i~AT~IONAL TOX~ICOLOGY II~OG~ AM 

Teeh~eall Report Series 

No. 46~ 

~ s£~VIc~8, o 

% ~.~'~" 

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EFFECT OF ~IIETARY I~EST~...ICT~I©H 

©H T©Z~IC©L©GY AND CA~C~N©GENES~IS 

STUDIES IN F344/H iIi:~ATI 

AHB ~C3F~ i~CE 

Uo$o ]~E~A_~11~ENT OF ~].~.akLT~ AH~ ~UI~I ~E~%riIcE~ 

F~fie ~eall~Ih 8en°~4ee 

Na~c~a~ ~m~ ~ ~a~Ih 
FOREWORD 

The National Toxicology Program (NTP) is made up of four charter agencies of the U.S. Department of 

Health and Human Services (DHHS): the National Cancer Institute (NCI), National Institutes of Health; the 

National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health; the National 

Center for Toxicological Research (NCTR), Food and Drug Administration; and the National Institute for 

Occupational Safety and Health (NIOSH), Centers for Disease Control. In July 198 I, the Carcinogenesis 

Bioassay Testing Program, NCI, was transferred to the NIEHS. The NTP coordinates the relevant 

programs, staff, and resources from these Public Health Service agencies relating to basic and applied 

research and to biological assay development and validation. 

The NTP develops, evaluates, and disseminates scientific information about potentially toxic and hazardous 

•chemicals. This knowledge is used for protecting the health of the American people and for the primary 

prevention of disease. 

The studies described in this Technical Report were.performed under the direction of the NIEHS and were 

Conducted in compliance with NTP laboratory health and safety requirements and must meet or exceed all 

applicable federal, state, and local health and safety regulations. Animal care and use were in accordance 

with the Public Health Service Policy on Humane Care and Use of Animals. The prechtonic and chronic 

studies were conducted in compliance with Food and Drug Administration (FDA) Good Laboratory Practice 

' Regulations, and all aspects of the chronic studies were subjected to retrospective quality assurance audits 

before being presented for public review. 

These studies are designed and conducted to characterize and evaluate the toxicologic potential, including 

carcinogenic activity, of selected chemicals in laboratory animals (usually two species, rats and mice). 

Chemicals selected for NTP toxicology and carcinogenesis studies are chosen primarily on the bases of 

human expo,sure, level of production, and chemical structure. The interpretive conclusions presented in this 

Technical Report are based only on the results of these NTP studies. Extrapolation of these results to other 

species and quantitative risk analyses for humans require wider analyses beyond the purview of these 

studies. Selectionper se is not an indicator of a chemical' s carcinogenic potential. 

These NTP Technical Reports are available for sale from the National Technical Information Service, 

U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA 22161 (703-487-4650). Single 

copies of this Technical Report are available without charge while supplies last from NTP Central Data 

Management, NIEHS, P.O. Box 12233, MD El-02, Research Triangle Park, NC 27709 (919-541-3419). 

Listings of all published NTP reports and ongoing studies are also available from NTP Central Data 

Management. The Abstracts and other study information for 2-year studies are also available at the NTP's 

World Wide Web site: http//ntp-server.niehs.nih.gov. 
,~ ~--'-" ....... ~.-~ .......... ~ ......... .~ ~---~-~ ::-~ ........ ~ ........... ~ ~ .... "-~==~--~ -L`~L~-~-'='/ " ::_~: 

NTIP TECHN]ICAL ~E~O~T " 

~N THE 

. 

EFFECT ©]~ ~]IETAXY ~E~TX]ICT]I©N 

ON T©i~]IC©L©GY ANI~ CAXC[N©GENES]IS 

STUI~IE~ iN F$44/N ~AT~ i 

AND Z~C~F~ M]ICE [ 

f 

NAT]~ONAL TOX]~COLOGY ]?]~OG]~LA]V~ 

]~o~. ~v~ ~.~33 ~ 

]Research Tr~anglle ]?ark~ NC Z77%9 

i 

i 

[ 

Sep~:e~berr ]1997 

NT~ T~ 4~% 

NIH Publication No. 97-3376 

Uo$. DE]PA~T~ENT OF HEALTH AND HU1V~AN ~E]~V]~CES 

]~ub~c Hea~h ~erv~ce 

Na~na~ ]~nsfi~u~es ~f Hea~h 
2 Dietary Restriction, NTP TR 460 

CONTRIBUTORS 

National Toxicology Program 

Evaluated and interpreted results and reported findings 

K.M. Abdo, Ph.D., Study Scientist 

F.W. Kari, Ph.D., Study Scientist 

D.A. Bridge, B.S. 

J.R. Bucher, Ph.D. 

L.T. Burka, Ph.D. 

R.E. Chapin, Ph.D. 

M.Ro Elwell, D.V.M., Ph.D. 

J.R. Hailey, D.V.M. ' 

J.K. Haseman, Ph.D. 

R.A. Herbert,.D.V.M., Ph.D. 

G.N. Rao, D.V.M., Ph.D. 

J.H. Roycroft, Ph.D. 

G.S. Travlos; D.V.M. 

D.B. Waiters, Ph.D. 

K.L. Witt, M.S., Oak Ridge Associated Universities 

Southern Research Institute 

Conducted butyl benzol phthalate and t-butylhydroquinone studie~, 

evaluated pathology findings 

J.D. Prejean, Ph.D., Principal Investigator 

D.G. Scrota, Ph.D., Principal Investigator 

H.D. Giles, D.V.M., Ph.D, 

C. Lindamood III, Ph.D. 

Conducted salicylazosulfapyridine studies, evaluated pathology 

findings 

LD. Prejean, Ph.D., Principal Investigator 

H.D. Giles, D.V.M., Ph.D. 

C. Lindamood III, Ph.D. 

Battelle Columbus Laboratories • 

Conducted scopolamine hydrobromide trihydrate study, evaluated 

pathology findings 

P.J. Kurtz, Ph.D., Principal Investigator 

A,C. Peters, D.V.M., Principal Investigator 

J.D. Johnson, Ph,D. 

A.W. Singer, D.V.M. 

Experimental Pathology Laboratories, Inc. 

Provided pathology quality assurance 

J.F. Hardisty, D.V.M., Principal Investigator 

S. Botts, D.V.M., Ph.D. 

E.T. Gaillard, D.V.M., M.S. 

M. Wells, D.V.M. 

NTP Pathology Working Group 

Evaluated slides, prepared pathology report on butyl benzyl 

phthalate (I0 January 1995) 

P.K. Hildebrandt, D.V.M., Chairperson ~ 

PATHCO 

M.R. Elwell, D.V..M., Ph.D. 

National Toxicology Program 

E.T. Gaillard, D.V.M., M.S. 

Experimental Pathology Laboratories, Inc. 

J.R. Hailey, D.V.M. 

National Toxicology Program 

R.A. Herbert, D.V.M., Ph,D. 

National Toxicology Program 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

C.C. Shackelford, D.V.M., M.S., Ph.D. 

Experimental Pathology Laboratories, Inc. 

B. Short, D.V.M., Ph.D. 

SmithKline Beecham 

Evaluated slides, prepared pathology report on t-butylhydroquinone 

(9 January 1995) 

J.C. Seely, D.V.M., Chairperson 

PATHCO 

S. Botts, D.V.M., Ph.D. 

Experimental Pathology Laboratories, Inc. "' 

M.R. Elwell, D.V.M., Ph.D. 

National Toxicology Program 

J.R. Hailey, D.V.M. 

National Toxicology Program 

R.A. Herbert, D.V.M., Ph.D.. 

National Toxicology Program 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

B. Short, D.V.M., Ph.D. 

SmithKline Beecham 
Dietary Restriction, NTP TR 460 

Evaluated slides, prepared pathology report on 

salicylazosulfapyridine for rats (9 February 1995) 

M.P. Jokinen, D.V.M., Chairperson 

Pathology Associates, Inc. 

S. Botts, D.V.M., Ph.D. 

Experimental Pathology Laboratories, Inc. 

M.R. Elwell, D.V.M., Ph.D. 

National Toxicology Program 

E.T. Gaillard, D.V.M., M.S. 

Experimental Pathology Laboratories, Inc. 

J.R. Hailey, D.V.M. 

National Toxicology Program 

• R.A. Herbert, D.V.M., Ph.D. 

National Toxicology Program 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

B. Short, D.V.M., Ph.D. 

SmithKline Beecham 

Evaluated slides, prepared pathology report on 

salicylazosulfapyridine for mice (17November 1994) 

M.P. Jokinen, D.V.M., Chairperson 

Pathology Associates, Inc. 

D. Dixon, D.V.M., Ph.D. 

National Toxicology Program 

J.R. Hailey, D.V.M. 

National Toxicology Program 

R.A. Herbert, D.V.M., Ph.D. 

National Toxicology Program 

J.R. Leininger, D.V.M., Ph.D. 

Chemical Industry Institute of Technology 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

M. Wells, D.V.M. 

Experimental Pathology Laboratories, Inc. 

~,.~ ......... :~ ~ ~ ~ ~-~ ~ ~ ~ : -~ -. ~ 

Evaluated slides, prepared pathology report on scopolamine 

hydrobromide trihydrate (8 December 1994) 

M.P. Jokinen, D.V.M., Chairperson 

Pathology Associates, Inc. 

S. Botts, D.V.M., Ph.D. 

Experimental Pathology Laboratories, Inc. 

R. Cattley, V.M.D., Ph.D. 

Chemical Industry Institute of Ttchnology 

M.R. Elwell, D.V.M., Ph.D. 

National Toxicology Program 

R.A. Herbert, D.V.M., Ph.D. 

National Toxicology Program 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

Dyn~mac Corporatior~ 

Prepared quality assurance audits 

S. Brecher, Ph.D., Principal Investigator 

Ana~ytic~ Sciences, ~nc. 

Provided statistical analyses 

R.W. Morris, M.S., Principal Investigator 

S.R. Lloyd, M.S. 

N.G. Mintz, B.S. 

~otechn~cz~ Services~ ]Inc. 

Prepared Technical Report 

S.R. Gunnels, M.A., Principal Investigator 

L.M. Harper, B.S. 

E.S. Rathman, M.S. " 

D.C. Serbus, Ph.D. 

W.D. Sharp, B.A., B.S. 

S.M. Swift, B.S. 
4 

CONTENTS 

,~J~S'I'R~CT..... ~... ~................ , .: s 

• 

. 

TECHNICAL REPORTS REVIEW SUBCOMMrITEE :. ...... ~ ......... ,.. ~ ..... . .: .. 12 

~ARY OF TECHNICAL REPORTS REVIEW SUBCOMMITrEE .COMMENTS .......... 13 

INTRODUCTION.....~ .... ~... i.. ......... ~ ........ ..... ~. ......... ....... 17 

" ~ ~ 

MATERIALS AND METHODS- 21 . 

. 

RESULTS .~.~....i..... ~'''I'I':'''''~)'''''''" .... ...i, ..... . ...... .~. .... , 33 

DISCUSSION AND CONCLUSIONS ~.., .,.... .......... ; ...... ;. '.. ..... ~... ~ .. ...... . 89 , 

• . 

.... .. ..... : , i .......... ...... :..., : ..... • ................ 97 

• . 

APPENInX A • Summary of Lesions in Male Rats in the Dietary Restriction Study . • " 

• :ofButylBenzylPhthalate ....~ ..... i" ~;'.)':'~''. ...... .,.; ....... . 101 

• . 

APPENDIX B Summary of Lesions in Female Rats in the Dietary Restriction Study 

• 

of Butyl Benzyl Phthalate .......................... " . . . 137 

APPENDIX C Summary of Lesions in Male Rats in the Dietary Restriction Study 

of t-Butyihydroquinone " ,. " 171 

APPENDIX D Summary of Lesions in Female Rats in .the Dietary Restriction Study 

of t-Butylhydroquinone ....... "....... ................ •............. 207 

A~i'ENDIX E Summary of Lesions in Male Ratsin the Dietm-y Restriction Study 

of Salicylazosuifapyridine • ' • 239 • 

• 

APPENDIX F Summary of-Lesions in Male Mice in the DietarY Restriction Study 

of Salicylazosulfapyridine . .......... ' ...... ,.. ..... "............... :. 281 : 

• 

• . 

• . 

APPENDIX G : Summary Of Lesions in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate ..... - ......................... 311 

APPENDIX H Summary of Lesions in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate.. .................... • ........ 339 . . • . . 

• . 

APPENDIX I Organ Weights and Organ-Weight-to-Body:Weight Ratios .................. 369 • , . 

~ • • 

APPENDIX J Mean Body Weight and Survival Results ' .~. ~ . . . 381 

- - . ~ ~ . 

APPENDIX K Feed and Compound Consumption in the Dietary Restriction Studies ........... 399 

• • 
AAST 

Studies were conducted to compare outcomes when 

four chemicals were evaluated under typical NTP 

bioassay conditions as well as under protocols 

employing dietary restriction. Specific experiments 

were designed to evaluate the effect of diet restriction 

on the sensitivity of the bioassay toward chemicalinduced chronic toxicity and carcinogenicity and to 

evaluate the effect of weight-matched control groups 

on the sensitivity of the bioassays. Two chemicals, 

butyl benzyl phthalate and t-butylhydroquinone, were 

administered in feed; one chemical, salicylazosulfapyridine, was administered in corn oil by gavage; and 

one chemical, scopolamine hydrobromide trihydrate, 

was administered in distilled water by gavage. In 

each of four protocols, the effects of the chemical 

were assessed by a comparison between a group 

exposed to a single dose concentration of the study 

chemical and a nonexposed control group. F344/N 

rats and B6C3F 1 mice were fed NIH-07 diet eider 

ad libitum or in amounts that restricted mean body 

weights according to the following design requirements. For the core bioassay, groups of 50 to 

60 ad libitum-fed animals were allotted to a control 

group and three dosed groups for approximately 

104 weeks or up to 128 weeks (t-butylhydroquinone 

study). The comparison between the control group 

and the group receiving the highestdose was used to 

represent the outcome of the bioassay under 

ad libitum feeding protocols. In a second comparison, 

outcomes from the group receiving the highest dose 

were compared with a weight-matched group of 50 to 

60 untreated controls; the weight-matched controls 

received feed in amounts restricted so that the mean 

body weight matched the mean body weight of the 

dosed group. , 

Two additional groups of 48 to 60 animals (one 

control and one dosed group) were offer.ed feed in 

amounts that limited the mean body weight of the 

control group to approximately 85% that of the 

controls fed ad libitum under the first protocol. 

Animals assigned to this dietary restriction paradigm 

were evaluated after 104 weeks or 130 weeks 

(t-butylhydroquinone). A fourth protocol was em- • 

• . 

ACT 

loyed to evaluate whether an additional period of 

exposure (up to 1 year) would influence the neoplasm 

profile of animals fed a restricted diet. Tw0groups of 

approximately 50 animals (one control and one dosed 

group) in the butyl benzyl phthalate, salicylazosulfapyridine, and scopolamine hydrobromide trihydrate 

studies received restricted diets, as under the third 

protocol, for 3 years or until survival in either group 

was reduced to 20 %. 

Butyl benzyl phthalate caused an increased incidence 

of.pancreatic acinar cell neoplasms in ad libitum-fed 

male rats relative to ad libitum-fed and weight-matched controls. This change did not occur in rats in 

the restricted feed protocol after 2 years; however, 

acinar.cell adenomas were observed in three exposed, 

feed-restricted males at 30 months. Feed restriction 

is known to influence the incidence of pancreatic 

acinar cell neoplasms and may have prevented the full 

expression of this chemical-induced effect. Butyl 

benzyl phthalate also caused an increased incidence of 

urinary bladder neoplasms in female rats in the 

32-month restricted feed protocol. The incidences of 

urinary bladder neoplasms were not significantly 

increased in female rats in any of the 2-year protocols, 

suggesting that the length of study, and not body 

weight, was the primary factor in the detection of this 

carcinogenic response. 

Salicylazosulfapyridine caused an increased incidence 

of urinary bladder papillomas in male rats fed 

ad libitum relative to ad libitum-fed and weightmatched controls. This increasewas associated with 

an increased incidence of urinary bladder calculi; the 

incidences of urinary bladder concretions, dilatation, 

and hyperplasia were also increased in dosed males. 

The incidences of urinary bladder papillomas and 

calculi were not increased in male rats. receiving 

salicylazosulfapyridine that were fed restricted diets. 

In male mice, .salicylazosulfapyridine caused an 

increased incidence of liver neoplasms relative to the 

,ad libitum-fed and weight-matched controls. This 

increase did not occur in the restricted feed protocols. 

 ̧', 
6 

Liver neoplasms in mice are greatly influenced by 

body :weight, and the marked mean body weight 

reduction observed in dosed male mice in the 

restricted feed protocols may have overridden the 

carcinogenic response. 

Neither t-butylhydroquinone nor scopolamine hydrobromide trihydrate caused increased neoplasm incidences under any of the experimental protocols. 

Results consistently show that feed restriction caused 

decreased incidences of neoplasms and nonneoplastic 

lesions at a variety of anatomic sites in control and 

dosed animals. Furthermore, the sensitivity of the 

bioassay to detect a carcinogenic response was altered 

by dietary restriction: two of the four chemicals 

caused increased incidences of neoplasms at three 

sites when evaluated under a standard ad libitum 

fegding protocol for 104 weeks. When control and 

:-dosed groups were subjected to dietary restriction, ~, 

none of these three sites was detected as a target of 

carcinogenesis after 2 to 3 years. Rather, one different site of carcinogenesis was detected after 

32 months. When dosed animals in the ad libitum 

feeding protocol were compared to weight-matched 

control groups, three sites were identified as targets of 

carcinogenesis and corresponded to the three sites 

discovered under the ad libitum feeding protocol. . ~ 

DietaryRestriction, NTP TR 460 

The magnitude of the response was greater when the 

weight-matched controls protocol was used. Dietary 

restriction of dosed and control animals decreased the. 

sensitivity of these carcinogenesis bioassays. 

Regarding the future use of dietary restriction regimens in long-term studies, only limited conclusions 

can be drawn because only four chemicals were 

evaluated and none of these proved to be a strong 

carcinogen. However, the results of these studies are 

consistent with previous findings that dietary restriction increases survival rates and decreases the incidences of neoplasms and nonneoplastic lesions at a 

variety of sites in rats and mice. This association 

between reduced body weights and decreased neoplasm incidences underlines the necessity that the 

doses selected for chronic studies not exceed "minimally toxic doses" so that no marked body weight 

reductions (or increases) will occur in the dosed 

groups. Such body weight changes complicate the 

detection of carcinogenic effects. 

The following tables summarize and compare the 

findings from ad libitum-fed, weight-matched, and 

feed-restricted groups for each chemical. 

A summary bf the Technical Reports Review Subcommittee comments and the public discussion on this Technical Report appears on page 13. 
~[~'y ~c~o~,~T]~ T~L ,~0 7 

~m~-y o~ ~he Die~u'y lI~es~ric~tion S~u~y of Bu~ty~ ~e~y~ ~h~h~e~e 

Ad Libitum 

Feeding 

MAL~ ~AT$ 

Doses 0 or 12,000 ppm in 

feed 

~ody weights c 417 g, 379 g 

~urv~v~ r~£es 28•50, 22•50 

~onneopl~sfic . Pancreas facinus): 

effec~ 

I~eop~stic effec~ 

FE~.,E ]RATS 

Doses 

~y we|gh~s 

Survived re~tes 

~onn~op~stic 

~ff~cts 

I~leopl~st|c effecf.s 

hypcrplasia (4•50, 

12/50) 

Pancreas ~acinus): 

.adenoma (3/50, 10/50) 

0 or 24,000 ppm in 

feed 

225 g, 199 g 

25•50, 29•50 

Urinary bladder: 

transitional epithelium, 

hyperplasia (4/50, 

10/50) 

None 

Weight-Matche¢~ 

Controls a 

0 or 12,000 ppm in 

feed 

377 g, 379 g 

34•50, 22•50 

~ncreas (acinus~: 

hyperplasia (2•50, 

12/50) 

Pancreas ~acinus): 

adenoma (0/50, 10/50) 

0 or 24,000 ppm in 

feed 

203 g, 199 g 

41/50, 29•50 

Urinary bladder: 

transitional epithelium, 

hyperplasia (0/50, 

10/50) 

None 

Restricted Feed 

(2 Years) 

0 or i2,000 ppm in 

feed 

355 g, 336 g 

34•50, 31150 

None 

• None 

0 or 24,000 ppm in 

feed 

187 g, 175 g 

35150, 39150 

Urinary bladder: 

transitional epithelium, 

hyperplasia (0•50, 

14/50) 

None 

a Includes exposed group from ad libitura feeding protocol 

b Survival fell to 20% at 30 months (males) or 32 months (females) 

c Body weight data are presented as the average of weekly mean body weights for weeks 14 through 52: 

Restricted Feed 

(Lifetime b) 

0 or 12,000 ppm in 

feed 

363 g, 340 g 

10/50, 13/50 

None 

None 

0 or 24,000 ppm in 

feed 

189 g, 175 g 

I0150~ 11150 

Urinary bladder: 

~ transitional epithelium, 

hyperplasia (0/49, 

16/50) 

Urinary bladder: 

papilloma'or 

carcinoma (1/49, 6/50) 
8 Dietary Restriction, NTP TR 460 

Summary of the Dietary Restriction Study of t-Butylhydroquinone • 

Ad Libitum Weight-Matched 

Feeding. " Controls a 

MALE RATS 

Doses " 0 or 5,000 ppm in feed 0 or 5,000 ppm in feed" 

Bod3~~weights b 425 g, 390 g 378 g, 390 g 

Survival rates 8/60, 14/60 12/60, 14/60 

Nonneoplastic effects None None 

Neoplastic effects None None 

FEMALE RATS 

Doses 0 or 5,000 ppm in feed 0 or 5,000 ppm in feed 

Body weights 232 g, 211 g 213 g, 211 g 

Survival rates 10160, 17160 22/60, 17/60 

Nonneoplastic effects None • None 

Neoplastic effects None None 

a Includes exposed group from ad libitumfeeding protocol' 

b Body weight data are presented as the average of weekly mean body Weights for weeks 14 through 52. • 

, . 

Restricted Feed 

(30 Months) 

0 or 5,000 ppm in feed 

365 g, 361 g 

10/60, 22/60 

None 

None 

0 or 5,000 ppm in feed 

196 g, 196 g 

18160, 24160 • • 

None 

None 
. . . . . . . ~.... . 

~, Dietary Restriction, NTP TR 460 

Summary of the Dietary Restriction Studies of SaHcylazosulfapyr~d~ne 

MALE RATS 

Doses 

Body weights b 

Survival rates 

Nonneoplastic 

effects 

Neoplastic effects 

Ad Libitum 

Feeding 

0 or 337.5 mg/kg in 

corn oil by gavage 

410 g, 399 g 

35/50, 23/50 

Urinary bladder: 

calculus (0/50, 27/50); 

concretion (0/50, 

10/50); dilatation 

(0/50, 7/50); mucosa, 

hyperplasia (0/50, 

41/50) 

Kidney: concretion 

(0/50, 33/50); 

hydronephrosis (0~50, 

28/50); mineralization 

(3/50, 13/50); renal 

tubule dilatation (0/50, 

11/50); transitional 

epithelium; hyperplasia 

(10/50, 43/50) 

Spleen: hematopoietic 

cell proliferation 

(14/50, 23/50); 

hemosiderin 

pigmentation (14/50, 

30/50) 

Urinary bladder: 

papilloma (0/50, 6/50) 

Weight-Matched 

Controls a 

0 or 337.5 mg/kg in 

corn oil by gavage 

408 g,'399 g 

31/50, 23/50 

Urinary bladder: 

calculus (0/50, 27/50); 

concretion (0/50, 

10/50); dilatation 

(1/50, 7/50); mucosa, 

byperplasia (0/50, 

41/50) 

Kidney: concretion 

(0/50, 33/50); 

hydronephrosis (0/50, 

28/50); mineralization 

(6/50, 13/50); renal 

tubule dilatation (1/50, 

11/50); transitional 

epithelium, hyperplasia 

(5/50, 43/50) 

Spleen: hematopoietic 

cell proliferation 

(9/50, 23/50); 

hemosiderin 

pigmentation (20/50, 

30/50) 

Urinary bladder: 

papilloma (0/50, 6/50) 

Restricted Feed 

(2 Years) 

0 or 337.5 mg/kg in 

corn oil by gavage 

346 g, 330 g 

34/51, 39/50 

Urinary bladder: 

transitional epithelium, 

hyperplasia (0/51, 

7/50) 

Kidney: concretion 

(0/51, 22/50); 

mineralization (2/51, 

11/50); transitional 

epithelium, hyperplasia 

(3/51, 18/50) 

Spleen: hemosiderin 

pigmentation (12/51, 

35/50) 

None 

Restricted Feed 

(30 Months) 

0 or 337.5 mg/kg in 

corn oil by gavage 

348 g, 329 g 

10/49, 24/50 

Urinary bladder: 

transitional epithelium, 

hyperplasia (0/49, 

8/49) 

Kidoey: concretion 

(0/49, 35/50); 

nephropathy (39/49, 

48/50); trransitional 

epithelium, hyperplasia 

(1/49, 37/50) 

Spleen: hemosiderin 

pigmentation (15/49, 

33/49) 

None 

I 

I: 

I 

![ 

i 
10 Dietary Restriction, NTP TR460 

Summary of the Dietary Restriction Studies of Salicylazosulfapyridine (continued) , 

Ad Libitum 

Feeding 

Weight-Matched 

Controls a 

Restricted Feed 

(2 Years) 

Restricted Feed 

(3 Years) 

MALE MICE 

Doses 0 or 2,700 mg/kg in 

corn oil by garage 

0 or 2,700 mg/kg in 

corn oil by garage 

0 or 2,700 mg/kg in 

corn oil by garage ~ 

0 or 2,700 mg/kg in 

corn oil by gavage 

Body weights 45.0 g, 38.3 g 39.4 g, 38.3 g 39.2 g, 32.0 g 38.4 g, 32.2 g 

Survival rates 40/50, 46/50 45•50, 46/50 42/52, 44/50 20148, 34•50 

Nonneoplastic 

effects 

Liver: clear cell focus 

(2/50, 11/50); 

eosinophilic focus 

(6/50, 22/50) 

Liver: clear cell focus 

(2/50, 11150); 

eosinophilie focus 

(1/50, 22/50) 

None None 

Neoplastic effects Livgr: hepatocellular 

adenoma (13150, 

42/50) 

Liver: hepatoeellular 

adenoma (8/50, 42/50) 

None None 

a Includes dosed group from ad libitum feeding protocol 

b Body weight data are presented as the average of weekly mean body weights for weeks 14 through 52. 
11 

V 

. . . . . 

Die~ry Restriction, NTI? TR ,~60 

Sun~m~ry of the Dietary ]Restriction Study of Scopo~ni~e ~ydrobrom~de Tr~hy~rate 

Ad Libitum Weight-I~atched Restricted Feed Restricted Feed 

Feeding Controls a (2 Years) (3 Years) 

MALE MICE 

Doses 0 or 25 mg/kg in water 0 or 25 mg/kg in water 0 or 25 mg/kg in water 0 or 25 mg/kg in water 

by garage by garage by garage by gavage 

Body weights b 45.0 g, 36.0 g 35.9 g, 36.0 g 31.3 g, 29.1 g 31.9 g, 29.2 g 

Survival rates 40/50, 39/50 41/50, 39/50 49/50, 48/50 28/50, 37/50 

Nonneoplastic None None None None 

effects 

Neoplastic effects None None None None 

~MALE MICE 

Doses 0 or 25 mg/kg in water 0 or 25 mg/kg in water 0 or 25 mg/kg in water 0 or 25 mg/kg in water 

by garage by garage by garage by gavage 

Body weights 43.2 g, 34.8 g 32.3 g, 34.8 g 29.2 g, 27.8 g 29.9 g, 27.2 g 

Survival rates 33/51, 38/51 36/50, 38/51 47/50, 44/50 20/50, 19/50 

Nonneoplastic None None None None 

effects 

Neoplastic effects None None None None 

a Includes dosed group from ad libitum feeding protocol 

b Body weight data are presented as the average of weekly mean body weights for weeks 14 through 52. 
12 Dietary Restricti0n~ NTP TR460 

.. 

NATIONAL TOXICOLOGY. PROGRAM BOARD OF SCIENTIFICCOUNSELORS ~ 

TECHNICAL REPORTS REVIEW SUBCOMMITrEE ' , 

. . 

• • . 

• 

~ , 

The members of the Technica! Reports Revie~y Subcommittee who e~'aluated the draft NTP Technical Report O n the effect ofdietary. ~ 

res2ietion on toxicity and carci,nog9nesis studies in F344/N rats and B6C3Ft mice on 20 June 1995 are listed below. Subcommittee members 

serve as independent scientists, not as representatives of any institution, company, or governmental agency. In this capacity, Subcommittee 

members ha~,e.five major responsibilities in reviewing NTP studies: " .. • 

• • . • 

• to ascertain that all relevant literature data~have been adequately cite~l and interpreted, " .. . .. 

• to determine if the desigh and conditions of the NTP studies were appropriate, 

• to ensure that the Technical RePort presents the experimental results and conclusions fully and clearly,. 

. ¶ tO judge the significance of the experimental results by scientific criteria, and ..... . 

• to assess the evaluation of the evidence of carcinogenic activity and other observed ~oxic: responses. 

. 

• 

Arnold L. Brown, M.D., Chairperson 

University of Wisconsin Medical School 

Madison, WI 

Thomas L. Goldsworthy, Ph.D. ,~ Principal Reviewer 

Department of Experimental Pathology and Toxicology 

• Chemical Industry Institute of Toxicology 

Research Triangle Park, NC . : 

Ronald W. Hart, Ph.D., Special Reviewer ' 

National Center for Toxicological Research ~? ,. 

: Jefferson, AR . • . . . . " ...; 

MerylH. Karol, Ph.D. 

Deparlment of Environmental Occupatibnal H~lth 

. University of Pittsburgh ' . 

Pittsburgh, PA • : 

Curtis D. Klaassen, Ph.D. 

Department of Pharmacology and Toxicology • ~ " " 

University of Kansas Medical Center 

Kansas City, KS 

• 

Claudia S. Miller, M.D., M.S. 

University of Texas Health Sciences Center " 

'San Antonio, TX :" 

, '. 

Janardan K. Reddy, M.D. 

Department of Pathology 

Northwestern University Medical School 

Chicago, IL 

Irma Russo, M.D. 

Fox Chase Cancer Center. . ... 

Philadelphia, PA " • . 

.. 

Louise Ryan, Ph.D. 

Division of Biostatistics 

Harvard School of Public Health and 

Da~-Farber Cancer Institute 

Boston, MA 

Robert E. Taylor; M.D., Ph.D. 

Deparunent of Pharmacology 

Howard University College of Medicine 

Washington, DC 

Mary Jo Vodicnik, Ph.D. 

Lilly MSG Development Center 

Belgium 

Jerrold M. Ward, D.V.M., Ph.D. 

National Cancer Institute 

Frederick, MD' 

• ~. 

Richard Weindruch, Ph.D., Special Reviewer 

Department of Medicine, Institute on .Aging 

University of Wisconsin 

Madison, WI 

 ̧': 

./ 

.. 

.. 
Dietary Restriction; NTP TR 460 13

. 

SUI~MARY OF TECHNICAL IILEI~O~TS. ~?dEVI~EW SUBCOMMITTEE C~IV~IV~ENTS 

On 20 June 1995, the draft Technical Report on the 

effect of dietary restriction on toxicology .and carcinogenesis studies in F344/N rats and .B6C3F~ mice 

received public review by the National Toxicology 

Program's Board of Scientific Counselors' Technical 

Reports Review Subcommittee. The review .meeting 

was held at the National Institute of Environmental 

Health Sciences, Research Triangle Park, NC. 

Dr. F.W. Kari, NIEHS, introduced the studies by 

noting that it has been recognized since the turn of the 

century that body weight reductions or feed restriction 

with concomitant decreases in body weight results in , 

increased longevity and decreased incidences of a 

variety of neoplasms and nonne0plastic lesions. He 

showed an abbreviated list of literature reports, indicating that this phenomenon is not unique to a particular species, strain, sex, tumor site, or carcinogen. 

Dr. Kari then reviewed, the primary objectives and 

overall design of the studies with the four chemicals, 

butyl benzyl phthalate, t-butylhydroquinone, salicylazosulfapyridine, and scopolamine hydrobromide 

trihydrate, and described the four basic comparisons 

made in the studies. Dr. Kari provided an overview 

of the results, discussing discordance or disparities 

between the outcomes under the various protocols. 

Dr. Kari then discussed .neoplasm sites partictilarly 

with regard to biological plausibility of weight reduction versus chemical exposure as determinants of 

incidence for certain neoplasms. 

Dr. Goldsworthy, ihe principal reviewer, stated that 

the experimental results were predictable, given the 

preexisting literature and especially the limited 

responses seen with the four chemicals. The study 

corroborates earlier evidence that increased survival 

and decreased incidences of certain neoplasms occur 

in studies With dietary restriction. His major criticism 

was that the selected chemicals limited the number of 

insights and conclusions that could be made. He said 

that both weak and strong carcinogens should have 

been used and that the chosen chemicals should have 

targeted tissues that are sensitive to dietary restric- , 

tion and that have low spontaneous neoplasm incidences, where changes due to dietary restriction in 

untreated animals normally could not be. observed. 

He said this was not the case in the current studies, 

,and it was not clear how the chemicals were chosen. 

Dr~ Goldsworthy pointed out that the studies were 

properly conducted, and he thought that Some of the 

interesting insights of the studies were obtained by 

examining the limited responses or subtle differences 

that were detected. These insights were important 

because there is a need both ,in the literature and in 

future studies for determining the effects of dietary 

. restriction on very small and variable changes after 

long-term chemical administration. 

Dr. Weindruch, a special reviewer, prefaced his 

comments by saying they should be viewed as those of 

a gerontologist with a long-term interest in the 

retardation of aging and diseases by dietary ~ restriction. His main scientific concern involved the lack Of 

a precise definition of the ad libitum feed intake and 

that the methods described did not lend confidence 

that the intake was precisely measured. In his 

experience With many. strains of rats and mice, 

ad libitum feed intake varied considerably between 

animals. Thus, with a target of 15% mean body 

weight, reduction, there would be a large range of 

individual intake values, and the use of group housing 

added to this problem. Dr. Weindruch spoke against 

the stated implication that dietary restriction "works" 

by preventing obesity, and he spoke for diets enriched 

in vitamins, minerals, and amino acids, so as to 

balance the intakes of dietary essentials among rodents 

fed different levels of calories and undergoing toxicology testing. Finally, he said that the scientific rationale for the choices of the test chemicals, doses, and 

routes of administration needs to be stated ~clearly in 

this Technical Report. 

Dr. Hart, the second special reviewer, .said his foremost criticism had to do with the choice of test 

• chemicals, commenting that if he were going to test a 

new paradigm for conducting bioassays, he would not 

randomly choose four chemicals for evaluation. 

Dr. Hart commented that the use of a maximum 

tolerated dose determined in ad libitumofed animals to 

calculate doses in animals fed restricted diets is 

misleading at best, as toxic endpoints can be more 

severely impacted by feed restriction than by 

carc~nogenicity. He said that using weight-restricted 

controls fails to take into consideration the impact that 

i 

I

J 

:I 

I 

i 

! 

I ~ 

:! 
14 Dietary Restriction, NTP TR 460 

altered caloric intake can have on a number of key 

physiological, metabolic, biochemical, and molecular 

parameters, e.g., polydipsia, increased renal clearance, or alteration of key drug metabolizing enzymes 

in feed-restricted animals. Dr. Hart found disconcerting a perceived lack of concern by the investigators that the data, in his view, fly in the face of 

50 years of similar studies, conducted in over 20 laboratories, using over 30 different model carcinogens, 

which have shown that in general, dietary restriction 

delays the onset or reduces the severity of neoplastic 

changes but does not completely eliminate such 

changes. He felt it is also important to note that 

where chemically induced neoplasms appeared to be 

eliminated in the current studies, the mean body 

weights of the dosed groups were significantly less 

than those of the corresponding feed-restricted control 

groups. ~This compromised the assumption that the 

neoplasms had really been eliminated. Dr. Hart stated 

that his main point and, as he viewed it, the main 

point of this Technical Report is that if dietary restriction is used, it should be moderate. Furthermore, to 

enhance interstudy reproducibility, a more physiological normalizer such as adjusting dietary intake to 

achieve an idealized body weight curve will be 

needed. He proposed that a small workshop be 

convened to discuss and decide what an idealized 

body weight curve is, how to achieve it, and how to 

monitor it. The findings and recommendations could 

be reported back to the NTP Board. 

Dr. Kari acknowledged the suggestions concerned 

with using more idealized conditions. However, he 

stated that the primary purpose of the dietary restriction studies was to create a data base that would help 

to clarify results retrospectively when there were 

alterations in body weight (presumably due to primary 

or secondary chemical effects) and to guide the 

interpretation of prospective studies in which alterations in body weight are expected. Thus, the experimental conditions in these dietary restriction studies 

needed to mimic those used in the bioassays, such as 

group housing and standardized diet. He said there is 

a definite lack of consensus in the literature as to the 

best experimental conditions, and it is important to 

have a data base that allows interpretations of effects 

that are often subtle. With regard to the chemicals 

selected for study, Dr. Kari said selection was based 

in part on neoplasms and nonneoplastic lesions 

expected to be induced by the particular chemicals 

selected, based on the information available at the 

time of selection. The chemicals in this set of studies 

were representative of the majority of chemicals tested 

by NTP; indeed, potent multisite carcinogens are 

exceptional. 

Dr. Hart noted that the fact that the dietary restriction 

paradigm works under diverse conditions suggests that 

body weight is a factor to be considered in making an 

evaluation of toxicity. He thought the NTP study 

could serve as a good baseline; however, better model 

compounds are needed to test the paradigm. 

Dr. Weindruch said the driving force is the caloric 

intakeper se. Dr. Karol stated that it is important to 

look at the mechanisms of effects seen in dietary 

restriction studies. 

Dr. K. Keenan, Merck Research Laboratories, said 

his laboratory is already using dietary restriction; 

however, it is called "proper nutrition" in studies with 

Sprague-Dawley rats. He said the percent restriction 

is irrelevant, but what is important is the number of 

kilocalories consumed per rat per day. He showed 

data from studies in his laboratory and the Wistar 

Institute correlating kilocalories per day the with the 

percentages of animals bearing neoplasms and neoplasms per rat. He stated that ad libitum feeding is 

one of the most adverse events to which an animal can 

be subjected. Dr. Keenan conchideed by summarizing 

the positive effects (and the lack of adverse effects) of 

moderate dietary restriction on animal health, longevity, and spontaneous and chemical-induced neoplasm incidences at his laboratory. 

Dr~ Miller stated that she supported bringing together 

experts in nutrition, geriatrics, and toxicology to focus 

on the issues around dietary restriction and toxicology 

studies. Dr. G.W. Lucier, NIEHS, agreed it would 

be a good idea for the NTP to sponsor a workshop to 

address these issues. The findings and recommendations could be commented on in an open meeting, 

perhaps through the NTP Board. Chemical selection 

would be an important issue. Dr. J.R. Bucher, 

NIEHS, commented on the increasing body weights of 

F344/N rats in NTP studies and the debate about 

whether the NTP will have to go to a more expensive 

and technically difficult dietary restriction regimen for 

all of its studies. Dr. G.N. Rao, NIEHS, said that the 

key to stopping or reversing the upward drift of 

animal body weights is to go back to the production 
D~e~ry Restriction, NT~ T~ 460 

colonies or to establish a colony, to effect controls 

over growth patterns. Dr. A. Turturro, NCTR, 

observed that breeding back will not necessarily yield 

the same animal. He said the large variability 

between individual animals within studies must somehow be controlled or reduced. Dr. W.T. Allaben, 

NCTR, reported that as an outcome of a conference in 

1994, the FDA has put together a draft white paper 

looking at the issue of diet, variability of test outcomes, and the value of caloric restriction in controlling for that variability; this document will soon be 

presented for public comment. Dr. Kari said it is 

important that false negatives and false positives are 

~5 

not masked. Returning to the concept of a workshop, 

Dr. Lucier commented that the impact of dietary 

restriction on additional toxicologic endpoints needs to 

be addressed. Dr. Hart said that the term "dietary 

control" might be preferable to "dietary restriction." 

He said the. FDA would cosponsor a workshop, and 

Dr. Karol indicated that the Society of Toxicology 

would be interested in serving as a cosponsor. 

Dr. Keenan said the Society of Toxicologic Pathologists was planning a symposium in June 1996, and he 

suggested that better integration among sponsoring 

groups was needed. 

I 
16 Dietary Restriction, NTP TR 460 

~ i 
i 

~.7 

! 

INT ©DI.JCT I©N 

It is well documented that dietary restriction 

with concomitant body weight reduction significantly 

increases longevity and decreases the incidence of 

background and chemical-, physical-, and biologicalinduced tumors in rats and mice (Tannenbaum, 1940; 

Ross and Bras, 1973; Gross and Dreyfuss, 1984; 

Pollard et al., 1984; Weindruch and Walford, 1988). 

The interrelationships between body weights, survival 

rates, and neoplasm incidences suggest that practical 

benefits as well as problematic confounding factors 

may be introduced when feed intake, body weights, or 

both are intentionally or unintentionally altered in 

toxicity and carcinogenicity bioassays for chemical 

hazard identification. 

Typically, chronic studies involve feeding rodents 

ad libitum while exposing them to several concentrations of a chemical for up to 2 years. In dietary 

restriction paradigms, the feed presented to control 

and dosed animals is restricted to amounts that result 

in reduced body weights relative to those of animals 

fed ad libitum. Because diet restriction fosters leaner 

animals that live longer than more obese animals 

(Maeda et al., 1985; Yu et al., 1985), experiments 

conducted with dietary restriction may permit higher 

survival rates, thereby allowing more opportunities for 

chemical exposure and more time for treatmentrelated lesionsto develop. It follows that these 

influences might enhance statistical power and 

increase the ability to resolve chemical effects in 

toxicity and carcinogenesis studies. 

These potentially beneficial influences could be 

confounded by the propensity of dietary restriction to 

generally decrease tumor incidence.. A chemical 

observed to be toxic or carcinogenic in animals fed 

ad libitum might not produce the same effects in 

diet-restricted or otherwise leaner animals. This 

discordance could cause difficulties in comparing 

tumor outcomes within and between studies. For 

example, observations collected for over a decade 

from the NTP show that some background tumor rates 

have comigrated with increasing body weights 

(Haseman and Rao, 1992; Haseman, 1993). Theoretically, comparisons between otherwise identical studies 

conducted several years apart could yield disparate 

results influenced primarily by the body weight of the 

animals. 

~TUD~ ]~,~AT~ONALE 

Chemical-associated body weight depression in 

13-week toxicity studies is routinely used in conjunction with other factors to select exposures for 2-year 

toxicity and carcinogenicity studies. By design, the 

highest dose concentrations selected for some toxicity 

studies cause body weight depression. Comparisons 

in an experiment in which dose-related decreases in 

feed intake or body weight occur are potentially confounded by the influence of body weight on survival 

and disease processes. As fewer neoplasms would be 

expect6d in the leaner animals than in similarly dosed 

but heavier animals, carcinogenic activity might be 

underestimated. 

To begin assessing the merits and limitations of 

different dietary regimens in bioassays, studies were 

undertaken to compare the outcomes of four chemicals 

evaluated under typical NTP bioassay conditions as 

well as under protocols including dietary restriction. 

Specifically, these experiments were designed to 

evaluate the effect of dietary restriction (to achieve a 

body weight reduction of approximately 15 % compared to animals fed ad libitum) and to evaluate the 

effect of weight-matched control groups on the 

sensitivity of the bioassays. The results, based on 

these data, show a marked difference in bioassay 

outcome depending upon the protocol used. 

Studies were designed to compare the toxicity and 

carcinogenicity of four chemicals, each evaluated 

under three (t-butylhydroquinone study only) or four 

different protocols. In each protocol, the effects of 

the chemical were assessed by a comparison between 

a group administered a single concentration of the 

chemical and a control group (Figure 1). The 

i 

i 

i 

i 
18 Dietary Restriction, NTP TR 460 

BIOAS SAY PROTOCOL 

AD LIBITUM-FED (104 WEEKS) 

Control 

114 X 

1/2x. 

" lx 

Weight Matched Control 

FEED RESTRICTED (104 WEEKS) 

Control 

lX 

# ANIMALS 

60 

60 

60 

60~ 

60_ 

60-

60--

COMPARISON . 

. 

2 

I " 

FEED RESTRICTED (157 WEEKS or 20% SURVIVAL) 

Control 50 • I lx 50= 

FIGURE 1 

General Design of Feed Restriction Studies 

concentrations were chosen based upon outcomes of 

13-week studies conducted under ad libitum feeding 

protocols. 

For the core bioassay, groups of about 60 animals, 

fed ad libitum, were allotted to control and dosed 

groups (nominally 0, 1/4X, 1/2X, and IX) for 

approximately 104 weeks or up.to 128 weeks (t-butylhydroquinone study). The comparison between the 

control group and the group receiving the highest dose 

(1X) was used to represent the outcome of the bioassay under ad libitum feeding protocols (Figure 1, 

Comparison 1). 

In asecond experiment, the 1X group was instead 

compared with a weight-matched group of 

60 untreated controls (Figure 1, Comparison 2). The 

daily feed allotment for this ContrOl group was 

restricted so that the mean body weight matched the 

mean body weight of the 1X group. 

Two additional groups of about 60 animals (one 

control and one treated group) were offered feed in 

amounts that limited the mean body weight of the 

control group to approximately 85%~that of tlie 

controls fed ad libitum under the first protocol. 

Animals assigned to this third dietary restriction 

paradigm were evaluated after 104 weeks (Figure 1, 

Comparison 3)... 

Because animals fed a restricted diet are expected to 

live longer than animals fed ad libitum, concurrent 

evaluations could result in comparisons at disproportionate times in their respective lifespans, thereby 
Dietary ~estriction, NTP TR ~60 

masking age-dependent effects. Therefore, a fourth 

protocol was employed to evaluate the influence of an 

additional year of chemical exposure on the neoplasm 

profile of animals fed a restricted diet. Two groups of 

50 animals (one control and one dOsed group) 

received restricted diets, as under the third protocol, 

for 3 years or until survival in either group was 

reduced to 20% (Figure 1, Comparison 4). 

The protocols described above and shown in Figure 1 

were used in the evaluation of four chemicals, butyl 

benzyl phthalate, salicylazosulfapyridine, t-butylhydroquinone, and scopolamine hydrobromide trihydrate. Dietary-restriction protocols were not 

imposed across all dose groups of the four 

gender/species combinations typically used by the 

NTP; however, male and female rats and mice are 

represented in these studies, as are the two principal 

modes of oral exposure (dosed feed and gavage). 

Butyl benzyl phthalate was administered to male and 

female rats in feed. t-Butylhydroquinone was administered to F1 male and female rats in feed; in this 

study, rats were exposed in utero and during lactation 

through F0 (parental) exposure as well as during the 

adult stages of life. Salicylazosulfapyridine was 

administered to male rats and male mice by gavage in 

corn oil. Scopolamine hydrobromide trihydrate was 

administered to male and female mice by gavage in 

~9 

water. For the dosed feed studies, animals were 

offered a single concentration of the chemical 

(IX mg/kg body weight) blended in NIH-07 mashtype diet. For the gavage studies, a single 

concentration of the chemical (1X mg/mL vehicle) 

was mixed in corn oil or distilled water and 

administered to the animals at a dose of 5 mL corn oil 

or distilled water per kilogram body weight (rats) or 

10 mL/kg (mice); control animals received the gavage 

vehicle only. Thus, for feed and gavage exposures, 

animals treated under ad libitum feeding or dietary 

restriction protocols received similar quantities .of a 

given chemical on a body weight basis. 

Butyl benzyl phthalate, t-butylhydroquinone, salicylazosulfapyridine, and scopolamine hydrobromide, 

trihydrate (Figure 2) were administered in feed or by 

gavage for 2 years under the standard NTP ad libitum 

feeding protocol; these studies are presented in NTP 

Technical Reports 445, 457, 458, and 459 (NTP, 

1997a,b,c,d). Additional animal groups were 

included to permit comparisons of the sensitivity of 

assays incorporating ad libitum feeding, matching the 

mean body weights of controls to those of dosed 

animals, and restricting the diet to achieve body 

weights that were approximately 85 % those of controis fed ad libitum. The results of these comparisons 

are presented in this Technical Report. 
20 Dietary Restriction, NTP TR 460 

,: 

. . ,, .' ;:,~, 

CH 3 OH O 

II ~ 1

• C--O--C H2CH2C H2CHz ~ 

. .:. C H3 ..[~r,n~J 

C--O--C H2----~ ~ i ~ ~ • \.__i . OH

U 

BUTYL BENZYL PHTHALATE t-BUTYLHYDROQUINONE 

CAS No. 85-68-7 CAS No. 1948-33-0 

Chemical Formula: C~I-i~200'4 Chemical Formula: CtoH1402 

• Molecular Weight: 312.39 Molecular Weight:- 166.22 . 

., ̧ 

.. 

., .. 

COOH 

CH~OH ~ H. 

,

/]~/OH ~--~ , '~ H./--~---i'.. ' 

(' '~'~C--C-O'~-~ N_CH3 I .O • HBr .3H20 N ~ - ~ - ~ ~_~,_.:_~, • I ~ \--/ , ~.~/ .. N N 

o 

. 

SALICYLAZOSULFAPYRIDINE SCOPOLAMINE HYDROBROMIDE 

' CAS No. 599-79-1 TRIHYDRATE 

Chemical Formula: C~sH~N4OsS CAS No. 6533-68-2 -. Molecular Weight: 398.39 Chemical Formula: C17H~iNO4oHBro3H20 

Molecular Weight: 438.31 

FIGURE 2 

Chemical Structures and Information for Compounds Used to Determinethe Effects of Dietary Restriction 

on To:Neology and Carcinogenesis Studies in Rats and Mice .. 

~! . ~ 
21 

M TE I LS 

PROCUREMENT 

AND CHARACTERIZATION 

Butyl benzyl phthalate was obtained from Chem 

Central (Kansas City, MO). t-Butylhydroquinone was 

obtained from U.O.P., Inc. (Des Plaines, IL). 

Salicylazosulfapyridine was obtained from Pharmacia, 

Inc. (Piscataway, NJ). Scopolamine hydrobromide 

trihydrate was obtained from Henley and Company, 

Inc. (New York, NY). Identity, purity, and stability 

analyses were conducted by the analytical chemistry 

laboratory, Midwest Research Institute (Kansas City, 

MO). Reports on analyses performed in support of 

these studies are on file at the National Institute of 

Environmental Health Sciences (NIEHS). Detailed information on the analyses of each chemical is 

also provided in the Technical Reports (NTP, 

1997a,b,c,d). 

. , 

The identity of each chemical was confirmed by 

infrared, ultraviolet, and nuclear magnetic resonance 

spectroscopy. The purity of each chemical was 

determined by elemental analysis, Karl Fischer water 

analysis, functional group titrations, and one or more 

chromatographic methods. Butyl benzyl phthalate and 

t-butylhydroquinone were each determined to have a 

purity of approximately 99 %. Salicylazosulfapyridine 

was determined to have a purity of at' least 98%. 

Scopolamine hydrobromide trihydrate was determined 

to have a purity of approximately 89% (with 11% 

water). The stability of each bulk chemical was 

monitored by the study laboratory; no degradation of 

any of the bulk chemicals was detected. ' 

~REPARA'I'~ON AND ANAL'~'SIS 

OF DOSE ~'ORMULAT~ON$ 

Detailed information on dose preparation and analyses 

for butyl benzyl phthalate, t-butylhydroquinone, 

salicylazosulfapyridine, and scopolamine hydrobromide trihydrate are provided in the Technical Reports 

(NTP, 1997a,b,c,d). For the butyl benzyl phthalate 

and t-butylhydroquinone studies, the bulk chemical 

M ETH© S 

was mixed with feed. For the salicylazosulfapyridine 

studies, the bulk chemical was mixed with corn oil; 

scopolamine hydrobromide trihydrate was mixed with 

distilled Water. , 

Dose formulations for each chemical were periodically analyzed by the study laboratories. Dose 

formulations were generally within 10% of the target 

concentration. Results of periodic referee analyses 

performed by the analytical chemistry laboratory 

agreed with the results obtained by the study laboratories. Periodic analyses by the study laboratory of 

the corn oil vehicle used in the salicylazosulfapyridine 

studies demonstrated that peroxide levels were generally within the acceptable limit of 3 mEq/kg; two 

lots that were slightly outside the acceptable limit 

were replaced as quickly as possible. 

~TUDY DESIGNS 

Diets and ~'eeding 

NIH-07 open formula diet (4.0126 cal/g), obtained 

from Zeigler Brothers, Inc. (Gardners, PA) was 

offered either ad libitum or in restricted quantities. 

For the butyl benzyl phthalate and t-butylhydroquinone feed studies, a mash formulation was used; 

for the salicylazosulfapyridine and scopolamine 

hydrobromide trihydrate gavage studies, feed was 

processed into precise 1-gram pellets, thereby facilitating the presentation of restricted amounts Of feed by 

eliminating the requirement for weighing. 

For the core ad libitum feeding protocol (Figure 1, 

Comparison 1), the control and dosed groups were 

allowed unlimited access to feed. Apparent feed 

intake, as measured by disappearance of feed from 

feed hoppers (cage averages), was generally measured 

; weekly for the first 26 weeks and monthly thereafter. 

In all studies, individual animal body. weights were 

determined weekly for the first 13 weeks and monthly 

thereafter. Feed spillage was not determined; therefore, values reported for feed intake represent uncorrected indexes of actual consumption. 
22 

The daily feed allotment for the weight-matched 

control groups was restricted so that the mean body 

weights of these groups would match those of the 

corresponding 1X groups (Figure 1, Comparison 2). 

The first 2 weeks of each study were used to establish 

baseline averages for body weights and feed intake; 

consequently, dietary restriction was not imposed 

. during this p6riod. Thereafter, daily feed offerings to 

the weight-matched control groups were determined 

f~om the ratio of the mean body weight of dosed 

iinimals ~o that of the ad libitum controls multiplied by 

'the feed intake of the ad libitum controls. For 

example, if the mean body weight of the. 1x group 

was 90% that of the ad libitum controls at a given 

weighing period, the amount of feed presented to the 

weight-matc.hed controls the.following week was 90 % 

that of the average feed intake of the ad libitum 

controls during the week of the given weighing. The 

new weekly target value for feed intake was then 

divi_ded by seven to obtain a daily value, then multiplied by the number of animals per cage and rounded 

to the nearest gram. The resultant feed mass was put 

" into the cage feed hoppers daily. Corrections for 

body weight changes, if necessary, were made weekly 

for the first 13 weeks and monthly thereafter. Corrections for changes in the number of rats per cage due 

to mortality were made as these changes occurred. 

In an analogous manner~ feed-restricted control 

groups (Figure 1, Comparisons 3 and 4) Were offered 

a quantity of feed sufficient to produce a 15 % reductiOfi in body weight compared to the ~ad libitum 

controls. Feed-restricted, dosed .groups were offered 

an amount of feed, identical to that given the feed- 

. restricted controls. This portion of each study was 

begun 2 weeks after the ad libitum feeding portion so 

that baseline values for body weights and feed intake 

could be established. 

k . 

Mean body weight at 1 year was used as an indicator 

of the growth of the animals. By this time, the 

animals generally had had sufficient time to be influenced by adverse effects of the chemical but not to be 

greatly influenced by the variety of changes associated 

with morbidity and mortality in older animals 

(Turturro et al., 1993). 

Dietary Restriction, NTP TR 460 

Butyl Benzyl Phthalate Study 

Groups of 60 male rats were fed diets containing 0 or 

12,000 .ppm butyl benzyl phthalate; groups, of 

60 female rats were fed diets containing 0 or 

24,000 ppm butyl benzyl phthalate. Ten male and 

ten female rats from each group were evaluated at 

15 months for histopathology and organ weights. 

Male and female F344/N rats were obtained from 

Simonsen Laboratories (Gilroy, CA) for use in the 

dietary restriction study. Rats were quarantined for 

10 or 11 days before the beginning of the study. 

Prior to the beginning of treatment,, five male and five 

female rats were selected for parasite evaluation and 

gross observation of disease. Serology samples were 

collected for viral screening. Rats were approximately 6 weeks old at the beginning of the studies. 

The health of the animals was monitored during the 

studies according to the protocols of the NTP Sentinel 

Animal Program. 

Rats were housed five per cage. Water was available 

ad libitum. Cages and racks were rotated, every 

2 weeks. Further details of animal maintenance are . . . . 

given in Table 1. 

t-Butylhydroquinone Study 

The design of the t-buty!hydroquinone study is similar 

to the ~design ..of the butyl benzyl phthalate study, 

except that 70 male and 70 female rats were used and 

the exposure began in utero. Five-week-old female 

F344/N rats were. maintained for 5 to 7 weeks and 

were then housed two per cage. At this time, females 

began receiving 0 or 5,000 ppm t-butylhydroquinone 

in feed; after 2 weeks, one male rat was cohoused 

with each pai r of females. When pregnancy was 

ascertained .by vaginal Smear, the females were 

housed individually; females continued to receive the 

same diet during pregnancy and through weaning of 

the pups..The litters were culled to four males and 

four females on day 4 postpartum. During the fourth 

week postpartum (from day 28 to day 35), two males 

and two females were selected aLrandom from each 

litter until 70 males and 70 females per exposure 

concentration were ~selected. These rats received the 
~

[ 

Dietary Restriction, NT~ T~ 460 

same concentration of t-butylhydroquinone as their 

dams had received for up to 130 weeks. At 3 months, 

10 males and 10 females per group were evaluated for 

histopathology and organ weights. 

Male and female F344/N rats used for breeding were 

obtained from Taconic Laboratory (Germantown, NY) 

for use in the dietary restriction study. Rats were 

quarantined for 18 days before the beginning of the 

study. Prior to the beginning of treatment, 10 male 

and 10 female rats were selected for parasite evaluation and gross observation of disease. Serology 

samples were collected for viral screening. The 

health of the animals was monitored during the studies 

according to the protocols of the NTP Sentinel Animal 

Program. 

F~ rats were housed five per cage. Water was avaiiable ad libitum. Cages and racks were. rotated every 

2 weeks. Further details of animal maintenance are 

given in Table 1. 

Sa~cy~azosu~fapyr~d~ne Studies 

Groups of 60 male rats received 0 or 337.5 mg 

salicylazosulfapyridine per kilogram body weight in 

corn oil by gavage. Groups of 60 male mice received 

0 or 2,700 mg/kg salicylazosulfapyridine in corn oil 

by gavage. Ten rats and ten mice from each group 

were evaluated at 15 months for histopathology and 

organ weights. 

Male F344/N rats and B6C3F1 mice were obtained 

from Simonsen Laboratories (Gilroy, CA) for use in 

the dietary restriction studies. Rats and mice were 

quarantined for 11 to 13 days before the beginning of 

the studies. Prior to the beginning of treatment, five 

rats and five mice were selected for parasite evaluation and gross observation of disease. Serology 

samples were collected for viral screening. Rats and 

mice were approximately 6 to 7 weeks old at the 

beginning of the studies. The health of the animals 

was monitored during the studies according to the 

protocols of the NTP Sentinel Animal Program. 

Rats were housed five per cage and mice were housed 

individually. Water was available ad libitum. Cages 

and racks were rotated every 2 weeks. Further details 

of animal maintenance are given in Table 1. 

Scopolamine Hydlrobrom~e Tr~lhydrate 

Study 

Groups of up to 70 male and 70 female mice received 

0 or 25 mg scopolamine hydrobromide trihydrate per 

kilogram body weight in deionized water by gavage. 

(Ten males and nine females in each group fed 

ad libitum were evaluated for scopolamine hydrobromide trihydrate levels in plasma and were discarded without further evaluation; results of these 

analyses are included in NTP, 1997a). Ten male and 

ten female mice from each group were evaluated at 

15 months for histopathology and organ weights. 

Male and female B6C3F1 mice were obtained from 

Simonsen Laboratories (Gilroy, CA) for use in the 

2-year studies. Mice were quarantined, for 14 days 

before the beginning of the studies. Prior to the 

beginning of treatment, five male and five female 

mice were selected for parasite evaluation and gross 

observation of disease. Serology samples were 

collected for viral screening. Mice were approximately 6 weeks old at the beginning of the studies. 

The health of the animals was monitored during the 

studies according to the protocols of the NTP Sentinel 

Animal Program. 

Mice were housed individually. Water was available 

ad libitum. Cages and racks were rotated every 

2 weeks. Further details of animal maintenance are 

given in Table 1. • 

C~n~cz]l E~n~nzfions ~n~l ~atho~o~y 

All animals were observed twice daily. Clinical" 

findings and body weights were recorded as described 

in Table 1 . 

A complete necropsy and microscopic examination 

were performed on rats and mice. At each interim 

evaluation necropsy, selected organs of rats and mice 

were weighed (see Table 1). At necropsy, all organs 

and tissues were examined for grossly visible lesions, 

and all major tissues were fixed and preserved in 10% 

neutral buffered formalin, processed and trimmed, 

embedded in paraffin, sectioned to a thickness of 5 to 

6 /~m, and stained with hematoxylin and eosin for 

microscopic examination. For all paired organs (i.e., 

.adrenal gland, kidney, ovary), samples from each 

organ were examined. Tissues examined microscopically are listed in Table 1. 
24 

. Microscopic evaluations:were completed by the study 

laboratory pathologist, and the pathology data were 

entered into the Toxicology Data Management 

System. The slides, paraffin blocks, and residual wet ~f . 

t~ssues were sent to the NTP Archives for inventory, 

slide/block match, and wet tissue audit. Theslides, 

individual animal data records, and pathology tables 

were evaluated by an independent quality assessment 

laboratory. The individual animal records and tables 

were compared for accuracy, the .slide and tissue 

"counts were verified, and the histotechnique was 

evaluated. A quality assessment pathologist reviewed 

selected organs in each study. In the butyl benzyl • 

phthalate study, the organs examined included the 

adrenal gland, .kidney, liver, mammary gland (fe- ! ma)es), pancreas, pituitary gland (pars distalis), 

.~spleen, thyroid gland (males), urinary bladder, and 

~ uterus. In the. t-butylhydroquinone study, the organs 

examined included the bone marrow (females), 

clitoral gland, forestomach (males), kidney (males), 

liver; mammary gland, nose, pituitary gland, preputial 

gland, spleen~ and thyroid gland (males). In. the 

salicylazosulfapyridine studies, selected neoplasms 

and nonneoplastic lesions were examined. In the 

scopolamine hydrobromide trihydrate study, the 

organs and tissu_es examined included the bone 

marrow (females), forestomach, liver, lung, kidney, 

pancreatic islets (males), pituitary gland (females), 

preputial gland, teeth (males), thyroid gland, and 

uterus. • 

• 

The quality assessment report and the reviewed slides 

: 

were submitted to the NTP Pathology. Working Group 

(PWG) chairperson, who reviewed the selected tissues 

and addressed any inconsistencies in the diagnoses 

made by the laboratory and qual!ty assessment patho!- 

ogists. Representative .histopathology slides containing examples Of lesions related to chemical 

:administration, examples of disagreements in diagnoses between the laboratory and quality assessment 

pathologists, or lesions of general interest were 

presented by the chairperson to the-PWG for review. 

The PWG consisted of the quality assessment pathologist and other pathologists experienced in rodent 

toxicologic pathology. This groupexamined the 

tissues without any knowledge of dose. groups or 

previously rendered diagnoses. When the PWG 

consensus differed from the opinion of the laboratory 

pathologist, the diagnosis was changed. Thus, the 

final diagnoses represent a consensus of quality 

Dietary Restriction, NTP TR 460 

assessment pathologists, .the PWG chairperson; and 

the PWG. Details of~these review procedures have 

been described, in part, by Maronpot and Boormhn 

(1982) and Boorman et al. (1985). For subsequent 

analyses Of the pathology data, the diagnosed lesions 

for each tissue type were evaluated separately or 

combinedaccording, to the guidelines of McConnell 

e~ al. (1986). • " • 

STATISTICAL METHODS • 

Survival Analyses : 

The probability of survival'was estimated by. the 

product-limit procedure of Kaplan and Meier (1958) 

and is presented in the form of graphs. Animals 

found dead of other than natural causes• or missing 

were censored from the survival analyses; animals 

dying from natural causes were not censored. Statistical analyses for POssible chemical-related effects .on 

survival used Cox's (!972) method for testing two 

groups for equality and Tarone' s (1975) life table test. 

All reported P values for the survival analy~es are two 

sided. . .. 

Calculation of Incidence 

The incidences of neoplasms or nonneoplastic lesions 

as presented in Tables A1, A3, B1, B3, C!, C3, D1, 

D3, El, E3, Fi,'F3, G1, G3, H1, and H3 are.given 

as the number of animals bearing such lesions at a 

specific anatomic site and the number of animals with 

that site exarn~ned microscopically. For calculation.of 

statistical significance, the incidences of most ne0- 

p.lasms (Tables A2, B2, C2, D2, E2, F2, G2, and H2) 

and all nonneoplastic lesions are given as the numbers 

of animals affected at each site examined mici~oscopically. However, when macroscopic examination was 

required to detect neoplasms in certain tissues (e.g., 

skin; intestine, harderian gland, and mammary gland) 

before microscoPic evaluation, or when neoplasms 

had multiple potential sites of occurrence (e.g., 

leukemia or iymphoma), the denominators consist of 

the number of animals On which, a necropsy was 

performed, Tables A2, B2, C2, D2,' E2, F2, G2, and 

H2 also give the survival-adjusted tumor rate. for each 

group and each site-specific tumor, i.e., the KaplanMeier estimate of the tumor incidence that would have 

been observed at the end of the study in the absence 

of mortality from all other competing risks (Kaplan 

and Meier, 1958). . 
Dietary Restriction, NTP TR 460 25 

Analysis of Neoplasm Incidences 

The majority of neoplasms in these studies were 

considered to be incidental to the cause of death or not 

rapidly lethal. Thus, the primary statistical method 

used was logistic regression analysis, which assumed 

that the diagnosed neoplasms were discovered as the 

result of death from an unrelated cause and thus did 

not affect the risk of death. In this approach, neoplasm prevalence was modeled as a logistic function 

of chemical exposure and time. Both linear and 

quadratic terms in time were incorporated initially, 

and the quadratic term was eliminated if the fit of the 

model was not significantly enhanced. The neoplasm 

incidences of exposed and control groups were 

compared on the basis of the likelihood score test for 

the regression coefficient of dose. This method of 

adjusting for intercurrent mortality is the prevalence 

analysis of Dinse and Lagakos (1983), further 

described and illustrated by Dinse and Haseman 

(1986). When neoplasms are incidental, this comparison of the time-specific neoplasm prevalences also 

provides a comparison of the time-specific neoplasm 

incidences (McKnight and Crowley, 1984). 

In addition to logistic regression, other methods of 

statistical analysis were used, and the results of these 

tests are summarized in the appendixes. These 

methods include the life table test (Cox, 1972; 

Tarone, 1975), appropriate for rapidly lethal neoplasm s, and the Fisher exact test (Gart et al., 1979), 

a procedure based on the overall proportion of 

neoplasm-bearing animals. . ,. 

Tests of significance included pairwise comparisons of 

each exposed group with controls. • Continuity=corrected tests were used in the analysis of neoplasm 

incidence, and reported P values are one sided. The 

procedures described in the preceding paragraphs 

were also used to evaluate selected nonneoplastic 

lesions. For further discussion of these statistical 

methods, refer to Haseman (1984). 

An~ysis of Nonneop~astic Lesion l~ncidences 

Because all nonneoplastic lesions in this study were 

considered to be incidental to the cause of death or not 

rapidly lethal, the primary statistical analysis used was 

a logistic regression analysis in which nonneoplastic 

lesion prevalence was modeled as a logistic function 

of chemical exposure and time. For lesions detected 

at the interim evaluation of all studies and for lesions 

detected in animals treated under the restricted-feed 

protocols, the Fisher exact test, a procedure based on 

the overall proportion of affected animals, was used. 

.. 

Araa~ysis of Continuous Variables 

Organ and body weight data, which have approximately normal distributions, were analyzed with 

Student's t-test. 

Historical Control Data " 

Although the concurrent control group is always the 

first and most appropriate control group used for 

evaluation, historical control data can be helpful in the 

overall assessment of neoplasm incidence in certain 

instances. Consequently, neoplasm incidences from 

the NTP historical control database, which is updated 

yearly, are included in the NTP reports for neoplasms 

appearing to show compound-related effects. 

QUALI~TY ASSURANCE i~E'~HODB 

The studies were conducted in compliance with Food 

and Drug Administration Good Laboratory Practice 

Regulations (21 CFR, Part 58). In addition, as 

records from the studies were submitted to the NTP 

Archives, these studies were audited retrospectively 

by an independent quality assurance contractor. 

Separate audits covering completeness and accuracy 

of the pathology data, pathology specimens, final 

pathology tables, and a draft of this NTP Technical 

Report were conducted. Audit procedures and findings are presented in the reports and are on file at 

NIEHS. The audit findings were reviewed and 

assessed by NTP staff, so all comments had been 

resolved or were otherwise addressed during the 

preparation of this Technical Report. 
26 Dietary Restriction, NTP TR 460 ., 

TABLE 1 

Experimental Design and Materials and Methods in the Study of the Effect of Dietary Restriction 

on Toxicology and Carcinogenesis Studies in F344/N Rats and B6C3FI Mice 

Butyl Benzyl t-Butylhydroquinone Salicylazo- scopolamine 

Phthalate sulfapyridine Hydrobromide Trihydrate 

Study Laboratory 

Southern Research Institute 

(Birmingham, AL) 

~.~ ~ Strain and ~pecms ,, 

Rats: F344/N 

Animal Source 

Simonsen Laboratories, ~ 

Inc. (Gilroy, CA) 

Time Held Before Studies 

10-11 days :. ~ 

• 

SouthernResearch Institute 

(Birmingham, AL) 

Rats: F344/N' 

Fo: Taconic Laboratory 

(Germantown, NY) 

Fj: Bred at study 

laboratory 

Fo: 18 days 

Average Age When Studies Began 

6 weeks F~ ad libitum-fed and 

weight-matched: 

5 weeks 

Fj feed-restricted: 

5 weeks .. 

Date of First Dose 

Ad libiturn-fed and F. ad libitum-fed and 

weight-matched: weight-matcbed: 

26 June 1989 29 January 1990 

Feed-restricted: F~ feed-restricted: 

17 July 1989 12 February 1990 

Duration of Dosing 

Ad libitum-fed and Fi ad libitum-fed and 

weight-matched: weight-matched: 

105 (male) or 106 weeks 122 (male) or 128 weeks 

(female) (7 days/week) (female) (7 days/week) 

Feed-restricted: • Fi feed-restricted: 

105 weeks (male and 130 weeks (7 days/week) 

female), 128 weeks 

(male), or 140 weeks 

(femalej (7 days/week) 

Southern Research Institute 

(Birmingham, AL) 

Rats: F344/N males 

Mice: B6C3Fi males 

Simonsen Laboratories, 

Inc. (Gilroy, CA) 

Rats: 11-12days 

Mice: 11-13 days 

6 to 7 weeks 

Ad libitum-fed and 

weight-matched: 

12 December 1988 (rats) 

3 January 1989 (mice) 

Feed-restricted: 

27 December 1988 (rats) 

16 January 1989 (mice) 

Ad libitum-fed and 

weight-matched: 

103 weeks (5 days/week) 

Feed-restricted: 

104, 130 (rats.), or 

156 weeks (mice) 

(5 days/week) 

Battelle Columbus 

(Columbus, OH) 

Mice: B6C3Fl 

Simonsen Laboratories, 

Inc. (Gilroy, CA) 

14 days 

6 weeks 

. 

Ad libitum-fed and 

weight-matched: 

22 September 1988 

Feed-restricted: 

6 October 1988 

Ad libitum-fed and 

weight-matched: 

104 weeks (5 days/week) 

Feed-restricted: 

104 or 157 ~eeks 

(5 days/week) 

.. 
Dietary Restriction, NTP TR 4~0 27 

TAnLE 1 

Exl~er~ment~fl Design and 1V~aterials and Methods ~n the Study of the Effect of Dietary Restriction 

on To~fico~ogy znd Carcinogenesis Studies ~n 1~'344fN Rats and ~6C31~' t Mice (continued) 

Butyl Benzyl 

Phthalate 

Necropsy Dates 

Ad libitum-fed and 

weight-matched: 

15-Month interim 

evaluation - 

26-27 September 1990 

Termination - 

24-26 June (male) and 

1-3 July (female) 1991 

Feed-restricted: 

15-Month interim 

evaluation - 

16 October 1990 

2-Year termination - 

15-17 July 1991 

30/32-Month 

termination - 

30 December 1991 

(male) and 

19 March 1992 (female) 

Average Age at Necropsy 

Ad libitum-fed and 

weight-matched: 

110 weeks (male); 

111 weeks (female) 

Feed-restricted: 

110 or 134 weeks 

(male); 

110 or 145 weeks 

(female) 

Size of Study Groups 

15-Monlh interim 

evaluation - 

10 males and 10 females 

Termination -

50 males and 50 females 

t-Butylhydroquinone 

Ft ad libitum-fed and 

weight-matched: 

3-Month interim 

evaluation - 

30 April (male) and 

1 May (female) 1990 

Termination3 June (male) and 

14-15 July (female) 1992 

F~ feed-restricted: 

3-Momh interim 

evaluation - 

14 May 1990 

Termination - 

10-12 August 1992 

F~ ad libitum-fed and 

weight-matched: 

127 weeks (male); 

133 weeks (female) 

Ft feed-restricted: 

135 weeks 

F~: 3-Month interim 

evaluation - 

10 males and 10 females 

F~: Termination60 males and 60 females 

Salicylazosulfapyridine 

Ad libitum-fed and 

weight-matched: 

15-Month interim 

evaluation - 

12-13 March 1990 (rats) 

3 April 1990 (mice) 

Termination - 

10-14 December 1990 

(rats) 

31 December 1990- 

4 January 1991 (mice) 

Feed-restricted: 

15-Month interim 

evaluation - 

27 March 1990 (rats) 

17 April 1990 (mice) 

2-Year termination - 

26-27 December 1990 

(rats) 

14-15 January 1991 

(mice) 

30-Month/3-year 

termination - 

24 June 1991 (rats) 

13 January 1992 (mice) 

Ad libitum-fed and 

weight-matched: 

110-112 weeks 

Feed-restricted: 

110 or 136 weeks (rats) 

110 or 162 weeks (mice) 

15-Month interim 

evaluation - 

10 males 

Termination - 

50 males 

Scopolamine 

Hydrobromide Trihydrate 

Ad libitum-fed and 

weight-matched: 

15-Month interim 

evaluation - 

21-22 December 1989 

Termination - 

17-20 September (male) 

and 17-21 September 

(female) 1990 

Feed-restricted: 

15-Month interim 

evaluation - 

4 January 1990 

2-Year termination - 

27-28 September 1990 

3-Year termination - 

26-27 September 1991 

Ad libitum-fed and 

weight-matched: 

110 weeks 

Feed-restricted: 

110 or 162 weeks 

15-Month interim 

evaluation - 

10 males and 10 females 

Termination - 

50 males and 50 females 
28 Dietary Restriction, NTP TR 4(~0 

TABLE 1 

Experimental Design' and Materials and Methods in .the Study of the Effect. of Dietary Restriction 

on Toxicology and Carcinogenesis Studies in F344/N Rats and B6C3Fi Mice (continued) • 

Butyl Benzyl t-Butylhydroquinone 

Phthalate 

Method of Distribution 

Animals Were distributed Same as butyl benzyl 

randomly into groups of phthalate study ~ • . 1 . . . approx~mately~equal ~mt~al 

mean body. weight. 

Animals per Cage 

5 F~: 5 : 

. 

Method of Animal Identification 

Tail tattoo - Tail tattoo 

Diet 

NIH-07 open formula meal Same as butyl benzyl 

diet (Zeigler Brothers, phthalate study 

Inc., Gardners, PA), 

available ad libitum, 

changed daffy 

Water Distribution 

Tap water (Birmingham Same as butyl benzyl 

municipal supply) via phthalate study. 

automatic Watering system. 

(Edstrom Industries, 

Waterford, WI), available 

ad libitum 

~ , , 

Cages 

Polycarbonate (Lab Same as buty! benzyl 

Products, Inc., Maywood, phthalate study 

NJ), changed twice weekly 

Bedding 

Sani-Chips ® hardwood Same as butyl bgnzyl 

chips (P.J. Murphy Forest phthalate study 

Product~, Montville, NJ), 

changed twice weekly 

Salicylazo~ 

sulfapyridine 

Same as butyl benzyl " 

phthalate study 

Rats: 5 

Mice: 1 

Rats: Tail tattoo 

Mice: Toe clip 

Same as butyl benzyl 

phthalate study, changed 

twice weekly 

Same as bmyl benzy! 

phthalate study 

Same as butyl benzyl 

phthalate study, except 

changed weekly for mice 

Same as butyl benzyl 

phthalate study, except 

changed weekly for mice 

Scopolamine 

Hydrobromide Trihydrate 

Same as butyl benzyl 

phthalate study 

1 

Toe clip 

NIH-07 open formula 

pellets (Zeigler Brothers, 

Inc., Gardners, PA), 

available ad libitum and 

changed weekly,-or 1-gram 

pellets, available in fixed 

amounts and changed daily 

Tap water (Columbus 

municipal supply) via 

automatic watering system 

(Edstrom Industries, 

Waterford, Wi), gvailable 

ad libitum 

Polycarbonate (Lab 

Products, Inc., Maywood, 

NJ), rotated.every 2 weeks 

Same as butyl benzyl 

phthalate study, except. 

changed weekly 
.... =~ ~ = ~ 

Dietary Restriction, NTP TR 460 29 

TABLE ~ ~[ • 

Exper~nental Design ~ Materials ~nd Methods ~n the Study of the Effect of D~et~ry ~estr~c~on 

on To~co~o~ ~d Carc~ogenes~s Stu~es ~ ~ 

Butyl Benzyl t-Butylhydroquinone 

Phthalate 

Cage FFilters 

Remay ® spun-bonded Same as butyl benzyi 

polyester (Andico, phthalate study 

Birmingham, AL), 

changed every 2 weeks 

Racks 

Stainless steel (Lab Same as butyl benzyl 

Products, Inc., Maywood, phthalate study 

NJ), changed every 

2 weeks 

Animal Room Environment 

Ad libitum-fed and Ft ad libitum-fed and 

weight-matched: weight-matched: 

Temperature - Temperature -

13.1" to 29.2 ° C 17.3" ,to 28.8* C 

Relative humidity - Relative humidity - 

22% to 89% 17% to 85% 

Feed-restricted: F~ feed-restricted: 

Temperature - Temperature -

13.3" to 28.6 ° C 14.2" to 26.7* C 

Relative humidity - Relative humidity - 

18% to 91% 19% to 90% 

All groups: All Ft groups: 

Fluorescent light - Fluorescent lighi - 

12 hours/day 12 hours/day 

Room air - Room air -

minimum of minimum of 

10 changes/hour 10 changes/hour 

Doses 

0 or 12,000 ppm (males) 0 or 5,000 ppm in feed, : 

and 0 or 24,000 ppm available ad libitum or in 

(females) in feed that was restricted amounts 

available ad libitum or in 

restricted amounts 

~z~ .~d ~6C3~ ~ce 

Sa!icylazosulfapyridine 

Same as butyl benzyl 

phthalate study 

Same as butyl benzyl 

phthalate study 

Ad libitum-fed and 

weight-matched: 

Temperature -

13.5" to 25.9* C (rats), 

13.4 ° to 26.3* C (mice) 

Relative humidity - 

20% to 90% (rats), 

15% to 87% (mice) 

Feed-restricted: 

Temperature -

16;2" to 29.4* C (rats), 

17.3 ° to 29.7 ° C (mice) 

Relative humidity - 

21% to 90% (rats), 

22% to 85% (mice) 

All groups: 

Fluorescent light - 

12 hours/day 

Room air -

minimum of 

10 changes/hour 

Rats: 0 or 337.5 mg/kg 

body weight in corn oil 

by gavage at a volume of 

5 mL/kg body weight 

Mice: 0 or 2,700 mg/kg 

body weight in corn oil 

by gavage at a volume of 

I0 mL/kg body weight 

(eon~u~) .~ 

Scopolamine. , 

Bydrobromide Trihydrate 

DuPont 2024 spun-bonded 

polyester (Snow Filtration, 

Cincinnati, OH), . changed 

every 2 weeks 

Same as butyl benzyl 

phthalate study 

Ad h'bitum-fed and 

weight-matched: 

Temperature -

20.0* to 24.4* C 

Relative humidity - 

30% to 73% 

Feed-restricted: 

Temperature -

18.3" to 26.7* C 

Relative humidity - 

15% to 79% 

All groups: 

Fluorescent light - 

12 hours/day 

Room air -

minimum of 

10 changes/hour 

0 or 25 mg/kg body weight 

in deionized water by 

garage at a volume of 

10 mL/kg body weight 

" i 

.~ : 

_ 

-~ 
30 Dietary Restriction, NTP TR 460 

TABLE 1 

Experimental Design and Materials and Methods in the Study of the Effect of Dietary Restriction 

on Toxicology and Carcinogenesis Studies in F344/NRats and B6C3F1 Mice (continued) 

. 

Butyl Benzyl t-Butylhydroquinone Salicylaz0- Scopolamine 

Phthalate sulfapyridine Hydrobromide Trihydrate 

Type and Frequency of Observation 

Observed twice daily. ' 

Animals were weighed and 

clinical findings were 

recorded initially, weekly 

for 13 weeks, monthly 

thereafter, and at the end 

of the studies. Feed 

consumption by rats fed 

ad libitum was recorded 

weekly by cage for 

26 weeks and monthly 

thereafter; feed 

consumption for other 

groups was recorded daily 

for 15 months and monthly 

thereafter. 

Method of Sacrifice 

C02 asphyxiation 

F~: Observed twice daily. 

Animals were weighed and 

clinical findings were 

recorded initially; on 

lactation days 4, I1, 18, 

and 28; at the beginning of 

adult exposures; weekly 

for 13 weeks; monthly, 

thereafter; and at the end 

of the studies. Feed 

consumption by control 

rats fed ad libitum and 

weight-matched controls 

was recorded weekly by 

cage for 26 weeks and 

monthly thereafter. Feed 

consumption for restrictedfeed groups was recorded 

weekly for 36 weeks and 

monthly thereafter. Feed 

consumption for other 

groups was recorded 

monthly. 

CO2 asphyxiation 

Observed twice daily. 

Animals were weighed and 

clinical findings were 

recorded initially, weekly 

for 13 weeks, monthly 

thereafter, and at the end 

of the studies. Feed 

consumption by control 

rats (by cage) and mice (by 

animal) fed ad libitum was 

recorded weekly for 

26 weeks and monthly 

thereafter. Feed 

consumption by other rat 

groups was recorded 

monthly. Feed 

consumption-by weightmatched and feed restricted 

mouse groups was 

recorded daily for 

21 months, then 5 days a 

week for 1 week each 

month. For dosed mice 

fed ad libitum; feed 

consumption was recorded 

5 days a week for 1 week 

each month. 

COs asphyxiation ~ . • .. 

Observed twice daily. 

Clinical findings were 

recorded initially, monthly 

thereafter, and at the end 

of the studies.- Animals 

were weighed initially, 

weekly for 13 weeks, 

monthly thereafter, and at 

the end of the studies. 

Feed consumption was not 

recorded. 

Anesthetization with 

C02:02 followed by 

exsanguination by cardiac 

puncture (for groups with 

blood analyses); CO2:Oz 

asphyxiation (all other " 

groups) 
Dietary ]Restriction, NT~ TR 460 31 

'FABLE ~. 

Ex~er~menta~ Design ~nd Materials and Methods [n the Study of the Effect of Dietary ~est~ction 

on Tordco~ogy and Carc~ogenesis Studies ~n ~¢3~/N 

Butyl Benzyl t-Butylhydroquinone 

Phthalate 

Necropsy 

Necropsy was performed Necropsy was performed 

on all rats. Organs on all Ft rats. Organs 

weighed at the 15-month weighed at the 3-month 

interim evaluation were interim evaluation were 

right epididymis, right right epididymis, right 

kidney, liver, and right kidney, liver, and right 

testis. testis. 

Histopathology 

Complete histupathology Complete .histopathology 

was performed on all rats. was performed on all rats. 

In addition to gross lesions In addition to gross lesions 

and tissue masses, the and tissue masses, the 

tissues examined included: tissues examined included: 

adrenal gland, bone and adrenal gland, bone and 

marrow, brain, clitoral marrow, brain, clitoral 

gland, epididymis, gland, epididymis, 

esophagus, eyes (if grossly esophagus, eyes (if grossly 

abnormal), heart, kidney, abnormal), heart, kidney, 

large intestine (cecum, large intestine (cecum, 

colon, and rectum), liver, colon, and rectum), liver, 

lung, lymph nodes lung, lymph nodes 

(mandibular and (mandibular and . 

mesenteric), mammary mesenteric), mammary 

gland, nose, ovary, gland, nose, ovary, 

pancreas, parathyroid pancreas, parathyroid 

gland, pharynx (if grossly gland, pharynx (if grossly 

abnormal), pituitary gland, abnormal), pituitary gland, 

preputial gland, prostate preputial gland, prostate 

gland, salivary gland, gland, salivary gland, 

seminal vesicle, spleen, seminal vesicle, spleen, 

small intestine (duodenum, small intestine (duodenum, 

jejunum, and ileum), spinal jejunum, and ileum), spinal 

cord and sciatic nerve (if cord and sciatic nerve (if 

neurologie signs were neurologic signs were 

present), stomach present), stomach 

(forestomaeh and glandular (forestumach and glandular 

stomach), testis, thymus, stomach), testis, thymus, 

thyroid gland, trachea, thyroid gland, trachea, 

Urinary bladder, and urinary bladder, and 

UteruS. UteruS. 

~ats and ~6C3~'~ l~,~ce (continued) 

Salicylazosulfapyridine 

Necropsy was performed 

on all animals. Organs 

weighed at the 15-month 

interim evaluations were 

right kidney, liver, spleen, 

right testis, and right 

thyroid gland. 

Complete histopathology 

was performed on all rats 

and mice. In addition to 

gross lesions and tissue 

masses, the tissues 

examined included: 

adrenal gland, bone and 

marrow, brain, 

epididymis, esophagus, 

eyes (if grossly abnormal), 

gallbladder (mice), heart, 

kidney, large intestine 

(cecum, colon, and 

rectum), liver, lung,, lymph 

nodes (mandibular and ... 

mesenteric), mammary 

gland, nose, pancreas, 

parathyroid gland, pharynx 

(if grossly abnormal), 

pituitary gland, preputial 

gland, prostate gland, 

salivary gland, seminal 

vesicle, spleen, small 

intestine (duodenum, 

jejunum, and ileum), spinal 

cord and sciatic nerve (if 

neurologic signs were 

present), stomach 

(forestomach and glandular 

stomach), testis, thymus, 

thyroid gland, trachea, and 

urinary bladder. 

Scopolamine 

Hydrobromide Trihydrate 

Necropsy was performed 

on all mice. Organs 

weighed at the 15-month 

interim evaluation were 

right epididymis, right 

kidney, liver, and right 

testis. 

Complete histopathology 

was performed on all 

mice. In addition to gross 

lesions and tissue masses, 

the tissues examined 

included: adrenal gland, 

bone and marrow, brain, 

clitoral gland, epididymis, 

esophagus, eyes (if grossly 

abnormal), gallbladder, ~ 

heart, kidney, large 

intestine (cecum, colon, 

and rectum), liver, lung, 

lymph nodes (mandibular 

and mesenteric), mammary 

gland, nose, ovary, 

pancreas; parathyroid 

gland, pharynx (if grossly 

abnormal), pituitary gland, 

preputial gland, prostate 

'gland, salivary gland, 

seminal vesicle, spleen, 

small intestine (duodenum, 

jejunum, and ileum), spinal 

cord and sciatic nerve (if 

neurologic signs were 

present), stomach 

(forestomach and glandular 

stomach), testis, thymus, 

thyroid gland, trachea, 

urinary bladder, and 

uterus. 
32 Dietary Restriction, NTP TR 460 

L . 
33 

 UT fL IFNT ALAT 

AD LIBITUM FEEDING AND W~EGHT-MATCHED CONTROLS ]P~;~OTOCOL~ 

Survival 

Estimates of 2-year survival probabilities for male and 

female rats fed ad libitum and the weight-matched 

control rats are shown in Table 2 and in the KaplanMeier survival curves in Figure 3. The survival rates 

of exposed male and female rats were similar to those 

of the controls fed ad libitum but less than those of the 

weight-matched controls. 

~ody Weights 

Mean body weights are given in Figure 4 and in 

Appendix. J, Table J1. At 1 year, the mean body 

weight of exposed males was approximately 92 % that 

of the controls fed ad libitum and 104% that of the 

weight-matched controls~ The mean body weight of 

females in the 24,000 ppm group was 80% that of the 

controls fed ad libitum and 108% that of the weightmatched controls at 1 year. From 1 year through the 

end of the study, the mean body weight of exposed 

males remained within 8 % that of the controls fed 

ad libitum and within 7 % that of the weight-matched 

controls; the mean body weight of exposed females 

fell to 73 % that of the controls fed ad libitum by the 

end of the study, but remained within 9 % that of the 

weight-matched controls through the end of the study, 

indicating that the weight-matching protocol was 

successfully employed. 

Feed Consumption, Clinical Findings, 

and Organ Weights 

Feed consumption by exposed males was similar to 

that by the controls during the study; females in the 

24,000 ppm group consumed less feed than the 

ad libitum-fed controls from week 38 through the end 

of the study (Table K1). There were no clinical findings related to butyl benzyl phthalate administration. 

The absolute and relative right kidney and liver 

weights of exposed males and females were greater 

than those of the weight-matched controls (Table Ila). 
34 Dietary Restriction, NTP TR 460 

TABLE 2 

Survival of Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

Ad Libitum- Weight-Matched 12,000 ppm 

Fed Control Control 

Male 

x~ 

Animals initially in study 

15-Month interim evaluation a 10 10 10 

Moribund 19 ' 15 26 

Natural deaths 3 1 2 

Animals surviving to study termination 28 33 22 

Other a 1 

Percent probability of survival at the end of study b 57 68 44 

Mean survival (days) e 625 648 632 

Survival analysis a P--0.520 

Survival analysis e P=0.036 

Ad Libitum- Weight-Matched 24,000 ppm 

Fed Control Control 

Female 

Animals initially in study 60 60 60 

15-Month interim evalnation a 10 10 10 

Moribund 23 7 .20 

Natural deaths . 

2 2 1 

Afiima!s surviving to study termination 25 41 29 

Percent probability of survival at the end of study 50 82 59 

Mean survival (days) 651 672 645 

Survival analysis P=0.759N 

Survival analysis P=0.011 

a Censored from analyses 

b Kaaplan-Meicr determinations " 

c Mean of all deaths (uncensored, censored, and terminal sacrifice) 

d The results of the life table pairwise comparisons (Cox, 1972) with the controls fed ad libitum are in the exposed group column. A lower 

mortality in the exposed group is indicated by N. 

¢ Result of life table pairwise comparison with the weight-matched controls 

-. 
D~e~ary ~esCHcfion, NT~ TR 4(6~ 3~ 

1.00 

~ 0.95 

<_> 0.90 

> 0.85 

~ 0.80 

o 0.75 

>" 0.70 

.e. 0.65 I~0 

< fill 0.60

o 0.55 

~" 0.50 

0.45 

.......... ::..-I ,,~,s I ......... ; .......... :::.--.....-.-~.--...-~.~.--.:.~ ...... 

~ .......... i...I ~=~t:=:=:='='o, I ......... ~ ~ ~ ~ ..... 

~...~..~..'.::0:::: .............. ~.~........ ~...:::::::~:~::::::::::~::::::::::~:::i ..... 0.40 

10 20 30 40 50 60 70 80 90 100 110 

WEEKS ON STUDY 

1.00 

i 0.95-

0.90 

~, 0.85 

~ 0.80 

~- 0,75 

~ 0.70 

¢~ 0.65 

~ 0.60 

0,55 

0.50 

i :: 

.......... ',J~ 

FEMALE RATS 

~°°?~ .......... iiiiii i .......... iiiiii ~ ........... iii iii ~ .......... i .......... i_.. L 

10 20 30 40 50 60 70 80 90 100 110 

WEEKS ON STUDY 

]FIGURE 3 

]Kapnan-Mder Surviva9 Curves for Mane and ~ema~e ~ats A~n~n~stered ~uty~ ~enzy~ Phtha~ate 

~n ]Feed for 2 'Years: Ad Libitum ]Needing and Wdght-I~Iatched Controls ]Protocons 
36 Dietary Restriction, NTP TR 460, 

450 ............ :..i ................ !.. ......... .=..~,..o.®..o.•.O-.o.oio..-. ......................... :~.... 

500 l ............ ~ i .. " ~.o •", ~ ~.~.~, ~ ~. o'~.~ ~ ~ ,~.,~ = ', 

400 "1 ...... ....,.-.,.i~....~..~-.-.i-.~..~..~.,~. ..... ~-.~..~. ............... i ............... ~........... ~.--i.~.-., . 

350 _1... ...... i ..... i.%~....'~ ........ i..'Li...~ .......... i .......... " ..... i .............. ..i.........: ...... ~, ..... 

300 

25O 

Q 

O 200 --...~... .................................................................................. : ....... i ..................... 

Z 

150 "--

I~ 

~" " i: ............................... i ~ ........................... i • 

MALE RATS 

Ad Libitum-Fed Control 

1" " 

,. 

. . . . [] Weight-Matched Control 

. ̧, 100 ~ ..... . .................. =...i ...................................... ~ t2.ooo~p~ " 

0 ~ 1,5 30 45 60 75 90 105 

WEEKSON sTuDY 

.. ,, 

500 ~ i 

450 ........ : ...... ~i..~.... I ........................ i........................ .....~ .......................................... 

400 ..... : .......... ~i. ...... . ......... .i ......... '................ ~i ...... - ......... "~;=. ............. 

z 350 ................ i ................ i........... ,...................... ~,.- ........... @..::.i ............ :::.: ..... 

I 300 

. 

................ 

~ , ~ 

i! .......... . ~.....:~: :~ . ................ ............ -.:-..e.o-o:~o.,o 

0~000 ~ . . . ....... -i ................ :: .... 

u.I ~ . ~ ~ o ooo:. :: i • :: " 

250 .......... : ~..... ::ii:T;i : ....... : .......... ...... ~•• :-:,...:,i . " ~ ~ i ~ : 

~ i .El " .... " 

o 200 .. ............ = ::, -, ~ -: z~.~E~e:: ~..~.~..~...~..~.~..l~..~.,~..[].-~.~.-.....~..~:...-..---,: ~ , ::. ...... :: : ...... ::..... 

Z ,,~¢~- ~ ~ ii' FEMALERATS "l'i 

150 ~'~-~ ....... i i ! i • AdLibitum-FedContrel |'":: ..... 

~" i :: :: :: B Weight-MatchedCont,ol / ! 

100 ~ ............................................................... !.. ~ 2,,ooooo~ |--.:: ..... 

0 15 30 45 60 75 90 105 

WEEKS ON STUDY 

.-

.. 

.. 

FIGURE 4 

Growth Curves for Male and Female Rats Administered Butyl Benzyl Phthalate 

in Feed for 2 Years: Ad Libitum Feeding and Weight-Matched ~Controls Protocols 
-- 

Dietary Restriction, NTP TR ,~60 37 

Pathology and Statistical Analyses. 

This section describes the statistically significant or 

biologically noteworthy changes in the incidences of 

mononuclear cell leukemia and of neoplasms and/or 

normeoplastic lesions of the pancreas, adrenal medulla, 

urinary bladder, preputial gland, and mammary gland. 

Summaries of the incidences of neoplasms and nonneoplastic lesions and statistical analyses of primary 

neoplasms that occurred with an incidence of at least 5 % 

in at least one animal group are presented in Appendix A 

for male rats and Appendix B for female rats. Pathologic descriptions of neoplasms and normeoplastic lesions 

occurring in rats in the butyl benzyl phthalate study are 

provided in NTP Technical Report 458 (NTP, 1997c). 

Pancreas: The incidence of adenoma of the acinus was 

significantly greater in exposed males than in the 

ad libitum-fed or weight-matched controls (Tables 3 and 

A2a). No adenomas occurred in the weight-matched 

controls; however, one male each in the weight-matched 

control and 12,000 ppm groups had an acinar cell 

carcinoma. The incidence of adenoma in exposed males 

TABLE 3 

exceeded the overall NTP historical control incidence of 

these neoplasms in untreated control male rats fed • 

ad libitum; no incidences of pancreatic carcinomas were 

recorded for historical control males. The incidence .of 

hyperplasia of the pancreatic acinus was greater in 

exposed males than in the .ad libitum-fed or weightmatched controls. 

Adrenal Medulla: Exposed males had significantly 

greater combined inci~dences of benign or malignant 

pheochromocytoma than those in the weight-matched 

controls (Tables 3 and A2a). The exposed group incidences of these neoplasms were similar to those in the 

controls fed ad libitum. The incidence of benign pheochromocytoma and the combined incidence of benign or 

malignant pheochromocytoma in exposed males were 

within the historical control, range for these neoplasms in 

untreated males fed ad libitum; the incidences of these 

neoplasms in the weight-matched controls fell below the 

historical control range for untreated males. The incidence of hyperplasia was greater in exposed males than 

in the ad libitum-fed or weight-matched controls 

(Table 3). 

k 

Incidences of Neoplasms and Nonneoplastic Lesions of the Pancreas and Adrenal Medulla 

in Male Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

Pancreas, Acinus a 

Focal Hyperplasia b 

Adenoma 

Overall rate c 

Adjusted rate d 

Terminal rate e 

First incidence (days) 

Logistic regression test. g 

Logistic regression test a 

Carcinoma 

Overall rate 

(continued) 

Ad LibitumFgd Control 

50 

4 

3/50 (6%) 

10.7% 

3/28 (11%) 

729 (T) 

,:. 

0/50(0%) 

Weight-Matched 

Control 

50 

2 

0/50 (0 %) 

0.0% 

0/33 (0%) f 

1/50(2%) 

12,000 ppm 

50 

12 

I0150 (20%) 

41.0% 

8/22 (36%) 

709 

P=0.016 

P<0.001 

1150 (2%) 
31; Dietary Restriction, NTP TR 460 

TABLE 3 ' 

Incidences of Neoplasms and Nonneoplastic Lesions of the Pancreas and Adrenal Medulla ~ 

in Male Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum ~'eeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 12,000 ppm 

Fed Control Control 

- Pancreas, Acinus (continue.d) 

Adcnoma or Carcinoma ~ 

Overall rate 3•50 (6%) 1/56 (2%) 11/50 (22%) 

Adjusted rate , 10.7 % 3.0% ,, 42.7% 

Terminal rate 3•28 (11%) 1/33 (3%) 8/22 (36%) 

First incidence (days) 729 (T) 730 (T) 674 

Logistic regression test P=0.014 

Logistic regression test P'<0.001 

Adrenal Medulla 50 50 50 

Hyperplasia 6 6 12 

Benign Pbeochromocytoma 

Overall rate .9/50 (18 %) 3•50 (6%) 8•50 (16%) 

Adjusted rate 28.2% 8.5% 25.4% 

Terminal rate 6/28 (21%) 2/33 (6%) 3/22 (14%) 

First incidence (days) - 607 678 639 

Logistic regression test P=0.475N 

Logistic regression test P=0.086 

Malignant Pheochromocytoma 

Overall rate 2/50 (4%) 1/50 (2%) 2•50 (4%) 

Benign or Malignant PheochromocytomaJ 

Overall rate 10/50 (20%) 3/50 (6%) 10/50 (20%) 

Adjusted rate 30.1% 8.5% 33.3% 

Terminal rate 6/28 (21%) 2/33 (6%) 5/22 (23%) 

First incidence (days) 607 678 639 

Logistic regression test P=0.573N 

Logistic'regression test P=0.028 

.. (T)Terminal sacrifice 

a Number of animals with tissue examined microscopically 

b Number of animals with lesion 

¢ Number of animals with neoplasms per number of animals with tissue examined microscopically 

d Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

e Observed incidence in animals surviving until the end of the study 

f Not applicable; no neoplasms in animal group 

g In the exposed group column are the P values corresponding to the pairwise comparisons between the ad libitum-fed controls and the 

exposed group. The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the 

exposed group is indicated by N. . 

h Result of pairwise comparison between the weight-matched controls and the exposed group 

Historical incidence for 2-year NTP feed studies with untreated control groups fed ad libitum (mean ± standard deviation): 19/1,191 

(1.6% =l= 2.4%); range, 0%-10%. 

J Historical incidence: 398/1,182 (33.7% ± 10.4%); range, 12%-63%. 

i 
Dietary Restriction, NTP TR 460 39 

Urinary Bladder: Exposed females had a greater Mononuclear Cell Leukemia: The incidences of 

incidence of hyperplasia of the urinary bladder transi- mononuclear cell leukemia in exposed males and 

tional epithelium (10/50) than did the ad libitum-fed females were greater than those in the weight-mat6hed 

controls (4/50) or the weight-matched controls (0/50) controls (Tables 4, A2a, and B2a) but similar to the 

(Table B3a). incidences in the controls fed ad libitum anal within 

the historical control ranges for leukemia (all types) in 

untrreated rats. 

TABLE 4 

]Incidences of Mononuclear Cela Leukemia in Rats in the Dietary Restriction Study 

of ~utyl ll~enzyl Phthalate: Ad Libitum Peedlng and Wefight-Matched Controls IProtoco~s 

Ad Libitum- Weight-Matched 12,0~ ppm 

Fed Control Control 

Male 

Mononuclear Celi Leukemia a 

Overall rate b 31/50 (62%) 15/50 (30%) 30/50 (60%) 

Adjusted rate c 71.8% 34.9% 65.6% 

Terminal rate d 16/28 (57%) 6/33 (18%) 7/22 (32%) 

First incidence (days) 479 422 180 

Life table test e P=0.478 

Life table test f P=0.002 

Ad Libitum- Weight-Matched 24,000 ppm 

Fed Control Control 

Female 

Mononuclear Cell Leukemia g ~ 

Overall rate 21/50 (42%) .13/50 (26%) 19/50 (38%) 

Adjusted rate 51.7 % 28.6% 46.1% 

Terminal rate 7/25 (28%) 9/41 (22%) 8/29 (28%) 

First incidence (days) 368 551 452 

Life table test P=0.398N 

Life table test P=0.034 • 

Historical incidence for 2-year NTP feed studies with untreated control groups fed ad libitum(mean =l: standard deviation): 562/1,203 

(46.7% 5: 10.7%); range, 18%-62%. Includes lymphocytic, monocytic, mononuclear cell, and undifferentiated leukemias. 

b Number of animals with neoplasms per number of animals necropsied 

c Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

d Observed incidence in animals surviving until the end of the study ' 

• In the exposed group column are the P values corresponding to the pairwise comparisons between the ad libitura-fedcontrols and the 

exposed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of 

death. A lower incidence in the exposed group is indicated by N. 

f Result of pairwise comparison between the weight-matched controls and the exposed group 

g Historical incidence: 322/1,202 (26.8% 5: 9.0%); range, 14%-52%. Includes lymphocytic, monocytic, mononuclear cell, and 

undifferentiated leukemias. 

i 
-- 

40 

Preputial Gland: The combined incidence of preputial gland adenomas and carcinomas was significantly less in exposed males than in the controls fed 

ad libitum and the weight-matched controls (Tables 5 

and A2~)i these neoplasms occurred in five controls 

fed ad libitum and in six weight-matched controls, but 

in no exposed males. 

Mammary Gland: The incidence of mammary gland 

fibroadenomas and the combined incidence of fibroadenoma, adenoma, or carcinoma in exposed females 

were significantly less than those in the controls fed 

~;Y . 

• 

.TABI~E 5 

Dietary Restriction, NTP TR 460 

ad libitum; the incidences of these neoplasms in 

exposed females were similar to those in weightmatched control ' females (Tables 5 and B2a). Two 

ad libitum-fed control females with fibroadenomas 

also had • 

adenomas; .another control female with 

fibroadenoma and one additional control female also 

had carcinomas. Additionally, exposed females had 

a lower incidence Of mammary gland hyperplasia than 

that of the controls fed ad libitum; the incidence in 

exposed females was greater than that in the, weightmatched controls (Table 5). 

/-/ Incidences of Selected Neoplasms and Nonneoplastic Lesions in Rats 

in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

Male 

Preputial Gland 

Adenoma 

Overall rate a 

Adjusted rate b 

Terminal rate c 

First incidence (days) 

Logistic regression test e 

Logistic regression test f 

Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression,test 

Adenoma or Carcinoma g 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

(co~ti-.,=d) 

A~ LibitumFed Control 

4/50 (8%) 

12.8% 

2/28 (7%) 

644 

1/50 (2%) 

2.7% 

0/28 (0%) 

614 

5/50(10%) 

15.1% 

2/28 (7%) 

614 

Weight-Matched 

Control 

3/50 (6%) 

8,5% 

2/33 (6%) 

678 

3/50 (6%) 

8.2% 

2/33' (6%) 

607 

6/50 (12%) 

16.4% 

4/33 (12%) 

607 

12,000 ppm 

O/50 (O%) 

0.0% 

0/22 (0%) _d 

P=0.059N . . . 

P=0.136N 

0/50 (0%) 

0.0% 

0/22'(0%) 

P=0.504N 

P=0.124N 

0/50 (0 %) 

0.0% 

0/22 (0%) 

P=0.032N 

P=0.020N 
Dietary Restriction, N T P T R 460 41 

TABLE 5 

Incidences of SdecCed Neoplasms and Nonneoplmfic Lesions ~n Rats 

•in the Dietary Restriction Study of Butyl Benzyi Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) . 

Ad Libitum- Weight-Matched 24,000 ppm 

Fed Control Control 

~¢emale 

Mammary Gla.nd h 50 50 50 

Hyperplasia t 30 9 17 

Fibroadenoma 

Overall rate 28/50 (56%) 7/50 (14%) 11/50 (22%) 

Adjusted rate 71.0 % 16.5 % 28.9% 

Terminal rate 14/25 (56%) 6/41 (15%) 5/29 (17%) 

First incidence (days) 587 722 487 

Logistic regression test P<0.001N 

Logistic regression test P=0.225 

Adenoma 

Overall rate 2/50 (4%) 0/50 (0%) 0/50 (0%) 

Carcinoma 

Overall rate 2/50 (4%) 0/50 (0%) 0/50 (0 %) 

Fibroadenoma, Adenoma, or Carcinoma j 

Overall rate 29/50 (58%) 7/50 (14%) 11/50 (22%) 

Adjusted rate 71.8% 16.5% 28.9% 

Terminal rate 14/25 (56%) 6/41 (15%) 5/29 (17%) 

First incidence (days) 587 722 487 

Logistic regression test P<0.001N 

Logistic regression test P=0.225 

a Number of animals with neoplasms per number of animals examined. Denominator is number of animals examined microscopically for 

preputial gland and number of animals necropsied for mammary gland. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

e Observed incidence in animals surviving until the end of the study 

d Not applicable; no neoplasms in animal group 

e In the exposed group column are the P values corresponding to the pairwise comparisons between the ad libitum-fed controls and the 

exposed group. The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the 

exposed group is indicated by N. 

f Result of pairwise comparison between the weight-matched controls and the exposed group 

g Historical incidence for 2-year NTP feed studies with Untreated control groups fed ad libitura (mean 5: standard deviation): 84/1,164 

(7.2% 5: 5.6%); range, 0%-24%. 

h Number of animals necropsied 

i Number of animals with lesion 

J Historical incidence: 507/1,202 (42.2% 5: 13.5%); range, 8%-64%. 
42 Dietary Restriction, NTP TR 460 

2-YEAR AND LONG-TERM RESTRICTED FEED PROTOCOLS • 

Survival 

Estimates of 2-year and long-term survival probabil. 

ities for male and female rats fed restricted diets for 

up to 32 months are shown in Table 6 and in the 

Kaplan,Meier survival curves in Figure 5. The 

survival rates of exposed males and females were 

similar to those of the controls at 2 years and at the 

end of the study. Survival was reduced to 20% 

during week 129 for males and week 140 for females. • , 

: 

Body Weights 

Mean body. weights are given in Figure 6 and in 

Table J2. At 1 year, the mean body weights of 

exposed male rats in each restricted feed protocol 

were 95 % those of the respective controls; the mean 

body weights of exposed females in the 2-year and 

long-term protocols were 93% and 91% those of the 

controls, respectively, at 1 year. From 1 year through 

the end of the study, the. mean body weights of 

exposed males remained within 10% of the controls; 

exposed females in the 2-year restricted feed protocol 

weighed 23% less than the controls at the end of 

exposure, and exposed females in the long-term 

restricted feed protocol weighed 29% less than the 

controls at the end of exposure. 

Feed Consumption, Clinical Findings, 

and Organ Weights 

Feed consumption data are shown in Table K2. There 

were no clinical findings related to butyl benzyl 

phthalate administration. At the 15-month interim 

evaluation, exposed males had significantly greater 

absolute and relative right kidney weights than those 

of the controls (Table lib). 
c 

Dietary Restriction, NTP TR 460 43 

TAnL~ 6 • 

Survival of ~ats ~n the Dietary ]Restriction Study of ~uty~ ]~enzy~ Phtha~ate: 

~-~ear and Long-Ter~n Restricted Feed ~'rotoco~s 

~Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

Male 

Animals initially in study 60 60 50 50 

15-Month interim evaluation a 10 10 0 0 

Accidental death a 1 0 0 0 

Moribund 14 17 37 29 

Natural deaths 1 2 3 8 

Animals surviving to study termination ~-~4 31 10 13 

Percent probability of survival at the end of study b 70 62 20 26 

Mean survival (days) c 656 655 763 730 

Survival analysis d P=0.537 P=0.731N 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Female 

Animals initially in study 60 60 50 50 

15-Month interim evaluation a 10 10 0 0 

Moribund 11 10 34 38 

Natural deaths 4 1 6 1 

Animals surviving to study termination. 35 39 10 11 

Percent probability of survival at the end of study 71 78 20 22 

Mean survival (days) 654 666 803 845 

Survival analysis P=0.473N P=0.522N 

a Censored from analyses 

b Kaplan-Meier determinations 

Mean of all deaths (uncensored, censored, and terminal sacrifice) 

d The result of the life table pairwise comparison (Cox, 1972) with the contxols is in the exposed group column. A lower mortality in the 

exposed group is indicated by N. 
44 Dietary Restriction, NTP TR460 

1.0 ~ ~ = : ~-,:-"~! ...................................... 

_ 

0.5 ....... ~ MALE RATS ...... :: ......... :, "':' ........ 

'~ ,~sT.,oT~o~o i i~:.i.~_i. 

