Document B53kvmZ1KOwLZwpb1beBq3v0j

I I I I I I I I I I I I I I I I AR226-2745 EFFECT OF FLUOROCARBON DISPERSING AGENTS ON THE LIVERS OF RATS AND DOGS Medical Research Project Nos Report No. 123-65 m* * powerful surfactants. In acute oral toxicity studies witlfthese compounds, the most striking abnormality in animals surviving a single sublethal dose was gross li ver A similar but less marked effect was observed In rabbits thaf^had a b s o r b e d ^ t h r o u g h the skin. Although liver enlargement is,.not an uncommon 'finding in chemical intoxications, the effect of H P ^ w a s remarkable in that 56 days after a single dose of 12 mg/kg thhe liver was approximately three times larger than normal. The experiments reported here were preliminary studies designed to determine more details concerning: ( 1) the histological changes, that occur with increasing periods of time_in the livers of rats after a single oral dose o f 8 f g ( )2 the biochemical changes during the period of rapid growth qnd recovery in the livers of rats treated w ith6 i f (3) the combinedi effects of oral administration of ethanol and jg||{in rat; (4) the effect of fiflij[upon the pentobarbital sleeping time in the r<at; and (5) the effect on liver fimction in dogs treated with ___ jin order to find some clin- 'ical lffloratory procedure that might be useful in detecting liver injury in personnel exposed to fluorocarbon dispersing agents. I. HISTOLOGICAL CHANGES DURING THE TIME OF RAPID GROWTH A. Prqced'ire; In order to study the changes in sze and structure of tr -"Tiver following a single oral dose ofjlHtJ 14 male rats were each iven a dose of 12 mg/kg. A second group of animals of the same age and weight served as controls. One test and one control animal were killed at Intervals over a period of 56 days and two test and two control animals at 75 and 128 days. The livers were removed, weighed and a section taken for microscopic examination. Company Sanitized. Does not contain TSCA CBi 2 B. Results: The rapid growth of the liver, in cerins of both weight and per cent of whole body mass, is shown In Figure 1. The increase in size was most marked during the first ten days, reached a maximum between 40 and 60 days, and then began to decrease in size, although some enlargement was still evident after 128 days. Morphological changes in the liver were evident 24 hours after the single dose. The initial change was characterized by markedly enhanced mitosis of the hepatocytes. This was less evident by the second day; by the ninth day, the mitotic activity had decreased. After the j6th day, most of the features which characterized the response were less intense. The structural changes in the cells were quite evident upon microscopic examination and appeared to result principally from an enlargement of the individual cells. I I . BIOCHEMICAL AND METABOLIC CHANGES IN RAT LIVER 1. Changes in the Liver of Intact Animals _ Procedure: Thirty male ChR-CD rats were given 12 rng/kg 111 i i 111|_11 oral dose. Two to three weeks later, eight of the"rats were sacrificed by decapitation, the livers removed, weighed, and analyzed. An equal number of untreated controls were examined to establish the normal range of valugs Jthat might be expected in rats of this age and strain. Twojj^jtreated rats and two controls were also sacrificed for microscopic examinationpf __ the tissue. Eight to ten weeks after the single oral dose o f T W l j a second group of ten rats was sacrificed and the remaining rats three months later. Tissue slices were prepared from a portion of the median lobe for respiration and choline oxidase measurements in a Warburg respirometer. One