.7 * 1.8 23.2 1 .5 29.5 i .2 5.1 .0 5.0 t .3 29.3 i 6.8 67.5 i 5.9 0.0 * 0.0 2.5 i .9 0.0 i 0.0 .8 1 .5 0.0 * 0.0 0.0 1 0.0 119 1 18 23.3 *. .9 213 * '23 <3> vc 50 ppia 7.09 * .24 1.13 * 54.0 * .17 1.7 15.9 .5 76.2 .5 22.5 .1 29.5 .1 4.1 .6 5.3 * .3 26.H * 7.3 71.11 7.3 0.0 0.0 1.0 .6 0.0 *_ 0.0 1.7 .4 0.0 * 0.0 0.0 * 0.0 200 1 25 24 3 1.3 vc 250 ppm 6.52 .10 1 .04 50.5 .23 l .2 15.1 78.0 * .1 2.8 23.4 1.0 30.0 A .6 4.5 * .3 5.0 .5 22.5 H.6 76.5 8.6 0.0 * 0.0 .0 .5 0.0 0.0 .3 .1 0.0 0.0 0.0 0.0 296 * 027.5 _ 7? 2.3 vc l.UUO ppia 6.66 *- .30 1.26 * .24 49.5 * 1.7 14.6 .5 74.5 2.1 21.9 * .7 29.5 .7 5.7 .3 5.1 .5 29.0 12.0 68.8 4 11.3 0.0 0.0 1 .8 1.1 0.0 0.0 .5 .3 0.0 0.0 0.0 0.0 222 40 24.8 * 1.0 III 6 entries are wean t S.E. of 4 nice except as noted In pnrentlieeee, ./ significantly different fro the Control group, p < O.OS, Ikinnell*e Multiple coapariaon procedure. vue 55 6.90 * 1 .5U t .41 .07--/ 50.8 1 .9 15.2 .5 73.8 1.7 22. 1 * 29.9 * .6 .2 4.9 * .5 4.8 * .2 22.8 6.2 75.8 5.6 0.0 * 0.0 .5 * .5 (1.0 1.0 4 0.0 .6 0.0 9 0.0 .3 * .1 217 t 31.3 4 SO I.is' 21 H t Utt ( SL 083893 ji j TABLE 45 LABORATORY DATA OF MALE MICE AFTER EXPOSURE TO VC OR VPC FOR 3 MONTHS b3 IHVlHHOCVUS 4X10 /HH 1 HI 1 ICULOLmS, % iitHAiucinr. vol. * mmX.LOHlH. OH. * MCV t f.UHlC Hi CHUNS MCIIU, HICHO MICHOt.MS. HCHIICt GM t S3 I'LAlIrLflS 1X10 /HM t 3J vO LtlJMlCYItS (XIO 2HH 1 NtUIHUHHILS. * L YHHIKJCVlt S. % HAHObt % tOSlUUIXIllS, % H ASOtJlt 1L S * MONItCmb. % AIVt'lLAL. % NUCl L AUD HOC. * SSHI, IU/L HUN nr. * J| HALrtttl'HAlit S INO./MH 1 Control 7.52 4_ l.ll 4_ .Ah o> .33 (1) AS.3 4_ .9 (3) 1S.S 4 .A (3) 64 ,b 1 20.7 3. A .a (3) <3> 32.1 4 .7 <3> 7.0 4^ 1 .2 <3> b.2 4 1.0 (3) 22.3 4_ 3.3 77.0 4 2.9 0.0 0.0 .s 4 .1 0.0 4_ 0.0 .3 4^ .1 0.0 4_ 0.0 o.o 4^ 0.0 S5.0 4^ 9. 1 20.0 1 .1 106 4 I2l. VC "ill ppia H.1S t ,2 1.23 1 .2 49.0 lb. 3 .A SO. ft 19.9 * .1 14. | (?) in U> (2) (H (7) (1) b.2 (1) 7. 1 1 .0 (7) 2S.lt 12. b 73.H 4_ 12.7 0.0 4 0.0 .3 4 .) 0.0 4_ 0.0 1 t .1 0.0 4_ 0.0 0.0 4_ 0.0 9b. 7 21.s (3) 19.0 1.2 (1) VC p|M* 0.01 t i (2) 1.63 t A (7) Ab.O 12 0 (7) IS. 1 * 7 (7) S7.S tl (71 19.1 * 3 (7) 33.3 1 (7) 3.9 il 0 <7> 30.S 4 til.a 4 .S 0.9 1.0 1 .0 0.0 0.0 0.0 4 0.0 .a 4_ .0 0.0 0.0 2.0 4 2.0 72.0 4_ b.9 24. a 4_ 1. 1 Entries are mean + S.E. of 4 mice except as noted In parentheses. VC 1 ,1)00 ppw b. | 3 4 .tifi 1 .S3 4 . 2S 4b. 3 4_ 13.3 4_ 1.2 1.3 (31 bft.l 21.9 4 .9 (31 .7 31.3 4 b.n 4_ ,s (3) .9 (J> 3.H 4_ .7 is.3 4_ 2.1 H4.3 4 2.1 0.0 4 0.0 0.0 4 0.0 0.0 4 0.0 .S 4_ .s 0.0 4_ o.o 0.0 *_ o.o H6.0 4_ 9.2 (3) 21.0 4 2. 1 (31 S3 4 10 vnc 5i pin 7.14 4 .22 1.24 4 .20 44.0 4 .9 14.S .4 SI .7 4 1.2 20.3 4 .1 3 3.0 4 .S 7. 1 1 .7 b. 1 4 1 .2 19.0 * H.h HO. 11 4 U.S 0.0 4 0.0 0.0 4 0.0 0.0 * 0.0 .3 4 .1 0.0 4 0.0 0.0 * 0.0 112. S 4 S.9 27.0 4 2.4 t IS 0 99 SL 083894 TABLE 46 LABORATORY DATA OP FEMALE MICE AFTER EXPOSURE TO VC OR VDC FOR 3 MONTHS hJ t hy ittiiacvits mu /MM 1 Ht T l CUt tlC Y 11 S * * IILHA10CHI I VOL. Y> UtMOfil OHIO. 6H. X MCV. COMIC MlCrtOMb HCIIU. M1CMO HlCttOliMS. MCHHC. OH S 5J ptAffLtrs mo /mm i OoO 1J LIOKOCYIIS CKlQ /MH 1 NtUlHOHIilLS. % lvhhhocvtfs. * HANDS* * tOSINOPIIILS. HASOPHILS. * MUNOL'VUS* * AlYflCAL* * NOLLt MM HOC. * SOI* 1 . 1 U/L HUN. HO * 1 MACHIlPtlAl't S INO./HM 1 Control 6.73 * 1.0* * *4.3 14.1 * .17 (1) .07 (3> 1.2 (3) .5 (3> 65.9 4_ .3 (3) 20,9 .2 o> 31 .H .2 (3) 5.9 .5 (7) 3.2 4. .* (7) 12.0 t 3.2 HT.3 4_ 3.6 0.0 *_ 0.6 ,H .5 0.0 0.0 a.u *_ 0.0 0.0 4_ (1.0 0.0 4 0.0 1 16 _ 30 19.tt 1.4 24 H VC 50 p(>ui 7.7? .47 1.22 * 23 4b. 0 4 1.2 lb.9 ,H 62.6 2.2 20. r * .3 33.1 .It 6.4 o> 5.2 * .4 0.5 9 0.3 * 2.1 2.6 0.0 * 0.0 .1) 4 .5 0.0 4. 0.0 .5 * 0.0 0.0 .1 .t 64 4_ It 22.0 * 3. 1 v*; 250 |I|>M 7.12 0 1 . lb .44 (31 .21 (3) 44.1 4 14.4 64.0 * 3.0 . 4.H (11 (3) (3) 21 .5 * 2.1 (3) 33.4 * . (3) 4.3 1 .5 (71 6.1 4_ 1 .5 <11 10.3 * 1.7 MS . II 2.0 0.0 4 0.0 .5 4_ .5 0.0 * U. 0 .t> 4 .s 0.0 0.0 0.0 * 0.0 1 ? 7 * 13 26.0 4 3.0 vc 1,000 ppu 6.24 t 1 .54 4 42.7 4 14.0 * 69.2 .77 (3) . *0 (31 3.S (31 1.1 (31 3.5 (31 22.7 1.4 (31 32.0 4 6.0 .5 (31 .5 (21 4.2 .7 (31 H.B 4_ 3.3 91.0 * 3.5 0.0 4 0.0 0.0 *_ 0.0 0.0 4 0.0 .3 4 .3 0.0 4_ 0.0 . 0.0 4 0.0 66 4 6 20. H 166 1.0 MU Entries are mean + S.E. of 4 mice except as noted in parentheses. vuc |<|ua 5.H J * 1 .411 * .64 (3) .25 (3) 34.3 12.4 67.7 * 3.2 1.0 1 .6 (31 (3) (3) 21.3 31 .5 4 . 1 (31 .6 (3) 3.1 * .I 12. J * Mb. 3 * 5. 1 4.4 0 .0 * 0.0 * .6 0.0 4 (1.0 1.0 .4 0.0 * 0.0 U.O *_ 0.0 1 05 * 1 6 29.ll 4 2.4 06 t 60 (31 SL 083895 TABLE 47 LABORATORY DATA OF MALE MICE EXPOSED TO VC OR VDC FOR 6 MONTHS OOStJ PPH 63 f Hi IHKOCY IF.S 1X10 /HH t HE 1ICULOCYTES. k CONTROL 7.21 ft .31 .97 ft . 2A 50 PPM VC 7.20 .79 ft .22 .17 ISO PPH VC f.39 ft .12 1. 10 ft .18 HEHA10CHIT. VOL. * HEMOALOHIN. OH. k HCV. CUBIC MICHONS HCHU, HICHO MlCHOOHS. AS.5 l.A 1A .2 .S 03.2 19. S ft 1.2 .A AA ,S 13.0 ft 61.3 ft IB.8 ft 1.2 .6 .3 .2 A 7.3 1 A # A ft 6A.0 ft 19.S ft .9 .3 .1 .A MCHHCt OH k 53 PLATELETS 1X10 /HH 1 33 LEUKOCYTES 1X10 /HH t 31.3 ft .7 3.1 ft .7 8.A ft 3.9 30.6 6.A .5 .9*' S.2 1 .1 30.5 6.2 4 6.0 ft .7 .A l.A NEUTHOPHILS. k 20.8 ft A.6 26.5 * A.O 37.0 10.1 LYMPHOCYTES. k 79.0 _ A.7 7 1.5 A .0 6P.8 ft 10.5 BANDS* k 0.0 0.0 0.0 * 0.0 0.0 0.0 EOSINOPHILS* k 1.0 .A 0.0 ft 0.0 .3 ft .3 BASOPHILS, k 0.0 ft 0.0 0.0 0.0 0.0 0.0 MONOCYTES* k 0.0 * 0.0 0.0 *_ 0.0 0.0 ft 0.0 AlYPJCALt k NUCLEATED HliC* k 0.0 ft 0.0 0.0 ft 0.0 0.0 1 0.0 0.0 ft 0.0 0.0 0.0 0.0 ft. 0.0 SOP T * IU/L SB.I ft 11.) (31 A9.3 ft 16.9 131 6A.H ft 10.5 BUN. HO k hachophaoes A9 INO./HM 1 28.3 ft A.A 77 ft 39 131 28.0 * 7.0 131 17.0 ft 2.8 ENTRIES ARE HEAD A STANDARD ERROR OF 4 HICK EXCEPT AS NOTED IN PARENTHESES. a/ Significantly different from Control group, p < 0.05, Dunnett'a eultlpla comparison procedure. 1,000 PPH VC 6.25 .26 1.10 39.3 ft 11.7 . 13 2.3 a/ .6 a/ 62.7 1.3 18.8 ft .A 30.0 ft .6 5.1 ft .7 A.l 51.5 A 7.8 ft .7 5.8 a/ 5.9 a/ 0.0 0.0 .8 ft. .5 0.0 ft. 0.0 0.0 ft 0.0 0.0 0.0 0.0 ft 0.0 83.5 ft 18.6 38.5 17.6 6?9 193 55 m vuc 6.76 4_ .35 1.69 .35 A 2.5 13.1 1.3 .5 63. 1 2.1 19.5 .5 30.9 *. .5 A.5 * .8 3.A 35.5 .7 A.A 6A.5 O.A 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 * 0.0 0.0 4 0.0 96.5 * 17.8 AO.8 9.9 353 173 ( SL 083896 TABUi 48 LABORATORY DATA OF FEMALE MICE EXPOSED TO VC OR VDC FOR 6 MONTHS post I PPM 63 EHY ItIHOCYTl S I M 10 /MM ) RETICULOCYTES, % HEHATQCR If, VOL. % HEMOGLOBIN, GH. % HCV, CUBIC MICRONS HCHII' MICHO HICROGHS. MCtmCf CM % 53 PLATELETS IXl# /MM I 33 leukocytes ixie /mm i nuitrgphils. * 00 lymphocytes* % to HAMITS, % EOSINOPHILS. % IIAS0PH1LS* * MONOCYTES* * atypical. * NUCLEATE!) RltC. * SOPI IU/L HUN. MG % 3 MACROPHAGES INO./MM I TROL .37 121 .IV (3t 1.5 42) .5 4 2t 1.3 421 .2 421 .2 121 0.0 <21 .3 431 A . T 43) A.9 131 0.0 1.5 * .9 0.0 0.0 0.0 0.0 6 13) 1.7 13) AO <31 50 PPM VC 7.A? * .00 42) 1.3A * .10 A A .5 * A.5 12) tA .3 I.T 42) 59.0 .6 12) 19.3 t .5 42) 32.2 * 3.9 * .5 42) .9 42) 3.9 1 1.3 12) 25.3 4). A 7A.3 0.1 0.0 1 0.0 .5 i .5 0.0 * 0.0 0.0 * 0.0 0.0 1 0.0 0.0 1 0.0 51 i 25 <21 21. B t 2.1 250 VC 7.35 * .25 90 4 .19 A ft , 0 4 1.1 1 A ,9 4 .14 65.A 4 1 .0 20.3 #_ .5 31.0 A. 6 1.1 .A 7.A .7 20.0 A.A 79.0 4 5.0 0.0 4 0.0 1.0 *_ .7 0.0 * 0.0 0.0 0.0 0.0 0.0 0.0 0.0 17 6 42) 1A.0 1,0 <21 1,000 PPM VC 7.31 i .15 1 A1 i .30 A7.0 * 1,9 1A . 6 -i- .5 6A.5 t 3.5 20.1 i- 1.0 31.2 * .6 S.O * .ft 6.A * l.ft 31.5 * 67.a * 7.1 6.a 0.0 * 0.0 .ft i- .5 0.0 0.0 * 0.0 0.0 0.0 * 0.0 i 0.0 0.0 51 * 5 ia.3 1 A 00 * 1.3 AS IJI^ 55 PPM VDC 6.26 1.02 1 .A5 _ . A9 39.3 * 6.ft 12.A 62.2 4 2.0 1 .9 19.ft 31 .11 4 .3 .6 A .6 * A.0 .3 .7 23.ft 3. 1 76.0 * 0.0 3.2 0.0 .3 0.0 .3 0.0 0.0 * 0.0 0.0 * a.a * 11)7 * 0.0 0.0 29 31.3 10.7 235 * H3 ENTRIES AllE MEAN 1 STANDARD ERROR OP A MICE EXCEPT AS NOTED IN PARENTHESES. a/ Significantly different froa Control group, p < 0. OS, IXinnett'a aultlpla cmparlaun procadure. SL 083897 SL 083898 TABLE 49 LABORATORY DATA OF MALE MICE EXPOSED TO VC OR VDC FOR 9 MONTHS ooset ppm *i erythmocvies ixio tm i Control 6.1) t .92 HEI1CUL0CYIES. * HlMAlOCRtl. VOL. % HEMOGLOHIN. OH. % HCV. CtlHlC MICRONS l.*S t 1.41 39.5 t 4.0 12.1 i I.T 6S.S 2.4 HCHH. MICRO H1CR06MS. NCHHC. AH S] PLAIELETS (Ml* /MM 1 13 LEUKOCYTES INI* /MM 1 neutrophils. LYMPHOCYTES. % HANDS* % EOSINOPHILS. % BASOPHILS. 19.9 i 1.2 30.4 i 1.1 4.T 4_ 4.2 i 4*.5 .3 .0 0.1 59.0 i o.s M 0.0 s 4_ .3 MONOCYTES, t M* atypical. * 1.1 t *.# NUCLEATED MAC* l.t l 1.1 SGPI. IU/L MUM* Mfi % 30 i 20.S 4 4 S.2 MACROPHAGES INO./HH 1 no t 25 wimiBS ark wh i otahdau aim of row hick. JO PPM VC 5.TO * .21 |.HO 4 .19 41.3 1.0 13.V * T1 3 4_ 2.4 24.0 4 A 31.T 4 .2 4.1 1 5.0 4 .9 .6 33.5 4 3.6 65.3 4_ 4 T 4.0 4 0.0 1.1 4 0.0 1.1 4_ 0.0 0.0 4 0.0 o*o 4^ 0.0 0.0 1 0.0 IT 4 45 19.0 2.3 IS t 4 m 6.66 *_ VC .20 2.50 * .46 43.3 L 12.H 4 2.0 .6 66#e 4 3.2 19.6 4 29.0 4 1.3 2.1 4.1 t 4.4 4_ .6 .9 23.0 L T6.0 4 2.T 2.0 0.0 0.0 t 0.0 0.0 0.0 4 0.0 .1 4_ O.A 4 0.0 * .3 0.0 0.0 fid * 14 20.) * 44 t I.T j\f Significantly different from ilia cimtrol group, t < 0.05, ftunaatt'a uhi^li coaptrlioti procedure, 1000 DM VC 5.34 4 1.00 6.62 t 33.3 4 10.0 4 1.24 4.ft 1.5 65.5 4 6.2 21 .5 4_ 12.T 4 2.2 .6 4,1 4 .T 3.3 i 4?.ft 4 1.3 6.T 6T.0 4 6.5 0.0 1 0.0 0.0 4 0.0 0.0 1 0.0 .3 1 0.0 4 0.0 4 .3 0.0 0.0 4 23 60.3 4 23.3 031 4 162 *' 55 wc 6.S6 4 1? 1 .96 4 ft 42.5 4 .9 13.3 #_ 4 65.2 4_ 2.7 20.S 1.1 31.4 ft 5.5 4_ 6 4*6 4_ 1.6 39.0 4_ 4.1 60.5 i .3 4 1.9 .3 .3 * 0.0 4 3 0.0 0.0 4_ 0.0 4 0.0 * 0*0 0*0 0.0 154 49 *2.3 4 50 4 3.9 ft SL 083899 TABLE 50 LABORATORY DATA OF FEMALE MICE EXPOSED TO VC OR VDC FOR 9 MONTHS oosfi ppm 6% tWTlMWOCVIfS 1*1* /MM 1 on icwoctH s. * HEMATOCMIT. VOL. * HLHOGLOHlN. I.M. * HCV. CUBIC MICHUNS MCHH* MICHO MICPOr.MS. MCHHCt GM S3 PLAIELMS 1*1* /MM 1 1J LFUKOCVHS 1*10 /MM 1 nfuimuphilS' LYMPHOCYTES. * farms* * EOSINOPHII S. * BASOPHILS* MONOCYTES* * ATYPICAL* * NliCLLATFO HOC, S SGPI* 111/L HIM* wn % tacMtPtuf'ts iwo./hh Control 6.A4 * .ss 1.31 * . is **.3 11. M * 65.b * I." .S l.H 2R.S I.A 31.? A.? * 2.S * A 1.0 1 .1 *2.0 S.? SB .0 s.? 0.0 * rt.ft 0.0 0.0 0.0 0.0 0.0 0.0 0.0 *_ 0.0 0.0 0.0 S3 ?7 2A.S ?.b A3 j* A3 SO pp. VC A* os *_ -S3 I.A? t 44.0 n.s * .11 1.3 .4 AS 1 * A.ft ?