0.4 ....... ~ ~ ~.~ • ..... ) ........ ~ ~ ~ - ~ .... 

~ 0.3 t~°~°~°) ~ ~ ~ ~ :: ~ I ...... I ~ 12,000ppm(2yr) ...... :: ......... ~ ~ ~ ' ~ - ~ 

0.2 ........................ ~ ~ ...... ; ; ~ 

0.1 ~ 

I0 20 30 40-50 60 70 80 90 100 110 1201~0 

WEEKS ON STUDY 

< 

1.0 i~ *~;~~ ......::i::i::i:::::',::::i:i• .~ 0.9 

>_ 

> 0.8-

D

¢n 0.7 

Lt. 

o 0.6 >-

I - FEMAt:ERATS I:: :: i i ~ 0.5 -:"I RESTR~OTEO~EO ~I~....... : : - - ~ -

<= 0.4 I + o ~p~ (~ ~,) ~:~ ~ ~ ~ ........ I ...... l ~ 0 ppm (30 too) l~ ~ ~ :: ~ ~ ~ ~ l O I ~ 24.000 ppm (2 yr) i ~ ~ ~ ....... ~........ i =. 0.3 ...... i~,,.ooo,,~(,o.o, ~ ....... ::--.~~ .......~ ....... ~. ....... i ....... ~ ........ ~........ ~........ ~ ....... ~ . . . ~ ..... 2 ...... ~.. 0.2 : ~ - ~ 

0.1. I I 

~0 20 aO 40 50 60 70 80 90 ~00~0~20~a0~40 

WEEKS ON STUDY 

FIGURE 5 

Kaplan-Meier Survival Curves for Male and Female Rats Administered Butyl Benzyl Phthalate 

in Feed for up to 32 Months: 2-Year and Long-Term Restricted Feed Protocols 
D~e~ry Restriction, NTP TR 460 45 

~ 

z 

~ 

~ 

o 

Z 

< 

~ 

~< 

O 

~. 

o 

~ 

~ 

~ 

~ 

450 

400 

350 

300 

250 

200 

150 

100 

50 

450 

400 

350 

300 

250 

200 

150 

1 O0 

50 

_~........................... ~ ....... .i., ......... o.o~.~.~! ...... o..i ............. ! ...................... 

/ i ~j~~.~i~d~_~;oo i 

d ............. iBe~~~~ ~~ ...... i ............. i# ........ ~~~; ..... .:_,;:.it ........ I 

I i_O ! i i : : ~V~DD I .......... ' ............. ' ............. ' ............. '............. ' ............. ......... 

MALE RATS I I ...................................... . ............................ 

- ~ ........... i ........................................................... 0 ~12yO 

[] 0 ppm (30 too) 

~ 12.000 ppm (2 yr) 

...................................... : ................................ ~ 12,000 ppm (30 too) 

! ~ ~ 

I I I 

0 15 30 45 ~60 75 90 105 120 

WEEKS ON STUDY 

~ 

......................................................... .:;: ............................................................... 

' ............ ............ ............. .............i ............. ............. ............. ............. ........ :: i 

................... ~ ...... ~ i ............. ............. ! ............. i ....... ~~~~~~~~~~o 

.............i............. i ............. i ............. ! .......... Oi.O. ~ ...... ~ ............. i ............. ~........ ~ ~ ~ ~g ~ :: ~ ~ : ~ ~v~ ~ : ~ ~ ~ ? ............. ~--.-:.~-.~._-~.~.~-~-~.~_-~.-~ . . . . . . 

..~~ ~ ~ ~ ~ ~.~T~ ~ 

~ .............. ~ ............. ~............. ~............. ~ .... ,EST.,OTE~FEE~ I ..... 

E ......... L ............ ~............. :: ............. :~ ............. L.. o 0..~ (= ~r) I ..... ~ ~ ~ ~ ~ ~ Oppm(32mo) l ~ ~ ~ ~ ~ ~ 24,000 ppm (2 yr) I 

............. :.............. : ............. ~............. , ............. :..- ~ ~.OOOpp~mo~ I ..... 

0 15 30 45 60 75 90 105 120 

WEEKS ON STUDY 

i 

i t~.-[ 

, . .i: 

FIGURE 6 

Growth Curves for Male and Female Rats Administered Butyl Benzyl Phthaiate 

in Feed for up to 32 Months: 2-Year and Long-Term Restricted Feed Protocols 
46 

Pathology and Statistical Analyses 

This section describes the statistically significant or 

biologically noteworthy changes in the incidences of 

neoplasms and nonneoplastic lesions of the urinary 

bladder, clitoral gland, mammary gland, pituitary 

gland, and skin. Summaries of the incidences of 

neoplasms and nonneoplastic lesions and statistical 

analyses of primary neoplasms that occurred with an 

incidence of at least 5 % in at least one animal group 

are presented in Appendix A for male rats and Appendix B for female rats. 

Urinary Bladder: At 2 years and at 32 months, 

exposed females had significantly greater incidences 

of hyperplasia of the urinary bladder transitional 

epithelium than did the controls (Tables 7 and B3b). 

At 32 ~ months, exposed ~ female rats had a slightly 

greater incidence of papilloma or carcinoma (combined) of the urinary bladder transitional epithelium 

than the controls (Tables 7 and B2b); two exposed 

females had papillomas, and four exposed females had 

carcinomas. One control female had a papilloma at 

32 months. Papillomas also occurred in two exposed 

females at 2 years. One exposed male had a papilloma at 2 years; at 30 months~ one exposed male had 

Dietary Restriction, NTP TR 460 " 

a papilloma and a carcinoma (Table 7). No control 

males had urinary bladder neoplasms. ' 

Clitoral Gland: The incidence of clitoral gland 

carcinoma and the combined incidence of clitoral 

gland adenoma or carcinoma in exposed females were 

less than those in the controls at 32 months (Tables 8 

and B2b). 

Mammary Gland: At 2 years and at 32 months, the 

incidences of fibroadenoma and the combined incidences of fibroadenoma or carcinoma in exposed 

females were significantly less than those in the 

controls (Tables 8 and ~B2b). 

Pituitary Gland." The incidences of adenoma of the 

pars distalis in exposed females were less than those 

in the controls at 2 years and at 32 months (Tables 8 

and B2b). Additionally, one control female had a 

Carcinoma of the pars distalis at 32 months. • 

Skin: Four control females had fibromas of the 

subcutaneous tissue at 32 months; no fibromas were 

observed in exposed females (Tables 8 and B2b). 
i 

J 

Dietary Restriction, NTP TR ,~6~ 

TABLE 7 

~c~dences o~' Heop~m~ns and Nonneop~mt~c Lesions 

o~ the Ur~aary ~nadder '~ransitiona~ EpRhe~iu~n in Rats in the Dietary Restriction. Study 

o~ ~uty~ ~e~3'~ Phtha~ate: 2-Year and Long-Te~'rn Restricted ~'eed ]Protocols 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

~ale 

Number Examined Microscopically 50 50 50 50 

Hyperplasia a 1 2 0 1 

Papilloma 

Overall rate b 0/50 (0%) 1/50 (2%) 0/50 (0%) 1/50 (2%) 

Carcinoma ~ 

Overall rate 0/50 (0%) 0/50 (0%) 0•50 (0%) 1/50 (2%) 

2-Year Restricted Feed 32-Month Restricted Feed 

Female 

; 

0 ppm 2,~,000 ppm 0 ppm 2~,000 ppm 

Number Examined Microscopically 50 50 50 

Hyporplasia 0 14"* 0 16"* 

Papilloma 

Overall rate 0/50 (0%) 2/50 (4%) 1/49 (2%) 2/50 (4%) 

Carcinoma 

Overall rate 0/50 (0%) 0•50 (0%) 0/49 (0%) 4/50 (8%) 

Adjusted rate c 0.0% 29.O% 

Terminal rate d 0/10 (0%) 3/11 (27%) 

First incidence (days) • • - 719

Logistic regression test f P=0.079 

Papilloma or Carcinoma 

Overall rate 0/50 (0%) 2/50 (4%) I149 (2%) 6/50 02%) 

Adjusted rate 2.6% 41.0% 

Terminal rate 0/10 (0%) 4/11 (36%) 

First incidence (days) 694 ,, 719 

Logistic regression test P=0.077 

** Significantly different (P~0.01) from the control group by the Fisher exact test 

a Number of animals with lesion 

b Number of animals with neoplasms per number of animals with urinary bladder examined microscopically 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

d Observed incidence in animals surviving until the end of the study 

e Not applicable; no neoplasms in animal group 

f In the exposed group columns are the P values corresponding to the pairwise comparisons between the controls and that exposed group. 

The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. 

47 

c 
Dietary Restriction, NTP TR 460 

TABLE 8 

Incidences of Selected Neoplasms in Female Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and Long-Term Restricted Feed Protocols~ 

Clitoral Gland 

Adeno_ma 

Overall rate a 

Adjusted rate b 

Terminal rate c 

First incidence (days) 

Logistic regressio~ test d 

Carcinoma 

O~erall rate ~ 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Mammary Gland 

Fibroadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Fibroadenoma or Carcinoma 

Overall. rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

(continued) . - 

2-Year Restricted Feed 

0 ppm 24,000 ppm 

3•50 (6%) 3•50 (6%) 

16.4% 8.8% 

2•35 (6%) 3/39 (8%) 

677 731 (T) 

P=0.636N 

0/50 (0%) 0/50 (0%) 

3/50 (6%) 3/50 (6%) 

16.4% 8.8% 

2/35 (6%) 3/39 (8%) 

677 731 (T) 

P=0.636N 

12/50 (24%) 2/50 (4%) 

44.9% 5.0% 

12/35 (34%) 1/39 (3%) 

730 (T) 600 

P=0.003N 

1/50 (2 %) 0/50 (0 %) 

13/50 (26%) 2/50 (4%) 

46.4% 5.0% ' 

12/35 (34%) 1/39 (3%) 

712 600 

• P=0.002N 

32-Month Restricted Feed 

0 ppm 24,000 ppm 

5/50 (10%) 

16.2% 

0/10 (0%) 

649 

5/50 (10%) 

38.5% 

3/10 (30%) 

829 

10/50 (20%) 

48.5 % 

3/10 (30%) 

649 

21/50 (42%) 

78.9% 

5/10 (50%) 

687 

4/50 (8%) 

14.8% 

o/10 (0%) 

694 

24/50 (48 %) 

81'.5% 

5/10 (50%) 

687 ,. 

2/49 (4%) 

11.7% 

1/11 (9%) 

779 

0.253N 

0/49 (0%) 

o.o% 

0/11 (O%) _~ 

0.021N 

2/49 (4%) 

11.7% 

1/11 (9%) 

779 

P=0.012N 

5/50 (10%) 

26.1% 

2/11 (18%) 

796 

p<0.001N 

1/50 (2%) . 

9.1% 

1/11 (9%) 

977 (T) 

P=0.173N 

6/50 (12%) 

34.4% 

3/11 (27%) 

796 

P<0.001N 
D~et~ry ~estricfion~ NT~ T~ ,~60 49 

TABLE 8 

Incidences o~ Selected Neoplmms in ~enaa~e ~ats ~n the Dietary Restriction Stagy 

of ~uty~ ~enzy~ ]?hthalate: 2-Year and Long-Term ~eestr~cted ~eed ]l~rotoco~ (continued) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,~00 ppm 

Pituitary Gland (Pars Distalis) 

Adenoma 

Overall rate 15/50 (30 % ) 6/49 (12 % ) 25/50 (50 % ) 16/50 (32 % ) 

Adjusted rate 45.8% 14.9% 85.4% 81.8% 

Terminal rate 12/35 (34%) 4/39 (10%) 7/10 (70%) 8/11 (73%) 

First incidence (days) 564 600 478 731 

Logistic regression test P=0.023N P=0.018N 

Carcinoma 

Overall rate 0/50 (0%) 0/50 (0%) 1/50 (2%) 0/50 (0%) 

Skin (Subcutaneous Tissue) 

Fibroma 

Overall rate 1/50 (2%) 0/50 (0%) 4/50 (8%) 0/50 (0%) 

Adjusted rate 28.0% 0.0% 

Terminal rate 2/10 (20%) 0/11 (0%) 

First incidence (days) 687 -

Logistic regression test P=0.048N 

(T)Terminal sacrifice 

a Number of animals with neoplasms per number of animals examined. Denominator is number of animals examined microscopically for 

clitoral gland and pituitary gland and number of animals necropsied for mammary gland and skin. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

e Observed incidence in animals surviving until the end of the study 

d In the expose d group columns are the P values corresponding to the pairwise comparisons between the controls and that exposed group. 

The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence inan exposed group is 

indicated by N. 

e Not applicable; no neoplasms in animal group 
~;0 Dietary Restriction, NTP TR 460 

t-BUTYLHYDROQUINONE FEED STUDY 

AD LIBITUM FEEDING AND WEIGHT-MATCHED CONTROLS PROTOCOLS ~ ,, 

Survival 

Estimates of 30-month survival probabilities for male 

and female rats fed ad libitum and the. weight-matched 

control rats are shown in Table 9 and in the KaplanMeier survival curves in Figure 7. The survival rates 

of males and females administered 5,000 ppm were 

greater than those of the controls fed ad libitum and 

similar to those of the weight-matched controls. 

Survival was reduced to 20% during week 123 for 

exposed males and week 129 for exposed females. 

Body Weights 

Mean body weights are given in Figure 8 and in 

Table J3. The mean body weight of males in the 

5,000 ppm group was 92% that of the controls fed 

ad libitum and 103% that of the weight-matched 

controls at 1 year; the mean body weight of exposed 

males remained within 8 % that of the controls fed 

ad libitum and within 5% of the weight-matched 

controls from week 53 until the end of the study. The 

mean body weight of females in the 5,000 ppm group 

was 90% that of the controls fed ad libitum and 

- ̧. 

104% that of the weight-matched controls at 1 year. 

From-week 53 until the end of the study, exposed 

females weighed 8% to 15% less than the controls fed 

ad libitum and 2% to 11% more than the weightmatched controls. 

Feed Consumption, Clinical Findings, 

and Organ Weights 

Feed consumption by exposed animals was similar to 

that by the controls fed ad libitum (Table K3). 

Clinical findings included skin discoloration in 

exposed males and females and urine stain on the fur 

of exposed females. 

At the 3-month interim evaluation, exposed females 

had a lower absolute right kidney weight than that of 

the controls fed ad libitum (Table I2a). The absolute 

right kidney weight of exposed males and the absolute 

and relative liver weights of exposed males and 

females were greater than those of the weight-matched 

controls. 
c 

D~e~ry Restriction, NTP T~ ~60 5~ 

TABLE 9 

Survival o~ ~a~ ~n the Dietary Restriction Study of t-~uty~hydroquinone: 

Ad Libitum ~eed~ng and Weight-Matched Controls ~rotoco~s 

Ad Libitum- Weight-Matched 5,000 ppm 

Fed Control Control 

Made 

Animals initially in study 70 70 70 

3-Month interim evaluation a I0 10 10 

Moribund 48 41 42 

Natural deaths 4 7 4 

Animals surviving to study termination 8 12 14 

Percent probability of survival at the end of study b 13 20 23 

Mean survival (days) ¢ 621 657 629 

Survival analysis d P=0.361N 

Survival analysis e P=0.619 

Female 

Animals initially in study 70 70 70 

3-Month interim evalnation a 10 10 10 

Moribund 40 31 36 

Natural deaths 10 7 7 

Animals surviving to study termination 10 22 17 

Percent probability of survival at the end of study 17 37 28 

Mean survival (days) 636 697 693 

Survival analysis P = 0.030N 

Survival analysis P=0.481 

a Censored from analyses 

b Kaplan-Meier determinations 

Mean of all deaths (uncensored, censored, and terminal sacrifice) 

d The results of the life table pairwise comparisons (Cox, 1972) with the controls fed ad libitumare in the exposed group column. A lower 

mortality in the exposed group is indicated by N. 

e Result of life table palrwise comparison with the weight-matched controls 
_ _ 

52 Dietary Restriction, NTP TR 460 

-_ 

-,_ 

1.0 

0.9 

0.8 

0.7 

0.6 O 

0.5 

0.4 < 

0.3 O 

0.2 

0.1 

0.0 

1.o 

0.9 

o.o 

°.~ 

0.6 

o.s 

0.4 

• o.a 

u.z~ 

0~.-

: ,: : : , ., : 

, • ! : 

r : ~ : ~''''" ..... ::~ :'~ ..... ......... ~.................. ~ ........ 

.. 

................. ~................. ~..~ ............... ~ ................. i ................. .~ ...... '. .~..: ...... 

I - - - ~ , 

/ I --e- A d~ub..~ed c~t,o~. ~ ~ . ~ . .'~ 

~"" ......... L ~ W~ht-Mat~ed C~trot ~ ......... ~ .... ~'--'~ ...... 

~ ............ ~ ~:.0~,,~ ;.. ........ ~..~.~.~ .... 

_ ~ ~ ,,~ . ;., . ~ ,. ~ , . ,,; 

20 40 60 80 • 100 120, 

WEEKS ON STUDY " • • .. 

! ": 

. 

:, • 

; - : ; 

:: " ~ :i ........... : ..... ".................. ::i" 

........... i..... :.o. .............. i.................. : i ..... i--~ ............. i ........ 

- ............... i., ............... 

~~ ....... ~i ......... ~' ..... i ............ ::i:i ....... . ......... i ...... ~~.~-~;~"i ...... ~ ............... :.~: ........ : ....... ; ........... '.--.::~,.-.,: ........... ~................. ~ .... i ...... 

J ................. ~ ................. :~ ................. ~-,.-~ ....... :-.:~.:-.,....... ~..,~ ............ ............ I Fe~ 

~ " ~~,.~-~.~*~ I ~ ~ • : 1~ " ~ ............ I~ ~,.~=~.-.~:::-~ 

/ / ~ 5,0~ppm ~ ~ ~ - : ........... .:t ;....;. : ......... ; ...... ~.--.; ........... :-",. ..... : ...... ~ ..... 

~, , ~, :~ ~,~ " ~, 

,. , 

20~' 40 60 " ~ 80. 100 120 

WEEKS ON STUDY 

FIGURE 7 , ., • 

Kaplan-Meier Survival Curves for Male and Female Rats Admini~ered t-Butylhydroquinone 

in Feed for up to 30 Months: Ad Libitum Feeding and Weight-Matched .Controls,Protocols 
Die~ry Restricfionl NT~ TR 4~0 53 

500 

_ i _ oo~ooO::ooo i i 

450 -- ............. ~............ ~ ........ O-U. O- ........ ~,- ....... :---i .......... .d.Q.o. o ............ ....:, ........ 

< ~ ~ o4OO.~~.~,°0o~~ao~ I 

400 .............. ~.--o ..... 5z~-8-a.~.a ....... ! ............. ~ ............ ~............. ~..... -~.a-~ ...... I 

~.~ ~ ~ ....j: ............ ~ ............ ~............. ~ ............. ~ ........I 

350 .......... ~~6 a .......................... ~ ' { I 

w 

>-

o 

Z 

g 

i i ~ 

0 :15 30 45 60 75 90 105 120 

WEEKS ON STUDY 

500 : . , . 

450 ......................................... ~-............................................................................ : 

: 

< 400 ............................... "......... :{ ........................... !.................................................. 

; • 

350 ............. !i ............ ": ...... : ...... i............. : ............. ~ ............ :'~'oo'o'~"6"o'b" ...... 

I :: i :: :: o::O ° ° °. ." ~. ~ 300 -I ........... "i" ......... i............. i......... "'J"~ 'o° '"~"'"7"Z~ z~~ ~~'~~ ~ia ...... t.IJ | ~ i i i~o~~i~e. ~D i •---- !- ..... ":'............. 6 ......... ~a'a-~ "~~ ................................. ! ........ 

~so-1 • og~g~,~ ! ~ . i ~. o ~o0 -] -~;~1.~- '~. ~i...... ~........... I " ' ~~ms l ...... i " : .... ~ ........ Z < 150 .-t.2,4i~I~"-i-, .......... i ..... : ....... i ....... • ..... i ........... I o ~ou~,m-~aco~o~ ~.i ........ I..LI 

/~' :: ' ' ' I X N,~'2 '~°'°°°'*°' [~ 

.~oo ~-..~ ...... , ............ i...... -i ....... : ..... ' ........... ,, '~" , ,i ........ 

0 . 15 aO., 45 60 "75 90 105 120 

• 

• . WEEKS ON STUDY ~-

]~'IGURE ~ 

Growth Curves for IV~ale an¢~ ~ema~e ~a~s Achnlnis~ere~ t-~utylhydroquinone 

~n ~eed for up to 30 Months: Ad Libitum ~eed~ng and.Weight4V~atched Controls Protocols 

. . 

G~ey Draft NOT YOR D~ST~BUT~ON O~ ATT~UT~ON 
54 

Pathology and Statistical Analyses 

This section describes the statistically significant or 

biologically noteworthy changes in the incidences of 

neoplasms of the preputial gland, clitoral gland, 

pancreas, pituitary gland~ and mammary gland. 

Summaries of the incidences of neoplasms and nonneoplastic lesions and statistical analyses of primary 

neoplasms that occurred with an incidence of at least 

5 % in at least one animal group are presented in 

Appendix C for male rats and Appendix D for female 

rats. Pathologic descriptions of neoplasms and 

nonneoplastic lesions occurring in rats in the t-butylhydroquinone study are provided in NTP Technical 

Report 459 (NTP, 1997d). 

TABLE 10 

Dietary Restriction, NTP TR 460 

Preputial Gland: Males exposed to t-butylhydroquinone had significantly greater incidences of preputial gland carcinoma and adenoma or carcinoma 

(combined) than did the weight-matched controls 

(Tables 10 and C2a); there were no significant differences in the incidences of these neoplasms between 

exposed males and controls fed ad libitum. 

Clitoral Gland: Exposed females had significantly 

greater incidences of clitoral gland carcinoma and 

adenoma or carcinoma (combined) than did the 

weight-matched controls (Tables 10 and D2a); the 

incidences of these neoplasms in exposed females 

were similar to those in the controls fed ad libitum. 

Incidences of Preputial and Clitoral Gland Neoplasms in Rats 

in the Dietary Restriction Study of t-Butyihydroquinone: 

• Ad Libitum Feeding and Weight-Matched Controls Protocols 

Male 

Preputial Gland 

Adenoma 

Overall rate a " ~ .

Adjusted ra.te b 

Terminal rate c 

" - First incidence (days) 

Logistic regression test e 

Logistic regression test f 

Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

~ -Logistic regression test 

Adenoma or ,Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

(continued) 

Ad ILibituraFed Control 

5/6O (8%) 

31.1% 

2/8 (25%) 

528 • 

2/60 (3%) 

3.6% 

0/8 (0%) 

381 

7/60 (12%) 

33.6% 

2/8 (25%) 

381 

Weight-Matched 

Control 

0/6O (0%) 

0.0% 

0/12 (0%) _d 

0/60 (0%) 

0.0% 

0/12 (0%) 

0/6O (0%) 

0.0% 

0/12 (0%) 

5,000 ppm 

3/60 (5%) 

18.4% 

2114 04%) 

786 

P=0.312N 

P=0.084 

5/60(8%) 

18.4% 

1/14 (7%) 

520 

P=0.200 

P=0.040 

8/60 (13%) 

34.2% 

3/14 (21%) 

520 

P=0.556 

P=0.004 
Die~ry ]Restriction, NTIP T~ ~ 

TA~L~ ~0 

~Incidences of ~'reputial and Clitoral Gland Neoplasms in ~a~s 

ha the ~ietary ]Restriction Study of t-~uty~hydroquinone: 

Ad Libigura ]Feeding and Weight-Matched Controls ]Protocols (continued) 

Ad Libitum- Weight-Matched 5,000 ppm 

Fed Control Control 

Fenmle 

Clitoral Gland 

Adenoma 

Overall rate 6/58 (10%) 4/59 (7%) 7/60 (12%) 

Adjusted rate 38.3% 14.5% 27.0% 

Terminal rate 3/10 (30%) 2/22 (9%) ' 2/17 (12%) 

First incidence (days) 579 817 628 

Logistic regression test P = 0.513N 

Logistic regression test P=0.247 

Carcinoma 

Overall rate 6/58 (10%) 2/59 (3%) 8/60(13%) 

Adjusted rate 43.2% 4.0% 29.0% 

Terminal rate 4/10 (40%) 0/22 (0%) 2/17 (12%) 

First incidence (days) 649 648 750 

Logistic regression test P=0.579N 

Logistic regression test P=0.050 

Adenoma or Carcinoma 

Overall rate 12/58 (21%) 6/59 (10%) 14/60 (23%) 

Adjusted rate 74.9% 17.9% 47.4% 

Terminal rate 7/10 (70%) 2/22 (9%) 4/17 (24%) 

First incidence (days) 579 648 628 

Logistic regression test P=0.402N 

Logistic regression test P=0.040 

a Number of animals with neoplasms per number of animals with tissue examined microscopically 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

c Observed incidence in animals surviving until the end of the study 

d Not applicable; no neoplasms in animal group 

© In the exposed group column are the P values corresponding to the pairwise comparisons between the ad libitum-fed controls and the 

exposed group. The logistic regression test regardsneoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the 

exposed group is indicated by N. 

f Result of pairwise comparison between the weight-matched controls and the exposed group 
56 Dietary Restriction, NTP TR 460 

Pancreas: Three control males fed ad libitum had 

adenomas of the pancreaticacinus;:this-ne0plasm did 

not occur in the .exposed, males orin the weigh t - 

matched controls (Tables 11 and C2a). 

Pituitary Gland, Pars Distalis: The incidences of 

adenoma and adenoma or carcinoma (combined) were 

significantly less in exposed males than in the controls 

fed ad libitum or the weight-matched controls 

(Tables 11 and C2a). 

~. 

Mammary Gland: Exposed females had significantly 

lower incidences of fibroadenoma; fibroaden6ma or 

adenoma (combined); and fibroadenoma, adenoma, or 

carcinoma (combined) than did the confrols fed 

ad libitum (Tables 11 and D2a). Additionally, there 

were fewer mammary gland carcinomas in exposed 

females than in the controls fed ad libitum, although 

the difference was not statistically significant: • There 

were no significant differences between the incidences 

of these neoplasms in exposed females and the weightmatched controls. - .. 

TABLE 11 

Incidences of Selected Neoplasms in Rats in the Dietary Restriction Study of t-Butyihydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

•Ad Libitum- Weight-Matched 5,000 ppm 

Fed Control Control : 

Male 

Pancreas 

Adenoma 

Overall rate a 

Adjusted~rate b 

Terminal rate c 

First incidence (days) 

Logistic regression test e 

Logistic regression test f 

J. 

Pituitary Gland (Pars Distalis) 

Adenoma 

Overall, rate 

Adjusted rate " 

Terminal rate 

First incidence (days) 

Logistic regression test 

" ~ Logigtic regression test 

Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

.Logistic regression test 

Logistic regression test 

(continued) 

3/6O (5%) 

37.5% 

3/8 (38%) 

857 (T) 

0/6O (0%) 

0.0% 

0/12 (0%) 

_d .. 

O/6O0%) 

0.0% 

0/14 (0~) 

_ 

P=0.038N 

.~ 

19/6O (32%) 

63.8% 

2/8 (25%) 

528 

19/59 (32%) 

72.4% 

7/12 (58%) 

638 

6/60 (10%) 

30.2% 

3/14 (21%) 

668 

P=0.002N 

-P=0.007N 

,. 

,,, 

19/60 (32%) 

63.8% 

2/8 (25%) 

528 

20/59 (34%) 

74.2% 

7/12 (58%) 

638 

7/60 (12%) 

31.6% 

3/14 (21%) 

627 • 

P=0.006N ' 

P~0.~08N 
Dietary IRestrictiou, NTP TR ~60 57 

TA~L~ 11 

~Incidences ol~ Selected Neop~sn~s ~u IR~ ~u.the D~et~ry IRes~r~cfiou Study o~' t-~u~ylhydroqu~none: 

Ad Libitum Feed~ug and We~gh~-Ma~ched Controls ]~hro~oco~s (continued) . 

Female 

Mammary Gland 

Fibroadenorna 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

Adenoma 

Overall rate 

Fibroadenoma or Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

Fibroadenoma, Adenoma, or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

Ad LibitumFed Control 

43/60 (72%) 

100.0% 

10/10 (100%) 

418 

3/60 (5 %) 

45/60 (75 %) 

100.0% 

10/10 (100%) 

418 

8/60 (13 % ) 

29.3 % 

1/10 (10%) 

540 

48/60 (80%) 

100.0% 

10/10 (100%) 

418 

Weight,Matched 5,000 ppm . 

Control 

23/60 (38%) 27/60 (45%) 

62.8% 74.4% 

10/22 (45%) 9/17 (53%) 

600 596 

P < 0.001N 

P=0.252 

3/60 (5 %) 2/60 (3 %) 

25/6O (42 %) 27/60 (45 %) 

66.6% 74.4% 

11/22 (50%) 9117 (53%) 

600 596 

P<0.001N 

P=0.389 

1/60 (2 % ) 4/60 (7%) 

2.1% 10.5% 

0/22 (0%) 0/17 (0%) 

729 690 

P=0.177N 

P=0.181 

26160 (43 %) 30/60 (50%) 

67.3% 76.3% 

11/22 (50%) 9/17 (53%) 

600 596 

P<.O.OOIN 

P=0.264 

(T)Terminal sacrifice 

a Number of animals with neoplasms per number of animals examined. Denominator is number of animals examined microscopically for 

pancreas, pituitary gland, and adrenal gland and number of animals necropsied for mammary gland. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

¢ Observed incidence in animals surviving until the end of the study 

d Not applicable; no neoplasms in animal group 

e In the exposed group column are the P values corresponding to the pairwise comparisons between the ad libitum-fedcontrols and the 

exposed group. The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the 

exposed group is indicated by N. 

f Result of pairwise comparison betweentheweight-matcbed controls and the exposed group 
58 Dietary Restriction; NTP TR 460 

30-MONTH RESTRICTED FEED PROTOCOL 

Survival weights of exposed rats were similar to those of the 

Estimates of 30-month survival probabilities for male controls throughout the study. 

and female rats fed restricted diets are shown in 

Table 12 and in the Kaplan-Meier survival curves in Feed Consumption, Clinical Findings, 

Figure 9. The survival rates of exposed rats were and Organ Weights 

greater than those of the controls. Feed consumption data are shown in Table K4. 

Clinical findings included hair discoloration in 

exposed males and females and urine stain on the fur 

Body Weights of exposed females.. Exposed males evaluated at 

Mean body weights of rats fed restricted diets are 3 months had greater absolute and relative liver 

shown in Figure 10 and Table J4. The mean body weights than those of the controls (Table I2b). 

TABLE 12 

Survival of Rats in the Dietary Restriction S~udy of t-Butylhydroquinone: 

30-Month Restricted Feed Protocol 

0 ppm 5,000 ppm 

• 

Male 

Animals initially in study 70 70 

3-Month interim evaluation a 10 10 

Moribund 43 33 

Natural deaths 7 5 

Animals surviving to study termination 10 22 

Percent probability of survival at the end of study b 17 37 

Mean survival (days) e 658 718 

Survival analysis d P=0.003N 

Female . : 

Animals initially in study ' 70 70 

3-Month interim evaluation a : 10 10 

Moribund ~ 39 32 

Natural deaths 3 4 

Animals surviving to study termination 18 24 

Percent probability of survival at the end of study .30 40 

Mean survival (days) 705 718 

Survival analysis . P=0.259N 

Censored from analyses 

Kaplan-Meier determinations . 

Mean of all deaths (uncensored, censored, and terminal Sacrifice) 

The result of the life table pairwise comparison (Cox, 1972) with the controls is in the exposed group column. A lower mortality in the 

exposed group is indicated by N. 
~eta~-y ~estr~ctioa, NTP TR ~60 S~ 

.... : i i : : 

1.0 ~ ~ ' ....... i ........ ! ........ !......... !.... 

~ 0.9 

0.8 

~ 0.7 09 

~ 0.6 

~ o.5 ........ ; ........ ; ........ ~"........ i......... i........ :......... ~........ ! ........ ! ........ ; ...... . ............. ~.... 

~ 0.4 

~ 0.3 

¢

~_ 0.2 i .ii ........ i... ii........

0.1 

0.0 

10 20 30 40 50 60 70 80 90 100 110 120 130 

WEEKS ON STUDY 

u .............. ; ........ ; ........ ~........ !......... ~.... 1.0 

~ 0.9 

=5 0.8 

~ 0.7 

~ 0.6 

.~ 0.5 

i FEMALERATS i i i :: ~ . 

~ 0.4 ....... ~.... RESTRICTED FEED ....... ::........ ::........ i ........ i ........ ~........ ~"" :"" 

~. o.a ....... i .... --0-- 0 ppm i i i i i i : 

0.2 

0.~ I I I I I 

10 20 30 40 50 60 70 80 90 100110 120 130 

WEEKS ON STUDY 

]~'IGU~E9 

]~aplan-IVJ~eier Survival Curves for Male and ~e~na~e ~ats Achnin~stered ~t-Buty~hydroquinone 

~n Feed for 30 IV~onths: Restricted ~'eed Protocol 
-. . 

~) Dietary Restriction, NTP TR 460 

45O ! • : i i ! 

~ ' " ' ~ -e.o.~ ............. i............ ~ 400 ............ i ............. i ........ ~0~~o~oO0~~eo :1~ [] [] i . : 

i. joo i i i° i ~OOo~ ~ 350 ............ ~OoO ......................... ~............. ~............. : ............ :......... oo-O...... 

z ............. ~........................................ , ...... : ...... ~................................................ ~... 3o0 

~5o .... ........................................................ - ................. ~-~ ..... ~............- -

~ ~oo ...... ~ ~:.........................~............~:.....~...................~..........:.~........................ 

......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....... 

FEED 

~ ...... , ..... ............ ............ ............. ...... ~ ~oo ........... ~ ~ ~,~o~ ' 

50 ~ ~ ~ ~ ,. 

0 15 30 45 60 75 90 105. 120 

WEEKS ON STUDY 

., 

,. 

400 ~ : 

; 

~ { = 3~0 q ............ ~............. ~............ ~............ ~............. ~............. ~......................... ~............ 

< ~ ~ ~ ~ ~ o~OOo~ ~ 4 ............. ~ ~~~ ~ ~ 300 ....... " ............................................................... ~ ~ ~ ~ ~ ~8-~-~ 

~ ~ ~ :: ~ :: ~ @o: ' ~ ~ 0 

~ = 250 / :: ~ oo~o 0~°0~° ~ ~ ..... 

~ .200 J ............. ~............. ~~ ......... ~...o....4 ............. ~............. ; ............ 4............. ~................ 

o ~o °° ~ ~ o ~ ~ 1 

~ ~s0 -;~.............~ ~............. ~............~.............~--~. . . . . . . . ~ ~ z /~ ~ ~ ~ ~ " ~'" ~" ~ESTRIC~E o FEE~ I 1 

< ~. .......... ~............. :............. :............ ;....... ~..... :...... ~ op~e . I " ~ 100 

= ~0~ ' . . , ~"~°~T., , ~I, 

0 15 30 45 60 75 90 105 120 

WEEKS ON STUDY ' 

. , 

FIGURE 10 

Growth Curves for Male and Female Rats Administered t-Butylhydroquinone 

in Feed for 30 Months: Restricted Feed Protocol 
I 

Dietary Restriction, NTP TR 460 61 .. 

Pathology and Statistical Analyses 

This section describes the statistically significant or are presented in Appendix C for male rats and Appenbiologically noteworthy changes inthe incidences of dix D for female rats. 

neoplasms of the clitoral gland, mammary gland, and 

pituitary gland. Summaries of the incidences of Clitoral Gland: Exposed females had significantly 

neoplasms and nonneoplastic lesions and statistical greater incidences of clitoral gland adenoma and 

analyses of primary neoplasms that occurred with an adenoma or carcinoma (combined) than those in the 

incidence of at least 5 % in at least one animal group control females (Tables 13 and D2b). 

'FABLE 13 

Incidences of Clitoral Gland Neoplasms in Female Rats in the Dietary Restriction Study 

of t-Butylhydroquinone: 30-Month Restricted Feed Protocol 

0 ppm 5,000 ppm 

Adenoma 

Overall rate a 3/59 (5 % ) 10/59 (17 %) 

Adjusted rate b 13.4% 27.5% 

Terminal rate c 1/18 (6%) 3/23 (13%) 

First incidence (days) 866 673 

Logistic regression test d P=0.044 

Carcinoma 

Overall rate 2/59 (3%) 5/59 (8%) 

Adjusted rate 6.2% 21.7% 

Terminal rate 0/18 (0%) 5/23 (22%) 

First incidence (days) 788 911 (T) 

Logistic regression test P=0.279 

Adenoma or Carcinoma 

Overall rate 5/59 (8 %) i5/59 (25 %) 

Terminal rate 18.8 % 45.6 % 

Adjusted rate 1/18 (6%) 8/23 (35%) 

First incidence (days) 788 673 

Logistic regression test P=0.019 

(T)Terminal sacrifice 

a Number of animals with-neoplasms per number of animals with clitoral gland examined microscopically 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

e Observed incidence in animals surviving until the end of the study 

d In the exposed group column are the P values 9orresponding to the pairwise comparisons between the controls and the exposed group. The 

logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. .. 
62 Dietary Restriction, NTP TR460 

Mwnmary Gland: Exposed females had significantly Pituitary Gland: :The incidence, of adenoma of the 

lower incidences of fibroadenoma and fibroadenoma pars distalis in exposed females was significantly less 

or carcinoma (combined) than did the controls than in the controls (Tables 14 and D2b). 

(Tables 14 and D2b). . i 

TABLE 14 

Incidences of Selected Neoplasms in Female Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

30-Month Restricted Feed Protocol 

Mammary Gland 

Fibroadenoma 

Overall rate a 

Adjusted rate b 

Terminal rate c 

First incidence (days) 

Logistic regression test d 

Fibroadenoma or Carcinoma 

Overall .rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Pituitary Gland (Pars Distalis) 

Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

.. 

0 ppm 5,000 ppm 

30/60 (50 % ) 17/60 (28 %) 

86.8 % 56.1% 

14118 (78%) 12/24 (50%) 

688 '~ 780 

P=0.002N 

31160 (52%) 17160 (28%) 

87.5% 56.1% 

14/18 (78%) 12/24 (50%) 

688 780 

P<0,001N 

30/59 (51%) 18/60(30%) 

81.1% 50.8% 

11/17 (65%) 9/24 (38%) 

638 649 

P=0,007N 

a Number of animals with neoplasms per number of animals examined. Denominator is number of animals examined microscopically for 

pituitary gland and number of animals necropsied for mammary gland. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intereurrent mortality 

c Observed incidence in animals surviving until the end of the study 

d In the exposed group column are the P values corresponding to the pairwise comparisons between the controls and the exposed group. The 

logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the exposed gioup is 

indicated by N. 
Dietary ll~estriction, ~TP TR ,~6~ ~.3 

AL,I C OL,AZ ULFAI r I HE ( 'AVAGE 

ATS 

t~D L~BITUM ]~'EED~NG AND WE~GHT-I~TCHED CONTROLS ~>I~OTOCOLS 

Sur~i~,al 

Estimates of 2-year survival probabilities for male rats 

fed ad libitum and the weight-matched control rats are 

shown in Table 15 and the Kaplan-Meier survival 

curves in Figure 11. The survival of dosed males was 

less than that of the controls fed ad libitum and 

slightly less than that of the weight-matched controls. 

 ody We ghes 

Mean body weights are given in Figure 12 and in 

Table J5. Because there was negligible body weight 

depression in the 337.5 mg/kg group throughout the 

study, no adjustments were made to the amount of 

feed presented to the weight-matched controls, 

thereby yielding a redundant control group. At 

1 year, the mean body weight of dosed males was 

TABLE 15 

96% that of the controls fed ad libitum and 98% that 

of the weight-matched controls. During the second 

year of the study, the mean body weight of dosed 

males remained within 5 % of the mean body weights 

of both control groups. 

Feed Consumpt~an, Clinical ~ndings, 

and Organ Weights " 

Feed consumption by dosed males was similar to that 

by the control groups (Table K5). There were no 

clinical findings considered related to treatment with 

salicylazosulfapyridine. The absolute spleen weight 

of dosed males was significantly less than that of the 

controls fed ad libitum; there were no significant 

differences in organ weights between dosed males and 

the weight-matched controls (Table I3a). 

Survlva] of Male Rats in the Dietary Restriction Study of Sa~icy~azosulfapyr~ne: 

Ad Libitum Feeding and Weight-Matched Controls ~*rotoco~s 

Ad Libit~m- Weight-Matched 337,5 mg/kg 

Fed Control ~• Control 

Animals initially in study 70 

6-Month interim evaluation a 10 0 0 

15-Month interim evaluation a 10 10 10 

Accidental deaths a I 2 2 

Moribund 13 16 15 

Natural deaths 1 1 10 

Animals surviving to study termination 35 31 23 

Percent probability of survival at the end of study b 71 65 48 

Mean survival (days) e 582 641 629 

Survival analysis d P=0.036 

Survival analysis e P=0.170 

Censored from analyses 

Kaplan-Meier determinations 

Mean of all deaths (uncensored, censored, and terminal sacrifice 

The result of the life table pairwise comparison (Cox, 1972) with the controls fed ad libitum is in the dosed group column. 

Result of life table palrwise comparison with the weight-matched controls 
Dietary Restriction, NTP TR 460 

1.00 

0.95 

0.90 

~. 0~85 

,~@ 0.80 

~ 0.75 

~ 0.70 _l 

~ 0:65 .......... . ........... ; ........... . ........... ~....~ ..... ,... 

< 

8 0.60 ~ ~ Ad Libitum-Fed Control ............................................. : .......... i i 

~" :0.55 

0.50 ~ 337.5,mg~g " 

0.45 --'" ........................... ' .............. : ................... i ........ :":" .......... : ..................... i .... 

0:40 • " : ; : 2 2 : 

10 20 aO 40 50 60 70 80 90 100 

WEEKS ON STUDY 

FIGURE 11 

Kaplan-Meier Survival Curves for Male Rats Administered SalicYlazosulfapyridine in Corn.oil 

by Gavage for 2 Years: Ad Libitum Feeding and Weight-Matched Controls Prot0cols,. • 

500 

4~o ................ ~...... ~....... ~--..-,:~-~--=-~ ....... ~ ............. i " '~~~=~~ 

< 

~: 400 ........ ,.~i:i ..... ,~:.~.~.~° ........... i ............... ~i ........ " ....... i i ............... ........... ~@t L? 

Z 350 ................ i -''B~ ...... !": ....... ,"'i I ............. ! ............... !...:....1,....:!.....,:., ..... ........ I 

...... ...... ..... ..... ..................... . ...... ....... ............ ~(..9 300 -

..... ~ ....... ~ ..... ; ......... ~: ..... ~....' ...... i ............... ~ ............... ~-: .......................... ---= .... ~ > 250-

..~ ..... ~,...~ ............................... ~--r ............ ~ ............................ ~ ........................ 

~ 200- z 

< 

w 150. ................ ~.............. : .................. ~............... ~"" ~ 'Ad Libitu~Fed Control,

~ • " . ~ Weight-Matched Control 

100-~ ............. ~ ......... : ........................................... • ~ zzzs=~ 

~ ~ 

~ r I' ~ 

0 " 15 30 45 60 75 90 105 

WEEKS ON STUD7 ., 

FIGURE 12 ~ 

Growth Curves for Male Rats Administered Salicylazosulfapyridine in Corn Oil by Gavage 

for 2 Years: Ad Libitum Feeding and Weight-Matched Controls Protocols 
Dietary Restriction, NTP TIt 460 65 

zUathology and Statistical Analyses rats in the salicylazosulfapyridine study are provided 

This section describes the statistically significant or in NTP Technical Report 457 (NTP, 1997b). 

biologically noteworthy changes in the incidences of 

mononuclear cell leukemia and of neoplasms and/or Urinary Bladder: The incidence of urinary bladder 

nonneoplastic lesions of the urinary bladder, kidney, papilloma was significantly greater in dosed males 

and spleen. Summaries of the incidences of neo- than in the controls fed ad libitum or the weightplasms and nonneoplastic lesions and statistical matched controls (Tables 16 and E2a). Two dosed 

analyses of primary neoplasms that occurred with an males had multiple papillomas; no papillomas 

incidence of at least 5 % in at least one animal group occurred in either control group. Additionally, the [ : are presented in Appendix E. Pathologic descriptions incidences of hyperplasia of the urinary bladder 

of neoplasms and nonneoplastic lesions occurring in mucosa, concretion, dilatation, and grossly diagnosed 

TABLE 16 

Incidences of Neoplasms and Nonneoplastic Lesions of the Urinary Bladder and Ifddney 

in Ma~e ]Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum ~'eeding and Weight-Matched Controls Protocols 

Ad Libitum- Weight-Matched 337.5 mg/kg 

Fed Control Control 

Urinary Bladder a 50 50 50 

Concretion b 0 0 10**AA 

Dilatation 0 1 7*A 

Mucosa, Hyperplasia 0 0 41**A~ 

Calculus (gross diagnosis) c 0 0 27"*A~ 

Papilloma d 

Overall rate e 0/50 (0%) 0/50 (0%) 6/50 (12%) 

Adjusted rate f 0.0% 0.0% 22.1% 

Terminal rate g 0/35 (0%) 0/31 (0%) 3/23 (13%) 

First incidence (days) . _h _ 653 

Logistic regression test 1. P=0.011 

Logistic regression testd P=0.013 

Kidney 50 50 50 

Concretion 0 0 33"*A~ 

Hydronephrosis 0 0 28"*~ ~ 

Mineralization 3 6 13"* 

Renal Tubule, Dilatation 0 1 11"*~ 

Transitional Epithelium, Hyperplasia 10 5 43"*A~ 

* Significantly different (P~0.05) from the ad libitum-fed controls by the logistic regression test 

** Pg0.01 

A Significantly different (P~0.05) from the weight-matched controls by the logistic regression test 

AA P~0.01 

a Number of animals With tissue examined microscopically 

b Number of animals with lesion 

¢ Number of animals with grossly observed urinary bladder concretions (diagnosed as calculi at necropsy) 

d Historical incidence for ~2-year NTP gavage studies with control groups receiving corn oil by gavage and fed ad libitum (mean ::1: standard 

deviation): 1/904 (0.1% ± 0.5%); range, 0%-2%. 

e Number Of animals with neoplasms per number of animals with tissue examined microscopically 

f Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

g Observed incidence in animals surviving until the end of the study 

h Not applicable; no neoplasms in animal group 

In the dosed group column is the P value corresponding to the pairwise comparison between the ad libitum-fed controls and the dosed 

group. The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. 

J Result of pairwise comparison between the weight-matched controls and the dosed group 

i 
66 Dietary Restriction, NTP TR 460 

calculi in dosed males were greater than those in pigmentation (14/50, 20/50, 30/50) than the controls 

either control group (Tables 16 and E3a). fed ad libitum or the weight-matched controls 

(Table E3a). The greater incidences of these lesions 

Kidney: Dosed males had significantly greater in dosed males are suggestive of erythrocyte toxicity 

incidences of concretions, transitional epithelial and destruction (hemolysis).. 

hyperplasia, hydronephrosis, mineralization, and renal 

tubule dilatation than the controls fed ad libitum or the Mononuclear Cell Leukemia: Dosed males had a 

weight-matched controls (Tables 16 and E3a). significantly lower incidence of mononuclear cell 

leukemia than the controls fed ad libitum (Tables 17 

Spleen: Dosed male rats had significantly greater and E2a). The incidence in dosed males was also 

incidences of hematopoietic cell proliferation lower than that in the weight-matched controls; 

(ad libitum-fed controls, 14/50; weight-matched however, the difference was not statistically 

controls, 9/50; 337.5 mg/kg,.23/50) and hemosiderin significant. 

,.-

TABLE 17 • 

Incidences of Mononuclear Cell Leukemia in Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols 

- Ad Libitum- Weight-Matched 337.5 mg/kg 

Fed Control Control 

Mononuclear Cell Leukemia a 

Overall rata b 13/50 (26%) 10/50 (20%) 3/50 (6%) 

Adjusted ratec 32.0% 26.2% 6.8% 

Terminal rate d 8/35 (23%) 5/31 (16%) 0/23 (0%) 

First incidence (days) 477 526 428 

Life table test e P=0.040N 

Life table test f P=0.076N 

a Historical incidence for 2-year NTP gavage studies with control groups receiving corn oil by gavage and fed ad libitum (mean ± standard 

deviation): 224/922 (24.3% ± 10.2%); range; 10%-46%. Includes lymphocytic, monocytic, mononuclear ceil, and undifferentiated 

leukemias. 

b Number of animals with neoplasms per number of animals necropsied 

c Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

d Observed incidence in animals surviving until the end of the study 

e In the dosed group column is the P value corresponding to the pairwise comparison between the at/libitum-fed controls and the dosed 

group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. A 

lower incidence in the dosed group is indicated by N. 

f Result of pairwise comparison between the weight-matched controls and the dosed group 
67 

c 

Dietary ~estr~cfion, NTP TS ~ 

2-YEAR AND 30-MONTH ]~EST~,~CTED ]~'ESD ]~]I~DTOCOLS 

Survi~,al 

Estimates of 2-year and 30-month survival probabilities for male rats fed restricted diets are shown in 

Table 18 and the Kaplan-Meier survival curves in 

Figure 13. The survival of dosed rats was greater for 

the controls at 2 years and at 30 months. Survival in 

the control group was reduced to 20 % at week 130. 

1Early Weights 

Mean body weights are shown in Figure 14 and in 

Table J6. The mean body weights of dosed males 

were 94 % those of the respective controls at 1 year. 

The mean body weights of dosed males ranged from 

6 % to.. 18 % less than the mean body weights of the 

controls during the second year of the study. From 

2 years to 30 months, the mean body weight of dosed 

males ranged from 16% to 20% less than that of the 

controls. ~ 

Feed Cansumpt~on, Clinical ~ndings, 

and Organ Weights 

Feed consumption data are shown in Table K6. There 

were no clinical findings considered related to treatment with salicylazosulfapyridine. At the 15-month 

evaluation, there were no significant differences in 

organ weights between dosed and control males 

(Table I3b). 

TABLE 18 

Survival of Male Rats in the Dietary Reestriction Study of Salicynazosu~fapyridhae: 

2-Year and 30-Month Restricted ~'eed ]Protocols 

2-Year Restricted Feed 30-Month Restricted Seed. 

Vehicle 337.5 mg/kg Vehicle 337.5 mg/kg 

Control Control 

Animals initially in study 61 60 49 50 

15-Month interim evaluation a I0 10 - 0 0 

Accidental deaths a 2 2 4 2 

Moribund 13 6 28 14 

Natural deaths 2 3 7 I0 

Animals surviving to study termination 34 39 10 24 b 

Percent probability of survival at the end of study c 70 82 22 50 

Mean survival (days) d 658 642 765 798 

Survival analysis e P=0.313N P=0.012N 

a Censored from analyses 

b Includes one animal that died during the last week of the study. 

Kaplan-Meier determinations 

d Mean of all deaths (uncensored, censored, and terminal sacrifice) 

e The result of the life table pairwise comparison (Cox, 1972) with the controls is in the dosed group column. A lower mortality in a dosed 

group is indicated by N. 
Dietary Restriction, NTP TR 460 

~.o( =: ! • ~ 1 ..~ i~ -" .............. .: ............ ! ............ i!i ........... 

i 0.6 " ' ,0.5 ............. :~." "': "MALE RATS ' " .. 

" i • ~.v~,~cl~ co,t,~(.2 ,r) i • i . i Y ~ /

0.4 ............. i...: ~ V~hicl~Co.Uol(30mo) ~ ...... : ...... fl 

i • 337.5 mg/kg (2 yr) i ! ~ I / .... :....:...:, .... ~ 337.5 mgm9 (30 too) ......... i............. !............. i ............. i.. .......... 

o~ ~ ........ .i,:-: ......... ~...... :,~ ...... ': ....... ~1 

0.2 i i i i 

I I I I I I 

" 15 30 45 60 75 :. 90 1.05 120 

WEEKS ON sTuDY " 

FIGURE 13 

Kaplan:Meier Sur~vival Curves forMal e Rats Administered Salicylazosulfapyridine in Corn Oil 

- ~ ~by Gavage for up to 30 Monti~: 2-Year and ~0-M0nth Restricted Feed protocols 

• 

• 

• 

• 

• 

, ,.. ,~..~ . .,~' ., 

, 

5OO / i 

-4 ..... ~.......................................... i_.i ........... . ........ .. ................ ..................... 450 / . i '! ! ~o~o! i i i ~ ............ i.: ....... .:..:: ............. :~.~ ...... 0..: .......... :..O.gO.dO.O._e ~ ............. i............ i 400 -I i i n~ o~z~i : U~n~ i 

350 / i ~:no~ i ~= i :;- oh 

-I ............ : ...... ~O!,5~v : ............ : ............. ig~~i~~ ...... ! ............ b,= ....... 

| ::nO ~ Oi i :: :: :: ~ ::u 

300 .......... ~8-~ ........ i............. ! ............ i............. i............. ; ............ i.~..~,.~..i ...... [] .... 

o 200 i:i .......... ............ , ............ . . . . 

Z • Vehicle Control (2 yr) 

~ 150 . " [] Ve"icle Co.tr0~30~o) • 337.5 mg/kg (2 yr) 

~ 1 00 ~ 337.5 mg/kg (30 TO) ° f 

50 ~ ~ i 

I I I . 

0 15 30 45. 60 75 90 105 120 

,WEEKS ON STUDY 

• ' ~ 

F]6tmE ]4 

Growth Curves f~r Male Rats Administered Salicylazosulfapyridine in Corn Oil by Gavage 

for up to 30 Months: 2-Year and 30-Month Restricted ~Feed Protocols 
Dietary Restriction, NTP TR 4~ 69 

z~a~hology and Statistical Analyses 

This section describes the statistically significant or 

biologically noteworthy changes in the incidences of 

mononuclear cell leukemia and of neoplasms and/or 

nonneoplastic lesions of the urinary bladder, kidney, 

spleen, and adrenal medulla. Summaries of the incidences of neoplasms and nonneoplastic lesions and 

statistical analyses of primary neoplasms that occurred 

with an incidence of at least 5 % in at least, one animal 

grou~ are presented in Appendix E. 

Urinary Bladder: The urinary bladder effects 

observed in dosed male rats fed ad libitum did not 

occur in males fed a restricted diet for 2 years or 

30 months. Hyperplasia of the mucosal transitional 

epithelium was the only urinary bladder lesion that 

occurred with a significantly greater incidence in 

dosed males fed restricted diets than in the controls at 

2 years or at 30 months (Tables 19 and E3b). No 

papillomas of the urinary bladder occurred in dosed 

males at 2 years; at 30 months, one dosed male had a 

papilloma, and concretions (diagnosed as calculi) were 

observed grossly in four dosed males at necropsy. 

Kidney: The incidences of concretion and transitional 

epithelial hyperplasia were significantly greater in 

dosed males at 2 years and at 30 months than in the 

controls (Tables 19 and E3b). Dosed males also had 

a significantly greater incidence of mineralization than 

the controls at 2 years. The incidence of nephropathy 

in dosed males was significantly greater than that in 

the controls at 30 months. 

Spleen: Dosed males had significantly greater incidences of hen~osiderin pigmentation than the controls 

at 2 years (vehicle control, 12/51;.337.5 mg/kg, 

35/50) and at 30 months (15/49, 33/49) (Table E3b). 

TABLE 19 

Incidences of Neopi~sms and Nonneopl~stic Lesions in the Urinary Bnadder and ~dney . 

in Male Rats in the Dietary Restriction Study of SaaicyRazosulfapyridine: 

2-'Year and 30-Month Restricted Feed Protocols 

2-Year Restricted Feed 30rMonth Restricted Feed 

Vehicle 337.5 mg/kg Vehicle 337.5 mg/kg 

Control Control 

Urinary Bladder a 51 50 ~9 49 

Mucosa, Hyperplasia b 0 7** 0 8** 

Calculus (gross diagnosis) c 0 0 0 4 

Papilloma 0 0 0 1 

Kidney 51 50 49 50 

Concretion 0 . 22** 0 35** 

Mineralization 2 11"* 4 7 

Nephropathy 44 46 39 48* 

Transitional Epithelium, Hypetrplasia 3 18"* 1 37** 

* Significantly different (P~0.05) from the control group by the Fisher exact test 

** P~0.01 

a Number of animals with tissue examined microscopically 

b Number of animals with lesion 

¢ Number of animals with grossly observed urinary bladder concretions (diagnosed as calculi at necropsy) 
70 Dietary Restriction, NTP TR 460 

Mononuclear Cell Leukemia: At 2 years and at Adrenal Medulla: The incidences of benign 

30 months, the incidences of mononuclear cell leuke- ~pheochromocytoma and benign or malignant pheomia in dosed males were significantly less than those chromocytoma (combined) were significantly less in 

in the controls (Tables .20 and E2b). dosed rats than in the controls at 2 years and at 

30 months (Tables 20 and E2b). 

TABLE 20 

Incidences of Selected Neoplasms in Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle 337.5 mg/kg Vehicle 337.5 mg/kg 

Control Control 

Mononuclear Cell Leukemia 

Overall rate a 11/51 (22 % ) 2/50 (4 %) 24/49 (49 %) 8/50 (16 %) 

Adjusted rate b 43.4% 6.9% 71.3% 23.8% • 

Terminal rate c 3/34 (9%) 2/39 (5%) 3/10 (30%) 3/24 (13%) • 

First incidence (days) 520 731 (T) 556 430 

Life table test d P=0.009N P<0.001N 

Adrenal Medulla ~ :~" 

Benign Pheoehromocytoma 

Overall rate 8/51 (16%) 1/50 (2%) • 14/48 (29%) 8/50"(16%) 

Adjusted rate 22.2% 3.4% 66.7% ' 31.1% 

Terminal rate 6/34 (18%) 1/39 (3%) 4/I0 (40%) 7/24 (29%) 

First incidence (days) - 700 731 (T) 752 812 

Logistic regression test d P=0.014N P=0.011N 

Malignant Pheochromocytoma 

Overall rate 2/51 (4%) 0/50 (0%) 1/48 (2%) 1/50 (2%) 

Benign or Malignant Pheochromoeytoma 

Overall rate 9/51 (18%) 1/50 (2%) 15148 (31%) 8/50 (16%) 

Adjusted rate 33.3 % 3.4 % 72.3 % 31.1% 

Terminal rate 7/34 (21%) 1/39 (3%) 5/10 (50%) 7/24 (29%) 

First incidence (days) 700 731 (T) 752 812 

Logistic regression test P=0.007N P=0.005N 

(T)Terminal sacrifice 

a Number of animals with neoplasms per number of animals examined. Denominator is number of animals examined microscopically for 

adrenal medulla and lung and number of animals necropsied for mononuclear cell leukemia. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intereurrent mortality 

e Observed incidence in animals surviving until the end of the study 

d In the dosed group eolunms are the P values corresponding to the pairwise ~omparisons between the controls and that dosed group. The 

life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. For all tests, a lower incidence in a dosed group is indicated by N. 
Dietary Restriction, NTP TR 4~0 71 

taD LIBITUM ~SD~NG AND WE~GHT-]V~ATC~ED CONTROLS ~oTDcoLS 

Survival mean body weight of dosed males ranged from 10% 

Estimates of 2-year survival probabilities for male to 17% less than that of the controls fed ad libitumbut 

mice fed ad libitum and the weight-matched controls remained within 10% that of the weight-matched 

are shown in Table 21 and the Kaplan-Meier survival • controls. 

curves in Figure 15. The survival rate of mice in the 

2,700 mg/kg group was slightly greate~ than that of Feed Consumption, Clinical Findings, the controls fed ad libitum and was similar to that of 

and Organ Weights the weight-matched controls. 

Dosed males consumed more feed than the controls 

Body Weights fed ad libitum (Table K7). There were no clinical 

Mean body weights are shown in Figure 16 and findings considered related to salicylazosulfapyridine 

Table J7. The mean body weight of dosed males was administration. The absolute and relative liver and 

17% less than that of the controls fed ad libitum and spleen weights of dosed males were significantly 

1% less than that of the weight-matched controls at greater than those of the weight-matched controls 

1 year. During the second year of the study, the (Table I4a). 

TABLE 21 

Survival of Male Mice in the Dietary Restriction Study of Salicygzzosulfapyridine: 

Ad Libitum Feeding and Welght-lV~atched Controls Protocols 

Ad Libitum- Weight-Matched 2,700 mg/kg 

Fed Control Control 

Animals initially in study 60 60 60 

15-Month interim evaluation a 10 10 10 

Accidental deaths a 2 l 0 

Moribund 5 3 4 

Natural deaths 3 1 0 

Animals surviving to study termination 40 45 46 

Percent probability of survival at the end of study b 84 92 93 

Mean survival (days) c 649 665 653 

Survival analysis d P=0.350N 

Survival analysis e P= 1.000 

a Censored from analyses 

b Kaplan-Meier determinations 

Mean of all deaths (uncensored, censored, and terminal sacrifice) 

d The result of the life table pairwise comparison (Cox, 1972) with the controls fed ad libitum is in the dosed group column. A lower 

mortality in the dosed group is indicated by N. 

e Result of life table palrwise comparison with the weight-matched controls 

c 
72 Dietary Restriction, NTP TR 460 

,.oo I ' 1 ~ ,.Li .........i........ 

i i. i! 

O 

o.~ ......................... ~..... i .......... i ...... ~................................. ~........ '....... ~ ..... . ....... ~t- : : : , , . 

m 

MALE MICE 

O ~ Ad Libitum-Fed Control 0"80 t 

~ Weight-Matched Control 

~ 2,700 mg/kg ...... ..... ........... , ........... ~ .......... ; ................... 0.7 5 : 

0.70 : 

10 20 30 40 50 60 70 80 . 

90 100 

WEEKS ON STUDY 

FIGURE 15 

Kaplan-Meier Survival Curves forMale Mice Administered Salicylazosulfapyridine in Corn Oil 

by Gavage for 2 Years: AdLibitum Feeding and Weight-Matched Controls Protocols 

00 _~ ~. ,,~ ~ 

~ 55~. ......... : ..... ":: .............................................. i i ooi. O°~'~o*Ooo ................................................. i

5o -I ............... i............... i ....... ~;..~..~,..o ......... i ............. ~ ............... ~~~~::; / : : • : :~ ~ ~ ~ 

i : i ~ ~'~"~-~.~ : ~ . ~ : - 45 ~ ............... , .......... ~-~ ............... ~ .......... B~ ......... ~-~.~-.~ .... ~....... ~ ......~..... ~ : : ~ : : ~ : :

.I :: ~ ~ :: :: :: . ~ ............... ...... ...... ............... ............... ............... ......... ........... 

~ :~ ~ ~ ~ ~ 

~ . : : • 

35 ............ ..---~:-~ ............ ................ ~..-~ .......... ~ ~ ............... ': ............... ': ........... .-~ ..... 

30 .,.~ ............... ~ ................ ~............... ~.... ~ ~c~ ......... 

~ ~ :: ~ ~ ~ AdLibitum-FedControl : 

~ 25 4~ ........... ; ................ :~ ................ ~:................... ~ w~,~-~=~co,~,o~ ...~. .... 

~ ~ ~ 2700mg~g I :: 

20 ~ ~ ~ ~ ~ ' 

0 15 " 30 45 60 75 90 105 

WEEKS ON STUDY 

FIGURE 16 

Growth Curves for Male Mice Administered Salicylazosuifapyridine in Corn Oil by Gavage 

for 2 Years: Ad Libitum Feeding and Weight-Matched Controls Protocols 
D~e~ary Restriction, NTP TR 460 73 

Pathology and Statistical Analyses 

This section describes the statistically significant or 

biologically noteworthy, changes in the incidences of 

neoplasms and nonneoplastic lesions of the liver and 

spleen. Summaries of the incidences of neoplasms 

and nonneoplastic lesions and statistical analyses of 

primary neoplasms that occurred with an incidence of 

at least 5 % in at least one animal group are presented 

in Appendix F. Pathologic descriptions of neoplasms 

and nonneoplastic lesions occurring in mice in the 

salicylazosulfapyridine study are provided in NTP 

Technical Report 457 (NTP, 1997b). 

Liver: Dosed mice had significantly greater incidences of hepatocellular adenomas and hepatocellular 

adenomas or carcinomas (combined) than the controls 

fed ad libitum or the weight-matched controls 

(Tables 22 and F2a). Additionally, the incidences of 

clear cell and eosinophilic foci were significantly 

greater in dosed males than in the ad libitum-fed and 

weight-matched control groups. However, the incidence of hepatocellular carcinomas in dosed males 

was slightly less than in the controls fed ad libitum. 

Spleen: Dosed male mice had a significantly greater 

incidence of hemosiderin pigmentation than the 

ad libitum-fed or weight-matched controls (ad libitumfed controls, 2/50; weight-matched controls, 1/50; 

2,700 mg/kg, 47/50; Table F3a). The incidence of 

hematopoietic cell proliferation in dosed males was 

also significantly greater than that of the weightmatched controls (3/50, 13/50). 
74 

c 

Dietary Restriction, NTP TR 460 

']'ABLE 22 

Incidences of Neoplasms and Nonneoplastic Lesions of the Liver in Male Mice 

in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

Number Examined Microscopically " 

clear Cell Focus a 

Eosinophilic Focus 

'Hepatocellular Adehoma 

Overall rate b 

Adjusted rate e 

Terminal rate d 

First incidence (days) 

Logistic regression test e 

Logistic regression test f . 

Hepatocellular Adenoma, Multiple 

Overall rate 

Hepatocellular Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regressio 9 test 

Logistic regression test 

• 

Hepatocellular Adenot~a or Carcinoma g • 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

Ad ILibitumFed Control 

50 

2 

6 

13/50 (26%) 

32.5% 

13/40 (33%) 

728 (T) 

1/50 (2%) 

13/50 (26%) 

29.2% 

9/40 (23 % ) 

420 

24/50 (48%) 

54.3% 

20/40 (50%) 

420 

Weight-Matched 2,700 mg/kg 

Control 

50 50 

2 11,,, 

1 22"*~, 

8•50 (16%) 42/50 (84%) 

17.8% 87.5% 

8/45 (18%) . 40/46 (87%) 

728 (T) 497 

P<0.001 

P<0.001 

0/50 (0%i • 27/50 (54%) 

6/50 (12%) 8/50 (16%) 

12.9% 17.4% 

5/45 (11%) 8/46 (17%) , 

574 728 (T) 

P=0.159N 

P=0.378 

14/50 (28%) 44/50 (88 %) 

30.3% 9i.7% 

13/45 (29%) 42/46 (91%) 

574 497 

P<0.001 

P<0.001 

(T)Terminal sacrifice 

* Significantly different (P~0.05) from the ad libitum-fed controls by the logistic regression test 

** P~0.01 

s, Significantly different (Pa0.01) from the weight-matched controls by the logistic regression test 

a Number of animals with lesion 

b Number of animals with neoplasms per number of animals with liver examined microscopically 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

d Observed incidence in animals surviving until the end of the study 

e In the dosed group column are the P values corresponding to the pairwise comparisons between the ad libitum-fed controls and the dosed 

group. The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the dosed 

group is indicated by. N. 

f Result of pairwise comparison between the weight-matched controls and the dosed group 

g Historical incidence for 2-year NTP garage studies with control groups receiving corn oil by gavage and fed ad libitum (mean 5: standard 

deviation): 340/763 (44.6% 5: 14.6%); range, 25%-70%. 
c 

Dietary ]~estriction~ ~ TR ~60 7S 

2-~EA~ AND 3-'~'EAR ]~EST~CTED ~'EED ~OTOCOLS 

Survival 

Estimates of 2-year and 3-year survival probabilities 

for male mice fed restricted diets are shown in 

Table 23 and the Kaplan-Meier survival curves in 

Figure 17. The survival of dosed mice was similar to 

that of the controls at 2 years but significantly greater 

than that of the controls at the end of the study. 

 ody Weighes 

Mean body weights are shown in Figure 18 and 

Table J8. The mean body weights of the two groups 

of dosed mice were about 80% those of the respective 

controls at 1 year. During the second year of the 

study, the mean body weights of the dosed males 

ranged from about 71% to 82% those of the controls; 

after 2 years, the mean body weight of dosed males 

recovered slightly, increasing to 89% that of the 

controls by the end of the study. 

Feed Consump~on, Clinical ~ndings, 

and Organ Weggh~s 

Feed consumption data are shown in Table KS. 

Dosed mice fed restricted diets had diarrhea; no other 

clinical findings were considered related to chemical 

administration. For dosed males, the absolute right 

kidney weight was significantly less than that of the 

controls, and the relative right kidney weight and 

absolute and relative spleen weights were significantly 

greater than those of the controls (Table I4b). 

TABLE 23 

Survival of Male lV~ce ~n the Dietary ~estriction Study of Sa~icy]azosu~fapyr~e: 

2-'Year and 3-Year ~estr~cted Feed ~otocols 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle 2,700 mg/kg Vehicle 2,700 mg/kg 

Control Control 

Animals initially in study 62 60 48 50 

15-Month interim evaluation a 10 10 0 0 

Moribund 6 5 21 9 

Natural deaths 4 1 7 7 

Animals surviving to study termination 42 44 20 34 

Pereem probability of survival at the end of study b 81 88 42 68 

Mean survival (days) c 672 665 936 1,003 

Survival analysis d P=0.507N P=0.010N 

a Censored from analyses 

b Kaplan-Meier determinations 

Mean of all deaths (uncensored, censored, and terminal sacrifice) 

d The result of the life table pairwise comparison (Cox, 1972) with the controls is in the dosed group column. A lower mortality in a dosed 

group is indicated by N. 
76 Dietary Restriction, NTP TR 460 

• ~:, 

..... 

1.00 ~': :" ~ ~ ~~ililll 0.95 

~ 0.90 

0.85 

~ 0.80 ~f~ 

b 0.75 

~ 0.70 

- •0.65 

~< 0.60 

1 ........ ...... :..... ........i..... o~ 0.55 

a. 0.50 :~::::::::I~v??i'i0,~?'i?'i',)"l::::::::::::: .......~ ~.~ ~ ~.~.:::~::::: 

0.45 ~ ....... ~ ~ ~ ................... ..... ........... ...... ....... -~ ....................~...........~,~......~..~.............~.% 

0.40 I I I .~.. I I I . . 

~5 aO 45 60 75 .90. ]05 ~20 a5 ~50 . 

. 

"' WEEKS ON STUDY . 

, 

, 

FIGURE 17 

Kap|an-Meier Survival Curves for Male Mice Administered Salicylazosulfapyridine in Corn Oil 

by Gavage for 2 or 3 Years: 2-Year and 3~-YearRestricted Feed Protocols : 

. . ~ 

• 

' ~ i 

55 

50 .......... :~ .......... ~: .......... : ........ ~:~.=-~=.~ ......... :::..-.'..i: ':o .......... ~:..-~ ...... ~ ........... ~.......

~ 4o 

:~ 45 • ......... :,.......... ;.=0..; ........... ~:..... ~1~1~ ...... Ll~o~.~.gooi-~ ........... ~..... ::....... 

:: ::~ ~i ~ ~ ~ ~=~ Ooos " ...........::.........0.........O0......::..........~ ......o.~.... ......~..........~..........:oo~:. ....... 

~ as 

~ ao 

25 •. , 

~ 20 

o 

m 15 

~ ~o 

~ 5 

0 

~ -

...........~-=~. ~ . ~ . ~~ ......6~-~-~.~.~L-~-=.~ ..........~o... :~ ~ : ~ : : ~ : : ~:v v~:~ : ~ ........ ~....~ ........... ........... ~ ~...........,~ ...... ~.~ .......... .......... ........... .......... ~ ~ ~ ~ ...... 

~; .......... ~.......... ~........... ~..~........ ~.......... ~,..~...:............ ~.......... ~..... ............. 

........... ~ ........... , ........... • ........... . ............ 

• ~ 

......................................................... 

....................... ~........... ~........... ~.:......... . . 

................. .:...~ ........... ~....... :...)..~ ........ " 

: . 

~:. .......... ~ ........ 

: 

~; .............. '. .... 

; .......... ; .......... 

~ 

~....... ...~ ......... 

. . . 

~ ,I ~ 

a . ~ 

.~ 

• 

MALE MICE 

Vehicle Control (2 yr) 

Vehicle Control (3 yr) 

2,700 mg/kg (2 yr) 

2,700 ms/kg (3 yr) 

i i i ~ 

0 5 30 45 60 75 90 105 120 135 150 

WEEKS ON STUDY 

FIGURE 18 

Growth Curves for Male Mice Administered Salicylazosulfapyridine in Corn Oil by Gavage 

for 2 or 3 Years: 2-Year and 3-Year Restricted Feed Protocols 
Dietary Restriction, 1V~'P TR 460 77 

Pathology and Statistical Analyses 

This section describes the statistically significant or 

biologically noteworthy changes in the incidences of 

neoplasms or nonneoplastic lesions of the harderian 

gland, liver, lung, and spleen. Summaries of the 

incidences of neoplasms and nonneoplastic lesions and 

statistical analyses of primary neoplasms that occurred 

with an incidence of at least 5 % in at least one animal 

group are presented in Appendix F. 

Harderian Gland: At 3 years, the incidences of 

adenoma and adenoma or carcinoma (combined) were 

significantly less in dosed males than in the controls 

(Tables 24 and F2b). There were no significant 

differences in the incidence of harderian gland neoplasms between dosed and control males at 2 years. 

Liver: In contrast to the results for mice in the 

ad libitum feeding and the weight-matched controls 

protocols, the incidences of hepatocellular carcinoma 

and hepatocellular adenoma or carcinom a (combined) 

in the 2,700 mg/kg group fed a restricted diet were 

significantly less than those in the controls after 

2 years (Tables 24 and F2b). After 3 years, the 

incidence of hepatocellular carcinoma in dosed males 

was again significantly less than in the controls, but 

the combined incidences of hepatocellular adenoma or 

carcinoma in dosed and control males were similar. 

Lung: The incidences of alveolar/bronchiolar adenoma and alveolar/bronchiolar adenoma or carcinoma 

(combined) Were significantly less in dosed males than 

in the contrrols at 2 years, but not at 3 years 

(Tables 24 and F2b). However, at 3 years the incidence of alveolar/bronchiolar carcinoma in dosed 

males was significantly less than in the controls. 

Spleen: The incidence of hemosiderin pigmentation in 

dosed males was significantly greater than in the 

controls at 2 years (0/52, 39/50) and at 3 years (0/48, 

37/50) (Table Fab). 

TABLE 24 

Incidences of Selected Neoplasms in Male Mice ~n the Dietary Restriction Study 

of Salicylazosulfapyridine: 2.Year and 3-Year Restricted ~'eed ~'otocols 

Harderian Gland 

Adenoma 

Overall rate a 

Adjusted rate b 

Terminal rate c 

First incidence (days) d 

Logistic regression test 

Carcinoma 

Overall rate 

Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

(continued) 

2-Year'Restricted Feed 3-Year Restricted Feed 

Vehicle 2,700 mg/kg Vehicle 2,700 mg/kg 

Control Control 

2/52 (4%) 3/50 (6%) 6/48 (13%) 1/50 (2%) 

4.5% 8.6% 28.6% 2.8% 

1/42 (2%) 3/44 (7%) 5/20 (25 %) 0/34 (0%) 

660 730 (T) 1,079 1,076 

P=0.479 P=0.008N 

1/52 (2 %) 0/50 (0 %) 2/48 (4 %) 0/50 (0 %) 

3/52 (6%) 3/50 (6%) 8/48 (17%) 1/50 (2%) 

6.7% 8.6% 32.3% 2.8% 

1/42 (2%) 3/44 (7%) 5/20 (25 %) 0/34 (0%) 

660 730 (T) 750 1,076 

P=0.641 P=0.007N 
Dietary Restriction, NTP TR 460 

TABM~ 24 . • 

Incidences of Selected Neoplasms in Male Mice in the Dietary Restriction Study 

Of Salicylazosulfapyridine: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle 2,700 mg/kg Vehicle 2,700 mg/kg 

Control Control 

LiVer 

Hepatocellular Adenoma 

Overall rate 13/52 (25 % ) 9/50 (18 % ) 10/48 (21%) 14/50 (28%) 

Adjusted rate 34.6% 22.7% 35.0% 38.6% 

Terminal rate 10/42 (24%) 8/44. (18%) 5/20 (25 %) 12/34 (35 %) 

First incidence (days) ) .

Logistic regression test 

616 648 

P=0.274N 

555 933 

P--0.398 

Hepatocellular Carcinoma 

Overall rate 7/52 (13%) 1/50 (2%) 16/48 (33%) 6/50 (12%) 

Adjusted rate 14.9% 2.3% 42.2% 15.5% 

Terminal rate 4/42 (10%) 1/44 (2%) 3/20 (15%) 3/34 (9%) 

First incidence (days) 495 729 (T) 445 726 

Logistic regression test P=0.025N P=0.020N = 

Hepatocellular Adenoma or Carcinoma 

Overall rate 18/52 (35%) 9/50 (18%) 21/48 (44%) 18/50 (36%) 

Adjusted rate 42.2% 22.7% 55.5% 47.0% 

Terminal rate 12/42 (29%) 8/44 (18%) 6/20 (30%) 14/34 (41%). 

First incidence (days) 495 648 445 726 

Logistic regression test P=0.044N P=0~292N 

Lung 

Alveolar/bronchiolar Adenoma 

Overall rate 11/52 (21%) 3/50 (6%) 8/48 (17%) 7/50 (14%) 

Adjusted rate 53.2% 7.9% 33.1% 18.6% 

Terminal rate 9/42 (21%) 3/44 (7 %) 5/20 (25 %) 5/34 (15%) 

First incidence (days) 717 729 (T) 862 719 

Logistic regression test P=0.023N P=0.335N 

Alveolar/bronchiolar Carcinoma 

Overall rate 2/52 (4%) : 0/50 (0%) 13148 (27%) 6/50 (12%) 

Adjusted rate 13.0% 0.0% 38.4% 17.1% 

Terminal rate 1/42 (2%) 0/44 (0%) 3/20 (15%) 5/34 (15%) 

First incidence (days) 661 -~ 718 1,076 

Logistic regression test P=0.237N P=0.049N 

Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 13/52 (25~) 3/50 (6%) 18/48 (38%) 12/50 (24%) 

Adjusted rate 63.4% 7.9% 55.2% 31.8% 

Terminal rate 10/42 (24%) 3/44 (7%) 7/20 (35%) 9/34 (26%) 

First incidence (days) 661 729 (T) 718 719 

Logistic regression test P=0.009N P=0.075N 

(T)Terminal sacrifice 

a Number of animals with neoplasms per number of animals examined. Denominator is number of animals examined microscopically for 

liver and lung and number of animals necropsied for harderian gland. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence in animals surviving until the end of the study 

d In the dosed group columns are the P values corresponding to the pairwise comparisons between the controls and that dosed group. The 

logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in a dosed group is 

indicated by N. 

e Not applicable; no neoplasms in animal group 

c 
Dief~ry Restriction, NT~ TR ~ 79 

~COPOLAi~E ]~'~['I~D~O~OI~IIr[I~E ~]i¢..II~I-~D~.A.TE (~rA~rAG~ ~TILrD"Y 

,A~LL ~EED~NG ]PROTOCOLS 

Survival 

Estimates of 2-year Survival probabilities for male and 

female mice are shown in Tables 25 and 26 and in the 

Kaplan-Meier survival curves (Figures 19 and 20). 

The survival rates of all groups of dosed mice were 

similar to those of the respective controls; the survival 

rates of mice fed under different protocols were also 

generally similar. 

Weights 

The mean body weights of dosed male and female 

mice fed ad libitum were lower than those of the 

controls (Figure 21); at 1 year, mean body weights in 

the 25 mg/kg groups were 20% (males) and 23% 

(females) less than those of the respective controls 

(Table J9). The mean body weight of the dosed males 

was 94% that of the weight-matched controls; the 

mean body weight of dosed females was 99% that of 

the weight-matched controls (Table J9). During the 

second year of the study, the mean body weight of 

dosed males remained approximately 20% less than 

that of controls fed ad libitum and remained within 

10% that of the weight-matched controls; the mean 

body weight of females ranged from 16% to 25% less 

than that of the controls fed ad libitum and remained 

within 8 % that of the weight-matched controls. 

Among groups fed restricted diets, the mean body 

weights of males and females in the 25 mg/kg groups 

were slightly less than those of the respective controls 

at 1 year (Figure 22 and Table Jl0); dosed males confinued to weigh slightly less than the controls through 

the end of the study, but the mean body weights of 

dosed and control females were similar at the end of 

the study. 

Feed Consump~on, Clinical ~ndings, 

and Organ Weights 

Feed consumption by dosed and control mice fed 

ad libitum was similar (data not shown). Males and 

females administered scopolamine hydrobromide 

trihydrate had dilated pupils. 

For dosed males fed ad libitum, the absolute right 

kidney weight was significantly less and the relative 

right kidney weight was significantly greater than 

those of the controls fed ad libitum; dosed females fed 

ad libitum had a greater relative liver weight than the 

controls fed ad libitum(Tables I5a). Dosed males and 

females fed ad libitum had greater absolute and 

relative liver weights than the weight-matched controis. For dosed males fed a restricted diet, the 

absolute right kidney weight was less than that of the 

diet-restricted controls (Table I5b); there were no 

significant differences in organ weights between dosed 

and control females fed a restricted diet. 
80 Dietary Restriction, NTP TR 460 

'rA~LE 25 

Survival of Mice in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols . . 

. . 

Ad Libitum- Weight'Matched 25 mg/kg 

Fed Control Control 

Male 

Animals initially in study 70 60 70 

15-Month interim evaluation a 20 10 20 

Accidental deaths a 0 1 2 

Moribund 4 .2 7 

Natural deaths 6 6 2 

Animals surviving to study termination 40 41 39 

Percent probability of survival at the end of study b 81 85 83 

Mean survival (days) c 632 605 622 

Survival analysis d P= 1.000N ' 

Survival analysis e P= 1.000 

Female 

Animals initially in ,study 70 60 "70 

15-Month interim evaluation a 19 10 19 

Accidental deaths a 2 1 0 

Moribund 9 I0 7 

Natural deaths 7 3 6 

Animals surviving to study termination 33 36 38 

Percent probability of survival at the end of study 67 74 ~ 76 

Mean survival (days) 617 654 624 

Survival analysis P=0.656N 

Survival analysis P=I.000 

a Censored from analyses 

b Kaplan-Meier determinations 

c Mean of all deaths (uncensored, censored, and terminal sacrifice) 

d The results of the life table pairwise comparisons (Cox, 1972) with the controls fed ad libitum are in the eXl~osed group column. A lower 

mortality in the dosed group is indicated by N. 

e Result of life table pairwise comparison with the weight-matched controls 
Dietary Restriction, NT~ TR 460 8~ 

TABLE 2~ 

Survival of l~'Jlice in the Dietary Restriction Study 

2-Year and 3-~r ~tricted Feed ~rotoco~ 

of ScoponamineHydrobromide Trihydrate: 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle 25 mg/kg Vehicle 25 mg/kg 

Control Control 

Male 

Animals initially in study 60 60 50 50 

15-Month interim evaluation a 10 10 0 0 

Moribund 0 0 11 5 

Natural deaths 1 2 11 8 

Animals surviving to study termination 49 48 28 37 

Percent probability of survival at the end of study b 98 96 56 74 

Mean survival (days) e 674 663 1,004 1,027 

Survival analysis d P=0.986 P=0.130N 

Female 

Animals initially in study 60 60 50 50 

15-Month interim evaluation a 10 10 0 0 

Accidental deaths a 0 I 0 1 

Moribund 2 • 2 18 16 

Natural deaths 1 3 12 14 

Animals surviving to study termination 47 44 20 19 

Percent probability of survival at the end of study 94 90 40 39 

Mean survival (days) 670 660 951 945 

Survival analysis P = 0.691 P = 1.000N 

a Censored from analyses 

b Kaplan-Meier determinations 

c Mean of all deaths (uncensored, censored, and.terminal sacrifice) 

d The result of the life table pairwise comparison (Cox, 1972) with the controls is in the exposed group column. A lower mortality in the 

dosed group is indicated by N. 
82 

., 

Dietary Restriction, NTP TR 460 

1.00 - - ' 

~- o.~ ~ ......i .......... i .......... i ............. 

~ 0.90 D. 

.~ • 

~_ 0.85 

1~ 

~ 0.80 ~ ~c~ ................................................................ 

~ ~ Ad Libitum-Fed Control 

~ ~ Weight-Matched Control 

0.75 ~ 25 mglkg ......... : ...................................................... 

; 

: 

0.70 " ~ 

10 20 30 40 50 60 70 80 90 100 110 

WEEKS ON STUDY 

~.oo ~,~ 

0.90 

0.85 

0.80 

FEMALE MICE 

0.75 

0.70 

0.65 . 

10 20 30 40 50 60 70 80 90 100 110 

WEEKS ON STUDY 

F~6~ 19 

KaplanoMeier Survival Curves for Male and Female Mice 

Administered Scopolamine Hydrobromide Trihydrate in Water by Garage 

for 2 Years: Ad Libitum Feeding and Weight-Matched Controls Protocols 
D~eCa~'y ]Restriction, NTP TR ~60 83 

> 

1.0 

~ ~ ~ 

..................... i .......... ~ ............................. 

A i~ d ~ 

O 

0.6 

i .......... 

i ~ 

~ ~ 

........ :~... ~ 

,I ~ 

: : : 

Vehide Con~ol (2 yr) 

Vehide ~n~ol (3 yr) 

=~=~<=~> 

~'~'~' ~ "~ 

i i :... ~ i ! i .. i~ .... ........... ! ....... .... 

:: ~ ~ ~ ~ I~ 

~ ~ :: ~ ~ ~ 1 

............... ~ .......... :~ ......... ~ .......... :: ....... ~/... 

• ~- ' ~ 

0.5 :

I 

15 30 45 60 75 90 105 120 

~ I 

135 150 

WEEKS ON STUDY 

• 0.9 

~ 0.8-

1.1_ 

o 0.7 

~ 

~ 

m 

o 

~ 

~ 

0.6 

0.5 

0.4 

i[ .............. /-o- 

~ / ~ 

~ / ~ .......... ~'"'~ ~ 

~,c~ 

w~c0~,~0~ 

Vehicle Control (3 yr) 

~s =g~g (~ ~') 25mg~g(3yr) 

i 

~'---::~-d-'N~.~ 

? ~ ~ ~ ~ ~ 

~ ~ ....... ~.~ ............... ~ ~ ~ 

0.3 I = ~ = = 

15 30 45 60 75 90 105 120 135 150 

WEEKS ON STUDY 

~GURE 20 

Kap~an-Meier Survival Curves for Marie and ~ema~e IV~ice 

Achn~n~stered Scopo~a~nine Hydrobro~nide Trlhydrate ~n Water by Garage 

for 2 or 3 ~'ears: 2-fear Restricted ~eed and 3-great ]Restricted ~eed ]Pro¢oco~s 
Dietary Restriction, NTP TR 460 • ,, :~/ 

60 , ! i : 

................................................................................. • i............... i............... :i .... 

.- oOO::°•o~o i < 50 ................................ ~............... o.o.o--•--. ~ ............. ~ ............... -: ...... o-o---~--. ~I= i •• i i i i, 

Z --4~ ................................ ~-• ............................ i ........ [] ..... ~.~1..~ ........ :. ................ ~..... 

F- • i _o~ ~h^~91~^,~^ i 

:E 40-

:o " ~ A ~ ~ ~ ~z~ .~~ ...... ~{ ..... ~ '~~~~~ ..... ~a ~ ~ ~ ~ ~ ......... ~-~ ..... ~-:a--q--a .......... .-~ ................ ~............... ~............... ~................ :: ...... 35- >- . ~ , ~ ~ ~ ~ - ~ : 

O ~~~ ........ ~................ ~................ ~............... ~........ ~..... .~... .................. 30-

Z 

< 25- ~ ~ ~ [ MALE MICE ~ ~ " 

uJ ~ ~ • ; ~ ~ • Ad Libitu~Fed #ont,o, ~ ~.... ~ : : : : ~ Weight-Matched Control I ~ ............... , ................................. :......... ....... ,. ~ ~ =~ . . j'-'~'"-~[ 20-

I 15 I I,. I 

0 15 - 30 45 60 75 90 105 

WEEKS ON STUDY , . 

60 

55 ............................................................................ • .... • ...... i i i o ~° i °O~o i 

< 50 ................ i ............... i ............... • • • ~ ~............... ! .............................. : :! ...... • ....... i .... • [ 

! o • ! ! : z 45 ................ : ............... < ................................ ~............................... -: ................ ~ .... 

i i ° ~i . ~i~::~Ba~ ~ :: :3: 

40 ................ ;i ................................................... o o° &#ee~Ba i ................. ~aa~~a ..... 

~ • , 

35 ................ ~o .......... -5-~,.~-a ......................... ~........................................... f .... 

~:: ~ a~ • :: 

0 ~0 ..... ~-~ ...... ~---~ ................ ~ ............... ~............... : ............... ~............ ~--.~.-.-~ 

Z ~= = ~ ~ ~ ~'~ I ~ I 25- ~ ............. ~................ ::................ ~.... • ~U~,~-~.~o,~ro~ I-i ..... I 

~ ...... ... ~ :: :: :: a Weight Matched Con~ol ~ 

20- " ................................................................. ~ ~.~. I~[ 

I 15 ~ ~ ~ 

0 15 30 45 60 75 90 105 

WEEKS ON STUDY 

FXGURE 21 

Growth Curves for Male and Female Mice Administered 

Scopolamine Hydrobromide Trihydrate in Water by. Gavage for 2 Years: 

Ad Libitum Feeding and Weight-Matc.hed Controls Protocols . 

. , 
Dietars, Restriction, NTP T~ 4~ 8S 

45 / . 

40 .t .......... i......... :i .......... i........... :2~;~i~_ ........ i.......... i.......... i.......... :i ..... ............. 

Z . i i i ooiool o i i i ooi i 

I i ! i i i :o i o o o~ ! 

35 I .......... i .......... i ....... 62o~°~~~! ~8~-o ...... i ........... i~-b-!2 

[ ! [] i ~0 ! ~ ~ m ~ ~ ~ 0 0 u LiJ ~ - ~ o~ =O ~ ~ ~ - ~ " ~ ~ ~I ~ 4~g~&~ ~ ~a~.aeTvvvyvvv~vv ~v ~u q .......... ~g-e~-~-~---~ ......... ~ ......... ~ ..... v-~---v,-,-~ .......... ~.......... ~.......... : ....... 

O / ~0~ ::~ :: :' ~ ' " " ~ 

I ~--~ ' ~ ~ ~ ~ . MACE MICE ~ 

<

Z 25 ~ " ~ ~ ~ ~ I o w.,~,~oo.,.o,{=~0 I == I,LI ~'{ ................................. ~........... : ........... ~'"l ~ Vehicle Control (3yr) I'":: ...... 

" I~ I ~ =~ ~me {= y0 I ~ 

2o ~ I,,?, ~T~">~, , I~ 

0 15 30 45 60 75 90 105 120 135 150 

WEEKS ON STUDY 

45 

40 .............................................. : ........................................................................... 

z 3 5 q .......... ......... ............ ........... ........ .................. 

! ~o~o~ ~ i ~ i : | , , LU 30 4 { .......... ...... 8~-m-m-°-: ~- -~ ......... 9~A~m~,. .......~ °o:; ...... o~ .......

] :: ~XT~ ~ ~ ~ - ~ ~ ~ : : ~ ~ e - ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ 

O ........ ~.~....~ .......... ~...................... ~ .......... ~... :: ~ ~ .......... 

Z ~5~ ~ :: ~ ~ :: 1 ~,~e 1 ~ 

LU ~ ~ ~ ;: :: ~ ~ / o V~h,~,~ co~t,o~ (~ ~0 I ~ 

M~ ~ ~ ~ ~ :: ~ / ~ V~h~ Co.t,o~{s ~r~ I :: 

20 - ~ ................................................................. r 1 ~ ~s=~r~ • I'"~ ....... 

/ ~ ~,~,,r~ 1~ : : : : 

I I I I 

0 . 15 30 45 60 75 90 I05 120 135 150 

WEEKS ON STUDY 

FIGURE 22 

Growth Curves for Male and Female ]V~ce Administered 

S¢opo~an~ine Hy~irobro~nide Tr~hydrate in Water by Gavage for 2 or 3 ~ears: 

2-~fearr Restricted Feed and 3-Year Restricted Feed Protocols 
86 Dietary Restriction,-NTP TR 460 

Pathology and Statistical Analyses 

This section describes the statistically significant or 

biologically noteworthy changes in the incidences of 

n.eoplas, m ~ and nonneoplastic lesions of the liver. 

Summaries of the incidences of neoplasms and nonneoplastic lesions and statistical analyses of primary 

neoplasms that occurred with an incidence of at least 

5% in at least one animal group, are presented in 

Appendix G for male mice and Appendix H for 

female mice. Pathologic descriptions of neoplasms 

and nonneoplastic lesions occurring in rats in the 

scopolamine hydrobromide trihydrate study are 

provided in NTP Technical Report 445 (NTP, 1997a). 

No significantly increased neoplasm incidences were 

noted in dosed mice regardless of the method of 

feeding; Suggesting that scopolamine hydrobromide 

trihydrate is not carcinogenic in mice. The incidences 

of liver neoplasms in dosed mice fed ad libitum 

(Table 27) and of a variety of nonneoplastic lesions in 

the weight-matched controls, diet-restricted controls, 

and mice in the 25 mg/kg groups fed ad libitum or 

receiving restricted diets were less than the incidences 

of these lesions in controls fed ad libitum, suggesting 

that the lower incidences were related to body weight 

depression, 
Die~ Res~cfion, ~ TS ~6~ ~7 

TA~L~ 27 

~c~dlences o~ Heop~asnas ~n~ Honneo~mfic Lesions o~ ~he Liver ~n Mice 

~ the D~e~ R~r~cfion Study o~ Sco~ne Hy~robro~e ~hy~te: 

Ad L~m ~ee~ng and Weight-Matched Control ~ot~o~ 

Ad Libitum- 

~'ed Control 

Weight-Matched 

Control 

25 mg/~q~ 

MaRe 

~$-Month Interim Evaluation 

Number Examined Microscopically 

Hepatocellular Adenoma a 

Hepatoeellular Carcinoma 

10 

2 

0 

10 

1 

0 

10 

0 

1 

2-Yea~Study 

Number Examined Microscopically 

Basophilic Focus 

Clear Cell Focus 

Eosinophilic Focus 

Mixed Cell Focus 

50 

3 

12 

21 

5 

50 

0 

2 

0 

2 

50 

0 

0"* 

2** 

1 

Hepatocellular Adenoma 

Overall rate b 

Adjusted rate c 

Terminal rate d 

First incidence (days) 

Logistic regression test e 

Logistic regression test f 

26/50 (52%) 

59.0% 

22/40 (55%) 

680 

5/50 (10%) 

11.9% 

4/41 (10%) 

721 

8/50 (16%) 

19.1% 

6/39 (15%) 

587 

P<O.OOIN 

P=0.308 

Hepatocellular Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

6/50 (12%) 

14.2% 

4140 (10%) 

700 

5/50 (10%) 

11.8% 

4/41 (10%) 

532 

7/50 (14%) 

17.3% 

6/39 (15%) 

622 

P=0o468 

P=0.413 

Hepatocellular Adenoma or Carcinoma g 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

30/50 (60%) 

65.2% 

24/40 (60%) 

680 

10/50 (20%) 

23.2% 

8/41 (20%) 

532 

15/50 (30%) 

35.3% 

12/39 (31%) 

587 

Pffi0.004N 

Pffi0.217 

(continued) 
88 Dietary Restriction, NTP TR 460 

TABLE 27 

Incidences of Neoplasms and Nonneoplastic Lesions of the Liver in Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 25 mg/kg ,

Fed Control Control 

Female 

15-Month Interim Evaluation 

Number Examined Microscopically 

Hepatoceilular Adenoma 

2-Year Study 

Number Examined Microscopically 

Clear Cell Focus 

Eosinophilic Focus 

Mixed Cell Focus 

HePatocellular Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (~lays) 

Logistic regression test 

Logistic regression test 

Hepatocellular Carcinoma 

• Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

Hepatocellular Adenoma Or Carcinoma h 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

Logistic regression test 

.10 

1 

51 

0 

17 

6 

15/51 (29%) 

42.3% 

13/33 (39%) 

604 

8/51 (16%) 

21.1% 

5/33 (15%) 

594 

22/51 (43 %) 

57.1% 

17/33 (52%) 

594 

10 I0 

0 0 

50 51 

1 0 

4 9* 

1 3 

7/50 (14%) 6/51 (12%) 

18.9% 15.4% 

6/36 (17%) . ~5/38 (13%) 

721 694 ,. 

P=0.017N 

P=0.526N 

2/50 (4%) 4/51 i8%) 

4.8% 10.3% 

1/36 (3%) 3/38 (8%) 

611 694 

P=0.170N 

P=0.339 

9/50 (18%) 9/51 (18%) 

23.2 % 23.1% 

7/36 (19%) 8/38 (21%! 

611 694 

P=0.003N 

P=0.578 

* Significantly different (P~0.05) from the adlibiturn-fed control group by the logistic regressioh test 

** P~0.01 

a Number of animals with lesion 

b: Number of animals with neoplasms per number of animals with liver examined microscopi~cally 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

d Observed incidence in animalssurviving until the end of the study .... 

e In the dosed group column are the P values corresponding to the pairwise comparisons between the ad libitum-fed controls and the dosed 

group. The logistic regression test regards neoplasms in animals dying prior to terminal kill as nonfatal. A lower incidence in the dosed 

group is indicated by N. 

f Result of pairwise comparison between the weight-matched controls and the dosed group 

g I-~istorical incidence for 2-year NTP studies with control groups receiving water by gavage and fed ad libitum (mean ± standard deviation): 

74/315 (23.5% ± 7.2%); range, 14%-36% 

h Historical incidence: 21/315 (6.7% ± 4.2%); range, 2%-12% 

c 
89 !~ 

[ D I$C J$$ I©M AND. C©NCLU$]I©N$ 

The studies presented here were undertaken to compare outcomes, when four chemicals were evaluated 

under typical NTP bioassay conditions and by protocols employing dietary restriction. Specifically, 

experiments were designed to evaluate the effect of a 

15 % body weight reduction (attained through feed 

restriction) on the sensitivity of the bioassay to detect 

• carcinogenicity compared with the results from 

concurrent evaluations with conventional ad libitumfed animals. Because animals fed restricted diets live 

longer than those fed ad libitum, an arbitrary evaluation at 2 years.results in comparisons at disproportionate times .in the respective lifespans; thereby 

potentially masking age-dependent effects. Thus, an 

additional 3-year/20% .survival study was conducted 

to determine whether an additional year of exposure 

to the chemicals would influence the neoplasm profile 

of animals fed restricted diets. Finally, because body 

weight changes alone, without chemical exposure, 

alter the expression of a variety of lesions, a compari= 

son employing a weight-matched (pair-weighted) 

control group was included. 

Overall, feed restriction increased the survival of rats 

and, to a lesser extent, mice, although not consistently 

in all studies. The enhanced longevity and decreased 

incidences of neoplasms and nonneoplastic lesions 

provide evidence that the restricted diets were nutritionally adequate for long-term health and survival. 

Thus, conditions of "undernutrition without malnutrition" (Weindruch, 1984) were achieved. The 

weight-matching procedur e was generally effective in 

producing a control group with body weights similar 

to those of the dosed animals. Salicylazosulfapyridine 

had such a minor effect on the body weight of male 

rats that weight matching was not attempted, thereby 

resulting in a second ad libitum-fed control group. It 

was initially suspected that feed restriction of grouphoused rats might result ina heterogenous population 

of animals with feed consumption ranging from 

minimally restricted to severely restricted. Because 

the group housing of rats precluded precise 

assessment of individual animal feed consumption, 

variability in body weights was employed as a surrogate indicator of variability in individual feed 

consumption. Examination of the standard deviations 

of the weekly body weights of ad libitum-fed versus 

feed-restricted control animals showed that this 

variability was not greater in feed-restricted animals 

(data not shown). In fact, the converse was observed; 

body weights of the feed-restricted animals were 

generally more homogenous. 

The results presented in this report were consistent 

with observations by others that dietary restriction 

increases the survival rates and decreases the incidences of neoplasms and nonneoplastic lesions at a 

variety of sites in control and dosed animals. 

Table 28 provides a compilation of the increases and 

decreases in neoplasm responses to the four chemicals 

under the various protocols. The organ sites listed are 

those at which any protocol yielded a statistically 

significant increase or decrease. Following is a 

discussion of the differences in response between the 

ad libitum feeding, weight-matched, and restricted 

feed protocols for each chemical. 

~uty~ Ben~y~ ~htha~ate 

The incidences of pancreatic acinar cell neoplasms 

were greater in male rats receiving butyl benzyl 

phthalate in feed than in the controls fed ad libitum or 

the weight-matched controls (Table 28). A concomitant increase in the incidence of pancreatic acinar cell 

hyperplasia (ad libitum-fed controls, 8%; weightmatched controls, 4%; 12,000 ppm, 24%) suggests 

that these lesions were related to chemical administration. This interpretation is Supported by the bbservation that some other peroxisome proliferators also 

cause pancreatic acinar cell neoplasms (see NTP, 

1997a). The incidences of these neoplasms were not 

increased in rats fed a restricted diet for 2 years. 

However, there was some suggestio n of a treatmentrelated increase in the incidence of proliferative 

lesions of the pancreatic acinar cell after a longer 

period of chemical exposure: in the 30-month 

restricted feed protocol, three exposed male rats 
90 Dietary Restriction,NTP TR 460 

TAntJ~ 28 

Summaryof Neoplasm Incidences in Long-Term Bioassays by Various Feeding Protocols a 

Control vs. Dosed Group 

Chemical/ Sex/ Neoplasm Protocol 1 Protocol 2 Protocol 3 Protocol 4 

Route Species Site Ad Libitum Weight-Matched Restricted Feed Restricted Feed 

Feeding Controls (2 years) (up to 3 years) 

Increased Incidences 

Butyl benzyl phthalate Male rats : Leukemia 62% vs. 60% ' 30% vs. 60%** 42% vs. 54% 78% ~s. 72% 

(Feed) Adrenal medulla 20% vs. 20% 6% vs. 20%* 8% vs. 10% 18% vs. 12% 

Pancreas 6% vs. 22%* 2% .vs. 22%** 0% vs. 0% 0% vs. 6% 

Female rats Leukemia 42% vs. 38% 26% vs. 38%* 32% vs. 36% 58% vs. 78% 

Urinary bladder • 2%vs. 4% ~ 0% vs. 4% - 0% vs. 4% 2% vs. 12% 

t-Butylhydroquinone Male rats Preputial,gland " 12% vs., 13% 0% vs. 13%** NA 3% vs. 5% ' 

(Feed) Female rats Clitoral gland 21% vs. 23% 10% :vs. 23%* NA 8% vs. 25%* 

Salicylazosulfapyridine Male rats Urinary bladder 0% vs. 12%* 0% vs. 12%* ' 0% vs. 0% 0% vs. 2% 

(Corn oil garage) Male mi,ce Liver 48% vs: 88%** 28% vs. 88%** 35% vs. 18%* 44% vs. 36% 

Scopolamine hydrobromide Male and 

trihydrate (Water garage) female mice No carcinogenic effects under any protocol 

Decreased Incidences " 

Butyl benzyl phthalate Male rats " Preputial gland 10% vs. 0%* 12% vs. 0%* 2% vs. 2% 2% vs. 4% 

(Feed) Female rats Clitoral gland 14% vs. 8% 6% vs. 8% 6% vs. 6% 20% vs. 4%* 

Mammary gland 58% vs. 22%** 14% vs. 22% 26% vs. 4%** 48% vs. 12%** 

Pituitary gland 45% vs. 26% 15% vs. 26% , 30% vs. 12%* 52% vs. 32%* 

Skin (subcut.) 2% vs. 0% 2% vs. 0% 2% vs. 0% 8% vS. 0%* 

t-Butylhydroquinone Male rats Pancreas 5% vs. 0%* 0~ vs. 0% NA 0% vs. 0% 

(F.eed) ' ' ' Pituitary gland 32% vs. 10%** 32% vs. 10%** NA 23% vs. 29% 

Female rats Adrenal cortex ~/% vs.0%* 0% vs. 0% NA 0% vs. 2% 

Mammary gland 80% vs. 50%** 43% vs, 50% NA 52% vs. 28%** 

Pituitary gland 43% vs. 47% 53% vs. 47% NA 51% vs. 30%** 

Salicylaz~sul fapyridine Male rats Adrenal medulla 32% vs. 16% 20% vs. 16% 18%vs. 2%** 31% vs. 16%** 

(Corn oil gavage) Leukemia 26% vs. 6%* 20% vs. 6% 22% vs. 4%** 49% vs. 16%** 

Lung 4% vs. 0% 2% vs. 0% 4% vs. 0% 6% vs. 0%* 

Male mice Liver 48% vs. 88% 28% vs. 88% 35% vs. 18%* 44% vs. 36% 

Forestomach 6% vs. 0% 10% vs. 0%* 4% vs. 4% 4% vs. 4% 

Harderian gland 6% vs..4% 4% vs. 4% '6% vs. 6% 17% vs.-2%** 

Lung 28% vs. 22% 18% vs.'22% 25% vs. 6%** 38% vs. 24% 

Scopolamine hydrobrornide Male mice Liver 60% vs. 30%** 20% vs. 30% 10% vs. 2% 26% vs. 26% 

trihydram(Water gavage) Female mice Liver 43% vs. 18%** 18% vs. 18% 6% vs. 6% 30% vs. 22% 

* Significantly different (P~0.05) from the control group by !ife table analysis (leukemia only) or logistic regression analysis 

** P~0.01 

a Rats in the t-butylhydroqninone study were fed ad libitum or received restricted diets for 30 months or until survival was reduced to 20% 

in the control or.exposed group; males in the ad libitum feeding and weight-matched groups received 28 months of exposure, and all other 

groups received 30 months of. exposure. In the other studies, animals in the first, second, and third protocols received 24 months of 

exposure; under the fburth protocol, animals received restricted diets for 36 months or until survival was reduced to 20%. In the butyl 

benzyl phthalate study, males received 30 months and females 32 monthg of exposure. In the salicylazosulfapyridine studies, male rats 

received 30 months and male mice 36 months of dosing. Mice in the scgpo!amine hydrobromide trihydrate study received 36 months of 

dosing. 
i 

D~e~ry ~es~r~cfion, NT~ TS 460 

developed pancreatic acinar cell adenomas. Three 

exposed male rats in the 2-year feed-restricted group 

and two in the 30-month feed-restricted group had 

acinar cell hyperplasia; no pancreatic hyperplasia or 

adenomas were observed in either control group. 

There is evidence from the "studies in this report and 

from the literature that lower body weights are associated with a decrease in the incidence of pancreatic 

acinar cell adenoma in rats. For example, the incidence of pancreatic acinar cell adenoma was lower in 

feed-restricted male control rats than in the ad libitumfed controls in the three rat studies presented in this 

report: butyl benzyl phthalate study, 0% versus 6%; 

t-butylhydroquinone study, 0 % versus 5 %; salicylazosulfapyridine study, 10% versus 24% (P<0.05). The 

consistency of the results for these studies indicates 

that the stronger chemical-related proliferative 

response in the ad libitum feeding protocol compared 

to the restricted feed protocol is biologically significant. The low incidences of proliferative lesions in 

the dosed rats fed restricted diets for up to 30 months 

suggest that the effects of feed restriction may have 

delayed the development of spontaneous and 

treatment-related lesions of the pancreas. 

Experimentally induced carcinogenesis of the exocrine 

pancreas is sensitive to dietary factors such as fat 

quantity and quality (Roebuck, 1986; NTP, 1994a) 

and can be inhibited by several different restriction 

regimens, including feed restriction (Roebuck et al., 

1981), caloric restriction, and meal feeding (Roebuck 

et al., 1993). The reduction of feed intake by meal 

feeding for 5 hours per day for only the last 2 months 

of a 14-month study resulted in marked reduction of 

azaserine-induced pancreatic neoplasms without 

altering the mean body weights of these restricted 

animals relative to the ad libitum-fed controls 

(Roebuck et al., 1993). Pancreatic acinar cells 

isolated from ad libitum-fed F334 rats exhibited faster 

growth rates in vitro and a greater proclivity for 

malignant transformation than those isolated from 

restricted rats (Hass et al., 1992). 

The incidence of adrenal gland pheochromocytoma 

was marginally greater in exposed male rats than in 

the weight-matched controls (P=0.028; Table A2a). 

However, the dosed group incidence was not 

91 

significantly different from that in the ad libitum-fed 

controls, and the incidence of pheochromocytoma was 

not increased in exposed males in either of the 

restricted-feed protocols. Thus, this statistically significant difference reflects the decreased incidence of 

adrenal pheochromocytoma in the weight-matched 

controls relative to the ad libitum-fed controls. 

The occurrence of adrenal gland pheochromocytoma 

is not strongly correlated with body weight in control 

animals (Seilkop, 1995), so the reduced neoplasm ? 

incidence in weight-matched controls (6 %) relative to 

the ad libitum-fed controls (20%) was not expected. 

In the methylphenidate hydroc~loride study (NTP, 

1995), the maximum mean body weight of 430 g 

achieved by untreated male control rats approximates 

the mean body weight of 438 g observed foi~ the 

weight-matched controls in the butyl benzyl phthalate 

study. The incidence of adrenal gland pheochromocytoma in untreated control male rats in ,that study 

(37%) was greater than the incidence of 20% 

observed in ad libitum-fed, exposed male rats in the 

butyl benzyl phthalate study and was much greater 

than the incidence of 6% in the equivalently 

weight-matched controls in the butyl benzyl phthalate 

study. The overall incidence of adrenal gland pheochromocytoma in untreated controls in the current 

NTP historical control database is 33.7 % (398/1,182), 

considerably greater than the 20% observed in the 

exposed group. For these reasons, the greater incidence of adrenal medulla pheochromocytoma in 

exposed male rats relative to the weight-matched 

controls is considered unlikely to be a chemicalrelated effect. 

Urinary bladder transitional cell neoplasms are rarely 

observed in untreated female F344/N rats and 

occurred in only 4 of 1,182 (0.3%) control female rats 

in the current NTP historical database. The four 

incidences in the historical, control database were all 

papillomas; no carcinomas were observed. In the 

32-month restricted feed protocol of the butyl benzyl 

phthalate study, urinary bladder transitional cell 

neoplasms occurred in six exposed female rats (two 

papillomas and four carcinomas); one papilloma 

occurred in the controls. In the 2-year restricted feed 

protocol, two exposed females and no control females 

had urinary bladder papillomas. Urinary bladder 
92 

papillomas als0 Occurred in two exposed females fed 

ad libitum and in one ad libitum-fed control fe.ma!e; 

none occurred in the weight-matched controls. .. . 