portion of the left lateral lobe was homogenized for measurement of alkaline phos phatase activity (APase), esterase activity, and the nucleic acids. Another portion was used for glycogen determination. The remain- . ing tissue was used to measure water, protein, fat, ash, chol esterol, phospholipids, and potassium. An average value was calculated f**r each group of animals and compared with the controls. Differences oetween groups were tested for significance with the "t" test. ' B. Results: The results of the biochemical measurements are given in Taoie l. Two weeks after the single dose o i T i w Q the livers of the treated rats were quite large in size, I S r e t h a n twice that of the controls, and comprised approximately 9# of the body weight. An increase in the alkaline phosphatase activity and phospholipids and a decrease in glycogen were the most important changes. Small but statistically significant (p^O.O^) increases in oxygen consumption, non-protein nitrogen, water and protein and decreases in the respiratory quotient, choline oxidase, RNA and DNA occurred. Two months after the single oral dose ofM B S , \ the livers were ao longer growing at an accelerated rate, forHrhei*B was no further increase in the absolute size of the liver and its portion relative I ! I I I I I B I 4 I I I I I I I - 3- to the whole body had decreased. The increase in phospholipids, protein and non-protein nitrogen, and decrease in glycogen respiratory quotient, RNA and DNA again was observed. Five and one-half months after the dose oftflHl]j the livers of the treated rats were still slightly larger than~normal, but the small differences in the biochemical measurements were no longer significant. These results indicate thatMBPjproduces significant bio chemical as well as morphologicalrciianges in the liver of rats. A number of these changes point to disturbances in the normal meta bolic and synthetic functions of the liver. The lowered concen tration of nucleic acids is consistent with the observed arrested mitotic activity. These effects are most evident when the liver is rapidly increasing in size. Later, when the accelerated growth has slowed down or ceased, a reversal of bioche changes is apparent. Five or six months after the dose of when the liver is nearly normal in size, the recovery of the o s virtually complete. 2. Biochemical Changes and Liver Enlargement in Hepatectomized Rats A. Procedure: To study the effect o liver tissue alreajT in a state of rapid growth or regenertion, six male ChR-CD rats were subjected to partial hepatectomy by removing 60 to 70# of the organ. Forty-eight houB*aftber the operation,' the rats were given a single oral dose o f M p W p f 12 mg/kg. The experiment was also conducted reversing the'br'aer of treatment by reporting 60 to 70# of the liver of rats which had been treatgd wi^h/jjBjj48 hours previously. Sham-operated animals dosed withrBB^ncThepatectomized animals -served as controls. Fourteen days after the single oral dose of 7 J the livers of the rats were removed, weighed and analyzed-Tfof* glycogen, phospholipids, DNA, RNA and alkaline phos phatase activity. B. Results: The results of these experiments are shown in Table 2. Two weeks after the operation, the livers of the partially hepatectomized control animals were normal in size for rats of this age, sex, and strain, but the alkaline phosphatase activity and DNA were higher than in iy;rm%L livers. When partially hepatectomized rats were treated with0RJ the livers grew very rapidly; two weeks after treatment wiiiT the trimer, the livers were more than twice those of the hepatectomized controls. The phos pholipid content and