*.l 30.0 * *. * ?.ft * 1 .S .? ? .6 3b.S A3.S *_ 3.0 3.0 0.0 * o.n 0.0 41.0 0.0 0.0 0.0 a.o 0.0 0.0 a.a 0*0 4? s ?3.S SO i.? lb 250 f>pw VC A. 3" 6.6 1 30.0 10.S 6rt.S ?4.fl 3S.0 2.5 9.S 30.0 0.0 1.0 0.0 0.0 0.0 0.0 ss |.U 245 I00U ppm VC S.AA 2.?3 1*. ft !?.b 31.3 23.2 3?. 3 2.3 A.3 3A.0 0.0 0.0 0.0 0.0 0.0 0.0 46 lft.0 ??s emvtES ARC MEAN L S1AHOARD ERROR Of FOUR HICK EEC EFT FOR THE ISO PP" AMO 1000 Fp. VC CROWS IN WHICH OHCV OHM HOUSE WAS THMINATED IN EACH CROUP. SS ppa vuc 7.0. 0. i . M.l 1. fit i .10 46. t .ft 14. H i .4 66. ft * 3. ft 21.A t ?.o 11 H 1.) . 4.1 t . i..' .1 .. 1 13.3 * 4.9 66. H * A.9 <1.0 1 0.0 o.o II .0 0.0 1 0.0 0.0 * 0.0 o.o i 0.0 0.0 * 1A1 I 0.0 6? 111. 1 4.4 16 i 1 0 i TABLE St LABOBATOBY DATA OK MALE MILE EXPOSED TO VC OB VDC FOB 12 HOHTIIS EHYTItHOCYTES (Kit /MM 1 7.39 .09 7.88 .19 7.50 .26 RETICULOCYTES. ft HEMATOCRIT. VOL. ft HEMOGLOMIN. CM. * HCV. CUIlIC MICRONS MCHU* MICRO MfCNOGHS. 3,05 1.53 53.0 .5 13.2 t 58.2 .1 .G 17.9 .2 2.56 *_ .35 55.0 1.2 13.9 55.9 17.7 .6 1.0 .1 2.5b 50.6 12.8 .30 171 3.7 171 1.1 55.3 * 5.7 (71 17.3 * 1.5 MCHHCt CM ft 53 PLATELETS 1X10 /MM > 33 LEUKOCYTES <XI0 /MM 1 NEUTROPHILS. ft LYMPHOCYTES. ft RAMOS. * 30.8 L .2 7.9 .7 5.8 27.8 1.0 5.6 72.3 0.0 5.6 0.0 31.7 6.1 .9 .3 8.9 1.7 36.0 7.6 63.7 * 7.5 0.0 0.0 ` 30.0 .6 171 7.5 5.3 .7 (71 .8 28.8 71.3 5.0 5.0 0.0 0.0 EOSINOPHILS, ft 0.0 0.0 0.0 0.0 0.0 0.0 BASOPHILS, ft 0.0 0.0 0.0 0.0 0.0 0.0 MONOCYTES, ft 0.0 1 0.0 .3 t .3 0.0 0.0 ATYPICAL, ft 0.0 t 0.0 0.0 0.0 0.0 0.0 NUCLEATED BBC, ft co S6PT. IU/L DUN. MG ft 0.0 t 0.0 51.8 1 6.9 25.8 *_ 3.9 52.3 1 5.9 30.0 2.6 0.0 * 0.0 _ / 99.0 t 16.2 171, a/ 51.5 t 5.6 171- machophages ino./mm > 25 6 395 325 (2) 135 53 083900 Entries are aiean + standard error of 5(Control), 1(50 ppw VC) or 8(55 ppa VOC) nice unless otherwise Indicated In parentheses. ml Significantly different Iron the control group, P <0.05, Dunnett's swltlple cnaparlson procedure. J TABLE 52 LAIlOflATORY BATA OP KtHAIiS Ml fit EXPOSED TO VDC FOtt 1 2 MONTHS ERYTHROCYTES 1110 /MM 1 RETICULOCYTES. * 7.72 1 .SO * .15 .20 7.39 2.20 .30 a/ .25 HEMATOCRIT* VOL. * HEMOGLOBIN, GM. ft HCV. CUBIC MICRONS HCItU, MICRO MICHOGMS. HCliHC. GM ft 53 PLATELETS 1X10 /MM 1 33 LEUKOCYTES 1X10 /MM I NEUTROPHILS* * LYMPHOCYTES* * QaOt BANDS* ft EOSINOPHILS* ft BASOPHILS* ft HOMOCYTES. ft ATYPICAL, ft 43.9 13.9 .7 .3 56.9 .9 1A.0 .2 31.6 1 .4 6.2 .6 4.9 20.7 .0 4.2 70.6 4.1 0.0 0.0 .3 .2 0.0 0.0 .4 t .2 0.0 0.0 43.1 1.6 13.6 .6 SB. 4 .4 IB.4 *_ 31.S .2 .4 7.1 t 3.6 1.4 .11 32.5 4.0 67.3 *_ 4. 1 0.0 0.0 .3 .2 0.0 0.0 0.0 0.0 0.0 0.0 NUCLEATED RBCt ft SGPT. 1U/L BUN, HG ft 3 MACROPHAGES INO./HH 1 0.0 0.0 103 23.1 16 1.5 SB *_ 16 0.0 0.0 7ft 5 36.7 4.6 22 L 5 -/ Entries are Kean + standard error of 10 ntce(Coatrol) or ft nlce{55 ppM VDC) unless otherwise Indicated In parentheses. / Significantly different from the control group, t <0.05, Dunuett'a Multiple coMpsiison procedure. SL 083901 TABLE 53 NUMBERICAL DISTRIBUTION OF CHROMOSOMES IN BONE MARROW OF MICE EXPOSED TO VC OR VDC Treatment No. of Mice Chromosome Freauencv 38 39 40 41 a 42 Tetraploids (4N) per 100 cells After One Month Control 6 5*/ 5 36 3 1 0.42 0.20^ VC (1,000 ppm) 6 6 4 39 1 0 0.25 0.17 VDC (55 ppm) 6 2 5 40 2 1 0.25 0.17 After Three Months Control 4 5 7 37 1 0 0.0 0.0 VC (1,000 ppm) 4 6 6 37 1 0 0.0 0.0 VDC (55 00m) 4 6 6 35 3 0 0.15 0.15 After Six Months Control 6 7 4 37 1 1 0.50 0.22 VC (1,000 ppm) 6 4 4 34 6 2 0.33 0.21 VDC (55 ppm) 5 5 7 36 2 0 0.36 0.22 After Nine Months VC (1,000 ppm) 5 2 3 42 2 1 0.0 0.0 After Twelve Months VDC (55 ppm) 6 2 5 41 1 1 0.17 0.17 j/ Mean b/ Mean S.E. SL 083902 87 TABLE 54 MORPHOLOGICAL ABERRATIONS OF CHROMOSOMES IH BONE MARROW OP MICE EXPOSED TO VC OR VDC Tree trnen t No, of Chromatid Chromatid Translocations Total Aberrations Mice Gaos per 50 Celia Breaks ner 50 Celia per 50 Cells per 50 Cells After One Month Control VC (1,000 ppm) VDC (55 ppm) 6 6 6 0.67 0.3^ 0.52 0.23 0.97 0.26 0.0 0.0 0.17 0.17 0.33 0.21 0.20 0.20 0.17 * 0.17 0.17 * 0.17 0.86 0.31 0.86 0.13 0.47 0.43 After Three Months Control VC (1,000 ppm) VDC (55 ppm) 4 4 4 0.25 0.25 0.0 0.0 0.50 0.50 0.0 0.0 0.25 0.25 0.25 0.25 0.0 * 0.0 0.0 ^ 0.0 0.0 0.0 0.25 0.25 0.25 0.25 0.75 0.48 After Six Months Control VC ( 1,000 ppm) VDC (55 ppm) 6 6 5 0,0 0.0 0.0 db 0.0 0.0 0.0 . 0.0 0.0 0,0 0.0 0.0 0.0 0.17 0.17 0.17 0.17 0.0 * 0.0 0.17 0.17 0.17 0.17 0.0 0.0 (continued to next page) SL 083903 Treatmen t TABLE 54 (Concluded) Ho. of Chromatid Chromatid Translocations Total Aberrations Mice Caps per 5Q Cells Breaks per 50 Cells per 50 Cells per 50 Cells After Nine Months VC (1,000 ppm) 5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 After Twelve Months VDC (55 ppm) 6 0.33 0.21 0.0 0.0 0.0 0.0 0.33 0.21 GO vO ............................................ ............. .......... a/ Mean S.E. SL 083904 TABLE 55 ORGAN WEIGHTS OF MALE MICE AFTER EXPOSURE TO VC OR ypc FOR 1 MONTH Control VC 50 ppm VC 250 ppm VC 1.000 ppm VDC 55 ppm Absolute Weight {gm! Terminal Body Weight Brain 33.0 1.0^ 0.51 0.02 31.0 1.2 0.47 0.01^ 32.0 0.7 32.5 0.6 29.8 0.8 0.47 0.01^/ 0.46 0.01-^ 0.46 0.00^ Liver 1.77 0.24 1.34 0.13 1.35 0.12 1.31 0.02 1.35 0.03 Kidney Spleen 0.54 0.03 0.17 0.02 0.51 0.04 0.13 0.02 0.51 0.02 0.14 0.01 0.54 0.0L 0.13 0.01 0.46 0.02 0.11 0.01-^ lO Tes tea o 0.26 0.04 0.23 0.01 0.24 0.01 0.22 0.02 0.24 0.01 Relative to Brain Weiftht Cgm/gm) Liver Kidney Spleen Teatea 3.5 0.5 1.06 0.08 0.34 0.05 0.52 0.08 2.9 0.3 1.09 0.08 0.27 0.04 0.49 0.03 2.9 0.3 1.11 0.06 0.30 0.02 0.52 0.02 2.8 0.0 1.17 0.04 0.27 0.02 0.49 0.04 2.9 0.1 1.01 0.05 0.24 0.02 0.52 0.03 a/ Mean S.E. of four mice. b/ Significantly different from control, p <0.05, Dunnett'a Multiple Comparison procedure. Cfi X? 083905 ll TABLE 56 ORGAN WEIGHTS OF FEMALE MICE AFTER EXPOSURE TO VC OR UPC FOR l MONTH Terminal Body Height Brain Liver Kidney Spleen Ovaries Liver Kidney Spleen Ovarlea Control 23.3 l.l5/ 0.48 0.01 1.13 0.08 0.33 0.03 0.10 0.01 0.02 0.01 2.36 0.18 0.69 0.05 0.21 0.02 0.05 0.16 VC 50 ppm VC 250 ppm VC 1.000 ppm Absolute Weight (rip) 27.8 * l.A 0.47 0.01 1.15 0.07 0.38 0.02 0.15 0.02 0.01 0.00 26.5 1.5 0.47 0.01 1.12 0.02 0.37 0.02 0.13 0.02 0.02 0.00 24.8 0.9 0.49 0.01 1.03 0.06 0.36 i 0.01 0.14 0.03 0.02 0.00 Relative to Brain Height (rdi/rhO 2.43 0.12 0.81 0.03 0.31 0.04 0.03 0.01 2.40 0.06 0.79 0.04 0.27 0.03 0.03 0.01 2.13 0.07 0.74 0.01 0.30 0.08 0.05 0.01 VDC 55 ppm 26.5 1 0.9 0.49 0.01 1.31 0.06 0.39 0.01 0.13 0.01 0.02 0.00 2.69 0.08 0.80 0.00 0.27 0.02 0.03 0.01 cn a/ Mean S.E. of four mice. b/ Significantly different from control, p < 0.05, Dunnett's Multiple Comparison procedure. 083906 TABLE 57 ORGAN WEIGHTS OF MALE MICE AFTER EXPOSURE TO VC OR VDC FOR 2 MONTHS Control VC 50 ppm VC 250 ppm VC I.000 ppm VDC 55 ppm Absolute WeiRht (gm) Te rtnina 1 Body Weight 33.5 O.W 33.8 1.9 34.3 1.7 36.8 1.4 32.0 0.6 Brain 0.50 0.01 0.47 0.04 0.51 0.01 0.48 0.03 0.49 0.02 Liver 1.72 0.13 1.50 0.08 1.50 0.07 1.57 0.09 1.60 0.04 Kidney 0.69 0.04 0.70 0.08 0.66 0.03 0.69 0.02 0.52 0.04 VNOJ Spleen Testes 0.11 0.04 0.27 0.02 0.15 0.03 0.26 0.01 0.15 0.01 0.28 0.02 0.16 0.00 0.26 0.02 0.11 0.02 0.26 0.02 Relative to Brain Weight (gm/gni) Liver Kidney Spleen Testes 3.A 0.3 1.37 0.08 0.23 0.06 0.5A 0.03 3.3 0.4 1.56 0.29 0.33 0.05 0.58 0.06 2.9 0.1 1.29 0.05 0.30 0.02 0.55 0.05 3.3 0.2 1.44 0.10 0.34 0.03 0.55 0.07 3.3 0.1 1.06 db 0.04 0.22 0.03 0.53 0.02 a/ Mean S.E. of four mice SL 083907 l TABLE 58 ORGAN WEIGHTS OF FEMALE MICE AFTER EXPOSURE TO VC OR VPC FOR 2 MONTHS Terminal Body Weight Brain Liver Kidney Spleen Ovaries Control 28.8 0.6^ 0.53 0.02 1.28 0.10 0.43 0.02 0.14 0.01 0.03 0.00 VC 50 ppm VC 250 ppm Absolute Weight (gup 29.0 1.9 0.53 0.02 1.29 0.07 0.42 0.01 0.13 0.01 0.04 0.01 29.3 0.6 0.47 0.01 1.25 0.04 0.39 0.01 0.17 0.01 0.03 0.00 VC 1,000 ppm 29.5 1.2 0.54 0.02 1.28 -t 0.07 0.44 0.02 0.14 0.01 0.03 0.00 VDC 55 ppm 28.0 0.4 0.51 0.01 1.32 0.08 0.42 0.01 0.12 0.01 0.04 0.01 UVO) Liver Kidney Spleen Ovaries 2.44 0.16 0.81 0.03 0.26 0.01 0.06 0.01 Relative to Brain Weight fgm/gnO 2.44 0.09 0.79 0.03 0.24 0.02 0.07 0.01 2.66 0.11 0.84 0,03 0.36 0.03-' 0.07 0.00 2.40 0,12 0.82 + 0.05 0.26 0.02 0.06 t 0.00 2.60 0.17 0.82 0.01 0.23 0.02 0.07 0.00 a/ Mean + S,E. of four mice. b/ Significantly different from Control, p< 0.05, Dunnett*s Multiple Comparison procedure. SL 083908 TABLE 59 ORGAN WEIGHT'S OF MALE MICE AFTER EXPOSURE TO VC OR TOC FOR 3 MONTHS Control VC 50 ppm VC 250 ppm VC 1.000 ppm VDC 55 ppm Absolute Weight (gnQ Terminal Body Uelght 34.9 el*' 34 .8 1 1.,1 32.8 1 .2 34.4 1.2 28.3 1.8*' Brain 0.52 0.02 0. 49 iL 0,,01 0.51 0 .02 0.52 0.01 0.51 i 0.01 Liver 1.64 0.09 1. 54 0..08 1.46 0 ,06 1.62 0.08 1.50 0.07 Kidney Spleen Ml -P> Testes 0.63 0.03 0.12 0.01 0.22 0.02 0. 67 0,,06 0. 16 0,,02 0. 30 0..03 0.64 0 .05 0.14 0 .01 0.28 0 .01 0.68 0.03 0.12 0.01 0.27 0.02 0.50 0.01 0.12 i: 0.01 0.27 0.02 Liver Kidney Spleen Tes tes 3.19 0.14 1.23 0.04 0.23 0.01 0.43 0.03 Relative To Brain Weiftht (w/w) 3.16 0.13 1.37 0.10 0.33 0.04^ 0.62 0.06*' 2.88 0.19 1.25 0.07 0.27 0.02 0.54 * 0.02 3.13 0.16 1.31 0.07 0.24 0.01 0.52 0.04 2.97 0.18 0.99 0.04 0.23 0.02 0.55 0.05 af Mean S.E. of four mice. b/ Significantly different from control, p < 0.05, Dennett's Multiple Comparison procedure SL 083909 I J1 1J TABLE 60 ORGAN WEIGHTS OF FEMALE MICE AFTER EXPOSURE TO VC OR VDC FOR 3 MONTHS Terminal Body Weight Bra in Liver Kidney Spleen Ovaries Control 25.9 t 1.5^ 0.49 0.01 1.18 0.10 0.50 0.14 0.12 0.01 0.04 0.01 VC 50 ppm VC 250 ppiu Absolute Weight fain't 29.4 1.2 0.51 0.02 1.27 0.06 0.41 0.02 0.11 0.01 0.04 0.01 26.0 1.1 0.55 0.01 1.12 0.07 0.38 0.04 0.14 0.02 0.04 0.01 VC 1.000 _RTM 26.9 0.7 0.51 0.03 1.21 0.07 0.45 0.07 0.14 0.01 0.04 0.01 VDC 55 _R5 25.9 0. 8 0.47 0. 03 1.32 0. 07 0.41 0. 01 0.13 0. 00 0.04 0. 01 Relative to Brain WeiRht ifftm/Rm) Liver Kidney Spleen Ovaries 2.38 0.17 1.01 0.27 0.24 0.01 0.07 0.01 2.49 0.04 0.81 0.03 0.22 0.01 0.09 0.01 2.03 0.10 0.70 -t 0.05 0.24 0.03 0.08 0.01 2.40 0.21 0.89 0.11 0.27 0.02 0.08 0.03 2.81 0. 