• 

The increasedAncidences of these Urinary. bladder 

neoplasms were accompanie d by increased incidences 

of urinary bladder transitional epithelium hyperplasia 

in exposed females in the ad libitum feeding protocol (2 % vs. 20 %), the weight-matched controls protocol (0% vs. 20%), the 2-year restricted feed protocol 

(0% vs. 28%) and the 32-month restricted feed 

prbtocol (0% vs. 32%). The consistency of the 

urinary bladder neoplasm/hyperplasia response 

indicates that it is related to chemical administration. 

This effect was only observed in feed-restricted 

females, which had greater survival rates at 2 years 

and which were administered butyl benzyl phthalate 

for a longer period of time than ad libitum-fed rats. 

It is not certain if there wasa sex difference in 

response to butyl benzyl phthalate. In the feed, 

restricted groups, one exposed male at 2 years and 

two at 30 months had urinary bladder neoplasms. 

However, exposed males received a concentration O f 

. butyl benzyl phthalate that was half that administered 

to females. . 

The incidence of mononuclear cell leukemia was 

greater in exposed male and female rats fedad libitum 

thanin the weight-matched controls (Table 28). 

However, these increases were not significantly 

different from those in ~he ad libitum-fed controls, and 

no treatment-related increases in the incidence Of 

mononuclear cell leukemia occurred in either of the 

restricted feed protocols. Thus, these statistically 

significant differences were considered due to the 

decreased incidences of mononuclear cell leukemia in 

the weight-matched controls (males, 30%; females., 

26 %) relative to ad libitum-fed controls (males, 62%; 

females, 42%). .~ 

These increased incidences of mononuclear cell 

leukemia are considered an incidental finding for 

several reasons. First, these increases were only 

Observed in the weight-matched controls protocol. 

Also, although the increased incidence in exposed 

male rats (30% vs. 60%) was highly significant, ~he 

increased incidence in females was not as greai (26 % 

vs. 38%) and was significant by the life table test 

(P=0.034) but not by logistic regression analysis 

(P=0.302). 

:. Die~ry Restriction, NTP TR 460 

The weight,matched Contr01s ~ in the ~ butyl benzyl 

phthalate study were the only groups in the dietary 

restriction studies with both reduced mean body 

weights, anddecreased :incidences of mononuclear cell 

leukemia: In the t-butylhydroquinone study, for 

example, the weight-matched and feed-restricted 

Control s weighed much less than the ad libitum-fed 

controls and yet did not have lower incidences of 

mononuclear cell leukemia after 28 or 30 months 

(males: ad libitum-fed controls, 65%; weightmatched controls, 75 %; feed-restricted controls, 77 %; 

females: 45%, 42%, 62%). Similarly, in male rats 

in the salicylazosulfapyridine study, the ad libitum-fed 

and. weight-matched controls weighed much more than 

the feed-restricted controls, and yet the mononuclear 

cell •leukemia rates were similar among the three 

groups (26%, 20%, and 22%, respectively). Finally~ 

even in the butyl benzyl phthalate study, the inci- .~ 

dence., of mononuclear cell leukemia in the feedrestrictedc0ntrol~male tats (42%) was greater than 

that in the weight-matched controls (30%), even 

though the feed-restricted controls weighed much less 

than the weight=matched controls• (368 g vs. 400 g at 

w6ek~53) ..... , . , : 

The incidence ~ of mononuclear cell leukemia is w6akly 

and inconsistently co.rrelated with body weight in 

• control animals (Seilk0p, 1995). In the previously 

mentioned • : methylphenidate hydrochloride study 

(NTP, 1995), in. which the maximum mean body 

weight ..achieved by untreated male control rats 

approximates' that observed in the weight-matched 

control males in the butyl benzyl phthalate study, the 

• incidence oJY mononuclear cell !euk6mia in the 

untreated control group was 58 %; this incidence .is 

similar..to that observed in the ad libitum-fed males 

.administered.butyl. benzyl phthalate and much greater 

than the incidence in the weight-matched controls in 

the butyl .benzyl phthalate study. 

in contrast, other investigators have reported thatfeed 

~restriction.delaysthe occurrence (Yu et al., 1982) and 

lessens the severity (Maeda et al., 1985)of leukemia 

in F344 rats. Feed-restricted rats in lifetime studies 

have been shown to have greater incidences of monbnuclear cell leukemia than rats fed ad libitum, probably because the feed-restricted rats lived longer 

(Shimokawa et al., 1993; Thurman et al., 1994). 

Feed restriction decreases the rate of transplanted 

mononuclear cell leukemia progression in Fischer 344 

. • 
Dietary Restriction, NTP TR 460 93 • . 

rats through its influence on cell proliferation .via 

suppression of the GH:IGF-1 axis and its enhance: 

ment of host defenses against neoplasti~ Cells 

(Hursting et al., 1993). Finally; mononuclear ceil 

leukemia is known to be sensitive to nut~itionai 

factors; the incidence of this neoplasm in male rats, 

but not in females, "is decreased by g~vag e administration of corn oil (HaSema net al., 1985; Hursting 

et al., !994). ~ 

In a previous NTP study of butyl berizyl phthalate 

(NTP, 1982a), female rats i~eceiving .the highest 

concentration (12,000 ppm) had a greater incidence of 

mononuclear Cell leukemia than the controls. However, this concentration did not cause an increased 

incidence of mononuclear cell' leukemia in the more 

recent NTP study (NTP, 1997c), andthe increased 

incidence observed in the current study at an even 

higher concentration (24,000 ppm) was only marginally significant, as noted above. Thus, this effect 

is considered to be an uncertain finding. 

t-Bu~ylhydroqu~nor~e .. 

The three protocols for the study of t:butylhydro~ 

quinone had disparate incidences of clitoral and ,. 

preputial gland neoplasms in rats. No increases in the 

incidences of these neoplasms w~ere Observed in 

exposed rats in the ad libitum feeding protocol 

(Table 28). However, the incidences of these neoplasms in the weight-matched controls were markedly 

less than those in the ad libitum-fed controls; as a 

result, the incidences in exposed males (13%) an d 

female rats (23 %) were significantly greater (P < 0.01) 

than those in the weight-matched controls (males, 0 %; 

females, 10%).. Exposed female rats fed a restricted 

diet also had a significantly greater incidence of , 

clitoral gland neoplasms after 30 months than the 

feed-restricted controls (8 % Vs. 25 %; P < 0.05). 

The increases in these neoplasm incidences are 

consideredan uncertain finding for several reasons. 

First, the incidence s of clitoral and preputial gland • 

neoplasm s are not strongly correlated with body 

weight in control animals (Seilkop, 1995), so the 

reduced neoplasm incidence in the weight-matched 

controls was not expected. Second, the clitoral and 

preputial gland neoplasm responses among the d!etary 

restriction studies were inconsistent. For example, 

the weight-matched control male rats: in the-butyl 

benzyl phthalate study had approximately the same. 

mean body weight as the weight-matched controls in 

the t-butylhydroquinone study and yet had a much 

greater incidence of preputial gland neoplasms at 

2 years (12%) than the incidence in the controls in the 

t-butylhYdroquinone study at 28 months (0%). t 

Similarly, the incidence of clitoral gland neoplasms in 

feed-restricted female control rats at 30 months in the 

t,butylhydroquinone study (8%) is much lower than 

the corresponding control incidence in the butyl 

benzyl phthalate study (20 %; Table 28). These comparisons suggest that control incidences of clitoral and 

preputial gland neoplasms in the t-butylhydroquinone 

study may have been unusually low and that the 

greater neoplasm incidences in the exposed groups are 

therefore an incidental finding. 

Sa~icy~a~osulfapyr~dine 

Rats: Urinary bladder neoplasms were observed in 

six male rats receiving salicylazosulfapyridine by 

gavage but not in the ad libitum-fed or weightmatched controls (Table 28). The incidence of 

urinary bladder hyperplasia was concomitantly 

increased (ad libitum-fed controls, 0%; weightmatched controls, 0%; 337.5 mg/kg, 82%). The 

Urinary bladder neoplasms all occurred in rats grossly 

observed at necropsy to have urinary bladder calculi; 

in addition, urinary bladder concretions were 

observed microscopically in 10 of 50 dosed rats fed 

ad libitum but in neither control group. 

There was little evidence of urinary bladder concretion in feed-restricted rats, with only one dosed male 

affected after 30 months. The incidence of urinary 

bladder.hyperplasia in dosed, feed-restricted rats was 

still greater than that in the controls at 2 years (0 % vs. 

14%) and at 30 months (0% vs. 16%), but the magnitude of the response was much less than that 

observed in the ad libitum feeding and weightmatched controls protocols. A single dosed rat in the 

30-month feed restriction protocol had a urinary 

bladder neoplasm (one other rat had urinary bladder 

concretion). Thus, it appears that the absence of 

urinary bladder neoplasms in dosed rats in the 

restricted feed groups may have been related to the 

absence of urinary bladder calculi and concretions in 

these groups. 
94 Dietary Restriction, NTP TR 460 

Feed restriction has been shown to delay the onset and 

slow the age-related severity of nephropathy in F344 

rats (Yu et al., 1982). The beneficial effects of both 

calorie and protein reduction on the severity of 

nephropathy has been observed in F344 rats and in 

other rat strains (Saxton and Kimball, 1941; Bras and 

Ross, 1964; Tucker et al., 1976; Everitt et al., 1982). 

The consumption of less food (and less protein) may 

have resulted in the decreased severity of nephropathy 

in diet-restricted rats, contributing to a more effective 

• removal of precipitated salicylazosulfapyridine or its 

metabolites from the kidney and urinary bladder and 

decreasing the formation of crystals and subsequent 

urinary tract concretions. 

The incidences of mononuclear cell leukemia in dosed 

male rats in the ad libitum feeding protocol and the 

2-year and 30-month restricted feed protocols were 

significantly less than those in the respective controls. 

The incidences of hematopoietic cell proliferation and 

hemosiderin pigmentation in the spleen were concurrently increased in dosed males. The biological 

significance of these changes is uncertain; in a number 

of NTP studies, chemicals i causing significantly 

decreased incidences of mononuclear cell leukemia 

have also caused changes suggestive of hematopoietic 

toxicity at 13 weeks, including increased incidences of 

splenic hematopoietic cell proliferation, • hemosiderin 

pigmentation, and fibrosis (NTP, 1982b, 1989, 

1994b, 1996). However, this association did not 

occur in ' all studies in which the incidence of mononuclear cell leukemia was decreased in treated groups 

(NTP, 1987, 1992). The consistency of the findings 

in the current studies suggest that salicylazosulfapyridine administration may indeed cause decreased 

incidences of mononuclear cell leukemia in rats. 

Mice:' Dosed mice fed ad libitum had increased 

incidences of hepatocelluiar adenoma relative to the 

ad libitum-fed and weight-matched controls 

(Table 28). Additionally, increased incidences of 

liver neoplasms also occurred in dosed female mice 

and in male mice receiving lower doses of salicylazosulfapyridine that caused no body weight changes 

(NTP, 1997b). No liver neoplasm response was 

observed in the restricted feed protocols, even after 

3 years of dosing. In fact, a significantly lower 

incidence of liver neoplasms was observed in dosed 

mice fed a restricted diet (Table 28). The difference 

in liver neoplasm incidences between ad libitum-fed, 

dosed male mice (88%) and feed-restricted, dosed 

male mice (18%) was marked. Incidences of liver 

neoplasms in B6C3F~ mice are strongly correlated 

with body weight (Turturro et al., 1993; Seilkop, 

1995). 

Because .the incidences of liver neoplasms were 

decreased in the control and dosed feed-restricted 

mice, it appears that the decreased incidences of these 

neoplasms were attributable in part to the lower mean 

body weights that resulted from feed restriction and 

•from chemical treatment. In this case, factors associated with body weight, feed intake, or both were 

clearly stronger inhibitors of mouse liver carcinogenesis than the positive carcinogenic effects of 

salicylazosulfapyridine. It should be noted that the 

•maximum tolerated dose of 2,700 mg/kg was used in 

these studies, and the lower mean body weight of 

dosed male mice fed a restricted diet (85 % of the 

ad libitum-fed control mean body weight) may have 

contributed to the decreased incidence of liver neoplasms- in this group. 

Under conditions that significantly decreased the 

spontaneous formation of liver neoplasms in feedrestricted B6C3Fi mice in comparison to ad libitumfed mice, the apoptosis:proliferation ratio for. 

hepatocytes was enhancedin the feed-restricted ~ice; 

this change is consistent with the observed protective 

effects of feed restriction against cancer (James and 

Muskhelishvili, 1994). Identical conclusions were 

reached in studies of chemical-induced hyperplasia 

and neoplasia in Wistar rats (Grasl-Kraupp et al., 

1994). 

Scopolamine Hydrobromide Trihydrate 

Scopolamine hydrobromide trihydrate dosing in mice 

did not result in significantly increased incidences of 

neoplasms at any site under any feeding protocol. 

The significantly lower incidences of liver neoplasms 

in dosed male and female mice fed ad libitum, which 

is interpretable as a chemical-related/protective effect 

against carcinogenesis, were not apparent when dosed 

mice were compared to the weight-matched controls. 

Overall Considerations 

Two of the four chemicals studied were convincingly 

found to cause neoplasms at three sites when evaluated under standard ad libitum feeding Conditions. 

The affected organs were the urinary bladder and 
D~e~ary ]~estriction,NTP T~ 460 95 

pancreas in male rats. and the liver in male mice. 

When dosed animals fed ad libitum were compared 

with weight-matched control animals, these three~ 

organs were again identified as sites of carcinogenicity. In contrast, none of these organs were sites of 

carcinogenesis after 2 or 3 years of dosing under the 

restricted feed protocols; an alternate site, the urinary 

bladder of female rats dosed for 32 months, was a 

target of carcinogenesis. 

The increased survival of the feed-restricted animals 

allowed more opportunities for dosing and additional 

time for neoplasm development; however, this did not 

result in an enhanced ability, to detect a carcinogenic 

response after 2 years.. No neoplasm response associated with chemical exposure under ad libitum 

feeding protocols was identified under restricted feed 

protocols after 2 to 3 years of chemical treatment. As 

anticipated, the weight-matched control comparisons 

provided the largest number of statistically significant 

increases in neoplasm incidences. This clearly 

indicates a method to increase the sensitivity of the 

bioassay to detect potential carcinogenic responses. 

The inclusion of concurrent weight-matched controls 

allows estimation of the effect of body weight changes 

on the "background" incidences of neoplasms in the 

dosed groups; three of the current studies demonstrate 

this. As discussed, the results for liver neoplasms in 

mice in the salicylazosulfapyridine study indicate that 

body weight has a greater influence on the neoplasm 

response than does exposure to the carcinogen. In 

this specific case, dosed mice fed ad libitum had an 

increased response; however, it is possible that the 

dose selected for a liver carcinogen could cause 

sufficient weight loss to mask the true carcinogenic 

potential of the chemical. Thus, a weight-matched 

control group can be employed to prevent false 

negatives for chemicals that suppress body weight. 

The statistically significant decreases in the incidence 

of liver neoplasm s in male and female mice in the 

scopolamine hydrobromide trihydrate study 

(Table 28), which might be interpreted as a chemicalrelated anticarcinogenic (protective) effect, were not 

apparent when the dosed mice were compared to the 

weight-matched controls. The previously discussed 

relationship between body weight and mouse liver 

neoplasms suggests this is an appropriate use of this 

control group. Because rodent carcinogenicity assays 

are used for identification of cancer prevention 

activity (Greenwald et al., 1990; Kelloff et al., 1993), 

recognition that chemical-related body weight 

decreases can cause decreased neoplasm incidences 

could preclude erroneous interpretations. 

Finally, ythe alleviation of age-related nephropathy in 

feed-restricted male rats in the salicylazosulfapyridine 

study prevented the development of treatment-associated neoplasms of the urinary tract that occurred in 

similarly dosed rats fed ad libitum. This observation 

allowed amore definitive conclusion regarding the 

cause of these responses in the ad libitum feeding 

protocol. 

The dose concentrations chosen for these comparisons 

were based upon the results of 13-week studies 

conducted under ad libitum feeding protocols. 

Because dietary restriction causes a variety of pleiotropic responses that affect the metabolism, distribution, and disposition of xenobiotics, it is probable that 

minimally toxic doses (maximum tolerated doses) 

under ad libitum feeding protocols will be altered 

under feed restriction conditions (Hart et al., 1,995). 

It remains to be seen what the results would be if the 

chemicals were evaluated at exposure concentrations 

that provide similar blood concentrations between 

ad libitum feeding and feed restriction regimens. It 

should be noted that in most cases, mean body 

weights of the dosed, feed-restricted animals were 

substantially less than those of the feed-restricted 

controls; this suggests that maximum tolerated doses 

were achieved in these restricted feed protocols. 

Whether a chemical evaluation is conducted under 

restricted feed or ad libitum feeding conditions, body 

weight changes should be expected in dosed animals, 

and consideration should be given to the inclusion of 

weight-matched controls to aid in the interpretation of 

these studies. 

CONCLUSIONS 

Butyl benzyl phthalate caused an increased incidence 

of pancreatic acinar cell neoplasms in ad libitum-fed 

male rats relative to ad libitum-fed and weightmatched controls. This change did not occur in rats 

in the restricted feed protocols after 2 years; however, 

acinar cell adenomas were observed in three exposed, 
96 Dietary Restriction, NTP TR 460 • 

feed-restricted males at 30 months. Feed restriction 

is known to influence the incidence of pancreatic 

acinar cell neoplasms and may have prevented the full 

expression of this chemical-induced effect. Butyl 

benzyl phthalate also caused an increased incidence of 

urinary bladder neoplasms in female rats in the 

32-month restricted feed protocol. The incidences of 

urinary bladder neoplasms were not significantly 

increased in female rats in any of the 2-year protocols, 

suggesting that the length of study, not body weight, 

was the primary factor in the detection of this carcinogenic response. 

Salicylazosulfapyridine caused an increased incidence 

of urinary bladder papillomas in male rats fed ad libitum relative to ad libitum-fed and weight-matched 

controls. This increase was associated with an 

increased incidence of urinary bladder calculi; the 

incidences of urinary bladder concretions, dilatation, 

and hyperplasia were also increased in dosed males, 

The incidences of urinary bladder papillomas and 

calculi were not increased in male rats receiving 

salicylazosulfapyridine and which were fed restricted 

diets. : 

In male mice, salicylazosulfapyridine caused an 

increased incidence of liver neoplasms relative to the 

ad libitum-fed and .weight-matched controls. This 

increased incidence did not occur in the restricted feed 

protocols. Liver neoplasms in mice are greatly 

influenced by body weight,~and the marked mean 

body weight reduction observed in dosed male mice in 

the restricted feed protocols may have overridden the 

carcinogenic response. 

, 

Neither t-butylhydroquinone nor scopolamine hydrobromide trihydrate caused increased neoplasm incidences under any ~of the experimenta! protocols. 

Regarding the future use of dietary restriction regimens in long-term studies; ordy limited conclusions 

can be drawn because 0nly four chemicals were. 

evaluated and none of these proved to be a strong 

carcinogen: However, the results of these studies are 

consistent with previous findings that dietary restric-., 

tion increases survival rates and decreases the incidences of neoplasms and nonneoplastic lesionsat a 

variety of sites in rats and mice. This association 

between reduced body weights and decreased neoplasm incidences underlines the necessity thai the 

doses selected for chronic studies not exceed 

"minimally toxic doses" so that no marked weight 

reductions (or increases) will occur in the dosed 

groups. Such body weight ~changes complicate 'the. 

detection of carcinogenic effects. 

A summary of the Technical Reports Review Subcommittee comments and the public discussion on this Technical Report appears on page 13. 
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8001-23-8, and 538-23-8) in Male F344/N Rats as 

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National Toxicology Program (NTP) (1994b). 

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NIH Publication No. 97-3374. U.S. Department of 

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Roebuck, B.D., Baumgartner, K.J., and 

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100 

Ross, M.H., and Bras, G. (1973). Influence of 

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. - - - - : - L " - - -

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~ _ _ _ _ 

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- _ _ 

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_ 

-~ 

_. _ . _ 

_ 

• 

-_ 

. . 

_ . 

.: - 2 

- 7-" Y 

-_ 

. _ 

. . . 

7_ ~ -S _- _ 

. 

Dietary Restriction, NTP TR 460 

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. 

.... - _..:~ .~ ~_~ 

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. -

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~ - . _ -

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J. Gerontol. 40,. 657-670~ -~ 

. 

-

.^ -. 

• . . . 

. • - .-~ -._ ..~ -

..-

- . 

.. 

-. 

. 5 
10~ ~ 

TABLE Ala 

TABLE ASh 

'TABLE A~a 

TABLE A2b 

TABLE A3a 

TABLE A3b 

AI~PENII~]IX A 

5~IAXY ©IF ]LES]I©N5 ]IN MALE I~.AT5 

~IN TIRE ~IETAXY XESTII~ICT~I©N 5T~..f~Y 

©~ BUTYIL ~ENZYIL I~I~IT~ALATE 

Summary of the Incidence of Neoplasms in Male Rats 

in the Dietary Restriction Study of Butyl ]Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 102 

Summary of the Incidence of Neoplasms in Male Rats 

in the Dietary Restriction Study of Butyl Benzyn Phthalate: 

2-Year and 30-Month Restricted Feed Protocols ............................ 107 

Stafistncan Analysis of Primary NeopRasms in Male Rats 

in the.Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols •. .................. 112 

Statistical Analysis of Primary Neoplasms in MaRe Rats 

in the Dietary Restriction Study of Butyl BenzyR Phthalate: 

2-'~'ear and 30-Month Restricted Feed Protocols ............................ 117 

Summary of the Incidence of Nonneoplastic Lesions in Mane Rats 

in the Dietary Restriction Study of Butyl Benzyl Phtha~ate: 

Ad Libitum Feeding and Weight-Matched ControRs Protocols ................... 122 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats 

in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

2-Year and 30-Month Restricted Feed Protocols ............................ 130 

!i I 
102 Dietary Restriction, NTP TR 460 

TABLE Ala 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad LibitumFed Control 

Weight-Matched 

Control 12,000 ppm 

Disposition Summary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Other 

60 

10 

19 

3 

28 

60 

10 

15 

1 

33 

1 

60 

10 

26 

2 

22 

Animals examined microscopically 

.. 

15-Month Interim Evaluation 

Endocrine System 

Adrenal medulla 

Pheochromocytoma benign 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

(10) 

1 (10%) 

(9) 

2 (22%) 

(10) 

(10) 

(10) 

1 (10%) 

(10) 

(10) 

(10) 

1 (10%) 

(10) 

1 (10%) 

Genital System 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

(10) 

3 (30%) 

4 (40%) 

(10) 

5 (50%) 

4 (40%) 

(10) 

5 (50%) 

4 (40%) 

Hematopoietic System . 

Spleen 

Histiocytic sarcoma 

(10) 

1 (10%) 

(10) (I0) 

Integumentary System 

Skin 

Keratoacanthoma 

(10) 

1 (10%) 

(10) (10) 

Systemic Lesions 

Multiple organs b 

Histiocytic sarcoma 

Leukemia mononuclear 

Mesothclioma malignant 

(10) 

1 (10%) 

(10) (10) 
Dietary Restriction, I~' TR ~6~ 103 

TABLE Ala 

Summary of the ~incidenee o~ Neo#~sms in MI~fle ~a~s in the Die~ry ]Restriction Stuc~y o~ ~utyi ~en~yi ~¢~aiate: 

Ad Libitum Feeding ~md Weigh*-Matched Controls ~'r'ot~eo~s (continued) .• '" 

Ad Libitum- Weight-Matched 

Fed Control Control 12,~ ppm 

25-Month ~nterira Evaluation (continued) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

Musculoskeletal System 

Nervous System 

• Respiratory System 

Special Senses System 

Urinary System 

2-Year Study 

Alimentary System 

Intestine large, colon (44) (49) (48) 

Carcinoid tumor malignant 1 (2%) 

Intestine large, rectum (49) (48) (50) 

Leiomyosarcoma 1 (2%) 

Intestine small, duodenum (50) (50) (50) 

Intestine small, jejunum (49) (50) (50) 

Leiomyoma 1 (2%) 

Intestine small, ileum (49) (50) (50) 

Liver (50) (50) (50) 

Carcinoma, metastatic, pancreas I (2%) 

Hepatocellular carcinoma 1 (2%) 

Hepatocellular adenoma 2 (4 %) 1 (2%) 4 (8%) 

Mesentery (7) (11 ) (5) 

Pancreas (50) (50) (50) 

Aeinus, adenoma 3 (6%) 10 (20%) 

Acinus, carcinoma 1 (2%) 1 (2%) 

Salivary glands (50) (50) (50) 

Schwannoma malignant l (2%) 

Stomach, forestomach (50) (50) (50) 

Stomach, glandular (50) (50) (50) 

Carcinoma 1 (2%) 

Cardiovascular System _ 

Heart (50) (50) (5o) 

Thymoma malignant, metastatic, thymus 1 (2%) 
104 I Dietary Restriction, NTP TR 460 

~-~ . 

TABLE Ala 

Summar.y.of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study. of Butyl Benzyl Phthalate: 

Ad Libitum-Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 12,000 ppm 

2-Year Study (continued) 

Endocrine System 

Adrenal cortex 

.-Carcinoma 

Adrenal medulla 

Pheochromocytoma malignant 

Pheochromocytoma benign 

Pheochromocytoma benign, nlultiple 

Islets, pancreatic 

Adenoma • , 

Pituitary gland 

Pars distalis, adenoma 

Pars distalis, adenoma, multiple 

Pars intermedia, adenoma 

Thyroid gland 

C-cell, adenoma 

Follicular cell, adenoma 

Follicular cell, carcinoma "~ 

General Body System 

None 

Genital System 

Preputial gland 

Adenoma 

Carcinoma 

Prffs/a~e 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

Hematopoietic System 

Bone marrow 

Carcinoma, metastatic, pancreas 

Lymph node . 

. ~Ilia_~, leiomyosarcoma, metastatic, 

intestine large, rectum 

Mediastinal, schwannoma malignant, 

metastatic, skin 

Lymph node,~mandibular 

Lymph node, mesenteric 

Spleen 

Thymus ~ - ~ 

Thymoma benign 

Thymoma malignant 

(50) .. 

(50) 

2 (4%) 

6 (12%) 

3 (6%) 

(50) 

5 (10%) 

(50) 

9 (18%) 

1 (2 %) 

(50) • 

5 (1o%) 

,, 

(50) 

" 4 (8%) 

1 (2%) 

(50) 

(50) 

(50) 

37 (74%) 

7-(14%) 

(5O) 

(23) 

1 (4%) 

(49) 

(50) 

(50) 

(49) 

(~o) 

1 (2%) 

(50) 

1 (2%) 

3 (6%) 

(50) 

I (2%) 

(50) 

9 (18%) 

1 (2%) 

I (2%) 

(50) 

3 (6%) 

(50) 

3 (6%) 

3 (6%) 

(50) 

(5O) 

(50) 

40 (80%) 

5 (10%) 

(50) 

1 (2%) 

(14) 

1 (7%) 

(48) 

(50) 

(50) 

(48) 

. . 

" 

, 

(50) 

(50) 

2 (4%) 

8 (16%) 

(5o) 

1 (2%) 

(49) 

10 (20%) 

(50) 

1 (2%) 

1 (2%) 

2 (4%) 

(50) 

(50) 

(50) 

(50) 

37 (74%) 

8 (16%) 

(50) 

(19) 

(50) 

(50) 

(50) 

(48) 

1 (2%) 

1 (2%) 

' . 
: : ~ ~ ~ ~-~;l~li~l~ ~. ~ . . . . . . . . ~ ...... , - - ~ - ; .................. ~ .... _ _ ...................... 

Dietary Restriction, NTP TR 460 

T~ Ala 

Summary of the ~nddence of Neop~mmS in IVJla~e Rats iin the Dietary Restriction Study of Butyl Benzy~ Phtha~ate: 

Ad Libitum ~eed~ng mad Weight-Matched Controls Protocds (cbntinued) 

Ad Libitum- Weight-Matche d 

Fed Control Control 12,~00 ppm 

2-Year Study (continued) 

Integumentary System 

Mammary gland 

Fibroadenoma 

Skin 

Basal cell adenoma 

Basal cell carcinoma 

Keratoacanthoma 

Squamous cell papilloma 

Subcutaneous tissue, fibroma 

Subcutaneous tissue, lipoma 

Subcutaneous tissue, melanoma malignant 

Subcutaneous tissue, schwannoma malignant 

Musculoskeletal System 

Bone 

Chordoma 

Osteoma 

Osteosarcoma 

Skeletal muscle 

Carcinoma, metastatic, pancreas 

Fibrosareoma 

Hemangiosarcoma 

Squamous cell carcinoma, metastatic, lung 

Nervous System 

Brain 

Oligodendroglioma malignant 

Spinal cord 

Respiratory System 

Lung 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, pancreas" 

Carcinoma, metastatic, adrenal cortex 

Chordoma, metastatic, bone 

Squamous cell carcinoma 

Thymoma malignant, metastatic, thymus 

(45) 

2 (4%) 

(50) 

1 (2%) 

2 (4%) 

1 (2%) 

5 (10%) 

1 (2 %) 

(50) 

l (2%) 

(I) 

1 (100%) 

(50) 

I (2%) 

(2) 

(50) 

1 (2%) 

(49) 

3 (6%) 

(50) 

2 (4%) 

1 (2%) 

! (2%) 

1 (2%) 

(50) 

I (2%) 

(4) 

1 (25%) 

1 (25%) 

l (25%) 

(50) 

2 (4%) 

(50) 

2 (4%) 

1 (2%) 

1 (2%) 

1 (2 %) 

(48) 

(50) 

2 (4%) 

I (2%) 

I (2%) 

I (2%) 

5 (I0%) 

I (2%) 

(2%) 

(50) 

I (2%) 

(50) 

(2) 

(5O) 

1 (2%) 

1 (2%) 
106 Dietary Restriction, NTP TR 460 

• 

TABLE Ala ~ 

Summary of the Incidence of Neoplas~ns in Male Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Spedal Senses System 

Zymbal's gland 

Carcinoma 

Urinary System 

Kidney 

Squamous cell carcinoma, metastatic, lung 

Renal tubule, adenoma 

Renal tubule, carcinoma 

Urinary bladd, er 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma maliguam 

Neoplasm Summary 

Total animals with primary neoplasms ¢ 

15-Month interim evaluation 

2-Year study 

Total primary neoplasms " 

15-Month interim evaluation 

2-Year study 

Total anirgals with benign neoplasms 

15-Month interim evaluation 

2-Ydar study 

Total benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with malignant neoplasms 

15-Month interim evaluation 

2-Year study 

Total malignant neoplasms 

15-Month interim evaluation 

-2-Year study 

" Total animals with metastatic neoplasms 

2-Year study 

Total metastatic neoplasms 

2-Year study 

Ad LibitumFed Control 

(50). 

1 (2%) 

(50) 

(50) 

31 (62%) 

1 (2%) 

8 

50 

12 

135 

8 

46 

11 

.94 

1 

37 

1 

41 

2 

2 

Weight-Matched 

Control 

(2) 

2 (100%) 

(50) 

1 (2%) 

i (2%) 

(50) 

(50) 

15 (30%) 

1 (2%) 

10 

50 

10 

108 

10 

47 

10 

76 

28 

32 

4 

7 

12,000 ppm 

(50) 

1 (2%) 

(50) 

(50) 

30 (60%) 

1 (2%) 

9 

50 

11 

136 

9 

47 

11 

91 

38 

45 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

c Primary neoplasms: ~1 neoplasms except metastatic neoplasms 
~ :'~ ~:--~=~-~:- ,_~ :~c~-- _~_~_.~-~_.~__~_~ . . . . . . . . . . . . . . . . - ....................... 

Die~ry ]~es~ricfion, NT~ TR ~0 

T~LE Alb 

S~ary o~ the Incidence o~ Neoplasms in ~a~e ~i~ats ~n the Dietary ~estr~ction S~udy . 

o~ Butyl Ben~y~ Phtha~ate: 2-Year and 30-Month ]~es~rlcted Feed ]Protocols a 

2-Year Restricted Feed 30-l~Ionth Restricted Feed 

0 ppm ~2,000 ppm 0 ppm 12,~ ppm 

Disposition Summary 

Animals initially in study 60 60 50 50 

15-Month interim evaluation 10 10 

Early deaths 

Accidental death 1 

Moribund 14 17 37 29 

Natural deaths 1 2 3 8 

Survivors 

Terminal sacrifice 34 31 10 13 

Animals examined microscopically 60 50 50 

15-Month Ynter~m Evaluation 

Genital System 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

(10) 

1 (10%) 

6 (60%) 

(1o) 

i (1o%) 

5 (~o~) 

Respiratory System 

Lung 

Alveolarforonchiolar adenoma 

(10) 

1 (10 % ) 

(10) 

Systemic Lesions 

Multiple organs b 

Leukemia mononuclear 

(lo) (lO) 

1 (lo%) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

Endocrine System 

General Body System 

Hematopoietic System ~ 

IntegumentarySystem 

Musculoskeletal System 

Nervous System 

Special Senses System 

Urinary System 
108 ' Dietary Restriction, NTPTR 460 

TABLE Alb 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study 

of.Butyl Benzyl Phthalate: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm ' 12,000 ppm 0 ppm 12,000 ppm 

2-Year and 30-Month Protocols 

Alimentary System 

Intestine large, colon (49) (48) (49) (48) 

Intestine large, cecum (49) (50) (49) (49) 

Intestine small, duodenum .(50) (50) (50) (49) 

Intestine small, jejunum (49) (50) (50) (48) 

.Carcinoma 2 (4%) " 

Intestine small, ileum (49) (50) (50) , '(49) 

Sarcoma ~ (2%) 

Liver (50) (50) (50) (49) 

Hepatocellular carcinoma 2 (4%) 

Hepatocellular adenoma (2%) 1 (2%) 

Hepatocellular adenoma, multiple 1 (2%) 

Histiocytic sarcoma 1 (2%) 

Mes~ntery (7) (9) (12) (9) 

Fibrous histi0cytoma ~ (11%) 

Sarcoma 1 (14%) ~ (1~%) Schwannoma malignant 

Oral mucosa (1) (2) O) 

~quamous cell carcinoma ~ 0OO%) I (50%) 

Squamous cell papilloma I (50%) 1 (100%) 

Pancreas (50) (50) (50) (49) 

Acinar cell, adenoma 1 (2%) 

Acinus, adenoma 1 (2%) 

1 (2%) 

Sarcoma, metastatic, mesentery 1 (2%) 

Salivary glands (50) (49) (50) (50) 

Fibrous histiocytoma, metastatic, skin ~ (2%) 

Schwannoma malignant I (2%) 

Acinus, adenorna, multiple 

(50) (50) Stomach, forestomach (5o) " (50) 

Leiomyosarcoma 1 (2%) 

Squamous cell papilloma 1 (2%) 1 (2%) • 4 (8%) 

Stomach, glandular (50) (50) (50) (49) 

Tongue (1) (i) (i) 

Squamous cell carcinoma ~ OO0%) 

Squamous cell papilloma 1 (100%) 

, : Cardiovascular System 

Heart (50) (50) (50) (50) 

Endocrine System 

Adrenal cortex (5O) (50) (50) (50) 

Adenoma I (2%) 

Adrenal medulla (50) • (50) (50) (50) 

Pheochromocytoma malignant 1 (2%) i (2%) 1 (2%) 

3 (6%) 4 (8%) 6 (12%) 6 (12%) Pheochromocytoma benign 

Pheochromocytoma benign, multiple 1 (2%) 

Bilateral, pheochromocytoma benign 2 (4%) 
Dietary ]Restriction, NT~ TR 460 109 

TABLE Alb 

Summary of the l~ncldence of Neoplasms ~u 1V~a~e RaN ~u the D~et,~'y ~estr~cfion Study 

of I~utyH ~enzyH Phtha~ate: 2-Year and 30-Month Restricted Feed ~Protoeo~s (continued) 

2-Year Restricted ~'eed 30-Month Restricted ~eed 

0 ppm 12,000 ppm 0 ppm ~2,~)00 ppm 

2- ~'ear and 30-Month Protocols (continued) 

Endocrine System (continued) 

Islets, pancreatic (50) (50) (50) (49) 

Adenoma 4 (8%) 1 (2%) 3 (6%) 5 (10%) 

Carcinoma 1 (2%) 1 (2%) 2 (4%) 2 (4%) 

Pituitary gland (46) (48) (50) (47) 

Pars distalis, adenoma 4 (9%) 7 (15%) 7 (14%) 4 (9%) 

Pars distalis, adenoma, multiple 1 (2%) 1 (2%) 

Thyroid gland (50) (50) (50) (50) 

C-cell, adenoma 4 (8%) 4 (8%) 4 (8%) 4 (8%) 

General Body System 

Peritoneum (2) (1) (2) (2) 

Genital System 

Epididymis (50) (50) (50) (50) 

Preputial gland (50) (50) (50) (50) 

Adenoma 1 (2%) 1 (2%) 2 (4%) 

Carcinoma 1 (2%) 

Prostate (49) (50) (50) (50) 

Adenoma 1 (2%) 

Carcinoma 1 (2%) 

Seminal vesicle (50) (50) (50) (48) 

Testes (50) (50) (50) (50) 

Bilateral, interstitial cell, adenoma 42 (84%) 42 (84%) 37 (74%) 38 (76%) 

Interstitial cell, adenoma 5 (10%) 6 (12%) 9 (18%) 7 (14%) 

Hematopoietic System 

Bone marrow (50) (50) .(50) (50) 

Hisfiocytic sarcoma 1 (2%) 

Lymph node (12) (19) (28) (23) 

Mediastinal, fibrous histiocytoma 1 (5%) 

Mediastinal, histiocytic sarcoma 1 (4%) 

Lymph node, mandibular (48) (47) (49) (49) 

Hisfiocyfic sarcoma 1 (2%) 

Lymph node, mesenteric (50) (50) (50) (49) 

Hisfiocytic sarcoma 1 (2%) 

Spleen (50) (50) (50) (49) 

Fibroma 1 (2%) 

Sarcoma 1 (2%) 

Thymus (46) (48) (46) (45) 

Thymoma benign 1 (2%) 1 (2%) 
110 Dietary Restriction, NTP TR 460 

TABLE Alb 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 .ppm 12,000 ppm 0 ppm 12,000 ppm 

2-Year and 30-Month Protocols (continued) 

Integumentary System 

Mammary gland (49) (50) (46) (49) 

Carcinoma 1 (2%) 

Fibroadenoma . 2 (4%) 3 (7%) 3 (6%) 

Skin (49) (50) (50) (48) 

Keramacanthoma 2 (4%) 2 (4%) 2 (4%) 2 (4%) 

Trichoepithclioma l (2%) 

Subcutaneous tissue, fibroma I (2%) 5 (10%) 2 (4%) 

Subcutaneous tissue, fibrosarcoma I (2%) 

Subcutaneous tissue, fibrous histiocytoma I (2%) 

Subcutaneous tissue, lipoma 1 (2%) 

Subcutaneous tissue, melanoma benign 2 (4%) 

Subcutaneous tissue, melanoma malignant I (2%) I.(2%) 

Musculoskeletal System 

Skeletal muscle (2) 

Fibrous histiocytoma i (50%) 

Fibrous histiocytoma, metastatic, skin. I (50%) 

Nervous System 

Brain (50) (50) (50) (49) 

Astrocytoma malignant I (2%) 2 (4%) 

Respiratory System 

Lung (50) (50) (50) (50) .. 

Alveolar foronchiolar adenoma 2 (4%) 

Alveolar/bronchiolar carcinoma i (2%) 1 (2%) 

• Sarcoma, metastatic, mesentery i (2%) 

Special Senses System 

Eye (1) (I) 

Schwarmoma malignant 1 000%) 

Zymbal's gland (t) (1) (I) 

Carcinoma i Ooo%) 1 (100%) I (100%) 

Urinary System 

Kidney (50) (5O) (50) (49) 

Urinary bladder (50) (5O) (50) (50) 

Transitional epithelium, carcinoma I (2%) 

Transitional epithelium, papilloma I (2%) I (2%) 
D~et~ry ]Restriction, NT~ TR 4~0 111 

TABLE A1b 

S~ary of the ~aci~eace of Neoplasms ~a ~ale Rats ~a the Dietary ~estr~ct~oa St~n~y 

of ~aty~ ]~enzy~ ~t~a~ate: 2-Year a~ 30-iV~ont~ Restricted Feed ~rotoco~s (continued) 

2-Year Restricted Feed 30-]~onth Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

2-Year and 30-Month Protocols (continued) 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Leukemia mononuelear 

Mesothelioma malignant • 

(50) 

21 (42%) 

2 (4%) 

(50) 

27 

1 

(54%) 

(2%) 

(50) 

1 (2%) 

39 (78%) 

2 (4%) 

(50) 

36 (72%) 

2 (4%) 

Neoplasm Summary 

Total animals with primary neoplasms ¢ 

15-Month interim evaluation 8 6 

2-Year and 30-month protocols 49 50 50 48 

Total primary neoplasms 

15-Month interim evaluation 8 7 

2-Year and 30-month protocols 100 112 144 134 , 

Total animals with benign neoplasms 

15-Month interim evaluation 8 6 

2-Year and 30-month protocols 48 50 47 46 

Total benign neoplasms 

~15-Month interim evaluation 8 6 

2-Year and 30-month protocols 69 75 88 85 

Total animals with malignant neoplasms 

15-Month interim evaluation 1 

2-Year and 30-month protocols 29 32 46 42 

Total malignant neoplasms 

15-Month interim evaluation 1 

2-Year and 30-month protocols 31 37 56 49 

Total animals with metastatic neoplasms 

2-Year protocol 1 1 

• Total metastatic neoplasms 

2-Year protocol 2 2 

a Number of animals examined microscopically ,at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

c Prima~y neoplasms: all neoplasms except metastatic neoplasms 
-- 

112 

{ 

TABLE A2a 

Statistical Analysis of Primary Neoplasms in Male Rats in th~ Dietary Restriction 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

Ad LibitumFed Control 

Adrenal Medulla: Benign Pheochromocytoma 

Overall rate a 9/50 (18%) 

Adjusted rate b 28.2% 

Terminal rate c 6/28 (21%) 

First incidence (days) 607 

Life table test d 

Logistic regression test d 

Fisher exact test d 

Adrenal Medulla: Benign or Malignant Pheochromocytoma 

Overall rate 

Adjusted rate : 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

~ 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rat~ 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Mammary Gland: Fibroadenoma ' 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

10/50 (20%) 

30.1% 

6/28 (21%) 

607 

2/50 (4 %) 

6.5% 

1/28 (4%) 

662 

2/50 (4%) 

6.5% 

1/28 (4%) 

662 

2/50 (4%) 

7.1% 

2/28 (7%) 

729 (T) 

12,000 ppm x 

Ad LibitumFed Control 

8/50 (16%) 

25.4% 

3/22 (14%) 

639 

P=0.589N 

P=0.475N 

P=0.500N 

10/50 (20%) 

33.3% 

5/22 (23%) 

639 

P=0.496 

P=0.573N 

P=0.598N 

-4/50 (8 %) 

14.2% 

2/22 (9%) 

464 

P=0,291 

P=0.339 

P=0.339 

4/50 (8%) 

14.2% 

2/22 (9%) 

464 

P=0.291 

P=0.339 

P=0,339 

0/50 (O%) 

0.0% 

0/22 (0%) _~ 

P=0.292N 

P=0.292N 

P=0,247N . 

"L~ ' " 

Dietary Restriction, NTP TR 460 

Study of Butyl Benzyl Phthalate: 

" ' 

12,000 ppm x 

Weight-Matched Weight-Matched 

Control Control 

3/50 (6%) 8/50 (16%) 

8.5% 25.4% 

2/33 (6%) 3/22 (14%) 

678 639 

P=0.049 " :,. 

P=0.086 

P=0,100 : 

3/50 (6%) 10/50 (20%) 

8.5% 33.3% 

2/33 (6%) 5/22 (23 %) -

678 639 

P=0.012 

P=0,028 

P=0,036 

1/50 (2%) 4•50 (8%) 

3.0% 14.2% 

1/33 (3%) 2/22 (9%) 

730 (T) 464 

P=0.107 

P=0.180 : 

P=0.181 

2/50 (4%) 4/50 (8%) 

6.1% 14.2% 

2/33 (6%) 2/22 (9%) 

730 (T) 464 

P=0.209 

P=0.331 

P=0.339 

3/50 (6%) 0/50 (0%) 

9.1% 0.0% 

3/33 (9%) 0/22 (0 %) 

730 (T) 

P=0.200N 

P=0.200N 

P=0.121N 

., 
-- 

:~:: ~- : ...... rli:rT~-,~ ......................................... 

Dietary Restriction, NTP TR ,~60 I13 ~ .~ 

TABLE ~a 

Statisfica9 Aua~ysis of Primary Neoplasms in IVIIa~e Rats in the DietarY Restriction Study of ~aty~ ~enzyl ~'htha~ate: 

Ad Libitum Feediug 

Pancreas: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pancreas: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

'First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

and Weight=IV~latched ControLs ~'rotoco~s (continued) ' 

12,000 ppm × 

Weight-Matched Weight-Matched 

Control Control 

0/50 (0%) 10/50 (20%) 

0.0% 41.0% 

0/33 (0%) 8/22 (36%) 

- 709 

P<0.001 

P<0.001 

P<0.001 

1/50 (2 %) 11/50 (22 % ) 

3.0% 42.7% 

1/33 (3%) 8/22 (36%) 

730 (T) 674 

P < 0.001 • 

P<0.001 

P=0.002 

1/50 (2%) 1/50 (2%) 

10/50 (20%) 10/49 (20%) 

24.8% 35.1% 

5/33 (15%) 5/22 (23%) 

479 684 

P=0.331 

P=0.565 

P=0.579 

3/50 (6 %) 0/50 (0%) 

8.5% 0.0% 

2/33 (6%) 0/22 (0%) 

678 

P=0.186N 

P=0.136N 

P=0.121N 

or Carcinoma 

Pancreatic Islets: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test' 

Fisher exact test 

Pituitary Gland (Pars Distalis): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Preputial Gland: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

~ 

Adenoma 

: 

~r . . . . . . . . .
 

Ad LibitumFed Control 

3/50 (6%) 

10.7% 

3/28 (11%) 

729 (T) 

3/50 (6 % ) 

10.7% 

3/28 (11%) 

729 (T) 

5/50 (1o%) 

17.9% 

5/28 (18%) 

729 (T) 

I0150 (20%) 

30.6% 

7/28 (25 %) 

438 

4/50 (8%) 

12.8% 

2/28 (7%) 

644 

. 

12,000 ppm × 

Ad LibitumFed Control 

10/50 (20%) 

41.0% 

8/22 (36%) 

709 

P=0.011 

P=0.016 

P=0.036 

11/50 (22 % ) 

42.7% 

8/22 (36%) 

674 

P=0.007 

P=0.014 

P=0.020 

1/50 (2%) 

4.5% 

1/22 (5%) 

729 (T) 

P=0.161N 

P=0.161N 

P=0.102N 

10/49 (2O%) 

35.1% 

5/22 (23 %) 

684 

P=0.431 

P=0.575N . 

P=0.579 

0/50 (O%) 

O.0% 

0/22 (0%) 

P=0.087N 

P=0.059N 

P=0.059N 
-- -- 

114 Dietary Restriction, NTP TR 460 

TABLE A2a 

Statistical Analysis of Primary Neoplasmsin Male Rats'in the DietaryRestriction Study of Butyl BenzylPhthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continuedi , , 

Ad LibitumFed Control 

12,000 ppm X 

Ad LibitumFed Control Weight-Matched 

Control 

12,000 ppm X 

Weight-Matched 

Control 

Preputial Gland: Adenoma or Carcinoma 

Overall rate 5/50 (10%) 0/50 (o%) 6/50 02%) 0/50 (0%) 

Adjusted rate 15.1% 0.0% 16.4% 0.0% 

Terminal rate 2/28 (7%) 0/22 (0%) 4/33 (12%) 0/22 (0%) 

First incidence (days) 614 607 

Life table test / P=0.045N P=0.042N

Logistic regression test P=0.032N P=0.020N 

Fisher exact test • P=O.O28N P--0.013N 

.Skin: Squamous Cell Papilloma or Keratoacanthoma 

Overall rate 3/50 (6%) 2/50 (4%) 1/50 (2%) 2/50 (4%) 

Adjusted rate 10.7 % 9.1% 2.8 % 9.1% 

Terminal rate 3/28 (I 1%) 2/22 (9%) 0/33 (0%) 2/22 (9%) 

First incidence (days) 729 (T) 729 (T) 719 730 (T) 

Life tabl~ test P=0.611N P-~0.351 

Logistic regression test P=0.61 IN P=0.406 

Fisher exact test P=0.500N P--0.500 

Skin: Basal Cell Adenoma or Carcinoma 

Overall rate 1/50 (2%) 3/50 (6%) 2/50 (4%) 3/50 (6%) 

Adjusted 2.5% 11.3% 6.1% 11.3% ' 

Terminal rate 0/28 (0%) 2/22 (9 %) 2/33 (6%) 2/22 (9%) 

First incidence (days) 593 639 730 (T) 639 

Life table test P=0.289 P=0.351 

Logistic regression test P=0.307 P--0.458 

Fisher exact test P=0.309 P=0.500 

Skin: Squamous Cell Papilloma, Keratoacanthomi~, Basal Cell Adenoma, or Basal Cell Carcinoma 

Overall rate .4/50 (8%) 5/50 (10%) 3/50 (6%) 5/50 (10%) 

Adjusted . 12.9 % 20.2 % 8.7 % 20.2% 

Terminal rate 3/28 (11%) 4/22 (18%) 2/33 (6%) 4/22 (18%) 

First incidence (days) 593 639 719 639 

Life table test P=0.403 P=O. 179 

Logistic regression test P=0.518 P=0.286 

Fisher exact test P=0.500 P=0.357 

Skin (Subcutaneous Tissue): Fibroma 

Overall rate 5/50 (10%) 5•50 (10%) 1/50 (2%) 5/50 (10%) 

Adjusted rate 16.0% 17.3% 3.0% 17.3% 

Terminal rate 3/28 (11%) 2/22 (9%) . 1/33 (3%) 2/22 (9%) 

First incidence (days) 614 668 730 (T) 668 

Life table test P=0.550 P=0.056 

Logistic regression test P=0.614N P--0.086 

Fisher exact test P=O.630N P=0.102 
115 

~ ",~ ........ ~ ~ .......... 

Dietary ~estrictioa, N'~ T ~ ~60 

T~LE A2a 

Statistical Analysis o~ Primary Neo#asms in Ma~e Rats in the Dietary ]Restriction Study of ~utyH Benzy~ Phthanate: 

Ad Libitum ~eeding and Weight-Matched Controls ~'otoco~s (continued) 

12,000 ppm x 12,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Testes: Adenoma 

Overall rate 44/50 (88%) 45•50 (90%) 45~50 (90%) 45/50 (90%) 

Adjusted rate 97.8% 100.0% 100.0% 100.0% 

Terminal rate 27/28 (96%) 22/22 (100%) 33/33 (100%) 22/22 (100%) 

First incidence (days) 477 464 422 464 

Life table test P=0.205 P=0.017 

Logistic regression test P=0.619 P=0.528 

Fisher exact test P=0.500 P=0.630N 

Thyroid Gland (C-cell): Adenoma 

Overall rate 5/50 (10%) 1/50 (2%) 3/50 (6%) 1/50 (2%) 

Adjusted rate 14.1% 3.6% 9.1% 3.6% 

Terminal rate 2/28 (7 %) 0/22 (0%) 3/33 (9%) 0/22 (0 %) 

First incidence (days) 438 709 730 (T) 7O9 

Life table test P=0.130N P=0.444N 

Logistic regression test P=0.105N P=0.387N 

Fisher exact test P=0.102N P=0.309N 

Thyroid Gland (Follicular Cell): Adenoma or Carcinoma 

Overall rate 0/50 (0%) 3/50 (6%) O/50 (0%) 3/50 (6%) 

Adjusted rate 0.0% 11.6% 0.0% 11.6% 

Terminal rate 0/28 (0%) 1/22 (5 %) 0/33 (0%) 1/22 (5%) 

First incidence (days) - 698 698 

Life table test P=0.096 P=0.070 

Logistic regression test P=0.121 P=0.099 

Fisher exact test P=0.121 P=0.121 

All Organs: Mononuclear Cell Leukemia 

Overall rate 31/50 (62%) 30/50 (60%) 15/50 (30%) 30/50 (60%) 

Adjusted rate - 71.8% 65.6% 34.9% 65.6% 

Terminal rate 16/28 (57%) 7/22 (32%) 6/33 (18%) 7/22 (32%) 

First incidence (days) 479 180 422 180 

Life table test P=0.478 P=0.002 

Logistic regression test P=0.492N P=0.003 

Fisher exact test P=0.500N P=0,002 

All Organs: Benign Neoplasms 

Overall rate 46•50 (92%) 47/50 (94%) 47/50 (94%) 47/50 (94%) 

Adjusted rate 100.0% 100.0% lOO.0% 100.0% 

Terminal rate 28/28 (100%) 22/22 (100%) 33/33 (100%) 22/22 (100%) 

First incidence (days) 438 464 422 464 

Life table test P=0.199 P=0.020 

Logistic regression test P=0.601 P=0.631 

Fisher exact test P=0.500 P=0.661N 
-- 

116 Dietary Restriction, NTP TR 460 

TABLE A2a 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

v 

12,000 ppm x 12,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

All Organs: Malignant Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Benign or Malignant Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

37/50 (74%) 

78.5 % 

18/28 (64%) 

255 

50/50 (100%) 

100.0% 

28/28 (100%) 

255 , 

38/50 (76%) 

77.5 % 

11/22 (50%) 

180 

P=0.342 

P=0.469 

P=0.500 

50/50 (100%) 

100.0% 

22/22 (100%) 

180 

P=0.260 

-f 

P= 1.000N 

28/50 (56%) 

59.2% 

14/33 (42%) 

339 

50/50 (100%) 

100.0% 

33/33 (100%) 

339 

38/50 (76%) 

77.5% 

11/22 (50%) 

180 

,P=O.Oli 

P=0.045 

P=0.028 ~,. 

50/50 (I00%) 

100.0% 

22/22 (100%) 

•180 

. P=0.027 

P = 1.000N 

(T)Terminal sacrifice. 

• ./ 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, liver, 

pancreas, pancreatic islets, pituitary gland, preputial gland, testes, and thyroid gland; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercu~rent mortality 

c Observed incidence at terminal kill . • 

d Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the ad libitum-fed or weight-matched controls and the 

exposed group. The life table test regards neoplasms in animals dying prior t o terminal kill a~ being (directly or ~indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall intidence rates. For all tests, a lower incidence in an 

exposure group is indicated by N. 

• Not applicable; no neoplasms in animal group 

f Value of statistic cannot be computed. . . e 
117 

77 " ~ ' ~ = ~ ~}~ ~ ? ' ~ ......... ~ ' ~ ............ 

Die~ry ]Restriction, NIT TR 4~ 

TABLE A2b 

Stat~stic~a~ Analysis of Primary Neoplasms in Male Rats ~n the Dietary Restriction Study of BUtyl Benzy~ Phtha~ate: 

2-Year and 30-Month Restricted Feed Protocols 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

Adrenal Medulla: Benign Pheochromocytoma 

Overall rate a 3/50 (6%) 4/50 (8%) 9/50 (18%) 6/50 (12%) 

Adjusted rateb 7.3% 14.0% 47.6% 38.5% 

Terminal rate e 1/34 (3%) 4/31 (13%) 3/10 (30%) 4/13 (31%) 

First incidence (days) 467 729 (T) 730 807 

Life table test d P =0.458 P =0.165N 

Logistic regression test d P=0.502 P=0.239N 

Fisher exact test d P=0.500 P=0.288N 

Adrenal Medulla: Benign or Malignant Pheochromocytoma 

Overall rate 4/50 (8 %) 5/50(10%) 9150(18%) 6150 (12%) 

Adjusted rate 9.9 % 17.1% 47.6% 38.5% 

Terminal rate 1/34 (3 %) 5/31 (16%) 3/10 (30%) 4/13 (31%) 

First incidence (days) 467 729 (T) 730 807 

Life table test P=0.447 P=0.165N 

Logistic regression test P=0.500 P=0.239N 

Fisher exact test P=0.500 P=0.288N 

Liver: Hepatoeellular Adenoma or Carcinoma 

Overall rate 1/50 (2 %) I150 (2%) 3150(6%) 0/49 (0%) 

Adjusted rate 15.0% 0.0% 

Terminal rate 1/10 (10%) 0/13 (0%) 

. ~ First incidence (days) 710 

Life table test P = 0.117N 

Logistic regression test P=0.128N 

Fisher exact test P=0.125N 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 0/50 (0%) 0/50 (0%) I150 (2%) 3/50 (6%) 

Adjusted rate 10.0% 17.4% 

Terminal rate 1/10 (10%) 1/13 (8%) 

First incidence (days) 897 (T) 827 

Life table test P=0.428 

Logistic regression test P=0.350 

Fisher exact test P=0.309 

Mammary Gland: Fibroadenoma 

Overall rate 2/50 (4%) 0/50 (0%) 3/50 (6%) 3/50 (6%) 

Adjusted rate 14.6% 13.4% 

Terminal rate 0110 (0%) 1/13 (8%) 

First incidence (days) 768 687 

Life table test P=0.619N 

Logistic regression test P=0.649 

Fisher exact test P=0.661N 
-- 

118 

TABLE A2b 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary 

2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

0 ppm 12,000 ppm 

Man~iary Gland: Fibroadenoma or Carcinoma 

Overall rate 2/50 (4%) 0/50 (0%) 

Adjusted rate 5.7% 0.0% 

Terminal rate 2/34 (6%) 0/31 (0%) 

First incidence (days) ' 729 (T) -

Life table test 

Logistic regression test 

Fisher exact test • 

Pancreas: Adenoma 

Overall rate 0/50 (0%) 0/50 (0%) 

Adjusted ~te' 0.0% 0.0% 

Terminal 'rate 0/34 (0%) 0/31 (0%) 

First incidence (days) _ - -

Life table test 

Logistic regression test 

Fisher exact test 

Pancreatic Islets: Adenoma 

Overall rate 4/50 (8%) 1/50 (2%) 

Adjusted rate 11.0% 4.8% 

Terminal rate 2/34 (6%) 1/31 (3%) 

First incidence (days) • 719 730 (T) 

Life table test P=0.217N 

Logistic regression test P=0.206N 

Fisher exact test P=0.181N 

Pancreatic Islets: Adenoma or Carcinoma 

Overall rate 5/50 (10%) 2/50 (4%) 

Adjusted rate 13.8% 9.5% 

Terminal rate 3/34 (9%) 2/31 (6%) 

First incidence (days) 719 730 (T) 

Life table test P=0.263N 

Logistic regression test P=0.255N 

Fisher exact test P=0.218N 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 4/46 (9%) 8/48 (17%) 

Adjusted rate 11.0% 25.5% 

Terminal rate • 3/33 (9%) 5/31 (16%) 

First incidence (days) 681 350 

Life table test P=0.158 

Logisti c regressio n test P=0.227 

Fisher exact test : P=0.199 

Restriction 

Dietary Restriction, NTP TR 460 

Study of Butyl Benzyl Phthalate: 

~ 

30-Month Restricted Feed 

0 ppm 

3/50 (6%) 

I4.6% 

0/10 (0%) 

768 

0/50 (0%) 

0.0% 

0/I0 (0%) 

3/50 (6%) 

15.1% 

0/10 (0%) 

755 

5/50 (10%) 

28.1% 

I/I0 (10%) 

755 

7/50 (14%) 

32..9% 

2/10 (20%) 

597 

12,000 ppm 

4/50 (8%) 

18.0% 

1/13 (8%) 

687 

P=0;568 

P=0.483 

P=0.500 

3/49 (6%) 

23.1% 

3/13 (23%) 

897 (T) 

P=0.163 

P=0.163 

P=0.117 

5/49 (10%) 

32.9% 

4/13 (31%) 

723 

P=0.465 

P=0.371 

P=0.346 

7/49 (14%) 

38.4% 

4/13 (31%) 

723 

P=0.518 

P=0.385 

P=0.365. 

5/47 ( 11% ) 

35.4% 

4/13 (31%) 

869 

P=0.281N 

P=0.407N 

P=0.424N 
-- 

Dietary Restriction, NTP TR ,060 ~9 

TABLE A2b 

S~t~stic~d An~flys~s of ]?rimary Neoplmms in Male ~a~ ~n the Dietary ges~riction Study of It~uty~ ~enazy~ ~Phth~flate: 

2-Yesw and 3D-Month R e s t r i c t e d F e e d ]l~rotoco~s (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

Skin: Keratoacanthoma or Trichoepithelioma 

Overall rate 2/50 (4%) 2/50 (4%) 3/50 (6%) 

Adjusted rate 23.3% 

Terminal rate 2/10 (20%) 

First incidence (days) 768 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): Fibroma 

Overall rate 0/50 (0%) 1/50 (2%) 5/50 (10%) 

Adjusted rate 26.4% 

Terminal rate 0/10 (0%) 

First incidence (days) 703 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): Fibroma, Fibrosarcoma, or Fibrous Histiocytoma 

Overall rate 0/50 (0%) 2/50 (4%) 6•50 (12%) 

Adjusted rate 31.3% 

Terminal rate 0/10 (0%) 

First incidence (days) 703 

Life table test 

Logistic regression test 

Fisher exact test 

Stomach (Forestomach): Squamous Cell Papilloma 

Overall rate 1/50 (2%) 1/50 (2%) 4/50 (8%) 

Adjusted rate 19.1% 

Terminal rate 1/10 (10%) 

First incidence (days) 719 

Life table test \ 

Logistic regression test 

Fisher exact test 

Testes: Adenoma 

Overall rate 47/50 (94%) 48/50 (96%) 46/50 (92%) 

Adjusted rate 100.0% 100.0% 100.0% 

Terminal rate 34/34 (100%) 31/31 (100%) 10/10 (100%) 

First incidence (days) 549 530 516 

Life table test P=0.252 

Logistic regression test P=0.775N 

Fisher exact test P=0.500 

2/50 (4%) 

5.2% 

0/13 (0%) 

521 

P=0.453N 

P=0.509N 

P=0.500N 

2/50 (4%) 

8.2% 

0/13 (0%) 

674 

P=0.1g0N 

P=0.228N 

P=0.218N 

2/50 (4%) 

8.2% 

0/13 (0%) 

674 

P=0.105N 

P=0.142N 

P=0.134N 

O/50 (0%) 

0.0% 

0/13 (0%) 

P=0.060N 

P = 0.067N 

P=0.059N 

45/50 (90%) 

100.0% 

13/13 (100%) 

498 

P=O.302N 

P=0.390 

p=0.'500N 
120 Dietary Restriction, NTP TR 460 

TABLE A2b 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

2-Year and 30-Month Restricted Feed Protocols (continued) , ~ . . 

2-Year Restricted Feed 

Thyroid Gland (c-C~II): Adenoma 

Overall rate 

Adjusted rate. 

Terminal rate 

First incidence (day~) 

Life table .test 

Logistic regression test 

Fisher exact test 

All Organs: M0nonuclear Cell Leukemia 

Overall rate 

Adjusted rate 

Terminal rate" 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Benign Neoplasms 

Overall rate 

,, Adjusted rate 

Terminal rate. 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test . 

• -- ._.~. 

All Organs: Malignant Neoplasms 

Overall rate 

Adjusted rate -

Terminal rate - ~_ 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

30-Month Restricted Feed 

0 ppm 12,000 ppm 

4/50 (8%) " " ~1)50 (8%) 

14.3% 16.6% 

0/10 (0%) 1/13 (8%) 

701 498 

P=0.605N 

P=0.639N 

P=0.643N 

. . . . 

39/50 (78 %) 36/50 (72%) 

91.!% 88.8% 

7/10 (70%) 9/13 (69%) 

569 . 498 

P=0.276N 

P=0.375N 

P=0.322N 

47/50 (94%) . 46/50 (92 %) 

100,0% 100.0% 

10/10 (100%) 13/13 (100%) 

516 498 

P=0.301N 

P=0.224 

P=0.500N 

46/50 (92 %) 42/50 (84%) 

95.8% 93.0% 

8/10 (80%) 10/13 (77%) 

516 498 

P=0.228N 

P=0.585N 

P=0.178N 

0 ppm 

4/50 (8%) 

9.9% 

0/34 (0%) 

549 . 

21/50 (42%) 

47.4% 

12/34 (35 %) 

583 

. 

48/50 (96%) 

100.0% 

34/34 (100%) 

467 

29/50 (58%) 

59. 1% 

15/34 (44%) 

432 

12,000 ppm 

4/50 (8%) 

16.2% 

3/31 (10%) 

619 

P=0.598 

P=0.643N 

P=0.643N 

27/50 (54%) 

60.7% 

12/31 (39%) 

530 

P=0.145 

P=0.343 

P=0.158 

50/50 (100%) 

100.0% 

31/31 (100%) 

350 

P=0.208 

P=0.162 

P=0.247 

32/50 (64 %) 

68.9% 

15/31 (48%) 

530 

P=0.281 

P=0.418 

P=0.341 
:_.__ :7 ~.~-~ ~ - - ~ - ~ - ~ .......... ,;,: ........................ , :~ . . . . . . . . . . . . . ~ 

Dietary Restriction, NTP TR 460 ~121 

TABLE A2b 

•Statistican Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of Butyl ~enzyn Phthalate: 

2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

All Organs: Benign or Malignant Neoplasms 

Overall rate 49/50 (98%) 50/50 (100%) 50/50 (100%) 48/50 (96%) 

Adjusted rate 100.0% 100.0% 100.0% 100.0% 

Temainal rate 34/34 (100%) 31/31 (100%) 10/10 (100%) 13/13 (100%) 

First incidence (days) 432 350 516 498 

Life table test P=0.264 P=0.280N 

Logistic regression test P=0.456 -f 

Fisher exact test P=0.500 P=0.247N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, liver, 

lung, pancreas, pancreatic islets, pituitary gland, testes, and thyroid gland; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrem mortality 

¢ Observed incidence at terminal kill 

d Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the controls and that exposed group. The life table 

test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in an exposure group is indicated by 

N. 

e Not applicable; no neoplasms in animal group 

f Value of statistic cannot be computed. 

I 
122 Dietar~ Restriction, NTP TR 460 

TAnI~ A3a 

Summary of the Incidence of Nonneoplastic Lesiom in Male Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Control 12,000 ppm 

Disposition Summary 

Animals iaitially in study 60 60 60 

15-Month interim evaluation 10 10 10 

Early deaths 

' Moribund 19 15 26 

Natural deaths 3 1 2 

Survivors 

Terminal sacrifice 28 33 22 

Other 1 

Animals examined microscopically 60 60 60 

15-Month Interim Evaluation 

Alimentary System 

Intestine large, colon (I0) (I0) (I0) 

Parasite metazoan • 1 (I0%) I (10%) 

Intestine large, rectum (I0) (9) (~) 

Parasite metazoan 2 (20%) 2 (22%) 1 01%) 

Intestine large, cecum (I0) (I0) (10) 

Edema I (10%) 

Liver (10) (10) (10) 

Angiectasis 1 (10%) 

Easophilic focus : 6 (60%) 2 (20%) 1 (10%) 

Clear cell focus 2 (20%) 1 (10%) I (10%) 

Eosinophilic focus 3 (30%) 2 (20%) 

Granuloma I (10%) 

Hemorrhage 1 (10%) 

Hcpatodiaphragmatic nodule 3 (30%) I (10%) 

Inflammation, subacute 1 00%) 1 00%) 

Mixed cell focus I (10%) 

• 

Bile duct, hypcrplasia 5 (50%) 5 (50%) 

Hcpatocyte, vacuolization cytoplasmic 2 (20%) 3 (30%) 

Kupffcr ceil, pigmentation i (1o%) 

Lobulcs, necrosis 2 (20%) 

Mesentery (4) (4) (,I) 

Accessory spleen l (25%) 1 (100%) 

Fibrosis I (25%) 

Fat, necrosis 4 (100%) 3 (75%) 

Pancreas (10) (10) (1o) 

Atrophy 3 (30%) , 6 (60%) 4 (4o%) 

Cytoplasmic alteration 1 (1o%) 

Stomach, glandular (IO) (1o) (1o) 

Mucosa, hyperplasia 1 (10%) 

Cardiovascular System 

Heart (1o) (1o) (10) 

Cardiomyopathy 8 (80%) 5 (5o%) 6 (6O%) 

a Number of animals examined microscopically at the site and the number of animals with lesion ¢ 
~ ~ , . . . . . . : - ~ . ~ : ; ~ ............. .~ ~ 

Dietary ~estr~cfion, NTP TR 460 a23 

TAI~L~ A3a 

Summary of the Incidence of Nonneo#mfic Lesions ~n MaLe ~a~s ~n the D~et,m'y ~estr~ction Study 

of ~uty~ ~enzy~ Phtha~ate: Ad Libitum ~eed~ng and Weight-Matched Controls ]Protocols (continued) 

Ad LibitutnFed Control 

15-Month ~nterim Evaluation (continued) 

Endocrine System 

Adrenal cortex (10) 

Accessory adrenal cortical nodule 4 (40%) 

Degeneration, fatty 

Hyperplasia, focal 1 (10%) 

Hypertrophy, focal 

Pituitary gland (9) 

Pars distalis, angiectasis 

Pars distalis, cyst 1 (11%) 

Pars distalis, hemorrhage 

Pars distalis, hyperplasia, focal 3 (33%) 

Thyroid gland (10) 

Uldmobranchial cyst 

C-cell, hyperplasia 

Follicle, cyst 

Weight-Matched 

Control 

(10) 

, 3 (30%) 

1 (10%) 

(10) 

1 (10%) 

1 (10%) 

3 (3O%) 

(10) 

1 (10%) 

12,000 ppm 

(10) 

3 (30%) 

1 (10%) 

(10) 

1 (10%) 

1 (10%) 

2 (2O%) 

(10) 

2 (2O%) 

1 (10%) 

Genital System 

Epididymis (10) (10) (10) 

Atypia cellular 1 (10%) 3 (30%) 2 (20%) 

Hypospermia 1 (10%) 

Prepudal gland (10) (10) (10) 

Inflammation, chronic 6 (60%) 7 (70%) 6 (60%) 

Inflammation, suppurative 2 (20%) 1 (10%) 

Prostate (10) (10) (10) 

Corpora amylacea 1 (10%) 2 (20%) 7 (70%) 

Inflammation, suppurative 5 (50%) 7 (70%) 4 (40%) 

Epithelium, hyperplasia 1 (10%) 

Testes (10) (10) (10) 

Interstitial cell, hyperplasia 5 (50%) 4 (40%) 4 (40%) 

Hematopoietic System 

Lymph node (2) (1) 

Deep cervical, hemorrhage 1 (50%) 

Deep cervical, pigmentation 1 (50%) 

Mediasfinal, hemorrhage 2 (100%) 1 (100%) 

Mediastinal, pigmentation 2 (100%) 1 (100%) 

Lymph node, mandibular (10) (10) (10) 

Ectasia 1 (10%) 

Hemorrhage 2 (20%) 2 (20%) 

Hyperplasia, lymphoid I (10%) 1 (10%) 

Lymph node, mesenteric (10) (10) (10) 

Ectasia 1 (10%) 

Hemorrhage 

Hyperplasia, lymphoid 
•124 • " Dietary Restriction, NTP TR 460 

TABLE A3a , 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

. • Ad LibitumFed Control Weight-Matched 

Control • 12,000 ppm 

15-Month Interim Evaluation (col~tinued) 

Hematopoietic System (continued) 

Spleen . (10) 

Hematopoietic cell proliferation 2 (20%) 

Pigmentation, hemosiderin 10 (100%) 

Thymus (10) 

Cyst 1 (10%) 

(10) 

9 (90%) 

(10) 

(10) 

~ (10%) 

7 (70%) 

(9) 

,~., 

Musculoskeletal System 

Bone 

Femur, osteopetrosis 

(10) . 

1 (10%) 

(10) (10) 

Respiratory System 

Lung , 

Hemorrhage 

Infiltration cellular, histiocyte 

Alveolar epithelium, hyperplasia 

Nose 

Exudate 

Foreign body 

Fungus 

Mucosa, hyperplasia 

Mucosa, metaplasia, squamous 

(lO) 

! (lO%) 

2 (20%) 

1 (lO%) 

(lO) 

(10) 

1 (10%) 

(10) 

3 (30%) 

1 (10%) 

2 (20%) 

1 (10%) 

2 (20%) 

(10) 

2 (20%) 

1 (10%) 

(10) 

Special Senses System 

Eye 

Cataract 

Hemorrhage 

Retina, degeneration 

(1) 

1 (lOO%) 

1 (1oo%) 

1 (loo%) 

Urinary System 

Kidney 

Nephropathy. 

Renal tubule, pigmentation 

(io) 

lO (lOO%) 

lO (lOO%) 

(10) 

9 (90%) 

10 (100%) 

(1o) 

9 (9o%) 

lO (1oo%) 

°Systems Examined With No Lesions Observed 

General Body System 

Integumentary System 

Nervous System 
125 

5: -.-~.=---z::.~r --~ 2-_ :--- . ....................... ,, ~ ......... ~'~-~'~ 7:'~ , 

Dietary Restriction, NT]? TR 460 

~&BLE A3a 

Sunm~ary o~ the Incidence of Nonneopl~stic Lesioras in Mane l~ats in the Dietary ~estrictlon Study 

of butyl ~enzyl ]Phthalate: Ad Libitum ~eeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 12,000 ppm 

2- ~rear Study 

Alimentary System 

Intestine large, colon (44) (49) (48) 

Dilatation 1 (2%) . 

Parasite metazoan 5 (11%) 5 (10%) 4 (8%) 

Intestine large, rectum (49) (48) (50) 

Parasite metazoan 10 (20%) 6 (13%) 8 (16%) 

Intestine large, cecum (47) (50) (50) 

Edema 3 (6%) 

Parasite metazoan 1 (2%) 

Ulcer 1 (2%) I (2%) 

Intestine small, ileum (49) (50) (50) 

Parasite metazoan 1 (2%) 

Liver (50) (50) (50) 

Angiectasis 4 (8%) 2 (4%) 

Basophilie focus 22 (44%) 20 (4O%) 14 (28%) 

Clear cell focus 15 (30%) 8 (16%) 8 (16%) 

Cyst 2 (4%) 

Degeneration, cystic 7 (14%) 7 (14%) 6 (12%) 

Developmental malformation 1 (2%) 

Eosinophilic focus 3 (6%) 10 (20%) 4 (8%) 

Granuloma 7 (14%) 

Hematopoietic cell proliferation 1 (2%) 

Hemorrhage 1 (2%) 

Hepatodiaphragmatic nodule 1 (2%) 4 (8%) 7 (14%) 

Inflammation, subacute 4 (8%) 

Mixed cell focus 6 (12%) 6 (12%) 7 (14%) 

Bile duct, hyperplasia 39 (78%) 34 (68%) 27 (54%) 

Centrilobular, necrosis 1 (2%) 2 (4%) 

Hepatocyte, vacuolization cytoplasmic 6 (12%) 9 (18%) 4 (8%) 

Kupffer cell, pigmentation 1 (2%) 6 (12%) 

Lobules, necrosis 2 (4%) 2 (4%) 1 (2%) 

Mesentery (7) (11) (5) 

Accessory spleen 1 (9%) I (20%) 

Angiectasis 1 (14%) 

Fat, necrosis 5 (71%) 8 (73%) 3 (6O%) 

Pancreas (50) (50) (50) 

Atrophy 29 (58%) 31 (62%) 29 (58%) 

Acinus, cytoplasmic alteration 1 (2%) 1 (2%) 

Acinus, hyperplasia, focal 4 (8%) 2 (4%) 12 (24%) 

Stomach, forestomach (50) (50) (50) 

Edema 1 (2%) 3 (6%) 

Ulcer 2 (4%) 1 (2%) 3 (6%) 

Mucosa, hyperplasia 2 (4%) 1 (2%) 1 (2%) 

Stomach, glandular 

Edema 

(50) 

2 (4%) 

(50) 

1 (2%) 

(50) 

Erosion 

Ulcer 

Tongue 

Epithelium, hyperplasia 

2 

2 

(4%) 

(4%) I (2%) 

1 

1 

(1) 

1 

(2%) 

(2%) 

(100%) 
126 Dietary Restriction, NTP TR 460 

TABLE A3a 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study " 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 12,000 ppm 

2-Year Study (continued) 

Cardiovascula~ System 

Blood vessel (50) (50) (50 ) 

Hypertrophy 2 (4%) 2 (4%) 

Inflammation, subacute 2 (4%) 2 (4%) 

Heart (50) (50) (50) 

Cardiomyopathy 32 (64%) 24 (48%) 34 (68%) 

Inflammation, chronic 1 (2%) " 

Thrombosis 2 (4%) 5 (10%) 

Schwann cell, hyperplasia 1 (2%) 

Endocrine System 

Adrenal cortex (50) (50) (50) 

Accessory adrenal cortical nodule 

Angiectasis 

Degeneration, fatty 

Hemorrhage 

Hyperplasia, focal 

Hypertrophy, focal 

11 (22%) 

3 (6%) 

1 (2%) 

7 (14%) 

2 (4%) 

I0 

8 

3 

I 

9 

3 

(20%) 

(16%) 

(6%) 

(2%) 

(18%) 

(6%) 

6 (12%) 

2 (4%) 

8 (16%) 

1 (2%) 

4 (8%) 

1 (2%) 

. Vacuolization cytoplasmic 

Adrenal medulla 

Hyperplasia 

Islets, pancreatic 

! (2%). 

(50) 

6 (12%) 

(50) 

(50) 

6 

(50) 

(12%) 

(50) 

12 (24%) 

(50) 

Hyperplasia 

Parathyroid gland 

3 (6%) •

(49) (48) 

2 (4%) 

(49) 

Hyperplasia 

Pituitary gland 

Pars distalis, angiectasis 

Pars distalis, cyst 

Pars distalis, hyperplasia 

Pars distalis, hyperplasia, focal 

Pars distalis, necrosis 

(5O) 

3 (6%) 

3 (6%) 

I (2%) 

7 (14%) 

(50) 

3 

3 

(6%) 

(6%) 

(18%) 

i (2%) 

(49) 

2 (4%) 

2 (4%) 

5 (1o%) 

1 (2%) 

Pars intermedia, angiectasis 3 (6%) 2 (4%) 

Pars intermedia, cyst 1 (2%) I (2%) 

Pars intermedia, hyperplasia 

Thyroid gland (50) 

2 (4%) 

(50) (50) 

Hyperplasia 

Ultimobranchial cyst 

C-cell, hyperplasia 

Follicle, cyst 

Follicular cell, hyperplasia 

2 (4%) 

4 (8%) 

2 (4%) ~.. 

I (2%) 

1 (2%) 

12 (24%) 

1 (2%) 

2 (4%) 

7 (14%) 

4 (8%) 

General Body System 

None 
127 

~ ' ~ . ~ r ~ .............................. ~ ................. 

Die~ry ~est~cfioa, ~ T~ ~ 

T~LE A3a 

S~ary o~ the ]Mci~ence o~ Nonaeo#astic lLesiom ~a Ma~e ~ats ~ t]~e Dietary ~estr~ct~oa Stagy 

o~ ~ty~ ~en~yH P~t~a~ate: Ad L/b/turn Yee~ag aa~ We~g~t-iV~atc~ed Co~tro~s ~r'otoco~s (continued) 

Ad Libitum- Weight-Matched 

~'ed Control Control 12,000 ppm 

i 

2- Year Study (continued) 

Genital System 

Epididymis (50) (50) (50) 

Atypia cellular 28 (56%) 30 (60%) 37 (74%) 

I Hypospermia 34 (68%) 40 (80%) 43 (86%) 

Preputial gland (50) (50) (50) 

Ectasia 11 (22%) 9 (18%) 14 (28%) 

' Hyperplasia 1 (2%) 2 (4%) 1 (2%) 

= Inflammation, chronic 25 (50%) 21 (42%) 17 (34%) 

Inflammation, suppurative 9 (18%) 5 (10%) 6 (12%) 

Prostate (50) (50) (50) 

Corpora amylaeea 17 (34%) 24 (48%) 30 (60%) 

Fibrosis 1 (2%) 

Inflammation, suppurative 29 (58%) 27 (54%) 19 (38%) 

Epithelium, hyperplasia 4 (8%) 5 (10%) 3 (6%) 

I Seminal vesicle (50) (50) (50) 

~ Dilatation 4 (8%) 1 (2%) 

Testes (50) (50) (50) 

Interstitial cell, hyperplasia 4 (8%) 5 (10%) 6 (12%) 

Seminiferous tubule, atrophy 2 (4%) 2 (4%) 4 (8%) 

Hematopoietic System 

Bone marrow (50) (50) (50) 

Hypereellularity I (2%) 2 (4%) 

Myelofibrosis 2 (4%) 3 (6%) 5 (10%) 

Lymph node (23) (14) (19) 

Iliac, hemorrhage 1 (4%) 

Iliac, hyperplasia, lymphoid 1 (4%) 

Iliac, pigmentation I (4%) 

Mediastinal, hemorrhage 5 (22%) 4 (29%) 

Mediastinal, hyperplasia, lymphoid 

Mediastinal, pigmentation (22%) 

1 (7%) 

4 (29%) 5 (26%) 

Pancreatic, hemorrhage 1 (5%) 

Pancreatic, hyperplasia, lymphoid 1 (7%) 

Pancreatic, pigmentation 1 (7%) 2 (11%) 

Renal, hyperplasia, lymphoid 1 (4%) 

Renal, pigmentation 2 (9%) 2 (11%) 

Lymph node, mandibular (49) (48) (50) 

Congestion 1 (2%) 

Ectasia 6 (12%) 4 (8%) 4 (8%) 

Hemorrhage 4 (8%) 8 (17%) 6 (12%) 

Hyperplasia, lymphoid 3 (6%) 5 (10%) 5 (10%) 

Pigmentation 2 (4%) 7 (15%) 

Lymph node, mesenteric (50) (50) (50) 

Ectasia 6 (12%) 1 (2%) 1 (2%) 

Hemorrhage 1 (2%) 3 (6%) 4 (8%) 

Hyperplasia, lymphoid 2 (4%) 
128 Dietary Restriction, NTP TR 460 

TAnLE A3a 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the. Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and. Weight-Matched Controls Protocols (cgntinued) " 

Ad Libitum- Weight-Matched 

Fed Control Control 12,000 ppm 

2-Year Study (continued) 

Itematopoietie System (continued) 

Spleen (50) • (50) (5o) 

Fibrosis 5 (10~o) 7 (14%) 10 (20%) 

Granuloma 1 (2%) . 

Hematopoietic cell proliferation 2 (4%) 6 (12%) 7 (14%) 

Metaplasia, osseous 1 (2%) 

Necrosis 1 (2%) 1 (2%) 

Pigmentation, hemosiderin 14 (28%) 2 (4%) 6 (12%) 

Thymus (49) (48) (48) 

Hemorrhage 1 (2%) 

Integumentary System 

Mammary gland 

Hyperplasia 

Skin 

Acanthosis 

(45) 

10 (22%) 

(50) 

(49) 

8 (16%) 

i5o) 

1 (2%) 

(48) 

12 (25%) 

(50) 

10 (20%) 

Cyst epithelial inclusion 

Hemorrhage 

Hyperkeratosis 

Inflammation,- chronic 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

13 (26%) 

2 (4%) 

Inflammation, chronic, focal 1 ~(2%) 

Metaplasia, osseous 

Ulcer 

1 (2%) 

(2%) (6%) 

Subcutaneous tissue, inflammation, chronic (2%) 

Musculoskeletal System 

Bone (50) (50) (50) 

Cranium, hyperostosis 1 (2%) 

Cranium, osteopetrosis 1 (2%) 

Femur, osteopetrosis 2 (4%) 1 (2%) 

Nervous System 

Brain 

Compression 

Gliosis 

(50) 

4 (8%) 

(50) 

4 (8%) 

(50) 

2 (4%) 

1 (2%) 

Hemorrhage 

Hydrocephalus 

Mineralization 

Necrosis 

2 (4%) 

1 (2%) 

2 (4%) 

2 (4%) 

1 (2%) 

1 (2%) 

2 (4%) 

Cerebellum, necrosis 1 (2%) 
~ - - ~ _ ~ ~ - . ~ __ ~ 

Dietary Restriction, NTP TR 1 ~ 460 

TABLE A3a 

Summary of the Incidence of Nonneoplasti e Lesions in Male ~ats in the Dietary Restriction Study 

of ~uty~ Benzy~ ]Phtha~ate: Ad Libitum.treeding and Weight-IV~Iatched Controls ]Protocols (continued) 

2- ~rear Study (continued) 

Respiratory System 

Lung 

Congestion 

Hemorrhage 

Inf'fltration cellular, histiocyte 

Inflammation, subacute 

Inflammation, suppurative 

Metaplasia, osseous 

Alveolar epithelium, hyperplasia 

Nose 

Exudate 

Foreign body 

Fungus 

Mucosa, hyperplasia 

Mueosa, metaplasia, squamous 

Special Senses System 

Eye 

Atrophy 

Urinary System 

Kidney 

Inflammation, suppurative 

Mineralization 

Nephropathy 

Renal tubule, cytoplasmic alteration 

Renal tubule, dilatation 

Renal tubule, hyaline droplet 

Renal tubule, necrosis 

Renal tubule, pigmentation 

Transitional epithelium, hyperplasia 

Urinary bladder 

Transitional epithelium, hyperplasia 

Ad LibitumFed Control * 

(50) 

1 (2%) 

13 (26%) 

1 (2%) 

I (2%) 

1 (2%) 

7 (14%) 

(50) 

17 (34%) 

8 (16%) 

12 (24%) 

11 (22%) 

9 (18%) 

(1) 

I (1~%) 

(50) 

48 (96%) 

49 (98%) 

6 (12%) 

(50) 

Weight-Matched 

Control 

(50) 

1 (2%) 

i (2%) 

15 (30%) 

1 (2%) 

1 (2%) 

6 (12%) 

(50) 

11 (22%) 

3 (6%) 

10 (20%) 

8 (16%) 

5 (10%) 

(50) 

1 (2%) 

1 (2%) 

48 (96%) 

2 (4%) 

1 (2%) 

1 (2%) 

49 (98%) 

(50) 

12,000 ppm 

(50) 

1 (2%) 

1 (2%) 

6 (12%) 

2 (4%) 

(50) 

17 (34%) 

5 (10%) 

11 (22%) 

12 (24%) 

2 (4%) 

(1) 

(50) 

4 (8%) 

48 (96%) 

1 (2%) 

1 (2%) 

50 (100%) 

1 (2%) 

(50) 

2 (4%) 
130 Dietary Restriction, NTP TR 460 

TABIJ~ A3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and 30-Month Restricted Feed Protocols a .. 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

Disposition Summary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Accidental death 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

Alimentary System 

Intestine large, colon 

Parasite metazoan 

Intestine large, rectum 

Parasite metazoan 

Liver 

Basophilic focus 

Clear cell focus 

Eosinophilic focus 

Granuloma 

Bile duct, hyperplasia 

Hepatocyte, vacuolization cytoplasmic 

Kupffer cell, pigmentation 

Mesentery 

Fat, necrosis 

Pancreas 

Atrophy 

Cardiovascular System 

Heart 

Cardiomyopathy 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

~. Angiectasis 

Degeneration, fatty 

Hyperplasia, focal 

Pituitary gland 

Pars distalis, cyst 

Pars distalis, hyperplasia, focal 

Thyroid gland 

C-cell, hyperplasia 

60 

10 

1 

14 

1 

34 

(10) 

1 (10%) 

(10) 

1 (10%) 

(10) 

1 (10~) 

1 (10%) 

1 (10%) 

2 (20%) 

(3) 

3 (100%) 

(10) 

4 (40%) 

(10) 

5 (50%) 

(10) 

2 (20%) 

1 (10%) 

2 (20%) 

2 (20%) 

(10) 

2 (20%) 

2 (20%) 

(10) 

1 (10%) 

60 

I0 

50 50 

17 

2 

37 

3 

29 

8 

31 10 13 

50 50 

(10) 

(10) 

2 

(10) 

(2O%) 

3 

1 

1 

1 

(30%) 

00%) 

(10%) 

(10%) 

1 

(1) 

1 

'(10) 

' 5 

(10%) 

(I00%) 

(50%) 

(10) 

4 (40%) 

Oo) 

3 (30%) 

1 (10%) 

~ 00~)

(10) 

~ 00~) 

(1o) 

I (lo%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
131 

--:~ ~ ~ ~.=~- :_ ~:~ ................................ :~. ~,~ 

Dietary Restriction, ~ T~ 460 

T~k~LE A3b 

Summary of the ~nc~dence ot Nonneop~asfic Lesions ~n Male 1Rats ~n the D~e~ry Restriction Study 

o~' ll~uty~ lI~enzy~ PhthMate: 2-Year and 30-Month Restricted Feed ll~'otocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

.~ 5-Month ffnterim Evaluation (continued) 

Genital System 

Epididymis (10) (10) 

Atypia cellular 1 (10%) 1 (10%) 

Preputial gland (10) (10) 

Inflammation, chronic 9 (90%) 10 (100%) 

Inflammation, suppurative 1 (10%) 

Prostate (10) (10) 

Corpora amylacea 2 (20%) 4 (40%) 

Inflammation, suppurative 7 (70%) 4 (40%) 

Epithelium, hyperplasia 1 (10%) 1 (10%) 

Testes (I0) (I0), 

Interstitial cell, hyperplasia 9 (90%) 9 (90%) 

S~miniferous tubule, atrophy 1 (10%) 

Hematopoietic System 

Bone marrow (10) (10) 

Hyperplasia, reticulum cell 1 (10%) 

Myelofibrosis 1 (10%) 

Lymph node (2) 

Mediastinal, hemorrhage 2 (100%) 

Mediastiml, pigmentation 2 (100%) 

Lymph node, mandibular (10) (10) 

Hemorrhage 1 (10%) 1 (10%) 

Pigmentation 3 (30%) 1 (10%) 

Lymph node, mesenteric (10) (10) 

Hyperplasia, lymphoid 1 (10%) 

Spleen (10) (10) 

Pigmentation, hemosiderin 1 (10%) 

Thymus (10) (10) 

Hemorrhage 1 (10%) 1 (10%) 

Respiratory System 

Lung (10) (10) 

Alveolar epithelium, hyperplasia 1 (10%) 

Nose (10) (10) 

Exudate 2 (20%) 

Foreign body 1 (10%) 

Fungus 1 (10%) 

Mueosa, hyperplasia 1 (10%) 

Urinary System 

Kidney (10) (10) 

Mineralization 1 (10%) 

Nephropathy 8 (80%) 10 (100%) 

Renal tubule, pigmentation 8 (80%) 10 (100%) 
132 Dietary Restriction, NTP TR 460 

TABLE A3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2'Year and 30-Month Restricted Feed Protocols (continued) ", 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm: 

15-Month Interim Evaluation (continued) 

Systems Examined With No Lesions Observed 

General Body System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

2-Year and 30-Month Protocols 

Alimentary System 

Intestine large, colon (49) 

Edema 

Parasite metazoan 4 (8%) 

Intestine large, rectum (5o) 

Parasite metazoan 5 (1o%) 

Intestine large, cecum (49) 

Edema 1 (2%) 

Parasite metazoan 1 (2%) 

Ulcer 1 (2%) 

Liver (50) 

Angiectasis " 1 (2%) 

Basophilic focus 16 (32%) 

Clear cell focus 1 (2%) 

Congestion 

Cyst 

Degeneration, cystic 

Developmental malformation 2 (4%) 

Eosinophili c focus 6 (12%) 

Granuloma 1 (2%) 

Hematopnietic cell proliferation 2 (4%) 

Hepatodiaphragmatic nodule 2 (4%) 

Inflammation, subacute 2 (4%) 

Mixed cell focus 1 (2%) 

Thrombosis 

Bile duct, hyperplasia 32 (64%) 

Centrilobular, necrosis 

Hepatucyte, hyperplasia, focal 

Hepatocyte, vacuolization cytoplasmic 4 (8%) 

Kupffer cell, hyperplasia 1 (2%) 

• Kupffer cell, pigmentation 3 (6%) 

Lobules, necrosis, ' 1 (2%) 

Mesentery (7) 

Accessory spleen 2 (29%) 

Fat, necrosis 5 (71%) 

Pancreas (50) 

Atrophy •' ~36 (7~%) ' 

Edema 

Inflammation, chronic 

Acinus, cytoplasmic alteration 2 (4%) 

Acinus, hyperplasia, focal 

(48) 

5 (10%) 

(50) 

5 (10%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

15 (30%) 

5 (10%) 

1 (2%) 

1 (2%) 

4 (8%) 

7 (14%) 

2 (4%) 

1 (2%) 

20 (4O%) 

2 (4%) 

3 (6%) 

(9) 

2 (22%) 

5 (56%) 

(50) 

31 (62%) 

3 (6%) 

(49) (48) 

1 (2%) 

4 (8%) 4 (8%) 

(50) (48) 

5 (10%) 4 (8%) 

(49) (49) 

3 (6%) 8 (16%) 

1 (2%) " 

(50) (49) 

2 (4%) 3 (6%) 

7 (14%) 12 (24%) 

2 (4%) 

1 (2%)• 

1 (2%) 

5 (10%) 1 (2~) 

2' (4%) 

2 (4%) 8 (16%) 

1 (2%) 3 (6%) 

2 (4%)~ 

3 (6%) 2 (4%) 

1 (2%) 

2 (4%) 1 (2%) 

36 ,(72 %) 24 (49%) 

1 (2%) 

1 (2%) 1 (2%) 

2 (4%) 4 (8%) 

5 (10%) 8 (16%) 

5 (10%) 5 (10%) 

(12) (9) 

I (8%) 2 (22%) 

9 (75%) 6 (67%) 

(50) (49) 

30 (6O%) 27 (55%) 

1 (2%) 

1 (2%) 

1 (2%) • 3 (6%) 

2 (4%) 
: .:~-~=~: :~"~,~:~:: ~--~..~..---=::: : :: ..... : . . . . . . . . . . ............................... ,~-----. .... ,. - - . ....... 

Dietary Restriction, ~'P TR 460 ~33 

TABLE A3b 

Summary of the ]Incidence of Nonneoplastie Lesions in Male Rats in the Dietary Restriction Study 

of ~uty~ ~enzy~ IPhtha~ate: 2-Year and 30-Month Restricted Feed lh'otocogs (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

2-Year and 30-Month Protocols (continued) 

Alimentary System (continued) 

Salivary glands (50) 

Atrophy 

Stomach, forestomach (50) 

Edema 1 (2%) 

Hyperplasia, squamous 

Mineralization 

Ulcer 

Mucosa, hyperplasia 1 (2%) 

Stomach, glandular (50) 

Edema 

Erosion 2 (4%) 

Mineralization 

Ulcer 1 (2%) 

Tongue ( 1 ) 

Inflammation, suppurative 

Cardiovascular System 

Heart (50) 

Cardiomyopathy 25 (50%) 

Inflammation, chronic 1 (2 %) 

Thrombosis 1 (2%) 

Endocrine System 

Adrenal cortex (50) 

Accessory adrenal cortical nodule 18 (36%) 

Angiectasis 26 (52%) 

Cyst 

Degeneration, fatty 4 (8%) 

Hematopoietic cell proliferation 1 (2 %) 

Hemorrhage 1 (2%) 

Hyperplasia, focal 9 (18%) 

Hypertrophy, focal 3 (6%) 

Metaplasia, osseous 

Necrosis 

Thrombosis 1 (2%) 

Adrenal medulla (50) 

Hyperplasia 9 (18%) 

Necrosis 

Islets, pancreatic (50) 

Hyperplasia 1 (2%) 

Pituitary gland (46) 

Pars distalis, angiectasis 2 (4%) ~ 

Pars distalis, cyst 6 (13%) 

Pars distalis, hyperplasia, focal 13 (28%) 

Pars intermedia, angiectasis 

Pars intermedia, cyst I (2%) 

(49) 

(50) 

1 (2%) 

1 (2%) 

1 (2%) 

2 (4%) 

3 (6%) 

(50) 

(4%) 

1 (2%) 

(1) 

(50) 

29 (58%) 

1 (2%) 

(50) 

15 (30%) 

26 (52%) 

1 (2%) 

3 (6%) 

2 (4%) 

1 (2%) 

(50) 

8 (16%) 

(50) 

1 (2%) 

(48) 

1 (2%) 

6 (13%) 

6 (13%) 

1 (2%) 

4 (8%) 

(50) 

(50) 

1 (2%) 

2 (4%) 

3 (6%) 

(50) 

2 (4%) 

4 (8%) 

3 (6%) 

(1) 

1 (100%) 

(50) 

30 (6O%) 

1 (2%) 

6 (12%) 

(50) 

15 (30%) 

21 (42%) 

12 (24%) 

1 (2%) 

7 (14%) 

4 (8%) 

3 (6%) 

(50) 

14 (28%) 

1 (2%) 

(50) 

(50) 

2 (4%) 

10 (20%) 

9 (18%) 

6 (12%) 

(50) 

1 (2%) 

(50) 

(49) 

1 (2%) 

6 (12%) 

1 (2%) 

(50) 

32 (64%) 

4 (8%) 

(50) 

18 (36%) 

24 (48%) 

1 (2%) 

4 (8%) ' 

1 (2%) 

• 4 (8%) 

3 (6%) 

1 (2%) 

1 (2%) 

(50) 

4 (8%) 

(49) 

1 (2%) 

(47) 

1 (2%) 

7 (15%) 

lO (12%) 

2 (4%) 
134 Dietary Restriction, NTP TR 460 

TABLE A3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and 30-Month Restricted Feed Protocols (continued) 

 ̧.; 2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

2-Year and 30-Month Protocols (continued) 

Endocrine System (continued) 

Thyroid gland 

C-cell, hyperplasia 

Follicle, cyst 

Follicular ceil, hyperplasia 

(50) 

5 (10%) 

3 (6%) 

(50) 

4 (8%) 

1 (2%) 

(50) • 

8 (16%) 

1 (2%) 

(50) , 

4 (8%) 

1 (2%) 

2 (4%) 

x 

General Body System 

None 

Genital System 

Epididymis (50) (50) (50) (5o) 

Atypia cellular 39 (78%) 35 (70%) 30 (60%) 23 (46%) 

Hypospermia ' 38 (76%) 44 (88%) 39 (78%) 4O (80%)

Preputial gland (50) (50) (50) (50) 

Ectasia 16 (32%) 16 (32%) 14 (28%) 9 (18%) 

Hyperplasia 1 (2%) 1 (2%) 

Inflammation, chi'0nic 26 (52%) 15 (30%) 17 (34%) 13 (26%) 

Inflammation, suppurative 9 (18%) 9 (18%) 12 (24%) 5 (1o%) 

Prostate (49) (50) (50) (50) 

Corpora amylacea 22 (45%) 34 (68%) 22 ~14%) 21 (42%) 

Edema 1 (2%) 

Inflammation, suppurative 22 (45%) . 21 (42%) 23 (46%) 15 (3O%) 

Epithelium, hyperplasia 5 (10%) 8 (16%) 11 (22%) 4 (8%) 

Seminal vesicle (50) . (50) 

" Dilatation ~ I (2%) 

Testes (50) (50) •(50) (5O) 

Interstitial cell, hyperplasia 4 (8%) 3 (6%) 9 (18%) 5 (10%) 

Seminiferous tubule, atrophy 4 (8%) 5 (10%) 5 (10%) 4 (8%) 

Hematopoietic System 

Bone marrow (50) (50) (50) (50) 

Hypercellularity 2 (4%) 2 (4%) 1 (2%) 2 (4%) 

Myelofibrosis 5 (10%) 1 (2%) 4 (8%) 5 (1o%) 

Lymph node (12) (19) (28) (23) 

Deep cervical, hemorrhage 1 0%) 

Iliac, hemorrhage I (4%) 

Inguinal, pigmentation 1 (4%) 

Mediastinal, hemorrhage 1 (8%) 2 (11%) 2 (9%) 

Mediasfi~al,: pigmentation 7 (58%) 4 (21%) 10 (36%) 6 (26%) 

Pancreatic, ectasia 1 (5%) 

Pancreatic, hemorrhage 1 (5%) i (4%) 

Pancreatic, necrosis 1 (4%) 

Pancreatic, pigmentation 4 (33%) 4 (21%) 7 (25%) 5 (22%) 

Renal, pigmentation 1 (8%) 1 (5%) 5 (18%) 2 (9%) 

_. ̧ 
Dietary Restriction, NT~ T~ 460 ~35 

TA~L~ A3b 

Sun, mary o~ the ]Incidence of Honneoplastic Leslons in Male Rats in the Dietary Restriction Study 

of ~uty~ ~en~y9 ]?htha~ate: 2-Year and 30-1V~onth Restricted Feed Frotoco~s (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm 12,000 ppm 

2-3rear and 30-Monel~ Protocols (continued) 

Hematopoietic System (continued) 

Lymph node, mandibular (48) (47) (49) (49) 

Ectasia 5 (10%) 2 (4%) 5 (10%) 2 (4%) 

Hemorrhage 5 ( 10 % ) 7 (15%) 6 (12%) 5 (10%) 

Hyperplasia, lymphoid 2 (4%) 3 (6%) 5 (10%) 1 (2%) 

Pigmentation 10 (21%) 8 (17%) 8 (16%) 6 (12%) 

Lymph node, mesenteric (50) (50) (50) (49) 

Ectasia 2 (4%) I (2%) 

Hemorrhage 2 (4%) I (2%) 2 (4%) 

Hyperplasia, lymphoid 1 (2%) 3 (6%) 1 (2%) 

• Pigmentation 1 (2%) 

Spleen (50) (50) (50) (49) 

Developmental malformation 1 (2 %) 

Fibrosis 7 (14%) 8 (16%) 9 (18%) 15 (31%) 

[ Hematopoietic cell proliferation 5 ( 10 %) 4 (8%) 8 (16%) 4 (8%) 

Inf'dtration cellular, histiocyte 1 (2%) 

Inflammation, chronic I (2%) 

Metaplasia, lipocyte 1 (2%) 

Necrosis I (2%) 1 (2%) 1 (2%) 

~ Pigmentation,hemosiderin 6 (12%) 4 (8%) 8 (16%) 3 (6%) 

Lymphoid follicle, hyperplasia 1 (2 %) 

Thymus (46) (48) (46) (45) 

Eetopic parathyroid gland 1 (2%) 

httegumentary System 

Mammary gland (49) (50) (46) (49) 

Hyperplasia 7 (14%) 10 (20%) 10 (22%) 8 (16%) 

Skin (49) (50) (50) (48) 

Aeanthosis 2 (4%) 2 (4%) 7 (15%) 

Cyst epithelial inclusion 1 (2%) 2 (4%) 1 (2%) 

Hemorrhage 1 (2%) 1, (2%) " 

Hyperkeratosis 2 (4%) 3 (6%) 7 (15%) 

" Inflammation, chronic active 1 (2%) 1 (2%) 3 (6%) 

Ulcer 1 (2%) 2 (4%) I (2%) 

Musculoskeletal System 

Bone (50) (50) (50) (5o) 

Cranium, osteopetrosis 

Femur, osteopetrosis 

2 (4%) 

2 (4%) 

2 (4%) 

2 (4%) 

1 (2%) 

1 (2%) 

Rib, osteopetrosis 1 (2%) 

Nervous System 

Brain (50) (50) (50) (49) 

Compression 2 (4%) 3 (6%) I (2%) 2 (4%) 

Hydrocephalus 

Necrosis 

1 (2%) 

1 (2%) 

3 (6%) 2 (4%) 

2 (4%) 
136 ~ Dietary Restriction, NTP TR 460 

TABLE A3b • 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Studs, 

of Butyl Benzyl Phthalate: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

0 ppm 12,000 ppm 0 ppm • 12,000 ppm . , 

2-Year and 30-Month Protocols (~0ntinued) 

Respiratory System 

Lung (50) (50) (50) (50) 

Congestion 1 (2%) 

• Edema 1 (2%) (2%) • 1 (2%) - Hemorrhage 

Infdtration cellular, histiocyte 4 (8%) ...... 

1 (2%) 

12 (24%) 

(2%) 

(18%) 

, . 

: :12 (24%) 

Inflanunation, subacute I (2%) :2 (4%) 

Inflammation, suppurative 1 (2%) 

Alveolar epithelium, hyperplasia 

Nose 

5 

(50) 

(10%) 2 .(4%) 

(50) 

6 (12%) 

(50) 

.,3 (6%) •

•(50) 

Exudate 5 (10%) 5 (10%) 16 (32%) ,, 7 (14%) 

Foreign body 1 (2%) 8 (16%) 1 (2%) 

Fungus 4 (8%) 3 (6%) I1 (22%) -, 5 (lO%) 

Mucosa, hyperplasia 3 (6%) 1 (2%) 11 (22%) : "6 (12%) 

Mucosa, metaplasia, squamous 3 (6%) '2 (4%) 12 (24%) • 4 (8%) 

sp~ial Senses System 

Eye (1) (I) 

Hemorrhage ~ (ioo%) ' :-

Inflammation, chronic 1 (IOO%) . 1 (100%) 

.., 

Urinary System 

Kidney (50) (5O) (50) (49) 

Calculus, microscopic observation only 1 (2%) 1 (2%) 1 (2%) 

Congestion 1 (2%) 

Cyst 3 (6%) 2 (4%) 

Hydronephrosis 1 (2%) 

Mineralization 5 (10%) 4 (8%) 6 (12%) 5 (10%) 

Nephropathy 43 (86%) 46 (92%) 49 (98%) 48 (98%) 

Thrombosis 1 (2%) 2 (4%) 

Renal tubule, atrophy 3 (6%) 1 (2%) 

" Renal tubule, dilatation 2 (4%) 5 (10%) 2 (4%) 3 (6%) 

Renal tubule, hyperplasia 1 (2%) 

Renal tubule, necrosis " 1 (2%) 1 (2%) 4 (8%) 4 (8%) 

Renal tubule, pigmentation 47 (94%) 47 (94%) 5O (100%) 47 (96%) 

Tramitionai epithelium, hyperplasia 1 (2%) 2 (4%) 2 (4%) 1 (2%) 

Urinary bladder (50) (50) (50) (50) 

Hemorrhage 2 (4%) 1 (2%) 

Transitional epithelium, hyperplasia 1 (2%) 2 (4%) 1 (2%) 

-_ 
< "~::=~ -_~--~ ,=~= _ . . . . . . . . . . . . . . . . . . . . . . . o .............. ~ ....... :~._ ~_=. .... 

_ 

137 

~ ~F~E~~ ~ 

XAVS 

~H T~ ~BTA~ ~ST~CT~ON STaY 

OF BUTYL BEHZYL ~HT~LATE 

TABLE ~a Summary of the Incidence of Neoplasms in Female Rats 

in the Dietary Restriction Study of Butyl Benzyi Phthalate: 

Ad Libitum Feeding and Weight4V~atched Controls Protocols ................... 13~ 

TABLE ~b Summary of the Incidence of Neoplasms in Female Rats 

in the Dietary Restriction Study of Butyl Benzyl Phthaiate: 

2-'Year and 32-IV~onth Restricted Feed Protocols ............................ 142 

~ABLE ~a Statistlcai Analysis of Primary Neoplasms in Female Rats 

in the Dietary Restriction Study of ~uty~ Benzyi Phthaiate: 

Ad Libitum Feeding and Weight-IV~atched Controns ][h-otoco~s ................... ~47 

TABLE ]~2b Statistical Analysis of Primary Neoplasms in Female Rats 

in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

2-Year and 32-1V~onth Restricted Feed Protocols ............................ ~S~ 

TABLE B3a Summary of the Incidence of Nonneoplastic Lesions in Female Rats 

in the Dietary Restriction Study of Butyl Benzy~ Phthalate: 

Ad Libitum Feeding and Weight-lV~atchefl Controls Protocols ............. ...... 156 

TABLE ~3b Summary of the ~ncidence of Nonneoplastic Lesions in Female Rats 

in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

2-Year and 324V~onth Restricted Feed Protocols ............................ 163 

l 
138 Dietary Restriction, NTP TR 460 

TABLE Bla 

Summary of the Incidence of Neoplasms in Female Rats in the Dietary Restriction Study of Butyl Benzyi Phthalate: 

Ad .Libitum Feeding and Weight-Matched Controls Protocols a 

Ad LibitumFed Control 

Disposition Summary 

Animals initially in study 60 

15-Month interim evaluation I0 

Early deaths 

Moribund 23 

-Naturaldeaths 2 

Survivors 

Terminal sacrifice 25 

Animals examined microscopically 60 

t5-Month Interim Evaluation 

Endocrine System 

Pituitary gland (I0) 

Pars distalis, adenoma 

Thyroid gland (10) 

C-cell, adenoma, multiple I (10%) 

Follicular cell, adenoma I (10%) 

Genital System 

Uterus (10) 

-Deciduoma benign 1 (10%) 

Polyp stromal 1 (10%) 

Systemic Lesions 

Multiple organs b . • (10) 

Leukemia mononuclear 1 (10%) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

Weight-Matched ~ 

Control 2A,O00 ppm 

60 60 

I0 10 

7 20 

2 1 

41 29 

60 60 

(10) (I0) 

I (10%) 

(10) (10) 

(10) (I0) 

(10) (1o) 
Dietary ]Restriction, NTP TN 460 ~39 

T~LE ~la 

Surnnaary of the ]Incidence of Neop]asrns~ ~e~nale ~ats ~n the ~etary ]~est~cct~on St~y 

Ad Libitum Feeding and Weight-lV~atched Controls ~Protoco~s (continued) 

o~ ]bury9 ~en~y~ ]~htha~ate: 

: 

Ad Libitum- Weight-Matched 

~ed Control Control ~4,000 ppm 

2-~'ear Study 

Alimentary System 

Intestine large, colon (50) (49) (50) 

Intestine large, cecum (50) (50) (49) 

Intestine small, jejunum (50) (50) (49) 

Carcinoma 1 (2%) 

Intestine small, ileum (49) (49) (50) 

Liver (50) (50) (5O) 

Carcinoma, metastatic, intestine 

small, jejunum 1 (2%) 

Hepatocellular adenoma 2 (4%) 

Mesentery (10) (2) (6) 

Liposarcoma 

Oral mucosa 

1 00%) 

(1) (1) 

Squamous cell papilloma 1 (100%) 

Pancreas (50) (49) (50) 

Acinus, adenoma 2 (4%) 

Salivary glands (49) (50) (50) 

Stomach, forestomach (50) (50) (50) 

Squamous cell papilloma 1 (2%) 

Stomach, glandular 

Tongue 

(50) 

(3) 

(50) 

(1) 

(50) 

Squamous cell papilloma 2 (67%) 1 0OO%) 

Cardiovascular System 

Heart (50) (50) (50) 

Schwannoma malignant 1 (2%) l (2%) 1 (2%) 

Endocrine System 

Adrenal cortex (50) (50) (50) 

Adenoma 

Carcinoma 

I (2%) 

1 (:2%) 

Adrenal medulla 

Pheoehromocytoma complex 

(49) (50) 

1 (2%) 

(50) 

Pheochromoeytoma benign 1 (2%) l (2%) 

Islets, pancreatic (50) (49) (50) 

Adenoma 1 (2%) 

Parathyroid gland (47) (46) (48) 

Adenoma 1 (2%) 

Pituitary gland 

Pars distalis, adenoma 

(49) 

19 (39%) 

(47) 

7 05%) 

(50) 

13 (26%) 

Pars distalis, adenoma, multiple 1 (2%) 

Pars distalis, carcinoma 2 (4%) 

Pars intermedia, adenoma I (2%) 

Thyroid gland (49) (50) (50) 

C-cell, adenoma 4 (8%) 1 (2%) 2 (4%) 

C-cell, carcinoma 1 (2%) 

C-cell, carcinoma, multiple 1 (2%) 

Follicular cell, carcinoma I (2%) 
140 Dietary Restriction, NTP TR 460 

TABLE Bla 

Summary of the Incidence of Neoplasms in Female Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) " " 

2-Year Study (continued) 

General Body System 

Tissue NOS 

Genital System 

Clitoral gland 

Adenoma 

Carcinoma 

Ovary 

Gmnulosa cell tumor benign 

Uterus 

Deciduoma benign 

Lciomyoma 

Lciomyosarcorna 

Polyp stromal 

Hematopoietic System 

Bone marrow 

Lyhaph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

Carcinoma, metastatic, thyroid gland . - • 

Integumentary System 

Mammary gland 

Adenoma 

Carcinoma 

Fibroadenoma 

Skin 

Basal cell adenoma 

Keratoacanthoma 

Trichoepithe!ioma 

Sebaceous gland, carcinoma 

Subcutaneous tissue, fibroma 

Musculoskeletal System 

None 

Nervous System 

Brain 

Astrocytoma malignant 

Carcinoma 

Carcinoma, metastatic, pituitary gland 

Ad LibitumFed Control 

(I) 

(50) 

3 (6%) 

4 (8%). 