alkaline phosphatase activity were increased while the glycogen contentdegreased. The DNA was again decreased as in the livers of intact/jMytreated rats, but the RNA had increased. Except for this increase in RNA, the results.were in agreement with the previous measurements on livers fromjJHVltreated rats shown in Table 1. The biochemical changes in the JivST occurred whether partial hepatectomy preceded or followed treat ment w i t h U g s l although the effects were*somewhat less when hepatectomsr followed the treatment with; Company Sanitized. Does not contain TSCA C3f - 4- 3. Effect of Chemical Injury on^rfjtreated Rat Livers A, Procedure: -Six male ChR-CD rats were given a single oral dose of 12 mg, o 7 j iBKand then 48 hours later, a single sub lethal dose o equivalent to 0.1 ml per 100 gm of body IntraperltofiSally. Animals which received only served as controls. B. Results: The results of this experiment in Table 2. 6ne of the six rats dosed with1' ` died^eight days later, Jbut all six rats treat __ ^ t h e d i e d within 48 hours. No an&lysis of the Tiver was possible, ror all of the animals were found dead and post-mortem changes had begun. When examined grossly, however, the .livers were massive in size, approximately 30 to 40 gm, and yellow in color, probably from acute fatty infiltration. The rapidly enlarging liver, with a greater than normal l l p j d:glycogen ratio, is more susceptible to the toxic effect o f p W P f p h a n normal liver. Biochemical measurements on the surviving ratB showed no appreciable differences from controls two weeks after the treatment. 4 III. OP ADMINISTRATION AND IN THE RAT A. Procedure: The study was designed as an additional evaluation of the effects of combining hepatotoxic agents. The objective was to determine, by oral ^administration to rats, whether repeated sublethal doses ofy^|BBIHHflRy.would enhance the effect of a single hepatotoxic dose ofJJBPJ The outline of the study, in tabular form, is presented inPfable 3. Male ChR-CD rats, weighing between 300-400 gm, were used in the study. They were offered water and Purina Laboratory Chow on an ad libitum basis and were weighed dally. At the time of the prescribed sacrifices, the animals were subjected to grosB pathological evaluation. The liver was removed, weighed, and preserved in appropriate fixatives. B. Results: A summary of the liver weights obtained at the various scheduled autopsies lsrgiven in Table 4. The preliminary results of this study, wherein)0 P | was administered to rats in a single dose of 12 ng/kg, lmulanou8ly with, or after, their exposure tof indicate that the increase in liver weights obsdTVej experiment was not any greater than that produced by| a dose of 12 mg/kg. IV. THE EFFECT UPON PENTOBARBITAL SLEEPING TIME IN THE RAT A. Procedure: Changes in liver function or liver morphology have served as traditional indicators of hepatotoxicity. More recently, a quick and simple pharmacologic test has been developed iji I c-any Samiioezeed, Docs not contain fSGA Cw m - 5- which has been used to determine liver damage - Pentobarbital Sleeping Time (PST). * Pentobarbital is metabolized in the liver; a. change in particular liver enzymes is reflected by a prolongation, or shortening, of the sleeping time induced in animals by this com pound's normal anesthetic action. Therefore, alteration in the PST has been correlated with changes in liver function and liver morphology. When young adult male ChR-CD rats are given 30 mg kg Na pentobarbital in an aqueous solution intraperitoneally, their average sleeping time was found to be 58 + 19 minutes (for 110 rats). In studying the effect of fcWadrainistration