11 .0.87 0 05 0.28 0. 02 .0.10 0 02 a/ Mean S.E. of four mice. TABLE 61 ORGAN WEIGHTS OF MALE MICE EXPOSED TO VC OR VDC FOR 6 MONTHS Control 50 ppm VC 250 ppm VC 1,000 ppm VC 55 ppm VDC Absolute Weight (riw) Terminal Body Weight Brain Liver Kidney Spleen Testes 38.2 + l.ja/ 0.50 + 0.01 1.89 + 0.05 0.63 + 0.06 0.14 + 0.01 0.29 + 0.01 36.1 + 3.3 0.52 + 0.01 1.73 + 0.17 0.68 + 0.03 0.16 + 0.04 0.28 + 0.04 38.5 + 0.9 0.52 + 0.04 1.81 + 0.08 0.74 + 0.04 0.15 0.02 0.29 + 0.02 36.7 + 0.5 0.55 0.02 1.75 + 0.14 0.61 0.04 0.22 + 0.04 0.33 + 0.09 Relative to Brain Weight (gm/gm) Liver Kidney Spleen Testes 3.8 + 0.2 1.27 + 0.12 0.29 + 0.02 0.58 + 0.03 3.3 0.4 1.32 + 0.07 0.32 + 0.08 0.54 + 0.07 3.5 + 0.3 1.45 + 0.13 0.31 + 0.06 0.56 + 0.03 3.2 + 0.2 . 1.11 + 0.09 0.39 + 0.06 0.61 + 0.17 36.5 1-4 0.50 + 0.01 1.94 + 0.05 0.57 + 0.02 0.16 + 0.02 0.27 + 0.00 3.9 + 0.1 1.15 + 0.05 0.32 + 0.04 0.54 + 0.02 a/ Mean + S.E. of four mice. SL 083911 TABLE 62 ORCAH WEIGHTS OF FEMALE MICE EXPOSED TO VC OR VDC FOR 6 MONTHS Group Control 50 ppm VC Absolute Weight 250 ppm VC (rpi)__________________________________________________________ 1,000 ppm VC 55 ppm VDC Terminal Body Weight Brain Liver Kidney Spleen Ovaries 33.7 + 1.9/ 0.51 + 0.02 1.63 + 0.10 0.45 + 0.04 0.17 + 0.02 0.05 + 0.01 31.0 + 1.3 0.51 + 0.03 1.43 + 0.07 0.42 + 0.02 0.19 + 0.03 0.03 + 0.00 30.4 + 1.8 0.52 + 0.01 1.60 + 0.10 0.50 + 0.01 0.19 + 0.02 0.06 + 0.01 31.9 + 1.3 0.52 + 0.02 1.53 + 0.17 0.46 + 0.04 0.18 + 0.02 0.04 0.01 vo Liver Kidney Spleen Ovaries Relative to Brain Weight (gut/gin) 3.2 + 0.3 0.90 + 0.08 0.34 + 0.04 0.09 + 0.02 2.8 + 0.3 0.84 + 0.08 0.37 + 0.05 0.07 + 0.01 3.1 0.2 0.98 0.04 0.37 0.03 0.11 0.02 2.9 + 0.2 0.88 + 0.05 0.35 + 0.05 0.09 + 0.01 31.7 + 1.4 0.53 + 0.02 1.56 + 0.14 0.49 + 0.02 0.19 + o'oi 0.04 + 0.00 3.0 + 0.3 0.95 + 0.06 0.36 + 0.03 0.08 + 0.01 a/ Mean + S.E. of four rats. SL 083912 TABLE 63 ORGAN WEIGHTS OF MALE MICE EXPOSED TO VC OR VPC FOR 9 MONTHS Control 50 PPM VC 250 PPM VC 1.000 PPM VC 55 PPM VPC Absolute Weight (gm) Terminal Body Weight Brain Liver Kidney Spleen Testes 39 + 3a/ 0.54 + 0.02 1.74 0.12 0.75 + 0.17 0.17 + 0.04 0.36 Jm 0.03 37 2 . 0.48 0.02- 1.49 + 0.12 0.70 + 0.05 0.16 + 0.02 0.23 + O.OlM/ 38 + 1 0.50 X 0.01 1.59 X 0.09 0.69 0.03 0.20 + 0.02 0.28 + 0.02 37 + 3 38 + 1 0.53 + 0.01 0.47 1.71 0.15 1.93 + 0.66 + 0.03 0.57 + 0.0^ 0.31 0.09 0.19 + 0.03 0.27 i 0.03--^ 0.30 + 0.02 Relative to Brain Weight (g/a) Liver Kidney Spleen Testes 3.2 + 0.2 1.38 + 0.26 0.31 0.06 0.68 + 0.08 3.1 + 0.3 1.47 + 0.11 0.34 + 0.02 0.49 + 0.03 3.2 + 0.2 1.39 + 0.07 0.39 + 0.04 0.56 + 0.03 3.3 0.3 1,26 + 0.06 0.58 + 0.17 0.51 + 0.06 4.1 j. 0.4 1.20 + 0.07 0.40 + 0.07 0.64 4* 0.05 a/ Mean + S.E. of four mice. b/ Significantly different from Control group, p < 0.05; Dunnett's multiple comparison procedure. 98 SL 083913 TABLE 64 ORGAN WEIGHTS OF FEMALE MICE EXPOSED TO VC OR VPC FOR 9 MONTHS Control 50 PPM VC 250 PPM VC 1.000 PPM VC 55 PPM VPC Absolute Weight (gm) Terminal Body Weight Brain Liver Kidney Spleen Ovaries 34 + 20.54 i 0.01 1.36 + 0.09 0.47 + 0.01 0.14 + 0.01 0.07 + 0.01 32 t 3 0.54 + 0.02 1.31 + 0.13 0.49 + 0.04 0.15 0.02 0.05 0.02 35(1) 0.43(1) 1.93(1) 0.45(1) 0.55(1) 0.05(1) 30(1) 0.51(1) 1.20(1) 0.37(1) 0.11(1) 0.02(1) Relative to Brain Weight (gm/gm) 30 + 1 0.50 0.03 1.34 + 0.05 0.46 + 0.03 0.16 t 0.01 0.05 + 0.01 Liver Kidney Spleen Ovaries 2.54 + 0.16 0.38 + 0.04 0.25 + 0.01 0.14 + 0.02 2.41 t 0.15 0.90 4 0.05 0.27 + 0.04 0.10 0.03 4.04(1) 0.93(1) 1.14(1) 0.10(1) 2.35(1) 0.72(1) 0.22(1) 0.05(1) 2.68 + 0.08 0.92 t -04 0.33 + 0.02 0.10 + 0.01 a/ Mean + S.E. of four mice except as noted in parentheses. 99 St> TABLE 65 ORGAN WEIGHTS OF MALE MICE EXPOSED TO VC OR VDC FOR 12 MONTHS Group Brain Liver Kidney Spleen Testes Absolute Weight (ga) Control 50 ppm VC 0.490 + 0.019-/ (4) 2.1 + 0.0(4) 0.78 0.03(4) 0.17 + 0.02(4) 0.302 + 0.009(4) 0.493 + 0.020(3) 1.7 + 0.0(3) 0.73 + 0.03(3) 0.12 + 0.01(3) 0.283 + 0.020(3) 55 ppm VDC 0.479 + 0.010(8) 2.2 0-2(8) 0.57 + 0.02-- (8) 0.26 + 0.09(8) 0.269 + 0.016(8) Relative to Brain Weight (gm/gm) Liver Kidney Spleen Testes 4.3 + 0.2(4) 1.59 + 0.02(4) 0.36 + 0.04(4) 0.62 + 0.05(4) 3.6 0.1(3) 1.49 + 0.07(3) 0.25 + 0.02(3) 0.57 + 0.03(3) 4.6 0.4(8) 1.18 + o.o^^S) 0.53 + 0.18(8) 0.56 + 0.02(8) a/ Mean + S.E, of number of rats indicated in parentheses, b/ Significantly different from Control group, p <0.05; Dunnett's multiple comparison procedure. 100 SL 083915 TABLE 66 ORGAN WEIGHTS OR FEMALE MICE EXPOSED TO VC OR VDC FOR 12 MONTHS Group Brain Liver Kidney Spleen Ovary Absolute Weight (gm) Control 55 pom VDC 0.496 + 0.008^/ 1.38 + 0.06 0.44 + 0.02 0.121 + 0.007 0.04 + 0.01 0.466 + 0.015 1.63 + O.loV 0.46 + 0.02 0.150 + 0.015 0.06 + 0.02 Liver Kidney Spleen Ovary Relative to Brain Weight (gm/gm) 2.8 + 0.1 0.89 + 0.04 0.24 + 0.02 0.08 + 0.01 3.6 + 0.3-^ 0.98 + 0.05 0.32 + 0.03--7 0.14 + 0.05 / Mean + S.E. of ten rats (Control) or eight rats (55 ppm VDC). / Significantly different from Control group, p <0.05; Dunnett's multiple comparison procedure. laris* SL 083916 101 TABLE 67 SUMMARY OF BRONCHIOLO ALVEOLAR ADENOMAS IN HICE AFTER EXPOSURE TO VC OR Vjjg. Exposure i xtlig _ _ 1 month 2 months 3 months Control rf $ 0^ 0 00 00 SO ppm VC <S ? 00 00 10 250 ppm VC ? 00 10 10 1,000 ppm VC d" 00 12 2l 55 ppm VDC <t 00 00 00 a/ Humber of adenomas In four mice of each sex. SL 083917 ii i 1AIU.E 6# SUIHARY OF niums THAT OI.'OUHHF.D IKIK1NC 4 in 4 HJNITIS OF FXfoaOKF. m ou> it Include* only nice uhtck were found to have tuwn during A to 6 nonilia of txpomre to VC or VHC. y Severity of leelon: 1 - nlld; 2 - anderate; 1 - narked; A - severe; F - preii>nt; i questionable. c/ Pled or unechedulfld gemination. tn c* CP x e te o TABLE 691 SUMMARY OF TUMORS THAT OCCURRED IN MICE DURING 7 TO 9 MONTHS OF EXPOSURE TO 50 PPM VC.a/ Sex Mouse No. Tumor^ Week Terminated ______Male 92 10827 39 27 10539 ------------------------------c-f-----------cf 125 126 142" 141- 39 39 28 28 Female ________ 140^' 139- 138--^ 33 33 33 137S^ 136-^ 135s? 134Syf 34 35 36 36 Lung Bronchiolo alveolar adenoma Metastatic squamous 1 1 1i 1 1 1 1 ___ c e H _ca rc i noma______________ Liver _ Hemangiosarcoma _____________ Mammary Gland Duct adenocarcinoma Squamous cell carcinoma Anaplastic carcinoma 111 l11 13 23 l 23 1 23 _ Hemangiosarcoma _____________ 1 Epididymus and Teatea _ Hemangiosarcoma _____________ 1 Mediastinum Hemangioma______________________ 2 a/ includes only mice which were found to have tumors during 7 to 9 months exposure to 50 ppm VC. Five scheduled mice and two unscheduled mice were not found to have tumors, b/ Severity of lesion: l - mile; 2 - moderate; 3 - marked; 4 - severe; + - questionable. cf Died or unscheduled termination. [l iani: /u SUHHAWT OP TWOttS <<VPHE IH MCE fUMIHCi 7 JO 9 H^lllS (if KKlrjjHWtfc TQ 2M> IW T<u*oi~^ Sen Mouse Ho. Week Teralnatd ___ __________ tUile_______________ Itl 141 164 J#o/ 11& 11t& in-1 IH*-1 19 19 19 27 12 12 11 14 197! 19 _____ ____ __ _ -- ti'*2Je____ ____ ____ _____ _ . 21 \f 2\2'=j 21|SfJ Tjoil^ 2U9^ 2rtHlZ 2i>?/ 20fc^ 201? 204-^ 2IH-^ 202^ 201 ? 200' 27 27 lt> 1(1 II 12 12 12 12 11 11 14 14 14 Lung bi ottcliiolo-a Iveolur adenoma 3 2 2 HclMtNtlC I^UAHUUK call __ _ itC^noM ____ _ ______________ _ _ Liver _ U^f>UKi^aiciw|_______________ 2____________________1_______2 Mmury Cland |)uc( adenocercionais S4|uimiiib celt carclnoafl -- AnapIaa-t2c_c2r*iS2S*-- _ _ __________ _______________ _ _ * liniuL __________ _____ ________ Heart IlcwanKlotarcoaM ________________________________S Pancreas _ JJcaa|it||oaarcjHM _ ___________________________ Skeletal Muscle Ou _ Jojsrcoa*______ _ ________________ CastroIntestInal Tract _ ll^QKW^L^i________________ __________ 21 23_ ii 33 32 I2 i 2 A_______3_________A_______1________2_______l________3_______3________ 3_______2_________1_______3 33 13 2 3_ 3 I a/ Includes only slct which wars found to have tuasir* during 1 to 9 Months exposure to 2%0 pps VC. litas scheduled and two unscheduled silce were not found to have tusori. k/ Severity of lesion: 1 - mild; 2 - asMfterete; 3 * marked; A - severe; fr - quest(unable. c/ Hied or unscheduled termination. d/ (Inly female that survived 39 weeks exposure to 230 ppm VC. SL 083920 TABLE 71 SUMMARY OF TUMORS THAT OCCURRED TN MICE DURING 7 TO 9 MONTHS OF EXPOSURE TO 1.000 PPM VCA-- / Tumor--^ Sex Mouse No. Week Terminated Male 233 234 235 236 250^ 249--^ 247' 24 39 39 39 39 28 29 31 34 245--^ 244--^ 243--^ 24 2/ 34 34 34 35 24l/ 240/ 36 38 Lung Bronchiolo-alveolar adenoma 3 3 3 3 3 3 3 2 2 3 3 1J Liver Hemangiosa ream 4l2 31 332 3 33 o Malignant Lymphoma (continued to next page) SL 083921 It I Tuaur-^ Sea Houe No. Week Tanalnatad 269B&297& 27 TAflLC 71 (CimcUuli'il) a/SIIHHAHY OF THHUHS THAT OCCUKHLtt IH HICK DHKIHC I TO MOW IIS OF EXPOSOBE TO 1,000 PPM VC-' ______ ______________ 26t&t 211? 27 27 2d ftwle_______ ______________________________ ______ 2B2^ 261$/ 2J9/ 21&J 2 lid 2d 2* 30 II* 30 31 ll&J 31 34 274* 2711* 34 2 34 27\^ 270^ 2691^ 269A/ 34 36 36 37 Lung Bronchioloalveolar ad*......... 