(50) 

2 (4%) 

(50) 

I (2%) 

1 (2%) 

6 (12%) 

(50) 

(12) 

(49) 

(50) 

(50) 

(49) 

(49) 

2 (4%) 

2 (4%) 

28 (57%) 

(50) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

2 (4%) 

Weight-Matched 

Control 

' 

(50) 

2 (4%) 

1 (2%) 

(50) 

(50) 

4 (8%) 

(50) 

(13) 

(50) 

(50) 

(50) 

(49) 

(48) 

7 (15%) 

(50) 

1 (2%) 

1 (2%) 

1 (2%) 

I (2%) 

1 (2%) 

: 

(50) 

24,000 ppm 

(I) 

(50) 

4 (8%) 

(50) " 

(50) 

1 (2%) 

7 (14%) 

(50) 

(15) 

(50) 

(50) 

(50) 

(48) 

1 (2%) 

(50) 

I1 (22%) 

(50) 

1 (2%) 

(50) 

1 (2%) 
m :~-~-- ~ :~ "- ~ . . ~ . 

Dietary Restriction, NTP TR 460 141

TA~LI~ ~la 

Sumnmry o~ the ]Incidence of Neo#asms in Female ~ats in the Dietary ]Restriction Study of ~uty~ ~enzyl ~hthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) : , 

Ad ILibitum- Weight-Matched 

Fed Control Control .. 

• , 

2-Year Study (continued) 

Respiratory System 

Lung (50) (50) 

Alveolar/bronchiolar adenoma 2 (4%) 

Special Senses System 

None 

Urinary System 

Kidney (50) (50) 

Hemangioma 

Renal tubule, adenoma 1 (2%) 

Renal tubule, carcinoma 1 (2%) 

Urinary bladder (50) (50) 

Transitional epithelium, papilloma 1 (2%) 

Systemic Lesions 

Multiple organs (50) (50) 

Leukemia mononuclear 21 (42%) 13 (26%) 

Neoplasm Summary 

Total animals with primary neoplasms c 

15-Month interim evaluation 4 1 

2-Year study 49 32 

Total primary neoplasms 

15-Month interim evaluation 5 1 

2-Year study 112 49 

Total animals with benign neoplasms 

15-Month interim evaluation 3 1 

2-Year study 40 19 

Total benign neoplasms 

15-Month interim evaluation 4 1 

2-Year study 77 29 

Total animals with malignant neoplasms 

• 15-Month interim evaluation 1 

2-Year study 30 20 

Total malignant neoplasms 

15-Month interim evaluation 1 

2-Year study 35 20 

Total animals with metastatic neoplasms 

2-Year study 2 

Total metastatic neoplasms 

2-Year study 2 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

e Primary neoplasms: all neoplasms except metastatic neoplasms 

24,000 ppm 

(50) 

(50) 

1 (2%) 

(50) 

2 (4%) 

(50) 

19 (38%) 

/~" 

i 

i:g

:,: 

i 

~: 

~ 

i~ 

0 

42 

i 

70 

31 

48 

22 

22 

1 
142 Dietary Restriction, NTP TR 460 

TABLE Blb 

Summary. of the Incidence of Neoplasms in Female Rats in the Dietary Restriction Study . 

of Butyl Benzyl Phthalate: 2-Year and 32-Month Restricted Feed Protocols a 

2-Year Restricted Feed -~ 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Disposition Summary 

Animals initially in study 

15-Month interim evaluat~n 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

60 

I0 

11 

4 

35 

60 

10 

10 

1 

39 

50 

34 

6 

10 

50 

38 

1 

11 

Animals examined microscopically 60 50 50 

15-Month Interim Evaluation 

Endocrine System 

Pituitary gland 

Pars distalis, adenoma 

(IO) 

1 (10%) 

(1o) 

Genital System 

Clitoral gland 

Adenoma 

~ (10) 

I (I0%) 

(1o) 

Systemic Lesions 

Multiple organs b -

Leukemia mononuclear 

(I0) (1o) 

1 (1o%) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System . 

Urinary System 

2-Year and 32-Month Protocols 

Alimentary System 

Intestine large, colon 

Intestine large~ cecum Intestine small, duodenum 

Intestine small, jejunum 

Leiomyosarcoma 

.. 

(5o) 

(5o) 

(50) 

(50) 

I (2%) 

(49) 

(5o) 

(50) 

(50) 

i (2%) 

(49) 

(50) 

(49) 

(5o) 

(50) 

(50) 

(50) 

(50) 
i 

•.= ~ , ~ : : ~ ~----

Dietary Restriction, ~ TR ~ 1~3 

TABLE II~lb 

Summary of the ]Incidence of Neoplasms in ]~en~a~e Rats ~n the Dietary Restriction Study 

o~ ~uty~ ~enzyi ]?htha~ate: 2-Year and 32-NIonth Restricted Feed ]Protocols (continued) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

2-Year and 32-Month Protocols (continued) 

Alimentary System (continued) 

Intestine small, ileum (50) (50) (50) (50) 

Liver (50) (5O) (50) (50) 

Hepatuceiiniar carcinoma 1 (2%) 2 (4%) 

Hepatucellular adenoma 1 (2 %) I (2%) I (2%) 3 (6%) 

Mesentery (1) (5) (4) 

Lipoma 1 (25%) 

Pancreas (47) (50) (50) (50) 

Acinus, adenoma 1 (2%) 

Salivary glands (49) (50) (50) (50) 

Stomach; forestomach (49) (50) (48) (50) 

Stomach, glandular (50) (50) (48) (50) 

Tongue (1) 

Squamous cell papilloma 1 (100%) 

Tooth (1) (3) 

Odontoma 1 (33%) 

Cardiovascular System 

Heart (50) (50) (50) (50) 

Schwannoma malignant \ i (2%) 

Endocrine System 

Adrenal cortex (50) (50) (50) (50) 

Carcinoma 1 (2%) . : 

Adrenal medulla (49) (50) (49) (50) 

Pbeochromocytoma malignant 1 (2 %) 

Pbeochromoeytoma complex 1 (2 %) 

Pheochromocytoma benign 1 (2 %) 2 (4%) 5 (10%) 

Islets, pancreatic (47) (50) (50) (50) 

Adenoma 1 (2%) i (2%) 

Carcinoma 1 (2%) 

Pituitary gland (50) (49) (50) (50) 

Pars distalis, adenorna 15 (30%) 6 (12%) 23 (46%) 16 (32%) 

Pars distalis, adenoma, multiple 2 (4%) 

Pars distalis, carcinoma 1 (2%) 

Thyroid gland (49) (50) (50) (50) 

C-cell, adenoma 2 (4%) 2 (4%) 4 (8%) 5 (10%) 

C-cell, carcinoma 1 (2%) 

Follicular cell, adenoma 1 (2%) 

Follicular cell, carcinoma 1 (2%) 

General Body System 

Tissue NOS (I) 

Sarcoma 1 (100%) 
144 Dietary Restriction, NTP TR460 

TABLE Blb 

Summary of the Incidence of Neoplasms in Female Rats in the Dietary Restriction Study 

of Butyl Benzyi Phthalate: 2-Year and 32-Month Restricted Feed Protocols (continued) 

.. ' .. 2-Year Restricted Feed ~ 

0 ppm 24,000 ppm 

2-Year and 32-Month Protocols (continued) 

Genital System 

Clitoral gland 

Adenoma 

Carcinoma 

(50) 

3 (6%) 

" 

(50) 

3 (6%) 

Ovary (50) (50) 

Granulosa cell rumor benign 1 (2 %) 

Uterus (50) (50) 

Adenoma 1 (2%) 

Polyp stromal 2 (4%) 

Polyp stromal, multiple 

Sarcoma stromal 

Vagina (I) (1) 

Squamous cell papilloma 

Hematopoietic System 

Bone marrow (50) (50) 

Lymph node (10) (8) 

Lymph node, mandibular (49) (50) 

Lymph node, mesenteric (50) (50) 

Spleen (50) (50) 

Thymus (50) (50 ) 

Thymoma benign 1 (2%) 

Integumentary System 

Mammary gland (50) (49) 

Carcinoma 1 (2%) 

Fibroadenoma 10 (20%) 2 (4%) 

Fibroadenoma, multiple 2 (4%) 

Skin (50) (50) 

Keratoacanthoma 1 (2%) 

Squamous cell papilloma 1 (2%) 

Sebaceous gland, basal cell adenoma 

Subcutaneous tissue, fibroma 1 (2%) 

Subcutaneous tissue, fibroma, multiple 

Subcutaneous tissue, melanoma malignant 

Musculoskeletal System 

Bone (50) (50) 

Osteosarcoma 

Skeletal muscle 

Rhabdomyosarcoma 

32-Month Restricted Feed 

0 ppm 24,000 ppm 

(50) (49) 

5 (10%) 2 (4%) 

5 (10%) 

(50) (50) 

(50) (50) 

3 (6%) 7 (14%) 

I (2%) 

1 (2%) 

(1) (1) 

.1 (100%) 

(50) (50) 

(21) (18) 

(50) (50) 

(50) (50) 

(50) (50) 

(49) (46) 

(49) (50) 

4 (8%) 1 (2%) 

18 (37%) 5(1o%) 

3 (6%) 

(50) (50) 

2 (4%) 1 (2%) 

1 (2%) • 

1 (2%) 

3 (6%) 

I (2%) 

1 (2%) 

(50) (50) 

1 (2%) 

(1) 

1 (100%) 
145 

:%--~~'~-~. ........... -r-~-~-~ .... ---~'77~--"~'~--~7"~'~ ~,~--...~.,~_~ :~ 

Dietary Restriction,.NTP TR 4(~ 

TABLE Blb 

Suanmmry of the ]Incidence of Neoplasms in FemaHe RaN in the Dietary Restriction Study 

of Butyl Benzy~ ~?hthalate: 2-'Year and 32-Month Restricted. Feed Protocols (continued) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

2-Year and 32-Month Protocols (continued) 

Nervous System 

Brain (50) (50) (50) (49) 

Astroeytoma malignant 1 (2 %) 1 (2%) 

Carcinoma, metastatic, pituitary gland 1 (2%) 

Carcinoma, metastatic, Zymbal's gland I (2%) 

Ependymoma malignant 1 (2%) 

Glioma malignant 1 (2%) 

Oligodendroglioma malignant 1 (2 %) (2%) 

Spinal cord (1) (2) (1) 

Respiratory System 

Lung (50) (50) (50) (50) 

Alveolar/bronchiolar adenoma 1 (2 %) 1 (2%) 

Alveolar/bronchiolar carcinoma 1 (2%) 

Carcinoma, metastatic, Zymbal's gland 1 (2%) 

Sarcoma, metastatic, tissue NOS 1 (2%) 

Nose (50) (50) (50) (50) 

Special Senses System 

Zymbal' s gland (1) .(1) (1) (1) 

Carcinoma 1 (100%) 1 (100%) 1 (100%) 1 (100%) 

Urinary System 

Kidney (50) (50) (5O) (50) 

Urinary bladder (50) (50) (49) (50) 

Mast cell rumor benign 1 (2%) 

Transitional epithelium, carcinoma 4 (8%) 

Transitional epithelium, papilloma • 2 (4%) I (2%) 2 (4%) 

Systemic Lesions 

Multiple organs (50) (50) (50) (50) 

Leukemia mononuclear 16 (32 %) 18 (36%) 29 (58%) 39 (78%) 

Mesothelioma NOS I (2%) 
1~6 Dietary Restriction, NTP TR 460 

TABLE Bib 

Summary of the Incidence of Neoplasms in Female Rats in the Dietary .Restriction Study. 

of Butyl Benzyl Phthalate: 2-Year and 32-Month Restricted Feed Protocols (continued) ~ 

2-Year Restricted Feed 32-Month Restricted Feed 

'0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Neoplasm Sunmmry 

Total animals with primary neoplasms ¢ 

15-Month interim evaluation 2 1 

2-Year and 32-month protocols 28 50 49 

Total primary neoplasms 

15-Month interim evaluation 2 1 

2-Year and 32-month protocols 69 41 121 102 

Total animals with benign neoplasms 

15-Month interim evaluation 2 

2-Year and 32-month protocols 29 15 43 34 

Total benign neoplasms 

15-Month interim evaluation 2 

2-Year aml 32-month protocols 18 74 51 

Total animals with malignant neoplasms 

15oMonth interim evaluation 1 

2-Year and 32-month protocols 24 20 39 41 

Total malignant neoplasms 

15-Month interim evaluation 1 

2-Year and 32-month protocols 26 23 47 50 

Total animals with metastatic neoplasms 

' 2-Year and 32-month protocols 1 -1 

Total metastatic neoplasms 

2-Year and 32-month protocols 1 2 

Total animals with uncertain neoplasmsbenign or malignant 

32-Month protocol I 

Total uncertain neoplasms 

32-Month protocol 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

c Prima~ neoplasms: all neoplasms except metastatic neoplasms 
: r ....... ~.- _~_.~:~ . . . . . ~ . _ _ ~ : _ ~ . ~ = = , . . . . . . . . . . . ~,,, _ ............ ..: :--: ~ :~_ :~ :~-

D~e~'y ~estr~ct~oa~ NT]P ~ ~ ~47 

T~LE ~2a 

Stat~sticaH Ana~ys~s o~ ~l~r~aary Neoplasms ~n Female ]Rats in the Dietary ~estr~ction Study o~ ~uty~ ~e~y] Phtha~ate: 

Ad L~b/~um Feeding ~nd Weightq~Iatched Controls l~hrotoco~s 

24,000 ppm × 24,000 ppm × 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Clitoral Gland: Adenoma 

Overall rate a 3/50 (6%) 4/50 (8%) 2/50 (4%) 4/50 (8%) 

Adjusted rateb 11.2% 13.8% 4.9% 13.8% 

Terminal ratec 2/25 (8%) 4•29 (14%) 2/41 (5%) 4/29 (14%) 

First incidence (days) 725 736 (T) 737 (T) 737 (T) 

Life table test d P=0.572 P=0.191 

Logistic regression test d P =0.530 P =0.191 

Fisher exact test d P=0.500 P=0.339 

Clitoral Gland: Carcinoma 

Overall rate 4•50 (8%) 0/50 (0%) 1/5o (2%) 0/50 (o%) 

Adjusted rate 14.3% 0.0% 

Terminal rate 3/25 (12%) 0/29 (0%) 

First incidence (days) 702 _e 

Life table test P=0.054N 

Logistic regression test P=0.069N 

Fisher exact test P =0.059N 

Clitoral Gland: Adenoma or Carcinoma 

Overall rate 7/50 (14%) 4•50 (8%) 3/50 (6%) 4/50 (8%) 

Adjusted rate 24.7% 13.8% 7.3% 13.8% 

Terminal rate 5•25 (20%) 4•29 (14%) 3/41 (7%) 4/29 (14%) 

First incidence (days) 702 736 (T) 737 (T) 737 (T) 

Life table test P=0.198N P=0.315 

Logistic regression test P=0.265N P=0.315 

Fisher exact test P=0.262N P=0.500 

Mammary Gland: Fibroadenoma or Adenoma 

Overall rate 28•50 (56%) 11/50 (22%) 7/50 (14%) II/50 (22%) 

Adjusted rate 71.0% 28.9% 16.5% 28.9% 

Terminal rate 14/25 (56%) 5/29 (17%) 6/41 (15%) 5/29 (17%) 

First incidence (days) 587 487 722 487 

Life table test P=0.001N P=0.084 

Logistic regression test P < 0.00IN P=0.225 

Fisher exact test P < 0.001N F=0.218 

Mammary Gland: Adenoma or Carcinoma 

Overall rate 4/50 (8%) 0/50 (0%) 0/50 (0%) 0/50 (0%) 

Adjusted rate 14.6% 0.0% 

Terminal rate 3/25 (12%) 0/29 (0%) 

First incidence (days) 716 -

Life table test P=0.052N 

Logistic regression test P=0.064N 

Fisher exact test P=0.059N 

l 
148 Dietary Restriction, NTP TR 460 

TABLE B2a 

Statistical Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) :., 

24,000 ppm x 24,000 ppm × 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control. Control 

Mammary Gland: Fibroadenoma, Adenoma, or Carcinoma 

Overall rate 29/50 (58%) 

Adjusted rate 71.8 % 

Terminal rate 14/25 (56%) 

First incidence (days) 587 

Life table test 

Logistic regression test 

Fisher exact test : 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 20/49 (41%) 

Adjusted rate. 57.0% 

Terminal rrate 11/25 (44%). 

First incidence (days) 598 

Life table test 

Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): Adenoma or Carcinoma 

Overall rate 22/49 (45 %) 

Adjusted rate 61.3% 

Terminal rate • 12/25 (48%) 

First incidence (days) . 598 

Life table test . 

" 

Logistic regression test : 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma 

Overall rate 4/49 (8 % ) 

Adjusted rate 11.4% 

Terminal rate 1/25 (4%) 

First incidence (days) 702 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 5/49 (10%) 

Adjusted rate 15.1% 

Terminal rate 2/25 (8%) 

First incidence (days) 702 

Life table test 

Logistic regression test 

Fisher exact test 

11/50 (22%) 

28.9 % 

5/29 (17,%) 

487 

P<0.001N 

P < 0.001N 

P <0.001N 

13/50 (26%) 

36.1% 

7/29 (24%) 

592 

P=0.086N 

P=0.121N 

P=0.088N 

13/50 (26%) 

36.1% 

7/29 (24%) 

592 

P=0.042N 

P=0.058N 

P=0.039N 

2/50 (4 % ) 

6.9% 

2/29 (7%) 

736 (T) 

P=0.326N 

P=0.359N 

P=0.329N 

3/50 (6 %) 

8.8% 

2/29 (7%) 

452 

P=0.337N 

P=0.346N 

P=0.346N 

7/50 (14%) 

16.5 % 

•6/41 (15%) 

722 

7/47 (15%) 

17.1%, 

5/38 (13%) 

722 

7/47 (15 %) 

17.1% 

5/38 (13%) • 

722 

1/50 (2 %) • 

1/50 (2%) 

2.4% 

1/41 (2%) 

737 (T) 

. ,: 

11/50 (22%) 

28.9% 

5/29 (17%) 

487 

P=0,084 

P--0.225 

P=0.218 

13/50 (26%) 

36.1% 

7/29 (24 %) 

592 

P=0.033 

P=0.077 

P=0.135 

13/50 (26%) 

36.1% 

7/29 (24%) 

592 

P=0.033 

P=0.077 

P=0.135 

2/50 (4%) 

3/50 (6%) 

8.8% 

2/29 (7%) 

452 

P=0.219 

P=0.336 

P=0.309 
:~:: ~S==~..: : 7_.~--~ ~ .~,__~ . ...... 7- ................... ;~--~..~ ....... ~ . -~ ..... ~-:~--~, \ . . . . ~--~-~--~__~ ................ ~,_ 

Dietary Restriction, NT~ TR ~60 ~49 

TABLE ~2a 

S~t~s~fc~d An~Kysfs of IPrhnzry NeopKm~ns fin Female Rats fn ~he .Dietary ~es~rfc~tfon S~udly o~ ll~u~yK lI~en~,~ 1Ph~h~a~e: 

Ad Libitum ~eedfng ~nd Wefgh~-Ma~ched Controls Pro~toco~s (continued) 

24,000 ppm. x 24,~00 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

I Uterus: Stromal ~olyp 

Overall rate 6/50 (12%) 7/50 (14%) 4/50 (8%) 7•50 (14%) 

Adjusted rate 19.6% 22.4% 9.8% 22.4% 

Terminal rate 3/25 (12%) 6/29 (21%) 4/41 (10%) 6•29 (21%) 

First incidence (days) 681 487 737 (T) 487 

i Life table test P=0.571 P=0.111 

, Logistic regression test P=0.470 P=0.214 

Fisher exact test P=0.500 P=0.262 

I All Organs: Mononuelear Cell Leukemia 

I Overall rate 21/50 (42%) 19/50 (38%) 13/50 (26%) 19/50 (38%) 

1 Adjustedrate 51.7% 46.1% 28.6% 46.1% • 

Terminal rate 7/25 (28%) 8/29 (28%) 9/41 (22%) 8•29 (28%) 

First incidence (days) 368 452 551 452 

Life table test P=0.398N P=0.034 

] Logistic regression test P=0.422N P=0.302 

'i Fisher exact test P=0.419N P=0.142 

All Organs: Benign Neoplasms 

Overall rate 40/50 (80%) 31/50 (62%) 19/50 (38%) 31/50 (62%) 

Adjusted rate 88.7 % 74.9 % 43.2 % 74.9 % 

Terminal rate 20/25 (80%) 19/29 (66%) 16/41 (39%) 19/29 (66%) 

First incidence (days) 512 452 722 452 

Life table test P=0.060N P<O.001 

Logistic regression test P=O.047N P=O.006 

Fisher exact test P=O:O38N P=O.O14 

All Organs: Malignant Neoplasms 

Overall rate 30/50 (60%) 22/50 (44%) 20/50 (40%) 22/50 (44.%) 

Adjusted rate 70.1% 50.7% 42.3% 50.7% 

Terminal rate 13/25 (52%) 9/29 (31%) 14/41 (34%) 9/29 (31%) 

First incidence (days) 39 404 225 404 

Life table test P=0.110N P=0.128 

Logistic regression test P=0.067N P=0.582 

Fisher exact test P=O.080N P=0.420 
150 Dietary Restriction, NTP TR 460 

TABLE B2a 

Statistical Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of Butyl Benzyl. Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

24,000 ppm X 24,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control . Control Control 

All Organs: Benign or Malignant Neoplasms 

Overall rate 49•50 (98 %) 42•50 (84 %) 32/50 (64 %) 42•50 (84 %) 

Adjusted rate 98.0% " 84.0% 65.3% 84.0% 

Terminal rate 24/25 (96%) 21/29 (72%) 24/41 (59%) 21/29 (72%) 

First incidence. (days) 39 404 225 404 

Life table test t P =0.123N P=0.002 

Logistic regression te~t P=0.027N P=0.054 

Fisher exact test P=0.015N P=0.020 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of anim61s examined microscopically for clitoral gland, 

pituitary gland, thyroid gland, and uterus; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of~he study after adjustment for intercurrent mortality 

c Observed incidence at terminal kill 

O Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the ad libitum-fed or weight-matched controls and the 

exposed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in an 

exposure group is indicated hy N. 

e Not applicable; no neoplasms in animal group 
-- 

~ , ~ :;~ :,~ .... ~_ : '. 

Dietary Restriction, NTP TR 460 151 

TABLE B2b 

Stafistlcal Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of ~uty~ ~euzyl Phthalate: 

2-Year and 32-Month Restricted Feed Protocols 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Adrenal Medulla: Benign Pheochromocytoma 

Overall rate a 1/49 (2%) 0/50 (0%) 

Adjusted rate b 

Terminal rate c 

First incidence (days) 

Life table test d 

Logistic regression test d 

Fisher exact test d 

Adrenal Medulla: Benign, Complex, or Malignant Pheochromocytoma 

Overall rate 3/49 (6%) 0/50 (0%) 

Adjusted rate 7.9% 0.0% 

Terminal rate 2/35 (6%) 0/39 (0%) 

First incidence (days) 624 -

Life table test P=0.108N 

Logistic regression test P=0.118N 

Fisher exact test P=0.117N 

Clitoral Gland: Adenoma 

Overall rate 3/50 (6%) 3/50 (6%) 

Adjusted rate 16.4% 8.8% 

Terminal rate 2/35 (6%) 3/39 (8%) 

First incidence (days) 677 731 (T) 

Life table test P=0.616N 

Logistic regression test P=0.636N 

Fisher exact test P=0.661N 

Clitoral Gland: Carcinoma 

Overall rate 0/50 (0%) 0/50 (O%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Clitoral Gland: Adenoma or Carcinoma 

Overall rate 3/50 (6%) 3•50 (6%) 

Adjusted rate 16.4% 8.8% 

Terminal rate 2/35 (6%) 3/39 (8%) 

First incidence (days) 677 731 (T) 

Life table test P=0.616N 

Logistic regression test P=0.636N 

Fisher exact test P=0.661N 

2/49 (4%) 

15.6% 

1/10 (10%) 

922 

2/49 (4%) 

15.6% 

1/10 (10%) 

922 

5/50 (10%) 

16.2% 

0/10 (0%) 

649 

5/50 (10%) 

38.5 % 

3/10 (30%) 

829 

10/50 (20%) 

48.5% 

3/10 (30%) 

649 

5/50 (1o%) 

18.8% 

1/11 (9%) 

716 

P=0.296 

P=0.244 

P=0.226 

5/50 (10%) 

18.8% 

1/11 (9%) 

716 

P=0.296 

P=0.244 

P=0.226 

2/49 (4%) 

11.7% 

1/11 (9%) 

779 

P~0.176N 

P=0.253N 

P=0.226N 

0/49 (0%) 

0.0% 

0/11 (0%) 

P=0.023N 

P=0.021N 

P=0.030N 

2/49 (4%) 

11.7% 

1/11 (9%) 

779 

P=0.010N 

P=0.012N 

P=0.015N 
152 Dietary Restriction, NTP TR 460 

TABtJ~ B2b 

Statistical Analysis .of, Primary Neoplasms in Female Rats in the Dietary Restriction Study of Butyl Benzyl :Phthalate: 

2-Year and 32,Month Restricted Feed Protocols (continued) ! ,, , .... 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Liver: Hepatocellular Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) : 

Life table test • ' 

Logistic regression test 

Fisher exact test 

,-

,: 

1/50 (2%) I150 (2%) 

: 

1/50 (2%) 

10.0% 

1/10(10%) 

977 (T) 

3/50 (6%) 

20.0% 

2/11 (18%) 

696 

P=0.344 

P=0.348 

P=0.309 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rate 

Adjusted rate " 

Terminal rate 

First incidence (days) 

Life table test : 

Logistic regression test 

Fisher exact test 

1/50 (2%) 2/50 (4%) I150 (2%) 

10.0% 

1/10 (10%) 

977 (T) 

5/50 (10%) 

29.5% 

2/11 (18%) 

696 

P=0.136 

P=0.131 

P=0.102 

Mammary Gland: Fibroadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) ' ' 

Life table test 

Logistic regression test . 

Fisher exact test 

12/50 (24%) 

~, 44.9% 

12/35 (34%) 

730 (T) 

2/50 (4%) 

5.0% 

1/39 (3%) 

600 

P=0.002N 

P=0.003N 

P=0.004N 

21/50 (~2%i 

78.9% 

5/10 (50%) 

687 

7/50 (10%) 

26.1% 

2/11 (18%) 

796 

P<0.001N 

P<0.001N 

P<0.001N 

Mamm~try Gland: Carcinoma 

Overall rate 

Adjusted rate. 

Terminal rate ~ 

First incidence (days) 

Life table test -

Logistic ~egression te~t 

Fisher exact test , 

1/50 (2%) 0/50 (O%) 4/50 (8%) 

14.8% 

0/10 (0%) 

694 

1/50 (2%) 

9.1% 

1/11 (9%) 

977 (T) 

P=0.150N 

P=0.173N 

P=0.181N 

Mammary Gland: 

Overall rate 

Adjusted rate 

Terminal rate • 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Fibr0adenoma or Carcinoma 

13/50 (26%) 

46.4% 

12/35 (34%) 

•. 712 

2/50 (4%) 

5.0% 

1/39 (3%) 

600 

P=0.001N 

P=0.002N 

P=0.002N 

- 24/50 (48%) 

81.5% ' 

5/10 (50%) 

687 

6/50 (12%) 

34,4% 

3/11 (27%) 

796 

v<0.ooIN 

P<0.001N, 

P<0.001N 
-- 

~, :'~..r..:::__. :-=--"7"~ . __~:. ............................ ~ . . . . . . . . . . . , ~ ~ ........ ~ 

Dietary Restriction, NTP TR 460 153

TABLE ~2b 

Statistical Analysis of ]Primary NeopHasrns in ~ernale ~ats in the Dietary ~estriction Stuc~y o~ ~uty~ ~enzy~ Phthalate: 

2-Year anO 32-Month ~estrlcted Feed Protocols (continued) .~ 

2-Year Restricted Feed . 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

F 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 15/50 (30%) 

Adjusted rate 45.8% 

Terminal rate 12/35 (34%) 

First incidence (days) 564 

Life table test I Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): Adenoma or Carcinoma 

Overall rate 15/50 (30%) 

Adjusted rate 45.8 % 

Terminal rate 12135 (34%) 

First incidence (days) 564 

Life table test I 

== ̧! I Logistic regression test 

Fisher exact test 

Skin: Squamous Cell Papilloma or Keratoacanthoma 

Overall rate , , 2/50 (4%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): Fibroma 

Overall rate 1/50 (2%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma 

Overall rate 2/49 (4%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

6/49 (12%) 

14.9% 

4/39 (10%) 

600 

P=O.OI6N 

P=O.O23N 

P=O.027N 

6/49 (12%) 

14.9% 

4/39 00%) 

6OO 

P=0.016N 

P=0.023N 

P=0.027N 

0/50 (0%) 

0/50 (0%) 

2/50 (4%) 

25/50 (50%) 

85.4% 

7/10 (70%) 

478 

26•50 (52 %) 

86.0% 

7/10 (70%) 

478 

3/50 (6%) 

23.1% 

2/10 (20%) 

816 

4/50 (8%) 

28~0% 

2110 (20%) 

687 

4/50 (8%) 

18.4% 

1/10 (10%) 

722 

., , 

16/50 (32%) 

81.8% 

8/11 (73%) 

731 

P=0.O28N 

P=0.018N 

P=0.O52N 

16/50 (32%) 

81.8% 

8/11 (73%) 

731 

Pf0.OI9N 

P=0.011N 

P=0.O34N 

1/50 (2%) 

9.1% 

1/11 (9%) 

977 (rr) 

P=O.261N 

P=O.256N 

P=O.309N 

0/50 (0%) 

0.0% 

0/11 (0%) 

P=0.054N 

P=0.048N 

P=0.059N 

5/50 (10%) 

20.4% 

0/11 (0%) 

634 

P=0.591 

P=0.491 

P=0.500 

[ 
-- 

154 Dietary Restriction, NTP TR 460 

TABLE B2b 

Statistical Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

2-Year and 32-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 2/49 (4%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

) 

Urinary Bladder: Carcinoma 

Overall rate 0/50 (0%) 

Adjusted rate 

Terminal rate 

First incidence (day§) 

Life table test 

Logistic regression test 

Fisher exact~test 

Urinary Bladder: Papilloma or Carcinoma 

Overall rate 0/50 (0%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test ' 

Uierus: Str0mal Polyp or Stromal Sarcoma 

Overall rate 0/50 (0%) 

Adjusted rate 0.0% 

Terminal rate 0/35 (0%) 

First incidence (days) - -

Life table test 

Logistic regression test 

Fisher exact test • • 

All Organs: Mononuclear Cell Leukemia 

Overall rate 16/50 (32%) 

Adjusted rate 43.9% 

Terminal rate 7/35 (20%) 

First incidence (days) 359 

Life table test 

Logistic regression test 

Fisher exact test 

2/50 (4%) 

0/50 (0%) 

2/50 0%) 

2/50 (4%) 

18/50 (36%) 

40.8% 

11/39 (28%) 

592 

P=0.530 

P=0.375 

P=0.417 

5/50 (10%) 

24.7% 

1/10 (10%) 

722 

0/49 (0%) 

0.0% 

0/10 (0%) 

1/49 (2%) 

2.6% 

0/10 (0%) 

694 

4/50 (8%) 

24.0% 

2/10 (20%) 

649 

29/50 (58 %) 

84.6% 

6/10 (60%) 

358 

5/50 (10%) 

20.4% 

0/11 (0%) 

634 

P=0.535N 

P=0.620N 

P=0.630N 

4/50 (8%) 

29.0% 

3/11 (27%) 

719 

P=0.075 

P=0.079 

P=0.061 

6/50 (12%) 

41.0% 

4/11 (36%) 

719 

P=0.077 

P=0.077 

P=0.059 

7/50 (14%) 

32.6% 

2/11 (18%) 

684 

P=0.345 

P=0.289 

P=0.262 

39/50 (78%) 

.91.7% 

8/11 (73%) 

530 

P=0.288 

P=0.027 

P=0.026 
1 

Dietary Restriction, NT~ T~ 46~ ~55 

TABLE S2b 

Stat~st~ca~ Analysis of PHmary Neopaasms ~n Female Rats ~n t h e D i e t a r y ~ e s t H c t ~ o n S t u d y of ~utynBenzyH Phthalate: 

2-Year a~d 32-I~onth Restricted Feed Protocols (continued) .~ 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

All Organs: Benign Neoplasms 

Overall rate 29/50 (58%) 15/50 (30%) 43/50 (86%) 34/50 (68%) 

Adjusted rate 84.9% 38.4% 100.0% 96.8% 

Terminal rate 24•35 (69%) 12/39 (31%) 10/10 (100%) 10/11 (91%) 

First incidence (days) 564 600 478 634 

Life table test P=0.002N P=0.043N 

Logistic regression test P=0.002N P=0.002N 

Fisher exact test P=0.004N P=0,028N 

All Organs: Malignant Neoplasms 

Overall rate 24/50 (48%) 20/50 (40%) 39/50 (78%) 41/50 (82%) 

Adjusted rate 58.2% 44.3% 97.0% 94.7% 

Terminal rate • 11/35 (31%) 12/39 (31%) 9/10 (90%) 9/11 (82%) 

First incidence (days) 359 498 358 530 

Life table test P=0.215N P=0.382N 

Logistic regression test P=0.304N P=0.426 

Fisher exact test P=0.273N P=0.402 

All Organs: Benign or Malignant Neoplasms 

Overall rate 42/50 (84%) 28/50 (56%) 50/50 (100%) 49/50 (98%) 

Adjusted rate 92.1% 58.9% 100.0% 100.0% 

Terminal rate 28/35 (80%) 18/39 (46%) 10/10 (100%) 11/11 (100%) 

First incidence (days) 359 . 498 358 530 

Life table test P=O.006N P=O.226N 

Logistic regression test P=0.001N P=0.404N 

Fisher exact test P=0.002N P=0.500N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, clitoral 

gland, liver, pituitary gland, thyroid gland, urinary bladder, and uterus; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

e Observed incidence at terminal kill 

d Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the controls and that exposed group. The life table 

test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in an exposure group is indicated by 

N. 

e Not applicable; no neoplasms in animal group 
156 Dietary Restriction, .NTP TR460 

TABLE B3a 

Summary of the Incidence of Nonneoplastie Lesions in Female Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocol# 

Disposition Summary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

Alimentary System " 

Intestine large, colon 

Parasite metazoan . 

Intestine large, rectum 

Parasite metazoan 

Liver 

Basophilic focus 

Eosinophilic focus - . 

Granuloma 

Hepatodiaphragmatic nodule 

Bile duct, hyperplasia 

Mesentery 

Fat, necrosis 

Pancreas 

Atrophy 

Tongue 

Epithelium, hyperplasia 

cardiovascular System 

Heart 

Cardiomyopathy 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Angiectasis 

Cyst . 

Degeneration, fatty 

Hyperplasia, focal 

Hypertrophy, focal 

Ad Libitum- Weight-Matched 

Fed Control COntrol 24,000 ppm 

60 60 60 

10 10 10 

23 7, 20 

2 2 1 

25 41 29 

60 

(10) (10) (10) 

I (10%) 1 (I0%) 

(10) (8) (10) 

2 (25%) 

(10) (10) (10) 

l0 000%) 7 (70%) 9 (90%) 

1 (10%) 

1 (10%) 2 (20%) 

1 00%). 3 (30%) 

1 (10%) 

O)

1 (100%) 

(10) (lO) (I0) 

1 (10%) 1 (lO%) 

(I) 

1 (100%) 

., 

(1o) (1o) (10) 

i (1o%) 

,/ 

(lO) (10) (10) 

2 (20~) 4 (40%) 4 (40%) 

5 (50%) ~ (10%) 3 (3o9~) 

1 (lO%) 

1 (10%) 

~ (lO%) 1 00%) 

1 (10%) ~., 

a Number of animals examined microscopically at the site and the n~mber of animals with lesion 
l 

" " ......... ~ =~ ....... '~ ~- -~7 ~ ::~ :~,~-~ ......... -~-~,v~ ~,,, ...... 

D~e~ry Restriction, ~ T]~ ~ 

T~a~L~ ~3a 

S~mary o~' the ]Incidence of Honneop~astic Lesions in Fe~na~e Rats in the Dietary. Restriction Study. 

of ~uty~ ~en~y~ ]Phtha~ate: Ad Libitum Feeding and We~ght-Matche~ Controls Protocols (continued): .: 

]157 

Ad Libitum- Wei~tt-Matched 

Fed Control Control 

~ 5-MOnf~ ~le~m Evalu~on (continued) 

Endocrine System (continued) 

Pituitary gland 

Pars distalis, angiectasis 

Pars distalis, cyst 

Pars distalis, hyperplasia, focal 

Thyroid gland 

Ultimobranchial cyst 

Genital System 

Clitoral gland 

Ectasia 

Inflammation, chronic 

Ovary 

Cyst 

Uterus 

Hydrometra 

Hyperplasia, cystic 

Inflammation, suppurative 

Hematopoietic System 

Bone marrow 

Myelofibrosis 

Lymph node 

Mediastinal, hemorrhage 

Mediastinal, pigmentation. 

Lymph node, mandibular 

Ectasia 

Hemorrhage 

Hyperplasia, lymphoid 

Pigmentation 

Spleen 

Hematopoietic cell proliferation 

Pigmentation, hcmosiderin 

Integumentary System ' 

Mammary gland 

Hyperplasia 

Musculoskeletal System 

Bone 

Cranium, osteopetrosis 

Femur, osteopetrosis 

(10) 

I (10%) 

6 (60%) 

I (10%) 

(10) 

I (10%) 

(I0) 

2 (20%) 

(1o) 

(1o) 

2 (2o%) 

1 (10%) 

(lO) 

(10) 

1 (10%) 

2 (20%) 

3 (30%) 

(10) 

1 (10%) 

10 (100%) 

(lO) 

3 (30%) 

(10) 

(lO) 

1 (1o%) 

2 (20%) 

1 (10%) 

(10) 

(1o) 

(I0) 

(lO) 

1 (10%) 

(lO) 

1 (10%) 

(lO) 

2 (20%) 

(10) 

1 (10%) 

10 (100%) 

(10) 

(1o) 

2~,000 ppm 

(lO) 

1 (10%) i

1 (10%) [ 

(10) 

i = 

(10) 

1 (10%) 

(10) 

1 (10%) 

(10) 

2 (20%) 

1 (10%) 

(10) 

(i) 

1 (100%) 

1 (100%) 

(io) 

2 (20%) 

1 (10%) 

4 (4O%) 

(lO) 

3 (30%) 

lO (100%) . • 

(lO) 

I (10%) 

(10) 

~ (10%) 

2 (20%) 
158 Dietary Restriction, NTP TR 460 

TABLE B3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocols (cbntinued) 

Ad LibitumFed Control 

15-Month Interim Evaluation (continued) 

Respiratory System 

Lung (10) 

Infiltration cellular, histiocyte 2 (20 %) 

Nose (!0) 

Exudate 1 (10%) 

Fungus 1 (10%) 

Mucosa, metaplasia~ squamous I (10%) 

Urinary System 

Kidney ,. (10) 

Cyst 1 (10%) 

Mineralization 10 (100%) 

Nephropa~y 7 (70%) 

Renal tubule, atrophy 1 (10%) 

Renal tubule, pigmentation 10 (100%) 

Systems Examined With No Lesions Observed 

General Body System 

Nervous System 

Special Senses System 

2- Year Study 

Alimentary System 

Intestine large,, colon (50) 

Parasite metazoan 3 (6%) 

Intestine large, rectum (50) 

Parasite metazoan 3 (6%) 

Intestine large, cecum (50) 

Edema 2 (4%) 

Parasite me~azoan 2 (4%) 

Intestine small,, ileum (49) 

Ulcer 1 (2%) 

Liver (50) 

Basophilic focus 37 (74%) 

Clear cell focus 3 (6%) 

Cyst 

Developmental malformation 1 (2%) 

Eosinophilic focus 19 (38%) 

Granuloma 6 (12%) 

Hematopoietic cell proliferation 1 (2%) 

Hepatodiaphragmatic nodule . 9 (18%) 

Inflammation, subacute 3 (6%) 

Mixed cell focus 10 (20%) 

Bile duct, hyperplasia 10 (20%) 

Weight-Matched 

Control 

(10) 

3 (30%) 

(lo) 

(10) 

9 (90%) 

2 (20%) 

9 (90%) 

(49) 

1 (2%) 

(50) 

4 (8%) 

(50) 

! (2%) 

(49) 

(50) 

40 (80%) 

2 (4%) 

16 (32%) 

4 (8%) 

2 (4%) 

4 (8%) 

1 (2%) 

4 (8%) 

10 (20%) 

24,000 ppm 

(10) 

2 (20%) 

(lO) 

(10) 

8 (80%) 

10 (100%) 

IO (100%) 

(50) 

3 (6%) 

(5O) 

4 (8%) 

(49) 

1 (2%) 

! (2%) 

(50) 

(50) 

39 (78%) 

9 (18%) 

2 (4%) 

20 (40%) 

2 (4%) 

1 (2%) 

12 (24%) 

6 02%) 

9 (18%) 
159 

:!=: :~-~r~w~::_~-~-= ....... ~.________. ............ ~ ........ , .............. : : ~ . ~ , ~ - - ~ _ ~ __., ~ . . . . . . . . . . 

Die~ry Restriction, NTP 'TR 4~ 

TABLE B3~ 

Suennaary ol~ the ]Inci~er~ce of Nonne~p~astic Lesions in Fen~e~e Ream h~ ~the ]]~e~.e~'y Res~tr~cction S~tu~y 

of ~uty~ ~enzy~ Ph~ha~ate: Ad Libit~tm ]Feeding ~n~ Weigh~t-IV~che~9 Controls ]Pro~o~:oOs (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 2.4,~1~0 ppm 

2-Year Study (continued) 

Alimentary System (continued) 

Liver (continued) (50) (50) (50) 

Centrilobular. necrosis 1 (2%) 2 (4%) 

Hepatocyte. vacuolization cytoplasmic 7 (14 % ) 1 (2%) 

Kupffer cell. hypcrplasia 1 (2%) 

Kupffer ceil. pigmentation 4 (8%) 4 (8%) 10 (20%) 

Lobules. necrosis 6 (12%) 1 (2%) 6 (12%) 

Mesentery (10) (2) (6) 

Accessory spleen 1 (10%) 1 (17%) 

Fat, necrosis 9 (90%) 2 (100%) 6 (I00%) 

Oral mucosa (1) (I) 

Hyperplasia 1 00O%) 

Pancreas (50) (49) (5O) 

Atrophy 18 (36%) 17 (35%) 12 (24%) 

Cyst 1 (2%) 

Aeinus, cytoplasmic alteration 1 (2%) 2 (4%) 

Acinus, hyperplasia, focal 1 (2%) 

Salivary glands (49) (50) (50). 

Atrophy 1 (2%) 

Hyperplasia 1 (2%) 

Stomach, forestomaeh (50) (50) (50) 

Edema 2 (4%) 

Ulcer 1 (2%) 1 (2%) 

Mucosa, hyperplasia 1 (2%) 1 (2%) 

Stomach, glandular (50) (50) (5O) 

Erosion 1 (2%) 

Inflammation, chronic 1 (2%) 

Mineralization 1 (2%) 

Ulcer 3 (6%) 

Tongue (3) (1) 

Epithelium, hyperplasia 1 (33%) 

Cardiovascular System 

Heart (50) (50) (50) 

Cardiomyopathy 16 (32%) 14 (28%) " 13 (26%) 

Mineralization 1 (2%) 

Thrombosis 1 (2%) 1 (2%) 

Endocrine System 

Adrenal cortex (50) (50) (50) 

Accessory adrenal cortical nodule 5 (10%) 6 (12%) 6 (12%) 

Angiectasis 22 (44%) 15 (30%) 29 (58%) 

Cyst 1 (2%) 

Degeneration, fatty 10 (20%) 4 (8%) 4 (8%) 

Hemorrhage 1 (2%) 

Hyperplasia, focal 9 (18%) 9 (18%) 2 (4%) 

Hypertrophy, focal 9 (18%) 3 (6%) 

Metaplasia, osseous 1 (2%) 
160 Dietary Restriction, NTP TR 460 

TABLE B3a 

Summary of the Incidence' of Nonneopl~tstic Lesions in Female Rats in the Dietary Restriction Study 

of Butyl BenzYl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocols(eonfinu.ed) ' 

Ad Libitum- Weight-Matched 

Fed Control Control 24,000 ppm 

2- Year Study (continued) 

Endocrine System (continued) 

Adrenal medulla (49) (50) (50) 

Hyperplasia 1 (2%) 3 (6%) 1 (2%) 

Islets, pancreatic (50) (49) (50) • Metaplasia, hepatocyte 1 (2%) 

Pituitary gland (49) (47) (50) 

Pars distalis, angiectasis 7 (14%) 7 (15%) 7 (14%) 

Pars distalis, cyst 24 (49%) 8 (17%) 19 (38%) 

Pars distalis, hyperplasia 1 (2%) 

Pars distalis, hyperplasia, focal 12 (24%) 13 (28%) 11 (22%) 

Pars intermedia, cyst 2 (4%) 

Pars intermedia, hyperplasia 1 (2%) 

Thyroid gland (49) (50) (50) 

Ultimobranchial cyst 2 (4%) 

C-cell, hyperplasia 6 (12%) 4 (8%) (6%) 

Follicular cell, hyperplasia (2%) 

General Body System 

Tissue NOS (1) (I) 

Hemorrhage 1 (100%) 

Genital System 

Clitoral gland (50) (50) (5o) 

Ectasia 12 (24%) 4 (8%) 10 (20%) 

Hyperplasia 1 (2%) 1 (2%) 1 (2%) 

Inflammation, chronic 4 (8%) 3 (6%) 1 (2%) 

Inflammation, suppurative 3 (6%) 4 (8%) 3 (6%) 

Ovary (50) (50) (50) 

Cyst 3 (6%) 5 (10%) 

Inflammation, chronic 1 (2%) 

Uterus (50) (50) (5o) 

Hydrometra 2 (4%) 4 (8%) 3 (6%) 

Hyperplasia, cystic 4 (8%) 4 (8%) 11 (22%) 

. 

Hematopoietic System 

Bone marrow (50) (5O) (50) 

Hypercellularity 1 (2%) 2 (4%) 1 (2%i 

Myelofibrosis 4 (8%) 2 (4%) 6 (12%) 

Lymph node (12) (13) (15) 

Mediastinal, hemorrhage 1 (8%) 3 (23%) 1 (7%) 

Mediastinal, pigmentation 2 (17%) 6 (46%) 9 (60%) 

Pancreatic, pigmentation 1 (8%) 3 (20%) 

Renal, hemorrhage 1 (8%) 

Renal, pigmentation 1 (8%) 4 (31%) 4 (27%) 
i ............. ~ ~"r ~$~4 ~ 2 ~ ;~: ~ ~'7" , ............ :7 ~-:~4~2-. ~ . . . . . ~_~_,. ..... , n ................ ..1,-. ,, :~-- ~ 

Dietary Restriction, N~'P TR 460 

TABLE ~3a 

Summary o1~ the ]Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction StUdy r 

Of Sutyi ~enzy~ Phtha~a~e: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 2A,000 ppm 

2-Year Study (continued) 

Hematopoietic System (continued) 

Lymph node, mandibular (49) (50) (50) 

Ectasia 1 (2%) 2 (4%) 3 (6%) 

Hemorrhage 5 (lO%) 8 (16%) 5 (10%) 

Hyperplasia, lymphoid 6 (12%) 3 (6%) 

Pigmentation 12 (24%) 19 (38%) 16 (32%) 

Lymph node, mesenteric (50) (50) (50) 

Ectasia 1 (2%) 

Hemorrhage 3 (6%) 2 (4%) (4%) 

Hyperplasia, lymphoid 1 (2%) 2 (4%) 

Spleen (50) (50) (50) 

Developmental malformation 1 (2%) 

Fibrosis 1 (2%) 3 (6%) 1 (2%) " 

Hematopoietic cell proliferation 10 (20%) 4 (8%) 14 (28%) 

Hemorrhage 1 (2%) 

Necrosis 1 (2%) 

Pigmentation, hemosiderin 19 (38%) 13 (26%) 29 (58%) 

Thymus (49) (49) (48) 

Hemorrhage 1 (2%) 1 (2%) 

Integumentary System 

Mammary gland (49) (48) (50) 

Ectasia 12 (24%) 5 (10%) 11 (22%) 

Hyperplasia 30 (61%) 9 (19%) 17 (34%) 

Skin (50) (50) (50) 

Aeanthosis 1 (2%) 1 (2%) 

Cyst epithelial inclusion 1 (2%) 1 (2%) 

Hyperkeratosis 3 (6%) 4 (8%) 

Inflammation, chronic 1 (2%) 

Ulcer 2 (4%) 1 -(2%) 2 (4%) 

Musculoskeletal System 

Bone (50) (50) (50) 

Cranium, osteopetrosis 7 (14%) 3 (6%) 6 (12%) 

Femur, osteopetrosis 5 (10%) 4 (8%) .. 3 (6%) 

Rib, osteopetrosis 1 (2%) 1 (2%) 

Skeletal muscle (1) (1) 

Hemorrhage 1 (100%) 1 (lOO%) 

Nervous System 

Brain (50) (50) (50) 

Compression 9 (18%) 1 (2%) 8 (16%) 

Gliosis 1 (2%) 

Hemorrhage 1 (2%) 

Hydrocephalus 2 (4%) 1 (2%) 1 (2%) 

Necrosis 1 (2%) 

161 

i 
162 Dietary Restriction, NTP TR 460 

TABL~ B3a 

SummaE~' of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Respiratory System 

Lung 

Congestion 

Edema 

Hemorrhage 

Infiltration cellular, histiocyte 

Inflammation, subacute 

Alveolar epithelium, hyperplasia 

Nose 

Exudate 

Foreign body 

Fungus 

Mucosa, hyperplasia 

Mucosa, metaplasia, squamous 

Special Senses System 

Eye 

Atrophy 

Cataract 

Hemorrhage 

Retina, degeneration 

Urinary System 

Kidney 

Calculus, microscopic observation only 

Cyst 

Hydronephrosis 

Mineralization 

Nephropathy 

Renal tubule, atrophy 

Renal tubule, cytoplasmic alteration 

Renal tubule, dilatation 

Renal tubule, necrosis 

Renal tubule, pigmentation 

Transitional epithelium, hyperplasia 

Ureter 

Dilatation 

Transitional epithelium, hyperplasia 

Urinary bladder 

Edema 

Transitional epithelium, hyperplasia 

Ad ILibitumFed Control 

. (50) 

21 ,(42%) 

I '(2%) 

2 (4%) 

(50) 

6 (12%) 

5 (10%) 

3 (6%) 

3 (6%) 

3 (6%) 

(2) 

1 (50%) 

I (50%) 

(5O) 

43 (86%) 

34 (68%) 

1 (2%) 

1 (2%) 

1 (2%) 

49 (98%) 

(50) 

i (2%) 

4 (8%) 

Weight-Matched 

Control 

(50) 

I (2%) • 

l (2%) 

17 (34%) 

1 (2%) 

(50) 

3 (6%) 

1 (2%) 

2 (4%) 

3 (6%) 

(2) 

2 (100%) 

2 (100%) 

(5O) 

I (2%) 

2 (4%) 

49 (98%) 

32 (64%) 

48 (96%) 

4 (8%) 

(50) 

24,000 ppm 

(50) 

1 (2%) 

1 (2%) 

19 (38%) 

1 (2%) 

2 (4%) 

(50) 

4 (8%) 

.,. 

I (2%i 

I (2%) 

(3) 

2 (67%) 

1 (33%) 

2 (67%) 

(5O) 

1 (2%) 

2 (4%) 

l (2%) 

35 (70%), 

45 (9o%) 

2 (4%) 

1 (2%). -

1 (2%) 

47 (94%) 

4 (8%) 

(1) 

1 (I00%) 

1 00O%) 

(50) 

10 (20%) 
:7~ -~-~-~.:~ == __~ ,, - , ........................... ~....... 

Dietary Restriction, ~ TR ,~ 1~ 

TABLE B3b 

Summary o~ the Incidence o~ Normeopnastlc Lesions in Female ]Rats in ~he Dietary Restriction Study 

o~ ~uty~ ll~enzy~ Phth~ate: 2-Year .and 32-1V~onth Restricted Feed ~Plrotoco~s a 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

Disposition Summary 

Animals initially in study 60 60 50 50 

15-Month interim evaluation 10 10 

Early deaths 

Moribund 11 10 34 38 

Natural deaths 4 1 6 1 

Survivors 

Terminal sacrifice 35 39 10 11 

Animals examined microscopically 50 50 

15-Month Interim Evaluation 

Alimentary System 

Liver (10) (10) 

Basophilic focus 10 (100%) 3 (30%) 

Clear cell focus 1 (10%) 1 (10%) 

Granuloma 1 (10%) 

Hepatodiaphragmatic nodule 2 (20%) 1 (10%) 

Inflammation, subacute 1 (10%) 

Mixed cell focus 1 (10%) 

Mesentery (1) 

Hemorrhage 1 (100%) 

Fat, necrosis 1 (100%) 

Pancreas (10) (10) 

Atrophy 1 (10%) 1 (10%) 

Stomach, glandular (10) (10) 

Mineralization 1 (10%) 

Cardiovascular System 

Heart (10) (10) 

Cardiomyopathy 2 (20%) 

Endocrine System 

Adrenal cortex (10) (10) 

Accessory adrenal cortical nodule 3 (30%) 3 (30%) 

Angieetasis 1 (10%) 

Pituitary gland (10) (10) 

Pars distalis, angiectasis 1 (10%) 

Pars distalis, cyst 3 (30%) 

Pars distalis, hyperplasia, focal 3 (30%) 

Pars intermedia, cyst 1 (10%) 

Pars intermedia, hemorrhage 1 (10%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
164 Dietary Restriction, NTP TR 460, 

TABLE B3b 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of Bmyl Benzyl Phthalate: 2-Year and 32-Month Restricted Feed Protocols (continued) ..... • ~ , 

 ̧ . . 

• . ' ....... 2-Year Restricted Feed " 32-Month Restricted Feed 

: 0 ppm 

15-Month Interim Evaluation (continued) 

Genital System 

Clitoral gland 

Ectasia 

Inflammalion, chronic 

Uterus 

Hydrometra 

Hyperplasia, cystic • ~ 

Metaplasia, squamous 

Hematopoietic System: 

Bone marrow 

Myclofibrosis 

Eymph node 

Mediastinal, hemorrhage 

Mediastinal, pigmentation 

Lymph node, mandibular 

Hemorrhage 

Pigmentation 

Lymph node, mesenteric 

Hemorrhage 

Spleen 

Hematopoietic cell proliferation 

Pigmentation, hemosiderin 

Thymus 

Cyst 

Integumentary System 

Mammary gland 

Hyperplasia 

Skin 

Ulcer 

Musculoskeletal System 

Bone 

Femur, osteopetrosis 

Respiratory System 

Lung 

Infiltration cellular, histiocyte 

Nose 

Mucosa, hyperplasia 

(1o) 

:~ (2o%) 

(lO) 

2 (20%) 

1 00%) 

1 (10%) 

(IO) 

i (!o%) 

(1) 

1 (1oo%) 

1 (lOO%) 

(10) 

(1oi 

(lO) 

3 (30%i 

9 (90%) 

(1o) 

1 (10%) 

~ 

(10) 

2 (20%) 

(10) 

(10) 

. 

(10) " 

2 (20%) 

(10) 

1 (lO%) 

24,000 ppm 0 ppm 24,000 ppm 

(10) 

1 00%) 

1 (lo%) 

(10) 

1 (10%) 

1 (10%) 

(10) 

2 (20%) 

(1) 

(10) 

3 (30%) 

2 (20%) 

(10) 

1 (10%) 

, (lO) 

1 (10%) 

9 (90%) 

(lO) 

(10) .

(9) ~ 

I (11%) ,.. 

(lO) 

1 (lO%) 

(lo) 

5 (50%) 

(lO) 
~ ......... Tr ......................... ~ . . . . . ~ ? ~ : : Z ~ - ~ : ~ , .~w ....... ~. ~ ....... ,, 

Die~ry Restriction, NTIP TI~ ~6~ ~65

T~L~ ~3b 

Summary o~ the ]Incidence of Nonneopgastic Lesions ~n Female ~ats in the Dietary ]Restriction Study 

o1~ ~uty~ ~enzy~ ]Phtha~ate: 2-3{ear and 32-Month Restricted ]Teed ]Protocols (continued) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

15-Month Inter~m Evaluation (continued) 

Special Senses System 

Eye (1) 

~ 

I Cataract 1 (100%) 

Retina, degeneration 1 (100%) 

Urinary System 

Kidney Cyst (10) (1o) 

i (1o%) 

I~ Mineralization 10 (100%) 8 (80%) 

] Nephropathy 10 (100%) 9 (90%) 

:1 Renal tubule, atrophy 2 (20%) 

Renal tubule, dilatation I (10%) 

i 

• 

Renal tubule, pigmentation 10 (100%) 1o (1oo%) 

Systems Examined With No Lesions Observed 

General Body System 

Nervous System 

2-Year and 32-Month Protocols 

Alimentary System 

Intestine large, colon (50) (49) (49) (50) 

Edema 1 (2%) 

Parasite metazoan 6 (12%) 2 (4%) 5 (10%) 2 (4%) 

Intestine large, rectum (50) (50) (48) (50) 

Parasite metazoan 3 (6%) 3 (6%) 3 (6%) 6 (12%) 

Intestine large, cecum (50) (50) (50) (50) 

Edema 1 (2%) 1 (2%) 

Parasite metazoan I (2%) 

Intestine small, duodenum (50) (50) (49) (50) 

Erosion 1 (2%) 1 (2%) 

Intestine small, jejunum (50) (50) (50) (50) 

Fibrosis 1 (2%) 

Perforation 1 (2%) 

Muscularis, hyperplasia 1 (2%) 

Liver (50) (50) (50) (50) 

Angiectasis 1 (2%) 4 (8%) 1 (2%) 

Basophilic focus 40 (80%) 41 (82%) 32 (64%) 38 (76%) 

Clear cell focus 3 (6%) 2 (4%) 2 (4%) 

Developmental malformation 1 (2 %) 1 (2%) 

Eosinophilic focus 13 (26%) 21 (42%) 8 (16%) 13 (26%) 

Granuloma . 8- (16 % ) 8 (16%) 11 (22%) 5 (10%) 

Hematopoietic cell proliferation 3 (6%) 1 (2%) 

Hemorrhage 1 (2%) 

Hepatodiaphragmatic nodule 3 (6 %) 3 (6%) 2 (4%) 10 (20%) 

Inflammation, subacute 3 (6%) 3 (6%) 1 (2%) 
166 Dietary Restriction, NTP TR 460 

TABt,E B3b 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and 32-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,000 ppm 

2-Year and 32-Month .Protocols (continued) 

Alimentary System (continued) 

Liver (continued) (50) 

Mixed cell focus 5 (10%) 

Thrombosis 

Bile duct, hyperplasia 4 (8%) 

Centrilobular, necrosis 

Hepamcyte, hyperplasia, focal 

Hepatocyte, vacuolization cytoplasmic 

Kupffer cell, pigmentation 1 (2%) 

Lobules, necrosis 6 (12%) 

Mesentery (1) 

Accessory spleen 1 (100%) 

Fat, necrosis 

Pancreas (47) 

Atrophy 27 (57%) 

Acinar cell, hyperplasia, focal 

Aeinus, metaplasia, goblet cell 

Acinus, cytoplasmic alteration 

Salivary glands . (49) 

Duct, cyst 

Stomach ~, forestomach (49) 

Edema 

Ulcer 1 (2%) 

Mucosa, hyperplasia 1 (2%) 

Stomach~ glandular (50) 

Erosion I (2%) 

Ulcer 

Cardiovascular System 

Blood vessel • (49) 

Hypertrophy 

Thrombosis 

Heart 

Cardiomyopathy 

Thrombosis ~, 

(So) 

.15 (3o%) 

-Endocrine System 

Adrenal cortex (50) 

Accessory adrenal cortical nodule 12 (24%) 

Angiectasis . 27 (54 %) 

Degeneration, (atty 11 (22%) 

Hematopoietic cell proliferation 

Hemorrhage 

Hyperplasia; diffuse ......... 

Hyperplasia, focal , 4 (8%) 

Hypertrophy, focal 8 (16%) 

Necrosis 

(50) 

5 (10%) 

1 (2%) 

1 (2%) 

5 (10%) 

2 (4%) 

(50) 

16 (32%) 

1 (2%) 

(50) 

(50) 

(50) 

I (2%) 

1 (2%) 

(50) 

1 (2%) 

1-(2%) 

(50) 

13 (26%) 

(50) 

15 (30%) 

33 (66%) 

7 (14%) 

4 (8%) 

..... 1 (2%) 

5 (10%) 

3 (6%) 

(50) 

11 (22%) 

13 (26%) 

1 (2%) 

6 (12%) 

9 (18%) 

4 (8%) 

(5i

3 (6O%) 

2 (40~) 

(50) • 

25 (50%) 

(~o) 

(48) 

2 (4%) 

4 (8%) 

(48) 

4 (8%) 

(50) 

(50) 

23 (46%) 

1 (2%) 

(5o) 

8 (16%) 

27 (54 %) 

16 (32%) 

6 (!2%) 

7 (14%) 

I (2%) 

•(50) 

l (2%) 

2 (4%) 

11 (22%) 

21 (42%) 

4 (8%) 

(4) 

.I (25%) 

(50) 

22 (44%) 

1 (2%) 

I (2%) 

(50) 

i (2%) 

(50) 

1 (2%) 

1 (2%) 

4 (8%) 

(50) 

., 2 (4%) 

(50) 

(50) 

26 (52%) 

I (2%) , , 

(50) 

17 (34%) 

37 (74%) 

!3 (26%) 

1 (2%) 

4 (8%) 

• , 

3 (6%) 

2 (4%) 
167 

-~z-~--~.__~--~ ..,, , - ~ ~-~_ :::_~=-~_~.~ ~ ~ . . ~ _..: ...... 

D~e~ry'Restriction, ~ T~ 46~ 

TA~L~ ~3b 

Sanmaary o~ the ~nc~dlence o~ Nonneop~ast~c Les~oas ~a ~'ema~e ~ats ~n the Dietary Restr~ct~oa Stadly 

o~ ~uty~ ~e~zy~ ~tha~ate: 2-Year and 32-1V~onth ~estr~cted ~ee~ ]?rotoco~s (continued) 

2-Year Restricted ~'eed 32-Month Restricted ]Need 

.0 ppm 24,000 ppm 0 ppm 24,~ ppm 

2-Year and 32-Month Protocols (continued) 

Endocrine System (continued) 

Adrenal medulla 

Hyperplasia 

(49) 

3 (6%) 

(50) 

1 (2%) 

(49) 

10 (20%) 

(50) 

9 (18%) 

Islets, pancreatic (47) (50) (50) (50) 

Hyperplasia 1 (2%) I (2%) 

Metaplasia, hepatocyte 1 (2%) I (2%) 

Parathyroid gland (49) (46) (48) (49) 

Angiectasis l (2%) 

Cyst 1 (2%) 

Pituitary gland (50) (49) (50) (50) 

Pars distalis, angiectasis 8 (16%) 4 (8%) 8 (16%) 6 (12%) 

Pars distalis, cyst 14 (28%) 10 (20%) 10 (2O%) 11 (22%) 

Pars distalis, hyperplasia, focal 11 (22%) 6 (12%) 9 (18%) 16 (32%) 

Pars intermedia, angiectasis 1 (2%) 1 (2%) 2 (4%) 3 (6%) 

Pars intermedia, cyst 4 (8%) 2 (4%) 3 (6%) 6 (12%) 

Thyroid gland (49) (50) (50) (50) 

Ultimobranchial cyst 1 (2%) 

C-cell, hyperplasia 1 (2%) 5 (10%) 9 (18%) 6 (12%)• 

Follicle, cyst 2 (4%) 4 (8%) 2 (4%) 

General Body System 

None 

Genital System 

Clitoral gland (50) (50) (5o) (49) 

Ectasia 11 (22%) 6 (12%) 15 (3o%) 12 (24%) 

Hyperplasia 1 (2%) 1 (2%) 4 (8%) 1 (2%) 

Inflammation, chronic 3 (6%) 4 (8%) 1 (2%) 

Inflammation, suppurative '5 (10%) 3 (6%) 7 (14%) 2 (4%) 

Ovary (50) (50) (50) (50) 

Cyst 3 (6%) 6 (12%) 6 (12%) 6 (12%) 

Uterus (50) (50) (50) (5O) 

Hydrometra ~ 3 (6%) 7 (14%) 2 (4%) 2 (4%) 

Hyperplasia, cystic 7 (14%) 16 (32%) 7 (14%) 7 (14%) 

Hematopoietic System 

Bone marrow (50) (50) (50) • (50) 

Hypcrccllularity " 1 (2%) 2 (4%) 1 (2%) 

Hypcrplasia, rcticulum cell 1 (2%) 

Myelofibrosis 2 (4%) 3 (6%) 5 (10%). 

Necrosis 1 (2%) 
168 Dietary Restriction, NTP TR ~d} 

TA~t~B3b • . 

Summary of the Incidence of Nonneoplastic Lesions in,Female Ratsin the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and 32-Month Restricted Feed Protocols (continuEd) 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm ; 0 ppm 24,000 ppm 

. 

2-Year and 3~=~Month Protocols (continued) 

Hematopoietic System (continued) 

Lymph node (10) (8) (21) (18) 

Iliac, ectasia 2 (10%) 

Iliac, hyperplasia, lymphoid 

Inguinal, hyperplasia, lymphoid 

Mediastinal, hemorrhage 

,,, 

1 (1o%) 

.. 

1 (13%) 

2 (10%) 

1 (5%) 

1 (5%) I (6%) 

Mediastinal, hyperplasia, lymphoid 1 (13%) 

Mediasdmi, pigmentation 

Pancreatiq, pigmentation 

... .... .. 3 (30%) 

1 (1o%) 3 (38%) 

7 (33%) 

2 (10%) 

3 07 %) 

4 (22%) 

Renal, hemorrhage 

Renal, pigmen.tatipn 

Lymph node, mandibular 

l~ctaSia 

1 (lo%) • 

(49), • 

3 (6%) 

1 (13%) 

2 (25%) 

(50) 

2 (4%) 

2 (10%) 

.(5o) 

3 (6o%) 

, 

4 (22%) 

(50) 

7 (14%) 

Hemorrhage .. 

Hyperplasia, lymphoid 

Pigmentation 

Lymph node; mesenteric " 

Hemorrhage 

Hyperplasia, lymphoid 

Spleen 

Fibrosis • ':, 

Hematopoietic cell proliferation 

Hemorrhage 

Necrosis 

' 5 (1o%) ,: 

.15 (31%) 

(50) 

• 2 (4%) 

t (2%) 

(5o) 

3 (6%) 

' " 13 (26%) 

2 (4%) ,. 

7 (14%) 

10 (20%) 

(50) 

2 (4%) 

1 (2%) 

(50) 

2 (4%) 

8 (16%) 

1 (2%) 

,, 

6 (12%) 

,,1 (2%) 

19 (38%) 

(50) 

2 (4%) 

1 (2%) 

(50) 

6 (12%) 

13 (26%) 

7 (14%) 

6 (12%) 

17 (34%) 

(50) 

2 (4%) 

! (2%) 

(50) 

7 (14%) 

8 (16%) 

1 (2%) 

Pigmentation, hemosideriri ....... 

Thymus 

Atrophy 

'Hemorrhage . 

' 2i (42%) 

(50) 

18 (36%) 

(50) 

1 (2%) 

" 18 (36%) 

(49) 

1 (2%) 

2~ (48 %)" 

(46) • 

Integumentary System 

Manunary gland' 

Ectasia 

(50) ............ 

16 (32%) 

(49) ............ 

5 (10%) 

(49) ......... 

22 (45%) 

(50) 

• 12 (24%) 

Galactocele 

. Hyperplasia • . 

Skin 

Acamh0sis 

Cyst epithelial inclusion 

Edema : • 

Hyperkeratosis . 

Inflammation, chronic 

._Ulcer 

-.. 

' 

: ,. 

; 1 (2%) 

.18 (36%) 

(50) 

3 (6%) 

/i1 (2%) 

3 (6%) 

t. 

.. 

~ " -. 

18 • 

(50) 

~ 5 

3 

(37%i 

(10%) 

(6%) 

2 (4%) 

20 (41%) 

(50) 

2 (4%) 

2 (4%) 

• 1 (2%) 

32 .(64%) 

(50) 

6 (12%) 

1 (2%) . 

2 (4%) 

2 (4%) 

1 (2%) 

4 (8%) . . 

:. 

Musculoskeletal System . 

(50) (50) . Bone .,':(5oi ~ (50) 

. 6 (12%) 9 (18%) Cranium, osteopetrosis • 12 (24%) .... "" 2 (4% i 

• 7 (14%) 8 (16%) Femur, osteopetrosis " ~ ..... 7(14%) " ,4 (8%) 

1 (2%) .~Rib, osteopetrosis : 1 (2%) . 

, 
: ........ ~_ ,, . . . . . . . ~i¢,im r- ~ ...... ~ i ~ ..... z . . . . . .--, ~ ~ . . . . . . . . . . . . ~ 2 _ 

Dietary Restriction, NTP T~ 460 

TABLE S3b 

Summary of the ]Incidence of Nonneopimtic Lesions ira ~ema~e lI~ats in the D~et~'y Restriction Study 

of ~uty~ ~enzy~ Phth~date: 2-Year and 32-Month ~estr~cted ~'eed ]~-ot~coL~ (continu~) = ' r 

2-Year Restricted Feed 32-Month Restricted Feed 

0 ppm 24,000 ppm 0 ppm 24,0110 ppm 

2-Fear and 32-Month Protocols (continued) 

Nervous System 

Brain (50) (50) (50) (49) 

Compression 5 (1o%) 5 (10%) 14 (28%) 7 (14%) 

Hemorrhage I (2%) I (2%) • 2 (4%) 

Hydrocephalus 1 (2%) 3 (6%) 3 (6%) 1 (2%) 

Inflammation, chronic I (2%) 

Necrosis I (2%) : 

Respiratory System 

Lung (50) (50) (50) (50) 

Congestion 2 (4%) 1 (2%) 

Edema i (2%) 1 (2%) 

Hemorrhage 3 (6%) 1 (2%) 

Inf'fltration cellular, histiocyte 18 (36%) 18 (36%) 14 (28%) 21 (42%) 

Inflammation, subacute 1 (2%) 1 (2%) 1 (2%) 

Alveolar epithelium, hyperplasia 5 (10%) I (2.%) 4 (8%) 1 (2%) 

Nose (50) (50) (50) (50) 

Exudate 3 (6%) 6 (12%) 6 (12%) 

Foreign body 1 (2%) 1 (2%) I (2%) 

Fungus 1 (2%) 2 (4%) I (2%) 

Mucosa, hyperplasia 1 (2%) 3 (6%) 3 (6%) 3 (6%) 

Mucosa, metaplasia, squamous 3 (6%) l (2%) i 

Special Senses System 

Eye (3) (2) (1) (2) 

Atrophy 1 (lOO%) 

Cataract 2 (67%) 2 (100%). ~ , . . . . . 2 (100%)

Retina, degeneration 2 (67%) 2 (100%) 2 (100%) 

Urinary System 

Kidney (50). (50) (50) (50) 

Calculus, microscopic observation only 8 (16%) 1 (2%) ,9 (18%) 

Cyst 1 (2%) 

Hydronephrosis ' 1 (2%) 

Inflammation, suppurative 2 (4%) 

Mineraalization .46 (92%) 34 (68%) 45 (90%) 31 (62%) 

Nephropathy ' ,40 (80%) 47 (94%) 40 (80%) 49 (98%)' 

Papilla, necrosis 3 (6%) 

Renal tubule, atrophy 3 (6%) 4 (8%) 

Renal tubule, dilatation (lO%) ....... t2. (24%). 

Renal tubule, hyperplasia t (2%) 1 (2%) 

Renal tubule, necrosis 1 (2%) 

Renal tubule, pigmentation 48 (96%) 50 (100%) 48 (96%) 48 (96%) 

Transitional epithelium, hyperplasia 1 (2%) 20 (40%) 2 (4%) 29 (58%) 

Urinary bladder • (50) (50) (49) (50) 

Calculus, microscopic observation only I (2%) 

Transitional epithelium, hyperplasia 14 (28%) 16 (32%) 
170 

Dietary Restriction, NTP TR 460 
:: ~'~:~-~'~--i'~'W ~_~_ ~, .............. , ...... = ~ ....... ~, --.~,_ ~-.~'~-=~- ---~.~--~ ...... : ............ ~.~_ 

T~ C1a 

T~ Clb 

T~ C2a 

T~L~ C2b 

T~ C3~ 

TABLE C3b 

~71 

APPENII~IX C 

~UMI~JIA~Y OF ILE~II©N~ IN MAILE ~AT~ 

IN TI~i]E DIIETA~Y XE~T~ICTI©N ~TUI~Y 

OF ~-IlIUTYILI-IIYID~Oq!UIHOHE 

S~mm-y of the Incidence of Neoplasms in Male Rats 

nu the Dietary Restriction Study of t-~utylhydroqu~none: 

Ad ~b~m ~eed~ng and Weight-Matched Control ~ot~o~ ................... 

Summary of the ~nddence o~ Neopl~ms ~n ~ale ~ts 

m the Dietary ~tr~cfion Stu@ of t-Butylhydroqugnone: 

~r~cted ~eed P~ot~oa ........................................... 

S~at~ca~ Analysis of ~a~ Neopl~ms ~n Male ~ 

m the D~e~y ~tr~cfion Study of t-Butylhydroqu~none: 

Ad LD~m ~eed~ng and Wright-Matched Controls Prot~ois ................. 

Stafisfica~ Anaays~s of PHma~ Neopl~ms in Male ~ats 

m the D~eta~y ~tr~cfion Study of t-Buty~hydroqu~none: 

~tr~cted ~eed ~rotoco~ ................................. .......... 

.. 

172 

177 

111 

119 

i 

Su~a~ o~ the ~ndfience o~ Nonneopl~fic L~o~ ~n Male ~a~ 

m the ~eta~ ~tHcfion Study o~ t-Buty~hydroqu~none: 

Ad ~b~m ~eed~ng and Wdght-~atched Controls P~ot~o~ ................... 

Sum~a~ of the ~ncidence o~ Nonneopl~fic Lesions ~n ~a~e ~a~ 

m the ~eta~y ~t~ct~on Study o~ t-Butylhyd~oqu~none: 

~t~[cte~ ~ee~ P~ot~o~ ........................................... 

~9~ 

20~ 

i 
172 Dietary Restriction, NTP TR 460 

TA~t,E Cla 

Sunnnary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydro~ uinone:" -

Ad .Libitum Feeding and Weight-Matched Controls Protocols a --

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

Disp~ositionSummary 

Animals initially in study 70 70 70 

3-Month interim evaluation 10 10 10 

Early deaths 

Moribund 48 41 42 

Natural deaths 4 " 7 4 

Survivors 

Terminal sacrifice . 8 12 14 

Animals examined, microscopically 70 70 70 

/ 

Sy.ste~ns Examined At 3 Months With No Neoplasms Observed 

Alimentary System . 

Cardiovascular System 

Endocrine System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System ~. 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

30-Month Study 

Alimentary System 

Intestine large, colon (58) (60) (6O) 

Intestine large, rectum " (59) (58) (59) 

Intestine :large,. cecum .... (60) .... ........ (60) . . . . . (60) 

Intestine small, duodenum (60) (60) (60) 

Intestine small, jejunum (60) (59) (60) 

Intestine small, ileum (60) (59) (59) 

Liver (60) . (6o) ..................... (60) ~. 

Hemangiosar~orna 1 (2%) 

Hepatocellular carcinoma 2 (s%) 

Hepatocdlular adenorna 4 (7%) I (2%) 3 (5%) 

Histiocytic sarcoma l (2%) 

Mesentery (20) (15) • (16) 

Histiocytic sarcoma I (5%) 

.Schwannoma malignant I (5%)~ 

Oral mucosa (I)" 

Squamous cell carcinoma 1 (I00%) 

Pancreas (6O) 

Acinus, adenoma 

Mixed tumor benign 

Salivary glands 

', 

' : 

• 3 

(60) 

(5%) 

1 (2%) 

(60) (60) 
173 

~ ~ ~-~= ,~-~v~_,~,-- ~ 

Dietary ]Restriction, NTP TR 460 

TA~L~ Cla 

Summary o~ the Incidence o~ NeoPlaSmS ~n IV~a~e ~ta~. in the Dietary lRestricfion Study o~ t-~uty~hydroqu~none: 

Ad Libitum ]Feeding and Weight-Matched Control Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

30-Month Study (continued) 

Alimentary System (continued) 

Stomach, forcstomach 

Squamous cell papilloma 

Stomach, glandular 

Tongue 

l Sqnamous cell carcinoma 

Squamous cell papilloma 

i 

Cardiovascular System 

Heart 

Schwannoma malignant 

(6O) 

(6O) 

(1) 

1 (100%) 

(6o) 

1 (2%) 

(6O) 

2 (3%) 

(6O) 

(2) 

1 (50%) 

(6o) 

(59) 

1 (2%) 

(60) 

(4) 

2 (50%) 

(6o) 

Endocrine System 

Adrenal cortex (6O) (6O) (6O) 

Adenoma 

Adrenal medulla 

1 

(6O) 

(2%) 1 (2%) 

(6O) 

1 

(6O) 

(2%) 

Pheochromocytoma malignant 1 (2%) 1 (2%) 3 (5%) 

Pheochromocytoma complex 1 (2%) 

Pheochromocytoma benign 13 (22%) 11 (18%) 11 (18%) 

Pheochromocytoma benign, multiple 

Islets, pancreatic 

1 

(6O) 

(2%) 1 (2%) 

(6O) 

1 

(6O) 

(2%) 

Adenoma 5 (8%) 2 (3%) 3 (5%) 

Carcinoma 

Pituitary gland 

1 

(6O) 

(2%) 

(59) (6O) 

Pars distalis, adenoma 

Pars distalis, carcinoma 

Thyroid gland 

C-cell, adenoma 

• !9 

(6O) 

5 

(32%) 

(8%) 

19 (32%) 

1 (2%) 

(6O) 

5 (8%) 

6 

1 

(6O) 

4 

(10%) 

(2%) 

(7%) 

C-cell, carcinoma 1 (2%) 2 (3%) 

Follicular cell, carcinoma 3 (5%) 

General Body System 

Peritoneum (I) (3) 

Genital System 

Epididymis (6o) (60) (6o) 

Preputial gland (6o) (6o) (6O) 

Adanoma 5 (8%) 3 (5%) 

Carcinoma 2 (3%) 5 (8%) 

Prostate (6o) • (6o) (6o) 

Adenoma 2 (3%) 3 (5%) 1 (2%) 

Seminal vesicle (6o) (6o) (6o) 

Testes (60) • (6o) (6o) 

Bilateral, interstitial cell, adenoma 42 (70%) 53 (88%) 50 (83%) 

Interstitial cell, adenoma 13 (22%) 5 (8%) 9 (15%) 

i 
174. Dietary Restriction, NTP TR 460 

TABLE Cla 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Ad Libitum and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

30-Month ,Study (continued) 

Hematopolefic System 

Bone marrow (6O) (6O) (60) 

Lymph node (33) (33) (35) 

Lymph node, mandibular (60) (59) (60) 

Carcinoma, metastatic, Zymbal's gland 1 (2%) 

Lymph node, mesenteric (6O) (58) (60) 

Hemangioma " I (2%) 

Spleen (60) (6O) (60) 

Fibroma 2 (3%) 1 (2%) 

Histiocytic sarcoma 1 (2%) 

Hemangiosarcoma 1 (2%) 

Sarcoma I (2%) 1 (2%) 

Thymus (58) (55) (56) 

Thymoma malignant I (2%) 

Integumentary System 

Mammary gland. (57) (58) (58) 

Adenoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

I (2%) 

9 (16%) 

1 (2%) 

(60) 

I (2%) 

6 (10%) 

(60) 

6 (10%) 

1 (2%) 

(60) 

Basal cell carcinoma 1 (2%) 

Hemangioma 

Keratoacanthoma 3 (5%) 

,~ (2%) 

2 (3%) 4 (7%) 

Keratoacanthoma, multiple 

Squamous cell carcinoma, metastatic, tongue 

Squamous cell papilloma 

Trichoepithelioma 

t (2%) 

2 (3%) 

2 0%) 

I (2%) 

1 (2%) 

1 (2%). 

2 (3%) 

1 (2%) 

Sebaceous gland, adenoma 

Subcutaneous tissue, fibroma 

1 (2%) 

3 (5%) 7 (12%) 6 (10%) 

Subcutaneous tissue, fibroma, multiple 

Subcutaneous tissue, fibrosarcoma 

I (2%) 

1 (2%) 3 (5%) 1 (2%) 

Subcutaneous tissue, hemangioma '.... .... ~ (2%) • . 

Subcutaneous tissue, hemangiosarcoma I (2%) 

Subcutaneous tissue, schwannoma benign 

Subcutaneous tissue, schwannoma malignant 

~ (2%) 1 (2%) 

Musculoskeletal System 

None 

Nervous System 

Brain (60) (60) (60) 

Astrocytoma malignant 

Carcinoma, metastatic, pituitary gland 

1 (2%) 1 (2%) 

Oligodendroglioma malignant 

Spinal cord 

1 (2%) 

(2) (1) 
~ ~ ~ ............ i~ ~ ~ ~ ¢ ............... " .......... _ _ ~ : ~ 1 - - ......... ~ 

D~etmT ~est~ic~on, I~T~ T~ ~ ~175 

T~L~ Cga 

S~mn~'y o~ the Incidence o~ Iqeo#gsms in M~e RaN in the Die~a~ Restriction Study o~ g-~ty~hydroqa~one: 

Ad Libigum ~nd Weight-Matched Controls ~Prot~o~s (continued) 

Ad Lib'.~m- Weight-Matched 

Fed Control Control 5,0~ ppm 

~30-Month Study (continued) 

Respiratory System 

Lung (60) (60) (60) 

Aiveolar/bronchiolar adenoma 3 (5%) 2 (3%) I (2%) 

Alveolar/bronchiolar carcinoma 3 (5%) 1 (2%) 

Histiocytic sarcoma 1 (2%) 

Nose (60) (6O) (6O) 

Squamous cell papilloma 1 (2%) 

Special Senses System 

Eye (4) (4) (2) 

Schwannoma malignant t (25%) 

Zymbal's gland (2) (1) (4) 

Adenoma 1 (25%) 

Carcinoma 2 (100%) I (100%) 3 (75%) 

Urinary System 

Kidney (60) (6O) (60) 

Lipoma 1 (2%) 

Renal tubule, adenoma 2 (3%) 1 (2%) 

Transitional epithelium, carcinoma 1 (2%) 

Urinary bladder (60) (60) (60) 

Papilloma 1 (2%) 

Systemic Lesions 

Multiple organs b (6o) (60) (60)

Histiocytic sarcoma 1 (2%) 

Leukemia mononuclear 39 (65%) 45 (75%) 32 (53%) 

Lymphoma malignant 1 (2%) 

Mesothelioma malignant 1 (2%) 3 (5%) 
176 Dietary Restriction, NTP TR 460 

TABLE Cla 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Ad Libitum and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

Neoplasm Summary 

Total animals with primary neoplasms ¢ 

28-Month study 60 60 60 

Total primary neoplasms 

28-Month study 207 186 179 

Total animals with benign neoplasms 

28-Month study 60 59 59 

Total benign neoplasms 

28-Month study 148 126 120 

Total animals with malignant neoplasms 

28-Month study 51 • 51 47 

Total malignant neoplasms 

28-Month study 59 60 59 

Total animals with metastatic neoplasms 

28-Month shady 2 2 

Total metastatic neoplasms 

28-Month study 2 2 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

c Primary neoplasms: all neoplasms except metastatic neoplasms 
177 

=~ :Z2~-~2~'~ : Z~ - - - :; ,__ ~ ~_~,,_:~::__ 

Dietary Restriction, NTP TR 460 

TABLE Clb 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of t-ll$utynhydroquhmne: 

Restricted Feed ProtocoP 

0 ppm 5,000 ppm 

Disposition Summary 

Animals initially in s 70 70 

3-Month interim evaluation 10 10 

Early deaths 

Moribund 43 33 

Natural deaths 7 5 

Survivors 

Terminal sacrifice 10 22 

Animals examined microscopically 70 70 

Systems Examined At 3 Months With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

Endocrine System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musculoskidetal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

30-Month Study 

Alimentary System 

Intestine large, colon (60) (60) 

Intestine large, cecum (60) (60) 

Intestine small, duodenum (60) (60) 

Intestine small, jejunum (60) (60) 

Leiomyosarcoma 1 (2%) 

Intestine small, ileum (60) (59) 

Liver (60) (60) 

Carcinoma, metastatic, thyroid gland 1 (2%) 

Mesentery (8) (5) 

Hemangiosarcoma 1 (20%) 

Oral mucosa (2) 

Carcinoma 1 (50%) 

Squamous cell papilloma 1 (50%) 

Pancreas (58) (60) 

Acinus, leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Salivary glands (60) (60) 

Stomach, forestomach (60) (60) 

Leiomyosarcoma 1 (2%) 

Squamous cell papilloma 3 (5%) 
178 Dietary Restriction, NTP TR 460 

TABLE ClLb 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Restricted Feed Protocol (continued) 

30-Month Study (continued) 

Alimentary System (continued) 

Stomach, glandular 

Tongue 

Squamous cell carcinoma 

Squamous cell papilloma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex • 

Carcinoma, metastatic, thyroid gland 

Osteosarcoma, metastatic, bone 

Adrenal medulla 

Pbeochromocytoma malignant 

Pheochromocytoma complex 

Pheochromocytoma benign 

Pheochromocytoma benign, multiple 

Islets, pancreatic 

Adenoma 

Adenoma, multiple 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

C-ceil, adenoma, multiple 

C-cell, carcinoma 

Follicular cell, carcinoma 

General Body System 

Peritoneum 

Genital System 

Preputial gland 

Adenoma 

Carcinoma 

Prostate 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

0 ppm 

(60) 

0) 

1 (100%) 

(60) 

(60) 

(6o) 

~ (2%) 

(59) 

2 (3%) 

1 (2%) 

7 (12%) 

5 (8%) 

(58) 

2 (3%) 

(57) 

13 (23%) 

(60) 

7 (12%) 

(3%) 

(3) 

(60) 

1 (2%) 

1 (2%) 

(6O) /

(60) " 

(60) 

52 (87%) 

7 (12%) 

5,000 ppm 

(60) 

(2) 

l (50%) 

(60) 

(60) . 

(60) 

1 (2%) 

(60) 

2 (3%) 

11 (18%) 

1 (2%) 

(60). 

5 (8%) 

1 (2%) 

(56) 

~6 (29%) 

(59) 

5 (8%) 

i (2%) 

5 (8%) 

(3) 

(59) 

1 (2%) 

2 (3%) 

(60) 

(60) 

(60) 

56 (93%) 

4 (7%) 
179 

~ ~ - - ~ ~ ...... , ~ ......... ...... _~ ~ ............ ~:~ ~ ~,~ 

Dietary Restriction, NTP T~ ~ 

TABLE Clb 

Su~nn~ary of the Incidence of Neoplasms in Male mats ~n the Dietary SestHcfion Study of t-Buty~hydroqu[none: 

l~estHcted l~eed ]th'ot~coi (continued) 

0 ppm 5,000 ppm 

. 

3O-Monfk Study(continued) 

Hematopoietic System 

Bone marrow (6o) (6o) 

Lymph node (36) (23) 

Deep cervical, carcinoma, metastatic, 

thyroid gland I (3%) 1 (4%) 

Lymph node, mandibular (60) (60) 

Carcinoma, metastatic, thyroid gland I (2%) 

Lymph node, mesenteric (60) (60) 

Spleen (60) (60). 

Thymus (54) (58) 

Integumentary System 

Mammary gland (58) (58) 

Carcinoma 1 (2%) I (2%) 

Fibroadenoma 4 (7%) 6 (10%) 

Skin (60) (60) 

Basal cell adenoma I (2%) 

Basal cell carcinoma 1 (2%) 

Keratoaeanthoma 1 (2%) I (2%) 

Sebaceous gland, adenoma 1 (2%) 

Subcutaneous tissue, fibroma 6 (10%) 3 (5%) 

Subcutaneous tissue, fibrosar~oma I (2%) 

Subcutaneous tissue, schwannoma malignam I (2%) 

Musculoskeletal System 

Bone (60) (60) 

Ost~osarcoma 2 (3%) 

Nervous System 

Brain (60) (60) 

Astrocytoma malignant l (2%) 

Spinal cord (2) 

Respiratory System 

Lung (60) (6(~ 

Alveolar/bronchiolar adenoma I (2%) 1 (2%) 

Alveolar/bronchiolar adenoma, multiple 1 (2%) 

Alveolar/bronchiolar carcinoma 1 (2%) 

Carcinoma, metastatic, thyroid gland I (2%) 1 (2%) 

Fibrosarcoma, metastatic, skin 1 (2%) 

Leiomyosarcoma, metastatic, stomach, 

for~stomach I (2%) 

Osteosarcoma, metastatic, bone I (2%) 

Nose (60) (~) 

Squamous cell carcinoma I (2%) 

Vomeronasal organ, adenoma I (2%) 
180 

c 

Dietary RestriCtion, NTP TR 460 

. 

TABLE Clb 

Summary of the Incidence Of Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydr0quinone: 

Restricted Feed Protocol (continued) 

0 ppm 5,000 ppm • 

30-Month Study(continued) 

Special Senses System 

Zymbal's gland (1) 

Carcinoma 1 (~00%) 

Urinary System 

Kidney (60) (60) 

Renal tubule, adenoma i (2%) 

Transitional epithelium, carcinoma 1 (2%) 

Urinary bladder (60) (60) 

Papilloma 1 (2%) 

Systemic Lesions 

Multiple organs b (60) (60) 

Leukemia mononuclear 46 (77%) 39 (65%) 

Mesothelioma malignant 3. (5%) ~ (7%) 

Neoplasm Summary 

Total animals with primary neoplasms e 

2-Year study 59 60 

Total primary neoplasms 

2-Year stud), 166 187 

Total animals with benign neoplasms ,60 2-Year study 59 

Total benign neoplasms 

2-Year study 107 122 

Total animals with malignant neoplasms 

2-Year study 52 49 

Total malignant neoplasms 

2-Year study 59 65 

Total animals with metastatic neoplasms 

2-Year study 1 4 

Total metastatic neoplasms 

/ 

7 2-Year study 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically " 

Primary neoplasms: all neoplasms except metastatic neoplasms 
:: ::=. L~ ~-~:-~.~5~7 ~ ...... "-- .~,~--r:; ...... , ...... ......... ~-_~ . . . . . . . . . . . . . . . :~'~'~==~'~ ......... ~ ":-" - ...... ; ,,., 

Dietary ll~estriction, NT]? T~ 46~ 381 

TABLE C2a 

StafisficagAnalysisof Prhnary Neoplasms in Male Rats in the Dietary ]Restriction Study o~ t-SutY~hydroquinone: 

Ad Libitum ~eeding and Weight-Matched Controls ]Protocols 

5,000 ppm x 5,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Adrenal Medulla: Benign Pheochromocytoma 

Overall rate a 14/60 (23%) 12160 (20%) 12160 (20%) 12160 (20%) 

Adjusted rate b 59.8 % 49.8 % 45.3 % 49.8 % 

Terminal ~ate ¢ 2/8 (25%) 5/14 (36%) 2/12 (17%) 5/14 (36%) 

First incidence (days) 667 487 617 487 

Life table test d P=0.193N P=0.481 

Logistic regression test d P=0,351N P=0.482 

Fisher exact test d P=0.412N P=0.590N 

Adrenal Medulla: Malignant PheochromOcytoma 

Overall rate 1/60 (2%) 3/60 (5%) 1/60 (2%) 3/60 (5%) 

Adjusted rate 3.1% 21.4 % 8.3 % 21.4% 

Terminal rate 0/8 (0%) 3/14 (21%) 1/12 (8%) 3/14 (21%) 

First incidence (days) 710 857 (T) 857 (T) 857 (T) 

Life table test P=0.465 P=0.356 

Logistic regressibn test P=0.394 P=0.356 

Fisher exact test P=0.309 P=0.309 

Adrenal Medulla: Benign, Complex, or Malignant Pheochromocytoma 

Overall rate 14/60 (23%) 13/60 (22%) 13/60 (22%) 13/60 (22%) 

Adjusted*rate. ~9.8% 55.4% 50.8% 55.4% 

Terminal rate 2/8 (25%) 6/14 (43.%) 3]12 (25%) 6/14 (43%) 

First incidence (d~ys) 667 487 617 487 

Life table test P=0.239N P=0.494 

Logistic r~gression test P=0.431N P=0.455 

Fisher exact test P=0.500N P=0.588N 

Liver: Hepatocellular Adenbma 

Overall rate 4/60 (7%) 3/60 (5%) 1/60 (2%) 3/60 (5%) 

Adjusted rate 25.1% • 12.7% 2.2% 12.7% 

Terminal rate 1/8 (13%) 1/14 (7%) 0/12 (0%) 1/14 (7%) 

First incidence (days) 642 708 673 708 

Life table test P=0.370N P=0.285 

Logistic regression test P=0.463N P=0.297 

Fisher exact test P=0.500N P=0.309 

Liver: I-Iepatocellular Adenoma or Carcinoma  ̧ ., 

Overall rate 4/60 (7%) 5/60 (8%) 1/60 (2%) 5/60 (8%) 

Adjusted rate 25.1% 21.0% 2.2% 21.0% 

Terminal rate 1/8 (13%) 1/14, (7%) 0/12 (0%) 1/14 (7%) 

First incidence (days) 642 708 673 708 

Life table test P=0.611N P=0.081 

Logistic regression test P=0.548 P=0.090 

Fisher exact test P=0.500 P=0.103 

i 

I = 

i 
182 Dietary Restriction, NTP TR 460 ' 

TABt,E C2a 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of t-Butyihydroquinone: 

Ad l_dbitum Feeding and Weight-Matched Controls Protocols (continued) 

5,000 ppm x 5,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

• Lung: Alveolar/bronchiolar Adenoma 

Overall rate 3/60 (5 %) 1/60 (2 %) 2/60 (3%) 1/60 (2%) 

Adjusted rate 10.2 % 4.3 % 

Terminal rate 0/8 (0%) 0/14 (0%) 

First incidence (days) 618 773 

Life table test P=0.276N 

Logistic regression test P=0.306N 

Fisher exact test P=0.309N 

Lung: Alveolar/bronchi01ar Carcinoma 

Overall rate 0/60 (O%) 1/60 (2%) 3/60 (5%) 1/60 (2%) 

Adjusted rate 20.1% 7.1% 

Termimd rate 2/12 (17%) 1/14 (7%) 

First incidence (days) 814 857 (T) 

Life table test P=0.301N 

Logistic regression test • P--0.373N 

Fisher exact test P=0.309N 

Lung: Alve01ar/bronchiolar Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

3/6O (5%) 

I0.2% 

2/6O (3%) 

11.,2% 

5/60 (8%) 

30,1% 

2/60 (3%) 

11,2% 

Terminal rate • " 0/8 (0%) 1/14 (7%) 3/12 (25%) 1/14 (7%) 

First incidence (days) 618 773 722 773 

Life table test P=0.418N P=0.226N 

Logistic regression test . P=0.488N P=0.297N 

Fisher exact test P=0.500N P=0.219N 

Mammary Gland: Fibroadenoma 

Overall rate 10/60 (17%) 7/60 (12%) 6/60 (10%) 7/60 (12%) 

Adjusted rate 72.0% 4O.2% 37.9% 40.2% 

Terminal rote , 5/8 (63%) 5/14 (36%) 3/12 (25%) 5/14 (36%) 

First incidence (days) 381 7O8 810 708 

Life table test P=0.059N P=0.527 

Logistic :regression test P=0.107N P=0.330 

Fisher exact test • P=0.301N P=0.500 . 

Mammary Gland: Fibroadenoma or Adenoma .. 

Overall rate 11/60 (18%) 7/60 (12%) 7/60 (12%) 7/60 (12%) 

Adjusted rate 72.6% 40.2% 40.8% 40.2% 

Terminal rate 518 (63%) 5/14 (36%) 3/12 (25%) 5/14 (36%) 

First incidence (days) 381 708 810 708 

Life table test P=0.039N ,P--0.609N 

Logistic regression test ~ P=0.076N P=0.439 

Fisher exact test P=0.222N P--0.611N 
:~ ~-'~~~ :~.~.L~~~iz=..~ ~ - .............................. - --~-~,- ~. : ........... ~..~ ............. 

D~e~ry l~est~cfion, ~ T~ ~ ~83 

TABLE C2a 

StatLsfic.a~ Analysis o~ ]~rhnary Neoplasmsin Male Rats in the Dietary Restriction Study o~ t-~uty~hydroqu~none: 

Ad Libit~ra ~eed~ng and We~ght-IV~atched Controls Protocols (continued) 

5,000 ppm x 5,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Pancrem: Adenoma 

Overall rate : 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pancreatic ~slets: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pancreatic ~slets: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Adenoma 

• 

3/60 (5 %) 

37.5 % 

3/8 (38%) 

857 (T) 

5/60 (8 %) 

32.5% 

1/8 (13%) 

667 

6160 (10%) 

" 42.1% 

2/8 (25%) 

667 

19160 (32%) 

63.8% 

2/8 (25 %) 

528 

Adenoma or Carcinoma 

19160 (32%) 

63.8 % 

2/8 (25%) 

528 

0/60 (0 %) 

0.0% 

0/14 (0%) 

-

P=0.038N 

P=0.038N 

P=0.122N 

3/60 (5 %) 

15.9% 

1/14 (7%) 

708 

P=O.208N 

Pf0.287N 

Pffi0.359N 

3/60 (5%) 

15.9% 

1/14 (7%) 

708 

P=O. 112N 

P =0.170N 

P=0.245N 

6/60 (10%) 

30.2% 

3/14 (21%) 

668 

P=0.002N 

P-~0.002N 

P =0.003N 

7/60 (12%) 

31.6% 

3/14 (21%) 

627 

P=O.004N 

P= O.006N 

P =0.007N 

0/6O (0%) 

2/60 (3%) 

10.7% 

1/12 (8%) 

715 

.. 

2160 (3%) 

10.7% 

1/12 (8%) 

715 

19/59 (32 %) 

72.4% 

7/12 (58%) 

638 

20/59 (34%) 

74.2% 

7/12 (58%) 

638 

0/6O (0%) 

3/60 (5%) 

15.9% 

1/14 (7%) 

7O8 

P=0.492 

P=0.432 

P=0.560 

3/60 (5%) 

15.9% 

1/14 (7%) 

708 _ 

P=0.492 

P=0.432 

P=0.500 

6/60 (1o%) 

30.2% 

3/14 (21%) 

668 

P=0.008N 

P=0.007N 

P=0.003N 

7/60 (12%) 

31.6% 

3/14 (21%) 

627 

P=0.011N 

P=0.008N 

P=0.003N 
-- 

-- 

184 Dietary Restriction, NTP TR 460 - -. [ 

- ~ 

TA~t,E C2a ' 

Statistical Analysis of Primary Neoplasms in Male Rats in the DietaryRestriction Study Of t-Butylhydroquinone: 

Ad la!bitumFeeding and Weight-Matched Controls Protocols (continued) • 

5,000 ppm x 5,000 ppm x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Preputial Gland: Adenoma ': 

Overall rate 5/60 (8 %) 3/60 (5 %) 0/6O (0%) 3/6O (5%) 

Adjusted rate 3 I. 1% 18.4 % 0.0% 18.4% 

Terminal rate 2/8 (25%) 2/14 (14%i 0/12 (0%) 2/14 (14%) 

First incidence (days)• 528 786 786 

Life table test P=0.205N P=0.121 

Logistic regression test P=0.312N P=0.084. 

Fisher exact test P=0.359N P=0.122 

~. . . Preputial Gland: Carcinoma 

Overall rate 2/60(3%) 5/6O (8%) 0/60 (0%) 5/6O (8%) 

Adjusted rate 3:6% 18.4% 0.0% 18.4% 

Terminal rate 0/8 (0%) 1/14 (7%) 0/12 (0%) 1/14 (7%) 

First incidence (days) 381 520 520 

Life table test , P=0.287 P=0.027 

Logistic regression test P=0.200 p=o.o4o • 

P=0.219 P=0.029 ' Fisher exact test 

. E ,~ "~ ~reputial Gland: Adenoma or Carcinoma ..,• 

Overall rate 7/.60(12%) 8/60 (i3%> 0/60 (0%) 8/60~/3%) 

Adjuste~f i'at.e 33.6% 34.2% 0.0% 34.2% 

Terminal rate~. , ,. 2/8 (25%) 3/14 (21%) 0/12 (0%) 3/14 (2i%) 

First incidence (days_! 381 • 520 520 

Life table test p=0.529N P=O.O04 

Logistic regression test P=0.556 P=O.O04 

Fisher exact test " P=0.500 P=O.O03 ,~. 

Prostate Gland: Adenoma 

Overall rate • 2/60 (3 %) 1/60 (2%) 3/6O (5%) 1/60 (2%) 

18:8% 7.1% Adjusted rate • . 

2/12 (17%) 1/14 (7%) Terminal rate 

712 857 (T) First incidence (days) . 

P=0.279N Life table test ' • P=0.375N Logistic regression test ~, 

P=0.309N Fisher exact test 

Skin: Keratoacanthoma 

: 4/60 (7%) , 4/60 (7%) 2/60 (3%) 4/60 (7%) Overall rate .~ 

11.0% 15.5% Adjusted rate 28.6% 15.5% 

1/14 (7%) II12 (8%) II14 (7%) Tcrmina~ rate . , 2/8 (25%) 

730 647 ': First incidence (days) 667 647 r 

• . P=0.516N P=0,326 Life table test 

P=0.616N P=0.303

P=0.641N Logistic rcgressi0fi test 

P=0.340 Fisher exact tes~ - 
~ ~~ ~ ................................. . . . . . . . . . . . . . . . . . . . . . . . . . . . ~ ~ - - ~ , ~ - ~ ~ __ ~ ...... , ...... 

Dietary II~estricfion, NT~ T~ 4~0 ~85 I ~ 

TABLE C2a 

Stafisfica~ Analysis o~ Prhnary Neopl~san s ~n Male ~ats ~n the Dietary 

Ad Libitum ]Feeding and We~ght-IV~atched Controls Protocols (continued) 

Restriction Study o~' t-~utylhy~roqu~none: 

., 

Ad LibitumFed Control 

5,000 ppm × 

Ad LibitumFed Control 

Weight-Matched 

Control 

5,0~0 ppm x 

Weight-Matched 

Control 

Skin: Squamous Cell Papilloma or Keratoacanthoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test . 

Fisher exact test 

6/60 (10%) 

45.1% 

3/8 (38%) 

667 

6/60 (10%) 

24.2% 

2/14 (14%) 

647 

P=0.423N 

P=0.566N 

P=0.619N 

2/60 (3%) 

11.0% 

1/12 (8%) 

730 

6/60 (10%) 

24.2% 

2/14 (14%) 

647 

P=0.134 

P=0.109 

P=0.136 

Skin: Trichoepithelioma or Basal Cell Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

3/60 (5%) 

.,. 

,. 

1/60 (2%) 

2.0% 

0/14 (0%) 

626 

P=0.240N 

P = 0.298N 

P =0.309N 

1/60 (2%) 1/60 (2%) 

Skin: Squamous Cell Papilloma, Keratoacanthoma, Trichoepithelioma, 

Overall rate 8/60 (13%) " 

Adjusted rate 50.3 % 

Terminal rate 3/8 (38%) 

First incidence (days) 667 

Life table test 

Logistic regression test 

Fisher exact test 

or 

7/60 (12%) 

25.7 % 

,2/14 (14%) 

626 " 

P=0.308N 

Pf0.454N 

P =0.500N 

Basal Cell Carcinoma 

3/60 (5%) 

19.0% 

2/12 (17%) 

730 

7/60 (12%) 

25.7% 

2/14 (14%) 

626 

P=0,163 

P=0.131 

P=0.!61 

Skin (Subcutaneous Tissue): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Fibroma 

~ 

: 

4/60 (7%) 

30.0 % 

1/8 (13%) 

761 

6/60 (10%) 

26.7 % 

2/14 (14%) 

642 

P=0.574 

P=0.448 

P=0.372 

7/60 (12%) 

34.3% 

3/12 (25%) 

577 

6/60 (10%.) 

26.7% 

2/14 (14%) 

642 

P=0.553N 

P=0.589N 

P=0.500N 

! 

Skin (Subcutaneous Tissue): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) . 

Life table test . • 

Logistic regression test 

Fisher exact test 

Fibrosarcoma 

1/60 (2%) 1/60 (2%) 3/60 (5 %) 

12.1% 

0/12 (0%) 

673 

1/~0 (2%) ~ 

3.6% 

0/14 (0%) 

745 

P=0.344N 

P=0.316N 

P=0.309N 

,., 

! 
186 

TABLE C2a 

Statistical Analysis of Primary Neoplasms in Male 

Ad Libitum Feeding and Weight-Matched Controls 

• , • 

Skin (Subcutaneous Tissue): Fibroma or Fibrosarcoma 

Overall rate 

Adjusted rate 

Termimd rate ' 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Testes: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test ' 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate ' • 

First incidence (days) • 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate '. 

Adjusted rate • 

Terminal rate " " " 

First.incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test ' 

Thyroid Gland (Follicular Cell): Carcinoma 

~ O~erall rate 

Adjusted :rate --

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test • 

Rats in the Dietary Restriction 

Protocols (continued) -

5,000 ppm X 

Ad Libitum- Ad LibitumFed Control Fed Control 

5/60 (8%) 7/60 (12%) 

40.0% 29~3 % 

2/8 (25%) 2/14 (14%) 

761 642 

P=0.605 

P=0.470 

P=0.381 

55/60 (92%) 59/60 (98%) 

100.0% 100.0% 

8/8 (100%) 14/14 (100%) 

489 487 

P=O.306N 

P=O.104 

P=0.103 

5/60 (8%) 4/60 (7%) 

25.2% 17.6% 

0/8 (0%) 2/14 (14%) 

730 603 

P=O.355N 

P=O.473N 

P=0.500N 

5/60 (8%) 6/60 (I0%) 

25.2% 26.5% 

0/8 (0%) 3/14 (21%) 

730 603 

P=O.575N 

P=0.536 

P=0.500 

0/60 (0%) 3/60 (5%) 

0:0% 18.8% 

0/8 (0%) 1/14 (7%) 

- 810 

P=0.237 

P=O. 179 

P=0.122 

Dietary Restriction, NTP TR 460 

Study of t-Butylhydroquinone: 

5,000 ppm × 

Weight-Matched Weight-Matched 

Control Control ~ 

9/60 (15%) 7/60 (12%) 

41.0% 29.3 % 

3/12 (25%) 2/14 (14%) 

577 642 

P=0.457N 

P=0.489N 

P=0.395N 

 ̧,, 

58/60 (97%) 59/60 (98%) 

100.0% 100.0% 

12/12 (100%) 14/14 (100%) 

577 487 

P=0.273 

P=0.258 

P=0.500 

5/60 (8%) 4/60 (7%) 

26.7% 17.6% 

2/12 (17%) 2/14 (14%) 

715 603 

P=0.495N 

P=O.564N 

P=0.500N 

6/60 00%) 6/60 00%) 

34.1% 26.5% 

3/12 (25%) 3/14 (21%) 

715 603 

P=O.605N 

P=0.535 

P=0.619N 

0/60 (0%) 3/60 (5%) 

0.0% 18.8% 

0/12 (0%) 1/14 (7%) 

- 810 

P=O.lOO 

P=0.084 . 

P=0.122 
Dietary I~es~riction, NqI~ TR 4~ ~87 

T~d~LZ C2a 

St~tLstic~ Analysis o~ ~rf~rmry Neo#~s~ns in Mane ~a~s in ~he Dietary Restriction Study o~ g-~u~ya~ydlr~one: 

Ad Libitum Feeding and Weight-Matched ControLs ~'rotocoLs (continued) ". 