upon PST, groups of ten rats were employed. A COnCrol value for PST was obtained for each group of ten rats, usually 24 hours before the test began. S ty-four hours later, the rats were given a single oral dose of at the rate of 12 mg/kg. PST was determined on each group of . four hours, 24 hours, 48 hours,-and at other time intervals . after the oral administration of t h e H M J Control groups were similarly examined for PST at the sarae-fime intervals. The results of two of these tests are summarized in Table 5 B. Results: The PST of rats that receivedftBljwas first prolonged; it then became shorter and shorter until, at approxi mately six days after dosing, none of the rats could be anesthetized by a dose of Na pentobarbital that still affected untreated rats. Approximately seven weeks later, the PST ofJlBiftreated rats could again be determined, but it was still much shorter than that observed in control animals. The same observations were made for the next two weeks, at the end of which time the experiment was terminated. V. EFFECT OF FLUOROCARBON SURFACTANTS ON LIVER FUNCTION IN DOGS A . Procedure: Three male-beagles from the stock colony were given a single oral dose o f[(Ml equivalent to 4, 6, or 9 mg/kg. Samples of blood were taken^&t rrequent intervals for three weeks and then weekly thereafter. The dog that received 4 mg/kg was later given doses of 26 and 60 mg/kg. Four other-yale beagles were fiven a single oral dose of e i t h e r n j ^ H M M ^ p ^ H equivalent to 30 mg/kg and a similaraeries of measurements nacre. Since both dogs receiving t h e W | B p B | died within 48 hours, the experiment was repeated with two efcneraogs which were given a 200 mg/kg dose of this-disgersing agent. The two dogs that survived the exposure to the| i H S H were given an additional oral dose, equivalent to 670 this time. The following biochemical measurements were made routinely on the blood: sugar, urea nitro sterol and alkaline >hosplaytase. When the 60 mg/ >ri^|J 470 mg/kg dose of p p nr the 670 mg/kg dose were administered, the level T a n W v i t y of lactic dehydro IDH>. isocitric dehydrogenase <ICDH), aldolase, glutamic oxalacetic (GOT) and glutamic pyruvic (GPT) transaminase were also measured. A routine hematological Company Sanitized. Does not contain TSCA CBI examination and an analysis of a 24-hour urine specimen were made at intervals on these animals. The level of the various components of the blood following the dose of fluorocarbon dispersing agents was compared with an average value observed prior to the exposure and with a similar measurement made at the same time on specimens from stock colony dogs. For the enzyme activities, a normal range was established from measurements made on a number of stock dogs. The activity was also measured at least once, prior to treatment, on the dogs dosed with the dispersing agents. B. Results: The significant biochemical and clinical findings are summarized briefly in Table 6. The principal finding-indicative of some injury or dysfunction in the dogs that receiveq_<pKfwas a decrease in the plasma chol esterol and an increase in Bbomsulfalein retention (BSP) and APase activity. The effects on cholesterol and APase are shown in Figures 2 and 3. These began to change within one week and became "abnormal", i.e., exceeded an arbitrary limit of two standard deviations flora the pre-exposure mean within three weeks after the dose was administered. The increase in BSP retention occurred during the first ten days and then began to return to normal. The depression of plasma cholesterol in all three dogs occurred earlier and began to return to normal sooner than the