13233313 233)333)331 Hetaatatlc aquaaenia call care hum* 222 13 Metwatatlc anaplaatlc _ cdrciitoau __________ ___________________________________________________________________ __________________________________________________ _ ______3_ ________________________________________________ _ _ Liver ||ifMaiy;LoearciM __ _ _____J______|_______l__________________________1 _l_ _ _ ______)_ 2^______________________3______2__________________________________1__ _ ____1 _ H^awrir {iUnd Duet adenocarcinoma 1 I3 3 9<|uaai>ua call carcinoma II 3 o Auaplaatlc carcinoma -4 ilcwn^^oaarcoMa _ _ __________ __________________________)_____________ 33)22 3)22 _ _3_ }_____2______2_______ _ __ ________________ _ _ _ _____ _______ ___________ _____________________ ___________________________ _)__________________3____________ _____________3______________ ____________ ______________________________ Kidney Ikw-m^hiMrcoai ____ __ _______________________________________________________________i____________________________________________________________________________________ Heaentarlc Lymph Nodaa _ JlcMnngjojMrcoMa______________________________________________________________________________________________________________ 2________________________________ _____________________________________________ a/ Include# only mica which war# found to have twaora during } lu ) aumtUa eapoaurc lo l#WHi ppa VC. One unachedulcd termination waa nut iiHiiul to have any tiuaora. h/ Severity of lea Ion; 1 - ail Id; 2 - Moderate; 3 - awirked; 4 - ecverc; * - queertonabla. c/ Died ur unacliadulad termination. d/ Only female eurvlvlng 39 weak# aipuaura to 1,000 jipa VC. SL 083922 TABJ.K II SIHUH1 Of IUHOK3 TBI IJCClHlPro IK HICE BUMHjL Id 111 U HHHniS EMfUSIJIlE U> jfC yg 1>L-J Houma Ho. _______________________________ jg rr? Hole 91 9b 91 104^ lOI&f 11 11 11 41 11 Folia 1 iu& ni^ 46 41 U9-/ 11 IfS-I"----Mile IU& tTlST 4 40 -------------------------------------------------------------Hslc 111 11 110 11 116 11 111 11 114 11 ----------------------------------------------- __ tunaIt -- 111 111 110 350 360^' 11 51 11 11 41 bung Liver Utnun(|laiarfiCMi llApAtOlA* 11 3 1 11 42 1 1 1 1 3 Hawury Uml Ouci AilenocArclnoMUi SqiuMKKi* cell carciooM Aapl*tlc carcinoma {kHiaufc L<k>i*Ac2ll%_____________ Heaeniery lleaangUHiarccma_______________ ____________1_______1_______1_______1 _ _ 11 11 11 ________ l____________ _____________________l__ Kidney Adtnowa _______ _______________ I_________________________________________ _______________ Skin ao*c ant hoaaa_______________ _____________________________1_______________ ____________ i____________ _______ _______ _______________________________________________2_______2_______ -- ________________________i____________________________________________ ___________________ -- __________________________________________________ -- / Includes only alee thet were found-to hive tuaore during 10 to 11 nonthe exposure to VC or VUC b/ Severity of leelneei I - wild; 1 - noderite; 1 - lurked; * - severe; 1 - guestloneble. cl Oeetli or unscheduled retainer Ion. Ho tuaore were fouud In control alee during this period. CO tr* o oo w co to to SL 083924 TABLE 73 CUMULATIVE TUMOR INCIDENCE IH MICE PURIMG 12 MONTHS EXPOSURE TO VC OR VDC Control <f ? 50 ppm VC cT $ 250 ppm VC J? 1,000 ppm VC t% 55 ppm VDC d? Tumor Lung Bronchiolo alveolar adenoma Acinar proliferation Metastatic squamous cell carcinoma 1/26^ 0/36 0/26 0/36 8/29 1/29 0/26 0/36 0/29 4/34 1/34 10/29 3/29 12/34 1/34 22/33 0/33 26/37 2/87 2/34 0/29 2/34 0/33 8/37 6/35 2/35 0/35 0/35 0/35 0/35 Liver Hemang1osa rcoma Hepatoma Hepatic cell carcinoma 0/26 0/26 0/26 0/36 0/36 0/36 3/29 0/29 0/29 0/34 0/34 0/34 7/29 0/29 0/29 16/34 0/34 0/34 13/33 0/33 0/33 18/37 0/37 0/37 2/35 1/35 1/35 1/35 1/35 0/35 Mammary gland Duct adenocarcinoma Squamous cell carcinoma Anaplastic carcinoma Hemangiosarcoma 0/26 0/26 0/26 0/26 Mali gna n t 1 ymphouia 0/26 0/36 0/36 0/36 0/36 0/29 0/29 0/29 0/29 0/36 0/29 8/34 9/34 8/34 1/34 1/34 0/29 0/29 0/29 0/29 0/29 3/34 3/34 3/34 0/34 2/34 0/33 0/33 0/33 0/33 7/37 10/37 6/37 7/37 2/33 3/37 0/35 0/35 0/35 0/35 0/35 0/35 0/35 0/35 0/35 0/35 Hemangiosarcoma Heart Skeletal muscle f1 Gastrointestinal tract o00 Kidney oj Pancreas to Mesenteric lymph nodes Epididymus and testes Mesentery 0/26 0/26 0/26 0/26 0/26 0/26 0/26 0/26 0/36 0/36 0/36 0/36 0/36 0/36 0/36 0/36 0/29 0/29 0/29 0/29 0/29 0/29 1/29 4/29 0/34 0/34 0/34 0/34 0/34 0/34 0/34 1/34 1/29 0/29 0/29 0/29 0/29 0/29 0/29 1/29 0/34 1/34 1/34 0/34 1/34 0/34 0/34 0/34 0/33 0/33 0/33 0/33 0/33 0/33 0/33 0/33 0/37 0/37 0/37 1/37 0/37 1/37 0/37 0/37 0/35 0/35 0/35 0/35 0/35 0/35 0/35 2/35 0/35 0/35 0/35 0/35 0/35 0/35 0/35 0/35 ( c't tr* f- f m i fr> novh nntii ( TABLE 73 (Continued) CUMULATIVE TUMOR INCIDENCE IN MICE DURING 12 MONTHS EXPOSURE TO VC OR VDC Control 50 ppm VC 250 ppm VC o* 1.000 ppm VC 55 ppm VDC <S_ $ Hemangioma Mediastinum Connective tissue (adjacent to salivary gland) 0/26 0/26 Kidney Adenoma 0/26 Skin Keratoacanthoma 0/26 0/36 0/29 1/34 0/29 0/34 0/33 0/37 0/35 0/35 0/36 0/29 0/34 0/29 0/34 1/33 0/37 0/35 0/35 0/36 1/ 29 0/34 0/29 0/34 0/33 0/37 1/35 0/35 0/36 1/29 0/34 0/29 0/34 0/33 0/37 0/35 0/35 a/ Number of mice with turaore/total number of mice that died or were terminated during 12 months ex poaure to VC or VDC. SL 083925 \l l * > '* <. <1J ] TABLE 74 SUMMARY OP TISSUE LESIONS AND M/E RATIOS IN MICE AFTER EXPOSURE TO VC OR VDC FOR 1 MONTH Jl I l l SL 083926 / Severity of lesion: I * alld, 2 - Moderate, 3 - Marked, 4 - severe, j - questionable. TAItl.E Ti .ta_tUS tutu ei`hsiihe w vc tm vuc rim i mmins ? 6 8 ol */ Severity t lealoiu I - -Mil. 1 - lerl. 1 - -erked. 4 - aavere. t - 4ueat louabla, + - preaent. i a/ Lesion2' Sex Her No, FAULK 76 siihhahit ok tissue i.ksiuhs amp h/k maims in hick AFTKK KXI'OSUHK TO VC OH VPC FOH 3 MONTHS Hale Com lot 1_______________ __________________ VC A(li.d('H" )tl ppin)_____________________________VPC (55 ppeQ Fema 1 e _ Hu t! Fenia le Me e Feoiale 9 lfi 11 11 45 46 42 48 225 226 11Z 22s 261 262 263 m Hi 120 122 3011 333 m 335 Heart Focal Myocardial degeneration - - --- i11 L Hrouch|olo-atvcolar adenuaa Focal acinar epithelial proliferation 11 1 l.lver Hlcrofoci of nononticlear col la Intranuclear Indue Iona Focal necrosis Salivary Gland Focal chronic InfUMAtlon 11 + ++ - + -* + i 1 1 11 + i + + -t + + 1 1 + Kidney Focal, chronic InteratIt lal nephritis l 1 lubuJar caata 11 1 Hlcroca1cul1 1 1 1 1 1 11 1 1i1 1 Done Marrow H/K ratio 1.6 1.6 1.2 1.4 1.7 1.5 1.6 1.4 1.5 1.6 1.7 1.5 1.5 1.7 1.6 1.5 1.7 1.7 1.7 1.6 1.7 1.6 L o/ Severity of lealone; t - olid, 2 - luJame, 3 - outlied, 4 - severe, f - questionable, t - present. in tr* 6^80 CO TABLE 77 SUMMARY OF TISSUE LESIOMS (OTHER T11AH TUMORS) AND M/E RATIOS IN MICE EXPOSED TO VC OR VDC FOR 6 MONTHSa/ / Duel not include deaths and unscheduled tariatnatloue that occurred during the 4 to 6 Months, b/ Severity of leelon; l - olid; 2 - Moderate} 3 - Marked; 4 - severe; * - questionable. SL 083929 i i.t.i I TABLE 78 SUMMARY OF TISSUE LESIONS (OTHER THAN TUMORS) AND H/E RATIOS IN MICE EXPOSED TO VC OR VDC FOR 9 MONTHS--^ J.C. loll-^ VUC piiih) Halea Halea fcmrtleu ___Httjea _ Feou | ea __ 12 l( B 51 a S5 233 234 235 236 26ii 3Ui 306 307 30M 341 342 343 344 Heart HyucjirdlLta I1 Thickening of artarlal wall and krlarterlt la liUllg Lhrontc uirtne pitiuiuala _ _____ _ __ l.lv( I'artal InMamutton Hlcrofocl of aonomicluc col la Focal ileKenaratlon and nacroala Intranuclear Indus Iona _ Urjje 6aaojit)lllc_ouc______ Salivary Gland Chronic UflaHUtlon Cectita 1*1 nwoi hu) In Kidney Chronic Interstitial nephritic III I I 1 Tubular dilation and caste II 2I3 2 _ Cloneru lonephrK Adrenal Spindle cell Infiltration Largo cella with yellowish _ _ vacua la tl_njr foaia^ I2 Spleen l.ynqihold hyperplaala 2II Vacuo I teat ion __ txUaw<lulUrjrJieiutH()olMUa Heaentarlc l.ywpti llieauttiderosla Bone HurriM Setaer H/K ratio I I JiL 'L L .Li 1.5 1.5 1.4 1.1 1.9 1.1 1.1 1.5 / U>di uut Include death* or uairliidultd lamination* tliul occurred during 1 Co 9 Muutlia. b/ Severity of leelon; I - wild; 2 - Moderate; I - narked, 4 - eeverc; 1 - guuettunable. lit) 2 II 1.1 1.4 1.5 1.6 1.7 1.5 1.9 1.4 SL 083930 i*m 79 ftUHHMlT OF TjSSHE IKSIOHS fOTHKB THAIJ lUHtfflttl AHU H/t! HATIOS Ity MICK exposed fU VC ok vw: ym I 2_Ht m IISi 7 Cunt roll__________________ yn (SO m-lVPC Q5 pp) Hilo Fte-- lew Males_____ aicd _ 21 22 23 2 57 56 59 60 61 62 61 w li 56 91 9'. 95 309 110 111 112 111 214 215 316 145 346 347 J4U 149 350 351 352 leart jJygcnrditlA 22I Chronic aurJo* _ _ fneuiwaii _ _ _ _ _ Mvir Oi atended slnusolda Fucl n*cro*i Hicrofocl of Mono- nuclear celle Degeneration JntiiuucLr Inclu- * Juus 1 Cyat and auLecute latlanutloR __ _ __________ _ Satatl Intaatin* _ Aw(Ui^ali _3_ _ j Ccua __ fiRworw la kt_________________ 1-.A. _I 6* 11 vary Gland MUiofocI of Mono- ____nuc!* cHa ______ Klduey CbioRle lataratiCUl naphritla I1 3 lobular dilation and a* ____ciitt _________________ Ovary Cyat _ AMyloldoela___________ _ ___________ lit a rue Distended endoawtriel _ __ jlaikdi_________________________________ T-atia Tubular necroale and Mineralisation _ lajloldoill__________________________ idrtntl dpIndia call Infiltra tion Hononuclaar Inflitre - 31 t Ion _ iylol4oili___________________________ Spleen Amyloldoal* Lyuphold hyparplaaia 2 tatraaedul lery haMa _ _ topilaata ___________________________ M/E Ratio ~ 1.7 T.I iTi 1.7 1.6 1.6 1.9 1.6 2.2 I1 3 II 2 11 3 14 2 1 23 24 3 __ 32 1.7 1.6 1.6 1.6 1.7 1.6 1.6 1.6 1.9 1.6 2.1 -- 1.9 1.9 l.} 2.0 |. SL 083931 4/ "ijoen not Include" ifeulhe or unscheduled tarnations chat occurred during 7 to 9 Mootha. h/ Severity of Union* l - Mild; 2 - nodorota; 3 - marked, 4 - sewer*; 9lf lonahU. TABLE 80 DNA SYNTHESIS AND DNA CONCENTRATION OF LIVERS FRCM MALE MICE EXPOSED TO 50 PPM VC FOR II MONTHS VC (ppm) 0 50 DFM mg DNA 2886 + 240(8)2/ 4232 463(7)-- mg DNA g Liver 2.3 + 0.1 2.5 + 0.1 / Mean + S.E. (Number of observations). / Significantly different from control (two sample rank test^/). |c r l>|( 117 SL 083932 TABLE 81 HEPATIC ALA SVNTHETASE ACTIVITY IN RATS EXPOSED TO VC FOR 12 MONTHS VC Concentration 0 ppm 50 ppm 250 ppm a Moles/mg Proteln/hr Male Female 0.25 + 0.03^/ 0.20 + 0.01 0.26 + 0.04 0.22 + 0.02 0.24 + 0.03 a/ Mean + S.E. for three determinations. 118 SL 083933 TABLE 82 URINARY ALA CONCENTRATION IN RATS EXPOSED TO VC FOR 12 MONTHS VC (ppm) Sex Urine Volume (ml )--> Urinary ALA Cone entration^-' 0 50 250 1,000 Male Male Female Female Male Male Female Female Male Male Female Female Male Male Female 3 6 3 27 6 27 21 25 13 7 31 7 7 10 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 a/ Urine c<ollected overnight b/ Determined on a 0.,5 ml sample of urine and expressed on a scale of 1 (Oto 1 mg7. ALA) to 6 (6 to 10 mg7. ALA) with 2 equal to 1.0 to 1.5 mg7 ALA. 119 SL 083934 TABLE 83 Number of Rats in Dominant Lethal Study with Vinyl Chloride (VC) and Vinylidene Chloride (VDC) No. of Male Rats Tested Mated with 2 females Mated with 1 female Mated with 0 females Fertile^/ No. of Female Rats Mated Pregnant with 1 dead Implant with 2 dead implants with 3 dead implants Chamber Concentration (pom) VC VDC 0 50 250 1000 55 12 12 12 12 12 12 10 11 9 0 n* 1 2 2 0 0021 12 11 10 10 10 24 22 23 18 20 24 20 lefe/ 14b/ 13- 7 5753 3 2011 2 2102 a/ Fathered at least one litter with one or more viable Implants. 