Zymbal's Gland: Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Zymbal's Gland: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Mononuclear Cell Leukemia 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Malignant Mesothelioma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Benign Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Ad LibitumFFed ContrOl 

2/60 (3 %) 

9.9% 

0/8 (0%) 

500 

2/60 (3%) 

9.9% 

0/8 (0%) 

500 

39/60 (65%) 

87.2% 

4/8 (50%) 

534 

1/60 (2%) 

12.5% 

I/8 (13%) 

857 (T) 

60/60 (100%) 

100.0% 

• 8/8 (I00%) 

381 

5,000 ppm x 

Ad LibitumFed Control 

3160 (5 %) 

7.8% 

0/14 (0%) 

487 

P=0.527 

P=0.483 

P=0.500 

4/60 (7%) 

10.4% • 

0/14 (0%) 

487 

P=0.366 

P=0.321 

P=0.340 

32/60 (53%) -

75.2% 

6/14 (43%) 

423 

P=0.085N 

P =0.168N 

P=0.133N 

3/60 (5%) 

7.2% 

0/14 (0%) 

626 

P=0.366 

P=0.306 

P=0.309 

59/60 (98%) 

100.0% 

14/14 (100%) 

487 

P=0.154N 

P=0.354N 

P=0.500N 

Weight-Matched 

Contron 

1/60 (2 %) 

8.3% 

1/12 (8%) 

857 (T) 

1/60 (2%) 

8.3% 

1/12 (8%) 

857 ('IT) 

., 

45/60 (75%) 

87.9% 

7/12 (58%) 

375 

0/60 (0%) 

0.0% 

0/12 (0%) 

-

59/60 (98%) 

100,0% 

12/12 (100%) 

577 

5,~ ppm x 

Weigh~-MMtched 

Comro~ 

3/60 (5 %) 

7.8% 

0/14 (0%) 

487 

Pffi0.289 

P=0.348 

P=0.309 

4/60 (7%) [: ̧ 

10.4% 

0/14 (0%) 

487 

Pffi0.165 

P=0.213 

P=0.182 

32/60 (53%) 

75.2% 

6/14 (43%) 

423 

P=0.170N 

P =0.008N 

P=0.011N I 

3/60 (5%) 

7.2% 

0/14 (0%) 

626 

P=0.I08 

P=0.151 

-) P=0.122 

59160 (98%) 

I00.0% 

14/14 (100%) 

487 

P=0.307 

P=0.318 

P=0.752N 
188 

c 

~' ' Dietary Restriction, NTP TR 460 

TAB~I~ C2a . . ... 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of t~Butylhydroquinone: • ' • 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) ~ . 

Ad Libitum5,000 ppm X Ad Libitum- Weight-Matched 

5,000 ppm X . 

Weight-Matched 

Fed Control Fed Control Control .... Control 

• , 

All Organs: Malignant Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

51/60 (85 %) 

95.5 % 

6/8 (75 %) 

381 

• 

• 47/60 (78%) 

91.2% 

10/14 (71%) 

423 

P=0.123N 

P=0.249N 

P=0.240N 

51/60 (85%) 

93.9% 

9/12 (75%) 

375 

, . 

47/60 (78%) 

91.2% 

10/14 (71%) 

.423 • • 

P=0.507 ' 

P--0.214N 

P=0.240N .~ 

Ai! Ot~gai~: Benign 

O~/erall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

or Malignant Neoplasms 

60/60 (100%) 

100.0% 

8/8 (100%) 

381 

60/60 (100%) 

100.0% 

14/14 (100%) 

423 

P=0.180N 

6O/6O (16o%) 

100.0% 

12/12 (100%) 

375 

6O/6O (100%) 

100.0% 

14/14 (100%) 

423 " 

P=0.309 

Logistic regression test 

Fisher exact test 

f _ 

P= I:000N 

- -

P = 1.000N' ~ 

(T)Tenninal sacrifice o .... 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, liver, 

lung, pancreas, pancreatic islets, pituitary gland, preputial gland, prostate gland, testes, and thyroid gland; for other tissues, denominator is number of 

animals n¢cropsied. ~ 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjuslment for intercurrent mortality • 

Obse*ved incidence at terminal kill 

d Beneath the exposed group incidence are the P vaiheS corresponding to pairwise comparisons between the ad libitum-fed controls or weight-matched controls 

and the exposed group.' The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of:death. The 

logistic regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence 

in the exposure group is indicated by N. , • Not applicable; no neoplasms in animal group ... •

• 

f Value of statistic cannot be computed. . . 

. ~ 

: ,, ,, , 
..~ ~ 

::::-~-----:'== ~ ; :::~ . ~ , ~ . ............. , . . . . . . . . . . . ;~ .............. ~ ~:;~_ :: zz =J~ ~ =::: : ~ ................ =~ ~ 

Die~ry Restriction, N'][~ T~ 4(d) 

TABLE C2b 

Statistical. Analysis of ~?rhnary Neoplasms ~n Ma~e ~ats ~n the Dietary ]~estr~ct~on Study of t-~uty~hydroqu~none: 

~es~r~cte~ Feed Protocol " -

0 ppm 5,000 ppm 

Adrenal Medulla: Benign Pheochromocyt0ma 

Overall rate a 12/59 (20%) 12160 (20%) 

Adjusted rateb 57.5% 42.5% 

Terminal rate c 3/10 (30%) 7/22 (32%) 

First incidence. (days) 707 732 

Life table test a . P = 0.066N 

Logistic regression test d P=0.163N 

Fisher exact test d P=0.572N 

Adrenal Medulla~ Benign, Complex, or Malignant Pheochromocytoma 

Overall rate 15/59 (25 %) 14160 (23%) 

Adjusted rate 65.0 % 45.5% 

Terminal rate 4/10 (40%) 7/22 (32%) 

First incidence (days) 552 712 

Life table test P=0.037N 

Logistic regression test P=0.162N 

Fisher exact test P=0.479N 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 1/60 (2%) 3/6O (5%) 

Adjusted rate I0.0% 11.8% 

Terminal rate 1/10 (10%) 2/22 (9%) 

First incidence (days) 911 (T) 82O 

Life table test P=0.591 

Logistic regression test P=0.530 

Fisher exact test P=0.309 

Mammary Gland: Fibroadenoma 

Overall rate : .'. 4/60 (7%) 6/60 (10%) 

Adjusted rate 19.9.% 19.6% 

Terminal rate 1/10 (10%) 3/22 (14%) 

First incidence (days) 745 745 

Life table test P=0.565N 

Logistic regression test P=0.512 

Fisher exact test P=0.372 

Mammary Gland: ~'ibroadenoma or Carcinoma 

Overall rate 5/6O (8%) 7/60 (12%) 

Adjusted rate 28.8% 23.0% 

Terminal rate 2/10 (20%) 3/22 (14%) 

First incidence (days) 745 745 

Life table test P=0.463N 

Logistic regression test P=0.595 

Fisher exact .test P=0.381 

189 

[~ 

J 

i 
190 Dietary Restriction, NTP TR 460 

TABLE C2b 

Statistical .Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Restricted Feed Protocol (continued) 

•Pancreatic Islets: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Preputial Gland: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

Firstincidence (days) 

Life table test 

Logistic regression test 

Fisher exact test: 

Skin (Subcutaneous Tissue): Fibroma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): Fibroma or Fibrosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

0 ppm 5,000 ppm 

~ ~ ~ ~ "~ 

2/58 (3%) 6/60 (10%) 

8.3% 20.6% 

0/I0 (0%) 3/22 (14%) 

707 778 

~ P=0.394 

P=0.235 

P=0.147 

13/57 (23%) 16/56 (29%) 

46.6% 47.2% 

2/10 (20%) 6/20 (30 %) 

569 641 

P=0.286N 

P=0.382 

P=0.314 

2/60 (3%) 3/59 (5%) 

7.7% 8.0% 

0/I0 (0%) 0/21 (0%) 

786 731 

P=0.647N 

P=0.509 

P=0.492 

6/60 (10%) 3/60 (5%) 

33.6% 12.5% 

,1/10 (10%) 2122 (9%) 

730 866 

P=0,043N 

P=0.077N 

P~-0.245N 

' 6/60 (10%) 4/60 (7%) 

33.6% 14.0% 

, 1/10(10%) 2/22 (9%) 

730 671 

P=0.093N 

P=O. 188N 

P=0.372N 
-- 

" ~ 5~, ~ ~ 

~ d 

Dietary Rest~ct~on, ~T~ T]~ ~ 

TABLE C2b 

Statistical Ana~ys~s of ]?rhnary Heopi~sms in 1~la~e ~ts 

~estricte~ Feed ~'otoco~ (continued) 

Stomach (Forestomach): Squamous Cell Papilloma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Testes: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

in the Dietary ~est~ction 

0 ppm 

0/60 (0%) 

0.0% 

0/10 (0%) 

..~ 

59/60 (98%) 

100.0% 

10/10 (100%) 

484 

7/60 (12%) 

30.0% 

1/10 (10%) 

604 

0/60 (0%) 

0.0% 

0/10 (0%) 

7/60 (12%) 

30.0% 

1/10 (10%) 

604 

Study of t-~uty~hy~roquinone: 

$,~00 ppm 

3/60 (5%) 

9.7% 

1/22 (5%) 

743 

P=0.235 

P=0.161 

P =0; 122 

60/60 (100%) 

100.0% 

22•22 (100%) 

600 

P=0.002N 

Pffi0.600 

P=0.500 

6/59 (10%) 

17.9% 

2/22 (9%) 

730 

P = 0.191N 

Pffi0.520N 

P=0.513N 

5/59 (8%) 

12.1% 

0/22 (0%) 

641 

P=0.082 

P=0.013 

P=0.027 

11/59 (19%) 

27.8% 

2/22 (9%) 

641 

P=0.595 

P=0.158 

P=0.210 

~9~ 

i 
192 Dietary Restriction, NTP TR 460 

TABLE C2b 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Restricted Feed Protocol (continued) 

All Organs: Mononuclear Cell Leukemia 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Malignan~ Mesothelioma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Benign NeoplasmsOverallrate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact te,,;t 

All Organs: Malignant Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

0 ppm 

46/60 (77%) 

86.9% 

4/10 (40%) 

484 

3/6o (5%) 

6.0% 

O/lO (0%)- 

552 

59/60 (98 %) 

100.0% 

1o/1o (lOO%) 

484 

52/60 (87 %) 

94.0%. 

7/10 (70%) 

484 

5,000 ppm 

39/60 (65%) 

78.2% 

12/22 (55%) 

641 

P=0.002N 

P=0.234N 

P=0.114N 

4/60 (7 %) 

18.2% 

4/22 (18%) 

911 (T) 

P=0.545N 

P=0.464 

P:0.500 • 

6O16O (100%) 

100.0% 

~2/22 (100%) 

600 

P:0.002N 

P=0.6O0 

P=0.500 

49/60 (82 %) 

86.8% 

15/22 (68 %) 

600 

P=0.003N 

P=0.596N 

P=0.309N 
193 

== 

:~: ~_~.~.~~_~_._~ ......... ~ ............................................. ~ ~=~.~-~-~ ........... ~ 

Dietary Restriction,.NTP TR ~0 

TABLE C2b 

Statistical Anadys~s of Primary Neoplasms in 1V~ale Rats in the Dietary Restriction Study of t-Buty]hydroquinone: 

Restricted Feed Pr'otoeo~ (continued) 

0 ppm 5,000 ppm 

All Organs: Benign or Malignant Neoplasms 

Overall rate 59/60 (98%) 60/60 (100%) 

Adjusted rate 100.0% 100.0% 

Terminal rate 10/10 (100%) 22/22 (100%) 

First incidence (days) 484 600 

Life table test P=0.002N 

Logistic regression test P=0.600 

Fisher exact test P=0.500 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, lung, 

pancreatic islets, pituitary gland, preputial gland,, testes, and thyroid gland; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

c Observed incidence at terminal kill 

d Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the controls and that exposed group. The life table 

test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in an exposure group is indicated by 

N. 

e Not applicable; no neoplasms in animal group ' 
194 Dietary Restriction, NTP TR 460 

TABLE C3a 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of t-Butylihydroquinone: Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad Libitum. Weight-Matched 

Fed Control Control 5,000 ppm 

Disposition Summary 

Animals initially in study 70 70 70 

3-Month interim evaluation 10 10 10 

Early deaths , 

Moribund 48 41 42 

Natural deaths 4 7 4 

Survivors 

Terminal sacrifice 8 12 14 

Animals examined microscopically 70 70 70 

3-Month Interim Evaluation 

Alimentary System 

Intestine large:, colon 00) (10) (10) 

Parasite metazoan I (10%) 

Intestine large, rectum (1o) 0o) (10) 

Parasite me~azoan i (1o%) i 00%) 

Liver (1o) (1o) (10) 

Inflammation, subacute I 0o~) 2 (20%) 2 (20%) .. 

Necrosis, focal I (10%) 

Bile duct, hyperplasia 00%) 

Hcpatocyt~, vacuolization, cytoplasmic I (10%) 

Pancreas (lO) 0o) (lo) 

Atrophy 1 (I0%) 2 (20%) 

Cardiovascular System 

Heart (lO) (lO) 0o) 

Cardiomyopathy 2 (20%) 4 (4O%) 2 (20%) 

Endocrine System 

Adrenal cortex (10) 0o) (10) 

Accessory adrenal cor~qalnodule 1 (10~) 3 (30%) I (10%) 

Hyperplasia, focal l (1o%) 

Thyroid gland (10) (io) 0o) 

Ectopic thymus I (10%) i (1o%) 

Ultimobranc:hial cyst 1 (10%) 3 (30%) 

Genital System 

Prosta~ (lO) 0o) (lo) 

Inflammation, suppurative 1 (1o%) 

Number of animals examined microscopically at the site and the number of animals with lesion 

• 

. 
195 

:::=~ -~.~__~ _: .... 

. . . . . .............. ~=. =~ ::~::~ ........ :~,, , ~ ~ ................. ...... : ..... . .................. 

D~e~ry ll~estr~c!!on, N'~ T~ ~ 

T~L~ C3a 

S~nun~ry o~ the Incidence of Nenneop~as~ic Lesions ~n Mate ]Eats in the Dietary ~ees~r~ction Study 

of t-llguty~hydroquinone: Ad Libitum ~eed~ng and Weight-Matched Controls Pr'ot~co~s (continued) 

Ad Libitum- Weight-Matched 

~¢ed Control Control 5,000 ppm 

3-MOn$~ #~$¢~m ~vo~4oA~on (continued) 

Hematopoietic System 

Lymph node 

Mediastinal, hemorrhage 

Mediastinal, hyperplasia, lymphoid 

(4) 

2 (50%) 

1 (25%) 

Pancreatic, hemorrhage 2 (50%) 

Renal, hemorrhage 

Spleen 

Pigmentation, hemosiderin 

(10) 

1 (25%) 

(10) (10) 

5 (50%) 

Thymus 

Hemorrhage 

(lO) (io) 

l (lO%) 

(lO) 

l (10%) 

Respiratory System 

Lung (10) (10) (I0) 

Inflammation, subacute 4 (40%) 7 (70%) 7 (70%) 

Alveolar epithelium, hyperplasia 3 (30%) 2 (20%) 3 (30%) 

Nose (lO) (1o) (lO) 

Goblet cell, hyperplasia 7 (70%) 

Urinary System 

Kidney (1o) (I0) (I0) 

Mineralization 1 (10%) 2 (20%) 

Nephropathy 5 (50%) 8 (80%) 6 (60%) 

Systems Examined With No Lesions Observed 

General Body System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

30-Month Study 

Alimentary System 

Intestine large, colon (58) (60) (60) 

Edema 1 (2%) 

Parasite metazoan 7 (12%) 6 (10%) 2 (3%) 

Intestine large, rectum (59) (58) (59) 

Edema 2 (3%) 1 (2%) 

Hemorrhage 1 (2%) 

Parasite metazoan 5 (8%) 4 (7%) 8 (14%) 

Intestine large, cecum (60) (60) (60) 

Edema 3 (5%) 2 (3%) 3 (5%) 

Parasite metazoan 1 (2%) 2 (3%) 3 (5%) 

Intestine small, duodenum (60) (60) (60) 

Epithelium, hyperplasia 2 (3%) 4 (7%) 
196 : Dietary Restriction, NTPTR ~ 

• . 

TABLE C3a . 

Summary or the Incidence or Nonneoplastie Lesions in Male Rats in the Dietary Restriction Study 

0r t-Butylhydroquinone:, Ad Libitum Feeding and Weight-Matched Controls Protocols (conanuea) ~ ..... ., 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm • 

• ~ 

30-Month Study (continued)~ 

Alimentary System (continued) 

Intestine small, jejunum (6O) (59) (60) " 

Epithelium, hyperplasia 1 (2%) 1 (2%) 

Intestine small, ileum (60) (59) (59) 

Diverticulum I (2%) 

Liver (6o) (60) (60) 

Angiectasis 

Basophilic tbcus 

Clear cell focus 

10 (17%) 

7 (12%) 

1 (2%) 

5 (8%) 

7 (12%) 

I (2%) 

" 7 (12%). 

7 (12%) 

5 (8%) 

Congestion 

Degeneration, cystic 

Eosinophilic focus 

23 (38%) 

10 (17%) 

I (2%) 

9 (15%) 

6 (lO%) 

5 (8%) 

4 (7%) 

Eosinophilic focus, multiple ~ (2%) 

Fibrosis i (2~o) 

Hemorrhage 1 (2%) 

Hematopoietic cell proliferation 

Hepatodiaphragmatic nodule 

Mixed cell tbcus 

Necrosis, focal 

i (2~) 

7 (12%) 

2 (3%) 

8 (13%) 

4 (7%) 

, , 

6 (lO%) 

. . 

8 03%)' 

2 (3%) 

2 (3%) 

Thrombosis I (2%) 1 (2%) I (2%) 

Bile duct, cyst 

Bile duct, hyperplasia 

Centrilobular, fibrosis 

2 (3%) 

52' (87%) 47 (78%) 25 (42%) 

i (2%)- 

Centrilobular, necrosis 2 (3%) 3 (5%) 2 (3%) 

Hepatocyte, vacuolization cytoplasmic 

Kupffer cell, pigmentation 

Mesentery 

Accessory spleen 

• Fat, necrosis 

Pancreas 

Atrophy 

Acinus, cytoplasmic alteration• 

Acinus, hyperplasia, focal 

Salivary gland 

Atrophy 

Stomach, forestomach 

Edema 

6 (10%) 

11 (18%) : 

(20) 

1 (5%) 

16 (sO%) 

(60) 

15 (25%) 

3 (5%) 

2 (3%) 

(60) 

(60) 

9 (15%) 

9 05%) 

15 (25%) 

(15) 

2 (13%) 

12 (80%) 

(60) 

19 (32%) 

I (2%) 

1 (2%) 

(60) 

(2%) 

(3%) 

1 

(6O) 

2 

10 

(16) 

4 

13 

(60) 

15 

4 

2 

3 (5%). ' 

(17%) 

(25%) 

(81%) 

(25%) 

(7%) 

(3%) • 

(~o) 

(59) 

6 (1o%) " 

Erosion 2 (3%)

Hyperplasia 

Inflamm.ation, subacute 

Ulcer 

8 (13%) 

S (13%) 

(12%) 

(3%) 

(17%) 

7 

2 

I0 

1~ (20%) . 

7 ~%) 

Mucosa, hyperplasia 

Stomach, glandular 

Edema 

. Erosion 

I (2%) 

(60) 

1 (2%) 

I (2%) 

(6O) 

2 

11 

(3%) 

(1S%) 

• 

(~o) 

3 (5%) 

1 (2%) -

Mineralization 

Ulcer 

Tongue 

Epithelium, hyperplasia , 

I (2%) 

3 (5%) 

(I) , 

5 

(2) 

1 

(8%) 

(50%) .... 

".• 

(4) 

2 (50%) 

(~ I~3~, ~ " ",, ~, ~ - ~ ~.~ ,~ ~..~,,~ .~ , ~...,~ . 

,. 
!= : -~?-'~ 5~--'-:- U ~ , ~ __ _, ..... .. :,~ ....... .......................................... ..... . . . . . . . . . . 

I 

I 

r 

Dietary ll~estriction, NTP TR 460 

TABLE C3a 

Summary of the Incidence of Nonneopiastic Lesions in Male Rats in the Dietary l~estfiction Studly 

of t-~utylhydroquinone: Ad Libitum Feeding and Welght-Matched Controls ~'otocoLs (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,~ ppm 

30-Month Study (continued) 

Cardiovascular System 

Blood vessel (60) (60) (6O) 

Hypertrophy 1 (2%) 4 (7%) 

Inflammation, subacute 1 (2%) 

Heart (60) (60) (6O) 

Cardiomyopathy 40 (67%) 38 (63%) 37 (62%) 

Necrosis 1 (2%) 

Thrombosis 6 (10%) 9 (15%) 

Endocardium, hyperplasia 1 (2%) 

Schwann cell, hyperplasia 1 (2%) 

Endocrine System 

Adrenal cortex (60) (60) (60) 

Accessory adrenal cortical nodule 17 (28%) 14 (23%) 13 (22%) 

Degeneration, fatty 9 (15%) 7 (12%) 7 (12%) 

Hemorrhage 1 (2%) 3 (5%) 2 (3%) 

Hyperplasia, focal 4 (7%) I (2%) 3 (5%) 

Hypertrophy, focal 6 (10%) 5 (8%) 2 (3%) 

Necrosis 1 (2%) 1 (2%) 

Adrenal medulla (60) (60) (6O) 

Hyperplasia 26 (43%) 30 (50%) 12 (20%) 

Islets, pancreatic (60) ~ (60) (60) 

Hyperplasia 3 (5%) 3 (5%) 1 (2%) 

Parathyroid gland (55) (57) (58) 

Hyperplasia 6 (11%) 1 (2%) 9 (16%) 

Pituitary gland (60) (59) (60) 

Nuclear alteration 1 (2%) 

Pars distalis, angiectasis 4 (7%) 2 (3%) 

Pars distalis, cyst 6 (10%) 9 (15%) 7 (12%) 

Pars distalis, cyst, hemorrhagic 1 (2%) 

Pars distalis, hyperplasia, focal 10 (17%) 10 (17%) 14 (23%) 

Pars intermedia, angiectasis 1 (2%) 

Pars intermedia, cyst 

Thyroid gland 

1 (2%) 

(60) 

8 (14%) 

(60) 

2 

(60) 

(3%) 

Ultimobranchial cyst i (2%) 3 (5%) 5 (8%) 

C-cell, hyperplasia 

Follicle, cyst 

9 (15%) 

I (2%) 

6 (10%) 

1 (2%) 

7 (12%) 

General Body System 

None 

., 
198 Dietary Restriction, NTP TR 460 

TAnt~ C3a 

Sum~nary of the Incidende of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of t-Butylhydroquinone: Ad Libitutn Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

30-M'onth Study (continued) 

Genital System 

Epididymis 

Atypia cellular 

Granuloma sperm 

Preputial gland 

Cyst 

Hyperplasia 

Inflammation, chronic 

Intlanmaation, suppurative 

Prostate 

Cyst 

Fibrosis 

Inflantrnation, chronic 

Inflammation, suppurative 

Epilhelium, hyperplasia 

Testes 

Interstitial cell, hyperplasia 

Seminiferous tubule, atrophy 

Hematopoietic System 

Bone marrow 

Depletion cellular 

Hyperplasia 

Inf'dtration cellular, histiocyte 

Myelofibrosis 

Lymph ntode 

Iliac, hemorrhage 

InguiNal; hyperplasia, lymphoid 

Mediastinal, congestion 

Mediastinal, "hemorrhage 

Mediastinal, hyperplasia, lymphoid 

Mediastinal, pigmentation 

Pancreatic, hyperplasia, plasma cell 

Pancreatic, pigmentation 

Renal, ectasia 

Renal, hemorrhage ..... 

Renal, pigmentation 

Lymph node, mandibular ~.. 

Ectasia 

Hemorrhage 

Hyperplasia, lymphoid 

Pigmentation 

Lymph node, mesenterie 

Ectasia 

Hemorrhage 

Hyperplasia, lymphoid 

(6O) 

27 (45%) 

(6O) 

3 (5%) 

3 (5%) 

26 (43%) 

3 (5%) 

(60) 

1 (2%) 

3 (5%) 

5 (8%) 

36 (60%) 

11 (18%) 

(60) 

4 (7%) 

7 02%) 

(60) ,~ 

6 (10%) 

4 (7%) 

(33) 

1 (3%) 

2 (6%) 

3 (9%) 

1 (3%) 

13 (39%) 

9 (27%) 

1 (3%) ...... 

8 (24%) 

(60) 

3 (5%) 

2 (3%) 

11 (18%) 

6 (10%) 

(6O) 

7 (12%~ 

1--(2%) ............ 

3 (5%) 

(6O) 

29 (48%) 

. (6O) 

1 (2%) 

12 (20%) 

1 (2%) 

(60) 

2 (3%) 

28 (47%) 

12 (20%) 

(60) 

3 (5%) 

1 (2%). 

(60) 

I (2%) 

2 (3%) 

(33) 

3 (9%) 

2"(6%) 

14 (42%) 

4 (12%) 

......... 

5 (15%) 

(59) 

8 (14%) 

1 (2%) 

11 (19%). 

8 (14%) ' , 

(58) 

1 (2%) 

-.3 (5%) ................ 

2 (3%) 

(6O) 

25 (42%) 

1 (2%) 

(6O) 

4 (7%) 

2 (3%) 

12 (20%) 

5 (8%) 

(60) 

23 (38%) 

8 (13%) 

(6o) 

3 (5%) 

5 (8%) 

(6o) 

7 (12%) 

2 (3%) 

2 (3%),. 

(35) 

1 (3%) 

2 (6%) 

3 (9%) 

2 (6%) 

16 (46%) 

1 (3%) 

3 (9%) 

2 (6%) 

2 (6%) 

7 (20%) 

(60) 

6 (10%) 

3 (5%) 

17 (28%) 

5 (8%) 

(6o) 

10 (17%) 

2 (3%) 

6 (I0%) 
-~ 

~_- ~ ~'~'~ ~.~-- ~'7 ~...... ~------~:~ffi ~ .~ .... ..... ~=-~- . ~ .................... 

D~e~ry Restriction, NTP TS ~60 ~99 

I ~ 

i 

TAeLE C3a 

Sun~nary o~' the Incidence o~ Nonneopiasfic Les~om in MaRe Rats in the D~e~ry ~.estricfion Study 

o~ t-~uty~hydroquinone: Ad Libitum ~'eeding and Weight-IV~atched Controls ~'otoco~s (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

30-Mongh Study (continued) 

Hematopoietic System (continued) 

Spleen (60) (60) (6O) 

Angiectasis 1 (2%) 

I 

I 

Congestion 

Fibrosis 

1 (2%) 

21 (35%) 19 (32%) 13 (22%) 

Hematopoietic cell proliferation 7 (12%) 6 (10%) 8 (13%) 

i Mctaplasia, lipocyte I (2%) 

Necrosis 2 (3%) 3 (5%) • 

Pigmentation, hemosiderin 13 (22%) 11 (18%) 12 (20%) 

Lymphoid follicle, atrophy 1 (2%) 

Thymus (58) (55) (56) 

Cyst 1 (2%) 

Integumentary System 

Mammary gland (57) (58) (58) 

Dilatation 23 (40%) 27 (47%) 10 (17%) 

Galactocele 5 (9%) 2 (3%) 

. Hyperplasia 7 (12%) 12 (21%) 6 (10%) 

Skin (6O) (6O) (60) 

Cyst epithelial inclusion 2 (3%) 1 (2%) 

Hemorrhage 1 (2%) 

Hypcrkeratosis 1 (2%) 1 (2%) 

Inflammation, chronic 1 (2%) 

Epidermis, hyperplasia 1 (2%) I (2%) (2%) 

Subcutaneous tissue, inflammation 

suppurative 1 (2%) 

Subcutaneous tissue, thrombosis 1 (2%) 

Musculoskeletal System 

Bone (60) (60) (60) 

Fibrous osteodystrophy 6 (10%) I (2%) 7 (12%) . . Hyperosmsis 1 (2%) 

Femur, osteopetrosis 1 (2%) 2 (3%) 

Nervous System 

Brain (60) (60) (60) 

Angiectasis I (2%) 

Atrophy 12 (20%) 6 (10%) 3 (5%) 

Hemorrhage 2 (3%) 1 (2%) 

Hydrocephalus 4 (7%) 3 (5%) 

Necrosis I (2%) 2 (3%) 1 (2%) 

j : 

i 
200 Dietary Restriction, NTP TR 46( 

TAmE C3a 

Summm-y of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of t-Butyihydroquinone: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

30-Month Study (continued) 

Respiratory System 

Lung 

- Congestion 

Edema 

Hemorrhage 

Infdtration cellular, histiocyte 

Inflammation, subacute 

Metaplasia, osseous 

Alveolar epithelium, hyperplasia 

Nose 

Foreign body 

Inflammation, suppurative 

Goblet cell, hyperplasia 

Mucosa, hyperplasia 

Mucos~, metaplasia, squamous 

Special, Senses System 

Eye 

Atrophy 

Cataract 

• .Inflammation, chronic 

Retina, degeneration 

Urinary System 

Kidney 

Cyst 

Inflammation, suppurative 

Mineralization 

Nephropa~y 

Renal tubule, accumulation, hyaline droplet 

Renal tubule, atrophy 

Renal tubule, necrosis 

Renal tubule, pigmentation 

Transitional epithelium, hyperplasia 

Urinary bladder 

Hemorrhage . 

Inflanunation, suppurative 

• Transitional epithelium~ hyperplasia 

~_ 

~%~ 

Ad LibitumFed Control 

(60) 

2 (3%) 

1 (2%) 

3 (5%) 

19 (32%) 

1 (2%) 

4 (7%) 

(6O) 

6 (10%) 

17 (28%) 

5 (8%) 

14 (23%) 

8 03%) 

(4) 

2 (50%) 

~ (25%) 

I (25%) 

(6O) 

2 (3%) 

9 (15%) 

12 (20%) 

6O (100%) 

1 (2%) 

3 ~5%) 

18 (30%) 

13 (22%) 

(60) 

1 (2%) 

1 (2%) 

2 (3%) 

Weight-Matched 

Control . . 

(6O) 

1 (2%) 

2 (3%) 

17 (28%) 

6 (10%) " 

(60) 

5 (8%) 

10 (17%) 

4 (7%) 

8 (13%) 

4 (7%) 

(4) 

2 (50%) 

I (25%) 

2 (50%) 

(60) 

1 (2%) 

11 (18%) 

56 (93%) 

3 (5%) 

: ! (2%) 

1 (2%) 

23 (38%) 

2 (3%) 

(6O) 

1 (2%) 

1 (2%) • 

, . . 

5,000 ppm 

(6O) 

1 (2%) 

2 (3%) 

14 (23%) 

1 (2%) 

7 (12%)' 

(6O) 

2 (3%) 

11 (18%) 

13 (22%) 

10 (17%) 

7 (12%) 

(2) 

1 (50%) 

1 (50%) 

1 (50%) 

(6O) 

11 (18%) 

20 (33%) 

1 (2%) 

60 000%) 

1 (2%) 

1 (2%) 

15 (25%) 

21 (35%) 

(6O) 
~ ...... ,,----: .............. ~--.-,~__. ..... -,-- ........... -~: .......... --~=~7.-~ ~ = ~ r.,:: ::~ .... ~ ...................... ,~ ~,, 

Dietary Restriction, NTP TR 460 201 

TABLE C3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of t-Butylhydroquinone: Restricted Feed Protocol a 

0 ppm 5,000 ppm 

Disposition Summary 

Animals initially in study 70 70 

3-Month interim evaluation 10 10 

Early deaths 

Moribund 43 33 

Natural deaths 7 5 

Survivors 

Terminal sacrifice 10 22 

Animals examined microscopically 70 70 

i ~ 

3-Month Interim Evaluation 

Cardiovascular System 

Heart (10) (lO) 

Cardiomyopathy 3 (30%) 2 (20%) ! 

Endocrine System 

Adrenal cortex (10) (10) i 

Accessory adrenal cortical nodule 2 (20%) 

Pituitary gland (10) (10) 

Pars distalis, cyst I (10%) 1 (10%) 

Pars intermedia, cyst 1 (10%) !~ 

Thyroid gland (10) (10) 

Ultimobranchial cyst 2 (20%) 

Genital System 

Prostate (10) (10) 

Inflammation, suppurative 2 (20%) ~ 00%) 

! 

Hematopoietic System i 

Lymph node, mesenteric (i0) I (lO) 

Hemorrhage 1 (lO%) 

Spleen 0o) (10) 

Pigmentation, hemosiderin 3 (30%) 

Thymus (lO) (9) 

Hemorrhage 1 (10%) 

Respiratory System 

Lung (10) (10) 

Alveolar epithelium, hyperplasia 1 (10%) 

Nose (10) (lO) 

Goblet cell, hyperplasia 6 (60%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
202 Dietary Restriction, NTP TR 460 

TABLE C3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

3-Mo'nth Interim Evaluation (continued) 

Urinary System 

Kidney' 

Nephropathy 

Systems Examined With No Lesions Observed 

• Alimentary System ,, 

General Body System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

30-Month Study 

Alimentary System 

Intestine large, colon 

Parasite metazoan 

Intestine large, rectum 

Erosion 

Parasite metazoan 

Intestine large, cecum 

Edenut 

Parasite metaz0an 

Intestine small, duodenum 

Ulcer 

-Liver 

Angiectasis 

Basophilic focus 

Clear cell focus 

Cyst 

. - Degeneration, cystic 

Eosinophilic focus , 

Hemal~apoiefic cell proliferation 

Hepatodiaphragmatic nodule 

Infla~aaation, subacute 

Mixed cell focus 

Necrosis, focal 

Thrombosis 

•Bile duct, hyperplasia 

Centrilobular, necrosis 

Hepatoeyte, vacuolization cytoplasmic 

Kupffer cell, hyperplasia, 

Kupffer cell, pigmentation 

Mesentery . • • . 

Accessory spleen >. 

Fat, necrosis 

Pancreas 

Atrophy 

Acinus, cytoplasmic alteration 

of t-Butylhydroquinone: Restricted Feed Protocol (continued) 

0 ppm 

(10) 

2 (20%)' 

(60) 

7 (12%) 

(59) 

4 (7%) 

(60) 

3 (5%) 

(6(D 

3 (5%) 

(60) 

5 (8%) 

5 (8%) 

6 00%) 

6 (10%) 

I (2%) 

2 (3%) 

7 (12%) 

45 (75%~ 

2 (3%) 

l (2%) 

19 (32%) 

(8) 

2 (25%) 

4 (50%) 

(58) 

21 (36%) 

5,000 ppm 

(I0) 

1 (10%) 

(6(~ 

8 (13%) 

(C6(~ 

I (2%) 

7 (12%) 

(60) 

2 (3%) 

3 (5%) 

(60) 

(60) 

6 (10%) 

~ (2%) 

3 (5%). • 

i (2%) 

12 (20%) 

9 05%) 

5 (8%) 

i (2%) 

3 (5%) 

6 (10%) 

2 (3%) 

30 (50%) 

2 (3%) 

2 (3 %) 

14 (23%) 

(5) •

1 (2~%) 

1 (2o%) 

(~0) 

19 (32%) 

1 (2%) 
=~: ~t:~ ~--~-- ~ . ~ ~ _ . _ ~ - - - - ~ :~........ --.~:~--~. ~ ...... : ........ _+ 

[ k 

Dietm'y Restriction, NT]? T~ ~ 203 I 

T&~L~ C3b 

Su~nrnmTj o~ the l~ncfiderace o~ Nonneop~astic Lesions in Male Rats in~the Diet~ry.li~es~ric~ion S~u~Iy 

o~ t-II~uty~hydroquinone: Restricted Feedl ]Protocol (continued) 

3~-Montl~ S~u@ (continued) 

Alimentary System (continued) 

Sidivary glands 

Atrophy 

Stomach, forestomach 

Erosion 

Hyperplasia 

Ulcer 

Stomach, glandular 

Edema 

Erosion 

Mineralization 

Ulcer 

Tongue 

Epithelium, hyperplasia 

Cardiovascular System 

Blood vessel 

Hypertrophy 

Inflammation, subacu~ 

Thrombosis 

Heart 

Cardiomyopathy 

Thrombosis 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Angiectasis 

Degeneration, fatty 

Hematopoietie cell proliferation 

Hemorrhage 

Hyperplasia, diffuse 

Hyperplasia, focal 

Hypertrophy, focal 

Necrosis 

Adrenal medulla 

Hyperplasia 

Islets, pancreatic 

Hyperplasia 

Parathyroid gland 

Hyperplasia 

Pituitary gland 

Pars distalis, angieetasis 

Pars distalis, cyst 

Pars distalis, hyperplasia, focal 

Pars distalis, hypertrophy 

Pars intermedia, cyst 

Thyroid gland 

Ultimobranchial cyst 

C-cell, hyperplasia 

Follicle, cyst 

~,000 ppm 

(6O) 

1 (2%) 

(60) 

2 (3%) 

3 (5%) 

1 (2%) 

(6O) 

1 (2%) 

2 (3%) 

1 (2%) 

(2) 

t (50%) 

0 ppm 

(60) 

I 

(60) 

5 

1 

(60) 

1 

1 

1 

(1) 

(2%) 

(8%) 

(2%) 

(2%) 

(2%) 

(2%) 

( 

(60) 

1 (2%) 

1 (2%) 

1 (2%) 

(60) 

34 (57%) 

5 (8%) 

(60) 

1 (2%) 

1 (2%) 

(60) 

43 (72%) 

6 (10%) 

(60) 

13 (22%) 

I (2%) 

13 (22%) 

(2%) 

(2%) 

(3%) 

(8%) 

(59) 

18 (31%) 

(58) 

2 (3%) 

(56) 

(57) 

3 (5%) 

7 02%) 

14 (25%) 

2 (4%) 

(60) 

(8%) 

(3%) 

(60) 

7 02%) 

I (2%) 

5 (8%) 

1 (2%) 

2 (3%) 

5(8%) 

1 :(2%) 

(60) 

15 (25%) 

(6O) 

1 (2%) 

(57) 

1 (2%) 

(56) 

3 (5%~ 

7 (13%) 

7 (13%) 

1 (2%) 

4 (7%) 

(59) 

2 (3%) 

5 (8%) 

2 (3%) 
204 Dietary Restriction; NTP TR 460 

T~dn:E C3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study ' 

of t-Butylhydroquinone: Restricted Feed Protocol (continued) 

' 0 ppm 5,000 ppm 

30-Month Study (continued) 

General Body System 

None 

Genital System 

Epididymis (60) (60) 

Atypia cellular 25 (42%) 19 (32%) 

Preputial gland. (60) (59) 

Cyst 3 (5%) I (2%) 

Hyperplasia 1 (2%) 2 (3%) 

Inflammation, chronic 9 (15%) 13 (22%) 

Inflar~wnation, suppurative 1 (2%) 

Prostate (60) (60) 

Inflammation, suppurative 28 (47%) 22 (37%) 

Epithelium, hyperplasia 3 (5%) 5 (8%) 

Testes (60) (60) 

Atrophy 2 (3%) 4 (7%) 

Interstitial cell, hypgrplasia 3 (5%) 

.' 

Hematopoietic System 

Bone marrow " (60) (60) 

Hyperplasia 2 (3%) 2 (3%) 

Myelotibrosis 8 (13%) 3 (5%) 

Lymph node (36) (23) 

Iliac, hyperplasia, lymphoid .. 1 (4~) . . 

Iliac, pigmentation 2 (9%) 

Mediastinal, hemorrhage 2 (6%) 

Mediastinal, pigmentation 19 (53%) -. 11 (48%) 

Pancreatic, pigmentation 8 (22%) 4 (17%) 

Renal, hemorrhage 1 (3%) 

Renal, pigmentation 7 (19%) 3 (13%) 

Lymph node, mandibular (60) (60) 

Ectasia 2 (3%) • 2 (3%) 

Hemorrhage ! (2%) 3(5%) 

Hyperplasia, lymphoid 5 (8%) • 4 (7%) 

Hyperplasia, plasma cell 1 (2%) 2 (3%) . 

Pigmentation 13 (22 %). 14 (23%) 

Lymph node, mesenteric (60) , (60) 

Eetasia 2 (3%) 4 (7%) 

Hemorrhage 1 (2%) .. ~, 

Hyperplasia, lymphoid 1. (2%) , 2 (3~) .-

Spleen (60) (60) 

Fibrosis • . 13 (22%) ............... • " " 16 (27%)" 

Hematopoietic cell proliferation " 2 (3%) .7..,(12%)... 

Necrosis 2 (3%) 2 (3%) 

Pigmentation, hemosiderin .3. (5%) 15 (25%) 

Lymphoid foll!ele, atrophy 2 ()%) 1 (2%) / 
~ : ~-~=~¢~ , ~ , , , ~ ............ :=~o-~-~__, ..... 

Die~ry ~es~riction, ~ TR ~ 205 

TA~IL~ C3b 

S~nmry o~ the ]l~ci~ence o~ Nonneopiastic Lesioam i~ IV~aie Rats in the Die~ry ]~es~riction Study 

o~ t-]~ntyihy~roquinone: Restricte~l Fecal ]~r-otocoi (continued) 

0 ppm 5,000 ppm 

3O-Moneh S~dy (continued) 

Integumentary System 

Mammary gland 

Dilatation 

Galact0cele 

Hyperplasia 

Skin 

Cyst epilhelial inclusion 

Hyperkeratosis 

Hyperplasia 

Subcutaneous tissue, edema 

(58) 

16 (28%) 

7 (12%) 

(60) 

1 (2%) 

1 (2%) 

.. 

(58) 

14 (24%) 

1 (2%) 

9 (16%) 

(60) 

1 (2%) 

1 (2%) 

Musculoskeletal System 

Bone 

Hyperostosis 

Cranium, osteopetrosis 

Femur, osteopetrosis 

(6o) 

2 (3%) 

(60) 

1 (2%) 

1 (2%) 

2 (3%) 

Nervous System ' 

Brain 

Atrophy 

Hemorrhage 

Hydrocephalus 

Necrosis 

(60) 

3 (5%) 

I (2%) 

t (2%) 

1 (2%) 

(60) 

8 (13%) 

2 (3%) 

1 (2%) 

I 

Respiratory System 

Lung 

Congestion 

Edema 

Hemorrhage 

Inffitration cellular, histiocyte 

Inflammation, subacute 

Alveolar epithelium, hype~lasia 

Nose 

Foreign body 

Inflammation 

Inflammation, suppuratiVe 

Goblet cell, hyperp!asia 

Mucosa, hyperplasia 

Mucosa, metaplasia, squamous 

(60) 

1 (2%) 

3 (5%) 

4 (7%) 

14 (23%) 

5 (8%) 

(60) 

3 (5%) 

1 (2%) 

5 (8%) ' 

8 (13%) 

8 (13%) 

7 (12%) 

(6o) 

6 

21 

2 

8 

(60) 

2 

(10%) 

(35%) 

(3%) 

(13%) 

(3%) 

2 (3%) 

11 (18%) 

2 (3%) 

1 (2%) 

i 

I 

Special Senses System i~ 

Eye 

Cataract 

(1) 

1 (100%) 

(3) 

3'(100%) 

Hemorrhage 1 (33%) 

Retina, degeneration 1 (100%) 3 (100%) 
206 Dietary Restriction, NTP TR 460 

TABLE C3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study, 

of t-Butylhydroquinone: Restricted Feed Protocol (continued) 

• ~ . ,0 ppm 

30-Month Study (contihued) , . 

Urinary System 

Kidney (6O) 

Calculus, microscopic observation only 

Cyst 2 (3%) 

Hydronephrosis 

Infiltration cellular, iipocyte 1 (2%) 

Inflammation, suppurative 

Mineralization 4 (7%) 

Nephropathy 51 (85%) 

Renal tubule, accumulation, hyaline droplet 1 (2%) 

Renal tubule, atrophy 1 (2%) 

Renal tubule, dilatation 

Renal tubule, necrosis 3 (5%) 

Renal tubule;, pigmentation 17 (28%) 

Transitional epithelium, hyperplasia 1 (2%) 

Urinary bladder (6O) 

Transitional epithelium, hyperplasia 1 (2%) 

5,000 ppm 

(6O) 

2 (3%) 

1 (2%) 

3 (5%) 

59 (98%) 

! (2%) 

4 (7%) 

1 (2%) 

18 (30%) 

5 (8%) 

(60) 

1 (2,%) 
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I 

TABLE D~.a 

TABLE Dlb 

Summary o~ the ~nc~ence o~ ~eop~asms ~n Female ~ 

~ the D~e~y R~cfion S~e~y o~ t-~e~y~y~o~no~e: 

~d Libi~m Fee~ng and We~g~-~a~c~e~ Con~o~ ~o~o~ ................... 

Su~a~ o~ ~e ~nc~ence o~ ~eo~m~s ~n Fe~e ~a~ 

~n the Dietary ~tr~ct~on St~y o~ t-~ty~y~ro~none: 

~estr~cte~ ~ee~ ~ot~o~ ........................................... 

~0~ 

2~2 

• TABLE D2a 

T~LE D2b 

Stafisfica~ Ana~ys~ o~ ~mary ~eop~ms ~n ~e~a~e ~ 

in the Dietary ~estr~cfion Study of t-~aty~y~ro~a~none: 

Ad Lib~m ~ee~ng and We~g~t-~atc~e~ Control ~ot~o~ ................... 

Stafisfica~ Ana~ys~ of ~ary Neop~s ~a ~ema~e ~a~ 

~n the Dietary ~estr~cfion St~y o~ t-Suty~y~roqu~none: 

~estr~cte~ ~ee~ ~rotoco~ ........................................... 

2~S 

~2~ 

TABLE D3a 

TABLE D3b 

Summary o~ the ~nc~ence o~ Nonneop~fic ~o~ ~n ~e~e SaN 

~n the D~eta~ ~tr~ct~on Study o~ t-~uty~hy~roqu~none: 

Ad Libimm Feeding and We~ght-~atche~ Controls ~ot~o~ ................... 

Summary of the Incidence o~ Nonneop~c L~om ~n ~em~e ~ 

~n the D~eta~ g~tr~ct~on Study o~ t-Buty~hy~roqu~none: 

~estr~cte~ ~eed ~rot~o~ ........................................... 

~25 

23~ 
208 Dietary Restriction, NTP TR 460 

TABLE DIa 

Summary of the Incidence.of Neoplasms in Female Rats in the Dietary Restriction Study. of t-Butyihydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a . -

Ad Libitum, •.,~ .~, . . Weight-Matched 

.. ~ Fed Control Control ~ 5,000 ppm 

Disposition Summary 

Animals initially in study 70 70 70 

3-Month inteJ~n evaluation 10 10 10 

Early deaths 

Moribund 40 31 36 

Natural deaths .. 10. .7 7 

Survivors 

Terminal sacrifice I0 22 17 

Animals examined microscopically 70 70 70 

Systems Examined At 3 Months With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

Endocrine System . 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System • 

Nervous System 

Respiratory System 

Specia ! Senses System 

Urinary System 

30-Month Study . . . . . . 

Alimentary System 

Intestine large, colon 

Intestine large, cecum ~ , , 

(60) 

(60) 

(60) 

(60) 

., (59) 

(60) 

Intestine small, ileum (60) (60) (58) 

Liver (60) (60) (60) 

Fibrous histiocytoma, metastatic, skin ~ (2%) 

Hepatocellular adenoma 1 (2%) 

Hepatocellular adenoma, multiple 

Mesentery . (11 ) . (5) 

I (2%) 

(V) 

Sarcoma moraal, metastatic,.uterus 1 (9%) 

Pancreas • :.. (60) (60) (59) 

Sarcoma stromal, metastatic, Uterus I (2%) 

Salivary glands ", (60) ., (60) : ~ .. (60) 

Carcinoma I (2%) , • 

Stomach, forestomach (60) • (60) ...~ (60) 

Stomach, glandular (60) (60) (60) 

Tongue ...................................(I-) ........ (I) ..... 

Squamous cell papilloma I (I00%) 
.-~ :~: : , ~ - ~ - - , ~ - ~2 .-~ ....................... ~ .......... _ , -~- - - : = ~ - : - - ~:~::~,~.. 

Dietary Restriction, NTF TR 460 209

TABLE D~.a 

Sumrn~rry of the ~Incidence of Neoplasms inFemale Rats in the Dietary Restriction Study of t-Bdty~hydroquinone: 

Ad Libitum ]Feeding and Welght-Matched Controls Protocols (~0n~inucd) ' 

.tO-Month Study (continued) 

I Cardiovascular System [ Heart 

! Schwannoma benign 

Endocrine System 

Adrenal cortex 

Adenoma 

Adrenal medulla 

Pheochromocytoma malignant 

Pheochromocytomabenign 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Adenoma 

Carcinoma, metastatic, thyloid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars distalis, carcinoma 

Thyroid gland 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, carcinoma 

General Body System 

Peritoneum 

Genital System 

Clitoral gland 

Adenoma 

Adenoma, multiple 

Carcinoma 

Carcinoma, multiple 

Ovary 

Granulosa cell tumor benign 

Uterus 

Adcnoma 

Carcinoma 

Lciomyoma 

Polyp stromal 

Polyp stromal, multiple 

Sarcoma stromal 

Ad Libitum- Weight-Matched 

Fed ContrOl Control 

(60) (60) 

(60) (60) 

4 (7%) • 

(60) (60) 

1 (2%) 

2 (3%) 2 (3%) 

(60) (60) 

2 (3%) 1 (2%) 

(52) (55) 

2 (4%) 1 (2%) 

(60) (59) 

26 (43%) 31 (53%) 

2 (3%) 2 (3%) 

(60) (59) 

7 (12%) 6 (10%) 

1 (2%) 2 (3%) 

1 (2%) 1 (2%) 

(1) 

(58) .,. (59~ 

6 (10%) 3 (5%) .; 1 (2%) 

6 (10%) I (2%) 

I (2%) 

(6O) (60) 

1 (2%) 

(60) (60) 

., I (2%) 

6 (IO~) ~"' 9 (15%) 

I (2%) -

2 (3%) : 1 (2%) 

5,000 ppm 

• i= 

(60) 

I (2%). 

(60) 

(60) 

1 (2%) 

3,(5%) 

(59) 

2 (3%) 

(54) 

1 (2%) 

1 (2%) 

(60) 

28 (47%) 

3 (5%) 

(60) 

6 (10%) 

I (2%) 

2 (3%), 

(60) 

6 (10%) 

l (2%) 

8 (13%) 

(60) 

I (2%) 

(60) 

1 (2%)' 

1 (2%) .:. 

9 (15%) 

j 
210 Dietary Restriction~ NTP TR 460 

TAeLE Dla 

Sunnnary of the Incidence of Neoplasms in Female Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Ad L/b/turn Feeding and Weight-Matched Controls Protocols (continued) 

30-Month Study (conjugal) 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma 

Thymus 

Fibrous histiocytoma, metastatic, skin 

Integumentary System 

Mammary gla~l 

Adenoma 

Carcinoma 

Carcinoma, multiple 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Basal cell carcinoma 

Kerat0acanthoma 

Squamous cell papilloma ' 

Subcutaneous tissue, fibroma 

Subcutaneous tissue, fibrous histiocytoma 

Subcutaneous tissue, fibrosarcoma 

Subcutai~eous tissue, hemangiosarcoma 

Subcutaneous tissue, lipoma 

Subcutaneous tissue, sarcoma 

Subcutaneous tissue, schwannoma benign 

Subcutaneous tissue, schwannoma malignant 

Musculoskeletal System 

Skeletal muscle 

Fibrous histiocytoma, metastatic, skin 

Sarcoma 

Sarcoma stromal, metastatic, uterus 

Nervous System 

Brain 

Carcinoma, metastatic, mammary gland 

• Carcinoma, metastatic, pituitary gland 

Oligodendroglioma malignant 

Spinal cord 

Astrocy'toma malignant : 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

(60) (60) (60) 

(20) (14) (18) 

(59) (60) (60) 

(58) (59) (60) 

(60) (60) (60) 

I (2%) 

(56) (56) (57) 

i (2%) 

(6o) (60) (6o) 

3 (5%) 3 (5%) 2 (3%) 

8 (13%) 1 (2%) 3 (5%) 

I (2%) 

28 (47%) 21 (35%) 20 (33%) 

15 (25%) • 2 (3%) 7 (12%) 

(00) (59) (60) 

I (2%) 

I (2%) 

2 (3%) 

3 (5%) 2 (3%) 1 (2%) 

I. (2%) 

I (2%) I (2%) 

I (2%) 

I (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

(1) (1) (1) 

,,. 

I (100%) 

I (loo%) 

I (100%) 

(60) (60) (60) , 

1 (2%) 

1 (2%) 1 (2%) 

I (2%) 

(2) .. (D , (4), , 1 (25%) , 
__ __ 

~ ~ ............ : - ~ . . . . . . . . . . . - - ..... -~ --:: ...... = e~-~ __: ,7 ~-~-.!~ ......... ~ ........... . .......... : ,, ,- ~ 

Dietary I~estricti0n, NT~ TR 46{} ~'~. 

TABLE D~a 

Sunam~ry o~ the ]Incidence o~ Neoplasms in ~ema]e Rats in the Dietary ~eestr~ction Study o~ t-~uty~hydroquinone: 

Ad Libitum ~eed~ng and Weight-Matched Controls Protocols (continued) 

: Ad LibituraFed Control Weight-Matched 

Control 5,000 ppm 

30-Month Study (continued) 

RespiratorySystem 

Lung 

Alveolar/bronchiolar adenoma 

Carcinoma, metastatic, thyroid gland 

Fibrous histiocytoma, metastatic, skin 

Nose 

(60) 

2 (3%) 

(60) 

(60) 

1 (2%) 

(60) 

(60) 

1 (2%) 

1 (2%) 

(60) 

-~ 

i i~ 

Special Senses System 

None 

:i 

i =[ 

I 

Urinary System 

Kidney 

Fibrous histiocytoma, metastatic, skin 

Sarcoma stromal, metastatic, uterus 

Renal tubule, adenoma 

Urinary bladder 

Papilloma 

(60) ~ 

1 (2%) 

1 (2%) 

(59) 

(60) 

1 (2%) 

(60) 

(6o) 

(59) 

1 (2%) 

Systemic Lesions 

Multiple organs b 

Leukemia mononuclear 

Mesothelioma malignant 

(60) 

27 (45 %) 

1 (2%) 

(60) 

25 (42%) 

(60) 

27 (45%) 

Neoplasm Summary 

Total animals with primary neoplasms c 

30-Month study 

Total primary neoplasms 

30-Month study 

Total animals with benign neoplasms 

30-Month study 

Total benign neoplasms 

30-Month study 

Total animals with malignant neoplasms 

30-Month study 

Total malignant neoplasms 

30-Month study 

Total animals with metastatic neoplasms 

30-Month study 

Total metastatic neoplasms 

30-Month study 

59 

164 

52 

112 

41 

52 

2 

5 

54 

121 

45 

84 

32 

37 

2 

6 

59 

148 

50 

95 

38 

53 

4 

4 

. 

a 

b 

e 

Number of animals examined microscopically at the site and the number of animals with neoplasm 

Number of animals with any tissue examined n~icroscopically -

Primary neoplasms: all neoplasms except metastatic neoplasms 
212 Dietary Restriction, NTP TR 460 

TABLE Dlb .... 

Summary of the Incidence of Neoplasms in Female Rats in the Dietary Restriction Study of t-Butyihydroquinone: 

Restricted Feed Protocol a 

0 ppm 5,000 ppm 

Disposition SummarY 

Animals initially in study 70 70 

3-Month interim evaluation 10 10 

Early deaths 

Moribund 39 32 

Natural deaths 3 4 

Survivors 

Terminal sacrifice 18 24 

Animals examined microscopically 70 70 

Systems Examined At 3 Months With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

Endocrine System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

30-Month Study 

Alimentary ,System 

Intestine small, duodenum (59) (59) 

Intestine small, jejunum (60) (59) 

Intestine small, ileum (60) (59) 

Liver (60) (60) 

Hepatocellular adenoma 1 (2%) 

Mesentery (8) (6) 

Pancreas (60) (59)

Salivary gland,,; (60) (60) 

Schwannoma malignant 1 (2%) 

Stomach, forestomach (59) (59) 

Squamous cell papilloma 2 (3%) 

Stomach, (60) glandular (59) 

Cardiovascular System 

Heart (60) (60) 

Schwannoma 1 (2%) benign 
~ ~-~.~::~..~-~ "--y_~__..~__.~_... ............. ................ : ~ - ~ : z : :~ ~ ~ .... ~ -~: ~ ....... 

Dietary Restriction, NTP TR 4/~ 2~3 

TA~L~ Dlb 

Sumn~ary o~ the Incidence of Neoplasms in ~emage Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

II~es~ricted ]Feed Protocol (continued) 

i 30-Month Sgudy (continued) 

Endocrine System 

I Adrenal cortex 

} Adenoma 

i Adrenal medulla 

] Pheochromocytomamalignant 

[ Pheoehromocytomabenign 

~ Pheochromocytomabenign, multiple 

I Islets, pancreatic 

I Adenoma 

i Pituitary gland 

! 

•

I 

Pars distalis, adenoma 

Thyroid gland 

I C-cell, adenoma 

[ C-cell, carcinoma 

General Body System 

None 

i 

I 

Genital System 

I Clitoral gland 

Adenoma 

Carcinoma 

Ovary 

Granulosa cell tumor malignant 

Granulosa cell tumor benign 

Uterus 

Polyp stromal 

Sarcoma stromal 

Vagina 

Squamous cell papilloma 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

Thymoma benign 

Integumentary System 

Mammary gland 

Carcinoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Basal cell carcinoma 

Trichoepithelioma 

Subcutaneous tissue, fibroma 

0 ppm 

(6o) 

(57) 

3 (5%) 

1 (2%) 

(60) 

1 (2%) 

(59) 

30 (51%) 

(60) 

3 (5%) 

(59) 

3 (5%) 

2 (3%) 

(60) 

i (2%) 

2 (3%) 

(6O) 

8 (13%) 

(1) 

I OO0%) 

(60) 

(19) 

(60) 

(59) 

(60) 

(57) 

1 (2%) 

(6o) 

1 (2%) 

22 (37%) 

8 (13%) 

(6o) 

1 (2%) 

1 (2%) 

1 (2%) 

5,000 ppm 

(6o) 

1 (2%) 

(60) 

1 (2%) 

4 (7%) 

(59) 

2 (3%) 

(60) 

18 (30%) 

(59) 

6 (10%) 

1 (2%) [: 

(59) 

10 (17%) 

5 (8%) 

(60) 

1 (2%) 

(60) 

6 (10%) 

1 (2%) 

(60) 

(23) 

(59) 

(59) 

(60) 

(59) 

(59) 

16 (27%) 

1 (2%) 

(6O) 

1 (2%) 
2:14 Dietary Restriction, NTPTR 460 

TABLE Dlb 

Summary of the Incidence of. Neoplasms in Female Rats in the Dietary Restriction StUdy of t-Butylhydroquinone: 

Restricted Feed Protocol (continued) 

, ~. ,.~, 

30-Month Study (continued) 

Mnsculoskeletal System 

Skeletal muscle 

Nervous System 

Brain 

Astrocytoma malignant 

Spinal cord 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Nose 

Special Senses System 

Zymbal's gland 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs b 

Leukemia mononuclear 

Neoplasm Summary 

Total animals with primary neoplasms ¢ 

30-Month Study 

Total primary neoplasms 

30-Month Study 

Total anhnals with benign neoplasms 

30-Month Study 

Total benign neoplasms 

30-Month Study 

Total animals with malignant neoplasms 

30-Month Study 

Total malignant neoplasms 

30-Month Study • 

0 ppm 

(1) 

(6o) 

1 (2%) 

(4) 

(60) 

2 (3%) 

i (2%) 

(60) 

(1) 

1 (10o%) 

(60) 

(60) 

(60) 

37 (62%) 

59 

133 

46 

88 

40 

45 

.' 

5,000 ppm 

(6o) 

(2) 

(60) 

2 (3%) 

(60) 

(60) 

(60) 

(60) 

34 (57%) 

57 

114 

42 

71 

41 

43 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

e Primary neoplasms: all neoplasms except metastatic neoplasms 
; . ~ _ : ~ . . . . . . ! ....... ........ --'-~==---~-- ~_~ ~ --?. : ~ : 7 --~_~=~-__~1,~__ I ,; 

i 

Dietary Restriction, ~ ~ ~ 2~ i 

, 

T~ D2a 

Statistical/~aa~ys~s o~ ~Pr~ary Neoplasms in Femade RaN iin the Dietary Res~Hcfion Study o~ t-~uty~hydlroq~none: 

Ad L/b~m ]~eedl~ng and Weight-I~Iatche~ Contrro~s ~Prot~co~s 

S,~00 ppm × S,O~O ppm × 

Ad Libitum- Ad LibiL~m- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Adrenal Cortex: Adenoma 

[ Overall rate a 4/60 (7%) 0/60 (0%) 0/60 (0%) 0/60 (0%) 

i Adjusted rate b 25.9% 0.0% 

I Terminal rate c 1/10 (10%) 0/17 (0%) 

! First incidence. (days) 795 _e 

[ Life table test a P=0.019N 

I Logistic regression test d P=0.024N 

I Fisher exact test d P=0.059N 

:1i 

i Adrenal Medulla: Benign Pheochromocytoma 

i Overall rate 2/60 (3%) 3/60 (5%) 2/60 (3%) 3/60 (5%) 

i I Adjusted rate 16.4% 12.3% 7.2% 12.3% 

Terminal rate 1/10 (10%) 1/17 (6%) 1/22 (5%) 1/17 (6%) 

i 

~ 

First incidence (days) 

Life table test 

856 750 

P=0.630N 

817 750 

P=0.440 

i Logistic regression test P=0.673 P=0.480 

I Fisher exact test P=0.500 P=0.500 ~:~ 

Adrenal I~Iedulla: Benign or Malignant Pheochromocytoma 

Overall rate 2/60 (3 %) 4/60 (7%) 3160 (5%) 4/60 (7%) 

Adjusted rate 16.4% 16.9% 10.9% 16.9% 

Terminal rate 1/10 (10%) 1/17 (6%) 1/22 (5%) 1/17 (6%) 

First incidence (days) 856 750 817 750 

Life table test P=0.601 P=0.435 

Logistic regression test P=0.539 P=0.470 

Fisher exact test P=0.340 "P=0.500 

CIRoral Gland: Adenoma 

Overall rate 6/58 (10%) 7/60 (12%) 4•59 (7%) 7/60 (12%) 

Adjusted rate 38.3 % 27.0% 14.5 % 27.0% 

Terminal rate 3/10 (30%) 2/17 (12%) 2•22 (9%) 2/17 (12%) 

First incidence (days) 579 628 817 628 

Life table test P=0.375N P=O. 195 

Logistic regression test P= 0.513N P = 0.247 

Fisher exact test P=0.526 P=0.274 

Clitoral Gland: Carcinoma 

Overall rate 6/58 (10%) 8/60 (13%) 2•59 (3%) 8/60 (13%) 

Adjusted rate 43.2% 29.0% 4.0% 29.0% 

Terminal rate 4/10 (40%) 2/i7 (12%) 0•22 (0%) 2/17 (12%) 

First incidence (days) 649 750 648 750 

Life table test P=0.465N P=0.041 

Logistic regression test P=0.579N P=0.050 

Fisher exact test P=0.415 P=0.050 
216 Dietary Restriction, NTP TR 460 

TABLE D2a " 

Statistical,Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of t-Butyihydroquinone: 

Ad Eib'_.~m Feeding and Weight-Matched Controls Protocols (continued) ,. 

5,000 ppm x 5,000 ppm X 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

:.- ,. Fed Control Fed. Control Control Control 

Clitoral Gland: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Mam~nary Gland: Fibroadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Mammary Gland: Adenoma 

Overall rate 

Adjusted rate 

Tennimd rate 

First,incidence (days) .. 

Life table-test 

Logistic regression test 

Fisher exact test 

Mammary Gland: Fibroadenoma or Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test • 

Logistic regression test 

Fisher exact test 

Mammary G!and: Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test • 

Logistic regression test . 

Fisher exact test 

% 

12/58 (21%) 

74~9% 

7/10 (70%) 

579 

43•60 (72 %) 

100.0% 

. 10/10 (100%) , 

418 

3/60 (5%) 

9.9% 

0/10 (0%) 

613 

45/60 (75%) 

100.0% 

10/10 (100%) 

418 

8/60 (13%) 

29.3% 

1)10 (10%) 

540 

14/60 (23%) 

47.4% 

4/17 (24%) 

628 

P=0.248N 

P=0,402N 

P=0.451 

27/60 (45%) 

74.4%, 

9/17 (53%) 

596 

P<0.001N 

P<0,001N 

P=0.003N 

2/60 (3%) 

8.6% 

1/17 (6%) 

807 

P=0.326N 

P=0.503N 

P=0.500N 

27/60 (45%) 

74.4% 

9/17 (53%) 

596 

P<0.001N 

P<0.001N 

P<0.001N 

, . 

4/~o (7%) 

10.5% 

0/17 (0%) 

690 

P=0.070N 

P=0.177N 

P=0.181N 

6/59 (10%) 14/60(23%) 

17.9% 47,4% 

2/22 (9%) 4/17 (24%) 

648 628 

P=0.029 

P=0.040 

P=0.046 

23160 (38 %) 27160 (45%) 

62.8% 74.4% 

10/22 (45%) 9/17 (53%) 

600 -596 

P=0.157 

P=0.252 

P=0,289 

3/60 (5%) 2/60 (3%) 

8.5% 8.6% 

1/22 (5%) 1/17 (6%) 

668 807 

P=0.563N 

P=0.501N 

P=0.500N 

25/60 (42%) 27/60 (45%) 

66.6% 74,4% 

11/22 (50%) 9/17 (53%) 

600 596 

P=0.239 

P=0.389 

P=0.427 

1/~o (~%) 4/~o C7 %) 

2.1% 1o.5% 

0/22 (0%) 0/17 (0%) 

729 690 

P=0.168 

P=0.181 

P=0.182 
~ :'~---~---:~ ~-"-~'-~-----~---,_ ~ ........ ~_-7.--~ .... ...... :7::~: .... ~-~, ~--~----.~--=:~ 

Dietary ~estr~cfion, NTP Tg ~ 2~7 

TABLE D2a 

Stat~stica~ Analysis of ]?rhnary Neoplasms ~n ~ema~e Rats in the Dietary ~estr~ction Study of t-Buty~hydroqu~aoue: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

, ?. 

Ad LibitumFed Control 

5,000 ppm x 

Ad LibitumFed Control 

Weight-Matched 

Control 

5,~ ppm × 

Weight-Matched 

Control 

I 

Mammary Gland: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

10/60 (17 %) 

34.9% 

1/10 (10%) 

540 

6/60 (10%) 

18.2% 

1/17 (6%) 

690 

P=0.064N 

P=O.193N 

P=O.211N 

4/60 (7 %) 

10.4% 

1/22 (5%) 

668 

6/60 (10%) 

18.2% 

1/17 (6%) 

690 

P=0.317 

P=0.347 

P=0.372 

Mammary Gland: Fibroadenoma, Adenoma, or Carcinoma 

Overall rate 48/60 (80%) 

Adjusted rate 100.0 % 

Terminal rate 10/10 (100%) 

First incidence (days) 418 

Life table test 

Logistic regression test 

Fisher exact test 

30/60 (50%) 

76.3 % 

9/17 (53%) 

596 

P<0.0OIN 

P < 0.001N 

P < 0.001N 

26/60 (43 %) 

67.3 % 

11/22 (50%) 

600 

30/60 (50%) 

76.3 % 

9/17 (53%) 

596 

P=O.160 

P=0.264 

P=0.292 

I Pituitary Gland (Pars Distalis): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Adenoma 

26/60 (43 % ) 

80.9 % 

5/10 (50%) 

540 

28/60 (47 % ) 

69.8 % 

7/17 (41%) 

501 

P=0.103N 

P=0.481N 

P=0.427 

31/59 (53 % ) 

70.1% 

10/22 (45%) 

523 

28160 (47 %) 

69.8 % 

7/17 (41%) 

501 

P=0.537 

P=O.330N 

P=0.324N 

Pituitary Gland (Pars Distalis): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Carcinoma 

2/60 (3%) 

5.0% 

0/10 (0%) 

540 

3/60 (5%) 

10.1% 

1/17 (6%) 

666 

P=0.609 

P=0.441 

P=0.500 

2•59 (3%) 

6.5% 

1/22 (5%) 

729 

3/60 (5%) 

10.1% 

1/17 (6%) 

666 

P=0.434 

P=0.508 

P=0.508 

Pituitary Gland (Pars Distalis): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Adenoma or Carcinoma 

28160 (47%) 

81.9 % 

5/10 (50%) 

540 

31/60 (52%) 

74.0 % 

8/17 (47%) 

501 

P=0.124N 

P=0.573 

P=0.358 

33/59 (56%) 

73.1% 

11/22 (50%) 

523 

31/60 (52%) 

74.0 % 

8/17 (47%) 

501 

P=0.462 

P=O.395N 

P=0.389N 
218 Dietary Restriction, NTP TR 460 

TABLE D2a : 

Statistical Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Ad l~itum Feeding and Weight-Matched Controls Protocols (continued). 

5,000 ppm X 5,000 ppm X 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Skin (Subcutaneous Tissue): Fibroma 

Overall rate 3/60 (5%) 1/60 (2%) 

Adjusu~ ram 13.8 % 4.2 % 

Tern~tal rate 1/10 (10%) 0/17 (0%) 

First incidence (clays) 540 " 869 

Life table test P=0.200N 

Logistic regression test P=0.313N 

Fisher exact test P =0.309N 

Skin (Subcuianeous Tissue):. Fibroma, Fibrous Histiocytoma, Fibrosarcoma, or Sarcoma 

Overall rate 

Adjusted rate 

Termhtfl rate . 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid,Gland (C-cell): 

Overall~te . 

Adjusted rate 

.Tei'minal rate 

First incidence (days) ' 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): 

-O~erall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

~ Uterus: Stroma~ Polyp 

Overall rate 

Adjusted rate 

Terminal rate 

First incklence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Adenoma 

Adenoma or Carcinoma 

4/60 (7 % ) 3/60 (5 96) 

19.9% 15.4% 

1/10 (10%) 2/17 (12%) 

540 869 

P=0.279N 

P=0.410N 

P=0.500N 

7/60 (12%) 6/60 (10%) 

,64.9% 24.2% 

4/10 (40%) 3/17 (18%) 

540 633 

P=0.208N 

P=0.351N 

P=0.500N 

8/60 (13 % ) 7/60 (12 7o) 

54. 1% 27~6% 

5/10 (50%) 3/17 (18%) 

540 633 

P=0.174N 

P=0.309N 

P=0.500N 

7/60 (12%) 9/60 (15%) 

28.7% 24.8% 

1/10 (10%) 2/17 (12%) 

649 591 

P=0.554N 

P=0.388 

P=0.395 

2160 (3%) 

3/6O (5%) 

8.4% 

1/22 (5%) 

648 

6/59 (10%) 

22.5% 

3/22 (14%) 

830 

7/59 (12%) 

24.3% 

3122 (14%) 

744 

9/60 (15%) 

26.4% 

3/22 (14%) 

633 

1160 (2%) 

3/6O (5%) 

!5.4% 

2/17 (12%) 

869 

P=0.589 

P=0.654. 

P=0.660N 

6/60 (10%) 

24.2% 

3/17 (18%) 

633 

P=0.503 

P=0.592 

P=0.607N 

7/60 (12%) 

27.6% 

3/17 (18%) 

633 

P=0.498 

P=0.587 

P=0.598N 

9/60 (15%) 

24.8% 

2/17 (12%) 

591 

P=0.497 

P=0.598N 

P--0.601N 

_. 
Y~"~ Y~ :~ ;T~~ _~- -: ±: : ~ ~ ~. .............. ~ ................ ..... ~ ....... ~ - - ~ :~ ~ 

[ 

Dietary Restriction, NTP TR 460 219

TABLE D2a 

Statistical Analysis o~' Prhnary Neoplasms in Female RaN in the Dietary Restriction Study of t-~utylhydrroquinone: 

Ad Libitum Feeding and Weight-Matched Controls ]Protocols (continued) ' 

5,000 ppm × 5,000 ppm × 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Uterus: Stromal Polyp or Stromal Sarcoma 

Overall rate 9/60 (15%) 9/60 (15%) 9/60 (15%) 9/~0 (15%) 

Adjusted rate 34.9% 24.8% 26.4% 24.8% 

Terminal rate 1/10 (10%) 2/17 (12%) 3/22 (14%) 2/17 (12%) 

First incidence (days) 302 591 633 591 

Life table test P=0.332N P=0.497 

Logistic regression test P=0.536 P=0.598N 

Fisher exact test P=0.601N P=0.601N 

All Organs: Mononuclear Cell Leukemia [ 

I Overall rate 27/60 (45 %) 27/60 (45 %) 25/60 (42 %) 27160 (45 %) 

Adjusted rate 76.5% 63.2% 59,0% 63.2% 

Terminal rate 5/10 (50%) 5/17 (29%) 7•22 (32%) 5/17 (29%) 

First incidence (days) 421 501 509 501 

Life table test P=0.106N P=0.297 

Logistic regression test P=0.529N P=0.424 

Fisher exact test P=0.573N P=0.427 

All Organs: Benign Neoplasms 

Overall rate 52/60 (87 %) 50/60 (83 %) 45160 (75 %) 50160 (83 %) 

Adjusted rate 100.0% 95.8% 89.7% 95.8% 

Terminal rate 10110 (100%) 15117 (88%) 17/22 (77%) 15/17 (88%) 

First incidence (days) 418 501 523 501 

Life table test P=0.009N P=0.118 

Logistic regression test P=0.085N P=0.175 

Fisher exact test P=0.399N P=0.184 

All Organs: Malignant Neoplasms 

Overall rate 41/60 (68%) 39/60 (65%) 32/60 (53%) 39/60 (65%) 

• Adjusted rate 100.0% 82.8% 66.8% 82.8% 

Terminal rate 10/10 (100%) 10117 (59%) 8•22 (36%) 10/17 (59%) 

First incidence (days) 302 501 509 501 

Life table test P=0.024N P=0.105 

Logistic regression test P= 0.343N P-~ 0.191 

Fisher exact test P=0.423N P=0.133 
220 Dietary Restriction, NTP TR 460 

TA~I~ D2a ,-

Statistical AnalySis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) " 

5,000 ppm × 5,000 ppm × 

Ad. Libitum- Ad Libitura- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

All Organs: Benign or Malignant Neoplasms 

Overall rate 59/60 (98%) , 59/60 (98%) 54/60 (90%) 59/60 (98%) 

Adjusted rate 100.0 % 98.3 % 93.1% 98.3 % 

Terminal rate 10/10 (100%) 16/17 (94%) 18/22 (82%) 16/17 (94%) 

First incidence (days) : 302 501 509 501 

Life table test P=0.016N P=0.120 

Logistic regression test P=0.624N P=0.059 

Fisher exact test P=0.752N : P=0.057 

Number 9f neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, clitoral 

gland, pituitary gland, thyroid gland, and uterus; for other tissues, denominator is number of animals necropsied. 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence at terminal kill 

Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the ad libitum-fed controls or weight-matched controls 

and the exposed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The 

logistic regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. Forall tests~ a lower incidence 

in the exposure group i~ indicated by N. 

Not applicable; no neoplasms in animal group 
~ v .............. ___. ~-~?__~::-:~S~.~---~: .~-..:~ --~-r~ -~ ----

! 

Dietary Nestricfion, NTP TR 460 221 

i -

TAnLE D2b 

Stafistica~ Analysis of ~rimary Neoplasms in Female Rats h~ the D~e~m'y ~estr~ction Study of t-~uty~hydroqu~none: 

~estr~cted Feed Protocol ,r 

0 ppm 5,000 ppm 

i 

Adrenal Medulla: Benign Pheoehromoeytoma 

Overall rate a 

Adjusted rate b 

Terminal rate c 

First incidence (days) 

Life table test d 

Logistic regression test d 

Fisher exact test d 

4/57 (7%) 

17. 1% 

2/18 (1 ! %) 

677 

4/60 (7 %) 

12.9% 

2/24 (8%) 

499 

P=0.530N 

P=0.613N 

P=0.612N 

I 

I 

Adrenal Medulla: Benign or Malignant Pheochromocytoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

4/57 (7 %) 

17.1% 

2/18 01%) 

677 

5/60 (8%) 

15.1% 

2/24 (8%) 

499 

P=0.619 

P=0.535 

P=0.533 

Clitoral Gland: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

3/59 (5%) 

13.4% 

1/18 (6%) 

866 

10/59 (17%) 

27.5% 

3/23 (13%) 

673 

P=0.093 

P=0.044 

P=0.037 

Clitoral Gland: Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

2/59 (3%) 

6.2% 

0/18 (0%) 

788 

5/59 (8%) 

21.7% 

5•23 (22%) 

911 (T) 

P=0.310 

P=0.279 

P=0.219 

Clitoral Gland: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

5/59 (8%) 

18.8% 

1/18 (6%) 

788 

15/59 (25%) 

45.6% 

8/23 (35%) 

673 

P=0.051 

P=0.019 

P=0.013 
222 Dietary Restriction, NTP TR ~0 

TABLE D2b 

Statistical Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of t-Butyihydroquinone: 

Restricted Feed Protocol (continued) 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Mammary Gland: Fibroadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Mammary Gltand: Fibroadenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Ovary: Benign or Malignant Granulosa Cell Neoplasm 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

0 ppm 

3160 (5%) 

II.0% 

0/18 (0%) 

711 

30/60 (50%) 

86.8% 

14/18 (78%) 

688 

31/60 (52%) 

87.5% 

14/18 (78%) 

688 

3/60 (5%) 

9.7% 

0/18(0%) 

820 

30/59 (51%) 

81.1% 

-11/17 (65%) 

638 

5,000 ppm 

2/60 (3%) 

5.5% 

0/24 (0%) 

841 

P=0.402N 

P=0.490N 

P=0.500N 

17/60 (28%) 

56.1% 

12/24 (50%) 

780 

P<0.001N 

P=0.002N 

P=0.012N 

17/60 (28%) 

56.1% 

12/24 (50%) 

78O 

P<0.001N 

P < 0.001N 

P=0.0OSN 

1/60 (2%) 

4.2% 

1/24 (4%) 

911 (T) 

P=0.234N 

P=0.277N 

P=0.309N 

18160 (30%) 

50.8% 

9/24 (38%) 

649 

P=0.004N 

P=0.007N 

P=0.016N 
: - : - - ~ " - ~ " ~ 7 7 " ~ - ~ - - w - .... "-'---~---~----~ ~ " "~'~ : ~ : ' ~ - ~ " 

Die~'y Restriction, ,brT~ T~ ~6~ 

TALE D2b 

S~t~sfic~ An~ys~s o~ ]?rrhnary Iqeopimms in Female ~a~ 

]Restricted Feed~r'otocoi (continued) 

Thyroid Gland (C-cell): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Uterus: Stromal Polyp 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Uterus: Stromal Polyp or Stromal Sarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Mononuclear Cell Leukemia 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

in the Die~ry 

0 ppm 

3/60 (5%) 

11.4% 

1/18 (6%) 

677 

3/60 (5%) 

11.4% 

1/18 (6%) 

677 

8/60 (13%) 

21.2% 

1/18 (6%) 

562 

8/60 (13%) 

21.2% 

1/18 (6%) 

562 

37160 (62%) 

74.3 % 

8/18 (4.4%) 

504 

223 

~ees~riction Study o~ t-Butyihydroquinone: 

5,000 ppm 

I

i~ 

6/59 (10%) 

16.5% 

1/24 (4%) 

499 

P=0.348 

P=0.221 

P = 0.237 

7/59 (12%) 

20.1% 

2/24 (8%) 

499 

P-~0.259 

P = 0.149 

P =0.154 

6/60 (10%) 

18.5% 

2/24 (8%) 

672 

P=0.281N 

P=0.400N 

P = 0.389N 

i 

7/60 (12%) 

22.2% 

3/24 (13%) 

672 

P=0.368N 

P = 0.507N 

P =0.500N 

34160 (57%) 

64.6 % 

7/24 (29%) 

540 

P=0.178N 

P=0.552 

P=0.355N 
224 Dietary Restriction, NTP TR 460 

TABLE D2b 

Statistical Analysis of Primary Neoplasms in Female Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Restricted[ Feed Protocol (continued) : 

0 ppm 5,000 ppm ,, 

All Organs: Benign Neoplasms 

Overall rate 46/60 (77%) 42/60 (70%) 

Adjusted rate 95.6% 86.9% 

Terminal rate 16/18 (89%) 18/24 (75 %) 

First incidence (days) 562 499 

Life table test P=0.061N 

Logistic regression test P=0.189N 

Fisher exact test P=0.268N 

All Organs: Malignant Neoplasms 

Overall rate 40/60 (67 %) 41/60 (68%) 

Adjusted rate .. 76.7% 77.6% 

Terminal rate 8/18 (44%) 13/24 (54%) 

First incidence (days) 5O4 540 

Life table test P=0.293N 

Logistic regression test P=0.271 

Fisher exact test P=0.500 

All Organs: Benign or Malignant Neoplasms 

Overall rate 59/60 (98%) 57/60 (95 %) 

Adjusted ram 98.3% 96.6% 

Terminal rate 17/18 (94%) 22/24 (92%) 

First incidence (days) 504 499 

Life table test ........ P=0.098N 

Logistic regression test P=0.297N 

Fisher exact test P=0.309N 

(T)Terminal sacrifice ..- .-

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, clitoral 

gland, lung, ovary, pituitary gland, thyroid gland, and uterus; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

c Observed incidence at terminal kill 

d Beneath the exposed group incidence are the P values corresponding to pairwise compai'isons between the controls and the exposed group. The life table, 

- test regards :neoplasms in animals dying prior to terminal kill as being(directly or indirectly) the cause of death. The logistic regression testregards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in the exposed group, is indicated by 

N. 

• I 
.... ~ ~_~ S ~ . ~ ~ . ~ : ~ - ~ ..................... ~ ~q ....... ~ - - ~ . ..... . - - ............ 

Die~ry l~estricfion, N'rP TR 4~ 225 

T~d~L~ D3a 

Summary o~ the lIncidence o~ Nonneoplastic Lesions in Female ]Rats in the Dietary ]Restriction Study. 

o~ t-~utylhydroquinone: Ad Libitum Feeding and We~ght-1~tched Controls ]Protocols a : ~: 

Ad LibitumFed Control Weight-Matched 

Control 5,000 ppm 

i 

I 

I 

Disposition Summary 

Animals initially in study 

3-Month interim evaluation 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

70 

10 

40 

10 

10 

70 

70 

10 

31 

7 

22 

70 

70 

10 

36 

7 

17 

70 

3-Month ~nterim Evaluation 

Alimentary System 

Intestine large, rectum 

Parasite metazoan 

Intestine large, cecum 

Parasite metazoan 

Liver 

Angiectasis 

Hepatodiaphragmatic nodule 

(10) 

(10) 

(10) 

2 (20 %) 

(10) 

(10) 

I (10%) 

(10) 

(10) 

1 (10%) 

(10) 

(10) 

I (10%) 

1 (10%) 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Hyperplasia, focal 

Pituitary gland 

Pars distalis, cyst 

Thyroid gland 

Ectopic thymus 

Ultimobranchial cyst 

r 

(10) 

3 (30%) 

1 (10%) 

(10) 

2 (20%) 

(10) 

1 (10%) 

2 (20%) 

(10) 

I (10%) 

(I0) ,i 

(I0) 

I (10%) 

(I0) 

I (10%) 

(I0) 

. (lO) 

1 (1o%) • 

Genital System 

Clitoral gland 

Irfflammation, chronic 

Ovary 

Cyst 

Uterus 

Hydrometra 

(10) 

1 (10%) 

(10) 

(10) 

1 (10%) 

(10) 

(i0) 

(10) 

4 (40%) 

(IO) 

(lO) 

1 (lO%) 

(1o) 

2 (20%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
226 Dietary Restriction, NTP TR 460 

TABLE D3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of t-Butylhydroquinone: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

3-Month Interim Evaluation (continued) 

Hematopoietic System 

Lymph node, mandibular (10) O0) (to) 

Hemorrhage 2 (20%) 2 (20%) 

Spleen (10) (10) (to) 

Developmental malformation I (10%) 

Pigmentation, hemosiderin 5 (50%) 3. (30%) 10 (100%) 

Thymus (10) (I0) (I0) 

Hemorrhage 1 00%) 

Respiratory System 

Lung (10) (10) (10) 

Inflammation, subacute 4 (40%) 3 (30%) 2 (20%) 

Alveolar epithelium, hyperplasia 1 (10%) 1 00%) 

Nose (10) 00) 00) 

Goblet cell, hyperplasia 1 00%) 

Urinary System • 

Kidney (10) 0o) 0o) 

cyst 1 (10%) 

Mineralization 10 (100%) 10 (100%) 10 000%) 

Systems Examined With No Lesions Observed 

Cardiovascular System 

General Body System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

30-Month ;Study 

Alimentary System 

Esophagus (58) (59) (60) 

Inflammation, subacute 1 (2%) 

Intestine.large, colon (60) (60) (59) 

Edema 1 (2%) 

Parasite metazoan 6 (10%) 3 (5%) 5 (8%) 

Ulcer I (2%) 

Intestine large, rectum (59) (60) (59) 

Edema 

Parasite metazoan 

i (2%) 

2 (3%) 8 (14%) 

Intestine large, cecum 

Edema 

(60) (60) 

1 (2%) 

(60) 

2 (3%) 

Parasite metazoan 2 (3%) 1 (2%) I (2%) 
~-- ~ ~ ~ --~,~ ~,,,r,--~ ff~ ~--=~ ........ ............. ~ 7 

Dietary Restriction, ~'P TR 46~ 

TABLE D3a 

Summary o[ the ~ncidence oi~ Nonaeop~astic Lesions in Fe~nale ~ats ~ the Dietary Restriction Study 

oi~ t-Suty[hydroquinone: Ad Libitum,Feed[ng and We[ght-1~latched Controls ]Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,0~} ppm 

30-Month Smdy (continued) 

[ 

Alimentary System (continued) 

Liver (60) (60) (60) I Angiectasis I (2%) 2 (3%) I (2%) 

t

i 

Basophilic focus 

Clear cell focus 

26 (43%) 

2 (3%) 

34 (57%) 

4 (7%) 

29 (48%) 

5 (8%) 

Degeneration, cystic 1 (2%) l (2%) I (2%) 

Eosinophilic focus 10 (17%) 12 (20%) 17 (28%) 

Eosinophilic focus, multiple 1 (2%) 

Hematopoietic cell proliferation I (2%) 3 (5%) 2 (3%) 

Hepatodiaphragmatic nodule 12 (20%) 12 (20%) 18 (30%) 

Inflammation, granulomatous 12 (20%) 9 (15%) 10 (17%) 

Mixed cell focus 2 (3%) 3 (5%) 7 (12%) 

Necrosis, focal 6 (10%) 2 (3%) 3 (5%) 

Bile duct, cyst 1 (2%) 

Bile duct, hyperplasia 17 (28%) 18 (30%) 24 (40%) 

Centrilobular, atrophy I (2%) 

Centrilobular, necrosis l (2%) 1 (2%) 

Hepatocyte, vacuolization cytoplasmic 14 (23%) 7 (12%) 3 (5%) 

Kupffer cell, pigmentation 12 (20%) 9 (15%) 9 (15%) 

Mesentery (11) (5) (7) 

Accessory spleen 1 (9%) 3 (6O%) 2 (29%) 

Fat, necrosis 9 (82%) 2 (40%) 5 (71%) 

Pancreas (60) (60) (59) 

Atrophy 20 (33%) 20 (33%) 13 (22%) 

Metaplasia, hepatocyte 1 (2%) 

Acinus, cytoplasmic alteration 

Salivary glafids (6O) 

1 (2%) 

(60) 

1 

(6O) 

(2%) 

Atrophy 2 (3%) 

Cyst 1 (2%) 

Inflammation, chronic 1 (2%) 

Stomach, forestomach (6o) (60) (60) 

Edema 4 (7%) 2 (3%) 

Erosion 1 (2%) 2 (3%) 1 (2%) 

Hyperplasia 1 (2%) 

Ulcer 2 (3%) 4 (7%) 2 (3%) 

Mucosa, hyperplasia 5 (8%) 9 (15%) 5 (8%) 

Stomach, glandular (6~ (6O) (60) 

Edema 1 (2%) 1 (2%) 

Erosion 3 (5%) 1 (2%) 2 (3%) 

Ulcer 2 (3%) 4 (7%) 

Tongue 

Epithelium, hyperplasia 

~ (1) (1) 

1 (1oo%) 

Cardiovascular System 

Blood vessel (60) (60) (60) 

Inflammation, subacute 1 (2%) 

Heart (60) (60) (60) 

Cardiomyopathy 28 (47%) 35 (58%) 28 (47%) 

Thrombosis 3 (5%) 1 (2%) 

Valve, inflammation, chronic 1 (2%) , . 

~7 

227 
228 Dietary Restriction, NTP TR 460 

TABLE, D3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of t-Butylhydroquinone: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

30-Month Study (continued) 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Atrophy 

Degeneration, fatty 

Fibrosis 

• Hemorrhage 

Hyperplasia, diffuse 

Hyperplasia, focal 

Hypertrophy, focal 

Necrosis 

Adrenal medulla 

Hyperplasia 

Islets, pancreatic 

Hyperplasia 

Parathyroid gland 

Hyperplasia 

Pituitary gland 

Pars distalis, angiectasis 

Pars distalis, atypia cellular 

Pars distalis, cyst 

Pars distalis,, hyperplasia, focal 

Pars interme.dia, angiectasis 

Pars intermedia, cyst 

Thyroid gland 

Ultimobranchial cyst 

C-cell, hyperplasia 

Follicle, cyst 

Follicular cell, hyperplasia 

General Body System 

None 

Genital System 

Clitoral gland 

Cyst 

Cyst, multiple 

Fibrosis 

Hypetplasia 

Inflammation, ch~-onic 

Inflammation, suppurative 

Ovary 

Angiectasis 

Cyst , 

Uterus 

Hydrometra 

Hyperplasia, cystic 

• .. 

Ad LibitumFed Control 

(60) 

11 (18%) 

1 (2%) 

9 (15%) 

1 (2%) 

3 (5%) 

3 (5%) 

9 (15%) 

(60) 

5 (8%) 

(60) 

I (2%) 

(52) 

I (2%) 

(60) 

8 (13%) 

14 (23%) 

9 (15%) 

2 (3%) 

(60) 

2 (3%) 

8 (13%) 

1 (2%) 

(58) 

2 (3%) 

I (2%) 

3 (5%) 

2 (3%) 

2 (3%) 

(60) 

17 (28%) - 

(60) 

3 (5%) 

4 (7%) 

Weight-Matched 

Control 

(60) 

8 (13%) 

8 (13%) 

I (2%) 

6 00%) 

8 (13%) 

2 (3%) 

(60) 

7 (12%) 

(60) 

I (2%) 

(55) 

(59) 

7 (12%) 

1 (2%) 

11 (19%) 

7 (12%) 

2 (3%) 

(59) 

2 (3%) 

9 (15%) 

(2%) 

(59) 

5 (8%) 

1 (2%) 

4 (7%) 

1 (2%) 

(6O) 

1 (2%) 

9 (15%) 

(6O) 

5 (8%) 

4 (7%) 

5,000 ppm 

(60) 

9 (15%) 

12 (20%) 

1 (2%) 

1 (2%) 

2 (3%) 

7 (12%) 

(6o) 

7 (12%) 

(59) 

(54) 

1 (2%) 

(6O) 

7 (12%) 

19 (32%) 

9 (15%) 

3 (5%) 

(60) 

1 (2%) 

8 (13%) 

1 (2%) 

1 (2%) 

(60) 

5 (8%) 

4 (7%) 

2 (3%) 

1 (2%) 

(60) 

17 (28%) 

(60) 

7 (12%) 

3 (5%) 
I 

:~ ~.~-~ ~,~ ~ ~ - - ~ ........... ~ .......... C~ .... -- ....... ~ . . . . . . . = . . . . ~ *~.~::;-:~!*:'Z~ -----,=-- .:: ~?~ ~i ........... ~,+ 

Dietary Restriction, NTF TR 460 229

TA~L~ D3a 

Surnrnary of the Incidence of Nonneopl~st[c Lesions in Female Rats in the Dietary R~triction Study 

of t-~utylhydroquinone: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

30-Month Study (continued) 

Hematopoietic System 

Bone marrow 

Depletion cellular 

Fibrosis 

Hyperplasia 

Myelofibrosis 

Stromal cell, hyperplasia 

Lymph node 

lliac, hemorrhage 

Iliac, hyperplasia, lymphoid 

Mediastinal, ectasia 

Mediastinal, hemorrhage 

Mediastinal, hyperplasia, lymphoid 

Mediastinal, pigmentation 

Pancreatic, pigmentation 

Renal, hemorrhage 

Renal, pigmentation 

Lymph node, mandibular 

Eetasia 

Hemorrhage 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Pigmentation 

Lymph node, mesenteric 

Ectasia 

Hemorrhage 

Hyperplasia, lymphoid 

Pigmentation 

Spleen 

Fibrosis 

Hematopoietic cell proliferation 

Hemorrhage 

Hyperplasia, lymphoid 

Metaplasia, lipocyte 

Necrosis 

Pigmentation, hemosiderin 

Lymphoid follicle, atrophy 

Red pulp, atrophy 

(60) 

1 (2%) 

1 (2%) 

2 (3%) 

(20) 

1 (5%) 

3 (15%) 

1 (5 %) 

14 (70%) 

1 (5%) 

1 (5%) 

(59) 

2 (3%) 

3 (5%) 

13 (22%) 

1 (2%) 

20 (34%) 

(58) 

2 (3%) 

2 (3 %) 

(60) 

3 (5%) 

15 (25 %) 

24 (40%) 

1 (2%) 

1 (2%) 

(60) 

I (2%) 

1 (2%) 

' I (2%) 

(14) 

I (7%) 

I (7%) 

(7%) 

(21%) 

(14%) 

3 (21%) 

(6~ 

2 (3%) 

3 (5%) 

12 (20%) 

27 (45%) 

(59) 

4 (7%) 

4 (7%) 

1 (2%) 

(60) 

4 (7%) 

12 (20%) 

28 (47%) 

1 (2%) 

(60) 

I (2%) 

3 (5%) 

(18) 

6 (33%) 

4 (22%) 

2 (11%) 

7 (39%) 

(60) 

1 (2%) 

7 (12%) 

1 (2%) 

25 (42%) 

(60) 

2 (3%) 

7 (12%) 

4 (7%) 

1 (2%) 

(60) 

3 (5%) 

11 (18%) 

i (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

41 (68%) 

1 (2%) 

! 

hategumentary System. 

Mammary gland (60) (60) (60) 

Dilatation 37 (62%) 33 (55%) 34 (57%) 

Galactocele 5 (8%) 1 (2%) 5 (8%) 

Hypcrplasia 12 (20%) 12 (20%) 15 (25%) 

Inflammation, suppurative 1 (2%) 

Skin (60) (59) (60) 

Cyst epithelial inclusion 1 (2%) 1 (2%) 

Hyperkcratosis 1 (2%) 1 (2%) 

Inflammation, chronic I (2%) 2 (3%) 

Ulcer 2 (3%) 2 (3%) 

Epidermis, hyperplasia 1 (2%) 2 (3%) 2 (3%) 

I 
230 Dietary Restriction, NTP TR 460 

'I'Ama~ D3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of t-Butyihydroquinone: AdlLibitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

30-Month Study (continued) 

Muscaloskeletal System 

Bone (6O) (6o) (6o) 

Fibrous osteody~trophy I (2%) 

Hy~.'rostosis I (2%) 

Cranium, osteopetrosis 8 (13%) 17 (28%) 9 (15%) 

Femur, osteopetrosis 9 (15%) 17 (28%) 5 (8%) 

Nervous System 

Brain (6o) (6o) (6o) 

Atrophy 19 (32%) 22 (37%) 22 (37%) 

Gliosis 1 (2%) 

Hemorrhage 1 (2%) 

Hydrocephalus 4 (7%) 4 (7%) 8 (13%) 

Necrosis 1 (2%) 

Respiratory System 

Lung (60) (60) (60) 

Congestion 

Edema 

1 (2%) 

2 (3%) 

Hemorrhage 

Infiltration cellular, histiocytc 

2 

28 

(3%) 

(47%) 

I (2%) 

33 (55%) 

2 (3%) 

23 (38%) 

Inflarnmation, suhaeute 3 (5%) 1 (2%) 

Thrombosis 1 (2%) 

Alveolar epithelium, hyperplasia 

Nose 

4 

(60) 

(7%) 6 (10%) 

(60) 

4 (7%) 

(60) 

Foreign body 

Inflammation, suppurative 

Goblet cell. hyperplasia 

Mucos, a, hyperplasia 

3 

5 

10 

5 

(5%) 

(8%) 

(17%) 

(8%) 

1 (2%) 

3 (5%) 

8 (13%) 

1 (2%) 

3 .(5%) 

6 (10%) 

1 (2%) 

Mucosa, metaplasia, squamous 3 (5%) 2 (3%) 

Special Senses System 

Eye 

Cataract 

(4) 

3 (75%) 

(1) 

1 (1oo%) 

Hemorrhage 

Retina, degeneration 

1 (25%) 

4 (1oo%) ~ 000%) 
'-~-"~---'~'~-~-__,~! .... .......... ~,.............................. ~ - ,~ .._ :- = .: : .......... 

Dietmry Restrictions NTP TR 460 231 } 

{ 

T~ D3a 

Summary of the lInddence of Nonneop~sfic Lesions in Female ]Rats in the Dietary gestr~ct~on Study 

of t-~uty~hydroquinone: Ad Libitum Feedingand Weight-lVi[atched Controls ]Pr-otoco~s (continued) 

Ad LibitumFed Control 

Weight-Matched 

Control 5,000 ppm 

{ 

: { 

{ 

{I 

: { 

i [ 

;[ 

[ 

: { :! 

i 

30-Month S~dy (continued) 

Urinary System 

Kidney 

Cyst 

Hydronephrosis 

Inflammation, chronic 

Inflammation, suppurative 

Mineralization 

Nephropathy 

Renal tubule, atrophy 

Renal tubule,cytoplasmic alteration 

Renal tubnie, dilatation 

Renal tubule,necrosis 

Renal tubule, pigmentation 

Transitional epithelium, hyperplasia 

Urina~ bladder 

Transitional epithelium, hyperplasia 

(6O) 

1 (2%) 

2 (3%) 

57 (95%) 

37 (62%) 

1 (2%) 

5 (8%) 

2 (3%) 

15 (25%) 

2 (3%) 

(59) 

(6O) 

1 (2%) 

I (2%) 

I (2%) 

52 (87%) 

38 (63%) 

1 (2%) 

3 (5%) 

I (2%) 

9 (15%) 

4 (7%) 

(60) 

{ (2%) 

(6O) 

2 (3%) 

5 (8%) 

48 (8O%) 

39 (65%) 

5 (8%) 

2 (3%) 

I (2%) 

2 (3%) 

16 (2'7%) 

3 (5%) 

(59) 

2 (3%) 

[ 

~+ [ 

Y 
232 Dietary Restriction, NTP TR 460 

TABLE D3b 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in ,the Dietary Restriction Study 

of t-Butyihydoquinone: Restricted Feed Protocol a 

0 ppm 5,000 ppm 

Disposition Summary 

Anim~ds initially in study 70 70 

3-Month interim evaluation I0 10 

Early deaths . 

Moribund 39 32 

Natural deaths 3 4 

Survivors 

Terminal sacrifice 18 24 

Animals examined microscopically 70 70 

3~Month Interim Evaluation 

Alimentary System 

Intestine large, colon (10) (lO) 

Parasite metazoan 1 00%) 

Intestine large, rectum (lO) (10) 

Parasite metazoan 1 (lO%) 

Liver (10) (lO) 

Hepatodiaphragmatic nodule 2 (20%). 

Cardiovascular System 

Heart (10) (~o) 

Cardiomyopathy '\ 1 (10%) 

Endocrine System 

Adrenal cortex (10) (10) 

Accessory adrenal cortical nodule 1 (10%) 

Thyroid gland (10) (io) 

Ectopic thymus i (lO%) 

Ulfimobranchial cyst i (10%) 

Genital System 

Clitoral gland (10) (lO) 

Cyst 1 (lo%) 

Ovary (10) (lO) 

Cyst 2 (20%) 

Uterus (10) (10) .. .

Hydrometra 1 (10%) 

Hematopoietic System 

Spleen (lO) (10) 

Pigmentation, hemosiderin 8 (80%) 

Number of animals examined microscopically at the site and the number of animals with lesion 
~;7-" : -_~,~.:~-:~_~-~ " ~ - - ~ ~ _ . ~ _ _ : ~: ~ . ~__ .......... ~ ..................... . .......... 

Dietary ~estriction, NTP T~ 460 233 

TA~.~ D3b 

S~'yj o~ the 1Incidence o[ Nonneop~stlc Lesions in Female Rats in the Dietary ~estrictiora Study 

o~ t-~uty~ydoquinone: Restricted Feed ]?rotocol (continued) 

0 ppm 5,000 ppm 

3-Month ~nger~m Evaluation (continued) i Respiratory System 

Nose (10) (lO) 

Goblet cell, hyperplasia 4 (40%) ? 

7 

Urinary System I 

Kidney 

Mineralization 

Systems Examined With No Lesions Observed 

General Body System I ~ntegumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

30-Month Study 

Alimentary System 

Esophagus 

Epithelium, hype~lasia 

Intestine large, colon 

Parasite metazoan 

Intestine large, rectum 

Parasite metazoan 

Intestine large, cecum 

Edema 

Parasite metazoan 

Intestine small, duodenum 

Erosion 

Epithelium, hyperplasia 

Liver 

Angiectasis 

Basophilic focus 

Clear cell focus 

Cyst 

Eosinophilic focus 

Hematopoietic cell proliferation 

Hepatodiaphragmatic nodule 

Inflammation, granulomatous 

Inflammation, subacute 

Mixed cell focus 

Necrosis, focal 

Bile duct, hyperplasia 

Centrilobular, necrosis 

Hepatocyte, vacuolization cytoplasmic 

Kupffer cell, pigmentation 

Mesentery 

Accessory spleen 

Fat, necrosis 

(10) 

10 (100%) 

(59) 

(60) 

6 (10%) i 

(60) 

6 (10%) 

(59) 

1 (2%) 

(59) 

(60) 

5 (8%) 

23 (38%) 

2 (3%) 

1 (2%) i 

13 (22%) 

3 (5%) 

3 (5%) 

1 (2%) 

2 (3%) 

24 (40%) 

1 (2%) 

9 (15%) 

19 (32%) 

(6) 

5 (83%) 

(58) 

1 

(59) 

4 

(60) 

5 

(60) 

1 

1 

(59) 

1 

1 

(60) 

26 

2 

7 

2 

2 

7 

1 

8 

7 

23 

7 

14 

(8) 

2 

5 

(10) 

10 (100%) 

(2%) 

(7%) 

(8%) 

(2%) 

(2%) 

(2%) 

(2%) 

(43%) 

(3%) 

(12%) 

(3%) 

(3 %) 

(12%) 

(2%) 

(13%) 

02%) 

(38%) 

(12%) 

(23%) 

(25%) 

(63%) 

i 
234 Dietary Restriction, NTP TR 460 

'I'A~.LE D3b 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the Dietary Restriction Study 

of t-Butylhydoquinone: Restricted Feed Protocol (continued) 

" 0 ppm 5,000 ppm 

30-Month Study (continued) 

Alimentary System (continued) 

Pancreas (60) (59) 

Atrophy 15 (25%) 11 (19%) 

Acinus, cytoplasmic alteration 1 (2%) 1 (2%) 

Stomach, forestomach (59) (59) 

Edema 1 (2%) 1 (2%) 

Erosib~ 1 (2%) 2 (3%) 

Ulcer 4 (7%) 

Epithelium, hyperplasia 5 (8%) 5 (8%) 

. Stomach, glandular (59) (60) 

Edema 1 (2%) 

Erosion 1 (2%) 2 (3%) 

Mineralization 1 (2%) 

Ulcer' 2 (3%) 

Tongue (3) 

. Epithelium, hyperplasia ' 1 (33%) 

Cardiovascular System 

Heart (60) (60) 

.Cardiomyopathy 29 (48%) 30. (50%) 

Mineralization I (2%) 

Thrombosis 2 (3%) 2 (3%) 

Endocardium, hyperplasia 1 (2%) 

Schwann cell, hyperplasia 1 (2%) 

Endocrine System 

Adrenal cortex •

Accessory adrenal cortical nodule 

Atrophy 

Degeneration, fatty 

Hemorrhage 

Hyperplasia, diffuse 

Hyperplasia, focal 

Hypertrophy, focal 

Necrosis 

Adrenal medulla 

Hype~lasia 

Islets, pm~creatic 

Hyperplasia 

Parathyrnid gland 

Hyperplasia 

Pituitary gland .: ,. 

Pars-distalis, angiectasis 

Pars distalis, cyst 

Pars distalis, hyperplasia, focal 

Pars intermedia, angiectasis 

Pars interrnedia, cyst 

(60) 

9 (15%) 

II (18%) 

I (2%) 

I (2%) • 

7 (12%) 

5 (8%) 

I (2%) 

(57) 

5 (9%) 

(60) 

(55) 

2 (4%) 

(59) 

2 (3%) 

9 (15%) 

i1 (19%) 

2 (3%) 

4 (7%) 

(6~ 

I0 

,2 

I0 

(6O) 

15 

(59) 

I 

(57) 

(6o) 

10 

11 

13 

4 

6 

(17%) 

(3%) 

(17%) 

(2%) 

(13%) 

(25%) 

(2%) 

(17%i 

(18%) 

(22%) 

(7%) 

(lO%) 
~ : = ~ : ~ = 7 = - - - ~ ~ ~ . ~ ~_ ~ .... ::= = ............ : .... ~ ~ : ~-~-~ . . . . . . . . . . . . . 

Die~-y Res*riction, ~TP TR ,~0 

T~L~ D3b 

S~n~ o~ ~he Ilncidence o~ Nonneo#mtic Lesions in Fem~fle Rats in ~he ~ie~y Res~cfio~ SOurly 

ot t-ll~u~y~ydoqulnone: Res~tric~ted Feed Pro~tocol (continued) I 

i 

0 ppm 5,~ ppm 

30-Montl~ Study (continued) 

[ Endocrine System (continued) 

I Thyroid gland (60) (59) 

Ultimobranchial cyst 2 (3%) 3 (5%) 

C-cell, hyperplasia 4 (7%) 3 (5%) 

I Follicle, cyst 1 (2%) I (2%) I 

i 

I 

I General Body System 

Ii None 

1 

~1 

Genital System 

Clitoral gland (59) (59) 

Cyst 5 (8%) 5 (8%) 

i 

Hyperplasia 

Inflammation, chronic 

5 (8%) 

3 (5%) 

6 (10%) 

2 (3%) 

Inflammation, suppurative 2 (3%) 

Ovary (60) (60) 

Angiectasis 2 (3%) 

Cyst 6 (10%) 12 (20%) 

Uterus (60) (60) 

Hydrometra 4 (7%) 3 (5%) 

Hyperplasia, cystic 1 (2%) 

Hematopoietic System 

Bone marrow (60) (60) 

Hyperplasia 1 (2%) 

Infiltration cellular, histiocyte 

Inflammation, granulornatous 

Myelofibrosis 

Stromal cell, hyperplasia 

Lymph node 

Iliac, ectasia 

Iliac, hemorrhage 

Iliac, pigmentation 

Mediastinal, hemorrhage 

Mediastinal, hyperplasia, lymphoid 

Mediastinal, pigmentation 

Pancreatic, ectasia 

I (2%) 

I (2%) 

(19) 

I (5%) 

8 (42%) 

I (5%) 

I (2%) 

I (2%) 

5 (8%) 

I (2%) 

(23) 

I (4%) 

I (4%) 

3 (13%) 

I (4%) 

11 (48%) 

Pancreatic, pigmentation 

Renal, pigmentation 

Lymph node, mandibular 

Ectasia 

Hemorrhage 

2 (II%) 

6 (32%) 

(60) 

3 (5%) 

5 (8%) 

4 

5 

(59) 

2 

5 

(17%) 

(22%) 

(3%) 

(8%) 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Pigmentation 

9 (15%) 

1 (2%) 

22 (37%) 

3 

1 

22 

(5%) 

(2%) 

(37%) 
236 Dietary Restriction, NTP TR 460 

TABLE D3b ' ,... ~

Summary of the Incidence of Nonneoplastic Lesions in Female Ratsin the Dietary Restriction Study 

of t-Butylhydoquinone: Restricted Feed Protocol (continued) " ' ' 

30-Month Study (continued) 

Hematopoietic System (continued) 

Lymph node• mesenteric 

Ectasia 

Hemorrhage. 

Hyperplasia lymphoid • :Y. • 

Plgmemanon . 

Spleen 

Fibrosis 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

Necrosis 

Pigmentation, hemosiderin 

Lymphoid follicle, atrophy 

Red pulp, atrophy 

Integumentary System 

Mammary gland 

Dilatation 

Galactocele 

Hyperplasia 

Skin 

Inflammation, chronic 

Ulcer 

Epidermis, hype~lasia 

Subcumneous tissue, edema 

Musculoskeletal System 

Bone 

Fibrous osteodystrophy 

Cranium, osteopetrosis 

Femur, osteopetrosis 

. . 

Nervous System 

Brain 

Atrophy 

Gliosis 

Hemorrhage 

Hydrocephalus 

Respiratory System 

Lung 

Edema 

Fibrosis 

Foreign body 

Hemorrhage 

Infiltration cellular, histiocyte 

Inflammation, subacute 

Mineralization 

Alveolar epithelium, hyperplasia 

. • 

0 ppm 

(59) 

1 (2%) 

2 (3%) 

1 (2%) 

(60) 

4 (7%) 

9 (15%) 

1 (2%) 

30 (50%) 

(6o) 

.51 (85%) 

1 (2%) 

3 (5%) 

(60) 

1 -(2%) 

1 (2%) 

1 (2%) 

( 

(6~ 

2 (3%) 

3 (5%) 

3 (5%) 

(60) 

17 (28%) 

1 (2%) 

1 (2%) 

(60) 

1 (2%) 

1 (2%) 

2 (3%) 

33 (55%) 

1 (2%) 

I (2%) 

3 (5%) 

5,000 ppm 

: 

(59) 

2 (3%) 

-6 (10%) 

I (2%) 

(6O) 

2 (3%) 

12 (20%) 

I (2%) ~ 

1 (2%) 

30 (50%) 

2 (3%) 

I (2%) 

(59) 

.34 (58%) 

4 (7%) 

2 (3%) 

(60) 

2 (3%) 

2 (3%) 

3 (5%) 

3 (5%) 

(60) 

2 (3%) 

7 02%) 

(60) 

15 (25%) 

I (2%) 

3 (59~) 

(60) 

3 (5%) 

I (2%) 

3 (5%) 

30 (50%) 

4 (7%) 
I 

= ~-~,~ ..... ~-~___~. ~ - ~ _. . ~ ~ .~ ......... =~:- --~._.o_ -~..~- ......... ~_=-~...~ 

, Dietary Restriction, NTP TR 460 

TABLE ~3b 

S~m~ry o~ the ]Incidence of Nonnee~pl~s~ic Lesions in Ferule ~s 

o~ t-~ntyihydoqu~none: Restricted Feed Protocol (continued) 

0 ppm 

30-Month Study (continued) 

~espiratory System (continued) 

Nose (60) 

Foreign body 1 (2%) 

Inflammation, suppurative 3 (5%) I Goblet cell, hyperplasia 7 (12%) 

Mucosa, hyperplasia 4 (7%) 

Mucosa, metaplasia, squamous 1 (2%) 

Special Senses System l

i Ear (2) . 