rise Iq ,plasma APase activity. The dog that received the highest dose of(pBB [showed the greatest change in plasma APase from the pre-exposure""!an. The activity of a number of enzymes considered to be sensitive indi cators of liver injury was measured in the plasma when a 60 mg/kg dose cfl^Blwas administered. The results of these biochemical measurements are presented in Table 7. All the plasma enzymes measured were elevated within the first three days after the dose of 60 mg/kg of l|pp[was administered. The greatest increases were in the GPT a n d w a s e . Within one week, all but these were within the normal range. The depression of the plasma cholesterol occurred more slowly, but there was no evidence of a return to normal after three weeks as occurred in the dogs receiving 6 or 9 mg/kg. With the 200 mg/kg dose of Igjttju the GPT and GOT were elevated in both dogs within 48*rours? One week later, they were in the normal range. When 450 mg/kg of this compound was admin istered, all of the enzymes measured were markedly elevated 24 to 48 hours later. The greatest change occurred in the GPT and ICDH. Both animals expired within 48 hours after dosing. The 450 mg/kg dose o f U s p U n caused elevated levels of all the enzymes measured withiiF48fiours in one (No. 2) of the two dogs exposed. On the tenth day after the dose was administered these were within the normal range. The other dog showed no effect from the treatment. At the 670 mg/kg dose of this compound, Company Sanitized. Does not contain TSCA CBj -7- Dog No. 2 showed a rise in GOT, GPT, and APase daring the first 48 hours, and a return to normal within two weeks. There was a slight rise in the APase, GPT and GOT in Dog No. 1. SUMMARY administered to rats cause - __... ... __ __ ntinues for as long as two months^after'the treatment. Morphologically, the change is characterized by an enlargement of the hepatocyte. Changes in the biochemical composition, enzyme activity and respiration, accompany this enlargement and indicate a disturbance in the normal metabolic functions of the organ. The changes in nucleic acid content and morphology of the cell suggest an interference in mitotic activity. Reversal of these changes begins in two months and is nearly complete in five to six months. Rats that have been subjected to excision of 60 to 70$ of the. `' * " or after a single oral dose of 12 mg/kg ofthan normal livers. is more toxic give _ if 12 mg/kg of from the accumulation and retention of i liver which has more fat and less glycot. that have been iis may result in the enlarging normal liver. When U J is administered to rats in a single dose of 12 mg/kg simifiraneously with or after their exposure to repeated sublethal oral doses of ethyl alcohol, the increase in IjLver^ weight observed is not any greater than that produced byjfl^) alone at the same dose. Following an initial depressant effect byfllAon enzymes that metabolize sodium pentobarbital, there is apparently an increase in the rate of metabolism of sodium pentobarbital so that it cannot exert its anesthetic action at usually anesthetic dose levels. Liver function studies in dogs have shown t h a t ^ e liver. On the least, t _______________ ________ _ ivity in of cellular damage. OnlyfiHPlflowers the cholesterol level in dogs given 4 mg or more per"Kg of body weight in a single oral dose. Of all the measurements made, alkaline phosphatase and GPT are the most sensitive in detecting an effect on the liver in dogs from all three dispersing agents. 