31 b/ Significantly different from control (Fisher' s exact probability test, p<0.05). 120 SL 83935 TABLE 84 Macing Results in Dominant Lethal Study with Vinyl Chloride (VC) and Vinylidene Chloride (VDC) Corpora lutea/dam Implants/dam Implants/corpora lutea x 100 Viable/total implants x 100 Chamber Concentration (oora) VC _ VDC 0 50 250 1000 55 15.8i0.62/ 14.30.8 15.210.3 15.1+0.6 15.110.5 13.7+0.8 12.0+0.9 13.4+0.5 11.9+0.9 14.210.5 86i3 83+5 884 816 94+2 922 923 93+3 952 9313 a/ Mean S.E, for the values calculated on a dam basis for the number of pregnant rats indicated in Table 83. 121 SL 083936 TABLE 85 LEVELS OF ALPHA-FETOPROTEIN ('AFP') IN RATS EXPOSED TO.. VC FOR 9. MONTHS Rat No, 17 18 19 20 53 54 55 56 89 90 91 92 125 126 127 128 161 162 163 164 197 198 199 233 234 235 236 269 270 272 Sex M M M it F F F F M M M M F F F F M M M M F F F M M M M F F F Exposure Gtoud Control Control Control Control Control Control Control Control 50 ppm VC 50 ppm VC 50 ppm VC 50 ppm VC 50 ppm VC 50 ppm VC 50 ppm VC 50 ppm VC 250 ppm VC 250 ppm VC 250 ppm VC 250 ppm VC 250 ppm VC 250 ppm VC 250 ppm VC 1,000 ppm VC 1,000 ppm VC 1,000 ppm VC 1,000 ppm VC 1,000 ppm VC 1,000 ppm VC 1,000 ppm VC ft OCM o V PR AFP/mi of ` <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 <0.0225/ <0.022 0.034 <0.022 <0.022 <0.022 <0.022 <0.022 <0.022 a/ Rat found to have hepatic hemangiosarcoma. 122 SL 083937 TABLE 86 LEVELS OF ALPHA-FETOPROTEIN (AFP) IN MICE EXPOSES TO VC OR 7DC Mouse No. 35 133 134 135 137 177 201 240 241 244 245 248 269 269A 270 272 273 274 275 277 278 324 Exposure Length of ;ex Grouc Exposure(weeks) M Control F 50 ppm VC F 50 ppm VC F 50 ppm VC F 50 ppm VC M 250 ppm VC F 250 ppm VC M 1,000 ppm VC M 1,000 ppm VC M 1,000 ppm VC M 1,000 ppm VC M 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC F 1,000 ppm VC M 55 ppm VDC 36 36 36 36 34 33 34 38 36 34 34 30 36 36 36 34 34 34 34 31 30 36 AFP/ml of Plasma <0.04 <0.04 <0.04 <0.04 <0.04 0.29-/ <0.045/ 3.96--/ <0.04i/ <0.04^-/ <0.04^/ <0.04 <0.045./ <0.04/ <0.045/ <0.04 <0.04 <0.045./ <0.045./ <0.04 <0.045./ 1.92-/ Mouse found to have hepatic hemangiosarcoma Mouse found to have a hepatoma. 123 SL 083938 SL 083939 Figure aha lation Chambers and Holding Racks. i i i.i ii ii Jt SL 083940 Figure 2 - Diagram of a Chamber Showing the Positions of Sampling Lines. SL 083941 Figure 3 - Average Weekly Analytical Concentrations and the Range of all Samples for VC. i 140 130 120 - Ii K> '-J 10 a_ _L 10 12 14 16 10 20 22 24 26 2a 30 32 34 36 38 40 42 44 46 40 50 52 WEEK Ftgurte 4 - Average Weekly Analytical Concentrations and the Range of all Samples for VDC. SL 083942 to 00 CO t* o0O0J kO ruf>* Figure 5 - Body Weights of Rats Exposed to VC or VDC. tvoO o CivPpD $> Cotiltol )I Figure 6 - Weight Gram of Mice Exposed to VC or VDC. APPENDIX___ I MANUAL FOR HEMATOLOGY, CLINICAL LABORATORY TESTS, HISTOPATHOLOGY, STATISTICAL ANALYSIS, AND NORMAL VALUES Cheng-Chun Lea Chuen-Bin Hong Jagdish C. Bhandari Judith D. Girvin John J. Kowalski Midwest Research Institute January 1977 SL O83945 TABLE OF CONTENTS PaSe I. Hematology and Clinical Laboratory Tests...................................................... 1 A. Hematology......................................................................................................... 1 B. Clinical Blood Chemistry........................................................................... 2 C. Urinalysis......................................................................................................... 3 D. Occult Blood in Feces .......................................................................... 4 E. Precision of Hematology and Clinical Blood Chemistry Tests............................................................................................................. 4 1. Reproducibility .......................................................................... 4 2. Reproducibility Within a Test Day........................................ 4 3. Proficiency Test Service..............................................................5 XI. Histopathology.............................................................................................................. 5 A. Necropsy and Gross Examination..............................................................5 B. Organ Weights ........................................................................................... 5 C. Tissues for Microscopic Examination ............................................ 6 D. Fixation and Staining of Tissues..........................................................6 III. Statistical Analysis................................................................................................ 6 IV. Normal Values ........................................................................................................ 7 A. Hematology, Clinical Laboratory Tests and Bone Marrow . . 7 B. Absolute and Relative Organ Weights ............................................ 7 C. Presence of Various Substances in the Urine ........................... 7 D. Occult Blood in Feces .......................................................................... 8 V. References...................................................................................................................... 8 Tables A - 0............................................................................................................................. 10-24 iii SL 083946 HEMATOLOGY. CLINICAL LABORATORY TESTS, HISTOPATHOLOGY, STATISTICAL ANALYSIS, AND NORMAL VALUES I. HEMATOLOGY AND CLINICAL LABORATORY TESTS The usual blood sample from dogs is 8 ml, from monkeys 4 ml, and from rats 0.3 ml for hematology and about 8 ml for full analysis at termination. A. Hematology The following hematological analyses are performed on all blood samples from rats, dogs and monkeys. 1. Erythrocyte and leukocyte counts: A Coulter Electronic Particle Counter with 100 4 aperture is used.A/ Particle-free diluents (Isoton for RBC, Zap-Oglobin in Isoton for WBC, Coulter Electronics, Inc.) are counted to establish the background. Each blood sample is counted in duplicate. For each test day, two control blood samples (Diagnostic Technology, Inc.) are counted separately In duplicate. 2. Hematocrit: Hematocrit is determined in capillary tubes using a microcapillary centrifuge (International Equipment Company, Model MB). Two control blood samples (Diagnostic Technology, Inc.) are measured separately in duplicate. 3. Hemoglobin: Hemoglobin is measured as cyanomethemoglobin.l/ Each blood sample is measured in duplicate. Cyanomethemoglobin (Coulter Electronics, Inc.) is used as the standard. For each assay, two levels of the standard are used and two control blood samples (Diagnostic Technology, Inc.) are measured in duplicate. 4. Methemoglobin (Met-Hb): Met-Hb is measured by the method of Dubowski.A/ A positive control is made by adding potassium ferricyanide to control blood.5 6 5. Heinz bodies: Heinz bodies are stained with methyl violet and the percent of Heinz bodies is calculated. 6. Mean corpuscular volume (MCV): MCV is calculated as follows: MCV (m^) - Hematocrit x 10 Erythrocytes in millions/mm^ 1 SL 083947 7. Mean corpuscular hemoglobin (MCHb): MCHb is calculated as follows: MCHb (mmg) Hemoglobin (gm %) x 10 Erythrocytes in millions /mm-' 8. Mean corpuscular hemoglobin concentration (MCHbC): MCHbC is calculated as follows: MCHbC (gm %) * Hemoglobin (gm 7.) x 100 Hematocrit 9. Differential leukocyte counts: Wright's stain is used to stain the leukocytes for examination. 10. Reticulocyte count: Reticulocytes are counted by the methy lene blue method using the Miller disc.it/ 11. Platelet count: A Coulter Electronic Particle Counter with 70 m aperture is used.2/Particle-free Isoton is used as diluent and counted to establish the background. At weekly intervals, platelets are also visually counted in a hemocytometer with a phase microscope for comparison.-^/ 12. Clotting time (dog and monkey): Clotting time is determined by the capillary tube procedure using two capillary tubes.!./ The time elapsed from the appearance of the blood from the animal and coagulation in either tube is measured. B. Clinical Blood Tests The following clinical blood chemistry tests performed on all blood samples from dogs and monkeys and on blood samp. 3 from rats at termination. 1. Blood glucose: Fasting blood glucose is determined by Stein's hexokinase method../ Standard glucose solution (Dade) is used to establish a standard curve. For each assay, one level of the standard and two con trols (Reference Serum, Worthington; and Validate, General Diagnostics) are measured. 2. Serum glutamic-oxaloacetic transaminase (SGOT): SGOT is measured by the method of Amador and Wacker.!/ Validate and Reference Serum are used as the enzyme reference for each assay. 2 SL 083948 3. Serum glutamic-pyruvic transaminase (SGPT) : SGPT is measured by the method of Henry et a1.12/ Validate and Reference Serum are used as the enzyme reference for each assay. 4. Alkaline phosphatase: Alkaline phosphatase is measured by the method of Bowers and McCcmb^XI/ Validate and Reference Serum are used as the enzyme reference for each assay. 5. BUN: BUN is measured using the BUN Strate Kit (General Diag nostic) which is based on the urease method.iZ/ Three levels of Calibrate (General Diagnostics) are used to establish a standard curve. For each assay, two controls (Calibrate I and Validate) are used as the reference. 