Inflammation, chronic 1 (50%) 

Eye 

Cataract 

Retina, degeneration 

Urinary System 

Kidney (60) 

Glomerulosclerosis 1 (2%) 

Hydronephrosis 

Inflammation, chronic 

Inflammation, suppurative 

Mineralization 59 (98%) 

Nephropathy 38 (63%) 

Renal tubule, atrophy 1 (2%) Renal tubule, cytoplasmic alteration 5 (8%) Renal mbnie, dilatation 2 (3%) 

Renal tubule, necrosis 1 (2%) 

Renal tubule, pigmentation 17 (28%) 

Transitional epithelium, hyperplasia 5 (8%) 

Urinary bladder (6O) 

Transitional epithelium, hyperplasia 1 .(2%) 

237 

in ~he Die~ry ~es~ricfion Stndy 

5,~00 ppm 

(60) 

1 (2%) 

2 (3%) 

9 (15%) 

3 (5%) 

1 (2%) 

(1) 

1 (100%) 

1 (100%) 

(6o) 

2 (3%) 

2 (3%) 

1 (2%) 

56 (93%) 

41 (68%) 

14 (23%) 

1 (2%) 

7 (12%) " 

1:~ (28%) 

5 (8%) 

(6O) 
238 

Dietary Restriction, NTP TR 460 
239 

~ - 4"~--- ~ " _ .... ~ .... ~ .......... ~ ~ w ~ 

AI~P~NI2,I~X X 

5~IA.IXY ©I~ ]LI~5]I©N5 I~N 1ViI.AIL]E ~AT5 

~N TI~--I~...~ ~I~XT.AXY IB~STX~ICT]ION 5TUDY 

@IF 5AIL~ICYILAZ©S~IL~FAI~XY~]I~I[N]E 

J TABLE E~a Suanm~ry of ~e ]~cl~e~ce of Neoplasms i~ Male ~ts 

un the D~e~ary ~r~c~o~ S~e~y o~ Saticy~zosu~p~e: 

~d Lib~m Fee~g ~ We~gh~-~che~ Con~ro~ ~o~o~ ..... -.............. 2~ 

TABLE ~Lb S~ o~ ~e ~c~e~ce o~ ~eop~m~s ~n ~a~e ~a~ 

nn the D~e~ry ~tr~ct~on Study of Sa]~cy~osu]fap~ne: 

2-Year and 30-Month Rmtr~cte~ ~eed ~otoco~ ............................ 2~5 

TABLE ~2a Stat~ca~ Ana~ys~ o~ Primly Neoplmms in MaZe RaN 

~n the Dietary ~tr~ct~on Study of Sa~cy~azosu~apyr~ne: 

Ad L&~m Feeding and Weight-Watched Controls ~ot~o~ ................... 250 

TABLE ~2b Stat~st~ca~ Ana~ys~ o~ Primary Heoplmms ~n ~ale ~ 

I 

~n the D~eta~ ~tr~ct~on Study of S~cy~osu~apyr~d~ne: 

2-fear and 30-Month ~r~cte~ ~ee~ ~ot~o~ ............................ 25~ 

TABLE ~3~ Summary of the ~nc~ence o~ Honneop~mt~c L~om in ~e ~ 

~n the ~eta~ ~tr~ct~on Study o~ Sa~cy~azosu~fapyr~d[ne: 

Ad L&i~m ~ee~ng and Weight-Hatched Control ~ot~o~ ................... 2~3 

TABLE ~3b S~ma~ o~ the ~nc[dence of Honneop~mt~c L~om ~n ~e ~a~ 

~ the D~eta~ ~tr~ct~on Study of Sa~cy~azosu~apyr[d~ne: 

2-Year and 30-Month ~r~cte~ ~ee~ ~ot~o~ ............................ 271 
240 Dietary Restriction, NTP TR 460 

TABLE Ela • 

Summa~, of the Incidence Of Neoplasms in Male Rats in the ~Dietary Restriction study of Salicylazosnlfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad LibitutnFed Control 

Weight-Matched 

Control ~ 337.5 mg/kg 

Disposition Summary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Accidental deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

• 

70 b 

i 0 

1 

13 

1 

35 

60 

10 

2 

16 

1 

31 

60 

10 

2 

15 

10 

23 

Animals examined microscopically 60 

15-Month Interim Evaluation 

Alimentary System 

Intestine large, colon 

Carcinoma 

(10) 

1 (10%) 

(lO) (~o) 

Endocrine System 

Islets, pancreatic 

Adenoma 

Pituitary gland 

Pars distalis, adenoma 

(10) 

(10) 

1 (10%) 

(10) 

1 (10%) 

(lO) 

4 (40%) 

0o) 

0o) 

Genital System 

Testes • 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

(10) 

6 (60%) 

3 (30%) 

(lO) 

3 (30%) 

5 (50%) 

(lO) 

9 (90%) 

~ 00%) 

Respiratory System 

Lung 

Alveolar/bronchiolar carcinoma 

(10) 

1 (10%) 

0o) (lO) 

' "' C 

Systems Examined With No Neoplasms Observed 

Cardiovascular System 

General Body System .... 

Hematopoietic System 

Integumentary System 

Musculoskeletal System • 

Nervous System ., 

Special Senses System ..... 

Urinary System 
~: ~ ~ "~ '~--~'~ ~. _~ ~ , ~ ~ ~ ............ ,, ~ ..... ,r ~ ~: :: _ . ~ ~ ~ :~ 5~" ~r~q, ~ ...... 

Dietary Restriction, NTP TR 460 

T~d~LE E~a i 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of Sag~cy~azosulfapyrid~ne: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

~ 7 i 

Fed Control Control 337.5 mg/kg 

i 

I 

I 

2-Year Study 

Alimentary System 

Intestine large, colon 

Adenoma 

Intestine large, rectum 

Polyp 

Intestine large, cecum 

Sarcoma 

Intestine small, jejunum 

Intestine small, ileum 

Leiomyosarcoma 

Liver 

Fibrous histiocytoma, metastatic, skin 

Hepatocellular adenoma 

Leiomyosarcoma, metastatic, stomach, 

forestomach 

Osteosarcoma, metastatic, bone 

Sarcoma, metastatic, intestine large, cecum 

Mesentery 

Fibrous histiocytoma 

Leiomyosarcoma, metastatic, stomach, 

forestomach 

Sarcoma, metastatic, intestine large, cecum 

Pancreas 

Fibrous histiocytoma 

Leiomyosarcoma, metastatic, stomach, 

. forestomach 

Mixed tumor benign 

Sarcoma, metastatic, intestine large, cecum 

Acinar cell, adenoma 

Acinar cell, adenoma, multiple 

Acinar cell, carcinoma, multiple 

Salivary glands 

Adenoma 

Schwannoma malignant 

Stomach, forestomach 

Leiomyosarcoma 

Squamous cell carcinoma 

Squamous cell papilloma 

Tongue 

Squamous cell carcinoma 

(49) 

(50) 

(50) 

(50) 

(49) 

1 (2%) 

(50) 

2 (4%) 

1 (2%) 

(17) 

1 (6%) 

1 (6%) 

(50) 

1 (2%) 

i (2%) 

9 (18%) 

3 (6%) 

1 (2%) 

(50) 

(50) 

i (2%) 

1 (2%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

(50) 

(50) 

2 (4%) 

I (2%) 

(16) 

1 (6%) 

(50) 

I (2%) 

10 (20%) 

2 (4%) 

• (50) .. 

1 (2%) 

(50) 

(48) 

(49) 

(50) 

(49) 

(50) 

(50) 

1 (2%) 

1 (2%) 

(12) 

(50) 

1 (2%) 

10 (20%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

(1) 

1 (lOO%) 

Cardiovascular System 

Heart (5o) (50) (5o) 

Schwannoma benign 1 (2%) 2 (4%) 
242 Dietary Restriction, NTP TR 460 

TABLE Ela 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) • 

Ad Libitum- Weight-Matched 

Fed Control Control 337.5 mg/kg 

2-Year Study (continued) 

Endocrine System 

Adrenal corlex (50) (5o) (50) 

Adrenal modulla (50) (50) (49) 

Pbeochromocytoma malignant I (2%) 

Pheochromocytoma complex 1 (2 %) 

Pheochromocytoma benign 12 (24%) 7 (14%) 7 (14%) 

Bilateral, pheochromocytoma benign 3 (6%) 2 (4%) l (2%) 

Islets, pancreatic (50) (50) (50) 

Adenoma 2 (4%) I (2%) 5 (10%) 

Carcinoma 1 (2%) 1 (2%) 

Pituitary gland (50) (48) (49) 

Ganglioneuroma 1 (2%) 

Pars distalis, adenoma 14 (28 %) 19 (4O%) 18 (37%) 

Thyroid gland (50) (50) (50) 

C-cell, adenoma 3 (6%) 6 (12%) 6 02%) 

C-cell, carcinoma 2 (4%) 1 (2%) 

Follicular cell, adenoma 4 (8%) 1 (2%) 

Follicular cell~ carcinoma I (2%) 

General Body System 

Tissue NOS (I) (I) 

Sarcoma. metastatic, intestine large, cecum 1 (100%) 

Genital System 

Epididymis 

Preputial gland 

(50) 

(50) 

(5O) 

(50) 

(5O) 

(50) 

Adenoma 1 (2%) 

Carcinoma 

Prostate . . . 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

,Interstitial cell, adenoma 

2 (4%) 

(50) 

(50) 

(50) 

44 (88%) 

1 (2%) 

(50) 

(50) 

(50) 

37 (74%) 

8 (16%) 

(50), 

(49) 

(50) 

43 (86%) 

2 (4%) 

Hematopoiefic Syst'em 

Bone marrow (50) (50) (50) 

Lymphnode (9) (15) (1o) 

Inguinal, fibrous histiocytoma, metastatic, 

skin 1 (10%) 

Mediastinai, sarcoma, metastatic,, intestine 

large, cecum 1 (7%) 

Lymph node, nmdibular (50) (49) (49) 

L.ymph node, mesenteric (50) (50) (49) 

Sarcoma, metastati~,:intestine large, cecur~ 1 (2%) 
=:~--~, - ::=~: ~ . - ~ ~ _ ~ ----:~.,.,~ ~,-~ _ =~-~__ ..... :: : z~: .: ~:= ..... ~ ~ - - ~__ , 

\ 

Dietar~'y Restriction, NT1P T~ ~6~ 2~3 

i 

i 

TABLE Ela 

S~n~raary o~ the ]Incidence o~ Neoplasms ~n Male Rats in the Dietary Restriction Study o~ Sa~icy~azosu~fapyrid~ne: 

Ad Libitum ~ee~ing ara~ Weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Hematopoietic System (continued) 

Spleen . 

Fibrosarcoma 

Leiomyosarcoma, metastatic, stomach, ! forestomach 

Sarcoma, metastatic, intestine large, cecum 

Thymus 

Thymoma benign 

Integumentary System 

Mammary gland 

Fibroadenoma 

Skin 

Basal cell adenoma 

Keratoacanthoma 

Squamous cell papilloma 

Trichoepi~elioma 

Subcutaneous tissue, fibroma 

Subcutaneous tissue, fibrosarcoma 

Subcutaneous tissue, fibrous histiocytoma 

Subcutaneous tissue, sehwannoma malignant 

Musculoskeletal System 

Bone 

Osteosareoma 

Skeletal muscle 

Fibrous histiocytoma 

Sarcoma, metastatic, intestine large, cecum 

Nervous System 

.Brain 

Astrocytoma malignam 

Granular cell tumor benign 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Osteosarcoma, metastatic, bone 

Sarcoma, metastatic, intestine large, cecum 

Nose 

Squamous cell carcinoma 

Ad Libitum- Weight-Matched 

Fed Control Control 

(50) (50) 

I (2%) 

1 (2%) 

1 (2%) 

(47) (50) 

1 (2%) 

(48) (50) 

3 (6%) 1 (2%) 

(50) (50) 

2 (4%) 

3 (6%) 2 (4%) 

1 (2%) 2 (4%) 

I (2%) 

2 (4%) 3 (6%) 

(2%) 

(50) (50) 

(1) (1) 

1 (100%) 

1 (100%) 

(50) (50) 

I (2%) 

1 (2%) 

(50) (50) 

2 (4%) 1 (2%) 

i (2%) 

(50) (50) 

1 (2%) 

337.5 mg/kg 

(50) 

(49) 

(49) 

3 (6%) 

(50) 

2 (4%) 

1 (2%) 

• 

3 (6%) 

1 (2%) 

I (2%) 

(50) 

I (2%) 

(i) 

(50) 

(50) 

1 (2%) 

(50) 

i 
244 Dietary Restriction, NTP TR 460 

TABLE Ela 

Summary of the Incidence of Neoplasms 

Ad Libitum Feeding and Weight-Matched 

2-Year Study (continued) 

Special Senses System 

Zymbal' s ghmd 

Carcinoma 

Urinary System 

Kidney 

Renal tubule, adenoma 

Urinary bladder 

Papilloma 

Papilloma, multiple 

Systemic Lesions 

Multiple organs c 

Leukemia mononuclear 

Mesothelioma malignant 

Neoplasm Summary 

Total animals with primary neoplasms d 

15-Month interim evaluation 

2-Year study 

Total primary neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total benign neoplasms 

15-Month interim evaluation 

2-Year study . 

Total animals with malignant neoplasms 

15-Month interim evaluation 

2-Year study 

Total malignant neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with metastatic neoplasms 

2-Year study 

Total metastatic neoplasms 

2-Year study 

in Male Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 337.5.mg/kg 

(1 ) (2) 

I (100%) 2 (100%) 

(50) (50) (50) 

1 (2%) 

(50) (50) (50) 

4 (8%) 

2 (4%) 

(50) (50) (5oi 

13 (26%) 10 (20%) 3 (6%) 

I (2%) 1 (2%) 3 (6%) 

9 10 10 

49 49 48 

12 13 10 

144 128 124 

9 10 10 

49 49 47 

10 13 10 

114 109 112 

2 

23 16 

2 

30 19 12 

1 5 

9 13 

Number of animals examined microscopically at the site and the number of animals with neoplasm 

Ten control animals were examined at 6 months for comparisons with a stop-exposure group that was not included.in the dietary restriction study .... 

Number of ~mimals with any tissue examined microscopically 

Primary neoplasms: all neoplasms except metastatic neoplasms 
:~: ~ : ~ @ ~ : - ; ~ r : ~ ~ _ _ ~ _ ~ ........ ~ ,:--. ............ ~ - ...... :~- :=~,---~,,~=:' . ~ ................... - ..... . __ 

Die~'y ]~es~ricti~n, NTIP T~ ~6~ ~45 

TA~L~ Elb 

S~m~ry o~ ~he ~nc~dence o~ Neoplasms ~n M~e ~ 

2-Yes ~d 30-Month R~tr~cted ~eed ~ot~o~ a 

~n ~he D~e~a~ R~r~ct~on S~udy o~ S~cy~osu~p~d~ne: 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle 

Control 337.5 mg/kg 

Vehicle 

Control 337.5 mg/kg 

Disposition Summary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Accidental deaths 

Moribund 

Natural deaths 

Survivors 

Died last week of study 

Terminal sacrifice 

Animals examined microscopically 

61 

10 

2 

13 

2 

34 

61 

60 

10 

2 

6 

3 

39 

60 

49 

4 

28 

7 

10 

49 

50 

2 

14 

10 

1 

23 

50 

, 

"i~ ? 

i -

i

[ 

i 

15-Month ~nterim Evaluation 

Endocrine System 

Pituitary gland 

Pars distalis, adenoma. 

(10) 

1 (10%) 

(1o) 

1 (1o%) 

Genital System 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

(10) 

6 (60%) 

3 (30%) 

(10) 

3 (30%) 

6 (60%) 

Integumentary System 

Skin 

Subcutaneous tissue, lipoma 

(10) (10) 

I (10%) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

Hematopoietic System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

,., 
246 Dietary Restriction, NTP TR 460 

"['ABLE Elb 

Summary of the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of Salicylazosuifapyridine: 

2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year and 30-Month Protocols 

Alimentary System 

Intestine large, colon 

Chotdorna, metastatic, bone 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Adenoma 

Liver 

Fibrous histiocytoma, metastatic, skin 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Hisfiocytic sarcoma 

Osteosarcoma, metastatic; bone 

Mesentery 

Osteosarcoma, metastatic, bone 

Pancreas 

Histi0cytic sarcoma, metastatic, spleen 

Osteosarcoma, metastatic, bone 

Acinar cell, adenoma 

Pharynx 

Squaruous cell papilloma 

Salivary glands 

Parotid gland, fil~rosarcoma 

Stomach, forestomach 

Squamous cell papilloma 

Stomach, glandular 

Tongue 

Squamous cell carcinoma 

Squamous cell papilloma. 

Cardiovascular System 

Heart 

Schwalmoma maligpant 

Endocrine System 

Adrenal cortex 

Adenoma 

Adrenal medulla 

, Pheochromocytoma malignant 

Pheochrom0cytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Adenoma " ., 

Carcinoma 

2-Year Restricted Feed 

Vehicle 

Control 337.5 mg/kg 

(51) (49) 

(51) (50) 

(50) (50) 

(51) (50) 

(50) (50) 

i (2%) 

(51) (5o) 

1 (2%) 1 (2%) 

(20) (7) 

(51) (50) 

5 (10%) 1 (2%) 

(1) 

I (100%) 

(51) (50) 

1 (2%) . 

(2) 

1 (5o%) 

1 (50%) 

(51) (50) 

1 (2%) 3 (6%) 

(51) (50) 

1 (2%) 

(31) (~0) 

2 (4%) 

5 (10%) 1 (2%) 

3 (6%) 

(51) (50) 

.. 1 (2%) 1 (2%) 

30-Month Restricted Feed 

. Vehicle 

Control 337.5 mg/kg 

(49) (47) 

1 (2%) 

(49) (49) 

(49) (49) 

(49) (48) 

(49) (49) 

(49) , (50) 

I (2%) I (2%) 

. 1 (2%) 

3 (6%) 

1 (2%) 

I (2%) 

(I 4) (10) 

1 (7%) 

(49) (48) 

1 (2%) 

1 (2%) 

2 (4%) 4 (8%) 

(1) 

1 (1oo%) : : 

(49) •. (49) 

I (2%) 

(49) (50) 

1 (2%) 

(49) (50) 

(r) 

I (1oo%) 

(49) (49) 

1 (2%) 

(49) (56i

1 (2%) 

(48) (50) 

1 (2%) 1 (2%) 

" 10 (21%) 6 (12%) 

4 (8%) 2 (4%) 

(49) (48) 

4 (8%) 4 (8%) 

5 (10%) 2 (4%) ,. 
= =~--- ~-~-=~.._=~__~.-.~__.~.~ : .... ~ ~ ..... ~.~ , ~:.,~_~_.~.~.~,,~. :~ ~_: ........................ 

D~e~ry l~estriction, N'r~ TI~ 46~ 247

TABLE Elb 

Summary of the ~[ncigence of Neoplasms in Ma~e ~s in the Dietary Restriction Stugy of S~[cy~s~f~pyr~c~e: I 

2-Year aug 30-Mionth ~estr~cted Feeg ]Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

~ Control 337.5 mg/kg Control 337.5 mg/kg 

I 

2-Year and 30-Month Protocols (continued) 

' 

i 

] 

.! 

I 

[ 

Endocrine System (continued) 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Bilateral, C-cell, carcinoma 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, adenoma 

(51) 

13 (25%) 

(50) 

4 (8%) 

(49) 

14 (29%) 

(50) 

5 (10%) 

1 (2%) 

(49) 

14 (29%) 

(49) 

6 (12%) 

1 (2%) 

1 (2%) 

(49) 

12 (24%) • 

(49) 

1 (2%) 

5 (10%) 

I (2%) 

1 (2%) 

~ ~ 

: 

i -

i 

, )! 

- i 

:[ 

::l General Body System 

Non° 

I 

I 

Genital System 

Epididymis 

Preputial gland 

Adenoma 

Carcinoma 

Prostate 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

(51) 

(51 ) 

3 (6%) 

(51) 

(51) 

(51) 

39 (76%) 

6 (12%) 

(50) 

(50) 

1 (2%) 

2 (4%) 

(50) 

(50) 

(50) 

39 (78%) 

6 (12%) 

(49) 

(49) 

2 (4%) 

4 (8%) 

(49) 

(49) 

(49) 

39 (80%) 

4 (8%) 

(49) 

(49) 

1 (2%) 

1 (2%) 

(49) 

(48) 

(49) 

37 (76%) 

5 (10%) 

Hematopoietic System 

Bone marrow 

Lymph node 

Deep cervical, carcinoma, metastatic, 

thyroid gland 

Mediastinal, fibrous histiocytoma, 

, metastatic, skin . 

Mediastinal, histioeytic sarcoma, metastatic, 

spleen 

Mediastinal, osteosarcoma, metastatic, bone 

Renal, fibrous histiocytoma, metastatic, skin 

Renal, histiocytic sarcoma, metastatic, 

spleen 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Fibroma 

Histiocytic sarcoma 

• Osteosarcoma, metastatic, bone 

Thymus 

(51) 

(1 l) 

(51) 

(51) 

(51) 

(47) 

(50) 

(3) 

(50) 

(50) 

(50) 

1 (2%) 

(48) 

(49) 

(20) 

1 (5%) 

(48) 

(49) 

(49) 

1 (2%) 

(44) 

(5O) 

(10) 

1 00%) 

1 (10%) 

i (10%) 

1 (10%) 

I 00%) 

(49) 

(48) 

(49) 

1 (2%) 

(4O) 
Dietary Restriction, NTP TR 460 

TABLE-Elb 

Summaryof the Incidence of Neoplasms in Male Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

2-Year arid 30-Month Protocols (continued) 

Integmnentary System 

Manm~aty gland 

Carcinoma 

(50) (49) (49). (47) 

1 (2%) 

Fibroadenoma 2 (4%) 2 (4%) : 2 (4%) 

Fibrous histiocytoma, metastatic, skin 1 (2%). 

Skin (51)' (50) (49) (50) 

Basal cell adenoma 1 (2%) l (2%) 

Basal cell carcinoma 1 (2%) 

Keratoacanthoma 

Squamous cell papilloma " ' 

Triehoepitheliomi~ 

! (~2%) 

2 (4%) 

2 (4%) 2 (4%) 

2 "(4%) 

(2%) 

(6%) 

(4%) 

Subcutaneous tissue, fibroid 2 (4%) 5 (10%) (10%) 

Subcutaneous tissue, fibrosarcoma 3 (6%) 

Subcutanbous tissu6, fibrous histiocytoma 1 (2%) 2 (4%) 

Subcutaneous tissue, .hemangiosarcoma I (2%) 

Subcutaneous tissue, schwannoma NOS (2%) 

• , 

Musculoskeletai System 

Bone (51) (50) (49) ,. (50) 

Chordoma l (2%) • .. 

Hamartoma i (2%) 

Osteosarcoma 1 (2%) 2 (4%) 1 (2%) 

Skeletal muscle 

Fibrous histiocytoma, metastatic, skin 

(2) (2) (2) 

1 (5o~o) 

Osteosareoma, metastatic, bone 1 (50%) 

Nervous System 

Brain (51) . (50) (49) (50). 

Oligodendroglioma malignant . _ .. 1 (2%) . 

.~ , 

R~piratory System • 

Lung. . (51) (50) (49) (49) 

Alveolar/bronchiolar adenoma 2 (4%) ~ (2%) 

Alveolar/bronchiolar carcinoma 2 (4%) 

Carcinoma, metastatic, thyroid gland 1 (2%) 

Fibrotr; histiocytoma, metastatic, skin 1 (2%) 1 (2%) 

Histiocytic sarcoma, metastatic, spleen 1 (2%) 

Trachea (51) (50) (49) (49) • . 

Carcinoma, metastatic, thyroid gland . b (2%) 

Special Senses System 

Zymbal's gland (1) • : 

., "(2) ' 

Carcinoma 1 (100%) - t(5o%) 
Dietary Restriction, NTP T~ 460 249 

i 

TABLE Ebb 

Summary of the ~ncidence of Neoplasms in Male Rats in the Dietary Restriction Study of Salicylazosu~fapyridine: 

2-a/ear and 30-Month Restricted ~eed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

I Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

I 

I 

:I. 2-~rear and 30-Month Protocols (continued) 

! Urinary System 

I KidneYLipoma (51) (50) 

l (2%) 

(49) (50) 

Histiocytic sarcoma, metastatic, spleen 1 (2%) 

Renal tubule, adonoma 1 (2%) 

Renal tubule, carcinoma 1 (2%) 

: { Urinary bladder (51) (50) (49) (49) 

Papilloma 1 (2%) 

I 

I 

I 

Systemic Lesions 

~ i Multiple organs b (51) (50) (49) (50) 

i Histiocyticsarcoma 1 (2%) 

Leukemia mononuclear 11 (22 %) 2 (4%) 24 (49%) 8 (16%) 

Mesothelioma malignant 3 (6 %) 1 (2%) 1 (2%) 4 (8%) i 

i 

Neoplasm Summary 

Total animals with primary neoplasms e 

15-Month interim evaluation 10 9 

2-Year and 30-month protocols 49 48 46 48 

Total primary neoplasms 

15-Month interim evaluation 10 11 

2-Year and 30-month protocols 116 86 152 119 

Total animals with benign neoplasms 

15-Month interim evaluation 10 9 

2-Year and 30-month protocols 49 48 

Total benign neoplasms 

15-Month interim evaluation 10 11 

2-Year and 30-month protocols .95 76 '104 92 

Total animals with malignant neoplasms 

2-Year and 30-month protocols 18 9 34 23 

• Total malignant neoplasms 

2-Year and 30-month protocols 21 10 48 26 

Total animals with metastatic neoplasms 

30-Month protocol 3 4 

Total metastatic neoplasms 

30-Month protocol 9 14 

Total animals with uncertain neoplasmsbenign or malignant 

30-Month protocol 

Total uncertain neoplasms 

.30-Month protocol • 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically ~ . 

e Primary neoplasms: all neoplasms except metastatic neoplasms 
250 Dietary Restriction, NTP TR 460 

'I'A~ E2a 

Statistical AnalysB of Primary Neoplasms in Male Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad L~bitum Feeding and Weight-Matched Controls Protocols 

337.5 mg/kg x 337.5 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

~drenal Medulla: Benign Pheochromocytoma 

Overall rate a 15/50 (30%) 8/49 (16%) 9/50 (18%) 8/49 (16%) 

Adjusted rate b 38.2 % 27.5 % 25.6 % 27.5 % 

Terminal rate c 11/35 (31%) 4/23 (17%) 6/31 (19%) 4/23 (17%) 

First incidence (days) 653 635 591 635 

Life table test d P=0.324N P-0.501 

Logisdc regression test d P=0.127N P=0.552N 

Fisher exact test d P = 0.084N P = 0.518N 

Adrenal Medulla: Benign, Complex, or Malignant Pheochromocytoma 

Overall rate 16/50 (32%) 8•49 (16%) 10/50 (20%) 8/49 06%) 

Adjusted rate 39.8 % 27.5% 28.6% 27.5% 

Termimd rate 11/35 (31%) 4/23 (17%) 7/31 (23%i 4/23 (17%) 

First incidence (days) 653 635 591 635 

Life table test ." P=0.265N P=0.586 

Logistic_ regregsion test P=0.087N P=0.451N 

Fisher 6xact test f P=0.056N P=0.416N 

Mammary Gland: Fibroadenoma 

Overall ~rate 3•50 (6%) 3/50 (6%) 1/50 (2%) 3/50 (6%) 

Adjusted rote 8.6%. II.4% 3.2% 11.4% 

Terminal rate 3/35 (9%) 2123 (9%) 1/31 (3%) 2/23 (9%) 

First incidence (days) 729 (T) 684 729 (T) 684 

Life table test P=0.480 P=0.219 

Logistic regression test P=0.577 P=0.270 

Fisher exact test ' P=0.661N P=0.309 .~. 

Pancreas: Adenoma • 

Overall rate 12/50 (24%) I0/50 (20%) 12/50 (24%) 10/50 (20%) 

Adjusted rate 31.4% 35.9% 33.0% 35.9% 

Terminal rate 9/35 (26%) 6•23 (26%) 8/31 (26%) 6/23 (26%) 

First incidence (days) 684 704 591 704 

Life table test P=0.394 P=0.543 

Logistic regression test P=0.590 P=0.468N 

Fisher exact test P=0.405N P=0.405N 

Panci'eas: Adenoma or Carcinoma 

Overall rate 13/50 (26%) 10/50 (20%) 12/50 (24%) I0/50 (2O%) 

Adjusted irate 34.1% 35.9% 33.0% 35.9% 

Terminal rate 10/35 (29%) 6/23 (26%) 8/31 (26%) 6/23 (26%) 

First incidence (days) 684 704 591 704 

Life table test P=0.469 P=0.543 

Logistic n;gre~sion test P=0.526N P=0.468N 

Fisher exact test P=0.318N P=0.405N 
~.... z;- =_ - ~ ; : 7 - 7 = ' 7 7 ~ ~ . . . . . . ~..... _ ........ ,.... ~:= ; : .-~.-~7:-:'~--'-= . . . . . . 

D~et~ry ]Restriction, NTP T~ ~60 251

TABLE, E2a 

StatLstica~ Ana~ys~ o~ Prhnary Neoplasms in Ma~e IRats in the Dietary ]Restriction Study oC Sa~cyllaz~s~apyrid~ne: 

Ad Libitum ]Feeding and Weight-Matched ControBs ]ProtocoLs (continued) 

} 337.5 mg/kg × 337.5 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Pancreatic Islets: Adenoma 

[ Overall rate 2/~0 (4%) 5/50 (10%) 1/50 (2%) 5/50 (10%) 

Adjusted rate 5.3% 18.3% 2.9% 18.3% 

Terminal rate 1/35 (3%) 3/23 (13%) 0/31 (0%) 3/23 (13%) I 

First incidence (days) 684 613 703 613 i Life table test P=0.113 P=0.067 

i , Logistic regression test P=O.190 . P=0.093 

Fisher exact test P=0.218 P=O.102 

Pancreatic ~slets: Adenoma or Cardnoma 

Overall rate 3/50 (6%) 5/50 (10%) 2•50 (4%) 5•50 (10%) 

Adjusted rate 8.1% 18.3% 6.0% 18.3% 

Terminal rate 2/35 (6%) 3/23 (13%) 1/31 (3%) 3•23 (13%) 

First incidence (days) 684 613 703 613 

Life table test P=0.198 P=0.143 

Logistic regression test P=0.314 P=0.195 

Fisher exact test P=0.357 P=0.218 

PRuitary Gland (Pars Distalis): Adenoma 

Overall rate 14/50 (28%) 18/49 (37%) 19148 (40%) 18/49 (37%) 

Adjusted rate 34.3% 50.5% 51.5% 50.5% 

Terminal rate 9/35 (26%) 7•22 (32%) 12/29 (41%) 7•22 (32%) 

First incidence (days) 473 428 521 428 

Life table test P=0.064 P=0.394 

Logistic regression test P=0.216 P=0.506N , 

Fisher exact test P=0.238 P=0.468N 

Preputial Gland: Adenoma or Carcinoma 

Overall rate 3/50 (6%) 0/50 (0 %) 0/50 (0%) 0/50 (0%) 

Adjusted rate 7.7% 0.0% 

Terminal rate 1/35 (3%) 0/23 (0%) 

e First incidence (days) 684 _ 

Life table test P=0.185N 

Logistic regression test P=0.133N 

Fisher exact test P=0.121N 

Skin: Keratoacanthoma 

Overall rate 3/50 (6%) 2/50 (4%) 2/5,0 (4%) 2•50 (4%) 

Adjusted rate 8,6% 6,8% 

Terminal rate 3•35 (9%) 0/23 (0%) 

First incidence (days) 729 (T) 687 

Life table test P=0.655N 

Logistic regression test P=0,569N 

Fisher exact test P=0.500N 

J 

[ -: 

i 
252 ' Dietary Restriction; NTP TR 460 

/ 

'I'A~i~ E2a 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) " . . . . . . . 

337.5 mg/kg x 337.5 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Skin: Squamous Cell Papilloma or Keratoacanthoma 

Overal~ rate 4/50 (8%) 3/50 (6%) 3/50 (6%) 

Adjusted rate 11.4% 9.7 % 9.3 % 

Terminal rate ' 4/35 (I1%)' 0/23 (0%) " 2/31 (6%) 

First incidence (days) 729 (T) 687 709 

Life table test P=0.627 

Logistic regression test P=0.574N 

Fisher exact test P=0.500N 

Squamous Cell Papilloma, Keratoacanthoma, Trichoepithefioma, or Basal Cell Adenoma 

Overall rate 6/50 (12%) 3/50 (6%) 3/50 (6%) 

Adjusted rate 16.5% 9.7% 9.3% 

Terminal rate ' 5/35 (14%) 0/23 (0%) 2/31 (6%) 

First incidence (days) 717 687 709 

Life tabIe test P=0.440N 

Logistic regression test P=0.315N 

•Fisher..e~act test P=0.243N _, 

Slain (Subcutaneous Tissue): Fibroma 

Overall rate 2/50 (4 %) ' 3/50 (6%) 3/50 (6%) 

Adjusted rate 5.7% 12.5% 8.0% 

Terrnina~ rate 2/35 (6%) 2/23 (3 %) 1/31 (3%) 

First incidence (days) 729 (T) 726 620 

Life table test P=0.316 

Logistic regression test P=0.332 

Fisher exact test P=0.500 

Skin (Subcutaneous Tissue): Fibroma, Fibrous Histiocytoma, or Fibrosarcoma" . . 

Overall rate ' 2/50 (4%) 5/50 (10%) 3/50 (6%) 

Adjusted rate 5.7% 17.4% • 8.0% 

Terminal rate 2/35 (6 %) 2/23 (9%) 1/31 (3%) 

First incidence (days) 729 (T) 645 620 

Life table test P=0.108 

Logistic regression test P =0.172 

Fisher exact test P=0.218 

,. ',.:,. '.=~" 

Testes: Aden0ma 

Overall rate 45/50 (90%) 45/50 (90%) 45/50 (90%) 

Adjusted rate 97.8% 100.0% 97.8%

Terminal rate 34/35 (97%) 23/23 (100%) 30/31 (97%) 

First incidence (days) 477 473 458 

Life table test P=0.009 

Logistic regres.sion test P=0.405 

Fisher exact test P=0.630N 

~ / 

3/50 (6%) 

9.7% 

0/23 (0%) 

687 

P=0.556 

P=0.633 

P=0.661N 

3/50 (6%) 

9.7% 

0/23 (0%) 

687. 

P=0.556 

P=0.633 

P=0;661N 

3/50 (6%). 

12.5% -

2/23 (9%) .... 

726 " 

P=0.550 

P=0.642 • 

P=0.661N " 

5/50 (lo%y ~. - 

17.4% ...... 

2/23 (9%) ' 

645 " 

P=0.266 

P=0,338 .. .~ 

P=0,357 

45/50 (90%) 

100.0%' " ,'

23/23 (100%) " 

473 '. '. 

P=0.080 

-P=0.467 " ' - - " 

P=0.630N ' 
~ - - ~ . ~ _ _ ~ ................. ~,::-- .--~----~-~ ._ ~ ~---~-----~- ....... 

[ 

Die~'y Restriction, NTP TR ~0 253 

T~L~ E2a 

S~L~tic~fl An~flys~s of Pr~nm'y NeopLasms in I~hfle RaGs in the Die~'y ]Restriction Study o~ Sa~icy~zosuffapyrid~ne: 

Ad LibL~tm Feeding and Weight-Matched Controls Pr'otoeo~s (continued) 

I 

I 

1 

[ 

I 

Thyroid Gland (C-cell): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Adenoma 

Ad Libitum, 

Fed Control 

3/50 (6%) 

8.6% 

3•35 (9%) 

729 (T) 

337.5 mg/kg x 

Ad LibitumFed Control 

6/50 (12%) 

22.9% 

4/23 (17%) 

696 

P=0.093 

P=0,145 

P=0.243 

337.5 mg/kg x 

Weight-Matched Weight-Matched 

Control Control 

6/50 (12%) 6/50 (12%) 

18,2% 22.9% 

5/31 (16%) 4/23 (17%) 

646 696 

P=0,438 

P=0.556 

P=0.620N 

l 

i 

Thyroid Gland (C-cell): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Adenoma or Carcinoma 

5/50 (10%) 

14.3% 

5/35 (14%) 

729 (T) 

7/50 (14%) 

24.9% 

4•23 (17%) 

653 

P=0.162 

P=0.278 

P=0.380 

6/50 (12%) 

18.2% 

5/31 (16%) 

646 

7/50 (14%) 

24.9% 

4/23 (17%) 

653 

P=0.328 

P=0.446 

P=0.500 

Thyroid Gland (Follicular Cell): Adenoma 

Overall rate 4/50 (8%) I150 (2%) 0/50 (0%) 1/5o (2%) 

Adjusted rate 

Terminal rate 

11.4% 

4/35 (11%) 

4.3% 

I123 (4%) 

First incidence (days) 729 (T) 729 (T) 

Life table test P=0.324N 

Logistic regression test P=0.324N 

Fisher exact test P=0.181N 

Thyroid Gland (Follicular Cell): Adenoma or Carcinoma • 

Overall rate 5/50 (10%) 1/50 (2%) 0/50 (0%) 1/50 (2%) 

Adjusted rate 14.3% 4.3% 

Terminal rate 5/35 (14%) 1/23 (4%) 

First incidence (days) 729 (T) 729 (T) 

Life table test P=0.221N 

Logistic regression test P=0.221N 

Fisher exact test P=0.102N 

Urinary Bladder: Papilloma 

Overall rate .. 0/50 (0%) 6/50 (12%) 0/50 (0%) 6i50 (12%) 

Adjusted rate 0.0% 22.1% 0.0% 22.1% 

Terminal rate ., 0/35 (0%) 3/23 (13%) 0/31 (0%) 3/23 (13%) 

First incidence (days) 653 - - 653 

Life table test P=0.006 P=0.009 

Logistic regression test P=0.011 P=0.013 

Fisher exact test P=0.013 P=0.013 
254 

--7 

Dietary Restriction, NTP TR 460 

TABLE E2a 

Statistiicall Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study of Salicylaz0sulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

337.5 mg/kg × 337.5 mg/kg × 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

All Organs:: Mononuclear Cell Leukemia 

Overall rate 13/50 (26%) 3/50 (6%) ~ 10/50 (20%) 3/50 (6%) 

Adjusted rate 32,0% 6.8% 26.2% 6.8% 

Terminal rate 8/35 (23%) 0•23 (0%) 5/31 (16%) 0/23 (0%) 

First incidence (days) 477 428 526 428 

Life table test P=0,040N P=0.076N 

Logistic regression test P=0.007N P=0.033N 

Fisher exact test , ' P=0.006N P=0.036N 

All Organs: Malignant Mesotheli0ma ' 

Overall rate 1/50 (2%) 3/50 (6%) 1/50 (2%) 3/50 (6%) 

Adjusted rate 

Terminal rate . • 

,.~ 

~ ', 

2.9% 

1/35 (3%) 

10.3% 

1/23 (4%) " 

3.2% 

1731 (3%) 

.10.3% 

1/23 (4%) 

First incidence (days) 

Life table test • 

729 (T) 585 

. P=0.207 

729 (T) 585 

P=0.236 

Logistic regression test 

Fisher exact test 

P=0.292 

P=0.309 

P=0.295 

P=0.309 

. 

• , 

All.Organs:, Benign Neoplasms 

Overall rate 

: 

49/50 (98%) 47/50 (94%) 49/50 (98%) . '" 47/~0 (94%) 

Adjusted rate 

Terminal rate 

100.0% 

35/35 (100%) 

100.0% 

23/23 (100%) 

100.0% 

31/31 (100%) 

100.0% 

23/23 (100%) 

First incidence (days) 

Life table test 

473 428 

P=0,025 

458 428 

P=0.144 

Logistic regression test 

Fisher exact test 

P=0.998N 

P=0.309N 

P=0,970N 

P~0.309N 

All Organs: 

Overall rate 

Malignant Neoplasms 

23/50 (46%) 9/50 (18%) 16/50 (32%) 9/50 (18%) 

Adjusted rate 

Terminal rate 

49.9% 

12/35 (34%) 

22.8% 

1/23 (4%) 

39.5% 

8/31 (26%) 

22.8% 

1/23 (4%) 

First incidence (days) 

Life table test 

477 428 

P=0.049N 

484 428 

P=0.198N 

Logistic regression test 

Fisher exact test 

P=0,002N 

P=0.002N 

P=0.072N 

P=0.083N 
::- :-~--~ ~:~: - --~-~S-S~-:~-~ ~_.~-~ i m l i ~ m l a ~ . =. = = ~ ~~ - ~ _ _ _ . . . . . . . . . ~ ....... , ~ .......... ~ - ~ ......................... 

i 

D~e~ry ~estricfion~ ~ TII~ ~60 2~5 

TA~Lg E2~ 

S~s~c~ A~llys~s o~ ]~'h~ry Neoplasms ~n 1~I~e 1~ ~n ehe D~eL~ry ~e~c~on 

Ad Libi~m Fee~ng ~nd We~gh~-I~iI~ched Con~ro~s i~ro~ocoLs (continued) 

S~dy o~ S~cyll~~yr~ne: 

I 

Ad LibitumFed Control 

337.5 mg/kg × 

Ad LibitumFed Control 

Weight-Matched 

Control 

337.5 mg/kg × 

Weight-Matched 

Control 

I : 

All Organs: ~enign or Malignant Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

49/50 (98%) 

100.0% 

35/35 (100%) 

473 

48150 (96%) 

100.0% 

23/23 (100%) 

428 

P=0.018 

-f 

P=0.500N 

49/50 (98%) 

100.0% 

31/31 (100%) 

458 

48/50 (96%) 

100.0% 

23/23 (100%) 

428 

P=0.113 

-

P=0.500N 

I 

i 

?1 

i 

i 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, 

pancreas, pancreatic islets, pituitary gland; preputial gland, testes, thyroid gland, and urinary bladder; for other tissues, denominator is number of animals 

necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

c Observed incidence at terminal kill 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the ad libitum-fedor weight-matched controls and the 

dosed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in the 

dosed group is indicated by N. 

e Not applicable; no neoplasms in animal group 

f Value of statistic cannot be computed. 
256 Dietary Restriction, NTP TR460 

TABLE: E2b 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study 

of Salicylazosuifapyridine: 2-Year and 30-Month Restricted Feed Protocols 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg ... 

Adrenal Medulla: Benign Pheochromocytoma 

Overall rate a 

Adjusted rate b " 

Terminal rate c 

First incidence (days) 

Life table test d 

,. 8/51 (16%) 

22.2% 

6/34 (18%) 

700 

1/50 (2%) 

3.4% 

1/39 (3 %) 

731 (T) 

P=0.011N 

' 14/48 (29%) 

66.7% 

4/10 (40%) 

752 

8/50 (16%) 

31.1% 

7/24 (29 %) 

812 

P=0.002N 

Logistic regression test d 

Fisher exact test d 

' P=0.014N 

P=0.017N 

P=0.011N 

P=0.093N ., 

Adrenal Medulla: Benign or Malignant Pheochromocytoma 

Overall rate 9/51 (18%) 

Adjusted rate 33.3 % 

Terminal rate 7/34 (21%) 

First incidence (days) ~ 700 

Life table test 

1/50 (2%) 

3.4% 

1/39 (3%) 

731 (T) 

P=0.006N 

15/48 (31%) 

72.3% 

5/10 (50%) 

752 

8/50 (16%) 

31.1% 

7/24 (29 %) 

812 

P<0.001N 

., 

Logistic regression test 

Fisher exact test " 

P=0,007N 

P=0.009N 

P=0.005N 

P=0.061N 

Heart: Malignant Schwannoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

1/51 (2%) 

2.9% 

1/34 (3 %) 

730 (T) 

., 

3)50 (6%) 

10.3% 

3/39 (8%) 

731 (T) 

P=0.355 

., 

,, ; ~ . 

1/49 (2%) 

' . 

0/49. (0 % ) 

• . 

Logistic regression test 

Fisher exact test 

P=0.355 

P=0.301 

Liver: Hepatocel!ular Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regressioh test " 

Fisher exact test 

:, 

-: 

• 

• 

,. 

1/51 (2%) 1)50 (2%) 3/49 (6%) 

16.9% 

0/10 (0%) 

816 

0/50 (0%) 

0.0% • 

0/24 (0%) 

P=0.047N 

P=0.082N , 

P=0.117N 

Liver: Hepatocellu!ar Adenoma or Carcinoma 

Overall rate : 1/51 (2%) .~ 

Adjusted rate • ~. 

Terminal rate 

First incidence (day~) 

Life table test -~ 

Logistic regression ~e,st ~-. 

Fisher exact test 

'1/50 (2%) 3/49 (6%) 

16.9% 

0/10 (0%) 

816 

.~ • . , 

1/50 (2%) 

3.3% 

0/24 (0 K) 

854 

P=0.146N 

P=0.232N 

P=0.301N 

;. 
I 

257 

. 7 ~ - - ::::: --=~ ~'~ i~----'~:=--~ ,~_ ~ ~ . . . . . . . . . . . . ~--'~'!--~ ~ ....... : ........ ~ .... =:~: ; '~:r~'-:--- ~ 

Dietary Restriction, NT~ TR ~6~ 

TABLE E2b 

Stafistica~ Analysis of Primary Neoplasnas in Male ]Rats in the Dietary ~estr~ction Study 

of Saiicyiazosu~fapyridine: 2-Year and 30-Month Restricted Feed ~?rotoeo~s (continued) 

2-Year Restricted Feed " 30-Month Restricted Seed 

Lung: Alveolar/bronchiolar 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test. 

Vehicle Vehicle 

Control 337.5 mg/kg Control 

Adenoma or Carcinoma 

2/51 (4%) 

Mammary Gland: Sibroadenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Oral Cavity (Tongue and Pharynx): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life tabl~e test 

Logistic"regression test 

Fisher exact test 

Pancreas: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression tes~ 

Fisher exact test 

Pancreatic Islets: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test • 

Fisher exact test " ' 

. 2/51 (4%) 

,., 

Squamous Cell Papilloma or 

3/51 (6%) 

6.6% 

0/34 (0%) 

627 

5/51 (10%) 

23.5% 

3134 (9%) 

667 

I151 (2%) 

0/50 (0%) 3/49 (6%i 

21.8% 

1/10 (10%) 

869 

0/50 (0%) 2/49 (4%) 

13.8% 

1/10 (10%) 

816 

Squamous Cell Cardnoma 

0/50 (0%) 1/49 (2%) 

0.0% 

0/39 (0%) 

P=0.132N 

P=0.102N 

P=0.125N 

1/50 (2%) 2/49 (4%) 

2.4% 20.0% 

0•39 (0%) 2/10 (20%) 

674 910 (T) 

P=0.088N 

P=0.111N 

P=0.107N 

1/50 (2%) .4•49 (8%) 

19.4% 

1/10 (10%) 

660 

337.5 mg/kg 

0/49 (0%) 

0.0% 

0•24 (0%) 

P=0.035N 

P=0.049N 

P=0.121N 

3/50 (6%) 

12.5% 

3/24 (13%) 

910 (T) 

P=0,533N 

P=0.669N 

P=0.510 

1/50 (2%) 

,~/48 (8%) 

14.9% 

3/24 (13% 

795 

P=0.645N 

P=0.535 

P=O.a2b 

' 4/48i8%) ' 

16.7% 

4/24 (17%) 

910 (T) 

P=0.287N 

P=0.569N 

P=0.631 
258 Dietary Restriction, NTP TR 460 

TABLEE2b 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study 

of Salicy]lazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

Pancreatic Islets: Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pancreatic Islets: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test • 

Fisher exact test 

Preputial Gland: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate ) " ̧ 

First incidence (days) . 

Life table test 

Logistic regression test 

Fisher exact te:;t 

Preputial Gland: Carcinoma" 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Vehicle 

Control 

0151 (0%) 

1/51 (2%) 

13/51(25%) 

31.1% 

7/34 (21%) 

520 

3/51 (6%) 

8.8% 

3/34 (9%) ' 

730 (T) 

? 

0/51 (0%) 

337.5 mg/kg 

0/50 (0 %) 

1/50 (2%) 

14/49 (29%) 

40.2% 

11/38 (29%) 

634 

P=0.580N 

P=0.428 

P=0.451 

1/50 (2%) 

3.4% 

1/39 (3%) 

731 (T) • 

P=0.257N 

P=0.257N 

P=0.316N 

2/50 (4%) 

30-Month Restricted Feed 

Vehicle • 

Control 

5/49 (10%) 

23.2% 

O/lO (0%) 

763 

8/49 (16%) 

35.0% 

1/10(10%) 

660 

14/49 (29%) 

51.4% 

2/10 (20%) 

556 

2/49 (4%) 

4/49 (8%) 

23.6% 

: 1/I0 (10%) 

571 

337.5 mg/kg 

2/48 (4%) 

6.8% 

1/24 (4%) 

812 

P=0.076N 

P=0.177N 

P=0.226N . 

6/48 (13%) 

23.0% 

5/24 (21%) 

812 

P=0.079N 

P=0.294N 

P=O.403N 

12/49 (24%) 

38.8% 

7/23 (30%). 

573 

P=0.081N 

P=0.390N 

P=0.410N 

~ , 

1/49 (2%) 

1/49 (2%) 

4.2% 

1/24 (4%) 

910 (T) 

P=0.053N 

P=0A52N 

P=0.181N, 
.~- ~= :~ ~= -- "~_~7:~7--.~-~:~._~. ~ . ~ . -- : ~ ..... ~.,_~,~.,~.~-- . 2 --~-.-- . --_..~7-~;~;~ ...... ....... 

:[ 

I 

I 

: I 

I 

Dietary Sestricfion, N'rP TR 460 

TABLE E2b 

StatBtical Analysis o~ Primary Neoplasms in Male Rats in the Dietary Restriction,Study 

off Sa~icylazosulfapyridine: 2-Year and 30-Month Restricted Feed ]?rotocoh (continued) 

2oYear Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

Preputial Gland: Adenoma or Carcinoma 

Overall rate 3/51 (6%) 3/50 (6%) 6/49 (12%) 2/49 (4%) 

Adjusted rate 8.8% 10.3% 36.3% 8.3% 

Terminal rate 3•34 (9%) 3/39 (8%) 1/10 (10%) 2/24 (8%) 

First incidence (days) 730 (T) 731 (T) 571 910 (T) 

Life table test P=0.599N P=0.018N 

Logistic regression test P=0.599N Pffi0.078N 

Fisher exact test P=0.652 P=0.134N 

Skin: Squamous Cell Papilloma 

Overall, rate 0/51 (0%) 0/50 (0%) 2/49 (4%) 3/50 (6%) 

Adjusted rate 9.0% 12.5% 

Terminal rate O/lO (0%) 3/24 (13%) 

First incidence (days) 816 910 (T) 

Life table test P=0.567N 

Logistic regression test P=0.649 

Fisher exact test P=0.510 

Skin: Squamous Cell Papilloma or Keratoacanthoma 

Overall rate 1/51 (2%) 2/50 (4%) 4/49 (_8%) 4/50 (8%) 

Adjusted rate 27.2% 16.7% 

Terminal rate ' 2/10 (20%) 4•24 (17%) 

First incidence (days) 816 910 (T) 

Life table test P=0.223N 

Logistic regression test P=0.391N 

Fisher exact test P=0.631N 

Skin: Trichoepithelioma, Basal Cell Adenoma, or Basal Cell Carcinoma 

Overall rate 3/51 (6%) 1/50 (2%) 1/49 (2%) 2/50 (4%) 

Adjusted rate 19.3 % 3.4 % 7.7% 8.0% 

Terminal rate 3/34 (9%) 1/39 (3%) 0/10 (0%) 1/24 (4%) 

First incidence (days) 730 (T) 731 (T) 897 889 

Life table test P=0.257N P=0.726N 

Logistic regression test P=0.257N • P=0.691 

Fisher exact test P=0.316N P=0.508 

Skin: Basal Cell Papilloma, Keratocanthoma, Trichoepithelioma, Basal Cell Adenoma, or Basal Cell Carcinoma 

Overall rate 4/51 (8%) 3/50 (6%) 5/49 (10%) 6/50 (12%) 

Adjusted rate 32.8% 10.3% 32.8% 24.0% 

Terminal rate 4•34 (12%) 3/39 (8%) 2/10 (20%) 5/24 (21%) 

First incidence (days) 730 (T) 731 (T) 816 889 

Life table test P=0.425N P=0.256N 

Logistic regression test P=0.425N P=0.435N 

Fisher exact test P=0.511N P=0.514 

259 

i 

i 

J 
-- 

Dietary Restriction, NTP TR 46~ 

TABLEE2b 

Statistical Analysis: of Primary Neoplasms in Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

S~kin (Subcutaneous Tissue): Fibroma 

Overall ~ate 

Adjusted rate. ~ 

Terminal rate 

First incidence (days) ~ 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): Fibrosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Testes: Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

2-Year Restricted Feed 

Vehicle 

Control 337.5 mg/kg 

2151 (4%) 0150 (0%) 

0151 (0%) 0/50 (0%) 

Fibrous Histiocytoma or Fibrosarcoma 

0/51 (0%) 0/50 (0%) 

Fibroma, Fibrous Histiocytoma, or Fibrosarcoma 

2/51 (4%) 0/50 (0%) 

"45/51 (88 %) 45/50 (90 %) 

•100.0% 97.8% 

32/34 (94 %) 38/39 (97 %) 

520 511 

P=0.218N 

P=0.255 

' P=0.514 

30-Month Restricted Feed 

Vehicle 

Control 

.. , 

5/49 (10%) 

33.3% 

2/10 (20%) 

690 

3/49 (6%) 

18.8% 

1/10 (10%) 

816 

4/49 (8%) 

27.8% 

2/10 (20%) 

816 

8/49(16%) 

52.1% 

4/10 (40%) 

690 

43/49 (88%) 

97.7% 

9/i0.(90%) 

445 

337.5 mg/kg 

5/50 (10%) 

16.4% 

2/24 (8.%) 

737 

P=0.254N 

P=0.512N 

P=0.617N 

O/50 (0%) 

0,0% 

0/24 (0%) 

P=0.042N 

P=0.076N 

P=0.117N 

5.8% 

0/24 (0%) 

199 

~P=Q.134N 

P=0.334N 

P=0.329N 

7•50 (14%) 

21.2% 

~2/24 (8%) 

199 

P = 0.116N 

P=0.441N 

P=0.483N 

42/49 (86 %) 

ioo.0% 

•24/24 (100%) 

430 

P=0.001N 

P=0~431N 

P=0.500N 

.. 
:S~: :? =%=-'-~--~ ~ - - - - ........................ ~:--r-_--~-~- ~ ~:7~'-~-~ ~--~-. ~ ~ . ..................... ,, ,., ..... 

i 

Dietary Restriction, NTP TR 4~ 26~ 

! 

i 

TABLE E2b • 

Statistical Ana~ysls o~ ?rimary Neoplasms in Ma~e Rats ira the Dietary Restriction Study 

o~ Sa~icylazosu~ffapyridine: 2-Year and 30-Month Restricted ~eed ]Protocols (continued) 

J 

j = 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle 

Control 337.5 mg/kg 

Vehicle 

Control 337.5 mg/kg 

I 

Thyroid Gland (C-cell): Adenoma 

Overall rate 4/50 (8%) 

Adjusted rate 32.8% 

Terminal rate 4/34 (12%) I 

First incidence (days) 730 (T) 

Life table test 

Logistic regression test 

Fisher exact test " 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 4•50 (8%) 

Adjusted rate 32.8% 

Terminal rate 4/34 (12%) 

First incidence (days) 730 (T) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Mononuclear Cell Leukemia 

Overall rate 11/51 (22%) 

Adjusted rate 43.4% 

Terminal rate 3/34 (9%) 

First incidence (days) 520 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Malignant Mesothelioma 

Overall rate 3/51 (6%) 

Adjusted rate 8.2% 

Ter, minal rate 2/34 (6%) 

First incidence (days) 682 

Life table test 

Logistic regression test 

Fisher. exact test 

All Organs: Benign Neoplasms 

Overall rate 49/51 (96%) 

Adjusted rate 100.0% 

Terminal rate ; .. 34/34 (100%) 

First incidence (days) 520 

Life table test 

Logistic regression test 

Fisher exact test 

5/50 (10%) 

14.8% 

4/39 (10%) 

685 

P=0.579 

P=0.525 

P=0.500 

6/50 (12%) 

18.1% 

5/39 (13%) 

685 

P=O.452 

P=O.397 

P=O.37o 

2•50 (4%) 

6.9% 

2•39 (5%) 

731 (T) 

P=0.009N 

P=0.010N 

P=0.008N 

1/50 (2%) 

3.4% 

1/39 (3%) 

731 (T) 

P=0.269N 

P=0.309N 

P=0.316N 

48/50 (96%) 

100.0% 

39•39 (100%) 

210 

P=0.157N 

P=0.112 

P=0.684N 

6/49 (12%) 

28.3% 

1/10 (10%) 

594 

7/49 (14%) 

31.8% 

1/10 (10%) 

594 

24/49 (49%) 

71.3% 

3/10 (30%) 

556 

1/49 (2%) 

4.2% 

0/10 (0%) 

816 

46/49 (94%) 

100.0% 

10/10 (100%) 

445 

5/49 (10%) 

13.2% 

1/24 (4%) I 

571 

P=0.281N 

P=0.503N 

P=0.500N 

6•49 (12%) 

17.0% 

2/24 (8%) i 

571 

P=0.235N i 

P=0.491N 

P=0.500N 

8/50 (16%) 

23.8% 

3/24 (13 %) 

430 

P<0.001N 

P<0.001N 

P<0.001N 

4•50 (8%) 

12.3% 

1/24 (4%) 

737 

P=0.343 

P=0.204 i 

i 

P=0.187 

i 

46/50 (92 %) 

100.0% 

24/24 (100%) 

430 

P=0.002N 

P=0.654N 

P=0.511N 
262 ~ Dietary Restriction, NTP TR460 

TABLE E2b 

Statistical Analysis of Primary Neoplasms in Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

All Organs: Malignant Neoplasms 

Overall rate 18/51 (35%) 9/50 (18%) 34/49 (69%) 23/50 (46%) 

Adjusted rate 61.8 % 29.3 % 85.3 % 56.4% 

Terminal rate 7/34 (21%) 8/39 (21%) 5/10 (50%) 8/24 (33%) 

First incidence (days) 520 674 556 199 

Life table test P =0.03 IN P < 0.00IN 

Logistic regression test P=0.045N P=0.012N 

Fisher exact test P=0.040N P=0.015N 

All Organs: Benign or Malignant Neoplasms 

Overal| rate . . . . 49/51 (96%) 48/50 (96%) 46/49 (94%) 48/50 (96%) 

Adjusted rate 100.0% 100.0% 100.0% 100.0% 

Terminal rate 34/34 (100%) 39/39 (100%) 10/10 (100%) 24/24 (100%) 

First incidence (days) 520 210 445 199 . 

Life table test P=0.157N P=0.004N 

Logistic regression test : 'P=0.112 P=0.206 -

Fisher exact test P=0.684N P=0.490 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/numberof animals examined. Denominator is number of animals examined microscopically for adrenal gland, heart, 

liyer, lung, pancreas, pancreatic islets, pituitary gland, preputial gland, testes, and thyroid gland; for other tissues, denominator is number of animals 

necropsied~ 

b Kaplan-Meier estimated neoplasm incidence at the end of the ~tudy after adjustment for intercurrent mortality 

c Observed incidence at terminal kill • 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the controls and that dosed group. The life table test 

regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesio~ts as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in a dosed group is indicated by N. 

• Not applicable; no neoplasms in animal group 

 ̧.," 
I

I : 

• D~e~ry 1~est~ct~on, ~ T~ ~360 263 

i ~ 

i

i ?z ! ~ 

i ~: 

TAII~L~ ].E3a 

I ~ ~ 

_ :. 

S~m'y o~ the ]Incidence o~ Nouneop~mfic Lesions ~n Mate ~a~ ~u the D~et~'y ~estr~ct~on Study 

o~ S~cy~os~apyr~d~ne: Ad Libigum ~eed~ng and We~ght-IVi[atched ControLs ~?rotocoLs a i 

~ 

Ad LibitumFed Control 

Weight-Matched 

Control 337.5 mg/kg 

I 

I 

I 

I 

! 

~ 

~ 

= 

DLsposition Smnnmry 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Accidental deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

70 b 

I0 

1 

13 

1 

35 

60 

10 

2 

16 

1 

31 

60 

I0 

2 

15 

10 

'23 

Animals examined microscopically 60 60 60 

£ 5-M~ongh ~nger~m E~,aluation 

Alhnentary System 

Intestine large, colon 

parasite metazoan 

Intestine large, rectum 

Parasite metazoan 

Liver 

Basophilic focus 

Clear cellfocus 

Eosinophilic focus 

Granuloma 

Hepatodiaphragmatic nodule 

Inflammation, chronic 

Mixed cell focus 

Bile duct, hyperplasia 

Hepatocyte, vacuolization cytoplasmic 

Lobules, necrosis 

Mesentery 

Fat, hemorrhage 

Fat, metaplasia, osseous 

Fat, necrosis 

Pancreas 

Atrophy 

Acinar cell, hyperplasia, focal 

(10) 

(10) 

1 (10%) 

(10) 

2 (20%) 

5 (50%) 

2 (20%) 

1 (10%) 

2 (20%) 

8 (80%) 

8 (80%) 

(2) 

1 (50%) 

1 (50%) 

(10) 

3 (30%) 

2 (20%) 

(10) 

1 (10%) 

(10) 

2 (20%) 

(10) 

3 (30%) 

2 (20%) 

1 (10%) 

3 (30%) 

1 (10%) 

2 (20%) 

5 (50%) 

5 (50%) 

(3) 

1 (33%) 

2 (67%) 

(10) 

5 (50%) 

3 (30%) 

(lO) 

1 

(io) 

(io) 

3 

1 

(2) 

(10%) 

(30.%) 

(lO%) 

(1o%) 

(30%) 

(3o%) 

(10%) 

2 (10o%) 

(lO) 

5 (5o%) 

3 (30%) 

i ~ 

i 

J 

i 

i 

!

i 

i 

I 

Cardiovascular System 

Heart 

Cardiomyopathy 

Pericardium, fibrosis 

(10) 

6 (60%) 

(10) 

3 (30%) 

(10) 

3 (30%) 

I (10%) 

a 

b 

Number of animals examined microscopically at the site and the number of animals with lesion 

Ten control animals were examined at 6 months for comparisons with a stop-exposure group that was not included in the dietary restriction study. 
264 Dietary Restriction, NTP TR 460 

TABLE E3a 

Summary of the Incidence of Nonneoplastic Eesions in Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols (conti.nued) 

Ad Libitum- Weight-Matched 

Fed Control Control 337.5 mg/kg 

15-Month Interim Evaluation icontinued) 

Endocrine System 

Adrenal cortex (lO) (lO) (lO) 

Accessory adrenal cortical nodule 1 (10%) 2 (20%) 5 (50%) 

Degeneration, fatty t 00%) 

Hyperplasia, focal : 1 (10%) 

Islets, pancreatic (10) (10) (10) 

Hyperplasia 1 (10%) 

Pituitary gland (1o) (tO) (tO) 

Pars distalis, angiectasis 1 (1o%) 

Pars distalis, cyst 1 (lO%) 1 (10%) (40%) 

Pars distalis, hyperplasia, focai 3 (30%)  ̧,1 (10%) 

Thyroid gland (lO) (to) (I0) 

C-cell, hyperplasia I (1o%) 

Follicle, ~yst t (10%) 1 (10%) I (10%) 

Follicular cell, hypertrophy 2 (20%) 

• , 

Genital System 

Epididymis (10) (10) (10) 

' Atypia cellular 1 (10%) 3 (30%) 

PrepUtial gland 

Inflanun#tion, chronic 

il0) 

8 (80%) 

(to) 

8 (80%) 

(to) 

6 

• 

(60%) 

Inflanun,~tion, suppurative 12 (20%) 2 (20%) 

Prostate (lO) (10) (I0) 

Corpora amylacea 2 (20%) t (10%) 2 (20%) ,, 

Fxlema 1 (10%) 

Inflammation, suppurative 

Testes 

• 6 

(lo) 

(60%) - 5 (50%) •

(lO) 

3 

(1o) 

(30%) 

Interstitial cell, hype~plasia 4 (40%) 4 (4O%) 1. 00%) 

Hematopoietic System 

Lymph node (i) 

Mediastinal, hemorrhage 1 (100%) 

Lymph node, mandibular 

-- Hemorrhage 

, . (lO) 

3 (30%) 

(10) ~ (10) 

2 (20%) 

Pigmentation 

Lymph nMe, mesenteric 

: 

: 

1 (10%) 

(lO) (10) (lO) 

Hemorrhage - 1 (1o%) 

Hyperplasia, lymphoid 1 (10%) 

Pigmentation .. - 1 (10%) 

Spleen ., " " (lO) (10) (lO) 

Hematnpoiefic cell proliferation t 00%) 3 (30%) 2 (20%) . . . . .,.  ̧... 

Pigmentation, hemosiderin 2 (20%) 4 (40%) 

Thymus (9) (1o) (10) 

Hemorrhage I (11%) • 

: 

.,, Integumentary System 

Mammary gland (9) - (8) 0o) 

Hypeyplasia, cystic 1 (11%) 1 (t3%) 
:~ ~.---__~-~-~ : 7~-~r-~ ~7-~~ z.~-~---~-............................ 

I . 

Die~ry IRestriction, NT~ TR 46~ 265 ..... = 

TABLE E3a • 

Summary of the Incidence of Nonneo#~stic Lesions in Mate lRats in the Dietary ~estriction Study 

of S~d~cy~z~su~fapyridine: Ad Libitum l~eeding and Weight-Matched Controls ,l~hrot~co~s (continued) 

:, 

Ad Libitum- Weight-Matched 

Fed Control Control 337.5 mg/kg 

1S-Month Ynter~m EvaluaKon (continued) 

Respiratory System 

Lung (10) (10) (I0) i 

Hemorrhage 1 (10%) 

Infdtration cellular, histioeyte 2 (20%) 2 (20%) 

Inflammation, subacute 5 (50%) 2 (20%) 

Alveolar epithelium, hyperplasia 3 (30%) 1 (10%) 

Nose (lO) (10) (1o) 

Exudate 2 (20%) 1 (lO%) i -

Foreign body 1 (10%) • 

; 

: 

I 

Urinary System 

Kidney 

Mineralization 

(10) (lo) 

2 (20%) 

(10) 

Nephropathy 10 (100%) 10 (100%) 10 (100%) I 7 

Renal tubule; dilatation 1 (10%) 2 (20%) • 

Transitional epithelium, hyperplasia 1 (10%) ~ ;~ 

I ~ 

Systems Examined With No Lesions Observed 

General Body System i 

Mnsculoskeletal System i Nervous System 

Special Senses System 

j: 

2- ~" ear Study 

Alimentary System 

Esophagus (50) (49) (49) 

Perforation 1 (2%) 1 (2%) 

Intestine large, colon (49) (50) (48) 

Inflammation, subacute 1 (2%) 

Parasite metazoan 7 (14%) 5 (lO%) s (1o%) 

Intestine large, rectum (50) (50) (49) 

Parasite mctazoan I0 (20%) 6 (12%) S (1o%) 

Ulcer 1 (2%) 

Intestine large, cecum (5o) (50) (50) 

Dilatation I (2%) 

Edema .. 2 (4%) 

Inflammation, subacute 1 (2%) 

Parasite metazoan 1 (2%) 

Ulcer 1 (2%) • 

Intestine small, duodenum (SO) (50) (50) 

Erosion 1 (2%) 

Ulcer 1 (2%), 

Intestine small, ileum (49) (50) ~5o) 

Inflammation, chronicactive '1 (2%) 

Ulcer (4~) 
266 Dietary Restriction, NTP TR 460 

TArtLY. E3a 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary,Restriction Study 

of S~icylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Alimentary System (continued) 

Liver 

Basnphilic focus 

Clear cell focus 

Congestion 

Degeneration, cystic 

Developmental malformation 

Eosinophilic focus 

Hematopoietic cell proliferation 

Hepatodiaphragmatic nodule 

Inflammation, subacute 

Mixed cell focus 

Bile duct, hyperplasia 

Centrilobular, atrophy 

Hepatocyte, va6uolization cytoplasmic 

Lobules, necrosis 

Mesentery 

Accessory spleen 

Fat, necrosis 

Pancreas 

Atrophy 

Acinar cell, basophilic focus 

Acinar cell, cytoplasmic alteration 

Acinar cell, hyperplasia, focal 

Salivary glands 

Atrophy 

Stomach, forestomach 

Edem~ 

Erosion . 

Infla~aaation, chronic 

Mineralization 

Perforation 

Ulcer 

Mucosa, hyperplasia 

Stomach, glandular 

Cyst 

Erosion 

Ulcer 

.. 

Cardiow~scular System 

Blood vessel 

Embolus 

Hypertrophy 

Inflammation, chronic active 

Heart 

Cardiomyopathy 

Foreign body 

Inflammation, subacute 

Mineralization 

Pericardium, fibrosis 

Ad LibitumFed Control 

(50) 

33 (66%) 

32 (64%) 

I (2%) 

4 (8%) 

7 (14%) 

5 (10%) 

5 (10%) 

45 (90%) 

3 (6%) 

2 (4%) 

(17) 

14 (82%) 

(5O) 

18 (36%) 

1 (2%) 

I (2%) 

16 (32%) 

(50) 

(50) 

2 (2%) 

1 (2%) 

(49) 

1 (2%) 

1 (2%) 

Weight-Matched 

Control 

(50) 

36 (72%) 

21 (42%) 

2 (4%) 

7 (14%) 

2 (4%) 

3 (6%)• 

2 (4%) 

21 (22%) 

40 (80%) 

4 (8%) 

3 (6%) 

(16) 

1 (6%) 

14 (88%) 

(50) 

17 (34%) 

3 (6%) 

2 (4%) 

17 (34%) 

(50) 

(50) 

1 (2%) 

2 (4%) 

(50) 

I(2%) 

(50) (50) 

l (2%) 

I (2%) 

I (2%) 

(50) (50) 

23 (46%) 30 (60%) 

1 (2%) 

337.5 mg/kg 

(50) 

20 (40%) 

16 (32%) 

1 (2%) 

9 (18%) 

2 (2%) 

2 (4%) 

2 (4%) 

2 (4%) 

21 (42%) 

2 (4%) 

1 (2%) 

(12) 

1 (8%) 

9 (75%) 

(50) 

19 (38%) 

15 (30%) 

8 (16%) 

(50) 

I (2%) 

(50) 

4 (8%) 

2 (4%) 

I (2%) 

I (2%) 

I (2%) 

7 (14%) 

9 (28%) 

(50) 

• " l (2%) 

2 (4%) 

I (2%) 

(5O) 

5 (10%) 

2 (4%) 

(50) 

24 (48%) 

.1 (2%) 

2 (2%) 

2 (4%) 
I 

. . . . . . . _. . . . . ...... ........ ...... ................. 

Dietary Restriction, NT~ TR ~0 267 

T~d~LE E3a 

S~mary of the l~ncldence o~ Nonneoplmfic Lesions in Ma~e Rats in the Dietary]~estricfion Stagy 

o~ Sa~icylaz~sa~fapyridine: Ad Libitum Feeding ~nd We~ght-I~atched Controls ~otoco~s ~continued) 

Ad Libitum~ Weight-Matched 

Fed Control Control 337.5 mg/kg 

2- ~" ear Study (continued) 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Basophilic focus 

Congestion 

Cyst 

Degeneration, fatty 

Hematopeietic cell proliferation 

Hemorrhage 

Hyperplasia, focal 

Hypertrophy, focal 

Adrenal medulla 

Hyperplasia 

Islets, pancreatic 

Hyperplasia 

Parathyroid gland 

Hyperplasia 

Pituitary gland 

Congestion 

Pars distalis, angiectasis 

Pars distalis, cyst 

Pars distalis, hyperplasia, focal 

Pars intermedia, angiectasis 

Pars intermedia, cyst 

Thyroid gland 

Ultimobranehial cyst 

C-cell, hyperplasia 

Follicle, cyst 

Follicular cell, hyperplasia 

Follicular cell, hypertrophy 

(50) 

8 (16%) 

1 (2%) 

8 (16%) 

2 (4%) 

i (2%) 

3 (6%) 

(49) 

14 (29%) 

(50) 

1 (2%) 

"(45) 

1 (2%) 

(49) 

2 (4%) 

5 (lO%) 

12 (24%) 

I (2,%), 

2 (4%) 

(50) 

1 (2%) 

11 (22%) 

(2%) 

(8%) 

(50) 

19 (38%) 

1 (2%) 

1 (2%) 

8 (16%) 

2 (4%) 

4 (8%) 

5 (10%) 

(50) 

15 (30%) 

(50) 

(46) 

(50) 

1 (2%) 

4 (8%) 

5 (10%) 

15 (30%) 

2 (4%) 

(50) 

1 (2%) 

14 (28%) 

1 (2%) 

1 (2%) 

1 (2%) 

(50) 

13 (26%) 

4 (8%) 

1 (2%) 

3 (6%) 

2 (4%) 

(50) 

14 (28%) 

(50) 

2 (4%) 

(50) 

(48) 

4 (8%) 

6 (13%) 

14 (29%) 

1 (2%) 

(50) 

3 (6%) 

14 (28%) 

1 (2%) 

~ ~£ 

i s? 

General Body System 

None 

Genital System 

Epididymis 

Atypia cellular 

Hypospermia 

Preputial gland 

Ectasia 

Hyperplasia 

Inflammation, chronic 

Inflammation, suppurative 

Prostate 

Corpora amylacea 

Edema 

Hemorrhage 

Hyperplasia, lymphoid 

Inflammation, suppurative 

Epithelium, hyperplasia 

(50) 

31 (62%) 

36 (72%) 

(50) 

14 (28%) 

2 (4%) 

9 (18%) 

17 (34%) 

(5o) 

11 (22%) 

2 (4%) 

3 (6%) 

21 (42%) 

3 (6%) , 

(50) (50) 

25 (50%) .... 32 (64%) 

40 (80%) 33 (66%) 

(50) (50) 

20 (40%) 12 (24%) 

1 (2%) 1 (2%) 

24 (48%) 16 (32%) 

19 (38%) 14 (28%) 

(50) (50) 

28 (56%) 22 (44%) 

1 (2%) 

17 (34%) 27 (54%) 

8 (16%) 8 (16%) 
268 .Dietary Restriction, NTP TR 460 

TAn,LE E3a 

Summary of the Incidence of Nonneoplastic Lesionsin Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Genital System (continued) 

Semimd vesicle 

Dilatation 

Hemorrhage 

Inflammation, suppurative 

Testes 

Granuloma sperm 

Interstitial cell; hypcrplasia 

Scminiferous tubule, atrophy 

Hematopoietic System 

Bone marrow 

Hypcrcellularity 

Lymph node 

Ifiac, pigmentation 

Mediastinal, hemorrhage 

Mediastinai, hyperplasiar lymphoid 

Mediastinal, pigmentation 

Pancreatic, hemorrhage 

Pancreatic, hypcrplasia, lymphoid . 

Renal,, pigmentation 

Lymph node, mandibular 

Congestion 

Ectasia 

Hemorrhage 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Pigmentation 

Lymph node, mesenteric 

Ectasia 

Hemorrhage 

Hyperplasia, lymphoid 

Pigmentation 

Spleen 

Fibrosis ~ 

Hematopoietic cell proliferation 

Hyperplasia, reticulum cell 

Metaplasia 

Necrosis 

Pigmentation, hemosiderin 

LYmPhoid follicle, atrophyThymus .i, 

~Hemorrhagc 

Cortex, atrophy ~. 

Ad LibitumFed Control 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

3 (6%) 

(9) 

6 (67%) 

5 (56%) 

-i 

(50) 

5 (10%) 

4 (8%) z 5 (10%) .... 

1 (2%) 

(50) 

3 (6%) 

1 (2%) 

(50) " 

8 (16%) 

14 (28%) 

14 (28%) 

(47) ; 

Weight-Matched 

Control • 

(50) 

2 (4%) 

(50) 

1 (2%) 

7 (14%) 

4 (8%) 

(50) 

3 (6%) 

(15) 

1 (7%) 

,6 (40%) 

8 (53%) 

1 (7%) 

1 (7%) 

I (7%) 

(49) 

1 (2%) 

7 (14%) 

4 (8%) 

5 0o%) ' 

I (2%) 

2 (4%) 

(50). 

2 (4%) 

6 (12%) 

1 (2%) 

2 (4%) 

(50) 

• 3 (6%) 

9 08%) 

1 .(2%) 

~ (2%) 

20 (4o~) 

,. 1 .(2%) • . 

(50) 

-~1 (2~) 

: , 

337.5 mg/kg 

(49) 

4 (8%) 

2 (4%) 

4 (8%) 

(50) 

1 (2%) 

2 (4%) 

(50) 

5 (1o%) ~ 

(1o) 

4 (40%) 

1 (10%) 

5 (50%) 

(49) 

1 (2%) : 

3 (6%) 

7 (14%) • 

(49) 

4 (8%) 

, 

(5(~ (4%), ...... ') 

23 (46%) ........ 

1 (2%) 

,::.,•• -, •, ;. . 

•3O (6O%) 

5 (10%) ..... : , 

(49) ' ,.. ~i. 

1 (2%) - ' ,,:-, 

2 (4%) :.. .,., 

-,,. ~ 

• 
i 

Dietary Restriction, N'r~ TR 460 269 

TAeL~ E3a 

Semn~ary of the ~ncidence of Nonneoplzstic Lesions in Ma~e Rats in the Dietary Restriction Study i 

o~ S~icy~zosul~apyridine: Ad Libitum Feeding and Weight-Matched Controls I~rotoco~s (continued) 

Ad Libitura- Weight-Matched 

Fed Control Control 337.5 mg/kg 

l 

I 

2-Year Study (continued) 

l(ntegumentary System 

Mammary gland 

Hyperplasia, cystic 

Hyperplasia, lobular 

Start 

Acanthosis 

Cyst epithelial inclusion 

Hemorrhage 

Hyperkeratosis 

Inflammation, chronic 

Inflammation, suppurative 

(48) 

11 (23%) 

2 (4%) 

(50) 

2 (4%) 

1 (2%) 

1 (2%) 

(50) 

16 (32%) 

I (2%) 

(50) 

2 (4%) 

4 (8%) 

2 (4%) 

I (2%) 

(49) 

10 (20%) 

6 (12%) 

(50) 

1 (2%) 

1 (2%) 

! ? 

7 

I~ 

Musculoskeletal System 

Bone 

Calvarium, osteopetrosis 

(50) (50) 

1 (2%) 

(50) 

1 (2%) 

i 

Nervous System • ~ 7 

Brain (50) (50) (50) 

Atrophy 

Compression 4 (8%) 

1 (2%) 

7 (14%) 4 (8%) 

Hydrocephalus 1 (2%) 5 (lO%) 2 (4%) 

Mineralization I (2%) 

Necrosis 1 (2%) 

Respiratory System 

Lung 

Congestion 

Edema 

Fibrosis 

Foreign body 

Hemorrhage 

Infiltration cellular, histiocyte 

Inflammation, suhacute 

Inflammation, suppurative 

(50) 

1 (2%) 

8 06%) 

3 (6%) 

(50) 

2 

2 

2 

5 

8 

3 

2 

(4%) 

(4%) 

(4%) 

(10%) 

06%) 

(6%) 

(4%) 

(50) 

4 

4 

1 

1 

12 

3 

(8%) 

(8%) 

(2%) 

(2%) 

(24%) 

(6%) 

J 

Metaplasia, osseous 

Alveolar epithelium, hyperplasia 

Fat, mediastinum, necrosis 

10 (20%) 

I (2%) 

7 (14%) 

1 

6 

(2%) 

02%) 

Nose 

Exudate 

Foreign body 

Fungus 

Mucosa, hyperplasia 

(50) 

15 (30%) 

4 (8%) 

6 (12%) 

6 (12%) 

(50) 

15 

7 

3 

8 

, 

(30%) 

(14%) 

(6%) 

(16%) 

(50) 

14 

2 

2 

6 

(28%) 

(4%) 

(4%) 

(12%) 

Mucosa, metaplasia, squamous I ..(2%) .,3 (6%) 1 (2%) 
270 Dietary Restriction, NTP TR 460 

TABLE l~3a 

Summau~ of the Incidence of Nonneoplastic Lesions in Male Rats in r the Dietary Restriction Study 

of Salicylazosuffapyridine: Ad Libitum Feeding and Weight.Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

• Fed Control Control 337.5 mg/kg 

2-Year Study (continued) 

Special Senses System 

None 

Urinary Sy.qem 

Kidney (50) (50) (50) 

Concretion 33 (66%) 

Cyst 1 (2%) I (2%) 

Hemorrhage I (2%) 

Hydronephrosis 28 (56%) 

Infarct 

Inflammation, suppurative 6 (12%) 3 (6%) 

l (2%) 

6 (12%) 

Mineralization 3.(6%) ~ 6 (12%) 13 (26%) 

Nephropathy 48 (96%) 50 (100%) 48 (96%) 

Papilla, fibrosis I (2%) 

Papilla, necrosis 4 (8%) 

Renal tubule, dilatation 1 (2%) 11 (22%) 

Renal tubule, pigmentation 2 (496) 3 (6%) 3 (6%) 

Transitional epithelium, hyperplasia 10 (20%) 5 (10%) 43 (86%) 

Transitional epithelium, hyperplasia, 

atypical 3 (6%) 

Transitional epithelium, metaplasia I (2%) 

Ureter (13) 

Concretion 3 (23 %) 

Dilatation 2 (15%) 

Inflammation, subacute 1 (8%) 

Mucosa, hylx;rplasia 

Urethra (1) 

12 

(5) 

(922%) 

Concretion ~ .1 (20%) 

Inflammation, subacute 1 (20%) 

Bulbourethral gland, ectasia I OO0%) 

Lumen, concretion 2 (40%) 

Mucosa, hyperplasia I (20%) 

Mucosa, metaplasia, squamous 2 (40%) 

Urinary bladder (50) (50) (50) 

Concretion 10 (20%) 

Congestion I (2%) 

Dilatation 1 (2%) 7 (14%): 

Hemorrhage 

Inflammation, subacute 

4 (8%), 

3 (6%) 

Mucosa, hypetplasia 41 (82%) 

. . . . . . . . . . . . . • 
Die~ry Restriction, ~ T~ ~I~ ~71

TABLE E3b 

Sun~n~ry o~ the llnc[dence o~ Nonneop~st~c Lesions in M~e Rats in the D~et~ry Restriction Study 

o~ Salicyla~osul~apyridline: :l-Year and 30-Month l~estr~cte~ Feed ][~rotoco~s a 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.S mg/kg Control 337.Smg/kg 

Disposition Summary 

Animals initially in study 61 60 49 50 

15-Month interim evalualion 10 10 

Early deaths 

Accidental deaths 2 2 4 2 

Moribund 13 6 28 14 

Natural deaths 2 3 7 10 

Survivors I Died last week of study 1 

Terminal sacrifice 34 39 10 23 

Animals examined microscopically 61 60 49 50 ]~ 

i ; 

i 

15-Month ~nter~m Evaluation 

Alimentary System 

Intestine large, colon (10) (10) 

Parasite metazoan 2 (20%) 

Intestine large, rectum (10) (10) 

Parasite metazoan • 1 (10%) 

Liver (10) (10) 

Basophilic focus 1 (10%) 

Clear cell focus 1 (10%) 

Congestion 1 (10%) 

Eosinophilic focus 1 (10%) 3 (30%) 

Hepatodiaphragmatic nodule 2 (20%) 

MiXed cell focus 3 (30%) 3 (30%) 

Bile duct, hyperplasia 3 (30%) 

Hepatocyte, vacuolization cytoplasmic 5 (50%) 

Kupffer cell, pigmentation 1 (10%) 

Lobules, necrosis 1 (10%) 

Mesentery (1) (1) 

Fibrosis 1 (100%) 

Fat, necrosis 1 (100%) . 

Pancreas (10) (10) 

Atrophy 5 (50%) 3 (30%) 

Pigmentation 1 (10%) 

Acinar cell, cytoplasmic alteration 1 (10%) 

Aeinar cell, hyperplasia, focal 2 (20%) I (10%) 

Stomach, forestomach (10) (10) 

Erosion I (10%) 

Mineralization 2 (20%) 

Ulcer 2 (20%) 

Mucosa, hyperplasia 1 (10%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
272 Dietary Restriction, NTP TR 460 

TABLE E3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30.Month Restricted Feed 

Vehicle 

Control • 

15-Month Interim Evaluation (continued) 

Cardiovascular System 

Heart (10) 

Cardiomyopathy 4 (40%) 

Inflammation, subacute 1 (10%) 

Endocrine System 

Adrenal cortex (10) 

Accessory adrenal cortical nodule 2 (20%) 

Hype~plasia, focal 

Islets, pancreatic (10) 

Hyperplasia 

Pituitary gland (I0) 

Pars distalis, hyperplasia, focal 2 (20%) 

Thyroid gland (10) 

C-cell, hyperplasia 

Genital System . .. 

Epididymis . 

(10) 

Atypia cellular 4 (40%) 

Hypospermia 1 (10%) 

Preputial gland (10) 

Inflamrnation, chronic 7 (70%) 

Prostate (10) 

Corpora amylacea 2 (20%) 

Inflammation, suppurative • 4 (40%) 

Testes " (10) 

Interstitial cell, hyperplasia 2 (20%) 

Seminiferous tubule, atrophy 2 (20%) 

Hematopoietic System 

Lymph node (2) 

Mediastinal, hemorrhage 2 (I00%) 

Lymph node, mandibular (lo) 

Hemorrhage 

Lymph node, mesenteric 0o) 

Hemorrhage 1 0o%) 

Spleen (lo) 

Pigmentation, hemosiderin 1 0o%) 

Integumentary System 

Mammary gland (10) 

Hyperplasia, cystic 1 (10%) 

Vehicle 

337.5 mg/kg Control 337.5 mg/kg 

Oo) 

4 (40%) 

(10) 

1 (10%) 

1 (10%) 

(10) 

1 (10%) 

(I0) 

1 (10%) 

(lO) 

1 (lo%) 

(10) 

2 (2O%) 

1 (10%) 

(I0) 

4 (40%) 

(10) . 

2 (20%) 

4 (40%) 

(lo) 

6 (6O%) 

(lO) 

1 (10%) 

(lO) 

(lo) 

5 (50%) 

(9) 
2 

:::: .......................... : : ........................ =-:--_~-~- ~-----.~.~ -__ _.~ ~ ~_--~.:-----___ 

Dietary Restriction, NTP TR 460 273 

TABLE E3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

o~ SaHicy~azosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

i : ~? 

1 

i 

I 

~2--

~ 2:

: 

; 

i 

~ _ 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5mg/kg 

15-Month lrnterim Evaluation (continued) 

I Respiratory System 

Lung (10) (10) t = 

Hemorrhage 

Infdtration cellular, histiocyte 1 (10%) 

1 

1 

(10%) 

(10%) 

! 

[! 

i 

: 

Inflammation, subacute 1 (10%) 

Alveolar epithelium, hyperplasia 1 (10%) 1 (lO%) ! 

Nose (10) (10) 

Exudate 1 (10%) 

Fungus 1 (10%) 

Mucosa, metaplasia, squamous 1 (10%) i ~ 

Urinary System [ 

Kidney 

Concretion 

(10) (10) 

3 (30%) i

i' ~ 

Nephropathy 

Transitional epithelium, hyperplasia 

Urethra 

Bulbourethral gland, ectasia 

10 (100%) 

(1) 

1 (100%) 

10 (lOO%) 

1 (10%) '!! 

! 

[~ 

! 

• 

l 

i 

Systems Examined With No Lesions Observed 

General Body System 

Musculoskeletal System 

i 

i 

i 

i

1 

Nervous System i 

i: 

Special Senses System i 

2-Year and 30-Month Protocols 

Alimentary System 

~ I Esophagus 

Foreign body 

(50) (50) 

1 (2%) 

(49) (48) 

Hemorrhage 1 (2%) 1 (2%) 

Inflammation, granulomatous 1 (2%) 

Inflammation, suppurative 1 (2%) 1 (2%) 

Intestine large, colon (51) (49) (49) (47) 

Dilatation 1 (2%) 1 (2%) 

Parasite metazoan 8 (16%) 6 (12%) 2 (4%) 

Intestine large, rectum (51) (50) (49) (49) 

Edema 1 (2%) 

Parasite metazoan 9 (18%) 6 (12%) 7 (14%) 2 (4%) " 

Ulcer 1~ (2%) 
274 Dietary Restriction, NTP TR 460 

TABLE E3b t 

Summary r of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Salicy]azosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

2-Year and 30-Month Protocols (continued) 

Alhnentary System (continued) 

Intestine large, cecum (51) (50) (49) (49) 

Dilatation 

Edema 

1 (2%) 

I (2%). 

i (2%) 

1 (2%) 

Hemorrhage I (2%) 

Parasite metazoan 2 (4%) 1 (2%) 

Ulcer 2 (4%) 

Intestine small, duodenum (50) (50) (49) (49) 

Erosion 1 (2%) 

Liver (51) (50) (49) (50) 

Angiectasis 1 (2%) 2 (4%) 

Basophilic focus 24 (47%) 5 (10%) 16 (33%) 11 (22%) 

Clear cell focus 11 (22%) 5 (10%) I (2%) 

Congestion I (2%) 1 (2%) 

Concretion (2%) 

Cyst i (2%) 

Degeneration, cystic 2 (4%) • . 

• Developmental malformation 1 (2%) 

Eosinophilic focus 9(18%) 15 (30%) (16%) 6 (12%) ' 

Hematopoietie cell proliferation (2%) 1 (2%) 

Hepatodiaphragmatic nodule I (2%) 6 (12%) (8%) I (2%) 

Inflammation, chronic 1 (2%) (2%) I (2%). 

Inflammation, subacute 1 (2%) 

Mixed cell focus 17 (33%) I (2%) 2 (4%) 3 (6%) 

Bile duct, hyperplasia - 

Centrilobular, atrophy 

36 (71%) 

3 (6%) 

27 (54%) 35 

12 

01%) 

(24%) 

35 (70%) 

4 (8%) 

Centrilobular, necrosis I (2%) , 

Hepatocyte, vaeuolization cytoplasmic 2 (4%) (12%) 

Hepatocyte, vaeuolization nuclear 1 (2%) 

Kupffer cell, hyperplasia 1 (2%) 

Kupffer cell, pigmentation 2 (4%) 7 (14%) ... 

Lobules, necrosis I (2%) 3 (6%) I (2%) 

Mesentery (20) (7) (! 4 ) (10) 

Accessory spleen 1 (14%) I (7%) I (10%) 

Fat, necrosis 18 (9O%) 5 (71%) 9 (64%) 4 (4o%) 

Pancreas (51) (50) (49) (48) 

Atrophy 19 (37%) 16 (32%) 21 (43%) 17 (35%) 

Metaplasia, hepatocyte 1 (2%) • . 

Necrosis 

Acinar cell, cytoplasmic alteration 

Acinar cell, hyperplasia 

Aeinar cell, hyperplasia, focal 

I (2%) 

3 (6%) 

8 (16%) 

1 (2%) 

3 (6%) 

(2%) 

(2%) 

(12%) 

1 

1 

6 

2 (4%) 

'2 (4%) 

Salivary glands (51) (50) (49) (49) 

Atrophy 3 (6%) 

Inflammation,, chronic I (2%) 
I 

i 

- _ _ ..... ~ _ _ _ _ 

!~ : ~ 7 ~ ~ ........ ~--__-~ .. ............................. 

Dietary ~es~'lcfion, ~ T~ 46~ 

T~L~ ~3b 

S~ary ot ~ the ~ucidence o~ Nonneop~astic Lesions in ~a~e Rats in the Dietary Restriction Study. 

o~ Sa~Jcy~azosu~apy~d~e: 2-Year and 3~I~Iont~ Restricted Feed ~'otocols (continued). 

2-Year ]Restricted Feed 30-Month ]Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

2-3"ear and 30-Mong1~ Protocols (continued) 

Alimentary System (continued) 

Stomach, forestomach (51) (50) (49) (50) 

Developmental malformation 1 (2 %) 

Erosion 1 (2%) 

Inflammation, chronic 1 (2%) 

Inflammation, suppurative 1 (2%) 1 (2%) 1 (2%) 

Mineralization 2 (4%) 1 (2%) 1 (2%) 2 (4%) 

Ulcer 1 (2%) 2 (4%) 

Mucosa, hyperplasia 4 (8%) 3 (6%) 3 (6%) 

Stomach, glandular (51) (50) (49) (50) 

Edema 1 (2%) 1 (2%) 1 (2%) 

Erosion 1 (2%) 2 (4%) 

Fibrosis 1 (2%) 

Inflammation, suppurative I (2%) 

Mineralization 1 (2%) 

Ulcer (2%) 1 (2%) 

Cardiovascular System 

Blood vessel (51) (50) (48) (50) 

Hypertrophy 1 (2%) 1 (2%) 1 (2%) 

Inflammation, chronic active 1 (2%) 1 (2%) 1 (2%) 

Aorta, mineralization 1 (2%) 

Heart (51) (50) (49) (49) 

Cardiomyopathy 33 (65%) 23 (46%) 36 (73%) 32 (65%) 

Inflammation, subaeute 1 (2%) 1 (2%) 

Mineralization 1 (2%) 

Thrombosis 3 (6%) 

Endocardium, hyperplasia 1 (2%) 

Endocrine System 

Adrenal cortex (51) (50) (49) (50) 

Accessory adrenal cortical nodule 15 (29%) 12 (24%) 11 (22%) 15 (30%) 

Angiectasis 1 (2%) 3 (6%) 3 (6%) 

Atrophy 1 (2%) 

Cyst 1 (2%) (4%) 

Degeneration, fatty 6 (12%) 3 (6%) 2 (4%) (14%) 

Hemorrhage 1 (2%) 1 (2%) 

Hyperplasia, focal 1 (2%) 1 (2%) 4 (8%) 

Hypertrophy, focal 1 (2%) 3 (6%) 2 (4%) (4%) 

Mineralization, focal 1 (2%) 

Necrosis 1 (2%) 

Adrenal medulla (51) (50) (48) (50) 

Hyperplasia 6 (12%) 3 (6%) 11 (23%) 3 (6%) 

Islets, pancreatic (51) (50) (49) . (48) 

Hyperplasia 2 (4%) 

~ 

i ~ 

I : 

i 

i ~ 

275 i ' 

I 

; 

• 

! 

! ~ 

[ i 

I 

i 

1 

I ~ !: 

i z 

i 

~ . 

i 

:; 

!~ 

!: 

{: 

i = 

, 

• : 

r 

i 

i 
276 Dietary Restriction, NTP TR 460 

TABLE E3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the Dietary Restriction Study 

of Salicy]azosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

2-Year and 30-Monih Protocols (continued) 

Endocrine System (cominued) 

Parathyroid gland 

Hyperplasia 

(48) (47) 

I (2%) 

(48) 

Pituitary gland 

Pars distalis, angiectasis 

(51) 

4 (8%) 

(49) 

4 (8%j 

(49) " 

1 (2%) 

(49) 

3 (6%) 

Pars distalis, cyst 7 (14%) 5 (1o%) 8 (16%) 3 (6%) 

Pars distalis, hyperplasi a 

Pars distalis, hyperplasia, focal 9 (18%) 

1 (2%) 

8 (16%) 8 (16%) 8 (16%) 

Pars-intermedia, angiectasis 1 (2%) 1 (2%) 1: (2%) 1 (2%) 

Pars intermedia, cyst 1 (2%) 4 (8%) 1 (2%) 

Thyroid gland (50) (50) (49) (49) 

Ultimobranchial cyst I (2%) 

C-cell, hyperplasia 10 (20%) 5 (10%) (6%) 9 (18%) 

Follicle, cyst 2 (4%) 2 (4%) 4 (8%) 

Follicle, mineralization 7 (14%) 38 (76%) 

Follicular cell, hyperplasia 13 (6%) 1 (2%) 

General Body System 

None 

Genital System 

Epididymis 

Atypia cellular 

Hypospermia 

Preputial gland 

(51) 

38 (75%) 

-35 (69%) 

(51) 

(50) 

33 (66%) 

34 (68%) 

(50) 

(49) 

24 (49%) 

38 (78%) 

(49) 

(49) 

25 

37 

(49) 

(51%) 

(76%) 

Ectasia 10 (20%) 8 (16%) 10 (20%) I0 (20%) 

Hyperplasia 2 (4%) 2 (4%) 

Inflammation, chronic 30 (59%) 20 (40%) 18 (37%) 10 (20%) 

Inflammation., suppurative 

Prostate 

Corpora amylacea 

Edema 

8 (16%) 

(51) 

28 (55%) 

11 (22%) 

(50) 

25 (50%) 

l (2%) 

8 (16%) 

(49) 

23 (47%) 

8 (16%) 

(49) 

11 (22%) 

1, (2%) 

Fibrosis I (2%). 

Hemorrhage 

Inflammation, suppufative 

Epithelium, hyperplasia 

Seminal vesicle 

Inflammation, suppurative 

Testes 

Interstitial cell, hyperplasia 

Seminiferous ~ubule, atrophy 

27 (53%) 

15 (29%) 

(51) 

(51) 

7 (14%) 

1 (2%) 

1 (2%) 

20 (40%) 

10 (20%) 

(50) 

1 (2%) 

(50) 

5 (10%) 

2 (4%) 

19 (39%) 

13 (27%) 

(49) 

(49) 

3 (6%) 

1 (2%) 

-- 2 (4%) 

26 (53%) 

7 (14%) 

(48) 

3 (6%) 

(49) 

3 (6%) 

5 (1o%) 
~ : ~ 

:~: ~ 

~ ~-~ -~--~-~ , ,r.~..~-~,.: ...... ,~.~ - - .................. - - - , ~ ............ ~ ................. 

J 

i ~ -

Dietary ]Restriction, NTP TR 460 277 

TABLE E3b 

Surnmm'y of the I[ncidence of Normeoplmstie Lesions in Ma~e Rats in the Dietary l~estr~ction Study 

of Sa]icy~zosugapyridine: 2-'Year and 30-Month Restricted Feed Protocols (continued) , • _ 

2-Year Restricted Feed 30-Month Restricted Feed 

Vehicle Vehicle 

Control 337.5 mg/kg Control 337.5 mg/kg 

2-Year and 30-Month Protocols (continued) 

Hematopoietic System 

Bone marrow (51) (50) (49) (50) 

{ Hypercellularity 

Myelofibrosis 

2 (4%) 2 (4%) 

4 (8%) 

4 (8%) 

1 (2%) 

Lymph node (11) (3) (20) (10) i : 

Axillary, hemorrhage 1 (10%) 

Iliac, hemorrhage I (10%) 

Iliac, hyperplasia, lymphoid 1 (10%) 

lliac, pigmentation 1 (10%) 

Inguinal, hemorrhage 1 (9%) 1 (33%) 1 (5%) 

Mediastinal, embolus 

Mediastinal, hemorrhage 2 (18%) 2 (67%) 5 (25%) 

1 

I 

(10%) 

(10%) 

Mediastinal, hyperplasia, lymphoid 1 (9%) 1 (5%) 

Mediastinal, infiltration cellular, mast cell 1 (9%) 

Mediastinal, pigmefitation 4 (36%) 1 (33%) 7 (35%) 3 (30%) 

Pancreatic, ectasia 1 (5%) 

Pancreatic, hemorrhage 2 (2O%) 

Pancreatic, pigmentation 

Lymph node, mandibular 

2 (18%) 

(51) (50) (48) 

1 

(49) 

(10%) i

i 

Congestion 

Ectasia 

Hemorrhage 

3 (6%) 

3 (6%) 

2 (4%) 

9 (18%) 

1 (2%) 

2 (4%) 

1 

4 

(2%) 

(8%) 

Hyperplasia, lymphoid 1 (2%) 2 (4%) 7 (15%) 4 (8%) 

Pigmentation 

Lymph node, mesenteric 

7 (14%) 

(51) 

6 (12%) 

(50) 

4 (8%) 

(49) 

2 

(48) 

(4%) 

Ectasia 1 (2%) 

Hemorrhage 

Hyperplasia, lymphoid 

Pigmentation 

4 (8%) 

2 (4%) 

4 (8%) 

3 (6%) 5 (10%) 

3 (6%) 

(15%) 

(2%) 

Spleen (51) (50) (49) (49) 

Fibrosis 3 (6%) I (2%) ;3 (27%) 2 (4%) 

Hematopoietic cell proliferation 6 (12%) 6 (12%) 9 (18%) 13 (27%) 

Hemorrhage 3 (6%) 2 (4%) 

Pigmentation, hemosiderin 12 (24%) 35 (70%) 15 (31%) 33 (67%) 

Lymphoid follicle, atrophy 1 (2%) 

Thymus (47) (48) (44) (40) 

Cyst 1 (2%) 

Hemorrhage 1 (2%) 2 (4%) 

Cortex, atrophy 1 (2%) 
278 .. Dietary Restriction, NTP TR 460 

TAmJ~ E3b 

Sununary of the Incidence of Nonneoplastic Lesions in Male Rats in.the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

2-Year and 30-Month Protocols, 

Integumentary System 

Manm3ary gland 

Hyperplasia, cystic 

Hyperplasia, Iobular. 

Skin 

Acanthosis 

Angiectasis 

Cyst epithelial inclusion 

Hemorrhage 

Hyperkeratosis 

Inflammation, chronic 

Inflm~mation, suppurative 

Subcutaneous tissue, edema 

MuscuJ~skeletai System 

Bone 

Calvarium, osteopetrosis 

Femur, osteopetrosis 

Skeletal muscle 

Hemorrhage 

Infla~unation, suppurative 

Necrosis 

Nervous System 

Brain 

Compress/on 

Giiosis 

HemmThage 

Hydrocephalus 

Necrosis 

Respiratory System 

Lung 

Congestion 

Edema 

Embolus 

Fibrosis 

Foreign body 

Hemorrhage 

,.hffiltration cellular, histio~yte 

Inflammation, suhacute 

Alveolar epithelium, hyperplasia 

30-Month Restricted Feed 

Vehicle 

Control 337.s mg/kg 

(49) (47) 

19 (39%) 9 (19%) 

5 (10%) 4 (9%) 

(49) (50) 

2 (4%) 5 (10%) 

1 (2%) 

I (2%) 

1 (2%) 

4 (8%) 8 (16%) 

I (2%) 

2 (4%) 

(49) (50) 

1 (2%) 

2 (4%) I (2%) 

.. 

(49) (50) 

7 (14%) 5 (10%) 

,1 (2%) 

1 (2%) 2 (4%) 

I (2%) 3 (6%) 

1 (2%) 1 (2%) 

(49) (49) 

4 (8%) 2 (4%) 

1 (2%) 2 (4%) 

I (2%) 

1 (2%) 

3 (6%) I (2%) 

3 (6%) I (2%) 

11 (22%) 18 (37%) 

3 (6%) 3 (6%) 

4 (8%) 3 (6%) 

Vehicle 

Control 

continued) 

(50) 

12 (24%) 

(51) 

1 (2%) 

I (2%) 

1 (2%) 

1 (2%) 

(51) 

(2) 

1 (50%) 

1 (50%) 

I (50%) 

(51) 

5 (1o%) 

i (2%) 

1 (2%) 

2 (4%) 

(51) 

3 (6%) 

2 (4%) 

4 (8%) 

2 (4%) 

I0 (20%) 

1 (2%) 

5 (10%) 

337.5 mg/kg 

(49) 

7 (14%) 

(50) 

1 (2%) 

1 (2%) 

I (2%) 

(50) 

I (2%) 

1 (2%) 

(50) 

2 (4%) 

1 (2%) 

I (2%) 

(50) 

4 (8%) 

3 (6%) 

4 (8%) 

1 (2%) 

9 (18%) 

1 (2%) 

5 (10%) 
i 

;: .~:~::=,,:~ ~-~,_~_~==~ ~ ~ ~ _ ~__~ .......... ~ ~ . ~ ~ . ....... . ............... _~:_ _ ~

Die~ry II~esCHc~on, ~ TR ~ .279 

2 

TA~L~ E3b 

Smma~ o~ the ~Mc~flence o~ Nonneoplast[c Lesions in MM~e ~a~s in the Die~m'y ~es~Hcfion Study 

og ~cy~agosu~apyridlne: 2-Year and 30-Momh ~estHc~ed Feed ]Protocols (continued) 

2-Year Restricted Feed 

Vehicle 

. 30-Month Restricted Feed 

Vehicle 

] 7:~ 

! 

Control 337.5 mg/kg Control 337.5 mg/kg 

I 

I 

[ 

i!~ 

I 

! 

! 

2-Year and 30-Month Protocols (continued) 

~esph'atory System (continued) 

Nose (51) 

Exudate 14 (27%) 

Foreign body 6 (12%) 

Fungus 3 (6%) 

Mucosa, hyperplasia 3 (6%) 

Mucosa, metaplasia, squamous 2 (4%) 

Trachea 

Erosion 

(50) 

10 (20%) 

2 (4%) 

3 (6%) 

4 (8%) 

1 (2%) 

(49) 

8 (16%) 

2 (4%) 

2 (4%) 

1 (2%) 

(49) 

1 (2%) 

(50) 

17 (34%) 

3 (6%) 

2 (4%) 

5 (10%) 

1 (2%) 

(49) 

i 

i 

Speci~ Senses System 

Ear 

Hyperplasia, fo~a| 

Eye 

Cataract 

Hemorrhage 

Inflammation. suppurative 

Phthisis bulbi 

Retina, atrophy 

Zymbal's gland 

Cyst 

(1) 

1 (100%) 

1 (100%) 

(2) 

1 (5O%) 

1 (50%) 

1 (50%) 

(1) 

(4) 

3 (75%) 

1 (25%) 

I (25%) 

2 (50%) 

(2) 

1 (50%) 

(1) 

1 (100%) 

(2) 

1 (50%) 

I (50%) 

• , 

t 

Orblm',y System 

Kidney 

Concretion 

Cyst 

Fibrosis 

Hemorrhage 

Hydronephrosis 

Infarct 

Inflammation, suppurative 

Mineralization 

Nephropathy 

Papilla, necrosis 

Renal tubule, cytoplasmic alteration 

Renal tubule, dilatation 

Renal tubule, necrosis 

Renal tubule, pigmentation 

Transitional epithelium, hypcrplasia 

Ureter 

Mucosa, hypcrplasia 

Urethra 

Metaplasia, squamous 

Urinary bladder 

Concretion 

Dilatation 

Hemorrhage 

Inflammation, suppurative 

Mucosa, hype~plasia 

(51 ) 

I (2%) 

2 (4%) 

44 (86%) 

I (2%) 

2 (4%) 

3 (6%) 

(5 I) 

(50) 

22 (44%) 

I (2%) 

1 (2%) 

1 (2%) 

11 (22%) 

46 (92%) 

1 (2%) 

I (2 %) 

18 (36%) 

(50) 

1 (2%) 

7 (14%) 

(49) 

1 

1 

4 

39 

1 

1 

2 

2 

1 

(2%) 

(2%) 

(8%) 

(80%) 

(2%) 

(2%) 

(4%) 

(4%) 

(2%) 

(2) 

1 

(49) 

(50%) 

I (2%) 

(50) 

35 (70%) 

I (2%) 

2 (4%) 

3 (6%) 

7 (14%) 

48 (96%) 

I (2%) 

1 (2%) 

3 (6%) 

6 (12%) 

37 04%) 

(1) 

1 (100%) 

(2) 

2 (100%) 

(49) 

1 (2%) 

2 (4%) 

2 (4%) 

2 (4%) 

8 (16%) 
Dietary Restriction, NTP TI~ 
: ~ ~ ~~ ~ _:-------- ~ _ ~ _ . ~ ~ ~ _ _ ~ ......... < ,~ , ~ , 

i 

i 

APPEH]~]Ii~ IF I 

SLZ4BJIA]~Y ©IF LES]~©HS ]~H I~IALIZ I~I]~CIZ 

]~N T]~Z ]~]~IZTA~Y ]~ESTI~]~CT]~©H 5T~/I~Y 

OF SAL]ICYLAZ©$LYL]FAP~fIZ~]~]IHE 

TABLE F~a Summary of the Incidence of Neoplasms in Male Mice 

in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls ~rotoco~s ............ ~...... Z82 

TABLE F~.b Summary of the Incidence of Neoplasms in Male Mice 

in the Dietary Restriction Study of Salicylazosulfapyridine: 

2-'Year and 3-Year Restricted Feed Protocols .............................. ~6 

TABLE F~a Statistical Analysis of Primary Neoplasms in Male Mice i < 

in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocois ................... 29~ 

TABLE ~b Statistical Analysis of Primary Neoplasms in Male Mice 

in the Dietary Restriction Study of Salicylazosulfapyridine: i ~ 

2-'Year and 3-Year Restricted Feed Protocols .............................. 295 

TABLE ]~a Summary of the Incidence of Nonneoplastlc Lesions in Male Mice 

in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocois ................... 29~ 

TABLE Y3b Summary of the Incidence of Nonneoplastic Lesions in Male Mice i 

in the Dietary Restriction Study of Salicylazosulfapyridine: 

2-'Year and 3-Year Restricted Feed Protocols .............................. 3~ 

i 

i 

i 
282 Dietary Restriction, NTP TR 460 

TAnL~ Fla 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad L/b/turn Feeding and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Control 2,700 mg/kg 

Dispo~dtion Summary 

Animals initially in study 60 60 60 

15-Month interim evaluation 10 10 10 

Early deaths 

Accidental deaths 2 1 

" Moribund 5 3 4 ~ 

Natural deaths 3 1 

Survivors 

Terminal sacrifice 40 45 

Animals ¢xamined microscopically 60 60 

15-Month Interim Evaluation' 

Alimentary System 

Liver (I0) (lo) (10) 

Hepatocellular adenoma 2 (20%) i (1o%) 2 (20%) 

Stomach, forestomach : (10) (1o) (10) 

Squamous cell papilloma 1 (10%) ,. 

Respiratory System. , 

Lung (10) (io) • • . Oo) 

Alveolar/bronchiolar adenoma I ( 10 %) 

Systems Examined-With No Neoplasms Observed 

Cardiovascular System 

Endocrine System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Mnsculoskeletal System 

Nervous System 

Special Senses System 

Urina~ System 
i 

: : - - - ? ~ ? ~ - ~ - - - ~ - ~ ~ . _ ~ _ _ ~ 

, ~__ --~z7:~-~'7~'?'~'~~ ......... ~ ~: ~ ~ :~-~'~ ~ ~ ~ : 7 ~ : ~ _ ~ ; ...... ~ ....... :~- ............--~,--~-.- .... 