8- - EFFECT OF FLUOROCARBON DISPERSING AGENTS ON THE LIVERS OF RATS AND DOGS Medical Research Project Nos, Report No. 123-65 HASKELL LABORATORY FOR TOXICOLOGY AND INDUSTRIAL MEDICINE Report by: \ > | u a [ ' \John R. Barnes Chifef^ Biochemistry Section Henr^ Sherman Chief, Oral Toxicity Section JRB/HS/ah 8- 19-65 Company Sanitized. Does not contain TSCA CBl TABLE 1 BIOCHEMICAL CHANGES IN RAT LIVER Body Weight gm Liver Weight gm Liver % Body Os ,ifL/hr. COa /L/hr. RQ APase BU/gm Esterase U/gm Choline Oxidase Water Ji Protein $6 Pat % Glycogen Ash Cholesterol % NPN % Phospholipids Potassium RNA % DNA % Plasma APase BU I ml Plasma Cholesterol mg# 345 13.5 3.94 7.23 5.84 0.81 2.45 245 69 68.8 18.1 4.14 4.54 1.38 0.43 0.23 3.15 0.34 0 .9 0 7 0.160 324 28.5 8.80 7.97 5.73 0.72 3.60 180 61 69.8 19.6 4.18 2.60 1.36 0.40 0.26 3.96 0.33 0.872 0 .1 2 4 546 18.0 3.30 7.00 5.67 0.81 2.66 210 65 68.0 17.2 5.03 3.86 1.36 0.44 0.21 3.29 0.42 0.834 0.176 51 93 476 27.9 5.86 6.99 5.08 0.72 2.86 245 78 68.2 18.3 5-31 2.93 1.32 0.40 0.2> 4.17 0.40 0.786 0.158 51 144 5 1/2 Months 640 18.8 2.93 6.10 4.68 0.77 3.41 74 68.3 16.7 4.87 4.44 1.36 602 21.4 3.52 5.93 4.50 0.76 3.04 78 68.3 16.9 .4.81 4.05 1.37 0.29 3.08 0.40 0.824 0.185 0.30 3.20 0.42 0.815 0.176 Company Sanitized. Does not contain T SCA CBI TABLE 2 BIOCHEMICAL CHANCE!&- ItL-RAT LIVER TISSUE 2 WEEKS TREATMENT WITH' ND AFTER TREATMENT WITH MY AND/OR Treatment 1 Treatment 2 No. Liver of Liver * Qly- Phospho- Rats Mortality Weight Body cogen APase lipids RNA DNA 0 Hepatectomy 40 12.6 5.15 4.60 5.80 5.5 0.95 0.20 Sham Operated 6 0 50.5 8.24 2.47 5.94 5.8 1.15 0.12 Hepatectomy 60 29.2 7.29 2.07 4.55 4.5 1.04 0.12 Hepatectomy 6 0 24.6 7.18 1.74 5.82 4.0 0.95 0.15 0 6 1/6 15.6 5.71 4.16 2.69 5.0 0.96 0.19 6 6/6 All animals d L within 48 hours after receiving AHT a) 12 mg # "* body weight b) 0.1 mlW 100 gm body weight Company Sanitized. Does not contain I S C A TABLE 3 DOSING AND SACRIFICE SCHEDULE OP ANIMALS GIVEN Group Control (no dosing) [2250 mg/kg/day, 5 x week for 2 weeks) -- 1(2250 mg/kg/day, jreek for 2 weeks) + ,, _l(12 m g / k g o n day of first M H M i w d o B e ) 1 ( 2 2 5 0 mg/kg/day, x week for 2 weeks) + i (12 mg/kg on day of !10th dosejj ----J ( 2 2 5 0 mg/kg/day, x week for 2 weeks) + . ^ ( 1 2 mg/kg on 14th Iday after 10th i ---- " Idose) Total No of Animals 8 8 4 Hrs After 10th Doee 2 2 Number of Animals Sacrificed________ 14 Days After 28 Days After 2 Months 10th Dose loth Dose After 10th Dose 222 222 22 2 62 2 22 Company Sanitized. Does not contain TSCA i TABLE 4 Group LIVER WJSIiGHTS OF OFa? S RECEIVING REPEAT AND A SINGLE DOSE G, >ES Liver Weights (gm) of Animals Sacrificed Hrs. A f t e r 14 Days A f t e r 26 Days After ' 10th. Dose 10th Dose 10th Dose After 10th Dose Control (no closing) 16; 15 16; 20 !9; 23 22; 16 day, 5 x week for 2 weeks) 15; -* 24j 19 25; 21 16; 22 h A ( 2250 mg/kg/day, Jp a^week for 2 weeks) + \flPU (ljg mg/kg. oh day of ^ I r a t S p H ^ dose) 25; 50 42; 32 40; 30 (2250 mg/kg/day, feek for 2 weeks) + '2 mg/fcg on day of dose) (2250 mg/kg/day, week for 2 weeks) + 1(12 mg/kg op 14th lay after 10t h 1 dose) 39; 44 36; 33 30; 41 36; 36 37; 36 1 animal died after the fourth dose Company Sanitized. Does not contain T SCA CBJ TABLE 5 Treatment Control AVERAGE SLEEPING TIMES OP RATS GIVEN Na PENTOBARBITAL INTRAPERITONEALLY AT A DOSE LEVEL OP 30 mg/kg --Pre-4E"xHpoousrusr--e--- P f SD R m sr> ft 24 "Wnurs -----P--o--s--t--irEnx--tf>toOuBrusre P S P t D ....I T PST f "SC FT 6 Days PST + 5 I8jj 49 + 14 10 52 + 8 TU 10 56 + 11 10 TU TU +` 4o1 99 T 47 + 6 10 121 + 14 10 TU TU co + VO TU 25 t 6 8 TU ? Days rat I s i 1 TT 41 + 12 sis Ho jJo--1 Control 50 + 12 ojo 47 + 8 10 47 t 6 IC 46 + 11 8 56 + 12 io TU TU (l^^^g) 56 + 15 10 143 + 18 TU 33 t 12 I 22 + 6 A 0 T = Average Pentobarbital Sleeping