6. Creatinine: Creatinine is measured by a modified kinetic alkaline picrate procedure.A2/ Creatinine Standard Solutions (Sigma Chemi cal Company) are used to establish a standard curve. For each assay, two levels of the standard and two controls (Calibrate I and Validate) are used as reference. 7. Lactate dehydrogenase (LDH): LDH is measured by the method of Wacker et al.A^/ Precinorm E and Frecipath E (Boehringer, Mannheim Corporation) are used as the enzyme controls for each assay. 8. q-Hydroxybutyrate dehydrogenase (a-HBDH): a HBDH is measured by the method of Rosalki and Wilkinson.--/' Precinorm E and Precipath E are used as the enzyme controls for each assay. 9. Creatine phosphoklnase (CPK): CPK is measured by the improved procedure of Rosalkii^/ base<T onTthe methods of Oliver.iZ/ Precinorm E and Precipath E are used as the enzyme controls for each assay. C. Urinalysis Urine samples are collected from animals before and during treat ment as are the blood samples. The urine from rats is collected by slight manipulation of their body, and samples within each group are pooled. The monkeys and dogs are placed individually in metabolism cages, and urine is collected in the stainless steel pan. The urine from each dog and the pooled urine from rats are tested and examined for the following: 1. Protein: Urinary protein is determined with Labstlx (Ames Company, Elkhart, Indiana). 2. Sugar: Urinary glucose and reducing substance are determined with Labstix (Ames Company). 3 SL 083949 3. Microscopic examination: Urine samples are centrifuged and the supernatant discarded. The residue is resuspended and examined micro scopically for the presence of erythrocytes, leukocytes, epithelial cells, and crystals under high power field and for casts under low power field. A positive urine control prepared with known amounts of protein and glucose in saline adjusted to pH 6.0 is run with each assay to check the reliability of the Labstix. fi D. Occult Blood in Feces Fecal samples are collected from animals before and during treatment as are the blood and urine samples. Occult blood in the feces is determined with Hematest Reagent Tablets (Ames Company, Elkhart, Indiana). A positive control (whole blood) and a negative control (distilled water) are included with each assay to check the reliability of the Hematest tablets. E. Precision of Hematology and Clinical Blood Chemistry Tests 1. Reproducibility For erythrocyte and leukocyte counts, hematocrit, hemoglobin, and the various clinical blood chemistry tests, the same control blood samples or control standards are used for day-to-day assays. The replication of results are excellent and are summarized in Table A. The determination of differential leukocyte counts and reticulocyte counts are performed by experienced personnel. At weekly intervals, a blood sample is counted by two or more personnel to confirm the accuracy of the counting. Also at weekly intervals, the platelet counts obtained from a Coulter Electronic Particle Counter are compared with the direct visual counts in a hemocytometer using a phase microscope. 2. Reproducibility Within a Test Day At monthly intervals, a blood sample is taken from a control dog and six or more determinations for erythrocyte, leukocyte, reticulocyte,and platelet counts, hemoglobin, and various clinical blood chemistry tests are performed to establish the reproducibility within an assay. The results are summarized in Table B. SL B^S0 4 3. Proficiency Test Service We subscribe to the Proficiency Test Service of the Institute for Clinical Science, Hahnemann Medical College, Philadelphia, Pennsylvania (F. Wm. Sunderman, M.D., Director). On the first day of each month, this service sends two samples containing two different sera or solutions to all subscribers for measurements of one or more of the parameters usually analyzed in clinical laboratories. Participants report their results on a form fur nished by the service. On the 15th day of the month, each participant re ceives a report from the service which includes: the results of a statistical analysis of the values reported by all the participating laboratories; a current review of pertinent methodology; a comprehensive bibliography; and validation of the results which the participating laboratory reported. This service enables each participating laboratory to obtain an unbiased and criti cal assessment of its proficiency in relation to that of 1,000 or so other clinical laboratories throughout the country. The service has been in con tinuous operation since 1949 and was given endorsement by the American Society of Clinical Pathologists in 1952 and by the Association of Clinical Scientists in 1957 and 1968. Our results have been found to be satisfactory and are summarized in Table C. II. HISTOPATHOLOGY A. Necroosy and Gross Examination At termination or prior to imminent death, rats are killed with ether, and dogs and monkeys with an overdose of sodium pentobarbital. Animals that die on tests are.kept refrigerated but not frozen until necropsy. The general physical condition and nutritional status of each animal at the time of death or termination are observed and recorded. Necropsy is performed as soon as possible after death. Gross changes of all tissues are carefully examined and recorded. B. Organ Weights The brain, liver, spleen, kidneys, adrenals, thyroids and gonads are trimmed free from surrounding tissues and weighed. The organ weight to body weight and/or brain weight ratios are then calculated. 5 SL 083951 C. Tissues for Microscopic Examination i Tissues to be examined include the eye, skin (breast), trachea, lung, tongue (except rat), salivary gland, liver, gallbladder (except rats), pancreas, esophagus, fundic and pyloric stomach, duodenum, jejunum, ileum, cecum, colon, kidneys, urinary bladder, gonads, and accessory organs, diaphragm and gracilis muscle, anterior pituitary, thyroids/parathyrolds, adrenals, tonsil (except rat), thymus, spleen, prescapular (except rats) and mesenteric lymph nodes, rib bone with bone marrow, brain (sagittal section for rats; coronal sections of cerebral cortex, cerebellum, and brain stem for dog and monkey), spinal cord (lumbosacral plexus, dog and monkey), sciatic nerve and any other structures not mentioned which show abnormal gross changes. D. Fixation and Staining of Tissues All tissues are cut not to exceed 1 cm in thickness for fixation. For most tissues, neutral buffered 10% formalin is used. Sufficient volume of fixing solution is used and the tissues are changed to a fresh solution after 24 hours. The fixed tissues are processed in an Autotechnicon for dehydration, clearing, and infiltration and then embedded in paraffin. Routine H & E staining is used to stain the sectioned tissues for micro scopic examination. Supplementary tissue fixatives and staining techniques may be employed for more positive identification of special lesions such as calci fication, pigments, fat deposition and other abnormal changes. HI. STATISTICAL ANALYSIS Data are analyzed statistically using the Dunnett's multiple com parison procedure following an analysis of variance,or our modification of this procedure for uneven numbers among groups. The chosen criterion significance is p < 0.05. The means of each group at various intervals during treatment are compared with pretreatment levels. For most experi ments in'beagles, three baseline (pretreatment) levels are obtained. The baseline levels for each animal are averaged and the mean is used in the analysis. In addition, the means of the various treated groups are compared with that of the control group at the respective time intervals. 6 SL 083952 IV. NORMAL VALUES A. Hematology, Clinical Laboratory Tests and Bone Marrow Since June 1971, we have used about 180 rhesus monkeys (Woodard Research Corporation, Herndon, Virginia, Primate Imports, Port Washington, New York, and PrimeLabs, Inc., Farmingdale, New Jersey) for various studies. The peripheral blood elements and clinical blood chemistry values of these monkeys before treatment and the myeloid/erythroid (M/E) ratio of the bone marrow of the monkeys used as normal controls varied among individual ani mals. The mean S.D. and the range of the various parameters for the males and females are summarized in Tables D and E, respectively. Since September 1971, we have used about 525, 5 to 9 months old, beagles dogs (AKC registered, Hazelton Research Animals, Inc.). The periph eral blood elements, clinical blood chemistry values and the M/E ratio of the bone marrow varied considerably among individual dogs. The mean + S.D. and the ranges of the various parameters for the males and females are summarized in Tables H and I, respectively. During the same period, we have used about 500, 7 to 10 weeks old, male albino rats (CD Strain, Charles River Breeding Laboratories). As for the dogs, the individual variations of the peripheral blood elements, clinical blood chemistry values and the M/E ratio of the bone marrow were large. The mean + S.D. and the ranges of the various parameters for these male rats are summarized in Table L. B. Absolute and Relative Organ Weights Organ weights, both absolute and relative to body weight, of rhesus monkeys, beagle dogs, and albino rats are summarized in Tables F and G, J and K, and M, respectively. These were control animals used be tween June 1971 and December 1976. C. Presence of Various Substances in the Urine Various substances occasionally occurred in the urine of monkeys, dogs and rats. The results are summarized in Table N. Large percentage of urine samples from monkeys contained epithelial cells, i.e., 34.7% to 52.0%. Other substances occurred in 8.1% or less of the urine samples. In dogs, protein, erythrocytes, leukocytes and epithelial cells were present in 19.1 to 21.6%, 16.5 to 19.8%, 22.6 to 24.6% or 24.7 to 25,7%, respectively, of the samples from dogs collected for analysis. Glucose, 7 SL 083953 crystals, and casts occurred in less than 22 of these samples. Some dogs had been bled and returned to the metabolism cages before the urine was removed for analysis. The high incidence of some of these substances in the urine of these dogs might be due to contamination with the fecal material and traces of blood dropped in the cage. Special care to avoid contamination has been undertaken. In rats, large percentage of urine samples contained protein, i.e., 29.8 to 36.02. A few samples contained erythrocytes, leukocytes, epithelial cells and crystals. D. Occult Blood in the Feces Less than 10% of the feces samples from monkeys or dogs was positive with the Hematest for occult blood. The results are summarized in Table 0. V. REFERENCES 1. Brecher, 0., M. Schneiderman, and C. Z. William: Evaluation of the Electronic Red Cell Counter. Am. J. Clin. Path., 2: 1439, 1956. 