Dietary Restriction, NTP TR 4~ i 

I 

t 

TABLE F1a i 

Summary o~ the ]!.nc~ence o~ l~eop~asn~s ~n Male l~'II.~ce ~a the Dietary ~estr~cct~on Stu~y 

o~ S~cy~z~sa~l~apyridline: Ad L/b/turn Feeding and Weight-l~'I~atche~ Controls ]?rot~eo~ (continued) i ~ -

i 

Ad Libitum- Weight-Matched 

~'ed Control Control 2,700 mg/kg * 

2-Year Study 

Alimentary System 

Intestine large, cecum (47) (50) (50) 

i Adenocarcinoma 1 (2%) 

Intestine small, jejunum (47) (50) (50) 

Intestine small, ileum (47) (50) 

Liver (50) (50) (50) 

1 [ Hemangioma 1 (2%) i 

i Hemangiosarcoma 2 (4%) 

I Hepatoeellular carcinoma 12 (24%) 6 (12%) 8 (16%) 

i 

[ Hepatucellular carcinoma, multiple 1 (2%) 

Hepatoeellular adenoma 12 (24%) 8 (16%) 15 (30%) 

Hepatocellular adenoma, multiple 1 (2%) 27 (54%) 

Mesentery (4) (2) (1) 

Pancreas (50) (50) (50) I 

Carcinoma, metastatic, lung 1 (2%) 

Acinus, adenoma 1 (2%) 

Stomach, forestomach (50) (50) (50) i Squamous cell papilloma 3 (6%) 5 (10%) i Stomach, glandular (50) (50) (50) j ~

Adenoma 1 (2%) 

Cardiovascular System 

None 

Endocrine System 

Adrenal cortex (50) (50) (50) 

Carcinoma, metastatic, lung 1 (2%) 

Capsule, adenoma 1 (2%) 

Adrenal medulla (50) (5o) (50) 

Carcinoma, metastatic, lung 1 (2%) 

Islets, pancreatic (50) (50) (50) 

Adenoma 1 (2%) 1 (2%) 

Pituitary gland (44) (46) (47) 

Pars distalis, adenoma 1 (2%) 

Pars intermedia, carcinoma 1 (2%) 

Thyroid gland (50) (50) (50) 

Follicular cell, adenoma 1 (2%) 2 (4%) l (2~) 

Follicular cell, carcinoma I (2%) 

General Body System 

None 

Genital System 

Preputial gland 

Adenoma " 

Prostate 

Seminal vesicle 

(49) 

(5o) 

(50) 

(50) 

(50) 

(50) 

(50) 

1 (2%) 

(50) 

(50) 
284 Dietary Restriction, NTP TR 460 

TABLE Fla 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyrldine: Ad Libitum Feeding and Weight-Matched Controls Protocol (continued) . ., 

2-Year Study (continued) 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node mesenteric ~-~ o ' Hgm~mg,osarcoma 

Spleen 

Hemangiosarcoma 

Thymus 

Carcinoma, metastatic, lung 

Integmnentary System 

Skin° 

Subcutaneous tissue, fibrosarcoma 

Subcutaneous tissue, hemangiosarcoma 

Subcutaneous tissue, plasma cell tumor benign 

Musculoskeletal System 

Skeletal muscle 

Hemangiosarcoma / 

Nervom; System 

Brain 

Respiratory System 

Lufig 

Alve01ar/bronchiolar adenoma 

Alveolar/bronchiolar adenoma, multiple 

Alveolar/bronchiolar carcinoma 

Alveolar/bronchiolar carcinoma, multiple 

Carcinoma, metastatic, tissue NOS 

Hepatocellular carcinoma, metastatic, liver 

Special Senses System 

Harderian gland 

Adenoma 

Carcinoma 

,. 

Urinary System 

Kidney 

Urinary bladder 

Hemangiioma 

Ad LibitumFed Control 

(50) 

(3) 

(50) 

(50) 

(50) 

1 (2%) 

(45) 

I (2%) 

(50) 

I (2%) 

(50) 

(50) 

9 (18%) 

2 (4%) 

3 (6%) 

4 (8%) 

(3) 

2 (67%) 

I (33%) 

(50) 

(50) 

Weight-Matched 

Control 

(50) 

(1) 

(49) 

(48) 

(50) 

1 (2%) 

(46) 

(50) 

1 (2%) 

(i) 

1 (ioo%) 

(50) 

(50) 

6 (12%) 

4 (8%) 

1 (2%) 

(3) 

2 (67%) 

(50) 

(50) 

2,700 mg/kg 

(50) " 

. (4) 

(49) 

(50) 

1 (2%) 

(50) 

.1 (2%) 

(46) 

(50) 

1 (2%) 

(50) 

(50) 

4 (8%) 

2 (4%) 

4 (8%) 

{ (2%) 

1 (2%) 

(2) 

2 (1oo%) 

(5o) 

(5o) 

I (2%) 
I 

7~y~ ~f~ ~ - ~ ~ _ . , ~

....- _ _ ~ _ _ ~ _ _ . .......... ~_. ~ _ , . _ ,, . ~ - -~ ~ ~ : ..... - - , ~ - ~:~=.~7 ~--~--=~,~.~-~!~,,~.~--~.,~-~=.~.~ ~ ~-= : ~ ~

i 

[ 

i 

Dietary ]Restriction, NT~ TR ~60 285 

i 

TA~L~ ~la 

Su~an~ary ot the ]Incidence o~ Neoplmn~s h~ Male iV~[ce in the Dietary Restriction Study 

ot Sal[cylazosul~apyridine: Ad Libitum Feeding and We[ght-IV~atchedl Controls ]?rotoeo~ (continued) 

2-Tear Study(continual) 

Systemic ~.sions 

Multiple organs b 

Lymphoma malignant 

Lymphoma malignant lymphocytic 

Neoplasm Summary 

Total animals with primary neoplasms c 

15-Month interim evaluation 

2-Year study 

Total primary neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total benign neoplasms 

~ 15-Month interim evaluation 

2-Year study 

Total animals with malignant neoplasms 

2-Year study 

Total malignant neoplasms 

2-Year study 

Total animals with metastatic neoplasms 

2-Year study 

Total metastatic neoplasms 

2-Year study 

Ad LibitumFed Control 

(50) 

5 (I0%) 

4 

40 

4 

62 

4 

26 

4 

35 

25 

27 

5 

8 

Weight-Matched 

Control 

(50) 

2 (4%) 

1 

30 

1 

38 

1 

19 

1 

21 

16 

17 

1 

1 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

¢ Primary neoplasms: all neoplasms except metastatic neoplasms 

2,7oo mg/~ 

• • 

i ~ 

(50) 

I (2%~ 

1 (2%) 

i = ̧= 

2 

45 

i

[ 

i 

i 

2 

75 

2 

43 

2 

56 

i 

14 

19 

1 

1 
286 Dietary Restriction, NTP TR 460 

TABLE Flb 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study of Salicylazosulfapyr~dine: 

2-Year aad 3-Year Restricted Feed Protocols a 

2-Year Restricted Feed 

Vehicle 

Control 2,700 mg/kg 

Disposition summary 

Animals initially in study 62 60 

15-Month interim evaluation 8 9 

Accidental deaths b 2 1 

Early deaths 

Moribund 6 

Natural deaths 4 

Survivors 

Terminal sacrifice 42 

Animals examined microscopically 62 

15-Month .Interim Evaluation 

Alimentary System 

Liver (10) (10) 

Hernangiosarcoma 1 (10%) 

Hepatocellular adenoma 1 (10%) 

Systems Examined With No Neoplasms Observed 

Cardiovascular System 

Endocrine System 

~ General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

2-Year and 3-Year Protocols 

Alimentary System 

Esophagus (52) (50) 

Gallbladder (St) (50) 

Leiomyosarc~nla, metastatic, 

stomach, glandular 

Intestine large, colon (51) (50) 

Intestine large, cecum (51) (50) 

Lymphoid tissue, leiomyoma 

Intestine small, duodenum (51) (50) 

Carcinoma, metastatic, harderian gland 1 (2%) 

• . 

3-Year Restricted Feed 

Vehicle 

Control 2,700 mg/kg 

48 50 

21 9 

7 7 

20 34 

48 50 

. 

(48) (50) 

(4s) (49) 

t (2%) 

(47) (48) 

(47) (47) 

I (2%) 

(47) (46) . 
_ _ 

= = - , .~ _. ~ - - _ _ _ ~ . ~ ............ ._~ ....... : ~ ..... . : ..... ~ ~,,~: .... ._._ ~ .... .. ~__ 

I 

Die~ry ll~estr~c~on, ~ T~ ~ ~7 I 

T~LE F~.b 

Summ~y of the ]Incidence of Neop~mms in I~,tIa~e.I~Iice in the Die~tm-y ~estricct~on St~y o~ Sa~cy~x~sug~ap]nfi~ne: 

2-Year ~nd 3-Year ~est~ccte~ Feed ]Protocols (continued) 

I 2-Year Restricted Feed 3-Year ~estr~ed Feed 

Vehicle Vehicle 

Control 2,700 mg/kg Comrol 2,7~ mg/kg 

I 

2-Year and 3-Year Protocols (continued) 

Alimentary System (continued) 

Intestine small, jejunum (51) (50) (47) (46) 

I Adenocarcinoma I (2%) 1 (2%) 

Carcinoma, metastatic, harderian gland 1 (2%) 

! Intestine small, ileum 

Liver 

(51) 

(52) 

(50) 

(50) 

(47) 

(48) 

(47) 

(50) 

Carcinoid tumor malignant, metastatic, 

stomach, glandular 1 (2%) 

Hemangiosarcoma 2 (4%) 

Hepatocellular carcinoma 6 (12 % ) 1 (2%) 13 (27%) 6 (12%) 

Hepatoeellular carcinoma, multiple 1 (2%) 3 (6%) 

Hepatoeellular adenoma 

Hepatoeellular adenoma, multiple 

Histiocytic sarcoma 

13 (25 %) 9 (18%) 

1 (2%) 

9 (19%) 

1 (2%) 

1 (2%) 

9 08%) 

5 00%) 

Mast cell tumor NOS 1 (2%) 

Osteosarcoma, metastatic, uncertain 

primary site 1 (2%) 

Mesentery (6) (2) (7) (2) 

Carcinoma, metastatic, kidney I (14%) 

Leiomyosarcoma, metastatic, stomach, 

glandular 

Fat, hemangioma 

Fat, sarcoma 1 (17%) 

(14%) 

I (50%) 

Pancreas 

Carcinoma, metastatic, kidney 

Leiomyosarcoma, metastatic, stomach, 

(52) (50) (48) 

1 (2%) 

(50) 

glandular 

Salivary glands 

Stomach, forestomach 

Mast cell tumor NOS 

(52) 

(52) 

(50) 

(50) 

1 (2%) 

1 

(48) 

(47) 

(2%) 

(49) 

(48) 

Leiomyosarcoma, metastatic, stomach, 

glandular 

Squamous cell papilloma 

Stomach, glandular 

Adenoma 

Carcinoid tumor malignant 

Carcinoma 

2 (4%) 

(52) 

2 (4%) 

(50) 

1 

2 

(47) 

1 

1 

1 

(2%) 

(4%) 

(2%) 

(2%) 

(2%) 

2 (4%) 

(48) 

1 (2%) 

Leiomyosarcoma 

Mast cell tumor NOS 1 (2%) 

1 (2%) 

Cardiovascular System 

Heart 

Osteosarcoma, metastatic, uncertain 

(52) (50) (48) (50) 

primary site I (2%) 
288 Dietary Restriction, NTP TR 460 

TABLE Flb 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study of Salicylazosulfapyridine: 

2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

:, Control 2,700mg/kg Control 2,700mg/kg 

2-Year and 3-Year Protocols (continued) -

Endocrine System 

Adrenal cortex 

Leiomyosarcoma, metastatic, stomach, 

glandular • 

Spindle ceil, adenoma 

Adrenal medulla 

Pheochromocytoma benign 

Isle~ pancreatic 

Adenoma 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

EPididymis 

Preputial gland 

Prostate 

Seminal vesicle 

Testes 

Interstitial cell, adenoma 

Hematopoietic System 

Bone marrow 

Mast cell tumor NOS 

Lymph node 

Mediastinal, carcinoma, metastatic, kidney 

Mediastinal, carcinoma, metastatic, 

harderian gland 

Renal, leiomyosarcoma, metastatic, 

stomach, glandular 

Lymph node, mandibular 

Carcinoma, metastatic, kidney 

Carcinoma, metastatic, harderian gland 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Hemangioma 

Hernangiosarcoma 

Histiocytic sarcoma 

Thymus 

Carcinoma, metastatic, harderian gland 

Carcinoma, metastatic, kidney 

(52) 

(52) 

(52) 

(48) 

(52) 

1 (2%) 

(52) 

(51) 

(52) 

(52) 

(52) 

(52) 

(4) 

1 (25%) 

(51) 

1 (2%) 

(52) 

(52) 

1 (2%) 

(51) 

1 (2%) 

(50) 

(50) 

(50) 

(47) 

(50) 

1 (2%) 

(5o) 

(50) 

(50) 

(50) 

(50) 

(50) 

1 (2%) 

(2) 

(50) 

(5o) 

1 (2%) 

(50) 

1 (2%) 

(46) 

(47) 

1 (2%) 

I (2%) 

(47) 

(48) 

1 (2%) 

(44) 

I (2%) 

(47) 

1 (2%) 

(48) 

(48) 

(48) 

(48) 

(48) 

1 (2%) 

(48) 

(15) 

1 (7%) 

1 (7%) 

(47) 

I (2%) 

(48) 

1 (2%) 

(48) 

I (2%) 

(41) 

I (2%) 

(50) 

(49) 

I (2%) 

(50) 

1 (2%) 

(48) 

1 (2%) 

(49) 

(so) 

(50) 

(50) .. 

(50) 

(50) 

(50) 

(17) 

(49) 

(50) 

(5O) 

I (2%) 

(42) 
289 

~- -:,~-.:~i~~ ~ ............ _ -. :~. ~ ~ : SS~ ~ ~ ........... ~ ~ 

I 

I 

i 

:iI 

:: 

Dietary Restriction, NTP TR 460 

TABLE ~lb 

Summary of the Incidence of Neoplasms fin Male Mice in the Dietary Restriction Study of Salficy~zosuifapyridine: 

2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 2,700 mg/kg Control 2,700 mg/kg 

2-Year and 3-Year Protocols (continued) 

Integumentary System 

Skin (52) (50) (48) (50) 

Subcutaneous tissue, carcinoma, metastatic, 

.harderian gland 1 (2%) 

Subcutaneous tissue, hemangioma , 1 (2%) 

Subcutaneous tissue, lipoma I (2%) 

Musculoskeletal System 

Bone (52) (50) (48) (50) 

Rib, carcinoma, metastatic, harderian gland 1 (2%) 

Skeletal muscle (2) (1) 

Carcinoma, metastatic, harderian gland 1 (50%) 

Hemangiosarcoma 1 (100%) 

Osteosarcoma, metastatic, uncertain 

primary site 1 (50%) 

Nervous System 

None 

Respiratory System 

Lung (52) (50) (48) (50) 

Alveolar/bronchiolar adenoma 10 (19%) 3 (6%) 8 (17%) 5 (10%) 

Alveolar/bronchiolar adenoma, multiple 1 (2%) 2 (4%) 

Alveolar/bronchiolar carcinoma 2 (4%) 13 (27%) 6 (12%) 

Carcinoma, metastatic, harderian gland 2 (4%) 

Carcinoma, metastatic, kidney 1 (2%) 

Hepatocellular carcinoma, metastatic, liver 1 (2%) 2 (4%) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Osteosarcoma, metastatic, uncertain 

primary site 1 (2%) 

Mediastinum, carcinoma, metastatic, 

harderian gland 1 (2%) 

Nosc (52) (50) (48) (50) 

Glands, carcinoma, metastatic, harderian 

gland 1 (2%) 

Special Senses System 

Harderian gland (3) (3) (8) (1) 

Adenoma 2 (67%) 3 (1oo%) 6 (75%) 1 (lOO%) 

Carcinoma 1 (33%) 2 (25%) 

Zymbal's gland (1) 

Carcinoma 1 (lOO%) 
290 Dietary Restriction, NTP TR 460 

TABIJ~ Flb 

Sum~nmv of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study of Salicylazosulfapyridine: " ' 

2-Year and 3-Year Restricted Feed Protocols (continued) : 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehide : 

~ Control 2,700 mg/kg Control 2,700 mg/kg 

2-Year and 3-Year Protocols (continued) 

Urinary System 

Kidney (52) (50) (48) (50) 

Leiomyosarcoma, metastatic, stomach, 

glandular 1 (2%) 

Cortex, adenoma 1 (2%) 

Cortex, carcinoma 1 (2%) 1 (2%) 1 (2%) 

Cortex, carcinoma, metastatic, harderian 

gland 1 (2%) 

Urinary bladder (52) (50) (48) (50) 

Systemic Lesions 

Multiple organs c (52) (50) (48) (50) 

Histiocytic sarcoma I (2%) 1 (2%) 

Lymphoma malignant 6 (12%) 3 (6%) 5 (10%) 9 (18%) 
~ ~_~ ~r~'----- 7" ~ ~-- ~ ..... ~-:---7~-----~---~ .......... 

l 

Dietary ]Restriction, NTP TR 46~ 29~ 

T~L~ ~b 

Sunam~ry o~ the ]Incidence o~. Heop~asans ~n M~e 1Mice .in ~the Dietary Restriction Study o~ Sa~cy~azosu~f~pyri~ine: 

2-Year ~r~d 3-Year Restricted Feed Protocols (continued) 

I 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 2,700 mg/kg Control 2,7~10 mg/kg 

2. ~" ear and 3-3rear Protocols (continued) 

Neoplasm Summary 

Total animals with primary neoplasms d 

15-Month interim evaluation 2 

2-Year and 3-year protocols 37 20 42 34 

Total primary neoplasms .I 15-Month interim evaluation 2 

2-Year and 3-year protocols 49 28 81 52 

Total animals with benign neoplasms 

15-Month interim evaluation 1 I 

2-Year and 3-year protocols 26 16 27 25 

Total benign neoplasms 

15-Month interim evaluation 1 

2-Year and 3-year protocols 29 18 35 31 

Total animals with malignant neoplasms 

15-Month interim evaluation 1 I~ 

2-Year and 3-year protocols 18 6 36 19 

Total malignant neoplasms 

15-Month interim evaluation 1 

2-Year and 3-year protocols 20 6 46 21 

Total animals with metastatic neoplasms 

2-Year and 3-year protocols 4 7 

Total metastatic neoplasms 

2-Year and 3-year protocols 16 18 

Total animals with malignant neoplasms 

of uncertain primary site 

2-Year protocol 1 

Total animals with uncertain neoplasmsbenign or malignant 

2-Year protocol 

Total uncertain neoplasms 

2-Year protocol 

a Number of animals examined microscopically at the site and the number of animals with neoplasm ~ I b Three animals that died in dosing accidents before the interim evaluation were included in the interim evaluation data 

c Number of animals with any tissue examined microscopically 

d Primary neoplasms: all neoplasms except metastatic neoplasms 

~ I 
292 Dietary Restriction, NTP TR 460 

TABLE l~2a 

Statistic~d Analysis of Primary Neoplasms in Male Mice in the Dietary Restriction Study of Salicylazosulfapyridine: -

Ad Libimm Feeding and Weight-Matched Controls Protocols 

2,700 mg/kg x 2,700 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Harderian Gland: Adenoma or Carcinoma 

Overall rate a 3/50 (6%) 2/5O (4%) 2150 (4%) 2/50 (4%) 

Adjusted rate b 7.5% 4.3% 

Terminal ratec 3/40 (8%) 2/46 (4%) 

First incidence (days) 728 (T) 728 (T) 

Life table test a P=0.436N 

Logistic regression test a P=0.436N 

Fisher exact test d P=0.500N 

Liver: Hepatocellular Adenoma 

Overall rate 13/50 (26%) 42/50 (84%) 8/50 (16%) 42/50 (84%) 

Adjusted rate 32.5 % 87.5% 17.8% 87.5% 

Terminal rate 13/40 (33 %) 40/46 (87%) 8/45 (18%) 40/46 (87%) 

First incidence (days) 728 (T) 497 728 (T) 497 

Life.table test P<0.001 P<0.001 . 

Logistic regression test P<0.001 P<0.001 

Fisher exact test P<0.001 P<O.001 

Liver: Hepatocellular Carcinoma 

Overall rate 13/.50 (26%) 8/50 (16%) 6/50 (12%) 8/50 (16%) 

Adjusted rate 29.2% 17.4% 12.9% 17.4% 

8/46 (17%) 5/45 (I 1%) 8/46 (17%) Terminal rate 9/40 (23%) 

First incidence (days) . 

420 728 (T) 574 728 (T) 

Lifd table test P=0.106N P=0.401 

Logistic regression test P=0.159N P=0.378 

Fisher exact test P=0.163N P=0.387 

Liver: Hepatocellular Adenoma or Carcinoma 

24/50 (48%) 44/50 (88%) 14/50 (28%) " 44/50 (88%) Overall rate 

Adjusted rate 54.3% 91.7% 30.3% 91.7% 

42/46 (91%) 13/45 (29%) 42/46 (91%) Terminal rate 20/40 (50%) 

First incidence (days) 420 497 574 497 

P=O.O01 . P<0.001 Life table test 

P<O.O01 P<0.001 Logistic regression test 

P<O.O01 P<0.001 Fisher exact test 

Lung: Alveolar/bronchiolar Adenoma 

Overall rate 11/50 (22%) 6/50 (12%) 6/50 (12%) 6/50 (12%) 

Adjusted rate 27.5% 13.0% 12.6% 13.0% 

4/45 (9%) 6/46 (13%) Terminal rate 11/40 (28%) 6/46 (13%) 

First incidence (days) , 728 (T) 728 (T) 574 728 (T) 

P=0.081N P=O.60~N Life table test 

P=0.081N P=0.618 Logistic regression test 

P=0.143N P=O.620N Fisher exact test 
:=~._ : ~--~:~:~ ~-- "--'~------~.~__~:_.~.~_~,~. ~ _ ~ ~__ ..... - ...... . . . . . . . . . . . ~ . . . . 

i 

i 

l~ie~-y I~est~c~ion~ ~ ~ 4(~ 293 

TA~L~ ~2~ 

~t~s~ic~ An~dys~s of ~Pr'hn~ry Neoplasms in 1V~e I~'~ice in ~he l~ietary l~estricfion $~udy of S~icy~osu~f~pyr~d~ne: [~ 

Ad Libi~tm I~eeding ~n~ Weight-I~,SI~tchet~ Controls ]Protocols (continued) 

I 

I 

, 

Ad Libitum2,700 mg/kg × Ad Libitum- Weight-Matched 

2,7~ mg/kg × 

Weight-Matched i  ̧ -

Fed Control Fed Control Control Control 

I i 

Lung: Alveolar/bronchiolar Carcinoma 

Overall rate 3/50 (6%) 5/50 (10%) 4/50 (8 %) 5/50 (10%) 

Adjusted rate 7.3 % 10.9 % 8.9 % 10.9 % 

Terminal rate 2/40 (5%) 5/46 (11%) 4/45 (9%) 5/46 (11%) 

First incidence (days) 689 728 (T) 728 (T) 728 (T) [ Life table test P=0.434 P=0.514 i Logistic regression test 

Fisher exact test 

P=0.392 

P=0.357 

P=0.514 

P=0.500 [ 

I Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 14/50 (28%) 11/50 (22%) 9•50 (18%) 11/50 (22%) 

Adjusted rate 34.1% 23.9% 19.0% 23.9% 

Terminal rate 13/40 (33%) 11/46 (24%) 7/45 (16%) 11/46 (24%) 

First incidence (days) 689 728 (T) 574 728 (T) 

Life table test P=0.192N P=0.419 

Logistic regression test P=0.218N P=0.391 

Fisher exact test P=0.322N P=0.402 

Stomach (FForestomach): Squamous Cell Papilloma 

Overall rate 3•50 (6%) 0•50 (0%) 5•50 (10%) 0•50 (0%) 

Adjusted rate 7.5% 0.0% 10.8% 0.0% 

Terminal rate 3/40 (8%) 0/46 (0%) 4/45 (9%) 0/46 (0%) 

First incidence (days) 728 (T) .-Y 610 -

Life table test P=0.098N P=0,033N 

Logistic regression test P = 0.098,N P = 0.033N 

Fisher exact test P = 0.121N P = 0.028N 

All Organs: Hemangiosarcoma 

Overall rate 3•50 (6%) 1/50 (2%) 2150 (4%) 1/50 (2%) 

Adjusted rate 7.5 % 2.2 % 

Terminal rate 3/40 (8%) 1/46 (2%) -, 

First incidence (days) 728 (T) 728 (T) 

Life table test P=0,257N 

Logistic regression test P=0.257N 

Fisher exact test P=0.309N 

~ I All Organs: Hemangioma or Hemangiosarcoma 

Overall rate 4•50 (8%) 2/50 (4%) 2/50 (4%) 2/50 (4%) 

Adjusted rate 10.0% 4.3 % 

Terminal rate 4/40 (10%) 2/46 (4%) 

First incidence (days) 728 (T) 728 (T) 

Life table test P=0.275N 

Logistic regression test P = 0.275N 

Fisher exact test P=0.339N 
294 Dietary Restriction, NTP TR 460 

TABL£ F2a 

Statistical Analysis of Primary Neoplasms in Male Mice in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

2,700 mg/kg x 2,700 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

All Organs: Malignant Lymphoma (Lymphocytic or Unspecified) 

Overall rate 5/50 (10%) 2/50 (4%) 2150 (4%) 2/50 (4%) 

Adjnsted rate 12.1% 4.3 % 

Terminal rate 4140 (10%) 2146 (4%) 

First incidence (days) 666 728 (T) 

Life table test P=0.169N 

Logistic regression test P=0.198N 

Fisher exact test P=0.218N 

All Organs: Benign Neoplasms 

Overall rate 26/50 (52%) • 43/50 (86%) • 19/50 (38%) 43/50 (86%) 

Adjusted rate • 65.0% 89.6% 39,5% 89,6% 

Terminal rate 26/40 (65%) 41/46 (89%) 16/45 (36%) 41/46 (89%) 

First incidence (days) 728 (T) 497 574 497 

Life table test P=0.003 P<0.001 

Logistic regression test P < 0.001 P < 0.001 

Fisher exact test P < 0.001 P < 0.001 

All Org~ms: Malignant Neoplasms 

Overall rate 25/50 (50%) 15/50 (30%) 16/50 (32%) ' 15/50 (30%) 

Adjusted :rate' 53.2% 32.6% 33.3 % 32.6% 

Terminal rate 18/40 (45%) 15/46 (33%) 13/45 (29%) 15/46 (33%) 

First incidence (days) 420 728 (T) 574 728 (T) 

Life table test P=0~014N P=0.476N 

Logistic regression test , P=0.028N P=0.515N 

Fisher exact test P--0.033N Pf0.500N 

All Organs: Benign or Malignant Neoplasms 

Overall rate 40/50 (80%) 45/50 (90%) 30/50 (60%) 45/50 (90%) 

Adjusted rate 85. 1% 93.8 % 61.2 % 93.8 % 

Terminal rate 33140 (83%) 43/46 (93%) 26•45 (58%) 43146 (93%) 

First incidence (days) 420 497 574 497 

Life table test P--0.543N P=0.003 

Logistic regression test, P--0.137 P < 0.001 

Fisher exact test P=0,131 P<0.001 

('DTerm~d sacrifice 

a Number of neoplasm-bearing animals/number Of animals examined. Denominator is number of animals examined microscopically for liver, lung, and 

stomach; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

¢ Observed incidence at terminal kill 

d Beneath the "dosed group incidence are the P values corresponding to pailrwise comparisons between the ad libitum-fed or weight-matched controls and the 

dosed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in the 

dosed group is indicated by N. 

• Not applicable; no neoplasms in animal group 
~ ~ -- ;: = --= ~.~-~ : ~ ~: 77===7. 7:~'--~7~ .... ~ ........ . . 

Dietary ]Restriction, I~ T~ ~ 2~ 

T~ F~ 

} S~s~c~ 

~-~ ~ 

An~ys~s o~ ~ 

3-~ ~r~c~e~ 

N~#~ ~ 

~ee~ ~o~o~ 

MaZe ~ce ~ the ~e~ 

I 

2-Year ~estricted Feed 

I 

I Vehicle I 

I Control 2,700 m~cg 

I 

I

! 

~arder~an Gland: Adenoma 

Overall rate a 2/52 (4%) 3/50 (6%) 

Adjusted rate b 4.5% 8.6% 

Terminal rate c 1/42 (2%) 3/44 (7%) 

i First incidence (days) 660 730 (T) 

Life table test a P=0.511 

Logistic regression test d P =0.479 

Fisher exact test d P=0.481 

]~arderian Gland: Adenoma or Carcinoma 

Overall rate 3/52 (6%) 3/50 (6%) !~ I Adjusted rate 6.7% 8.6% 

Terminal rate 1/42 (2%) 3144 (7%) 

First incidence (days) 660 730 (T) 

Life table test P=0.649N 

Logistic regression test P=0.641 

Fisher exact test P=0.642 

Liver: Hepatocellular Adenoma 

Overall rate 13/52 (25%) 9/50 (18%) 

Adjusted rate 34.6% 22.7% 

Terminal rate 10/42 (24%) 8/44 (18%) 

First incidence (days) 616 648 

Life table test P=0.217N 

Logistic regression test P=0.274N 

Fisher exact test P=0.269N 

Liver: Hepatocellular Carcinoma 

Overall rate 7•52 (13%) 1/50 (2%) 

Adjusted rate 14.9% 2.3% 

Terminal rate 4•42 (10%) 1/44 (2%) 

First incidence (days) 495 729 (T) 

Life table test P=0.036N 

Logistic regression test P=0.025N 

~i~[ Fisher exact test P=0.034N 

Liver: ~Iepatocelinlar Adenoma or Carcinoma 

Overall rate 18/52 (35%) 9/50 (18%) 

Adjusted rate 42.2 % 22.7 % 

Terminal rate 12/42 (29%) 8/44 (18%) 

First incidence (days) 495 648 

Life table test P=0.040N 

Logistic regression test P=0.044N 

Fisher exact test P=0.046N 

~o~ S~u~y o~ ~cy~u~ap~e: .. 

3-Year ~estr~cted Feed 

Vehicle 

Control 2,7~ mg/~ 

6/48 (13%) 1/50 (2%) 

28.6% 2.8% 

5•20 (25%) 0•34 (0%) 

1,079 1,076 

P=0.010N 

P=0.008N 

P=0.050N 

8/48 (17%) 1/50 (2%) 

32.3% 2.8% 

5•20 (25%) 0/34 (0%) 

750 1,076 

P=0.003N 

P=0.007N 

Pffi0.013N 

10/48 (21%) 14/50 (28%) 

35.0% 38.6% 

5•20 (25%) 12/34 (35%) ~ 

555 933 

P=0.468N 

P=0.398 

P=0.278 

16/48 (33%) 6•50 (12%) 

42.2% 15.5% 

3•20 (15%) 3•34 (9%) 

445 726 

P=0.003N 

P=0.020N 

P=0.011N 

-21/48 (44%) 18/50 (36%) 

55.5 % 47.0% 

6/20 (30%) 14/34 (41%) 

445 726 

P=0.042N 

P=0.292N 

P=0.282N 
-- 

296 Diatary Restriction, NTP TR 460 

TABLE F2b 

Statistical Analysis of Primary Neoplasms in Male Mice in the Dietary Restriction Study of Salicylazosulfapyridine 

2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 2,700 mg/kg Control 2,700 mg/kg 

Lung: .Alveolar/bronchiolar Adenoma 

Ovettall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fishier exact test 

Lungi" Alveolar/brofichiolar Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) ~ 

Life table test 

Logistic regression test 

Fisher exact test 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rote 

Adjusted mte 

Termi0al rate 

First inci&mqe. (days) 

Life table test , .5"-

Logistic regressioh test 

Fisher exact test 

All Organs: Hemangiosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Hemangioma or Hemangiosarcoma 

Overall rate 

Adjusted rate 

_Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test , -

11/52 (21%) 

53.2% 

9142 (21%) 

717 

2/52 (4%) 

13/52 (25%) 

63.4% 

10/42 (24%) 

661 

1/52 (2%) 

, 1/52 (2%) 

3/50 (6%) 

7.9% 

3/44 (7%) 

729 if) 

P=0.020N 

P=0.023N 

P=0.025N 

0/50 (0%) 

3/50 (6%) 

7.9% 

3/44 (7%) 

729 (T) 

P=0.007N 

P=0.009N • 

P=0.008N 

0/50 (0%) 

0/50 (0%) 

8/48 (17%) 

33.1% 

5/20 (25%) 

862 

13/48 (27%) 

38.4% 

3•20 (15%) 

718 

18/48 (38 %) 

55.2% 

7/20 (35%) 

• 718 -, 

3/48 (6%) 

12.0% 

2/20 00%) 

569 

4/48 (8 % ) 

16.8% 

3/20 (15%) 

569 

7/50 (14%) 

18.6% 

5/34 (15%) 

719" 

P=0.171N 

P=0.335N 

P=0,465N 

6/50 (12%) 

17.1% 

5/34 (15%) 

1,076 

P=0.010N 

P=0.049N 

P=0.051N 

12/50 (24%) 

31.8% 

9/34 (26%) 

719 

P=0.011N 

P=0.075 N 

P=0.109N 

0/50 (o%i 

0.0% 

0/34 (0 %) 

P=0.066N 

P~0.123N 

P=0.114N 

2/50 (4%) 

5.3% 

1/34 (3%) 

933 

P=0.167N 

P=0.308N 

P=0.319N 
i 

ii 

I 

;;~ 7 : - - ~.~.~S~ ~--- - - : 7: ........ ?~S:__--~,.~: - : ~ i, :& !-~% ...... ~ ............... ~ ~ ~,, i,i. . . . . . , 

Dietary Restriction, NTP TEL 4~ 2~7 

;. 

TABLE ~b 

Stat~st~ca~ Analysis o~' Pr'hnary Neop~asn~s ~n ~e ~ce 

2-~ and 3-Y~ R~ricte~ Feed ~ot~o~ .(con~u~) 

~n the D~e~ ~r~ct~on St~y o~ S~cy~ap~e 

2-Year Restricted Feed 3-Year II~estricted Feed 

i 

Vehicle 

Control 2,70~ mg/kg 

Vehicle 

Control 2,7~0 mg/kg 

All Organs: Unspecified Malignant Lymphoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Benign Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression t~st 

Fisher exact test 

All Organs: Malignant Neoplasms ~ 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

6/52 (12%) 3/50 (6%) 5/48 (10%) 9/50 (18%) 

13.4% 6.9% 20.1% 21.1% 

4/42 (10%) 1/44 (2%) 2/20 (10%) 3/34 (9%) 

647 214 921 314 

P=0.245N P=0.501 

P =0.183N P=0.201 

P=0.264N P=0.217 

26•52 (50%) 16150 (32%) 27/48 (56%) 25150 (50%) 

81. i % 40.1% 83.2% • 65.5% 

22•42 (52%) 15/44 (34%) 15/20 (75 %) 21134 (62%) 

616 648 555 719 

P=0.029N P=0.0ff7N 

P =0.049N P=0.110N 

P=0.050N P=0.338N 

19/52 (37%) 6/50 (12%) 36•48 (75%) 19/50 (38%) 

42.9% 13.5 % ' 79.2 % 42.7% 

11/42 (26%) .3/44 (7%) 11/20 (55%) 9/34 (26%) 

495 214 445 314 

P=0.006N P<0.001N 

P=0.002N P-~0.008N 

P = 0.004N P < 0.001N 

All Organs: Benign or Malignant Neoplasms 

Overall rate 37•52 (71%) 20•50 (40%) 42148 (88%) 34•50 (68%) 

Adjusted rate 90.8 % 46.2 % 93.1% 77.1% 

Terminal rate 28/42 (67%) 16/44 (36%) 17/20 (85%) 24134 (71%) 

First incidence (days) 495 214 445 314 

Life table test P=0.002N P<0.001N 

Logistic regression test P=0.001N P=0.015N 

Fisher exact test P=0.O01N P=0.018N 

I 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for liver and lung; for 

other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intereurrent mortality ~: 

¢ Observed incidence at terminal kill 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the controls and that dosed group. The life table test 

regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in a dosed group is indicated by N. 

I 

• Not applicable; no neoplasms in animal group 
298 Dietary Restriction, NTP TR 460 

TABLE, F3a , 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols a 

DiSposition Summary 

Animals initially in study 

IS-Month interim evaluation 

Early deaths 

Accidental deaths 

Mofibtmd 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15.Month Interbn Evaluation 

Alimentary System 

Liver 

Clear cell focus 

Cytologic alterations 

Ensin0philie focus 

Hematopaietic cell proliferation 

Vacuolization cytoplasmic 

Salivary glands 

Vaanolization cytoplasmic 

Submandibular gland, depletion secretory 

Ston~ch, forestomach 

Hyperplasia 

Inflammation, subacute 

Stomach, glandular 

Erosion 

Endocrine System 

Adrvnal cortex 

Hypertrophy 

Capsule, hype~plasia 

Parathyroid gland ........ 

Cyst 

Thyroid gland 

Follicle, degeneration 

Genital Sy~em 

Preputial gland 

Duet, cyst 

Ad LibitumFed Control 

60 

10 

(1o) 

1 (10%) 

10 0OO%) 

(I0) -> 

(lO) 

(Io) 

I (1o%) 

(I0) , 

1 (10%) 

(I0) 

(10) 

1 (10%) 

00) 

4 (4O%) 

Weight-Matched 

Control 

60 

10 

I 

3 

1 

45 

60 

(1o) 

(lo) 

(10) 

3 (30%) 

2 (20%) 

(10) 

(I0) 

1 (10%) 

(I0) 

(10) 

00) 

8 (80%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 

J 

2,700 mg/kg 

60 

10 

(io) 

10 (I00%) 

1 (10%) 

1 (10%) 

(10) 

1 (10%) 

1 (10%) 

(I0) 

(I0) 

(10) 

1 (10%) 

1 (10%) 

(10) 

I (10%) 

(10) 

1 (10%) 

00) 

7 (70%) 
77 - ~ ~ : . ~:~ ..... ~ ....... .... :~ = .............. ~=':-'~:7--~,=------':: =" ~ - - - ~ - ~ 

I 

I Dietary Restriction, NT~ T~ ~6~ 2~ 

[ -" 

TABLE ~3a 

i 

' 

Summary of the Incidence of Nonneop~astic Lesions in Male Mice in the Dietary ]Restriction Study 

of SaHcyKazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls ]Protocols (continued) 

,

[ 

Ad LibitumFed Control 

Weight-Matched 

Control 2,700 mg/kg 

•

i 

t 

} 

I 

! 

, 

! 

: i 

i 

i 

[ i: [

i 

IS-Month £nterim Evalua~on (continued) 

Hematopoietic System 

Lymph node, mesenteric (10) 

Angiectasis 

Spleen ( I O) 

Hematopoietic cell proliferation 2 (20%) 

Pigmentation,hemosiderin 

Thymus (9) 

Cyst 

(i0) 

(IO) 

(Io) 

(1o) 

1 (lO%) 

(1o) 

lo (1oo%) 

10 (100%) 

(io) 

1 00%) 

~J 

Respiratory System 

Lung 

Alveolus, infiltration cellular, histioeyte 

Mediastinum, hemorrhage 

Nose 

Olfactory epithelium, degeneration 

(10) 

1 (10%) 

(10) 

1 (10%) 

(io) 

i (lO%) 

1 (10%) 

(I0) 

(10) 

1 (1o%) 

(1o) 

2 (20%) 

~: 

.S 

Urinary System 

Kidney 

Mineralization 

Renal tubule, casts 

Renal tubule, regeneration 

(10) 

8 (80%) 

1 (10%) 

7 (70%) 

(10) 

6 (60%) 

2 (20%) 

5 (50%) 

(10) 

2 (20%) 

1 (10%) 

4 (~%)r ' 

= 

= 

~ 

I 

I 

I 

[ 

[ 

I 

I

i 

{ 

Systems Examined With No Lesions Observed 

Cardiovascular System 

'General Body System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

2-%ear Study 

~ 

! 

; 

[ 

Esophagus 

Hemorrhage 

Intestine large, cecum 

Intestine small, duodenum 

Dila~tion 

Intestine stall, jejunum 

Hyperplasia, glandular 

Intestine small, ileum 

Inflammation, chronic 

Peyer's patch, hyperplasia, lymphoid 

(5o} 

t C2%) 

(47) 

t (2%) 

(47) 

(47) 

(47) 

(so} 

1 

(so) 

(so) 

(50) 

1 

(so) 

(2%) 

(2%) 

(2%) 

(50) 

(SO) 

(SO) 

I 

(so) 

(so) 

1 

(2%) 

(2%) 

__ = ̧: ̧ 

]~.:; : I 

~ : ! 
300 Dietary Restriction, NIT TR 460 

TABLE F3a 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine:, Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) ~ 

Ad Libitum- Weight-Matched 

Fed Control Control 2,7oo mg/kg 

2-Year Study (continued) 

Alimentary System (continued) 

Liver 

Angiectasis • 

Basophilic focus 

Clear cell focus 

Clear cell fo.cus~ multiple 

Eosinophilic focus 

Eosinophilic focus, multiple 

Faw/change, focal 

Hemorrhage 

Inflammation, chronic 

Inflammation, subacute 

Mineralization " ' 

Mixed cell focus 

Necrosis 

Pigmentation, bile 

Vaeuolization cytoplasmic 

Bile duct, cyst 

Mesentery 

Fat, inflammation, subacute 

Fat, necrosis 

Pancreas. . . .~ 

Necrosis 

Acinus, atrophy 

Acinus, depletion secretory 

Stomach, forestomach 

Diverticulum 

Edema 

Hyperplasia 

Inflammation, subacute 

Ulcer 

stomach, glandular 

Erosion 

Hyperplasia 

Inflammation, subacute 

Mineralization 

Pigmentation 

Ulcer 

Cardiovascular System 

Heart 

Embolus 

Mineralization 

(50) 

1 (2%) 

2 (4%) 

5 (lO%) 

1 (2%) 

1 (2%) 

1 (2%) 

3 (6%) 

2 (4%) 

2 (4%) 

3 (6%) 

1 (2%) 

3 (6%) 

2 (4%) 

(4) 

3 (75%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

18 (36%) 

4 (8%) 

3 (6%) 

(50) 

I (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

(5o) 

1 (2%) 

i (2%) 

(5o) 

i (2%) 

2 (4%) 

1 (2%) 

1 (2%) 

I (2%) " 

2 (4%) 

7 (14%) 

(2) 

1 (50%) 

I (50%) 

(50) 

I (2%) 

(5O) 

. 

I (2%) 

22 (44%) 

11 (22%) 

2 (4%) 

(50) 

I (2%) 

1 (2%) 

(5o) 

(50) 

8 06%) 

3 (6%) 

13 (26%) 

9 (18%) 

1 (2%) 

2 (4%) 

3 (6%) 

(1) 

I (I00%) 

(50) 

I (2%) 

(50) 

lO (20%) 

2 (4%) 

(50) 

I (2%) 

7 (14%) 

(50) 
--___ 

=~7~÷-'-~-'" ~'~7S-"L-S--~---~7~'~'~ - - ~ ~-2 ;;,~:q~:: ; ~ .... 7 ;----~,~-"--~ :~ -:--_ : ~ .... ,, _ _= 

D~e~ Restriction, NTF TR ~60 30~. 

TABLE ~ 

Sun~n~'y of the l[ncidence of Nonneop~s~ic Lesions ~n ]V~e 1V~ice in ~he ~ie~-y Res~ction Study 

o~ S~licy]~osu~f~pyr[dh~e: Ad Libitum ~ee~ng ~d We~gh~-M~che~ Con~o~s Protocols (~on~ued) 

. 

I Ad Libitum- Weight-Matched 

Fed Control Control 2, 7~0 rag/kS 

i 

2- Irear Study (continued) 

I 

! 

Endocrine System 

Adrenal cortex 

Cyst 

Embolus 

Hyperplasia 

Hypertrophy 

Vacuolization cytoplasmic 

Capsule, hyperplasia 

Adrenal medulla 

Embolus 

Islets, pancreatic 

Embolus 

Hyperplasia 

parathyroid gland 

Cyst 

Pituitary gland 

Pars distalis, cyst 

Thyroid gland 

Follicle, cyst 

Follicle, degeneration 

Follicular cell, hyperplasia 

(50) 

1 

8 

11 

1 

I 

(50) 

1 

(50) 

1 

2 

(48) 

(44) 

(50) 

6 

3 

9 

(2%) 

(16%) 

(22%) 

(2%) 

(2%) 

(2%) 

(2%) 

(4%) 

(12%) 

(6%) 

(18%) 

(50) 

1 (2%) 

8 (16%) 

(50) 

(48) 

(46) 

4 (9%) 

(50) 

4 (8%) 

3 (6%) 

1 (2%) 

(50) 

6 

12 

4 

(50) 

(50) 

3 

(49) 

. .1 

(47) 

(50) 

9 

3 

2 

(12%) 

(24%) 

(8%) 

(6%) 

(2%) 

(18%) 

(6%) 

(4%) 

i 

i 

i ~ 

General Body System 

None 

Genital System 

Coagulating gland 

Dilatation 

Inflammation, chronic 

Epididymis 

Congestion 

Granuloma sperm 

Inflammation, chronic 

Preputial gland 

Congestion 

Inflammation, subacute 

Duct, cyst 

Prostate 

Cyst 

Hyperplasia 

Inflammation, suppurative 

Seminal vesicle 

Atrophy 

Dilatation 

Inflammation, chronic 

Testes 

Atrophy 

Mineralization 

Interstitial cell, hyperplasia 

(5) 

5 

(50) 

1 

1 

1 

(49) 

1 

2 

45 

(50) 

1 

1 

1 

(50) 

2 

14 

1 

(50) 

1 

(IOO%) 

(2%) 

(2%) 

(2%) 

(2%) 

(4%) 

(92%) 

(2%) 

(2%) 

(2%) 

(4%) 

(28%) 

(2%) 

(2%) 

(50) 

(50) 

42. (84%) 

(50) 

(50) 

1 (2%) 

(50) 

(1) 

1 0o0%) 

(50) 

(50) 

38 (76%) 

(50) 

(50) 

2 (4%) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 
302 Dietary Restriction, NTP TR 460 

TABLE F3a 

Smnmary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched ContrOls Protocols (continued) ,, 

2-Year Study (continued) 

Hematopoietic System 

Bone marrow 

Atrophy 

Myeloid cell, hyperplasia 

Lymph node 

Inguinal, hyperplasia, lymphoid 

Inguinal, infiltration cellular, mast cell 

Mcdiastinal, hypcrplasia, lymphoid 

Renal, hyperplasia, lymphoid 

Lymph node, mandibular 

Hypcrplasia, lymphoid 

Lymph node, mesentcric 

Angiectasis 

Hypcrplasia, lymphoid 

Spleen 

Atrophy 

Congestion , 

Hcmatopoietic cell proliferation 

Hyperplasia, lymphoid 

Pigmentation, hemosiderin 

Thymus 

" Atrophy 

Cyst 

Mineralization 

Thymocyv=, necrosis 

Integumentary System 

Skin 

Hyperkeratosis 

Subcutaneous tissue, edema 

Subcutaneous tissue, inflammation, chrome 

Subcutaneous tissue, metaplasia, osseous 

Subcutaneous tissue, mineralization 

Musculoskeletal System 

Bone 

Cranium, hyperostosis 

Nervous System 

Brain 

Compression 

Embolus 

Hydrocephalus 

Pigmentation, hemosiderin 

Vacuolization cytoplasmie 

Ad LibitumFed Control 

(5o) 

2 (4%) 

(3) 

1 (33%) 

(50) 

(50) 

17 (34%) 

4 (8%) 

(50) 

5 (lO%) 

I (2%) 

H (22%) 

5 (1o%) 

2 (4%) 

(45) 

3 (7%) 

2 (4%) 

I (2%) 

(50) 

1 (2%) 

1 (2%) 

(50) 

2 (4%) 

(50) 

i (2%) 

1 (2%) 

Weight-Matched 

Control, . : 

(50) 

2 (4%) 

(i) 

(49) 

I (2%) 

(48) 

23 (48%) 

3 (6%) 

(50) 

3 (6%) 

8 (16%) 

I (2%) 

(46) 

I (2%) 

I (2%) 

(50) 

1 (2%) 

1 (2%) 

(50) 

i (2%) 

(5o) 

I (2%) 

! (2%) 

2,700 mg/kg 

(50) 

(4) 

I (25%) 

I (25%) 

I (25%) 

(49) 

I (2%) 

(50) 

12 (24%) 

3 (6%) 

(50) 

• i (2%) 

13 (26%) 

7 (14%) 

47 (94%) 

(46) 

1 (2%) 

1 (2%) 

I (2%) 

(50) 

I (2%) 

(50) 

(~o) 

1 (2%) 
. ~ 2 ~ , ~ ~ : ~ _ ~ 

~ ..... . :~7 ..... ~'~C'~"~ ~ : ~ i.i .i . . . . 7-~ ~:~ " ~ ....... ~ ................. ..-- ~-~2"; ~ ~-,.--.~:?SS~.~.~.~ ........... , 

~ie~'yj lI~es~ict~on, ~TII~ ~I) 303

T&~L~ F3a 

S~ary o~ the Incipience o~ Honneop~stic Lesions ~n ~a~e Mice In the Dietary Restriction St~ly 

o~ Sa~icy~s~apyrt~ine: Ad Libitura ]~ee~ng and Weight-I~atche~ Controls ]?r-oteco~s (continued) 

Ad Libitum- Weight-I~5Iatched 

I Fed Control Control 2,700mg/~g i :  ̧ 

I 

I 

~ ??l 2oYear S~dy (continued) 

] Respiratory System 

I Lung (50) (50) (50) 

l Congestion ! (2%) 

[ • 

~ i 

Hyperplasia, maerophage 

Alveolar epithelium, hyperplasia 4 (8%) 

1 (2%) 

3 (6%) 

! (2%) 

8 (16%) i -

~ ~ [ .[ Alveolus, foreign body 

Alveolus, hemorrhage 

1 

1 

(2%) 

(2%) 

! (2%) 

I (2%) 

~ i Alveolus, infdtration cellular, histioeyte 3 (6%) 1 (2%) 

,I Mediastinu.m,hemorrhage 1 (2%) 

[ Nose (50) (50) (50) 

: ~ } Glands, cyst 1 (2%) I (2%) 

i 

Speciad Senses System, 

None 

Urinary System 

Kidney 

Bacterium 

(50) 

I (2%) 

(50) (50) 

Fibrosis 

Glomeruloselerosis 

Infarct 

Infdtration cellular, lymphocyte 

1 (2%) 

3 (6%) 

I (2%) 

1 (2%) (2%) 

Inflammation,subacute 

Metaplasia, osseous. 

Mineralization 

Cortex, cyst 

2 (4%) 

37 (74%) 

5 (10%) 

44 (88%) 

4 (8%) 

2 

33 

1 

(4%) 

(66%) 

(2%) 

i 

Cortex, medulla, inflammation, suppurative I (2%) 

Pelvis, necrosis ! (2%) 

Renal tubule, casts 13 (26%) 7 (14%) 3 (6%) 

Renal tubule, degeneration 

Renal tubule, regeneration 

3 (6%) 

38 (76%) 41 (82%) 

1 

24 

(2%) 

(48%) 

Transitional epithelium, hyperplasia ! (2%) 

Urinary bladder 

Dilatation 

Inflammation, subacute 

Transitional epithelium, hyperplasia 

(50) 

i (2%) 

! (2%) 

1 (2%) 

(50) (50) 

1 (2%) 

i 

i 
304 Dietary Restriction, NTP TR 460 

TABLE F3b 

Smnmary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 3-Year Restricted Feed Protocols a 

Disposition Summary 

Animals initially in study 

lS-Month interim evaluation 

Accidental deaths b 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

Alimentary System 

Intestine large, colon 

•Inflammation, chronic 

Liver 

• Basophilic focus 

Cytologic alterations 

Vacuoliz~ition cytoplasmic 

Mesentery 

Fat, inflammation, granulomatous 

Salivary glands 

Submandibular gland, depletion secretory 

Submandibular gland, vacuolization 

cytoplasmic 

Stomach, forestomach 

Hyperplasia 

Inflammation, subacute 

Stomach, glandular 

Mineralization 

Endocrine System 

Adrenal cortex 

Hypertrophy 

Thyroid gland 

Follicle, cyst 

Follicle, degeneration 

Genital System 

Preputial gland 

Duct, cyst 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 2,700 mg/kg •Control 2,700 mg/kg 

62 60 48 50 .• . 

8 9 

2 1 

~ ~ 21 

4 1 7 

42 44 20 34 

62 60 48 50 

(lo) (1o) 

1 (10%) 

(lO) (10) 

1 (10%) 

6 (60%) 

3 (30%) 

(1) 

1 (100%) 

(10) (10) 

1 (10%) 3 (30%) 

4 (40%) 

(10) (10) 

4 (40%) 2 (20%) 

4 (4o%) 

(1o) (10) 

1 (10%) 

(1o) (lo) 

1 (1o%) • 

(1o) (lo) 

1 (10%) 

1 (10%) 

(10) (10) 

8 (80%) 4 (40%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 

b Three animals that died in dosing accidents before the interim evaluation were included in the interim evaluation data 
--..~ ~.~:--" __ : ~ ~==~:-~ :~ ---,~.__ ............. 

!

!

[

[ 

Dietary Restriction, NTP TR 460 305 

! 

i 

I 

TABLE F3b 

Summary of the Incidence of N0nneoplastic Lesions in Male Mice in the Dietary Restriction Study . 

of Salicylazosulfapyridine: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

! 

. 

i: ~ 

i 

Vehicle 

Control 2,700 mg/kg 

Vehicle 

Control 2,700 mg/kg 

i 15-Month ffnterim Evaluation (continued) 

Hematopoietic System 

Spleen 

Hematopoietic cell proliferation 

Pigmentation, hemosiderin 

(lO) (lo) 

9 

lo 

(90%) 

(lOO%) 

i 

I Integumentary System 

Skin 

Epidermis, hyperplasia 

(10) 

1 (10%) 

(lO) :~ 

: 

Respiratory System 

Lung 

Hemorrhage 

(10) (10) 

1 (10%) 

i 

i

i 

Urinary System 

Kidney 

Atrophy 

Metaplasia, osseous 

Mineralization 

Cortex, cyst 

Pelvis, inflammation, subacute 

Renal tubule, casts 

Renal tubule, regeneration 

Urinary bladder 

Inflammation, subacute 

Transitional epithelium, hyperplasia 

(10) 

1 (10%) 

5 (50%) 

2 (20%) 

4 (40%) 

(10) 

(10) 

1 

1 

1 

4 

2 

(10) 

1 

1 

(10%) 

(10%) 

(10%) 

(40%) 

(20%) 

(10%) 

(10%) 

i_ 

i 

Systems Examined With No Lesions Observed 

Cardiovascular System 

General Body System 

Musculoskeletal System 

Nervous System 

Special Senses System 

i 

2-Year and 3-Year Protocols 

Alimentary System 

Gallbladder 

Dilatation 

Pigmentation 

Intestine small, jejunum 

Dilatation 

Peyer's patch, hyperplasia, lymphoid 

(51) 

(51) 

(50) 

(50) 

1 (2%) 

2 (4%) 

(48) 

(47) 

1 (2%) 

(49) 

1 (2%) 

1 (2%) 

(46) 
306 .Dietary Restriction, .NTP TR 460 

TABLE F3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the DietaryRestriction Study 

of Salicylazosulfapyridine: 2-Year and 3-Year Restricted Feed Protocol (continued) 

2-Year Restricted Feed 

Vehicle 

' Control 2,700 mg/kg 

2- Yea,r and 3-Year Protocols (continued) 

Alimentary System (continued) 

Liver (52) (50) 

Angiectasis 1 (2%) 

Basophilic focus 2 (4%) 

, 

1 (2%) 

Clear cell focus 1 (2%) 1 (2%) 

Clear cell focus, multiple 

Congestion 

Cytologic alterations 2 (4%) 

Eosinophilic focus. 5 (10%) i (2%) 

Fibrosis 

Hematopoietic cell proliferation I (2%) 

Hemorrhage I (2%) 

Hepatodiaphragmatic nodule 

Infiltration cellular, lymphocyte 1 (2%) 

Inflammation, subacute I (2%) 

Mixed cell focus I (2%) 

Necrosis 2 (4%) 2 (4%) 

Pigmentation, bile 1 (2%) 

Pigmentation, hemosiderin 2 (4%) 

Thrombosis 

Vacuolization cytoplasmic 3 (6%) 

Bile duct, cyst 1 (2%) 

Mesentery (6) (2) 

Fat, hemorrhage 

Fat, mineralization 1 (17%) 

Fat, necrosis 4 (67%) 1 (50%) 

Pancreas (52) (50) 

Cytoplasmic alteration 2 (4%) 

•Inflammation, subacute. I (2%) 

Artery, inflammation, chronic active 

Duct, cyst I (2%) 

Salivary glands (52) (5o) 

Degeneration 

Stomach, forestomach (52) (50) 

Foreign body I (2%) 

Hyperplasia 27 (52%) 1o (20%) 

Inflammation, subacute 12 (23%) 4 (8%) 

Ulcer 2 (4%) l (2%) 

Stomach, glandular (52) (50) 

Cyst 

Erosion 1 (2%) 

Hyperplasia 

Metaplasia, hepatocyte 

Mineralization 

Necrosis 

Pigmentation 

Tongue 

Hyperplasia, squamous 

Tooth (1) 

Developmental malformation I OOo%) 

Inflammation, subacute 

3-Year Restricted Feed 

Vehicle 

Control 2,700 mg/kg 

(48) (50) 

1 (2%) 1 (2%) 

4 (8%) 1 (2%) 

2 (4%) 1 (2%) 

1 (2%) 

1 (2%) 

(6%) 

4 (8%) (14%) 

(2%) 

1 (2%) (2%) 

I (2%) 

(2%) 

1 (2%) 

4 (8%) 

I (2%) 

I (2%) 

I (2%) I (2%) 

l (2%) 2 (4%) 

(7) (2) 

1 (50%) 

4 (57%) 

(48) (50) 

I (2%) 2 0%) 

I (2%) 

1 (2%) 1 (2%) 

(48) (49) 

1 (2%) 

(47) (48) 

18 (38%) • 14 (29%) 

I (2%) 1 (2%) 

4 (9%) I (2%) 

(47) (48) 

2 (4%) I (2%) 

4 (9%) 1 (2%) 

I (2%) 

I (2%) 

2 (4%) 

1 (2%) 

1 (2%) 

(I) 

I (100%) 

(8) (3) 

8 (100%) 3 (100%) 

I (13%) 
:i~ : ~.~.~.A;!5~---~.7--- - - _ ....... ~ ..................... - - - - - 7---7~: ....... :~-- :..~. ---~ ~ ~ .......... 

'Dietzry Restriction, NT~' TR 4~0 307 

TABLE ]F'3b 

Summary of the Incidence of Nouneop]astic Lesions in Male Mice in the Dietary Restriction S~udy 

of.Sa~icyiazosuff'apyridine: 2-~fear and 3-Year l~.estricted Feed Frotoco~s (continued) I 

2-Year Restricted Feed 3-Year Restricted Feed 

,Vehicle Vehicle I 

Control 2,700 mg/kg Control 2,700 mg/kg 

2-Year and 3-Year Protocols (cominued) ! Cardiovascular System 

Blood vessel (52) (50) (48) (50) 

Polyarteritis ! (2%) 

Heart (52) (50) (48) (5O) 

Cardiomyopathy i (2%) ! Fibrosis I (2%) 

Mineralization 2 (4%) 

== 

Endocrine System 

Adrenal cortex (52) (50) (47) (50) 

Accessory adrenal cortical nodule 1 (2%) 1 (2%) 

Hyperplasia 5 (11%) 2 (4%) 

Hypertrophy 8 (15%) 5 (1o%) 9 (19%) 8 (16%) 

Spindle cell, hyperplasia l (2%) 2 (4%) 3 (6%) 

Adrenal medulla (52) (50) (47) (49) 

Hyperplasia 1 (2%) 1 (2%) 

Pigmentation I (2%) 

Islets, .pancreatic (52) (50) (48) (50) 

Hype@lasia 3 (6%) I (2%) 1 (2%) 

Parathyroid gland (48) (46) (47) (47) 

Cyst 2 (4%) 1 (2%) 

Infiltration cellular, lymphocyte 1 (2%) 

Pituitary gland (48) (47) (44) (48) 

Pars distalis, cyst I (2%) I (2%) 4 (9%) 

Pars distalis, hyperplasia 2 (4%) I (2%) 

Thyroid gland (52) (50) (47) (49) 

Follicle, cyst 2 (4%) 8 (17%) 4 (8%) 

Follicle, degeneration 5 (10%) 2 (4%) 13 (28%) 16 (33%) 

Follicular cell, hyperplasia 7 (15%) 

General Body System 

None 

Genital System 

Epididymis (52) (50) (48) (50) 

Granaloma sperm 1 (2%) 

Hemorrhage 1 (2%) 

Infiltration cellular, lymphocyte 1 (2%) 

Mineralization 1 (2%) 2 (4%) 

Penis (1) 

Inflammation, subacute 1 000%) 

Preputial gland (51) (50) (48) (50) 

Inflammation, subacute 1 (2%) 2 (4%) 21 (44%) 11 (22%) 

Necrosis 1 (2%) 

Duct, cyst 44 (86%) 20 (40%) 43 (90%) 32 (64%) 
308 Dietary Restriction, NTP TR 460 

TABLE: F3b 

.Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Salieylazosulfapyridine: 2-Year and 3,Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 2,700 mg/kg Control 2,700 mg/kg 

2-Year and 3-Year Protocols (continued) 

Genital System (continued) 

Prostate (52) (50) (48) (50) 

Inflanunation, subacute 1 (2%) 

Seminal vesicle (52) (50) (48) (50) 

Atrophy I (2%) 5 (10%) 

Dilatation 1 (2%) 9 (19%) 

Infiltration cellular, mast cell I (2%)- 

Testes ' (52) (50) (48) (5o) 

Atrophy 1 (2%) 1 (2%) 

Mineralization 2 (4%) 

Hematopoietic System 

Lymph node (4) (2) (15) (17) 

Axillary, hyperplasia, lymphoid I (6%) 

Axillary,, pigmentation, hemosiderin I (6%) 

Bronchial, hyperpla~ia, lymphoid . 1 (25%) 

Inguinal, hyperplasia, lymphoid • 5 (33%) 1 (6%)

• Inguinal, infiltration cellular, masi cell 1 (7%) 

Inguinal, ~pigmentation 4 (27%) 10 (59%) 

Mediastinal, hemorrhage 1 (7%) 

Modiastinai, hyperplasia, lymphoid 1 (6%) 

Lymph node, mandibular (51) (50) (47) (49) 

Pigmentation, hemosiderin 3 (6%) 

Lymph node, mesenteric (52) (50) (48) (50) 

Angiectasis 20 (38%) 2 (4%) 24 (50%) 14 (28%) 

Edema 1 (2%) 

Hemorrhage (2%) 

Hyperplasia, lymphoid 5 (lO%) 2 (4%) (6%)' 

Inflammation, subacute (2%) 

Necrosis 1 (2%) 

Spleen (52) (50) (48) (50) 

Angiectasis I (2%) I (2%i

Atrophy 2 (4%) 3 (6%) 2 (4%) 5 (1o%) 

Congestion I (2%) ~ (2%) 

Depletion cellular I (2%) 

Hematopoietic cell proliferation 6 (12%) 5 (10%) 12 (25%) 7 (14%) 

Hyperplasia, lymphoid 9 (17%) 3 (6%) I (2%) 1 (2%) 

Pigmentation, hemosiderin 39 (78%) 37 (74%) 

Capsule, fibrosis 1 (2%) 

Thymus (51) (46) (41) (42) 

Atrophy 2 (4%) 2 (4%) 4 (10%) 2 (5%) 

Cyst t-(2%) 2 (5%) 1 (2%) 

Mineralization 1 (2%) 

Necrosis I,(2%) 
:T~ ..~L ..... r~ ~ ~ . . . . . . . . ~ ................ ~ ~ ......... ....... :~ " ~ = ~ 1 ~ : :~ ~ ....... ~ ........ ~ ~ i ~I~ ~ ~111 , 

Dietary Restriction, NT]P TR 460 309 

TABLE F3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restrictlon Study i 

of Sanicylazosuffapyridine: 2-Year and 3-Year Restricted Feed ]Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 2,700 mg/kg Control 2,700 mg/kg I 

I 2-Year and 3-Year Protocols (continued) 

Integumentary System 

Skin (52) (50) (48) (50) 

Edema 1 (2%) 

Hemorrhage 1 (2%) 

Inflammation, subacute 2 (4%) 1 (2%) 1 (2%) 

:=-

i 

I Ulcer I (2%) 

Epidermis, cyst 1 (2%) 

~l 

Musculoskeletal System 

Bone (52) (50) (48) (50) 

Hyperostosis 1 (2%) 

Sternum, developmental malformation 1 (2%) 

Nervous System 

Brain (52) (50) (48) (50) 

Atrophy 1 (2%) 

Hemorrhage 1 (2%) 

Vacuolization cytoplasmic 6 (13%) 10 (20%) 

Respiratory System 

Lung (52) (50) (48) (50) 

Congestion 1 (2%) 

Hemorrhage 1 (2%) 3 (6%) 2 (4%) 

Infiltration cellular, histiocyte 1 (2%) 

Pigmentation 2 (4%) 

Alveolar epithelium, hyperplasia 3 (6%) 2 (4%) 3 (6%) 9 (18%) 

Nose (52) (50) (48) (50) 

Glands, cyst 1 (2%) 

Glands, mineralization 3 (6%) ~: Inflammation, subacute 3 (6%) 

Lumen, inflammation, suppurative 1 (2%) 

Respiratory epithelium, mineralization 1 (2%) 

! ~ ! Special Senses System ~ ,~ 

i 

Ear (1) (1) 

External ear, inflammation, granulomatous 1 (loo%) 

Eye (1) 

Inflammation, suppurative 1 (100%) 
3~[0 ~.- Dietary Restriction, NTP TR 460 

TA~IJ~ F3b 

Sunnnary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

Vehicle 

Control 2,700 mg/kg 

2-Year and 3-Year Protocols (continued) 

Urinary System 

Kidney (52) (50) 

Fibrnsis 

Glomemlosclerosis 

Infarct ~ 5 00%) 2 (4%) 

Infiltration cellular, lymphocyte 

Inflammation, chronic 1 (2 %) 

Inflammation, subacute 1 (2%) 

Inflammation, suppurative 1 (2%) 

Metapllasia, osseous 1 (2%) 

Mineralization 49 (94%) 38 (76%) 

Artery, inflammation, chronic active . ., 

Cortex, cyst 1 (2%) 4 (8%) 

Renal tubule, casts 13 (25%) 20 (40%) 

Renal Illbule, degeneration I (2 %) 

Renal tubule, dilation 

Renal tubule, pigmentation 

Renai tubule, regeneration 36 (69%) 23 (46%) 

Urinary bladder (52) (50) 

Inflammation, subacute 

Transiti~onai epithelium,, hyperplasia 

Transitional epithelium, pigmentation 

3-Year Restricted Feed 

Vehicle 

Control 2,700mg/kg 

(48) (50) 

1 (2%) 1 (2%) 

7 (15%) 4 (8%) 

1 (2%) 3 (6%) 

1 (2%) 

I (2%) 

1 (2%) 1 (2%) 

41 (85%) 41 (82%) 

i (2%) 

15 (31%) 5 (10%) 

29 (60%) 26 (52%) 

4 (8%) 7 (14%) 

1 (2%) 

I (2%) 

27 (56%) 31 (62%) 

(48) (50) 

I (2%) 

1 (2%) 1 (2%) 

1 (2%) 1 (2%) 
I 

. . . . . = .............. ~ : ~ ::~=~;~:~ :: ........ ~ ................. 

! 

3~.~. 

. . 

AI~I~I~NIIX~X ~ i 

SUMMARY ©~F L~ES~I©N5 i~IN MAIL~E MI~CI~ 

I[N TItlE II~I~ETA~Y ~,dEST~I~JICT]I©N STUDY 

&IF $C©~©LAI~I]INIB IBY~]X©IB~©I~4I]I~ T~ff~YI~AT~E 

TABLE G~a Summary of the Incidence of Neoplasms in Male Mic.e 

nn the Dietary Restriction Study of Scopolmnine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 312 

TABLE Glh Summary of the Incidence of Neoplasms in Male Mice " • 

m the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

2-Year and 3-Year Restricted Feed Protocols .............................. 316 

TAI~LI~ G~a Statistical Analysis of Pr~nary Neoplasms in Male Mice 

m the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 320 

T~LE G~h ~tatlst~cal Analysis of ~rimary Neoplasms in Male Mice 

in the Dietary Restriction Study of Scopolamine Hydrobrom~de Tr~hydrate: 

2-Year and 3-Year Restricted Feed Protocols .............................. 3~ 

TABLE G3a Summary of the Incidence of Nonneoplast~c Lesions in Male Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ....... ~ ........... 32~ ' 

TABLE G3b Summary of the Kncidence of Nonneoplastic Lesions in Marie Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromlde Tr~hydrate: 

2-Year and 3-Year Restricted Feed Protocols .............................. 334 
312 Dietary Restriction, NTP TR 460 

TABLE Gla 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight~Matche d Controls Protocols a 

Ad LibitumFed Control 

Disposition Summary 

Animals initially in study 70 b. 

15-Month inte~m evaluation I0 

Early deaths 

Accidental deaths 

Moribund . 4 

Natural deaths 6" 

Survivor,,; 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

Alimentary System . 

Liver 

Hepatocellular carcinoma 

Hepatucellular adenoma 

Hepatocellular adenoma, multiple . 

il0) 

1 (10%), 

1 (10%) 

Respiratory System " . 

Lung (10) ' 

Alveolar/bronclfiolar adenoma 2 (20 ~), 

. , 

Systems Examined With No Neoplasms Observed 

Cardiovascular System 

Endocrine System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Special Senses System 

Urinary System "% _~_-~ 

Weight-Matched 

Control 25 mg/kg 

60 

10 

1 

2 

6 

41 

60 

70 b 

10 

2 

7 

2 

39 

1 (io%) 

(io) 

1 (1o%) 

(io) 

1 (10%) 

(10) 

~ 00~) 

.. 
~.~ -- - - __ _ ~--~:~'~':'~:--~--~~~ ................. ~ ~ ........ ~:a --==7- ~ ........................ 

~ie~ry Restriction, NTIP TR ,~ 313 

T~L~ G~ .. ~ . 

S~ary of the ]Incidence off Neoplasms in I~Iede Mince ~n the ~Je~ ~tr~cfion .Study 

I o~ Sco~ne ~ydrobrom~de Tr~hydrate: Ad ~b~m Feeding and Weight-Watched Control ~ot~o~ (confinu~) 

I Ad Libitum- Weight-Matched i 

Fed Control Control 25 mg/kg 

i 

2.Year Study 

Alimentary System 

Intestine small, duodenum (50) (50) (50) ~ l Adenoma 1 (2%) 

Intestine small, jejunum (50) (50) (50) 

Intestine small, ileum (49) (50) (48) 

Liver (50) (50) (50) 

Hemangioma I (2%) 

Hemangiosarcoma, metastatic, skin I (2%) 

Hemangiosarcoma, metastatic, spleen 1 (2%) i :~ 

Hepatocellular carcinoma (12%) 4 (8%) 7 (14%) i: 

Hepatocellular carcinoma, multiple 1 (2%) i : Hepatocellular adenoma 

Hepatoeellular adenoma, multiple 

10 

16 

(2O%) 

(32%) 

4 (8%) 

1 (2%) 

8 (16%) : 

!, 

Hepatocholangiocarcinorna 

Histiocytie sarcoma 

Ito cell tumor NOS, multiple 

3 

1 

(6%) 

(2%) 

i (2%) 

1 (2%) 

} 

1 

i 

i 

Mesentery 

Histiocytic sarcoma 

Pancreas 

Stomach, forestomach 

Mast c¢11 tumor benign 

(4) 

(50) 

(50) 

(50) 

(50) 

I 

(1) 

(2%) 

(2) 

1 (50%) 

(5o) 

(5o) 

ii 

I 

i~ 

Squamous cell papilloma 

Tooth 

Odontuma 

' 

(14) 

1 (7%) 

2 (4%) 

(2) 

1 (50%) ! 

[ 

Cardiovascular System 

}: 

None J 

[ :[ Endocrine System 

Islets, pancreatic (50) (50) (50)

Adenoma 3 (6%) 1 (2%) 

Pituitary gland (48) (48) (44) I Pars distalis, adenoma 1 (2%) 

I 

Pars intermedia, adenoma 1 (2%) 1 (2%) 

Thyroid gland (50) (50) (50)

Follicular cell, adenoma 1 (2%) I (2%) 

~i!! 

General Body System 

None 

Genital System 

Epididymis 

Hemangiosarcoma, metastatic, bone marrow 

Preputial gland 

Sarcoma 

(50) 

(5o) 

(50) 

1 (2%) 

(50) 

(50) 

(50) 

1 (2%) 
314 Dietary Restriction, NTP TR 460 

TABLE Gla 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Hemat0poietic System 

Bone man:ow 

Hemangiosarcoma 

Hemangiosarcoma, metastatic, skin 

Hemangiosarcoma, metastatic, spleen 

Lymph node 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Hemangiosarcoma 

Hemangiosarcoma, metastatic, skin 

Histiocyt~c sarcoma 

Thymus 

Mediastinum, hemangioma 

Integumentary System 

Skin 

Mast cell tumor malignant 

Subcutaneous tissue, hemangiosareoma 

Musculoskeletal System 

None . 

~ 

Nervous System 

None 

Respiratory System 

Lung 

Alveolar/bwnchiolar adenoma 

Alveolar/bronchiolar adenoma, multiple 

Alveolar/bronchiolar carcinoma 

Alveolar/bronchiolar carcinoma, multiple 

Hepatocellular carcinoma, metastatic, liver 

Histiocytic sarcoma 

Nose 

Mast cell tumor malignant 

Special Senses System 

Harderian glar~ 

Adenoma 

Carcinoma 

Ad LibitumFed Control 

(50) 

1 (2%) 

(2) 

(49) 

(48) 

3 (6%) 

(50) 

I (2%) 

2 (4%) 

(42) 

(50) 

1 (2%) 

(50) 

n (22%) 

1 (2%) 

2 (4%) 

1 (2%) 

3 (6%) 

1 (2%) 

(50) 

(16) ' 

3 (19%) 

Weight-Matched 

Control 

(50) 

I (2%) 

I (2%) 

(1) 

(49) 

(43) 

(50) 

I (2%) 

(45) 

1 (2%) 

(50) 

(5o) 

9 (18%) 

1 (2%) 

1 (2%) 

2 (4%) 

(5o) 

(15) 

1 (7%) 

25 mg/kg 

(50) 

(3) 

1 (33%) 

(46) 

(47) 

1 (2%) 

(50) 

1 (2%) 

(40) 

(50) 

(50) 

5 (1o%) 

1 (2%) 

2 (4%) 

3 (6%) 

1 (2%) 

(50) 

I (2%) 

(13) 

2 (15%) 

1 (8%) 

f 
~ ~-=~-~=~ .~-~=~--~ ................. 

D~e~ry Restriction, ~ T~ ~ 31[~ 

TABLE G~a " i 

Summary o~ the IIncic~ence o~ Heoplasms h~ l~Ia~e l~,~Iice in ~he Dietary Restriction S~u~y 

of Scolpdumine Hydrobromi~e Trihydrate: Ad Libiatm ~ee~inlg and. Wdght-Ma¢che~ Contrds llhrotoeo~s (continual) 

! Ad LibitumFed Control 

Weight-Matched 

Control 25 mg/kg 

l 

t

I 

~ ,I 

I:~ [ 

~

} 

, 

~ 

[

I 

2-Year Study (continued) 

Urinary System 

Kidney 

Histiocytic sarcoma 

Artery, hepatocellular carcinoma, metastatic, 

liver 

Renal tubule, carcinoma 

Urinary bladder 

(50) ~ (2%) 

2 (4%) 

(50) 

(50) 

(50) 

(50) 

~ (2%) 

(49) 

i[ 

71 

I 

= I 

:2 i ~: I 

Systemic Lesions 

Multiple organs e 

Histiocytic sarcoma 

Leukemia lymphocytic 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

Neoplasm Summary 

Total animals with primary neoplasms a 

15-Month interim evaluation 

2-Year study 

Total primary neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with malignant neoplasms 

15-Month interim evaluation 

2-Year study 

ToTal malignant neoplasms 

i5-Month interim evaluation 

2-Year study 

Total animals with metastatic neoplasms 

2-Year study 

Total metastatic neoplasms 

2-Year study 

Total animals with uncertain neoplasmsbenign or malignant 

2-Year study 

Total uncertain neoplasms 

2-Year study 

(50) 

3 (6%) 

1 (2 %) 

1 (2%) 

3 (6%) 

4 

43 

4 

69 

4 

36 

4 

48 

17 

20 

4 

6 

I 

1 

(50) 

3 (6%) 

2 

24 

2 

34 

2 

18 

2 

22 

11 

12 

4 

5 

(50) 

I (2%} 

1 (2%) 

3 (6%) 

2 

29 

2 

37 

1 

18 

1 

20 

1 

15 

1 

17 

3 

4 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Ten animals were removed for supplemental evaluations that were not included in the dietary restriction study. 

e Number of animals with any tissue examined microscopically 

d Primary neoplasms: all neoplasms except metastatic neoplasms 

l 
316 Dietary Restriction, NTP TR 460 

TA~UE Glb / 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols a 

2-Year Restricted Feed 3-Year Restricted Feed 

Disposition Summary 

Animals initially in study 

15-MonOi intedra evaluation 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined n~ieroScopically 

15-Month Interim Evaluation • 

Endocrine System 

Thyroid gland 

Follicular cell, adenoma 

Vehicle 

Control 

60 

10 

1 

49 

60 

(10) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

Musi~uloskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

2-Year and 3-Year Protocols 

Alimentary System 

Intestine small, duodenum 

Carcinoma 

Intestine small, jejunum 

Carcinoma 

Hemangiosarcoma 

Intestine small, ileum 

Liver 

Carcinoma, metastatic, stomach, 

glandular 

Hemangiosarcoma 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

(50), 

(50) 

(49) 

(50) 

2 (4%) 

Vehicle 

25 mg/kg Control 25 mg/kg 

60 50 50 

10 

11 5 

2 11 8 

48 28 37 

60 50 50 

(lo) 

1 (10%) 

(49) (50) (5o) 

1 (2%) 

(50) (50) (49) 

1 (2%) 

2 (4%) 

(5O) (50) (49) 

(5O) (50) (50) 

1 (2%) 

1 (2%) 

7 (14%) 4 (8%) 

1 (2%) I (2%) 
• • 

Dge~ry ]Restriction, NTIP TR 460 3~7 • 

TABLE G~b 

Summary of the Incidence of Neoplasms in I~[ade Mice in the D~e~ry ~estr~cfion Study 

of Scopo]anfine Hydrobrom~de Tr~hydrate: 2-Year and 3-Year ~estr~cted Feed ]Pr'ot~coHs (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

2-Drear and 3-Year Protocols (continued) 

Alimentary System (continued) : 

Liver (continued) (50) (50) (50) (50) 

Hepatocellular adenoma 3 (6%) 7 (14%) 7 (14%) 

Hepatocellular adenoma, multiple 1 (2 %) 

Histiocytic sarcoma 2 (4%) I (2%) 

Mesentery (1) (5) (1) 

Pancreas (50) (50) (50) (50) 

Salivary glands (50) (50) (50) (50) 

Stomach, forestomach (50) (50) (50) (50) 

Squamous cell papilloma 2 (4%) 1 (2%) 1 (2%) 

Stomach, glandular (50) (50) (50) (50) 

Carcinoma 1 (2%) 1 (2%) 

Cardiovascular System 

Heart (50) (50) (50) (50) 

Carcinoma, metastatic, lung 1 (2%) 

~i I 

I 

Endocrine System 

Adrenal cortex (50) (50) (50) (50) 

Capsule, adenoma 1 (2%) 

Adrenal medulla (50) (49) 

Pheochromocytoma benign 1 (2 %) 

Islets, pancreatic (50) (49) (50) (50) 

= Adenoma 1 (2%) 1 (2%) = Pituitary gland (48) (30) (47) (47) 

Pars distalis, adenoma 3 (6%) 

Thyroid gland (50) (50) (50) (50) 

Follicular cell, adenoma 1 (2%) 1 (2%) 

General Body System 

Tissue NOB (1) 

Sarcoma l 00o%) 

Genital System 

i~: I Epididymis (50) (50) (50) (50) I

! Histiocytic sarcoma 1 (2%) 

Prostate (50) (50) (50) (50) 

Adenoma , 1 (2%) 

Seminal vesicle (50) (50) (50) (50) 

Testes (50) (50) (50) (50) 

Interstitial cell, adenoma 2 (4%) 

Histiocytic sarcoma I (2%) 
318 Dietary Restriction, NTP TR 460' 

TABLE Glb 

Summary of the Incidence of Neoplasms in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2oyear and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Hematopoietic System 

Bone marrow (50) (50) 

Hemangiosarcoma, metastatic, spleen 

Histiocytic sarcoma 

Lymph node 

Hepatocellular carcinoma, metastatic, liver 

Lymph node, mandibular (50) (49) 

Lymph node, mesenteric (50) (49) 

Carcinoma, metastatic, stomach, glandular 

Lymph node, mediastinal (1) 

Carcinoma, metastatic, lung 

Spleen (50) (50) 

Hemangioma 

Hemangiosarcoma 1 (2%) 

Histiocytic sarcoma 

Thymus (45) (45) 

Integumentary System 

Skin (50) (50) 

Subcutaneous tissue, sarcoma 1 (2%) 

Mnsculoskeletal System 

Skeletal muscle 

Nervous System 

Brain (50) (50) 

Carcinoma, metastatic, lung 

Respiratory System 

Lung (50) (50) 

Alveolar/bn)nchiolar adenoma 8 (16%) 7 (14%) 

Alveolar/bronchiolar adenoma, multiple 

Alyeolar/bronchiolar carcinoma 2 (4%) 

Alveolar/bronchiolar carcinoma, multiple 

Carcinoma, metastatic, harderian gland 

Hepatocellular carcinoma, metastatic, liver 1 (2%) 

Special Senses System 

Ear 

Histiocytie sarcoma 

Harderian gland (22) (24) 

Adenoma 2 (9%) 

Carcinoma 

3-Year Restricted Feed 

Vehicle 

Control 

(50) 

1 

1 

(4) 

1 

(48) 

(49) 

(7) 

(2%) 

(2%) 

(25%) 

1 (14%) 

(5O) 

1 (2%) 

2 (4%) 

1 (2%) 

(34) 

(48) 

(50) 

1 (2%) 

(50) 

7 (14%) 

3 (6%) 

7 (14%) 

1 (2%) 

2 (4%) 

(2) 

1 (50%) 

(24) 

2 (8%) 

1 (4%) 

25 mg/kg 

(50) 

I (2%) 

(4) 

(47) 

(46) 

1 (2%) 

(3) 

(50) 

1 (2%) 

(38) 

(50) 

(1) 

(50) 

(50) 

9 (18%) 

2 (4%) 

7 (14%) 

1 (2%) 

1 (2%) 

(23) 

4 (17%) 
D~e~ry Restriction, NTP TR 4~ . 319 

T,~E Glb 

Summary of the ]Incidence of Neopnasmsin Male Mice ~n the Dietary ~estrict~on Study 

o~ Scopo~anfine Hydrobromide Tr~hydrate: 2-Year and 3-Year ]~estr~cte~ Feed ]Protocogs (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Urinary System 

Kidney (50) (50) (50) (50) 

Carcinoma, metastatic, lung I (2%) 

Renal tubule, adenoma 1 (2%) 

Urinary bladder (50) (49) (50) (49) 

Systemic Lesions :: 

::L Multiple organs b 

Histiocytic sarcoma 

(50) (50) (50) 

3 (6%) 

(5o) 

1 (2%) 

Lymphoma malignant lymphocytic 1 (2%) 1 (2%) 5 (1o%) 4 (8%) 

Lymphoma malignant, mixed 5 (lO%) 8 (16%) 

; ; Lymphoma malignant, undifferentiated cell 1 (2%) 

Neoplasm Summa~ 

Total animals with primary neoplasms c 

• 15-Month interim evaluation 1 

2-Year and 3-year protocols 19 11 38 40 

Total primary neoplasms 

15-Month interim evaluation 1 

2-Year and 3-year protocols 23 12 59 61 

Total animals with benign neoplasms 

15-Month interim evaluation 1 

2-Year and 3-year protocols 14 9 21 24 

Total benign neoplasms 

15-Month interim evaluation 1 

2-Year and 3-year protocols 16 10 24 30 

Total animals with malignant neoplasms 

2-Year and 3-year protocols 6 2 30 29 

Total malignant neoplasms 

2-Year and 3-year protocols 7 2 35 31 

Total animals with metastatic neoplasms 

2-Year and 3-year protocols 1 6 2 

Total metastatic neoplasms 

2-Year and 3-year protocols 1 9 3 • 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

I b 

c 

Number of animals with any tissue examined microscopically 

Primary neoplasms: all neoplasms except metastatic neoplasms 
320 Dietary Restriction, NIP TR 460 

TABLE G2a . , ̧. 

Statistic~d Analysis of Primary Neoplasms in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobrotnide Trihydrate: Ad Libitura Feeding and Weight-Matched Controls Protocols 

25 mg/kg x 25 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Harderian Gland: Adenoma . 

Overall rate a 3/50 (6%) 2/50 (4%) 1/50 (2%) 2/50 (4%) 

Adjusted rate b 7.0% 4.6% 

Terminal rate c 2/40 (5 %) 1/39 (3%) 

First incidence (days) 540 405 

Life table test d P=0.511N 

Logistic regression test d P=0.441N 

Fisher exact test d P=0.500N 

Harderian Gland: Adenoma or Carcinoma 

Overall rate 3/50 (6%) 3/50 (6 %) 1/50 (2%) 3/50 (6%) 

Adjusted ral~ 7.0% 7.1% 2.4% 7.1% 

Terminal rate 2/4O (5%) 2/39 (5%) 1/41 (2%) 2/39 (5%) 

First incidence (days) 54O 405 726 (T) 405 

Life table test P=0.649 P=0.303 

Logistic regression test P=0.619N P=0.292 

Fisher exact: test P=0.661N P=0.309 

Liver: Hepatocellular Adenoma 

Overall rate 26/50 (52%) 8/50 (16%) 5/50 (10%) 8/50 (16%) 

Adjusted rate 59.0% 19.1% 11.9% 19.1% 

Terminal rate 22/40 (55%) 6/39 (15%) 4/41 (10%) 6/39 (15%) 

First incidence (days) 

Life table test 

680 587 

P<0.001N 

721 587 

P=0.259 

Logistic regression test 

Fisher exact test 

P<0.001N 

P<0.0OIN 

P=0.308 

P=0.277 

Liver: Hepatocellular Carcinoma 

Overall rate 6/50 (12%) 7/50 (14%) 5/50 (10%) 7/50 (14%) 

Adjusted rate 

Terminal rate 

14.2% 

4/40 (10%) 

17.3% 

6/39 (15%) 

11.8% 

4/41 (10%) 

17.3% 

6/39 (15%) 

First incidence (days) 

Life table test 

70O 622 

P=0.476 

532 622 

P=0.357 

Logistic regression test 

Fisher exact test 

P=0.468 

P=0.500 

P=0.413 

P=0.380 

, 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rate 30/50 (60%) 15/50 (30%) 10/50 (20%) 15/50 (30%) 

Adjusted rate 

Terminal rate 

65.2% 

24/40 (60%) 

35.3% 

12/39 (31%) 

23.2% 

8/41 (20%) 

35.3 % 

12/39 (31%) 

First incidence (days) 

Life table test • 

. 680 587 

P=0.0OTN 

532 587 

P=0.162 

Logistic regression test 

Fisher exact test 

P=0.004N 

P=0.002N 

P=0.217 

P=0.178 
-! ........ - ...... ~ : ..... 

: 

Dietary Restriction, NTP TR 460 32~ 

TABLE G2u 

S~fistlca~ Analysis of Primary Neoplasms in IV~ale 1V~ice in ~he Dietary ~estrlctlon Study 

Off Scopo~mnine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-l~,SIatched Controls Protocols (continued) 

25 mg/kg x 25 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Lung: Alveolar/bronchiolar Adenoma 

Overall rate 12/50 (24%) 6/50 (12%) 10/50 (20%) 6/50 (12%) 

Adjusted rate 28.3 % 15.4% 24.4% 15.4% 

Terminal rate 10/40 (25%) 6/39 (15%) 10/41 (24%) 6/39 (15%) 

First incidence (days) 683 726 (T) 726 (T) 726 (T) 

Life table test P=0.110N P=0.235N 

Logistic regression test P=0.120N P=0.235N 

Fisher exact test P=0.096N P=0.207N 

Lung: Alveolar/bronchiolar Carcinoma 

Overall rate 3/50 (6%) 2/50 (4%) [/50 (2 %) 2/50 (4 %) 

Adjusted rate . 6.8% 5.1%

: :l Terminal rate 1/40 (3%) 2/39 (5%) 

First incidence (days) 680 726 (T) 

Life table test P =0.519N 

Logistic regression test P=0.516N 

Fisher exact test P=0.500N 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate . 15/50 (30%) 8/50 (16%) 11/50 (22%) 8/50 (16%) 

Adjusted rate 33.8% 20.5% 26.8% 20.5% 

Terminal rate 11/40 (28 %) 8/39 (21%) 11/41 (27%) 8/39 (21%) 

First incidence (days) 680 726 (T) 726 (T) 726 (T) 

Life table test P=0.096N P=0.345N 

Logistic regression test P=0.098N P=0.345N 

Fisher exact test P =0.077N P=0.306N 

Pancreatic Islets: Adenoma 

(~verall i'ate 3/50 (6%) 1/50 (2%) 0/50 (0%) 1/50 (2%) 

Adjusted rate 7.0% 2.6% 

Terminal rate 2/40 (5 % ) 1/39 (3 %) 

First incidence (days) 647 726 (T) 

Life table test P=0.324N 

Logistic regression test P=0.313N 

Fisher exact test P= 0.309N 

All Organs: Hemangioma or Hemangiosarcoma 

Overall rate 2/50 (4%) 0/50 (0%) 3/50 (6%) 0/50 (0%) 

Adjusted rate 7.3% 0.0% " 

Terminal rate 3/41 (7%) 0/39 (0%) 

First incidence (days) 726 (T) 

Life table test P=0.130N 

Logistic regression test P=0.130N 

Fisher exact test P=0.121N 
322 Dietary Restriction, NTP TR 460 

TABLE G2a 

Statistical Analysis of Primary Neoplasms in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad L/b/turn Feeding and Weight-Matched Controls Protocols (continued) 

Ad LibitumFed Control 

All Organs: Histiocytic Sarcoma 

Overall rate 3/50 (6%) 

Adjusted rate 6.9% 

Terminal rate 2/40 (5%) 

First incid~:nce (days) 447 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Malignant Lymphoma (Lymphocytic Or Mixed) 

Overall rate 4/50 (8%). 

Adjusted rate 9.5 % 

Terminal rate 3/40 (8%) 

First incidence (days) 680 

Life table test 

Logistic regression test 

Fisher exact test 

All Organs: Benign Neoplasms 

Overall rate 36/50 (72%) 

Adjusted rate 78.2% 

Terminal rate 30/40 (75%) 

First incidence (days) 54O 

Life table test 

Logistic regression test 

Fisher exact test 

All Orgmls: Malignant Neoplasms 

Overall rate 17/50 (34%) 

Adjusted rate 36.8% 

Terminal rate 11/40 (28%) 

First incidence (days) 447 

Life table test 

Logistic regression test 

Fisher exact test 

2S mg/kg x 

AdLibitumFed Control 

1/50 (2%) 

2.5% 

0•39 (0%) 

707 

P=0.322N 

P=0.275N 

P=0.309N 

3/50 (6%) 

7.4% 

2/39 (5%) 

705 

P=0.519N 

P=0.526N 

P=0.500N 

18/50 (36%) 

42.3% 

15/39 (38 %) 

4O5 

P=0.001N 

P<0.001N 

P<0.001N 

15/50 (30%) 

34.6% 

11/39 (28%) 

4O5 

P=0.464N 

P=0.423N 

P=0.415N 

Weight-Matched 

Control 

0/50 (O%) 

3/50 (6%) 

7.3% 

3/41 (7%) 

726 (T) 

18/50 (36%) 

42.9% 

17/41 (41%) 

721 

11/50 (22%) 

26.1% 

10/41 (24%) 

532 

25 mg/kg x 

Weight-Matched 

Control 

1/50 (2%) 

3/50 (6%) 

7A% 

2/39 (5%) 

705 

P=0.641 

Pf0.661N 

P=0.661N 

18/50 (36%) 

42.3% 

15/39 (38 %) 

405 

P=0.522 

P=0.504N 

P=0.582N 

15/50 (30%) 

34.6% 

II/39 (28%) 

405 

P=0.223 

P=0.295 

P--0.247 
D[eL~ry gestr[ct~on, NTP TS 4~0 323 

=== ̧ I TABLE G2a 

St~l~st~c~d An~ys~s of Primary Neop~mrns ~n Ma~e Mice gn the D~e~ary Restriction Study 

of ~copo~zn~ne ]HIydrobrorn~de Tr~hydrz~e: Ad Libitum l~eed~ng ~nd We~ght-M~ched Controls ]Protocols (continued) 

25 mg/kg × 25 mg/kg × 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

~ ! All Organs: Benign or Malignant Neoplasms 

Overall rate 43/50 (86%) 29/50 (58%) 24/50 (48%) 29/50 (58%) 

Adjusted rate 87.8 % 62.8 % 55.8 % 62.8 % 

Terminal rate 34/40 (85%) 22/39 (56%) 22/41 (54%) 22/39 (56%) 

First incidence (days) 447 405 532 405 

Life table test P=0.018N P=0.178 • 

Logistic regression test P=0.002N P=0.304 

Fisher exact test P=0.002N P=0.212 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for liver, lung, and 

pancreatic islets; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

c Observed incidence at terminal kill 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the ad libitum-fed or weight-matched controls and the 

dosed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in the 

dosed group is indicated by N. 

e Not applicable; no neoplasms in animal group 
-- 

324 Dietary Restriction, N T P T R 460 

TA~L~g G2b 

Statistic.~d Analysis of Primary Neoplasms in Male Mice in the D i e t a r y R e s t r i c t i o n Study 

of S c o p o l a m i n e H y d r o b r o m i d e T r i h y d r a t e : 2-Year and 3-Year Restricted Feed Protocols 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

Harderian Gland: Adenoma. 

Overall rate a 2/50 (4%) 

Adjusted rate b 

Terminal rate e 

First incidence (days) 

Life table test d 

Logistic regression test d . 

Fisher exact test d 

Harderian Gland: Aden0ma or Carcinoma 

Overall rate 2150 (4%) 

Adjusted rate 

Terminal rate 

First incidence (clays) 

Life table test 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Adenoma 

Overall rate 3/50 (6%) 

Adjusted rate 6.1% 

Terminal rate • 3/49 (6%) 

First incidence (days) 722 (T) 

Life table test 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Carcinoma 

Overall rate 2/50 (4%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test, 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rate 5/50 (10%) 

Adjusted rate 10.2 % 

Terminal rate 5/49 (10%) 

First incidence (days) 722 (T) 

Life table test 

Logistic regression test 

Fisher exact test 

0/50 (0%) 

0/50 (0%) 

0/50 (O%) 

0.0% 

0/48 (0%) 

P=0.125N 

P=0.125N 

P=0.121N 

1/50 (2%) 

1/50 (2%) 

2.1% 

1/48 (2%) 

722 (T) 

P--0.109N 

P--0.109N 

P--0.102N 

-,, 

2•50 (4%) 

5.7% 

1/28 (4%) 

799 

3/50 (6%) 

8.3% 

1/28 (4%) 

799 

7/50 (14%) 

19.7% 

3/28 (11%) 

762 

7•50 (14%) 

18.3% 

2•28 (7%) 

784 

13/50 (26%) 

33.4% 

5/28 (18%) 

.762 

• 

4/50 (8%) 

10.8% 

4/37 (11% ) 

1086 (T) 

P=0.452 

P=0.348. 

P=0.339 

4/50 (8%) 

10.8%• 

4/37 (11%) 

1086 (T) 

P=0.613 

P=0.512 

P=0.500 

8/50 (16%) 

21.6% 

8/37 (22%) 

1086 (T) 

P=0.541N 

P=0.531 

P=0.500 

5/50 (10%) 

II,2% 

1/37 (3%) 

784 

P=0.320N 

P=0.413N 

P=0,380N 

13/50 (26%) 

30,9% 

9/37 (24%) 

784 

P=0.397N 

P=0.582 

P=0.590N 
-- 

: : ~ :-,~ ~ . . . . 

Dietary Restriction, N'~ TR 460 325 ~ I 

TABLE G2b 

Stafistica~ AnaHys~s of ]Primary Neoplasn~s in I~,~a~e I~¢lIice in the Dietary ]Restriction Study 

of Scopo~mnine Hydrobro~nlde Trrihydrate: 2-'Year and 3-'Year ]Restricted Feed ]?r'otoco~s (continued) 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

Lung: Alveolar/bronchiolar Adenoma 

Overall rate 8/50 (16%) 7/50 (14%) 

Adjusted ram 16.3% 14.6% 

Terminal rate 8/49 (16%) 7/48 (15%) 

First incidence (days) • 722 (T) 722 (T) 

Life table test P=0.517N 

Logistic regression test P=0.517N 

Fisher exact test P=0.500N 

Lung: Alveolar/bronchiolar Carcinoma 

Overall rate 2/50 (4%) 0/50 (0%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

=:! 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 10/50 (20%) 7/50 (14%) 

Adjusted rate 20.4% 14.6% 

Terminal rate 10/49 (20%) 7/48 (15%) 

First incidence (days) 722 (T) 722 (T) ~ ! Life table test P=0.314N 

Logistic regression test P=0.314N I 

:2;I Fisher exact test P=0.298N

I Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 0/48 (0%) 0/30 (0%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

;: I Logistic regression test i:~ [ Fisher exact test 

All Organs: Hemangioma or Hemangiosarcoma 

Overall rate 1/50 (2%) 0/50 (0%) ;;:" I Adjusted rate 

Terminal rate 

I First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

I 

3-Year Restricted Feed 

Vehicle 

Control 

' 10/50 (20%) 

28.0% 

5/28 (18%) 

620 

7/50 (14%) 

22.0%. 

5/28 (18%) 

941 

17/50 (34%) 

46.5% 

10/28 (36%) 

620 

0/47 (0%) 

0.0% 

0/27 (0%) 

4/50 (8%) 

12.7% 

3/28 (11%) 

859 

25 mg/kg 

11/50 (22%) 

28.9% 

10/37 (27 %) 

1036 

P=0.475N 

P=0.534 

P=0.500 

8/50 (16%) 

19.3% 

5/37 (14%) 

819 

P=0.561N 

P=0.515 

P=0.500 

18/50 (36%) 

43.5% 

14/37 (38%) 

819 

P=0.379N 

P=0.535 

P=0.500 

3•47 (6%) 

8.8% 

3/34 (9%) 

1086 (T) 

P=0.164 

P=0.164 

P=0.121 

1/50 (2%) 

2.7% 

1/37 (3%) 

1,086 (T) 

P=0,118N 

P=0.170N 

P=0.181N 
326 Dietary Restriction, NTP TR 460 

TABLE G2b 

Statistical Analysis of Primary Neoplasms in Male Mice in the Dietary Restriction Study. 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

All Organs: Histiocytic Sarcoma 

Overall rate 0/50 (0%) 0/50 (0%) 3/50 (6%) 1/50 (2%) 

Adjusted rate 8.8% 2.2% 

Terminal rate 0128 (0%) 0/37 (0%) 

First incidence (days) • 1,016 898

Life table test P=0.272N 

Logistic regression test P=0.310N 

Fisher exact test P=0.309N 

All Organs: Malignant Lymphoma (Lymphocytic, Mixed, or Undifferentiated Cell Type) 

Overall rate 1/50 (2 % ) 1/50 (2 % ) 11/50 (22%) 12/50 (24%) 

Adjusted rate 29.0% 28.1% 

Terminal rate 4/28 (14%) 7/37 (19%) 

First incidence (days) 784 880 

Life table test P=0.509N 

Logistic regression test P=0.497 

Fisher exact test • P=0.500

; , 

All Organs: Benign Neoplasms 

Overall rate 14/50 (28%) 9/50 (18%) 21/50 (42%) 24/50 (48 %) 

Adjusted rate 28.6% 18.8% 56.6% 61.4% 

Terminal rate 14/49 (29%) 9/48 (19%) 13/28 (46%) 22/37 (59%) 

First incidence (days) 722 (T) 722 (T) 620 892 

Life table test P=0.186N P=0.405N 

Logistic regression test P=0.186N P=0.407 

Fisher exact test P=0.171N P=0.344 

All Organ,,;: Malignant Neoplasms 

Overall rate 6/50 (12%) 2/50 (4 %) 30/50 (60%) 29/50 (58 %) 

Adjusted rate 12.0% 4.1% 66.3% 59.2% 

Terminal rate 5/49 (10%) 1/48 (2%) 13/28 (46%) 17/37 (46%) 

Firs t incidence (days) 515 5O2 784 784 

Life table test P=0.147N P=0.230N 

Logistic regression test P=0.093N P=0.501 

Fisher exact test P=0.134N P=0.500N 
327 

c 

Dietary Restriction, NT~ T~ 460 

TABLE G2b 

Stat~t~ca~ Ana~ysLs o~ ]~r~ary Neopnmms~n Mane Mice ~n the Dietary ~estr~ct~oa St~y 

o~ Scopolamine ~ydrobrom~de Tr~hydrate: 2-if ear and 3-Year ~estr~cte~ ~eed ]?r'otoco~s (con~ued) 

2-Year Restriccted Feed 3-Year ~es~cted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

All Organs: Benign or Malignant Neoplasms 

Overall rate 19/50 (38%) 11/50 (22%) 38/50 (76%) 40/50 (80%) 

Adjusted rate 38.0% 22.4% 80.6% 81.6% 

Terminal rate 18/49 (37%) 10/48 (21%) 19/28 (68%) 28•37 (76%) 

First incidence (days) 515 502 620 784 

Life table test P=0.078N P=0.257N 

Logistic regression test P=0.059N P=0.428 

Fisher exact test P = 0.063N P = 0.405 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examin~ microscopically for liver, lung, and 

pituitary gland; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence at terminal kill 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the controls and that dos~ group. The life table test 

regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in a dosed group is indicated by hi. 

e Not applicable; no neoplasms in animal group 
328 Dietary Restriction, NTP TR 460 

TABLE G3a : 

Summm'y of the Incidence of Nonneoplastic LesiOns in Male Mice in th6 Dietary Restriction Study 

of Scopolamine I-Iydrobromide Trihydrate: ,Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

,, ,?; Fed Control. Control 25 mg/kg 

Disposition Summary 

Animals initially in study 70 b 60 70 b 

15-Month inte~m evaluation " I0 lO I0 

Earlydeaths : 

Accidental deaths 1 2 

Moribund 4 2 .... 7 

Natural deaths 6 6 2 

Survivors 

Terminal sacrifice 40 41 39 

Anin~ais examined microscopically 60 60 60 . -

15-Month interim Evaluation 

Alimentary System 

Esophagus (10) (10) (10) 

Periesophageal tissue, inflammationl 

suppurative . 1 (10%) , .i 

Intestine srrmll, jejunum (10) (10) (10) t~ : 

Hyperplasia, lymphoid 1 (10~) 

Liver ........ (1o) (10) (lO) 

Basophilic focus 1 (10%) 

Clear cell focus (I0%) 

Eosinophilic focus (10%) 

Fatty change • (20%) 

Mixed cell focus (10%) 

Bile duct, hyperplasia . 00%) 

Hepatocyte, hypertrophy 

Mesentery (1) 

Fat, inflammation, chronic active 1 (100%) • ,, 

Pancreas (10) (10) (10) 

Atrophy 1 (10%) 

Atypia cellular 1 (10%) 

Stomach, forestomach (10) (10) (lO) 

Hyperplasia, focal 1 (10%) 2 (20%) 

Endocrine System 

Adrenal cortex (10) (10) (10) 

Hyperplasia 4 (40%) 2 (20%) • 2 (20%) 

Capsule, hyperplasia, adenomatous 1 (10%) 1 (10%) ,3 (30%) 

Islets, pancreatic (~0) (10) (10) 

Hyperplasia 3 (30%) 1 (10%) 

Pituitary gland (10) (9) 

Cyst .1 (10%) 

a .. Number Of animals examined microscopically at the.site and the number of animals with lesion 

b Ten_gnirnals. were remd~,eci for supplemental evaluations that werenot included in the dietary restriction study. 
? ~ - ~ ~ " ~ ~ .~- ~ e ~ _ . .__~ _ ~ . ~ ~, , ~ ~ _ ~ . ~ _ _ _ _ ~ . ~ ~ ..... 7 ~ : ~ : , ......... , , , , 

Dietary Restriction, NTP TR 460 329 

TABLE G3a 

Summary of the Incidence of Nonneopn~stic Lesions in Male Mice in the Dietzry Restriction Study 

of Scopolamine Hydrobrom~de Trihydrate: Ad Libitum Feeding and We~ght-IVI~atched Control ~FrotoeoHs (continued) 

Ad LibitumFed Control 

Weight-Matched 

Control 25 mg/kg 

~t~ i[ 

IS-Month gnterim Evaluation (continued) 

Genital System 

Preputial gland 

Duct, ectasia 

(10) 

10 (100%) 

(10) 

9 (90%) 

(10) 

9 (90%) 

Respiratory System 

Lung 

Granuloma 

Alveolar epithelium, hyperplasia 

(10) (10) 

1 (10%) 

(10) 

1 (10%) 

Special Senses System 

Eye 

Degeneration 

Harderian gland 

Inflammation, chronic active 

(4) (3) 

(1) 

1 (100%) 

(3) 

2-(67%) 

Urinary System 

Kidney 

Developmental malformation 

Nephropathy 

(lO) 

10 000%) 

(10) 

1 (10%) 

7 (70%) 

(10) 

6 (60%) 

Systems Examined With No Lesions Observed 

Cardiovascular System 

General Body System 

Hematopoietic System 

I[ntegumentary System 

I~Iusculoskeletal System 

Nervous System 

2-Year S~dy 

Alimentary System 

Esophagus 

Cyst 

Periesophageal tissue, inflammation, 

suppurative 

Intestine small, jejunum 

Hyperplasia, lymphoid 

Ulcer 

Intestine small, ileum 

Amyloid deposition 

(50) 

1 (2%) 

1 (2%) 

(50) 

(49) 

(50) 

1 (2%) 

(50) 

(50) 

(50) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 

i48) • 

1 (2%) 
330 Dietary Restriction, NTP TR 460 

TABI£ G3a 

Summ~ry of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Scol~flamine Hydrobromid~ Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

2-Year ,Study (continued) 

AHmentary System (continued) 

Liver 

Amyloid deposition 

Basophilic focus 

Clear cell focus 

Eosinophflic focus 

Fatty change. 

Hematopoieti~ cell proliferation 

" Inflammation, chronic active 

Mixed cell focus 

Necrosis 

Bile duct, cyst 

Mesentery 

.Fat,-inflammation, chronic active 

Fat, necrosis 

Pancreas 

Atrophy 

Artery, inflammation, chronic active 

~ Duct, cyst 

Salivary glands 

Atrophy 

Stomach, forestomach 

Hyperplasia, focal 

Infiltration cellular, mast cell 

Stomach, glandular 

Dysplasia 

Erosion 

Tooth 

Dysplasia 

Cardiovascular System 

Blood vessel .- 

Aorta, _ipt3arnmation, chronic active 

Heart 

Mineralization 

Artery, inflammation, chronic active 

Endocrine System. 

Adrenal cortex 

Accessory adrenal cortical nodule 

Hyperplasia 

Capsule, hyperplasia, adenomatous 

Adrenal n~edulla ..... 

Inflammation, chronic active 

Islets, pancreatic. 

Hyperplasia 

(50) 

3 (6%) 

12 (24%) 

21 (42%) 

I (2%) 

2 (4%) 

1 (2%) 

5 (10%) 

1 (2%) 

(4) 

2 (50%) 

(50) 

2 (4%) 

(50) 

(50) 

3 (6%) 

(50) 

(14) 

13 (93%) 

(50) 

(50) 

2 (4%) 

(50) 

1 (2%) 

23 (46%) 

7 (14%) 

(49) 

1 (2%) 

(50) 

29 (58%) 

(50) 

2 (4%) 

I (2%) 

2 (4%) 

1 (2%) 

I (2%) 

(I) 

l (I00%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

16 (32%) 

1 (2%) 

(50) 

I (2%) 

5 (10%) 

(50) 

(50) 

(50) 

1 (2%) 

10~(20%) 

6 (12%) 

(50) 

(50) 

2 (4%) 

(50) 

1 (2%) 

2 (4%) 

2 (4%) 

1 (2%) 

1 (2%) 

3 (6%) 

(2) 

1 (50%) 

(50) 

i (2%) 

2 (4%) 

(50) 

(50) 

6 (12%) 

(50) 

(2) 

I (50%) 

(50) 

1 (2%) 

(50) 

I (2%) 

(50) 

1 (2%) 

25 (50%) 

3 (6%) 

(50) 

(50) 

2 (4%) 
Dietary Restriction, NTP TR 460 33~. 