Time Standard Deviation (mirutes) = dumber of rats which went to sleep Total number of rat3 dosed Company Sanitized. Does not contain TSCA CM 9 Days Treatment PST + SD Control (12 mg/kg TABLE 5 (Contd) AVERAGE SLEEPING TIMES OP RATS GIVEN Na PENTOBARBITAL INTRAPERITONEALLY AT A DOSE LEVEL OP 30 mg/kg Poat-Exposure 16 Dayb 27 Days PST t SD R- PST Z SD 49 Days 56 Days 64^Pays p s t T "s d r " PST t SD R~ PST Z SD Discontinued after seven days Control 72 + 10 (l^ l^ L g ) 65 t 18 8 8 0 + 2 5 t TU 00 TU TU JTUJ 81 + 20 olo 76 + 16 10 82 t 17 10 T T 0 18 + 4 TU 7 22 b T 6 18 + 2 T T PST + SD = Average Pentobarbital Sleeping Time + Standard Deviation (minutes) R = Number cf rats which went to sleep Total number of1 rats dosed ggnipany Sanitized. Does ns? contain T SC A C Si TABLE 6 .CLINICAL AND HT.OCHEMICAL OBSERVATIONS IN DOGS ADMINISTERED FLIIOROflflRRfiN DISPERSING AGENTS No. Compound Dogs Bose 7 Rat ALD 1 1 91 6 9 4 26 60 2 200 2 450 2 450 2 670 10 15 None 7 None 0 43 None 0 100 Vomiting, anorexia, polydipsia, weakness, wt. loss, black tarry feces for 4 days_____ 30 Vomiting, polydipsia 0 0 6V Vomiting, polydipsia, 2 blood, feces and vomifcus, tremors, convulsions, anorexia,death 1-3 days 30 Vomiting, 2-4 hours 0 45 Vomiting Hypocholesterclemia, elevated APaBe Hypocholesterolemia, elevated APaBe, BSP retention Hypocholesterolemia, elevated APase, BSP retention Hypocholesterolemia, elevated APase, OPT, BSP retention Hypocholesterolemia, elevated APase, OPT, LDH, ICBH, aldo lase, jaundice, bllirubinuria Elevated APase (1/2), OOT,OPT (2/2), lowered amylase Elevated APase, GPT, ICDH, LDH, aldolase, jaundice Normal or slightly elevated cholesterol, APase, OPT, ICDH, LDH (1/2) Normal or si. elevated chol esterol, elevated GOT, OPT (2/2),si. elevated APase(1/2), amylase and sugar (2/2), ele vated after 1 week Company Sanitized. Does not contaSn TSCA CB.' TABLE T LIVER FUNCTION TESTS IN DOGS - 0* 133 2.3 35 230 8 66 5.5 26** 15 49 5.5 22 47 7.3 37 86 7.2 3 74 7 78 60 10 64 14 50 21 48 21.7 15.7 13.4 13.1 11.4 850 3900 270 145 135 91 290 77 145 * Average pre-exposure ** Previously received 4 mg/kg 6l days before a dose of 26 tng/kg 130 360 60 40 100 < 20 47 11 10 13 Company Sanitized. O^es not contain TSCA CB. TABLE 7 (Cont'd) Dose (mg/kg) 50' 50 70 Days After Dose 0* 1 2 5 7 12 . 21 27 1 2 0* 2 4 7 10 14 . 22 2 4 5 7 12 21 27 Cholesterol rcg * i2 101 112 102 93 109 111 126 126 106 112 100 127 92 109 89 106 118 164 145 112 112 126 126 102 124 128 152 100 123 121 135 142 156 142 124 159 149 144 134 124 133 102 135 127 133 140 136 APase Bod. Units 12 2.8 1.7 2.6 2.1 2.6 6.9 2.5 10.1 1.8 8.0 1.4 5.4 1.8 3.7 2.4 3.3 7.3 20.5 38.7 1.2 2.4 1.6 3.8 2.0 4.4 1.7 3.8 1.3 2.3 1.7 2.1 l.l 1.4 1.9 2.4 1.9 2.8 1.7 3.6 1.4 3.5 1.2 2.4 1.0 2.0 1.2 2.1 Average pre-exposure GPT Units i --- 42 36 38 50 56 845 108 425 56 166 40 70 36 44 42 40 280 8500 8600 25 33 30 116 33 84 27 43 40 25 66 52 70 170 58 190 42 100 34 48 36 50 36 46 GOT Units 1" 23 21 20 36 38 880 34 52 18 22 12 14 16 30 16 16 14 16 38 22 44 172 18 54 14 22 16 16 18 24 14 16 ICDH Units 1 -------- T LDH Units T ------ 5" 2100 1141300 300 3860 302 1320 153 150 72 43 116 595 74 200 145 189 68 48 not c o n ta in T S C A CBj, Company Sanitized. Does FIGURE! I I I I I I I I I I I I I I I I L CompanySanili*,.Doesno,contai,,TSCA CB( FIGURE 2 PLASMA CHOLESTEROL OF 006S RECEIVING ORAL DOSES O F j p g J ..............MEAN NORMAL RAN6E Company Sanitized, Does no! copain TSCA GB! FIGURE 3 PLASMA ALKALINE PHOSPHATASE Q f DOGS RECEIVIN6 ORAL OOSES of| J ^ J ----------- M EA N NORMAL RANCE 006 210 9m/k9 ___ I____ J ---- 6 oo 4 e- o> 006 206 Company Sanitized. Does not contain TSCA CB1 0A YS