2. Selegson, D.: Standard Methods of Clinical Chemistry, Academic Press, Inc., New York, Vol. 2, p. 52, 1958. 3. Dubowski, K. M.: Measurement of Hemoglobin Derivatives in Hemoglobin, Its Precusors and Metabolites. (F. W. Sunderman and F. W. Sunderman, Jrs., eds.), J. B. Lippincott Company, Philadelphia, p. 29, 1964. 4. Brecher, G., and M. Schneiderman: A Time-Saving Device for the Count ing of Reticulocytes. Am. J. Clin. Path., ^20^: 1079, 1950. 5. Bull, B. S., M. A. Schneiderman, and G. Brecher: Platelet Counts With the Coulter Counter. Am. J. Clin. Path,, 44: 678-688, 1965. 6. Brecher, G., M. Schneiderman, and E. P. Cronkite: The Reproducibility and Constancy of the Platelet Count. Am. J. Clin. Path., 23: 15, 1953. 7. Hepler, 0. E.: Manual of Clinical Laboratory Methods, p. 83, Charles C. Thomas, Springfield, Illinois, 1935. 8. Slein, M. W.: Methods of Enzymatic Analysis (Bergmeyer, H. U., ed.), p. 117, Academic Press, New York, 1963. 8 SL 083954 T 9. Amador, E., and W. E. C. Wacker: Serum Glutamic-Oxaloacetic Trans aminase Activity: A New Modification and an Analytical Assessment of Current Assay Technics. Clin. Chem., j3: 343, 1962. 10. Henry, R. J., N. Chiamorl, 0. J. Golub, and S. Berkman: Revised Spectrophotometric Methods for the Determination of GlutamicOxaloacetic Transaminase, Glutamic-Pyruvic Transaminase, and Lactic Dehydrogenase. Am. J. Clin. Path., _34j 381, 1960. 11. Bowers, G. N,, Jr., and R. B. McComb: A Continuous Spectrophotometric Method for Measuring the Activity of Serum Alkaline Phosphatase. Clin. Chem., 12: 70, 1966. 12. Chaney, A. L., and E. P. Manback: Modified Reagents for Determination of Urea and Ammonia. Clin. Chem., 8: 130, 1962. 13. Lustgarten, J. A.: A Simple, Rapid, Kinetic Method for Creatinine Concentration. Clin. Chem., 18: 1419, 1972. 14. Wacker, W. E. C., D. D. Ulmer", and B. L. Vallee: Metalloenzymes and Myocardial Infarction. II. Malic and Lactic Dehydrogenase Activities and Zinc Concentrations in Serum. New Eng. J. Med., 225, 449, 1956. 15. Rosalki, S. B., and J. H. Wilkinson: Reduction of a-Ketobutyrate by Human Serum. Nature (London), 18, 1110, 1960. 16. Rosalki, J. B.: An Improved Procedure for Serum Creatine Phosphokinase Determination. J. Lab. Clin. Med., 69^, 696, 1967. 17. Oliver, I. T.: A Spectrophotometric Method for the Determination of Creatine Phosphokinase and Myokinase. Biochem. J., 61, 116, 1955. 18. Dunnett, C. W.: A Multiple Comparison Procedure for Comparing Several Treatments with a Control. J. Am. Stat. Assoc., 50^: 1096-1121, 1955. 9 SL 083955 TABLE A REPRODUCIBILITY AMONG TEST DAYS ON THE SAME CONTROL SAMPLES OR STANDARDS*/ No. of Determinations Mean + S. D. Range Erythrocytes (x 10/mnrb Normal level Abnormal level Hematocrit (vol %) Normal level Abnormal level Hemoglobin (gm %) Normal level Abnormal level Leukocyte Counts (x lQ^/mm^) Normal level Abnormal level Pasting Blood Glucose (mg Z) SGOT (IU/i) SGPT (IU//) Creatinine (mg %) BUN (mg %) Bilirubin (mg %) Alkaline Phosphatase (TQ/S) CPK LDH HBDH a/ Performed in December 1976. 20 20 20 20 20 20 20 20 20 23 23 18 19 11 22 11 8 8 4.51 + 0.07 2.32 0.04 44.3 0.40 22.8 0.60 14.2 0.20 7.4 0.20 7.3 + 0.50 17.6 0.80 163.0 + 7.5 61.7 3.9 51.3 2.6 2.2 + 0.3 9.8 0.2 0.8 0.1 71.6 + 5.4 153.0 7.7 98.0 2.4 226.0 7.2 4.36 - 4.67 2.25 - 2.40 44 - 45 22 - 24 13.6 - 14.5 6.9 - 7.8 6.8 - 8.7 16.3 - 18.7 151 - 178 55 - 68 46 - 55 1.6 - 2.6 9.5 - 10.2 0.8 - 1.0 62 - 80 139 - 161 95 - 101 214 - 238 SL 083956 10 TABLE B REPRODUCIBILITY WITHIN A TEST DAY ON THE SAME SPECIMEN!? Mean S.D.-^ Range Erythrocytes (x lO^/ranP) Reticulocytes (%) Hematocrit (vol %) Hemoglobin (gm %) Platelets (x 105/mm3) Leukocytes (x lO^/mnP) Bands (%) Neutrophils (%) Lymphocytes (%) Eosinophils (%) Basophils (%) Monocytes (%) Atypical (%) Nucleated RBC (%) Methemoglobin (gm Z) Pasting Glucose (mg SGOT (IU/;). SGPT (IU/i) Creatinine (mg %) BUN (mg %) Alkaline Phosphatase (IU/i) CPK LDH HBDH 5.90 + 0.14 0.63 + 0.12 46.8 + 0.6 16.1 0.2 1.56 0.07 10.8 0.4 00 64.3 3.1 29.0 4.9 3.2 0.8 00 3.4 0.9 00 00 00 96.7 + 3.0 23.2 2.8 25.3 + 2.1 0.6 0.1 9.0 0.0 63.5 1.1 44.0 1.6 38.5 + 1.6 42.0 + 1.6 5.73 - 6.08 0.44 - 0.79 46.0 - 47.5 15.8 - 16.1 1.49 - 1.66 10.2 - 11.3 0-0 61 - 69 23 - 35 2-4 0-0 3-5 0-0 0-0 0-0 32 - 101 21 - 28 24 - 28 0.5 - 0.6 9-9 62 - 65 43 - 46 37 - 40 40 - 43 aj Performed in October 1976. b/ Six determinations from an adult beagle blood sample. 11 SL 083957 TABLE C PROFICIENCY TEST SERVICE (PTS) REPORTS (1975-1976)-^ l Unknowns MRI Results PTS Results Participating Laboratories (10-90 Percentiles) Median Mean Acceptable Performance^/ Hemoglobin 13.8 gm % 13.1 gm Z 13.8 17.9 13.8 17.9 13.8 17.8 13.6 - 14.0 17.6 - 18.2 Serum Protein 6.6 mg Z 7.1 7.0 7.0 6.7 - 7.3 Fasting Glucose 272.0 mg Z 229.0 mg Z 264.5 221.4 266.0 220.5 263.0 222.5 240 - 290 200 - 240 BUN 12.1 mg % 12.0 12.0 12,2 11.0 - 13.0 38.4 mg Z 40.1 40.3 39.2 36.0 - 44.0 Creatinine 1.0 mg Z 4.3 mg Z 1.0 4.4 1.0 1.0 4.5 4.4 0.8 - 1.3 3.9 - 4.9 Bilirubin 3.9 mg Z 1.3 mg Z 4.16 1.78 4.15 1.80 4.14 1.77 3.5 - 4.6 1.5 - 2.1 Cholesterol 175.0 mg % 100.0 mg Z 161.4 109.8 161.0 109.4 162.0 111.0 145 - 175 98 - 120 Ca 15.7 meq/i 15.4 15.4 15.3 14.1 - 16.4 9.5 meq/i 9.8 9.8 9.8 9.2 - 10.3 Na 156.0 meq/i 155.8 156.0 155.5 153 - 158 K 7.3 meq/Jt 7.5 7.5 7.5 7.3 - 7.7 Cl 96.0 meqH 97.8 98.0 97.5 96 - 101 78.0 meq/i 79.4 79.0 80.0 77 - 83 Mg 1.0 meq/j? 1.1 1.1 1.2 0.9 - 1.4 1.9 meq/i 2.0 2.0 2.1 1.8 - 2.3 a/ To date, we have received unknowns for phosphorus, uric acid, and serum iron. We do not routinely perform these determinations, b/ Based on values submitted by participants by 10th of month. 12 SL 083958 IABI.E IIEHATOl.OdY. CMH1CAI, BI.OOD CHEMISTRY VA1.UES, AMD WINE HAKllOU (HYEtOID/KKYTlIlUHD) HATIOS OF HAI-E RHESUS hOHKKYSa f Erythrocytes (> 10*Vmm3) ReiIculocytea (Z) llenutocrlt (wol X) IkMOglobln (g Z) HCV (M3) HCIlb (MMg) HCIIliC (>g X) Platelets (x 103/n3) Leukocytes (x 103/mm3) Neutrophils 1 (Z> Neutrophils H (I) Lynphocytes (X) Eosinophils (X) Houophlls (X) Basophils (X) Atypical cells (X) Nucleated BBC (X) Fasting Clocose (ug X) SCOT (IU/#> SCPt (IU//) . Alkaline Phosphatase (lU/f) BUN (Mg X) Protli. T 1m (aec) SeruM Creat. (Mg X) Bilirubin Total (mg X) Direct (ug X) BSP 15 Min <Z ret.) Na <Eq/n K (mF.i|/() Cl (nEq/#) Ca (nEq//) Mg (nEq//) Bone Marrow Hycloid/erythrold ratio Ka | e_ Rheaus Honkeya Nuaiber Body Weight (kg) Stud ted Heap S.II. ioa 3. 74 t 0.50 ION 1. 74 1 0.50 ION 1.76 1 0.50 ma 1.76 0.50 108 1.76 0.50 108 3.76 J 0.50 108 3.76 4 0.50 99 3.76 1 0.50 108 3.76 i 0.50 108 3.76 1 0.50 108 3.76 t 0.50 108 3.76 i 0.50 108 3.76 0. 50 108 3.76 0.50 108 3.76 0.50 108 3.76 t 0.50 3.76 t 0.50 100 3.76 0.51 100 3.76 i 0.51 100 3.76 t 0.51 1(10 3.76 i 0.51 100 3.76 t 0.51 62 3.91 i 0.66 100 3.76 t 0.51 62 3.91 0.66 62 3.91 1 0.66 62 3.91 1 0.66 62 3.91 t 0.66 62 3.91 t 0.66 62 3.91 t 0.66 62 3.91 t 0.66 62 3.91 t 0.66 15 3.65 0.61 a/ llata collected between Juno 19/1 ami Deceober 1976. Observed Hesid ts Heun i S.l>. FLmge 5.51 * 0.6 5 0.97 1 0.U2 61.0 i 2.6 11.6 i 0.8 77.8 t 7.0 26.6 i 1.8 31 .6 1 . 3 1.08 0.65 10.6 6.9 0.18 0.65 39.30 17.72 56.83 17.76 1.91 1 2.62 1.37 i 1.58 0.06 * 0.20 0.00 l 0.00 0.00 0.00 96.9 t 15.2 31.7 1 9.2 11.1 i 7.8 360.0 1 116.0 19.5 1 7.5 10.2 t 0.7 1.1 t 0.3 1.75 6.61 0.07 - 2.61 17.0 - 50.0 10.8 - 15.6 69.6 - 117.3 21.0 - 11.6 27.2 - 36.1 0.80 - 7.10 1.8 - 10.1 0-2 10 - 81 13 - 86 0 - 11 0-7 0- 2 0- 0 0_0 59 - 127 20 - 60 15 - 66 163 - 501 12 - 65 9.3 - 11.9 0.6 - 1.8 0.1 t 0.2 0.0 i 0.0 18.0 t 7.6 156.0 1 19.1 6.8 1 0.6 109.0 * 6.6 5.2 1 0.6 1.6 t 0.1 0.0 - 0.8 0.0 - 0.0 2 - 36 166 - 179 3.9 - 5.7 91 - 11B 6.2 - 6.3 1.2 - 1.8 1.5 0.1 1.5 - 2.1 TABLE E HEMATOLOGY. C1.IHICAI. BLOOD CHKH1STBY VALUES. AND BONE MAHJtOH (MYEUHD/EBYTIIKOln) RATtOS OF FLHA1-K RHESUS MONKEYS-nf Feaa1e Nhesus Monkeys Nnnher Studied Rady Weight (V Hean * S.l). Erythrocytea (a IO*/m3> Ret Iculocytea (X) lleaatocrit (vol X) Heaoglobln (ga X) HCV (M1) MCIlb (Mpg) IlCllbC (ag X> Platelets (x loVaa*) Leukocytes (x loVaa^) Neutrophils I (X) Neutrophils M (X) Lyapliocytes (X) Eosinophils (X) Honophila (X) Basophils (X) Atypical cells (X) Nucleated NBC (X) Fasting Glucose (ag X) SCOT (IU//) SKPT (IU/#) Alkaline Phosphatase (lU/f) BUN (ag X) Froth. Tine (sec) Serna Crest, (ag X) Bilirubin Total (ag X) Direct (ag X) BSP 15 aln (X ret.) Na (oq/f) JC (mEq/t) Cl (aEq//) Ca (aEq//) Mg (oEq//) Bone Marrow Hyelold/erythrold ratio HI 81 81 81 81 81 HI 81 81 81 81 81 81 8) 81 81 76 HI 81 81 81 81 59 81 81 at 59 59 59 59 59 59 11 1.51 i 0.48 1.51 t 0.48 1.51 1 0.48 1.51 t 0.48 3.51 t 0. 48 1. 51 t 0.48 1.51 i 0.48 1. 51 f 0.48 1.51 i 0.48 1.51 1 Q.4B 1.51 i 0.48 1.51 i 0.48 3.51 t 0.48 1.51 t 0.48 1.51 i 0.48 1.51 t 0.48 1.56 t 0.50 1.51 0.48 1.51 t 0.48 3.51 t 0.48 3.51 t 0.48 1.51 t 0.48 1.56 t 0.41 3.51 1 0.48 1.51 1 0.48 3.51 1 0.48 1.56 1 0.41 1.56 1 0.41 3.56 4 0.41 1.56 i 0.41 1.56 0.41 1.56 t 0.41 1.49 t 0.62 Observed Results Me an * S.D. Ran 5. 11 * 0.40 1.07 t 0.54 41.5 J 2.8 11.1 i 1.0 77.7 * 5.1 24.6 i 1.7 11.6 * 1.4 3.11 i 1.21 9.5 i 1.9 0.10 l 0.41 16.41 t 11.12 60.18 i 13.26 2.28 t 1.10 0.75 t 0.98 0.05 t 0.22 0.00 0.00 0.00 1 0.00 92.1 i 15.1 12.1 i 7.6 10.1 i 7.6 349.9 t 112.3 17.3 1 4.2 10.5 t 0.9 1.1 t 0.1 0.1 t 0.1 0.0 1 0.0 16.4 i 8.1 158.2 t 6.5 4.8 t 0.7 109.0 t 6.1 5.1 * 0.5 1.6 t 0.2 4.25 _ 6.01 0.15 ~ 1.11 10.0 - 46.0 7.9 - 14.1 66.5 - 95.2 17.6 - 29.7 26.6 - 14.2 1.85 - 7.90 1.2 - 24.8 0- 3 13 * 56 41 - 79 0 - 18 0-4 0- 1 0- 0 0-0 57 - 116 20 - 70 12 - 19 148 - 57 2 11 - 29 9.7 - 12.1 0.6 - 1.7 0.0 - 0.8 0.0 - 0.0 5 - 14 147 - 174 1.9 - 6.2 95 - 111 4.3 - 6.1 1.1 2.0 1.4 t 0.1 1.0 - I.B SL 083960 a/ Data collected between June ll?l and Decenber 1976. -t- * TABLE F ABSOLUTE AND RELATIVE ORGAN WEIGHTS OF MALE RHESUS MONKEYS*/ Organ Weight Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (gm) Thyroids (gm) Testes (gm) Mean t S.D, 82 17 4.6 1.8 15.1 i 3.8 0.73 0.15 0.57 1.30 1.29 0.67 Absolute Range 64 - 122 2.0 - 9.3 8.0 - 22.0 0.45 - 0.86 0.37 - 0.81 0.53 - 3.30 Relative (per kg body weight) Mean S.D. Range Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (mg) Thyroids (mg) Testes (gm) 23.4 2.5 1.25 0.47 4.13 0.92 201 44 154 42 0.34 0.11 18.8 - 30.4 0.57 - 2.38 2.20 - 6.43 129 - 254 86 - 250 0.18 - 0.53 a/ Data collected between,September 1971 and December 1976 from 17 monkeys weighing 3.71 0.48 kg, used as control animals. SL 083961 15 TABLE G ABSOLUTE AND RELATIVE ORGAN WEIGHTS OF FEMALE RHESUS MONKEYS^/ Organ Weight Mean t S.D. Absolute Range Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (gm) Thyroids (gm) Ovaries (gm) 83 17 3.8 t 1.4 14.5 2.8 0.68 0.16 0.60 0.20 0.28 0.10 64 - 122 2.0 - 6.0 11.0 - 20.0 0.53 - 1.14 0.37 - 1.11 0.14 - 0.45 Relative (per kg body weight) Mean S.D. Range Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (mg) Thyroids (mg) Ovaries (mg) 25.4 5.8 1.16 0.49 4.40 0.86 212 80 173 66 82 t 28 19.2 - 37.4 0.60 - 1.89 3.20 - 6.25 138 - 438 97 - 346 43 - 140 a/ Data collected between September 1971 and December 1976 from 11 ~ monkeys weighing 3.39 0.58 kg, used as controls. 16 SL 083962 it ii TABLE H HEMATOLOGY, CLINICAL BLOOD CHEMISTRY VALUES. AMD BONE MARROW TmYELOID/ERYTHROID) RATIOS OP MALE BEAGLE DOGS5./ Number S tud led Erythrocytes (x 10**/ram3) Reticulocytes (X) Hematocrit (vol X) Hemoglobin (gm X) MCV (p3) MCHb ( ppg) MCHbC (mg X) Platelets (x 103/mm3) Leukocytes (x 103/mni3) Neutrophils I (X) Neutrophils M (X) Lymphocytes (X) Eosinophils (X) Monophila (X) Basophils (X) Atypical cells (X) Nucleated RBC (X) Fasting Glucose (mg X) SGOT (lU/f) SGPT (IU/f) Alkaline Phosphatase (IU/7) BUN (mg X) Bone Marrow Myelold/erythroid ratio 276 284 276 276 276 276 276 270 284 284 284 284 284 284 284 284 284 284 276 276 276 284 34 Male Beagle Doga Age Body Weight (kg) (months) Mean S.D. 4_7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4- 7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 8.3 + 1.7 8.3 1.7 8.3 1.7 8.3 1.7 8.3 + 1.7 8.3 + 1.7 8.3 + 1.7 8.4 + 1.7 8.3 + 1.7 8.3 + 1.7 8.3 + 1.7 8.3 1.7 8.3 1.7 8.3 + 1.7 8.3 + 1.7 8.3 + 1.7 8.3 + 1.7 8.3 + 1.7 8.3 + 1.7 8.3 1.7 8.3 + 1.7 8.3 + 1.7 59 9.4 + 1.6 Observed Results Mean + S.D. Range 5.55 + 0.73 0.72 + 0.46 41.6 + 3.5 13.5 + 1.4 75.6 + 8.3 24.6 + 3.0 32.5 + 1.5 2.91 + 1.02 11.9 + 3.5 0.55 + 1.06 56.81 + 9.47 37.94 9.26 2.76 2.93 1.78 1.84 0.01 0.10 0.11 + 0.37 0.02 + 0.10 100.9 12.6 23.2 + 7.4 25.7 + 7.9 73.3 + 18.5 12.1 3.3 3.62 _ 7.60 0.04 - 4.35 31 - 50 10.0 - 16.9 56.7 - 127.1 17.1 - 41.7 28.1 - 40.3 0.93 - 6.35 4.6 - 24.6 0-6 22 - 80 13 - 71 0 - 16 0 - 11 0- 2 0- 2 0-2 66 - 134 11 - 59 8 - 46 21 - 133 4 - 23 1.6 + 0.4 1.1 _ 3.0 aj Data collected between September 1971 and December 1976. SL 083963 TABLE I HEMATOLOGY. CLINICAL BLOOD CHEMISTRY VALUES, AND BONE MARROW (MYELOID/ERYTHROID) RATIOS OF FEMALE BEAGLE DOGSg/ Number Studied Female Beagle Dogs Age Body Weight (kg) (months) Mean S.D. Erythrocytes (x 10*Vra3) 257 Reticulocytes (X) 265 Hematocrit (vol X) 257 Hemoglobin (gm %) 257 MCV (fi3) 257 MCHb (mMS) MCHbC (mg X) 257 257 Platelets (x 10^/mm3) 227 Leukocytes (x 103/mm3) 265 00 Neutrophils I (X) 265 Neutrophils M (X) 265 Lymphocytes (X) 265 Eosinophils (X) 265 Monophils (X) 265 Basophils (X) 265 Atypical cells (X) 265 Nucleated RBC (X) 265 Fasting Glucose (mg X) 248 SGOT (IV/l) SGPT (IU/7) 257 257 Alkaline Phosphatase (IU/jf) 257 BUN (mg X) 265 Bone Marrow Myeloid/erythroid ratio 34 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 4-7 5-9 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 + 1.3 6.9 1.3 6.9 + 1.3 6.9 + 1.3 6.9 i 1.3 6.9 + 1.3 6.9 1.3 6.9 1.3 6.9 1.3 6.9 1.3 6.9 1.3 6.9 1.3 7.8 1.4 a/ Data collected between September 1971 and December 1976 Observed Results Mean + S.D. Range 5.59 + 0.73 0.74 + 0.52 42.3 + 3.5 13.7 + 1.3 76.7 + 9.7 24.8 + 3.3 32.3 + 1.6 3.08 1.15 10.9 3.4 0.54 1.16 57.08 10.10 37.15 10.46 2.37 2.25 1.94 2.01 0.01 + 0.09 0.11 0.43 0.03 + 0.17 99.6 14.4 23.5 7.2 25.3 7.0 73.5 19.2 12.4 + 3.3 3.27 - 7.75 0.04 - 5.05 32 - 51 11.0 - 18.6 55.8 - 128.4 17.1 - 41.6 28.7 - 40.4 1.08 - 7.95 3.8 - 26.9 0-7 31 - 85 10 - 61 0 - 13 0-9 0-1 0 -4 0-2 55 - 130 6 - 52 8 - 49 30 - 146 4 - 26 1.4 0.3 1.1 - 2.4 SL 083964 5 TABLE J ABSOLUTE AND RELATIVE ORGAN WEIGHTS OF MALE BEAGLE DOGS^/ Organ Weight _______________ Absolute Mean S.D. Range Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (gm) Thyroids (gm) Testes (gm) 264 51 58 25 53 10 1.12 i 0.26 1.03 0.32 6.60 t 4.56 166 - 384 22 - 167 32 - 71 0.74 - 1.75 0.55 - 2.50 1.32 - 18.00 Relative (per kg body weight) Mean S.D. Range Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (mg) Thyroids (mg) Testes (gm) 27.9 4.2 6.0 2.0 5.6 0.8 117 25 108 34 0.67 0.39 19.6 - 42.3 2.8 - 12.5 4.0 - 7.7 70 - 165 56 - 211 0.13 - 1.67 a/ Data collected between September 1971 and December 1976 from 51 dogs, weighing 9.3 1.8 kg, used as control animals. 19 083965 SL> *V TABLE L HEMATOLOGY. CLIHICAL BLOOD CHEMISTRY VALUES, AND BONE MARROW (MYELOID/ERYTHROID) RATIOS OF MALE ALBINO RATSS? Humber Studied Erythrocytes (x 10*VmnP) Reticulocytes (%) Hematocrit (vol Z) Hemoglobin (gm Z) MCV (fP) MCHb (wig) MCHbC (mg %) Platelets (x lO'VmnP) Leukocytes (x IO^/tiwP) Neutrophils I (%) Neutrophils M (Z) Lymphocytes (%) Eosinophils (%) Monophils (Z) Basophils (Z) Atypical cells (Z) Nucleated RBC (Z) Fasting Glucose (mg Z) SGOT (IU//) SGPT (IUfi) Alkaline Phosphatase (III//) BUN (mg Z) Bone Marrow Myeloid/erythroid ratio 527 461 525 525 525 525 525 473 448 448 448 448 448 448 448 448 448 125 125 125 125 125 109 Male Rats Age Body Height (gm) (weeks) Mean + S.D. 5.7 5-7 5-7 5-7 5-7 5-7 5-7 5-7 5-7 5-7 5-7 5-7 5- 7 5-7 5-7 5-7 5-7 10 - 12 10 - 12 10 - 12 10 - 12 10 - 12 168 + 22 168 + 22 168 22 168 + 22 168 + 22 168 + 22 164 + 24 164 + 24 164 24 164 + 24 164 + 24 164 + 24 164 + 24 164 + 24 164 + 24 164 + 24 348 72 348 72 348 + 72 348 + 72 348 72 10 - 12 349 + 63 a/ Data collected between September 1971 and December 1976 Observed Results Mean S.D. Range 5.84 0.54 3.04 1.80 45.1 + 3.2 13.7 + 0.9 78.1 + 16.3 23.7 + 2.6 30.5 + 1.8 4.93 1.23 15.4 + 4.0 0.07 + 0.31 14.1 + 6.2 83.63 + 6.75 0.64 + 0.91 1.23 + 1.73 0.01 0.15 0.01 0.12 0.10 + 0.42 130.9 i 17.2 108.2 + 34.5 34.2 + 16.5 94.9 30.0 16.4 + 4.7 3.24 -- 7.60 0.30 - 6.83 40 " 58 11.8 - 17.1 62.3 - 104.6 19.2 - 41.0 21.1 - 36.9 2.30 - 7.95 6.3 - 20.8 03 4 - 29 52 - 96 0-6 0 - 13 02 0-2 0- 4 94 - 165 63 - 223 17 - 120 32 - 153 8 41 1.7 + 0.5 1.0 -- 2.6 SL 083966 TABLE M ABSOLUTE AND RELATIVE ORGAN WEIGHTS 07 MALE ALBINO RATS*/ 5 Organ Weight Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (mg) Thyroids (mg) Testes (gm) Mean S.D. Absolute Range 10.89 2.87 0.65 0.11 2.64 0.37 63.6 9.5 26.3 5.8 2.98 0.51 7.18 - 15.09 0.34 - 0.89 1.84 - 3.58 21.9 - 73.5 14.3 - 37.7 1.76 - 3.81 Relative (per 100 gm body weight) Mean S.D. Range Liver (gm) Spleen (gm) Kidneys (gm) Adrenals (mg) Thyroids (mg) Testes (gm) 2.96 0.42 0.19 0.08 0.76 0.10 18.6 5.8 7.6 2.7 0.87 0.15 2.09 - 4.01 0.10 - 0.30 0.22 - 0.88 5.8 - 22.4 4.2 - 12.7 0.23 - 1.09 a/ Data collected between September 1971 and December 1976 from 139 rats, weighing 352 59 gm, used as control animals. 22 SL 083967 TABLE N PRESENCE OF VARIOUS SUBSTANCES IN THE URINE OF MALE AND FEMALE MONKEYS. DOGS AND MALE: RATS Species: No. of Animals: No. of-Collections: Glucose: < 250 mg X > 250 mg X 141--^ 141 Monkeys 18 98 qb! 2.0 (2) 00 615^/ 615 Dors 112 / 565*^ 0.2 (1) 0.5 (3) 0.7 (4) 0.2 (1) 84k/ 84 Rats a/ 18 56p 00 00 Protein: < 100 mg X > 100 mg X 3.5 (5) 0 6.1 (6) 2.0 (2) 19.3 (119) 2.3 (14) 17.3 (98) 1.8 (10) 29.8 (25) 0 36.0 (18) 0 RBC: -- Mod era te Excessive 1.4 (2) 0 3.1 (3) 0 16.4 (101) 3.4 (21) 13.3 (75) 3.2 (18) 3.6 (3) 0 8.0 (4) 0 WBC:--^ Moderate Excessive 1.4 (2) 0 2.0 (2) 0 18.7 (115) 3.9 (24) 20.9 (118) 3.7 (21) 0 0 4.0 (2) 0 Epithelium Moderate Excessive Crystal:--^ Moderate Excessive 31.2 (44) 3.5 (5) 0.7 (1) 0 44.9 (44) 7.1 (7) 2.0 (2) 0 21.0 (129) 4.7 (29) 0.2 (1) 0.2 (1) 21.9 (124) 2.8 (16) 0.7 (4) 0.7 (4) 0 0 0 0 8.0 (4) 0 2.0 (1) 2.0 (1) Casts: Positive 0.7 (1) 5.1 (5) 0 0.9 (5) 0 0 aj Pooled sample of 4-20 rats. b/ Baseline data collected from all animals employed between September 1971 and December 1976. c/ Data collected at weekly intervals for 4-7 collections from controls employed between September 1971 and December 1976. d/ Data collected at 2-week intervals for 2-4 collections from control rats employed between September 1971 and December 1976. e/ Percent of total (number of samples). f/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 cells/field (x 440). / Normal, 5 or less cells; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). h/ Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/field (x 100). TABLE 0 PRESENCE OF iOCCULT BLOOD IN THE FECES OF MALE AND FEMALE MONKEYS AND DOGS ' Species; No. of Animals: No. of Collections: 44/ 44 Monkeys 3 48b/ 118^ 118 Dogs 30 156fe/ Occult Blood: Negative 90.9 (40)SJ 95.8 (46) 94.1 (111) 91.7 (143) Positive 9.1 (4) 4.2 (2) 5.9 (7) 8.3 (13) aj Baseline data collected from all animals employed between July 1974 and December 1976. b/ Data collected at weekly intervals for 4-7 collections from controls employed between July 1974 and December 1976. c/ Percent of total (number of samples). 24 SL 083969