TABLE G3a 

Sunamm'y of the ]Incidence of Nonneopn~stic Lesions in Male Mice in the Diet~'y Restriction Study 

of Scopoimnine Hydrobronfide Trrihydrate: Ad Libitum ~eeding and Weight-Matched Controls ~Pr'otecolls (continued) 

2- ~rear Study (continued) 

Endocrine System (continued) 

Pituitary gland 

Pars distalis, hyperplasia 

Pars intermedia, hyperplasia 

Thyroid gland 

Follicular cell, hyperplasia 

General Body System 

None 

~ I GenRal System 

Epididymis 

Granuloma sperm 

Inflammation, chronic active 

Preputial gland 

Inflammation, chronic active 

Duct, ectasia 

Prostate 

Inflammation, chronic active 

Seminal vesicle 

Inflammation, chronic active 

Testes 

Atrophy 

Hypoplasia 

Interstitial cell, hyperplasia 

Hematopoietic System 

Bone marrow 

Thrombosis 

Erythroid cell, hyperplasia 

Mycloid cell, hypcrplasia 

i!E [ 

Lymph node 

Lumbar, hypcrplasia, lymphoid 

Mcdiastinal, hypcrplasia, lymphoid 

Lymph node, mandibular 

Hyperplasia, lymphoid 

Lymph node, mcscnteric 

Hcmatopoietic cell proliferation 

Hypcrplasia, lymphoid 

Hypcrplasia, plasma cell 

Ad LibitumFed Control 

(48) 

1 (2%) 

(50) 

13 (26%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

47 (94%) 

(49) 

(50) 

1 (2%) 

(50) 

2 (4%) 

(50) 

1 (2%) 

(2) 

1 (50%) 

(49) 

(48) 

2 (4%) 

1 (2%) 

Weight-Matched 

Control 

(48) 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

47 (94%) 

(50) 

(50) 

(50) 

1 (2%) 

(50) 

5 (10%) 

1 (2%) 

(1) 

(49) 

(43) 

25 mg/kg 

(44) 

1 (2%) 

(50) 

5 (10%) 

(50) 

1 (2%) 

(50) 

3 (6%) 

45 (90%) 

(50) 

2 (4%) 

(50) 

(50) 

1 (2%) 

(5o) 

1 (2%) 

2 (4%) 

5 (lO%) 

(3) 

1 (33%) 

(46) 

1 (2%) 

(47) 

I (2%) 

i (2%) 
332 Dietary Restriction, NTP TR 460 

TABLE G3a 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued~ 

2-Year Study (continued) 

Hematopoietie System (continued) 

Spleen 

Amyloid deposition 

Angiectasis 

Depletion lymphoid 

Hematopoietic cell proliferation 

Hyp~rplasia, lymphoid 

Thymus 

Atrophy 

Epithelial cell, hyperplasia 

Integumentary System 

Skin 

Inflammation, chronic active 

Ulcer 

Subcutaneous tissue, lymphangiectasis 

Musculoskeletal System 

None . 

Nervous System 

Brain 

Neuron, necrosis 

Respiratory System 

Lung 

Embolus 

Inflammation, chronic active 

Inflhmmation, suppurati~,e 

Alveolar epithelium, hyperplasia 

Nose 

Inflammation, suppurative 

Special Senses System 

E~,e 

Cornea, inflammation, chronic active 

Harderian gland 

Hyperplasia 

Ad LibitumFed Control 

(50) 

1 (2%) 

8 (16%) 

I (2%) 

(42) 

6 (14%) 

I 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 

8 (16%) 

(50) 

(16) 

1 (6%) 

Weight-Matched 

Control 

(50) 

1 (2%) 

5 (10%) 

7 (14%) 

(45) 

5 (11%) 

I (2%) 

(50) 

<50) 

1 (2%) 

(50) 

5 (10%) 

(50) 

(15) 

, 

2S mg/kg 

(5O) 

I (2%) 

3 (6%) 

12 (24%) 

(40) 

7 (18%) 

(50) 

l (2%) 

I (2%) 

(50) 

(50) 

I (2%) 

1 (2%) 

1 (2%) 

(50) 

2 (4%) 

(2) 

1 (50%) 

(i3) 
~L~ 

D~ef~ry Res~:icfion, I~ITP TR ,~60 333

T~LE G3a 

S~n~ary of the Incidence of Nonneop~astic Lesions in I~Ia~e Mice in the Dietary Restriction Stuc~y 

of Scopolan~ine Hy~robron~i~e Tr~hy~rate: Ad Libitum Feeding anc~ We~ght-iV~atche~ Controls Protocols, continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

2.Year Study (continued) 

Urinary System 

Kidney (50) (50) (50) 

Cyst 6 (12%) 3 (6%) 3 (6%) 

Hydronephrosis 1 (2%) 

Inflammation, chronic active 2 (4%) 2 (4%) 

Necrosis 1 (2%) 

Nephropathy 48 (96%) 42 (84%) 37 (74%) 

Glomerulus, amyloid deposition 1 (2 %) 

Urinary bladder (50) (50) (49) 

Inflammation, chronic active 2 (4%) 3 (6%) 
334 Dietary Restriction, NTP TR 460 

TABLE G3b 

Summary of the Incidence of Nonneoplastic LesionsJn Mal e Mice in the Dietary Restriction Study , 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols a 

Disposition Summary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Moribund. 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

Alimentary System 

Stomach, forestomach 

Hyperplasia, focal 

Stomach, glandular 

Dysplasia 

Cardiovascular System 

Heart 

Inflammation, chronic active 

Endocrine System 

Adrenal codex 

Accessory adrenal cortical nodule 

Pituitary gland 

Cyst 

Hematopoietic System 

Bone marrow 

Hyperplasia, mast cell 

Special Senses System 

Harderian gland 

Inflammation, chronic active 

Urinary System 

Kidney 

Hyperplasia, mast cell 

Nephropathy 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

60 60 

10 10 

1 2 

49 48 

60 60 

(10) (10) 

6 (60%) 6 (60%) 

(10) (10) 

1 (10%) 

(10) (10) 

1 (10%) 

(10) .(10) 

2 (20%) 

(10) (8) 

1 (13%). 

(10) (10) 

1 (10%) 

(4) (2) 

2 (50%) 2 (100%) 

(10) (10) 

1 (10%) 

9 (9o%) 5 (5o%) 

3-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

5O " ̧ . 50 

11 5 

11 8 

28 37 

50 50 

a Number of animals examined microscopically at the site and the number of animals with lesion 
Di~zy ~esCr~ction, ~ TR ~0 335 

TA~L~ G3b 

S~nmry of the ]Incidence o~ Honneo~l~s~ic Lesions ~ I~e IV~ce in ~he Die~n-y Res~r~cfio~ S~y 

o~ Sco~o~i~e Hy~ro~ro~i~e Tri~y~r~e: ~-Ye~r ~n~ ~-Ye~r Re~r~c~e~ ~ee~ ProCtors (con~uod) 

2-Year ]Restricted Feed 3-Year ]Restricted Feed 

Vehicle Vehicle 

Control 25 m~/~ Control 25 mg/kg 

15-Month #nterbn Evalu~on (continued) 

Systems Examined With No Lesions Observed 

General Body System 

Genital System 

integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

2-Year and 3-Year Proeocols 

Alimentary System 

Esophagus (50) 

Periesophageal tissue, inflammation, 

suppurative 1 (2%) 

Gallbladder (50) 

Inflammation, acute 

Intestine small, jejunum (50) 

Hyperplasia, lymphoid 1 (2%) 

Inflammation, chronic active 

Liver (50) 

Basophilie focus 

Clear cell focus 1 (2%) 

Eosinophilic focus 1 (2%) 

Hematopoietic cell proliferation 

Hepatodiaphragmatic nodule 

Hyperplasia, lymphoid 

Mixed cell focus 

Necrosis 1 (2%) 

Bile duct, cyst 

Bile duct, hyperplasia, cystic 1 (2%) 

Mesentery (1) 

Fibrosis ,1 (100%) 

Artery, inflammation, chronic, active 

Fat, inflammation, chronic active 

Fat, necrosis 

Pancreas (50) 

Inflammation, chronic active 

• Acinus, atrophy 

Aeinus, hyperplasia, focal 

Artery, inflammation, chronic active 

Duet, ectasia 

Stomach, forestomach (50) 

Cyst 

Hyperplasia, focal 38 (76%) 

Hyperplasia, mast cell 

(50) 

(49) 

(50) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

(50) 

32 (64%) 

(50) 

(50) 

(50) 

I (2%) 

, I (2%) 

(50) 

! (2%) 

! (2%) 

1 (2%) 

I (2%) 

2 (4%) 

2 (4%) 

I (2%) 

(.5) 

3 (60%) 

2 (40%) 

(50) 

I (2%) 

2 (4%) 

2 (4%) 

1 (2%) 

(5O) 

25 (50%) 

(50) 

(49) 

1 (2%) 

(49) 

5 (lO%) 

(5o) 

1 (2%) 

l (2%) 

2 (4%) 

(1) 

1 000%) 

(50) ' 

1 (2%) 

(50) 

2 (4%) 

33 (66%) 

3 (6%) 
336 Dietary Restriction, NTP TR ~ 

, "'r 

TABLE G3b • ~ : • 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study • 

of Scopo]lamine Hydrobromide Trihydrate: 2-Year and 3=Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Alimentary System (continued) 

Stomach, glandular (50) (50) 

Dysplasia 1 (2%) 1 (2%) 

Hyperplasia 

Tongue ( 1 ) 

Mineralization 1 (100%) 

Tooth ' (2) 

Dysplasia 2 (100%) 

Inflammation, chronic active 

Cardiovascular System 

Heart . (50) (50)

Degeneration 

Mineralization 

Atrium, thrombosis 

Vale, e, inflammation; chronic active 

Endocrine System 

Adrenal cortex (50) (50) 

Accessory adrenal cortical nodule 2 (4%) 

Hematopoietic cell proliferation 

Hemorrhage 

Hyperplasia 14 (28%) 6 (12%) 

Capsule, hyperplasia 

Capsule, hyperplasia, adenomatous 6 (12%) 

Adrenal medulla (50) (49) 

Hyperplasia 

Islets, pancreatic " (50) (49) 

Hyperplasia 

Parathyroid gland 

1 (2%) 

(42) (44) 

Cyst 

Hyperplasia, focal 

Pituitary gland (48) (30) 

Pars distalis, cyst 

Pars distalis, hyperplasia 

Thyroid gland (50) (50) 

Inflammation 

Inflammation, chronic active 

Follicular cell, hyperplasia 1 (2%) 

General Body System 

None 

3-Year Restricted Feed 

'Vehicle 

control 25 mg/kg 

(50) (50) • 

1 (2%) 3 (6~) 

1 (2%) 

(I) 

(3) (7) 

3 (100%) 6 (86%) 

1 (14%) 

(5o) (50) 

1 (2%) 

1 (2%) 1 (2%) 

1 (2%) 

1 (2%) 

(50) (50) 

1 (2%) 1 (2%) 

1 (2%) 

1 (2%) 

6 (12%) 12 (24%) 

3 (6%) 1 (2%) 

. (50) (50) 

3 (6%) 1 (2%) 

(50) (50) 

3 (6%) 

(50) (47) 

I. (2%) 

1 (2%) 

(47) (47) 

3 (6%) 1 (2%) 

2 (4%) 3 (6%) 

(50) (50) 

1 (2%) • . 

1 (2%) 

6 (12%) 2 (4%) ;. 
337 

7:: 7 ~ ~=~ : ~?~:~7~ .................... ................... ~ ~ ~ ...... ~- ............... ~ 

Dietary Restriction, NT~ TR 460 

TABLE G3b 

Summary o[ the ]Incidence of Nonneoplasfic Lesions ~n Male Mice ~n the D[etm'y ~estr~ct[on Study 

o~ Scopolamine Hydrobrornide Tr!hydrate: 2-Year and 3-Year Restricted Feed ~'r'otoco~s (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mglkg 

2.Year and 3-Year Protocols (continued) 

Genital System 

Epididymis (50) (50) (50) (50) 

Granuloma sperm 7 (14%) 6 (12%) 

Inflammation, chronic active 1 (2%) 

Spermatocele 1 (2%) 

Preputial gland 

Degeneration 

(50) (49) (50) (50) 

1 (2%) 

Inflammation, chronic active 2 (4%) 1 (2%) 7 (14%) 6 (12%) 

Duct, ectasia 24 (48%) 10 (20%) 24 (48%) 15 (30%) 

Testes (50) (50) (50) (50) 

Atrophy- 1 (2%) 9 (18%) 7 (14%) 

Hypoplasia 1 (2%) 

Interstitial cell, hyperplasia 1 (2%) 

Hematopoietie System 

Bone marrow 

Myelofibrosis 

(50) (50) (50) 

2 (4%) 

(50) 

Erythroid cell, hyperplasia 

Myeloid cell, hyperplasia 6 (12%) 

3 (6%) 

1 (2%) 

Lymph node, mandibular 

Hyperplasia, lymphoid 

(50) (49) (48) (47) 

1 (2%) 

Lymph node, mesenteric 

Angiectasis 

(50) (49) (49) 

5 (10%) 

(46) 

5(11%) 

I 

~ I 

i ! 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Spleen 

Depletion lymphoid 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

(50) 

2 (4%) 

(50) 

1 (2%) 

1 (2%) 

(5O) 

2 

15 

1 

(4%) 

(30%) 

(2%) 

2 (4%) 

1 (2%) 

(50) 

I (2%) 

11 (22%) 

1 (2%) 

Hyperplasia, plasma cell 1 (2%) 

Thrombosis 1 (2%) 

Thymus (45) (45) (34) (38) 

ii~ i Atrophy 1 (2%) 13 (38%) 9 (24%) 

I 

Integumentary System 

Skin (50) (50) (48) (5o) 

Subcutaneous tissue, inflammation, 

chronic active 2 (4%) 

Musculoskeletal System 

None 
338 Dietary Restriction, NTP TR 460 

TABLE G3b 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Nervous System 

Brain 

Infdtration cellular~ lymphocyte 

Respiratory System 

Lung 

Bronchiectasis, focal 

Inflammation 

Alveolar epithelium, hyperplasia . 

Bronchiole, hyperplasia 

Nose 

Respiratoz3~ epithelium, inflammation, 

chronic active 

Special Senses System 

Eye 

Lens, cataract 

Harderian gland 

Hyperplasia 

Urinary System 

Kidney 

Cyst 

Necrosis, focal 

Nephropathy 

Renal tubule, hyperplasia 

(50) (50) (50) (50) 

1 (2%) 

/ 

(50) (50) (50) (50) 

1 (2%) 

1 (2%) 

5 (10%) I (2%) 4 (8%) 7 (14%) 

1 (2%) I (2%) 

(50) (50) (50) (50) 

1 (2%) 

O)

1 (I00%) 

(3) 

2 (67%) 

(22) (24) (24) (23) 

1 (5%) . 

(50) (50) (50)

3 (6%) 

(50) 

1 (2%) 

1 .(2%) 

47 (94%) 

1 (2%) 

45 (90%) 4O (80%) 4O (80%) 

l (2%) 
." ' • 339 

APPEHI}]IX H 

SUMMAI~IY OF LESI©HS IN FEMALE MICE 

~IN THE I~ETAXY XESTI~ICT~©N STILr~Y 

©IF SC©P©LAI~dI~HE HYI~©B~©M~I~E TI~HYI~I~ATE 

TABLE Hla Summary of the Incidence of Neoplasms in Female Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 340 

TABLE Hlb Summary of the Incidence of Neoplasms in Female Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

2-Year and 3-Year Restricted Feed Protocols .................... : ......... 345 

TABLE I-I2a Statistical Analysis of Primary Neoplasms in Female Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 350 

TABLE ]~b Statistical Analysis of Primary Neoplasms in Female Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

2-Year and 3-Year Restricted Feed Protocols .............................. 354 

TABLE H3a Summary of the Incidence of Nonneoplastic Lesions in Female Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols .................... 359 

TABLE H3b Summary of the Incidence of Nonneoplastic Lesions in Female Mice 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

2-Year and 3-Year Restricted Feed Protocols .............................. 364 
340 Dietary Restriction, NTP TR 460 

TABLE Hla 

Summary of the Incidence of Neoplasms in Female Mice in the Dietary Restriction Study 

of Scol~alamine Hydrobromide Trihydrate: Ad Libitum Feeding and. Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

Disposition Summary 

Animals irfitially in study 70 b 60 70 b 

15-Month inte~m evaluation I0 I0 10 

Early deaths 

Accidental deaths 2 1 

Moribund 9 10 7 

Natural deaths 7 3 6 

Survivors 

Terminal sacrifice 33 36 38 

Animals examined microscopically 61 60 61 

15-Month Interim Evaluation 

Alimentary System 

Liver (10) (10) (10) 

Hepatocellular adenoma 1 (10%) 

Endocrine System 

Thyroid ghnd 0o) (10) 0o) 

Follicular cell, adenoma ~ (~o%) 

Genital System 

Ovary (10) (lO) (~o) 

Cystadenoma 1 (10%) 

Systetns Examined With No Neoplasms Observed 

Cardiovascular System 

General Body System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 
341 

TABLE Hit 

Summary of the Incidence of Neoplasms in Female 1~,SIice ~n the Dietary IRestriction Study 

of Scopolamine Hydrobrom[de Tr~hydrate: Ad Libitum Feeding and We~ght-lV~atched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

2-Year Study 

Alimentary System 

Gallbladder (51 ) (50) (51 ) 

Intestine small, jejunum (51) (49) (50) 

Carcinoma 1 (2%) 

Liver (51) (50) (51) 

Hemangiosarcoma, multiple 1 (2%) 

Hepatocellular carcinoma 7 (14%) 2 (4%) 4 (8%) 

Hepatocellular carcinoma, multiple 1 (2 %) 

Hepatocellular adenoma 10 (20%) 7 (14%) 4 (8%) 

Hepatocellular adenoma, multiple 5 (10%) 2 (4%) 

Hepatocholangiocarcinoma 1 (2%) 

Histiocytic sarcoma I (2%) 3 (6%) 1 (2%) 

Bile duct, carcinoma 1 (2%) 

Mesentery (7) (5) (5) 

Carcinoma, metastatic, liver 1 (14%) 

Histiocytic sarcoma 1 (20%) 1 (20%) 

Lipoma 1 (20%) 

Pancreas (51 ) (50) (51 ) 

Carcinoma, metastatic, liver 1 (2%) 

Histiocytic sarcoma 1 (2%) 

Salivary glands (51) (50) (51) 

Stomach, forestomach (51) (50) (51) 

Squamous cell papilloma 1 (2%) 2 (4%) 

Stomach, glandular (51) (50) (51) 

Tongue (1) 

Squamous Cell carcinoma 1 (100%) 

Cardiovascular System 

Heart (51) (50) (51) 

Carcinoma, metastatic, liver 1 (2%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (2%) 

Endocrine System 

Adrenal cortex (51) (50) (51) 

:;~; I Adrenal medulla 

Pheochromocytoma malignant 

(51) 

1 (2%) 

(49) (50) 

Pituitary gland (50) (48) (46) 

;;I Pars distalis, adenoma 

Pars interrnedia, adenoma 

4 (8%) 1 (2%) 3 (7%) 

1 (2%) 

Thyroid gland (51) (50) (51) 

Follicular cell, adenoma 3 (6%) 1 (2%) 2 (4%) 

General Body System 

None 
342 Dietary Restriction, NTP TR 460 

TABLE llla 

Summat T of the Incidence of Neoplasms in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and weight-Matched Controls Protocols (continued) 

2-Year Study (continued) 

Genital System 

Ovary 

Cystadenoma 

Hemangiosarcoma 

Histiocyfic sarcoma 

Luteoma 

Uterus 

Adenoma 

Hemangiosarcoma 

Hemafigiosarcoma, metastatic, ovary 

Histiocytic Sarcoma 

Neoplasm NOS 

Polyp stromal 

Sarcoma stromal 

Hematopoietic System 

Bone marrow 

Hemangiosarcoma 

Histiocytic sarcoma . 

Lymph node 

Bronchial, carcinoma, metastatic, liver 

Bronchial, hepatocholangiocarcinoma, 

metastatic, liver 

Bronchial, histiocytic sarcoma 

Mediastinal, carcinoma, metastatic, liver 

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, histioeytic sarcoma 

Lymph node, mandibular 

Histiocytic sarcoma 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Hemangiosarcoma 

Histiocytic sarcoma 

Thymus 

Hepatocholangiocarcin0ma, metastatic, liver 

Histiocytic sarcoma 

Mast cell tumor malignant 

Int,egumentary System 

Skin 

Squamous cell carcinoma 

Subcutaneous tissue, fibrosarc~ma 

Ad LibitumFed Control 

(51) 

3 (6%) 

1 (2%) 

(51) 

i (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

(51) 

1 (2%) 

(6) 

1 (17%) 

1 (17%) 

I (17%) 

1 (17%) 

1 (17%) 

(50) 

(50) 

(51) 

1 (2%) 

1 (2%) 

(48) 

1 (2%) 

I (2%) 

(51) 

i (2%) 

Weight-Matched 

Control 

(49) 

2 (4%) 

(50) 

1 (2%) 

2 (4%) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

(3) 

1 (33%) 

1 (33%) 

(48) 

1 (2%) 

(48) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 

(43) " 

1 (2%) 

(50) 

:~ mg/kg 

(51) 

1 (2%) 

1 (2%) 

(51) 

(51) 

1 (2%) 

(3) 

1 (33%) 

(51) 

(48) 

(51) 

1 (2%) 

(45) 

I (2%) 

(51) 

I (2%) 
Dietary IRestr~etion, NT]? TI~ ~0 3~ 

T~L~ ~a 

Sun~ary o~ the l~acidence o~ Neop~sn~s ~n ]~en~aHe I~,]Iice ~n the Dietary ~estr~ct~on Study 

o~' Scopogan~ne ~ydrobro~n~de Tr~hydrate: Ad L~b/t~m ~eed~ng and We~ght-I~,~atched Controls ]Prot~co]Is (¢on~ued) 

Ad LibitumFed Control 

Weight-Matche~l 

Control 25 mg/kg 

:! 2-Drear S~dy (continued) 

Museuloskeletal System 

Bone 

Hepatocholangiocarcinoma, metastatic, liver 

Skeletal muscle 

Hepatocholangiocarcinoma, metastatic, liver 

Rhabdomyosarcoma 

(51 ) 

1 (2%) 

(1) 

1 (100%) 

(50) (51) 

Nervous System 

Brain (50) (51) 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, harderian gland 

Carcinoma, metastatic, liver 

Hemangiosarcoma, metastatic, ovary 

Hepatocellular carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, liver 

Histiocytic sarcoma 

Mediastinum, hemangiosarcoma 

Nose 

Histiocyticsarcoma 

(51) 

3 (6%) 

1 (2 %) 

1 (2%) 

I (2%) 

1 (2%) 

1 (2%) 

I (2%) 

I (2%) 

(51) 

(50) 

2 (4%) 

I (2%) 

2 (4%) 

~ (2%) 

• (50) 

1 (2%) 

(51) 

2 (4%) 

I (2%) 

I (2%) 

(51) 

Special Senses System 

Harderian gland 

Adenoma 

Carcinoma 

(17) 

1 (6%) 

2 (12%) 

(I0) 

3 (3O%) 

(13) 

I (8%) 

Urinary System 

Kidney 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, liver 

Histiocytic sarcoma 

Urinary bladder 

(51 ) 

1 (2%) 

I (2%) 

(50) 

(50) 

1 (2%) 

(50) 

(51 ) 

(5 I) 

Systemic Lesions 

Multiple organs c 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

(51) 

I (2%) 

2 (4%) 

7 (14%) 

(50) 

4 (8%) 

I (2%) 

5 (10%) 

(51) 

1 (2%) 

4 (8%) 

3 (6%) 
344 Dietary Restriction, NTP TR 460 

TABLE Ilia 

Summary of the Incidence of Neoplasms in Female Mice in the Dietary Restriction .Study 

of_Scopolamine Hydrobromide Trihydrate: AdLibitum Feeding and Weight-Matched Controls Protocols ~continued) 

• Ad Libitum- Weight-Matched 

~'~ Fed Control Control 25 mg/kg 

Neoplasm Summary .._. 

Total animals with primary ~oplasms d 

15-Month interim evaluation 1 1 

2-Year study 40 28 29 

Total primary neoplasms 

15-Month interim evaluation 2 1 

2-Year study 62 35 35 

Total animals with benign neoplasms 

15-Month interim evaluation 1 1 

2-Year study 24 14 18 

Total benign neoplasms 

15-Month interim evaluation 1 

2-Year study . 31 17 19 

Total animals with malignant neoplasms 

2-Year study ~ 26 16 14 

Total malignant neoplasms 

2-Year study 31 17 16 

' Total animals with metastatic neoplasms 

2-Year study 5 1 

Total metastatic neoplasms 

• 2-Year study 19 1 

Total animals with uncertain neoplasmsbenign or malignant 

2:Year study 

Total uncertain neoplasms 

2-Year study 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Nine animals were removed for supplemental evaluations that were not included in the dietary restriction study. 

c Number of animals with any tissue examined microscopically 

d Primary neoplasms: all neoplasms except metastatic neoplasms 

._ 
~ 

Dietary Restriction, ~ T~ ~6~ 34S 

TA~ Hlb 

Sun~n~ry of the Incidence o~ Heop~snm in ]Fem~de lV~ice in the Dietary ~est~ction Study 

o~ ScoIml~nine Hydrobromide Trihydrate: 2-Year ~nd 3-Year Restricted ]Feed ]?r'ot~co~s a 

2,.-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 ms/ks 

Disposition Summary 

Animals initially in study 60 60 

15-Month interim evaluation 10 10 

, Early deaths 

Accidental deaths 1 

Moribund 2 2 

Natural deaths 1 3 

Survivors 

Terminal sacrifice 47 44 

Animals examined microscopically 

Systems Examined At 15 Months With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

Endocrine System 

General Body System 

Genital System 

Hematopoietic System 

Integumentary System 

lV~usculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

2-3rear and 3-3rear Protocols 

Alimentary System 

Gallbladder (50) (50) 

Intestine large, cecum (50) (50) 

Intestine small, jejunum (50) (50) 

Histiocytie sarcoma 

Sarcoma, metastatic, skin 

Liver (50) (50) 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 3 (6%) 3 (6%) 

Hepatocellular adenoma, multiple 

Histiocytic sarcoma 1 (2 %) 3 (6%) 

Osteosarcoma, metastatic, bone 1 (2%) 

Mesentery (2) 

Hemangiosarcoma 

Histiocytic sarcoma 1 (50%) 

Sarcoma, metastatic, skin 

Pancreas (50) (50) 

Sarcoma, metastatic, skin 

(50) 

(49) 

(50) 

2 

(50) 

3 

1 

11 

1 

4 

(~ 

1 

2 

2 

(49) 

50 

18 

12 

20 

50 

(4%) 

(6%) 

(2%) 

(22%) 

(2%) 

(8%) 

(17%) 

(33%) 

(33%) 

(49) 

(48) 

(49) 

1 

(50) 

4 

7 

1 

5 

(6) 

2 

2 

(50) 

1 

50 

1 

16 

14 

19 

50 

(2%) 

(8%) 

(14%) 

(2%) 

(10%) 

(33%) 

(33%) 

(2%) 
346 Dietary Restriction, NTP TR 460 

TAnt~ Hlb 

Summary of the Incidence of Neoplasms in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (tongued) ~ ~ , • 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Alimentary Systdm (continued) 

Salivary gland~ (50) (50) (49) (50)

Stomach, forestomach (50) (50) (50) (50)

Squamous cell papilloma l (2%)

Stomach, glandular (50) (50) (50) (50)

Tooth (2) (2)

Nistiocytic sarcoma 1 (50%) 

Cardiovascular System 

Heart 

Pericardium, osteosareoma, metastatic, bone 

(50) (50) 

1 (2%) 

(50) (50) 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Carcinoma 

Pituitary gland 

Histiocytie sarcoma 

Pars distalis, adenoma 

(49) 

(49) 

1 : 

(50) 

(46) 

,. 

(2%) 

(50) 

(50) 

I (2%) 

(5o) 

(46) 

1 (2%) 

._ 

(50) 

(49) 

l (2%) 

(49) 

1 (2%) 

2 (4%) 

(42) 

3 (7%) 

(50) 

(50) 

1 

(50) 

2 

(49) 

! 

2 

(2%) 

(4%) 

(2%) 

(4%) 

Pars intermedia, adenoma 

Thyroid gland . 

1 

(50) 

(2%) 

(49) 

4 (10%) 

(50) 

2 

(50) 

(4%) 

Bilateral, follicfiiar cell, adenoma 1 (2%) 

Follicular cell, adenoma 1 (2%) 1 (2%) 

Follicular cell, carcinoma . ." (2%) 

General Body System 

None 

Genital System 

Ovary 

Adenoma 

~ (50) (50) (49) (49) 

1 (2%) 

Adenoma, tubular .. 1 (2%) 

Cystadenoma 1 (2%) 2 (4%) 

Granulosa cell tumor be~gn 1 (2%) 

Histiocytic sarcoma 1 (2%) 3 (6%) 2 (4%) 

Sarcoma, metastatic, skin 1 (2%) 

Uterus (50) (50) (49) (49) 

Histiocytic sarcoma 2 (4%) 1 (2%) 4 (S%) 3 (6%) 

Polyp stromal 2 (4%) 1 (2%) 2 (4%) 
....... - .............................. .~ ...... ~ ....... = ~--. ~ ~=.~2 ................ ~ ~ 

Die~ry Restriction, NTI? TR ~0 

TABLE Hlb 

Summary o~ the ~ncidence o~ Neop~asnm ~n Fe~na~e Mice ~n the Dietary Restriction Study 

o~ Scopo~m~ne Hydrobrom~de ']rr~hydrate: 2-Ye~" ~d 3-YeAr ~estr~cted Feed l?rot~o~s (eon~ued) 

2-Year Restricted Feed 3-Ye~" Restricted Feed 

Vehicle Vehicle 

Control 2s mg/kg Control 2~ mf,/k~ 

2-Year and 3-Year Protocols (continuod) 

Hematopoietic System 

Blood (i) 

Bone marrow (50) (50) (50) (5o) 

Hemangiosarcoma 2 (4%) I (2%) 

Hemangiosarcoma, multiple 1 (2%) 

Histiocytie sarcoma 1 (2%) 1 (2%) i (2%) 1 (2%) 

Lymph node (13) (8) 

Hepatocellular carcinoma, metastatic, liver 1 (8%) 

Histiocytic sarcoma 2 (15%) 

Axillary, sarcoma, metastatic, skin I (8%) 

Pancreatic, sarcoma, metastatic, skin I (8%) 

Lymph node, bronchial 

Lymph node, mandibular (50) 

(1) 

(48) (45) 

(1) 

(47) 

Histiocytic sarcoma 1 (2%) 1 (2%) 1 (2%) 

Mast cell tumor NOS 1 (2%) 

Lymph node, mesenteri¢ 

Histiocytic sarcoma 

Lymph node, mediastinal 

(44) 

1 (2%) 

(2) 

(50) 

2 (4%) 

(4) 

(43) 

2 

(i2) 

(5%) 

(48) 

3 (6%) 

(11) 

Histiocytic sarcoma 1 (50%) 2 (50%) 1 (8%) 2 (18%) 

Sarcoma, metastatic, skin 1 (8%) 

Spleen (50) (50) (50) (50) 

Hemangiosarcoma 

Histiocytic sarcoma 

Thymus (45) 

2 (4%) 

(49) 

1 

1 

(37) 

(2%) 

(2%) 

3 (6%) 

(42) 

Alveolar/bronchiolar carcinoma, metastatic, 

lung 1 (2%) 

Histiocytic sarcoma (5%) 

Osteosarcoma, metastatic, bone 1 (2%) 

~ntegumentary System 

Mammary gland (50) (50) (49) (49) 

Carcinoma 1 (2%) 

Skin (50) (50) (5o) (50) 

Basal cell adenoma 1 (2%) 

Squamous cell carcinoma I (2%) 

Subcutaneous tissue, fibrosarcoma 1 (2%) 

Subcutaneous tissue, sarcoma 4 (8%) 5 (1o%) 

Musculoskeletal System 

Bone (50) (50) (5o) (5O) 

Ostcosarcoma I (2%) 

Vertebra, ostcosarcoma i (2%) 

347 

I 

I 
348 Dietary Restriction, NTP TR 460 

TABLE Hlb 

Summary of the Incidence of Neoplasms in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

/~ 

2-Year t~nd 3-Year Protocols (continued) 

Nervous System 

Brain (50) (50) 

Ast~ocytoma malignant 

Histiocytie sarcoma 

Respiratory System 

Lung • (50) (50) 

Alveolar/bronchiolar adenoma 3 (6 % ) 2 (4%) 

Alveolar/bronchiolar carcinoma 1 (2 %) 

Alveolar/bronchiolar carcinoma, multiple 2 (4%) 

Hepatocellular carcinoma, metastatic, liver 

Histiocyfic sarcoma 1 (2 %) 1 (2%) 

Osteosarcoma, metastatic, bone 1 (2%) 

Mediastinum, aiveolar/bronchiolar 

carcinoma, metastatic, lung 1 (2%) 

Nose (50) (49) 

Special Senses System 

Harderian gland (24) (23) 

Adenoma 3 (13%) 

Bilateral, adenoma 

Urinary System 

Kidney (50) (50) 

Histiocytic sarcoma 1 (2%) 

Osteosarcoma, metastatic, bone 

Urinary bladder (50) (50) 

Histiocytic sarcoma 

Systemic Lesions 

Multiple organs b (50) (50) 

Histiocytic sarcoma 2 (4%) 3 (6%) 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 2 (4%) 3 (6%) 

.J 

3-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

(50) (50) 

I (2%) 

1 (2%) 

(50) (50) 

2 (4%) 4 (8%) 

3 (6%) 2 (4%) 

2 (4%) 

2 (4%) 1 (2%) 

(50) (5o) 

(22) (25), 

2 (8%) 

I (4%) 

(50) (50) 

2 (4%) 1 (2%) 

1 (2%) 

(49) (49) 

1 (2%) 

(50) (50) 

5 (lO%) 5 (1o%) " 

5 (lO%) 4 (8%) 

9 (18%) 9 (18%) 
349 

.... ~ ~ ~-.~:_~ ~ . ~ 

Dietary Restriction, N~?P T~ 460 

TABLE H~.b 

Summary of the Incidence of Neoplasms ~n Female IV~ice in the Dietary Restriction Study 

o~' Scopolamine Hydrobrornide Tr~hydrate: 2-~fear and 3-Yearr Restricted Feed ]?rotoeoHs (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

Neoplasm Summary 

Total animals with primary neoplasms c 

2-Year and 3-year protocols 

Total primary neoplasms 

2-Year and 3-year protocols 

Total animals with benign neoplasms 

2-Year and 3-year protocol s 

Total benign neoplasms 

2-Year and 3-year protocols 

Total animals with malignant neoplasms 

2-Year and 3-year protocols 

Total malignant neoplasms 

2-Year and 3-year protocols 

Total animals with metastatic neoplasms 

2-Year and 3-year protocols 

Total metastatic neoplasms 

2-Year and 3-year protocols 

Total animals with uncertain neoplasmsbenign or malignant 

3-Year protocol 

Total uncertain neoplasms 

3-Year protocol 

19 

20 

12 

12 

8 

8 

1 

2 

16 42 41 

17 68 65 

10 22 18 

10 28 29 

7 35 31 

7 39 36 

1 4 3 

4 8 6 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

b Number of animals with any tissue examined microscopically 

e Primary neoplasms: all neoplasms except metastatic neoplasms 
350 Dietary Restriction, NTP TR 460 

TA~L~ E[2a 

Statistical Analysis of Primary Neoplasms in Female Mice in the DietaryRestriction Study -. 

of Scopolamine Hydrobromide Trihydrate: 

i 

Harderian Gland: Adenoma 

Overall rate a 

Adjusted rateb 

Terminal ratec 

First incidence (days) 

Life table test d 

Logistic regression test d 

Fisher exact test d 

Harderian Gland: Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

•Fisher exact test 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate: 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Ad Libitum Feeding and. Weight-Matched Controls Protocols 

25 mg/kg x 25 mg/kg × 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

1/51 (2%) 1/51 (2%) 3/50 (6%) 1/51 (2%) 

7.7% 2.6% 

1/36 (3%) 1/38 (3%) 

707 726 (T) 

P=0.305N 

P=0.316N 

P=0.301N 

• , 

3/51 (6%) 1/51 (2%) 3/50 (6%) 1/51 (2%) 

7.2% 2.6% 7.7% 2.6% 

1/33 (3%) 1/38 (3%) 1/36 (3%) 1/38 (3%) 

511 726 (T) 707 726 (T) 

P=0.285N Pffi0.305N 

Pffi0.320N P=0.316N 

Pf0.309N P=0.301N , 

15/51 (29%) 6/51 (12%) 7150 (14%) 6/51 (12%) 

42.3% 15.4% 18.9% 15.4% 

13/33 (39%) 5/38 (13%) 6/36 (17%) 5/38 (13%) 

604 694 721 694 

P=0.011N P=0.469N 

P=0.017N P=0.526N 

P=0.024N P=0.485N 

8/51 (16%) 4/51 (8%) 2/50 (4%) 4/51 (8%) 

21.1% 10.3% 4.8% 10.3% 

5/33 (15%) 3/38 (8%) 1/36 (3%) 3/38 (8%) 

594 694 611 694 

P=0.139N P=0.345 

P=0.170N P=0.339 

P =0.179N P=0.348 

22/51 (43%) 9/51 (18%) 9/50 (18%) 9/51 (18%) 

57.1% 23.1% 23.2% 23.1% 

17/33 (52%) 8/38 (21%) 7/36 (19%) 8/38 (21%) 

594 694 611 6~ 

P=0.002N P=0.568N 

P=0.003N P=0.578 

P=0.005N P=0.584N 

• " 
351 

....... f ~, 

Dieh~ry Restriction, NT~ T]~ ~ 

TABLE ][-]I2a 

Stat~sfica~ Ana]ys~ of ]?rhnary Neop~mn~s ~n ~¢en~a~e Mice ~n the Dietary Rest~cfion Study 

o~ Scopo~anfine ~ydrobrom~de Tr~hydrate: Ad Libigum ~'eed~ng and We~ght-MatchedI Controh ~?r'otocoLs (continued) 

25 mg/kg x 25 mg/kg x 

Ad Libitum- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

Lung: Aiveolar/bronchiolar Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Ovary: Cystadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

PituRary Gland (Pars Distalis): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Thyroid Gland (Follicular Cell): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

3/51 (6%) 

8.0% 

1/33 (3%) 

618 

4/51 (8%) 

10.8% 

2/33 (6%) 

618 

3/51 (6%) 

7.1% 

0/33 (0%) 

511 

4/50 (8%) 

II.8% 

3/33 (9%) 

72O 

3/51 (6%) 

9.1% 

3/33 (9%) 

726 (T) 

2/51 (4%) 

5.3% 

2138 (5 %) 

726 (T) 

P=0,453N 

P=0.493N 

P=0.500N 

3/51 (6%) 

7.4% 

2/38 (5%) 

6O4 

P=0.450N 

P =0.496N 

P=0.500N 

1/51 (2%) 

2.6% 

1/38 (3%) 

726 (T) 

P=0.289N 

P=0.316N 

P=0.309N 

3/46 (7%) 

8.2% 

2/33 (6%) 

670 

Pffi0.484N 

P--0.543N 

P=0.547N f 

2/51 (4%) 

4.9% 

1/38 (3%) 

677 

P=0.442N 

P=0.483N 

P=0.500N 

2150 (4%) 

3/50 (6%) 

7.8% 

2/36 (6%) 

7O7 

0/49 (0 %) 

1/48 (2%) 

2.2% 

0/34 (0%) 

617 

1/50 (2%) 

2151 (4%) 

3/51 (6%) 

7.4% 

2/38 (5%) 

604 

P=0.655N 

P=0.661N 

P=0.652N 

1/51 (2%) 

3/46 (7%) 

8.2% 

2/33 (6%) 

670 

P=0.299 

P=0.290 

P=0.292 

2/51 (4%) 
352 Dietary Restriction, NTP TR 460 

TABIJ~ H2a 

Statistical Analysis of Primary Neoplasms in Female Mice in the Dietary Restriction Study ~ 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued~ 

Ad LibitumFed Control 

25 mg/kg x 

Ad ILibitumFed Control 

Weight-Matched 

Control 

25 mg/kg x 

Weight-Matched 

Control 

All Organs: Hemangiosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

4/51 (8%) 

11,3% 

2/33 (6%) 

709 

I151 (2%) 

2.6% 

1/38 (3%) 

726 (T) 

P=0.151N 

P=0.162N 

P=0.181N 

3/50 (6%) 

8.0% 

2/36 (6%) 

721 

1/51 (2%) 

2.6% 

1/38(3%) 

726 (T) 

P=0.292N 

P=0.316N 

P=0.301N 

All Organs: Histiocytic Sarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table te~,;t 

Logistic regression test 

Fisher exact test 

1/51 (2%) 1/51 (2%) 4/50 (8%) 

8.8% 

0/36 (0%) 

589 

1/51 (2%) 

2.5% 

0/38 (0%) 

691 

P=0.206N 

P=0.172N 

P=0.175N 

All Organs: Malignant Lymphoma (Lymphocytic or Mixed) 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

9/51 (18%) 

23.0% 

5/33 (15%) 

581 

7/51 (14%) 

16.2% 

3/38 (8%) 

551 

P=0.332N 

P=0.393N 

P=0,393N 

6/50 (12%) 

15,6% 

5/36 (14%) 

464 

7/51 (14%) 

16.2% 

3/38 (8%) 

551 

P=0.517 

P=0.519 

P=0.515 

All Organs: Benign Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

24/51 (47%) 

60.9% 

18/33 (55%) 

511 

18/51 (35%) 

43.8% 

15/38 (39%) 

670 

P=O.o72N 

P=O. 129N 

P=0.157N 

14/50 (28 %) 

34.7% 

10/36 (28%) 

617 

18/51 (35%) 

43.8% 

15/38 (39%) 

670 

P=0.309 

P=0.225 

P=0.283 

All Organs: Malignant Neoplasms 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

26/51 (51%) 

54.8% 

12/33 (36%) 

492 

14/51 (27%) 

31.6% 

8/38 (21%) 

551 

P=0.016N 

P=0.025N 

P=0.013N 

16/50 (32 %) 

36.2% 

9/36 (25 %) 

464 

14/51 (27%) 

31.6% 

8/38 (21%) 

551 

P=0.410N. 

P=O.405N 

P=0.389N 
I 

I 

J~ 

Die~ry Restriction, NT~ TR 460 353

TABLE ~a 

StatLstica~ AnagysLs of ]Primary Neopnasmsin Female Mice in the Dietary Restr~ct~or~ Study 

o~' Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-Matched ControLs ~'rotocoLs (continued) 

25 mg/kg x 25 mg/kg x 

Ad Libitura- Ad Libitum- Weight-Matched Weight-Matched 

Fed Control Fed Control Control Control 

All Organs: Benign or Malignant Neoplasms 

Overall rate 40151 (78%) 29/51 (57%) 28•50 (56%) 29/51 (57%) 

Adjusted rate 83.2% 64.3% 60.5% 64.3% 

Terminal rate 25/33 (76%) 22/38 (58%) 18/36 (50%) 22/38 (58%) 

First incidence (days) 492 551 464 551 

Life table test P=0.013N P=0.551 

Logistic regression test P=0.029N P=0.467 

Fisher exact test P=0.017N P=0.545 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for liver, lung, ovary, 

pituitary gland, and thyroid gland; for other tissues, denominator is number of animals necropsied. b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality ¢ Observed incidence at terminal kill 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the ad libitum or weight-matched controls and the dosed 

group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic 

regression test regards these lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in the 

dose group is indicated by N. 

Not applicable; no neoplasms in animal group 
-- -- 

354 Dietary Restriction, NTP TR 460 

TABLE H2b 

Statistical Analysis of Primary Neoplasms in Female Mice in the Dietary Restriction Study, 

of Scopolamine Hydrobromide Trihydrate: 2-YeaP and 3-Year Restricted Feed Protocols. 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

Bone Marrow: Hemangiosarcoma 

Overall ratea 0/50 (0%) 0/50 (0%) 

Adjusted rate b 

Terminal rate c 

First incidence (days) 

Life table test d 

Logistic regression test d 

Fisher exact test d~ 

Harderimt Gland: Adenoma 

Overall rate 3/50 (6%) O/50 (0 %) 

Adjusted rate 6.4% 0.0% 

Terminal rate 3/47 (6%) 0/44 (0%) 

First incidence (days) 723 (T) 

Life table test P=0.133N 

Logistic regression test P=0.133N 

Fisher exact test P=0.121N 

Liver: Hepatocellular Adenoma 

Overall rate 3/50 (6%) 3/50 (6%) 

Adjusted rate 6.4% 6.8% 

Terminal rate 3/47 (6%) 3/44 (7%) 

First incidence (days) 723 (T) 723 (T) 

Life table test P=0.632 

Logistic regression test P=0.632 

Fisher exact test P=0.661N 

Liver: Hepatocellular Carcinoma 

Overall rate: 0/50 (0%) 0/50 (0%) 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rate: 3•50 (6%) 3/50 (6%) 

Adjusted rate 6.4 % 6.8 % 

Terminal rate 3/47 (6 %) 3/44 (7 % ) 

First incidence (days) 723 (T) 723 (T) 

Life table test P=0.632 

Logistic regression test P=0,632 

Fisher exact test P=0.661N • 

3/50 (6%) 

7.1% 

0/20 (0%) 

611 

0/50 (0%) 

0.0% 

0/20 (O%) 

12/50 (24%) 

47.5% 

8/20 (40%) 

771 

4/50 (8%) 

13.4% 

1/20 (5%) 

901 

15/50 (30%) 

54.8% 

9/20 (45%) 

771 

1/50 (2%) 

5.3% 

1/19 (5%) 

1,087 (T) 

P=0.318N 

P=0.281N 

P=0.309N 

3/50 (6%) 

11.5% 

1/19 (5%) 

972 

P=O.r 140 

P=O.121 

P=O.121 

8/50 (16%) 

31.8% 

3/19 (16%) 

987 

P=0.241N 

P=0.207N 

P=0.227N 

4•50 (8%) 

15.6% 

2/19 (11%) 

848 

P=0.639N 

P--0.643 

P=0.643N 

I 1/50 (22%) 

39.6% 

4/19 (21%) 

848 

P=0.266N 

P--0.232N 

P=0.247N 

 ̧%%, ̧ 
Di.etary Restriction, N'~ Tlg 460 355 

TA~L~ ]HI2b 

Stat~st~ca~ Analysis of ~r~mary Heo#m~ns ~a ~ema~e I~ce ~n the D~eta~y ~estr~ct~on Study } off' Scop~aaa~ne ~ydrebrom~e Trr~hydrate: 2-Year and 3-Year ~estr~cte~ Feed ]?rotocogs (continued) 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

Lung: Alveolar/bronchiolar Adenoma 

Overall rate 3•50 (6%) 2/50 (4%) 

Adjusted rate 6.4% 4.4% 

Terminal rate 3/47 (6%) 1/44 (2%) 

First incidence (days) 723 (T) 666 

Life table test P=0.528N 

Logistic regression test P=0.508N 

Fisher exact test P=0.500N 

Lung: Alveolar/bronchiolar Carcinoma 

Overall rate 3150 (6%) 0150 (0%) 

Adjusted rate 6.3% 0.0% 

Terminal rate 2/47 (4%) 0/44 (0%) 

First incidence (days) 628 

Life table test P--0.134N 

Logistic regression test P=0.107N 

Fisher exact test P=0.121N 

Lung: Aiveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 6/50 (12%) 2150 (4%) 

Adjusted rate 12.5 % 4.4% 

Terminal rate 5/47 (11%) 1/44 (2%) 

First incidence (days) 628 666 

Life table test P=0.159N 

Logistic regression test P=0.129N 

Fisher exact test P=0.134N 

Pancreatic Blets: Adenoma or Carcinoma 

Overall rate 0/50 (0%) 0/50 (0%) 

.~, Adjusted rate 

] 

i

i 

]i 

Terminal rate 

First incidence (days) 

Life table test : ~ 

:]:: l 

:i:::: l l 

Logistic regression test 

Fisher exact test 

~: •[ Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 0/46 (0%) 1/46 (2%) 

Adjusted rate 

~[ Terminal rate 

First incidence (days) 

Life table test 

: ] 

~-::: [ Logistic regression test 

Fisher exact test 

i 

3-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

2/50 (4%) 

6.9% 

1/20 (5%) 

555 

3150 (6%) 

8.9% 

0/20 (0%) 

860 

5/50 (10%) 

15.2% 

1/20 (5%) 

555 

3/49 (6%) 

11.5% 

1/20 (5%) 

898 

3/42 (7%) 

16.7% 

3/18 (17%) 

1,087 (T) 

4/50 (8%) 

15.4% 

1/19 (5%) 

848 

P=0.332 

P=0.338 

P--0.339 

2/50 (4%) 

10.5%. 

2/19 (11%) 

1,087 (T) 

P=0.528N 

P=0.500N 

P=0.500N 

6/50 (12%) 

24.8% 

3/19 (16%) 

848 

P=0.474 

P =0.499 

P=0.500 

2/50 (4%) 

6.6% 

0/19(0%) 

703 

P=0.504N 

P=0.491N 

P=0.490N 

2/49 (4%) 

8.0% 

0/19 (0%) 

987 

P=0.471N 

P=0.425N 

P=0.427N 

...... ~ 

.... , 

~= I 
356 Dietary Restriction, NTP TR 460 

TABLE H2b 

Statistic.al Analysis of Primary Neoplasms in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control ~ 25 mg/kg 

Pituitary Gland (Pars Intermedia): Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Skin (Subcutaneous Tissue): 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

Spleen: ltemangiosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

All Orgaus: Hemangiosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Fisher exact test 

1/46 (2%) 0/46 (0%) 4/42 (10%) 2•49 (4%) 

16.5% 7.3% 

1/18 (6%) 0/19 (0%) 

946 980 

P=0.315N 

P=0.086N 

P=0.268N 

Sarcoma 

0/50 (0%) 0/50 (O%) 4/50 (8%) 5/50 (10%) 

11.0% 13.5% 

0/20 (0%) 0/19 (0%) 

537 737 

P=0.502 

P=0.537 

P=0.500 

Fibrosarcoma or Sarcoma 

0/50 (0%) 0/50 (0%) 5/50 (10%) 5/50 (10%) 

14.4% 13.5% 

0/20 (0%) 0/19 (0%) 

537 737 

P=0.615N 

P=0.603N 

P=0.630N 

0/50 (0%) 0/50 (o%) 1/50 (2%) 3/50 (6%) 

2.1% 13.2% 

0/20 (0%) 2/19 (11%) 

611 952 

P--0.318 

P=0.306 

P=0.309 

0/50 (0%) 0/50 (0%) 3/50 (6 %) 3/50 (6 %) 

7.1% 13.2% 

0/20 (0%) 2/19 (11%) 

611 952 

P=0.656N 

P=0.656N 

P=0.661N 
Dietary Restriction, NTP TR 460 357 

TABLE H2b 

S~at~sticzH Analysis of ]?rhnary Neoplasms in Femz~e I~,~ice in the D~et~ry ]Restriction Study. - i 

of Scopolamine Hydrobromlde Trihydrate: 2-~esr and 3-~ezr Restricted Feed ]Protocols (continued) 

2-Year Restricted Yeed ~3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg - i 

, All Organs: Histiocytic Sarcoma 

:~ : I Overall rate 2/50 (4%) 3/50 (6%) 5/50 (10%) 5/50 (10% 

-. Adjusted rate 4.1% 6.4% 13.9% 16.0% 

~, Terminal rate 1/47 (2%) 1/44 (2%) 1/20 (5%) 1/19 (5%) 

597 555 679 " LifeFirsttableincidencetest(days) 492 P=0.481 P=0.615N 

Logistic regression test P=0.532 P=0.609N 

~:: Fisher exact test P=0.500 P=0.630N : ~ 

~: 

:~ All Organs: Malignant Lymphoma (Lymphocytic or Mixed) 

[ Overall rate 2/50 (4%) 3/50 (6 %) 14/50 (28 %) 13/50 (26%) 

: Adjusted rate 4.3% 6.8% 47.0% 35.1% 

? l Terminal rate 2/47 (4%) 3/44 (7 % ) 6/20 (30 %) 1/19 (5%) 

i First incidence (days) 723 (T) 723 (T) 771 561 

!i i Life table test P=0.470 P=0.518N 

P=0.470 P=0.499N i FisherL°gistiCexactregressi°ntest tt P=0.500 P=0.500N 

~ ~1 All Organs: Benign Neoplasms 

~ [ Overall rate 12/50 (24%) 10/50 (20%) 22/50 (44%) 18/50 (36%) 

~' Adjusted rate 25.5 % 22.2% 71.2% 60.2% 

l Terminal rate 12/47 (26%) 9/44 (20%) 12/20 (60%) 8/19 (42%) 

"~ l ! 

First incidence (days) 723 (T) 666 555 703 

' Life table test P=0.473N P=0.301N 

Logistic regression test P=0.452N P=0.259N • . 

i Fisher exact test P=0.405N P=0.270N 

l 

[ All Organs: Malignant Neoplasms 

[ Overall rate 8/50 (16%) 7/50 (14%) 35/50 (70%) 31/50 (62%) 

I Adjusted rate 16.0% 14.8% 75.2% 71.4% 

~ [ Terminal rate 5/47 (11%) 4/44 (9 %) 9/20 (45 %) 8/19 (42%) 

[ First incidence (days) 492 538 537 561 

Life table test P=0.541N P=0.364N 

. P=0.429N 

~ : I Fisher exact test P=0.500N P=0.263N 

• Logistic regression test P=0.261N 

I : : I,

! 
358 Dietary Restriction, NTP TR 460 

TABLE H2b 

Statistical Analysis of Primary Neoplasms in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

All Organs: Benign or Malignant Neoplasms 

Overall rate 19/50 (38 % ) 16/50 (32 %) 42/50 (84%) 41/50 (82%) 

Adjusted rate 38.0% 33.3 % 87.2% 88.9% 

Terminal rate 16/47 (34%) 12/44 (27%) 14/20 (70%) 14/19 (74%) 

First incidence (days) 492 538 537 561 

Life table test P=0.426N P=0.518N 

Logistic regression test P=0.285N P=0.600N 

Fisher exact test P=0.338N • P=0.500N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for bone marrow, liver, 

lung, pancreatic islets, pituitary gland, and spleen; for other tissues, denominator is number of animals necropsied. 

b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

¢ Observed incidence at terminal kill 

d Beneath the dosed group incidence are the P values corresponding to pairwise comparisons between the controls and that dosed group. The life table test 

regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test regards these 

lesions as nonfatal. The Fisher exact test compares directly the overall incidence rates. For all tests, a lower incidence in the dose group is indicated by N. 

¢ Not applicable; no neoplasms in animal group 
Dietary Restriction, NTP TR 4(~, 359 

TABLE ~][3a 

Summary of the Incidence of Nonneoplastic Lesions in ~'emale 1V~lJce in the Dietary Restriction Study 

of Scopolamine H y d r o b r o m i d e T r i h y d r a t e : Ad Libitum ~eeding and Weight-Matched Controls ~'rotoco~s a 

Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

Disposition Summary 

Animals initially in study 70 b 60 70 b 

15-Month interim evaluation 10 10 10 

Early deaths 

Accidental deaths 2 1 

Moribund 9 10 7 

Natural deaths 7 3 6 

Survivors 

Terminal sacrifice 33 36 38 

Animals examined microscopically 61 60 61 

. 15-Month Ynterim Evaluation 

~ [ Alimentary System 

Liver (10) (10) (10) 

i Basophilic focus 1 (10%) 

. ] Inflammation 1 (10%) . 

:~ i I

I 

Mixed cell focus 

Necrosis 

1 (10%) 

1 (10%) 

Pancreas (10) (10) (10) 

= Atrophy 1 (10%) 

• Stomach, forestomaeh (10) (10) (10) 

Hyperplasia, focal 2 (20%) I (10%) 2 (20%) 

i 

l Endocrine System 

:~ i Adrenal cortex (10) (10) (10) 

~ ~ . Accessory adrenal cortical nodule 1 (10%) 

: = i Parathyroid gland (10) (10) (8) 

.... I Cyst 1 (10%) 

Pituitary gland (10) (io~ (1o) ~ ~ Pars distalis, hyperplasia 3 (30%) I (10%) 

~ Thyroid gland (10) (10) (10) 

i Follicular cell, hyperplasia 2 (20%) 

i ! Inflammation, chronic active 1 (10%) 

~ i - i i ~ [ 

Genital System 

Ovary (10) (10) (10) ~:~: i

I Cyst I (10%) I (10%) 2 (20%) 

I 

:i I 

:1 

Uterus 

Hyperplasia, cystic 

(1o) 

5 (50%) 

(1o) 

1 (10%) 

(1o) 

4 (40%) 

i ! 

a Number of animals examined microscopically at the site and the number of animals with lesion 

b Nine animals were removed for supplemental evaluations that were not included in the dietary restriction study. 

I 
360 Dietary Restriction, NTP TR 460 

TABLE lt3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobr0mide Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

• 

• 

- Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

15-Month Interim Evaluation (continued) 

Hematopoietic System 

Bone marrow (10) (I0) (10) 

Myelofibrosis 

Thymus 

1 (10%) 

(9) 

1 (10%) 

(10) (10) 

Inflammation, chronic active . 

• 

1 (10%) 

Mineralization 1 (11%) 

Respiratory System 

Lung (10) (10) (I0) 

Inflammation, chronic active 1 (10%) 

Alveolar epithelium, hyperplasia 1 (10%) 

Urina~y...System 

Kidney ( 1 O) (I0) (lO) 

Nephropatthy-.. 5 (50%) 2 (20%) 2 (20%) 

Systems Examined With No Lesions ~Observed 

Cardiovascular System 

General Body System 

Integumentary System 

Musculoskeletal System 

Neryous System 

Spedal~Se.nses System 

2-Year Study 

Alimentary System 

Esophagus (51) (50) (50) 

Perforation 1 (2%) 1 (2%) 

Periesophageal tissue, inflammation, 

suppurative 3 (6%) 1 (2%) 

Intestine large, rectum (51) (50) (50) 

Inflammation, chronic active 1 (2%) 

Intestine small, jejunum (51) (49) (50) 

Hyperplasia, lymphoid 2 (4%) 2 (4%) 3 (6%) 

Inflammation, chronic active 1 (2%) 

Liver ~ (51) (50) (51) 

Angiectasi~.. 1 (2%) 2 (4%) 1 (2%) 

Basophilic, focus 1 (2%) 3 (6%) 

Clear cell focus (2%) 

Ectopic tissue (2%) 

Eosinophilic focus 17 (33%) (8%) 9 (18%) 

Fatty change (2%) 

Hematopoietic cell proliferation (2%) 1 (2%) 
Dietary Restriction, NTP TR 460 361 

TABLE H3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the Dietary lRestriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitura Feeding and Weight-Matched Controls ~'rotocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 25 mg/kg 

2-Year Study (continued) 

Alimentary System (continued) 

Liver (continued) (51) (50) (51) 

Inflammation, chronic active 1 (2%) 

Mixed cell focus 6 (12%) 1 (2%) 3 (6%) 

Necrosis 3 (6%) 1 (2%) 

Mesentery (7) (5) (5) 

Inflammation, suppurative 2 (40%) 

Fat, necrosis 6 (86%) 1 (20%) 2 (40%) 

Pancreas (51 ) (50) (51 ) 

Atrophy 5 (10%) 4 (8%) 

Hypertrophy 1 (2%) 

Duct, cyst 2 (4%) 3 (6%) 

Salivary glands (51) (50) (51) 

Atrophy 1 (2%) 

Duct, hyperplasia 1 (2%) 

Stomach, forestomach (51) (50) (5!) 

Cyst 1 (2%) 

Diverticulum 1 (2%) 

Hyperplasia, focal 6 (12%) 16 (32%) 12 (24%) 

Stomach, glandular (51 ) (50) (51 ) 

Erosion 2 (4%) 2 (4%) 

Mineralization 1 (2%) 

Cardiovascular System 

Blood vessel (51) (50) (51) 

Aorta, inflammation, chronic active 1 (2%) 

Mineralization 1 (2%) 

Heart (51) (50) (51) 

Inflammation, chronic active 1 (2 %) 

Mineralization 2 (4%) 1 (2%) 

Artery, inflammation, chronic active 2 (4%) 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

(51) (50) 

2 (4%) 

(51) 

1 (2%) 

Hyperplasia 1 (2%) 1 (2%) 

Adrenal medulla (51) (49) (50) 

Hyperplasia 3 (6%) 1 (2%) 

Islets, pancreatic (51) (50) (51) 

Hyperplasia 1 (2%) 1 (2%) 1 (2%) 

Parathyroid gland (48) (38) (45) 

Cytoplasmic alteration, focal 1 (3%) 

Pituitary gland (50) (48) (46) 

Pars distalis, hyperplasia 24 (48 %) 11 (23%) 13 (28%) 

Pars intermedia, hyperplasia 1 (2%) 

Thyroid gland (51) (50) (51) 

Inflammation, chronic active 1 (2%) 1 (2%) 

Follicular cell, hyperplasia 16 (31%) 5 (10%) 8 (16%) 
362 Dietary Restriction, NTP TR 460 

TABLE H3a 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) .~ 

2-Year Study (continued) 

General Body System 

None 

Genital System 

Clitoral gland 

Duct,. ectasia 

Ovary 

Cyst 

Inflammation, suppurative 

Uterus 

H~per~iasia, cystic 

Inflammation, chronic active 

Thrombosis 

Hematopoietic System " 

Bone marrow 

Myelofibrosis 

Erythroid cell, hyperplasia 

Myeloid cell, hyperplasia 

Lymph node 

Bronchial, inflammation, Chronic active 

Lymphnode, mandibular 

Hyperplasia, lymphoid 

Lymph node;, mesenteric 

Hyperplasia, lymphoid 

Inflammati~on, chronic active 

Spleen 

Depletion lymphoid 

Hematopoietic cell Pr01if.er~tion 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Thymus 

Atrophy 

Integumentary System 

Mammary gland 

Hyperplasia 

Skin 

Cyst epithelial inclusion 

Inflammation, chronic active 

Ulcer 

Ad LibitumFed Control 

(48) 

(51) 

16 (31%) 

(51) 

38 (75%) 

1 (2%) 

(51) 

22 (43%) 

3 (6%) 

9 (18%) 

•(6) 

(5o) 

1 (2%) 

(50) 

(51) 

2 (4%) 

17 (33%) 

3 (6%) 

1 (2%) 

(48) 

3 (6%) 

(51) 

I (2%) 

(51) 

1 (2%) 

I (2%) 

Weight-Matched 

Control " 

(50) 

1 (2%) 

(49) 

9 (18%) 

2 (4%) 

(50) 

28 (56%) 

1 (2%) 

1 (2%) 

(50) 

17 (34%) 

2 (4%) 

7 (14%) 

(3) 

1 (33%) 

(48) 

(48) 

I (2%) " 

1 (2%) 

(50) 

I (2%) 

II (22%) 

2 (4%). 

(43) 

2 (5%) 

(5O) 

(50) 

25 mg/kg 

(48) 

1 (2%) 

(51) 

12 (24%) 

I (2%) 

(51) 

23 (45%) 

(51) 

13 (25%) 

3 (6%) 

(3) 

(51) 

1 (2%) 

(48) 

2 (4%) 

(51) 

7 (14%) 

2 (4%) 

(45) 

3 (7%) 

(51) 

4 (8%) 

(51) ~ 

i (2%) 
~:~ ~--~-~-~:-~-~.~ :~:~ ~ 

Dietary Restriction, NT]? Tg ~60 3f~33 

TABLE ~]I3 a 

Sun~n~ary of the Incidence of Nonneoplastic Lesions in I~emale I~ice in the Dietary Restriction Study 

of Scopoimnine Hydrobro~nide Trihydrate: -Ad Libitum Feeding and Weight-Matched Controas ~rotoco~s, continued) 

~ i 

~: I ~ ~ ~ [ 

_ ; 

~ i ! 

= I 

' ~ > I 

: i 

ii~ 

i:i 

~ I 

~~:i ~ 

i Ii 

: i :?: ! 

;:: 1 

, 

2-Year Study (continued) 

Musculoskeletal System 

None 

Nervous System 

Brain 

Infarct 

Neuron, necrosis 

Peripheral nerve 

Degeneration 

Respiratory System 

Lung 

Inflammation, chronic active 

Special Senses System 

Eye 

Cornea, inflammation, chronic active 

Ear 

Internal ear, inflammation, chronic active 

Urinary System 

Kidney 

Cyst 

Infarct 

Inflammation, chronic active 

Mineralization 

Nephropathy 

Artery, inflammation, chronic active 

Renal tubule, necrosis, acute 

Urimry bladder 

Inflammation, chronic active 

Artery, inflammation, chronic active 

Ad LibitumFed Control 

(51) 

1 (2%) 

1 (2%) 

(2) 

2 (100%) 

(51) 

1 (2%) 

(2) 

2 (100%) 

(51) 

I (2%) 

1 (2%) 

23 (45%) 

I (2%) 

(50) 

Weight-Matched 

Control 

(50) 

4 (8%) 

(2) 

1 (50%) 

(50) 

(1) 

1 (100%) 

(50) 

1 (2%) 

23 (46%) 

(50) 

25 mg/kg 

(51) 

2 (4%) 

(51) 

(51) 

' 

1 (2%) 

10 (20%) 

l (2%) 

(5 I) 

I (2%) 

I. (2%) 
364 Dietary Restriction, NTP TR 460 

TABL:E H3b 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols a ' 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

Disposition Summary 

Animals initially in study 60 60 50 50 

15-Month interiin evaluation 10 10 

Early deaths 

Accidental deaths 1 1 

MOribund 2 2 18 16 

Natural deaths 1 3 12, 14 

Survivors 

Terminal sacrifice 47 44 20 19  ̧ 

Animals examined microscopically 60 60 50 50 

15.Month Interim Evaluation 

Alimentary System 

Liver (10) (10) 

Basophilic focus 1 (10%) 

Stomach, forestumach (10) (10) 

•Hyperplasia, focal 7 (70%) 6 (60%) 

Cardiovascular System 

Heart • (I0) (10) 

Inflammation, chronic active 1 (10%) 

Endocrine System 

Adrenal cortex (10) (10) 

Accessory adrenal cortical nodule 1 (10%) 

Parathyroid gland (8) (8) 

Cyst 1 (13%) 

Pituifary gland (10) (I0) 

Pars distalis, hyperplasia 1 (10%) 

Genital System 

Ovary (10) (10) 

Cyst 1 (10%) 

Uterus (10) (10) 

Hyperplasia, cystic 2 (20%) 

Respiratory System 

Lung (10) (10) 

Inflammation, chronic active ! (10%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 

J 

/ 

/ 
:~ ~: 

! 

Die~ry Restriction, NTP TR 460 365 i 

TABLE H3b 

Summary of the ~ncidence of Nonneopl~stic Lesions in Female IV~ice in the Dietary Restriction Study 

of Seopolmnine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

i 

2-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

3-Year Restricted Feed 

Vehicle 

Control 25 mg/kg 

15-Month Interim Evaluation (continued) 

Special Senses System 

Harderian gland (3) 

Inflammation, chronic active 1 (33%) 

Systems Examined With No Lesions Dbserved 

General Body System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Urinary System 

2-Year and 3-Year Protocols 

Alimentary System 

Esophagus (50) (50) (50) (50) 

Periesophageal tissue, degeneration 1 (2%) 

Periesophageal tissue, hemorrhage 1 (2%) 

Gallbladder (50) (50) (50) (49) 

Inflammation, chronic active 1 (2%) 1 (2%) 

Intestine small, jejunum (50) (50) (50) (49) 

Hemorrhage 1 (2%) 

Hyperplasia, lymphoid 1 (2%) 

Liver (50) (50) (50) (50) 

Angiectasis 1 (2%) 1 (2%) 

Basophilic focus 1 (2%) 1 (2%) 3 (6%) 

Clear cell focus 2 (4%) 

Eosinophilic focus 1 (2%) 4 (8%) 

Hematopoietic cell proliferation 1 (2%) 2 (4%) . 

Hyperplasia, lymphoid 1 (2%) 

Inflammation, focal 1 (2%) 

Mixed cell focus 2 (4%) 1 (2%) 

Necrosis 1 (2%) 2 (4%) 1 (2%) 

Bile duct, cyst 2 (4%) 

Bile duct, hyperplasia 1 (2%) 

Centrilobular, necrosis 1 (2%) 

Serosa, pigmentation, hemosiderin 1 (2%) 

Pancreas (50) (50) (49) (50) 

Acinus, atrophy 3 (6%) 4 (8%) 3 (6%) 

Acinus, hyperplasia, focal 3 (6%) 

Artery, inflammation, chronic active 2 (4%) 

Duct, ectasia 1 (2%) I (2%) 2 (4%) 
366 Dietary Restriction, NTP TR 460 

TABLE H3b 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control 25 mg/kg Control 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Alimentary System (continued) 

Stomach, forestomach 

Cyst 

(50) (50) (50) 

1 (2%) 

(50) 

Erosion 1 (2%) 

Hyperplasia, focal 

Infdtration cellular, mast cell 

29 (58%) 

1 (2%) 

31 (62%) 23 (46%) 29 (58%) 

Mineralization 1 (2%) 

Stomach, glandular (50) (50) (50) (50) 

Dysplasia 1 (2%) 

Erosion 1 (2%) 

Mineralization 1 (2%) 

Tooth (2) (2) 

Dysplasia 2 (ioo%) 1 (50%) 

Cardiovascular System 

Blood vessel (50) (50) (50) (50) 

Inflammation, chronic active I (2%) 

Aorta, inflammation 1 (2%) 

Aorta, thrombosis 2 (4%) 

Heart (50) (50) (50) (50) 

Degeneration 1 (2%) 

Mineralization 2 ~4%) 1 (2%) 

Endocrine System 

Adrenal cortex (49) (5O) (50) (50) 

Accessory adrenal cortical nodule 1 (2%) i (2%) 

Angiectasis 

Hyperplasia 2 (4%) 1 (2%) 

1 (2%) 

2 (4%) 

2 (4%) 

2 (4%) 

Hypertrophy I (2%) 

Capsule, hyperplasia 1 (2%) 

Adrenal medulla (49) (50) (49) (50) 

Hyperplasia 

Islets, pancreatic (50) (50) (49) 

1 (2%) 

(50) 

Hyperplasia 

Parathyroid gland 

Cyst 

Pituitary gland 

(44) 

(46) 

(41) 

1 (2%) 

(46) 

1 (2%) 

(45) , 

(42) 

(46) 

1 (2%) 

(49) 

Pars distalis, hyperplasia 1 (2%) 4 (9%) 8 (19%) 1 (2%) 

Pars intermedia~ hyperplasia 

Thyroid gland .. 

1 (2%) 

(50) (49) (50) (50) 

Inflammation, chronic active 

Follicular cell, hyperplasia 

1 (2%) 

2 (4%) 9 (18%) 2 (4%) 

General Body System 

None 
Dietary Restriction, NT~ T~ 46~ 367 

 ̧= 

=: 

I 

I 

TABLE H3b 

Sumrnzry of the Incidence of Nonneo~s~c Lesions ~n Fem~de~Mice In the Die~ryj gestr]c~on gaudy 

of Scopo~anfine Hydrobromide Tr~hydrate: 2-Year and 3-Year ~estr~cted Feed Protocols (continued) 

"'7 I 2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle Vehicle 

Control •25 mg/kg Control 25 mg/kg 

: :l 2-Year and 3-Year Protocols (continued) 

I Genital System 

:: l : , Clitoral gland (49) (49) (50) (48) 

: [ Inflammation, chronic active 1 (2%) 1 (2%) 

~ ~ i Duct, ectasia 2 (4%) 

-l Ovary (50) (50) (49) (49) 

-

~; ~I Angiectasis 

Atrophy 1 (2%) 

1 (2%) 2 (4%) 1 (2%) 

i 

: I Cyst 

Hemorrhage 

13 (26%) 

1 (2%) 

9 (18%) 18 (37%) 19 (39%) 

=: i

I 

Inflammation, chronic active 

Thrombosis 

2 (4%) 

1 (2%) 1 (2%) 

I Corpus luteum, hyperplasia 1 (2%) 

[

i 

- ~ = i 

Uterus 

Hemorrhage 

(50) 

16 (32%) 

(50) 

15 (30%) 

(49) 

1 (2%) 

9 (18%) 

(49) 

11 (22%) 

_ 

::2 . 1 

~ 

Inflammation, suppurative 

Thrombosis 

1 (2%) 

1 (2%) 

~ ; I 

~ I~ HematopoieticSystem 

:~ 

[ 

[

I 

Bone marrow 

Angiectasis 

(50) (50) (50) 

1 (2%) 

(50) 

Myelofibrosis 7 (14%) 13 (26%) 19 (38%) 13 (26%) 

I 

.........~, 

Erythroid cell, hyperplasia 

Myeloid cell, hyperplasia 1 (2%) 

1 (2%) 

1 (2%) 

4 

9 

(8%). 

(18%) 

1 (2%) 

7 (14%) 

Lymph node (13) (8) 

Hematopoietic cell proliferation 1 (13%) 

i~ ::! Hyperplasia, lymphoid 1 (8%) 

!: Lymph node, mandibular (50) (48) (45) (47) 

= I Hematopoietic cell proliferation 1 (2%) 

Hyperplasia, lymphoid 1 (2%) 1 (2%) 1 (2%) 

Lymph node, mesenteric (44) (50) (43) (48) 

:~:: 

~ : 

:: 

~ 

I

I 

Angiectasis 

Hyperplasia, lymphoid 1 (2%) 

3 (7%) I (2%) 

: I , 

== i 

Spleen 

Depletion lymphoid 

(50) (50) (50) (50) 

1 (2%) 

E I Hematopoietic cell proliferation 3 (6%) 7 (14%) 21 (42%) " 17 (34%) 

::: :~:: !! Hyperplasia, lymphoid 1 (2%) 2 (4%) 

i Thymus (45) (49) (37) (42) 

I Atrophy 1 (2%) 1 (2%) 9 (24%) 5 (12%) 

: ~': l

! Hyperplasia, lymphoid 2 (4%) 

~ ~ I 

Integumentary System 

Mammary gland (49) (49) 

Hyperplasia (50) (50) I (2%) 

Skin (50) (50) (50) (50) 

Subcutaneous tissue, inflammation, 

chronic active 1 (2%) 1 (2%) 
368 Dietary Restriction, NTP TR 460 

TABLE H3b 

Summary of the Incidence of Nonne0plastic Lesions in Female Mice in the Dietary Restriction Study 

of Scopollamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols (continued) 

2-Year Restricted Feed 3-Year Restricted Feed 

Vehicle - -.Vehicle 

.Control 25 mg/kg Control • 25 mg/kg 

2-Year and 3-Year Protocols (continued) 

Musculoskeletal System 

None ! 

Nervous System 

Brain (50) (50) (50) (50) 

Granuloma, focal 1 (2%) 

Neuron, necrosis 1 (2%) . 

Respiratory system 

Lung (50) (50) (50) (50) 

Infiltration cdlular, histiocyte 2 (4%) 

Infiltration cellular, lymphocyte 1 (2%) 

Inflammation, chronic active I (2%) 

Alveolar epithelium, hyperplasia ~ 1 (2%) 4 (8%) i 4 (8%) 

Nose (50) , (49) (5o) .,. (50) 

Infdtration cellular, mast dell 1 (2%) • 

Special Senses System 

Eye (1) (3) 

Lens, cataract 1 (100%) 3 (100%) 

Hhrderian gland (24) (23) (22) (25) 

Hyperplasia 1 (4%) 

• 

Urinary System 

Kidney (50) (50) (50) (50) 

Cytoplasmic alteration 1 (2%) 1 ,(2%) . 

• Infdtration cellular, lymphocyte 1 (2%) 

Nephropathy 16 (32%) 6 (12%) • 19 (38%) 13 (26%) 

Pelvis, inflammation, chronic active .1 (2%) 

Renal • : tubule, necrosis 1 (2%) 

Urinary bladder 0o) (50) . (49). ,. (49) 

Inflammation, chronic active 1 (2%) 1 (2%) 

., 

.. 
369 !: 

~ I 

;~i [ 

i 

I 

: l 

5 i 

= 

2 ! 

} 

~: ! 

~ ~ I 

: = i 

. 

?5 " 

r: 

i 

~i i = ~ 1 

I ? I 

;; I 

1 

: I 

? I 

!. 

= ~ 

! 

1 

~ I 

I 

:~ 

: I 

AN 

TABLE [la 

TABLE ~lb 

'~'ABLE ~a 

T~L~ ~.2b 

TABLE ~3a 

.,~ 

TABLE I3b 

TABLE ~4a 

TABLE ][4b 

TABLE 15a 

T~L~ I5b 

• 

E 

© GAHoWE GHToT©oB©II YoWE GHT AT @5 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: Ad Libitum Feeding 

and Weight-Matched Controls Protocols ................................. 370 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Butyl Benzyl Phthalate: 2-Year and 30-Month Restricted Feed ]Protocols ......... 371 

Organ Weights and Organ-Weight-to,Body-Weight Ratios for Rats 

at the 3-Month Interhn Evaluation in the Dietary Restrriction Study 

of t-Butylhydroquinone: Ad Libitum Feeding 

and Weight-Matched Controls Protocols ............................ , .... 372 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

at the 3-Month Interim Evaluation in the Dietary Restrriction Study 

of t-Butylhydroquinone: Restricted Feed Protocol ....................... ;... 373 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Male Rats 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Sallcylazosulfapyridine: Ad Libitum Feeding 

and Weight-Matched Controls Protocols ................................. 374 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Male Rats 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 30-Month Restricted Feed Protocols ......... 375 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Male Mice 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding 

and Weight-Matched Controls Protocols ................................. 376 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Male Mice 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Salicylazosulfapyridine: 2-Year and 3-Year Restricted Feed Protocols ........... 377 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice 

at the 15-Month Interim Evaluation in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding 

and Weight-Matched Controls Protocols ...... . ........................... 378 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice 

at the 1S-Month ~nterhn Evaluation in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 

2-~earr and 3-Year Restricted Feed Protocols .............................. 379 

J -.~ j 
370 Dietary Restriction, NTP TR 460 

TABLE lla 

Organ Weights mad Organ.Weight-to-Body-Weight Ratios for Rats at the 15~Month Interim Evaluation 

in the Dietary Restriction Study of Butyl Benzyl Phthalate: Ad Libitum Feeding 

and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Control 12,000 ppm 

Male 

10 10 10 .. 

Necropsy body wt 458 ± 10 397 ± 9 416 ± 8"*~, 

Epklidymis 

Absolute 0.428 ± 0.016 0.451 ± 0.032 0.433 ± 0.009 

Relative 0.93 ± 0.02 1.14 ± 0.08 1.04 ± 0.02** 

R. Kidney 

Absolute 1.608 ± 0.045 1.425 ± 0.043 1.706 ± 0.048~ 

Relative 3.52 ± 0.10 3.58 ± 0.03 4.10 ± 0.08**A, 

Liver 

Absolute 15.463 ± 0.524 12.350 ± 0.429 15.761 ± 0.483~ 

Relative 33.75 ± 0.92 31.08 ± 0.64 37.82 ::t: 0.78"*~ 

R. Testis 

Absolute 1.638 ± 0.122 1.640 ± 0.177 1.771 ± 0.165 

Relative 3.62 ± 0.36 4.15 ± 0.46 4.27 ± 0.42 

Ad Libitum- Weight-Matched 

Fed Control Control 24,000 ppm 

Female 

n 10 10 10 

Necropsy body wt 279 5:7 198:5:2 215 ± 4"*,~ 

R. Kidney 

Absolute 0.931:5:0.022 0.674 ± 0.017 0.878 ± 0.018A* 

Relative 3.34:5:0.07 3.40 ± 0.06 4.09 ± 0.12"*~ 

Liver 

Absolute 8.629 ± 0.189 5.122:5:0.123 8.377:5: 0.153~A 

Relative 31.01 ± 0.81 25.87 ± 0.52 38.96 ± 0.57"*A, 

** Significantly different (P~0.01) from the ad libitum-fed control group by Student's t-test 

Ab .Significantly different (P~0.01) from the weight-matched control group by Student's t-test a ,Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

rag organ weight/g body weight (mean ± standard error). 
Dietzry Restriction, NT~ TR 460 37~. 

TA~L~ I]b 

Drgan. Weights and Organ-Weight-to-body-Weight Ratios for ~ats ~t the ~5-I~anth ]Interhn Evaluation 

in the Dietary ~estr~ction .Study of ~utyH ~enzy~ Phtha~ate: 

2-Fear and 3O-Manth ]Restricted ~eed ][h'otoeo~s a 

0 ppm ~12,000 ppm 

Male 

I 

: I n 10 10 2~ ~ 

~ 7 I 

Epididymis 

Absolute 0.464 ± 0.007 0.458 ± 0.016 

~ ] Relative 1.22 ± 0.03 1.26 ± 0.06 :~i,= ~ 

=~ i R. Kidney 

~ I Absoluto 1.329 ± 0.0~7 1.488 ± 0.020"* I I Relative 3.49 ± 0.07 4.08 ± 0.05** 

Liver 

'; Absolute 11.546 ± 0.248 12.229 ± 0.317 

-~ [ ' Relative 30.26 ± 0.47 33.51 ± 0.67** 

I R. Testis 

? Absolute 1.552 ± 0.017 1.552 ± 0.166 

:~ ,1 Relative 4.07 5:0.07 4.25 ± 0.44 

7~, ] 

: ~ I 

I 0 ppm ~,000 ppm .~,: I 

: ~ Ii Female _ 

£( 

...... ] 

~ ] Necropsy body wt 235 ± 3 195 ± 4** 

~ I 

~ I] R. Kidney 

~ i Absolute 0.838 5:0.012 0.808 5:0.014 

~ I - Relative 3.57 ± 0.06 4.15 ± 0.07** 

,,or 

..... Absolute 7.241 ± 0.149 7.191 ± 0.162 1 Relative 30.82 5:0.57 36.85 5: 0.45** 

** Significantly different (P~0.01) from the~control group by Student's t-test 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean ± standard error). 

-.= I 
372 Dietary Restriction, NTP TR 460 

TABLE I2a 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats at the 3-Month Interim Evaluation 

in the Dietary Restriction Study of t-Butylhydroquinone: Ad Libitum Feeding 

and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Control 5,000 ppm 

n 10 10 10 

Male 

Necropsy body wt 355 ±7 329±3 341 ±6 

Epididymis 

Absolute 0.472 ± 0.020 0.454 ± 0.009 0.479 ± 0.014 

Relative 1.33 ± 0.07 1.38 ± 0.03 1.41 ± 0.04 

R. Kidney 

Absolute 1.389 ± 0.038 1.261 ± 0.021 1.385 5: 0.052, 

Relative 3.91 ± 0.06 3.84 ± 0.05 4.06 ± 0.10 

Liver 

Absolute 13.467 ± 0.435 12.424 ± 0.592 14.420 ± 0.451, 

Relative 37.88 ± 0.62 37.81 ± 1.77 42.26 ± 0.86**, 

R. Testis 

Absolute 1.489 ± 0.022 1.451 ± 0.015 1.485 ± 0.022 

Relative 4.21 ± 0.11 4.42 ± 0.04 4.36 ± 0.07 

Female 

Necropsy body wt 199 ± 2 180 ± 2 181 ± 2** 

R. Kidney 

Absolute 0.749 ± 0.013 0.684 ± 0.006 0.675 ± 0.015"* 

Relative 3.76 ± 0.07 3.80 ± 0.04 3.73 ± 0.07 

Liver 

Absolute 6.310 ± 0.150 5.589 ± 0.085 6.255 ± 0.096,, 

Reiative 31.70 ± 0.80 31.07 ± 0.47 34.61 ± 0.37**,, 

**. Significantly different (P~O.O1) from the ad libitum-fed comrol group by Student's t-test 

A Significantly different (P~0.05) from the weight-matched control group by Student's t-test 

,, P~0.01 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given 

as mg organ weight/g body weight (mean ± standard error). 
Dietary ]Restriction, NTP TR 460 373 

T~d~LE ~b 

Organ Weights and Organ-Weight-to-Body-Weight ]Ratios for ]Rats at the 3-Month ~uterhn Eva~uatlou 

in the Dietary ]Restriction Study of t-~uty~hydroqu~none: lRestr~cted Yeed ][hrotoco~ a 

0 ppm 5,000 ppm 

n 10 10 

Male 

Necropsy body wt 303 ± 4 297 ± 4 

Epididymis 

Absolute 0.461 ± 0.010 0.464 ± 0.012 

Relative 1.52 ± 0.02 1.56 ± 0.03 

R. Kidney 

Absolute 1.195 ± 0.025 1.269 5:0.029 

Relative 3.95 ± 0.07 4.28 5: 0.08** 

Liver 

Absolute 10.851 5:0.323 12.871 5: 0.145"* 

Relative 35.79 ± 0.75 43.44 5: 0.70** 

R. Testis 

Absolute 1.494 5:0.018 1.460 5:0.031 

Relative 4.94 5:0.07 4.92 5:0.08 

Female 

Necropsy body wt 174 5:3 162 5: 2** 

R. Kidney 

Absolute 0.645 5:0.014 0.624 ::t: 0.018 

Relative 3.70 5:0.04 3.84 ± 0.10 

Liver 

Absolute 5.577 5:0.528 6.004 ± 0.511 

Relative 31.80 ± 2.46 37.00::1:: 3.19 

** Significantly different (P~0.01) from the control group by Student's t-test 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean 5: standard error). 
3"74 Dietary Restriction, NTP TR 460 

TABLE 13a 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Male Rats . 

at: the 15-Month Interim Evaluation in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and WeightTMatched Controls Protocolsa 

Ad Libitum- Weight-Matched 

Fed Control Control 377.5 mg/kg 

n 10 I0 10 

Necropsy body wt 491 ± 8 477 ± 8 459 ± 8* 

R. Kidney 

Absolute 1.639 ± 0.061 1.569 ± 0.043 1.590 ± 0.029 

Relative 3.33 ± 0.10 3.29 ± 0.07 3.47 ::1:0.05 

Liver 

Absolute 16.660 ± 0.544 15.768:5:0.431 15.713 ± 0.370 

Relative 33.89 ± 0.922 33.06 ::1:0.85 34.24 ± 0.69 

Spleen 

Absolute 0.803 ± 0.026 0.738 ± 0.034 0,727 ± 0.015" 

Relative 1.63 ± 0.04 1.64 ± 0.07 1.58 ± 0.02 

L. Testis 

Absolute 1.641 ± 0.055 1.783 ± 0.111 1.842 ± 0.167 

Relative 3.34 ± 0.08 3.65 ± 0.25 4.01 ± 0.35 

R. Testis 

Absolute 1.559 ± 0.040 1.512 ± 0.047 1.743 ± 0,157 

Relative 3.17 ± 0.06 3.18 ± 0.13 3.79 ::t: 0.32 

Thyroid gland 

Absolute 0.025 ± 0.003 0.020 ± 0.003 0.022 ± 0.002 

Relative 0.05 ± 0.01 0.04 ± 0.01 0.05 ± 0.00 

* Significantly different (P~0.05) from the ad libitum-fed control group by Student's t-test 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean 5: standard error). 
Dietary ll~estriction, N'~ T~ 460 37~ 

TA~L~ lI3b 

Organ Weights and Organ-Weight-toql~ody-We~ght ~atios for M~e ~ 

at the IS-Month ~nte~ Ev~uat~on ~ the D~e~ ~r~ct~on Study o~ Sa~cy~ulfap~e: 

~-~ ~d 3~oath ~r~cte~ Yeed ~ot~o~ a 

Vehicle Control 337.5 mg/kg 

~ [ 

~ ] 

n 10 10 

Necropsy body wt 419 ± 11 396 ± 8 

R. Kidney 

Absolute 1.404. ± 0.037 1.394 ± 0.04.2 

Relative 3.36 5:0.07 3.51 ::1:0.05 

Liver 

Absolute 13.635 ± 0.331 13.083 ± 0.498 b 

Relative 32.67 + 0.79 32.59 + 1.06 b 

Spleen 

Absolute 0.734 + 0.038 0.653 4- 0.014 

Relative 1.76 ± 0.10 1.65 + 0.04 

L. Testis 

Absolute 1.699 + 0.068 1.801 + 0.129 

Relative 4.06 + 0.11 4.59 ± 0.41 

R. Testis 

Absolute 1.443 ± 0.205 1.576 ± 0.085 ii ii~1 Relative 3.41 ± 0.48 3.97 ± 0.21 

Thyroid gland 

Absolute 0.017 ± 0.001 0.017 ::1:0.001 

Relative 0.04 ± 0.00 0.04 ± 0.00 

Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean ± standard error). 

n=9 

~ I 
376 Dietary Restriction, NTP TR 460 

T~kBLE I4a 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Male Mice 

at the 15-Month Interim Evaluation in the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Control 2,700 mg/kg 

n 10 10 10 

Necropsy body wt 53.9 ± 1.1 45.7 ± 1.4 44.9 ± 1.3"* 

R. Kidney 

Absolute 0.424 ± 0.018 0.353 ± 0.010 0.359 ± 0.011"* 

Relative 7.87 ± 0.30 7.76 ± 0.22 8.05 ± 0.30 

Liver 

Absolute 2.328 ± 0.106 1.610 ± 0.112 2.305 ± 0.062~ 

Relative 43.02 ± 1.15 35.01 ± 1.59" 51.55 ± 1.19**~A 

Spleen 

Absolute 0.100 ± 0.008 0.073 ± 0.005 0.119 ± 0.006AA 

Relative 1.86 ± 0.17 ~ 1.61 ± 0.12 2.67 ± 0.15**A~ 

R. Testis 

Absolute 0.113 ± 0.004 b 0.117 ± 0.003 0.108 ± 0.005 

Relative 2.11 ± 0.08 b 2.57 ± 0.07 2.44 ± 0.14 

Thyroid gland 

Absolute 0.003 ± 0.000 0.004 ± 0.000 0~004 ± 0.000 

Relative 0.06 ± 0.01 0.08 ± 0.01 0.08 5:0.01 

** Significantly different (P~0.01) from the ad libitum-fed control group by Student's t-test 

AA Significantly different (P~0.01) from the weight-matched control group by Student's t-test 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are g~ven as 

mg organ weight/g body weight (mean ± standard error). 

b n=9 
Dietary Restriction, NTP TR 460 377 

I 

I 

;~ ~ .... i 

" I 

I 

I = 

:: : i 

.i ~ 

; I 

:;4 I : ~ 

:::5 [ 

I 

=I 1 

:: 1 

1 

;= I 

5:1 

:: I 

:; I 

1 

:: I 

TABLE I4b 

Organ Weights and Organ-Weight-to-~ody-Welght Ratios for l~Ia]e I~Iice. 

at the 15-1Vl~onth ~nter~m Evaluation in the Dietary ]Restrict~o~ Study of Sa~icy~a~osu~fapyr~d~ae: 

2-Year and 3-Year Reestr~cted Feed ~hrotoco~s a 

Vehicle Control 2,700 mg/kg 

n 10 10 

Necropsy body wt 47.7 5:1.7 34.4 5: 0.6** 

R. KidneY 

Absolute 0.325 5:0.013 0.279 5: 0.011" 

Relative 6.85 5:0.28 8.14 5: 0.38* 

Liver 

Absolute 1.758 5:0.097 1.757 5:0.034 

Relative 36.75 5:1.26 51.18 5: 1.10"* 

Spleen 

Absolute 0.073 =1:0.004 0.124 5: 0.007/** 

Relative 1.53 5:0.07 3.62 5: 0.23** 

R. Testis 

Absolute 0.122 5:0.004 0.112.5: 0.002* 

Relative 2.58 5:0.11 3.27 5: 0.10"* 

Thyroid gland 

Absolute 0.003:5:0.000 0.003 ::1:0.000 

Relative 0.06 5:0.01 0.08 5:0.01 

* Significantly different (Pg0.05) from the control group by Student's t-test 

** P~0.01 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean :1: standard error). 
378 Dietary Restriction, NTP TR 460 " 

TA2aLE 15a 

Organ Weights and Organ-Weight-t0-Body-Weight Ratios for Mice at the 15-Month Interim Evaluation 

in the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad ILibitum. Weight-Matched 

Fed Control Control 25 mg/kg 

Male 

9 10 10 

Necropsy body wt 50.2 ± 0.6 39.3 ± 1.3 39.0 ± 0.6** 

R. Epididymis 

Absolute 0.062 ± 0.003 0.057 ± 0.003 0.057 ± 0.003 

]Relative 1.23 ± 0.05 1.47 ± 0.08 1.45 ± 0.06* 

R, Kidney 

Absolute 0.376 ± 0.015 0.303 ± 0.010 0.325 ± 0.006** 

Relative 7.47 ± 0.22 7.73 ± 0.22 8~32 ± 0.09**, 

Liver 

Absolute 2.336 ± 0.122 1.300 ± 0.051 2.152 ± 0.353, 

Reladve 46.54 ± 2.34 33.09 ± 0.91 55.43 ± 9.41, 

R. Testis 

Absolute 0.119 ± 0.003 0.116 ± 0.004 0.117 ± 0.003 

Relative 2.37 ± 0.05 2.98 ± 0.16 2.99 ± 0.04** 

Female 

n 10 10 10 

Necropsy body wt 53.2 ± 1.9 41.4 ± 1.2 40.7 ± 1.7"* 

R. Kidney 

Absolute 0.256 ± 0.005 0.233 ± 0.004 0.237 ± 0.008 

Relative 4.83 ± 0.10 5.68 ± 0.21 5.87 ± 0.16"* 

Liver' 

Absolute 1.915 ± 0.058 1.613 ± 0.023 1.820 ± 0.051,A 

Relative 36.09 ± 0.58 39.27 ± 1.19 45.15 ± 1.50**A, 

* Significantly differem (P~0.05) from the ad libitum-fed control group by Student's t-test 

** P~0.01 

, Significantly different (P~0.05) from the weight-matched control group by Student's t-test 

,, P~0.01 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean ± standard error). 
Die~ry Restriction, NTP TR 4~0 37~ 

TABLE ISb 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice at the 15-Month ~nterhn Evaluation 

in the Dietary Restriction Study of Scopolamine Hydrobromide Tr~hydrate: . 

2-'Year and 3-Year ~es~r~cted Feed ]Protocols a 

• ~ 

Vehicle Control 25 mg/kg 

n 10 10 

Male 

Necropsy body wt 38.2 5:0.5 36.6 5: 0.6* 

R. Epididymis 

Absolute 0.055 5:0.002 0.053 5:0.003 

Relative 1.43 5:0.04 1.46 ::1:0.07 

R. Kidney 

Absolute 0.340 5:0.006 0.314 5: 0.004** 

Relative 8.91 5:0.18 8.60 5:0.09 

Liver 

Absolute 1.766 5:0.016 1.714 5:0.023 

Relative 46.25 5:0.52 46.94 5:0.81 

R. Testis 

Absolute 0.120 5:0.002 0.117 5:0.003 

Relative 3.13 5:0.06 3.19 5:0.06 

Female 

Necropsy body wt 35.8 5:0.8 34.4 5:0.6 

R. Kidney 

Absolute 0.238 5:0.003 0.243 5:0.007 

Relative 6.68 5:0.15 7.06 5:0.23 

Liver 

Absolute 1.630 5:0.031 1.619 5:0.025 

Relative 45.64 5:1.14 47.14 5:0.89 

* Significantly different (P~0.05) from the control group by Student's t-test 

** P~O.OI 

a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as 

mg organ weight/g body weight (mean 5: standard error). 
380 Dietary Restriction, NTP TR 460 

rr • 

J 

J~/ 

r 
38~. 

 ̧ : i 

:: ̧ I 

: I 

TABLE Jla 

TABLE Jib 

TABLE J2a, 

TABLE J2b 

TABLE J3a 

TABLE J3b 

TABLE J4a 

TABLE J4b 

TABLE JSa 

TABLE JSh 

A I EI ]IX 

I IEAN I ©I Y 

ANI ..f V IVAIL I dE U]LT 

Mean Body Weights and SurvivaH of Rats 

an the Dietary Restriction Study of Butyl BenzynPhthanate: 

Ad Libitum Feeding and Welght-Matched Controls Protocols ................... 382 

Mean Body Weights and Survival of Rats 

~n the Dietary Reestriction Study of Butya ]Benzy~ ]?hth~flate: 

2-Year and Long-Te~ R~tricted ~eed Protocols .......................... 384 

Me~ Body Weights and Su~val of Rats 

an the Dietary Restriction Study of t-Butylhydroqu~none: 

Ad Libimm Feeding and Weight-Matched Control Prot~o~ ................... 386 

Mean Body Weights and Surv~van of RaN 

m the Dietary ~triction Study of t-Butylhydroquinone: 

30-Month R~tr~cted ~eed Protocol ......... . .......................... 388 

Me~ ~ody Weights and Survival of Male ~ats 

~n the D~eta~ R~tr~ction Study of Salicylazosulfapyr~d~ne: 

Ad L&i~m ~eeding and Weight-Matched Controls ~ot~o~ ................... 3~0 

Mean Body Weights and Survival of Mane RaN 

m the Dietary Restriction Study of Salicy~azosulfapyr~d~ne: 

2-Year and 30-Month Restricted Feed Protocols ............................ 391 

Mean Body Weights and Survival of Male Mice 

m the Dietary R~triction Study of Salicylazosulfapyr~d~ne: 

Ad Libi~m Feeding and Weight-Matched Controls Protocol ................... 392 

Mean Body Weights and Survival of Male Mice 

m the Dietary Restriction Study of Salicylazosulfapyr~dine: 

2-Year and 3-Year Restricted Feed Protocols .............................. 393 

Mean Body Weights and Survival of Mice 

m the Dieta~ Restriction Study of Scopolamine Hydrobrom~de ~hydrate: 

Ad L~i~m Feeding and Weight-Matched Controls ~otoco~s ................... 395 

Mean Body Weights and Survival of Mice 

in the Dietary Restriction Study of Scopolamine Hydrobrom~de Tr~hydrate: 

2-Year and 3-Year Restricted Feed Protocols .............................. 39'7/ 
382 Dietary Restriction, NTP TR 460 

TABLE Jla 

iMean Body Weights and Survival of Rats in the Dietary Restriction Study 

of ]Butyl Benzyl Phthalate: Ad Libitum Feeding.and Weight-MatchedControls Protocols 

Ad L/b/turn-Fed Weight-Matched 

Weeks Control ~. (~ontrol 12.000 ~nm 

on Av. Wt. No. of Av. wt. No. of Av. Wt. Wt. (% of Wt. (% of No. of 

Study (g) Survivors (g) Survivors (g) ad//b/turn-fed weight-matched Survivors 

controls) controls) 

Male 

1 113 60 110 60 108 96 98 

2 149 60 146 60 141 95 97 

3 183 60 181 60 180 98 99 

4 '205 60 210 60 208 101 99 

5 233 60 234 60 226 97 97 

6 253 60 252 60 243 96 97 

7 266 60 255 96 100 

8 286 60 282 60 272 95 96 

9 298 60 297 .60 284 95 96 

10 310 60 306 60 291 94 95 

I1 319 60 319 ~60, 301 94 94 

12 326 60 325 60 312 96 96 

13 339 60 335 60 315 93 94 60 

17 370 60 365 60 338 91 93 60 

21 391 60 352 60 351 90 I00 60 

25 391 60 364 60 362 93 100 6O 

29 416 60 381 60 377 91 99 59 

33 423 60 399 60 382 90 96 59 

37 429 60 395 60 389 91 98 59 

41 437 58 378 60 394 90 104 59 

45 443 58 391 60 405 92 104 58 

49 449 58 365 60 413 92 113 58 

53 451 58 400 59 414 92 104 58 

57 450 58 391 59 412 92 105 58 

61 45l 58 413 59 413 92 100 58 

65 451 57 412 58 420 93 102 58 

69 a 446 46 422 47 419 94 99 46 

'7'3 448 45 427 45 427 95 100 45. 

'7'7 446 45 427 45 428 96 100 45 

81 440 42 432 45 422 96 98 44 

85 440 40 433 44 418 95 97 44 

89 439 36 438 43 413 94 94 42 

93 440 33 438 42 422 96 96 39 

97 433 32 434 38. 411 95 95 35 

101 426 31 433 36 402 94 93 29 

Mean for weeks 

1-13 252 250 241 96 96 

14-52 417 • 377 379 91 101 

53-101 443 423 417 94 99 
Die~-y ~es~r~cfion~ ~ T~ ~6~ 383 

TA~L~ ,lIla 

I~,~Ie~n ~ody Weights and Surviva~ of Rats ~u the Dietary Restriction Study 

of ~uty~ ~enzy~ Phth~flate: Ad Libitum Feeding and Weight-I~,lIatched Controls ]Protocols (continued) 

Ad Libitum-Fed Weight-Matched 

Weeks Control Control 2~.000 ~m 

on Av. Wt. No. of Av. Wt. No. of Av. Wt. Wt. (% of Wt. (% of No. of 

Study (8) ~urvivors (g) Survivors (g) ad//b/tum-fed weight-matched Survivors 

controls) controls) 

Female 

1 99 60 97 60 92 93 95 60 

2 ' 121 60 120 60 114 94 95 60 

3 135 60 134 60 127 94 95 60 

4 142 60 142 60 137 97 96 60 

5 153 60 151 60 142 93 94 60 

6 159 59 159 60 151 95 95 60 

7 167 59 166 60 157 94 95 60 

8 171 59 170 60 160 94 95 60 

9 175 59 174 60 166 95 96 60 

10 178 59 180 60 171 96 95 60 

11 180 59 183 60 170 95 93 60 

12 186 59 186 60 175 94 94 60 

13 185 59 184 60 176 95 96 60 

17 198 59 193 60 183 93 95 60 

21 204 59 203 60 187 92 92 60 

25 213 59 209 60 193 91 92 60 

29 218 59 221 60 197 91 89 60 

33 223 59 211 59 199 89 94 60 

37 231 59 204 59 203 88 99 60 

41 238 59 198 59 206 87 104 60 

45 240 59 196 59 207 86 106 60 

49 258 59 194 59 212 82 109 60 

53 269 59 199 59 214 80 108 60 

57 276 58 200 59 214 78 107 60 

61 281 58 202 59 216 77 107 59 

65 286 58 203 59 220 77 109 59 

69 a 296 48 216 49 222 75 103 46 

73 298 48 208 48 226 76 109 44 

77 300 47 228 48 230 77 101 44 

81 300 47 228 47 222 74 97 43 

85 312 45 239 46 231 74 97 43 

89 321 44 245 46 237 74 97 39 

93 320 44 238 46 236 74 99 39 

97 319 42 247 46 233 73 94 39 

I01 318 36 254 46 232 73 922 32 

105 236 42 247 105 29 

Mean for weeks 

I 1-13 158 157 149 94 95 

I 14-52 225 203 199 88 98 

53-105 300 225 227 76 lOl 

a Interim evaluation occurred during week 66. 
38,1 Dietary Restriction, NTP TR 460 

• 

• .-. TABLE Jlb • :

Mean Body Weights and Survival of Rats .. ~') in the Dietary Restriction Study • of Butyl Benzyl Phthalate: 2-Year and, Long-Term Restricted Feed Protocols -. :

2-Year Protocol 30-Month Prot0cgl " 

Weeks 0 uum 12s000 ppm 0 ppm 12.000 uDm 

• Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. No. of Av. Wt. Wt. (% of No. of 

Study (g) Survivors (g) controls) Survivors (g) Survivors (g) cont~0ls) Survivors 

on 

~, 

Ma~ 

1 103 60 101 98 60 103 50 101 98 50 

2 140 60 138 99 60 139 50 138 99 

3 146 60 143 98 60 146 50 143 98 5O 

4 156 60 155 99 60 155 50 154 100 5O 

5 176 60 168 95 60 182 50 167 92 5O 

6. 200 60 180 90 60 204 50 179 89 5O 

7 208 60 199 96 60 210 •50 204 97 5O 

8 227 60 216 95 60 233 50 217 94 5O 

9 " 246 60 234 95 60 246 50 244 99 49 

10 260 60 248 95 60 262 50 254 97 49 

11 258 60 249 97 60 261 50 258 99 ~.9. 

.12 261 60 251 96 60 263 50 256 97 49 

13 285 60 274 96 60 287 50 280 97 49 

17 301 60 294 98 60 307 50 303 99 4~ 

•21 , 339 60 324 96 60 349 50 323 93 .48 

25 358 60 339 95 60 363 50 342 94 48 

29 349 50 340 98 60 356 50 344 97 48 . 

33 362 60 347 96 60 376 50 351 93 48 " 

37 366 60 344 94 60 378 50 346 92 48 

41 383 60 349 91 60 390 50 351 90 48 

45 ~ 372 60 345 93 60 376 50 357 95 48 

49 366 60 343 94 60 371 50 344 93 48, 

53 368 60 351 95 59 376 50 357 95 48 

57 378 60 370 98 59 398 50 373 ,94 48~ 

61 399 .60 375 94 59 404 50 378 94 48 

65 388 59 374 96 59 405 50 379 94 48 

69 a 406 48 377 93 49 408 50 381 94 48 

73 403 48 378 94 49 410 50 374 91 

77 377 47 354 94 48 376 49 347 92 44 

' 8f 395 46 372 94 48 389 49 364 9~ 4~: 

8~ 398 45 362 91 48 399 48 364 91 41 

89 371 44 342 92 45 371 46 345 93 40' 

93 365 42 362 99 41 378 44 367 97 37 

97 371 41 341 92 39 386 40 348 90 36 

101 354 38 340 96 35 370 37 342 93 33 

105 360 32 334 93 30,. 

109 359 26 327 91 26 

11~ 359 20 332 93 24 

117 332 16 314 94 19 

121 324 12 304 94 18 

125 329 11 307 93 14 

Mean for weeks -, 

1-13 205 197 96 200 97 

340 94 . ,

53-101 383 361 94 390 363 93 

102~125 344 320 93 

14-52 355 336 95 363 
Dile~ry ~estrfiction, NTP TR 460 385 

T~L~ dish 

Mee~u Body Weights and Survival of ]Rats ~n the Dietary ]Restriction Study 

off' ~utyH ~enzy~ Phth~ate: 2-Year and Long-Term Restricted Feed IProtoco~s (continued) 

2-Year Protocol 32-Monl~h 1protocd 

Weeks 0 ham 2~4.000 yam 0 m~m 24.~ a.am 

on AV. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. No. of Av. Wt. Wt.(%of No. of 

I Study (g) Survivors (g) controls) Survivors (g) Survivors (g) controls) Survivors 

I : ~ I ~ I 

I Female i 

I 1 94 60 88 93 60 95 50 87 91 50 

2 .116 60 107 92 60 118 50 105 89 50 

3 128 60 118 92 60 128 50 116 91 50 

,4 131 60 124 95 60 131 50 122 93 50 

~ I 5 133 60 127 96 60 134 50 127 95 50 

i , I i~: ~ ] 6 146 60 134 92 60 146 50 133 91 50 

~ ~ I 7 141 60 130 92 60 142 50 130 91 50 : I 

i 

8 141 60 141 100 60 141 50 139 98 50 

9 159 60 148 93 60 158 50 147 93 50 

: ! 10 155 60 145 94 60 155 50 145 93 50 

il 157 60 146 93 60 157 50 146 93 50 

12 160 60 152 95 60 161 50 151 94 50 

!3 162 60 155 96 60 163 50 151 92 50 

17 170 60 164 97 60 170 50 16~ 96 50 

21 176 60 164 93 60 180 50 164 91 50 

: I 25 189 60 178 94 60 188 50 176 93 50 z ~ I 

29 193 60 180 93 60 194 50 180 93 50 ~!; :: I 

33 194 60 175 90 60 195 50 173 89 50 

37 182 60 176 97 60 183 50 180 99 50 

41 191 60 175 92 60 199 50 173 87 50 

::1 45 190 60 180 95 60 192 50 178 93 50 

49 198 59 184 93 60 200 50 184 92 50 

53 206 58 191 93 60 209 49 191 91 50 

57 217 58 194 90 60 221 49 191 87 50 

61 222 58 198 89 60 227 49 196 86 50 

65 228 58 197 87 59 232 49 197 85 50 

!::l 69 a 233 48 202 87 49 238 48 199 84 50 

: 73' 244 48 208 85 48 247 46 203 82 50 

77 254 48 208 82 48 257 46 207 81 49 i!i 81 263 46 210 80 48 269 46 205 76 49 

12 85 273 46 211 77 48 280 45 206 73 49 

2 : I 89 276 44 212 77 46 284 45 207 73 49 

I 93 2~2 42 212 75 46 2~9 45 210 73 47 

: ! 97 284 41 211 74 43 294 43 210 72 47 

101 276 37 213 77 41 293 37 209 71 44 

105 288 34 210 73 39 

109 290 28 211 73 36 

113 283 28 211 75 34 

i17 283 26 210 74 31 

i21 282 23 208 74 27 

125 291 19 206 71 22 I 

?: i ;;22 ] Mean for weeks 

1-13 140 132 94 141 131 93 

14-52. 187 175 94 189 175 93 

53-101 251 205 82 257 202 79 

102-125 286 209 73 

a Interim evaluation occurred duringweek 66 of the 2-year protocol. 
386 Dietary Restriction, NTP TR 460 

T~BLE J2a 

Mean Body Weights and Survival of Rats in the Dietary Restriction Study oft-Butylhydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols 

Weeks 

on 

Study 

Ad L/b/tum-Fed 

Control 

Av. Wt. No. of 

(g) Survivors 

Weight-Matched 

Con|r9| 

Av. Wt. No. of 

(g) Survivors 

Av. Wt. 

(g) 

5.000 m~m 

Wt. (% of Wt. (% of 

ad//b/tum-fed weight-matched 

controls) controls) 

No. of 

Survivors 

Male 

1 99 70 97 70 91 92 94 70 

2 141 70 139 70 127 90 91 70 

3 175 70 161 70 157 90 98 70 

4 209 70 188 70 188 90 100 70 

5 233 70 189 70 217 93 115 70 

6 267 70 215 70 244 91 113 70 

7 283 70 247 70 261 92 106 70 

8 299 70 250 70 275 92 110 70 

9 313 70 260 70 287 92 110 70 

10 316 70 273 70 299 95 109 70 

11 324 70 296 70 304 94 103 70 

12 344 70 314 70 318 92 101 70 

13 356 70 323 70 320 90 99 70 

17 a 379 60 364 60 346 91 95 60 

21 400 60 336 60 358 89 106 60 

25 413 60 348 60 376 91 108 60 

29 419 60 384 60 386 92 100 60 

33 428 60 380 60 392 92 103 60 

37 437 60 389 60 402 92 103 60 

41 450 60 393 60 416 92 106 60 

45 452 60 406 60 415 92 102 60 

49 444 60 398 60 415 94 104 60 

53 468 60 417 60 431 92 103 60 

57 468 59 415 59 435 93 105 60 

61 467 59 419 59 440 94 105 60 

65 470 59 429 59 440 94 103 59 

69 468 59 428 59 437 93 102 59 

73 472 56 423 59 433 92 102 57 , 

77 463 55 435 59 438 95 101 55 

81 464 52 432 59 436 94 101 53 

• 85 462 51 428 58 429 93 100 52 

89 455 51 431 54 425 93 99 51 

93 458 46 429 51 419 92 98 44 

97 455 39 434 45 421 92 97 39 

101 447 35 430 41 421 94 98 34 

105 440 28 426 35 409 93 96 30 

109 420 21 416 30 412 98 99 25 

113, 429 13 419 27 395 92 95 19 

117 428 10 399 23 397 93 99 15 

121 417 8 406 14 386 93 95 14 

Mean for weeks 

1-13 258 227 238 92 105 

14-52 425 378 390 92 103 

53-121 453 423 422 93 100 
Dietary ]Restr~cfion, NTP TR 46~ 387 

'I~AnLE g2a 

Mean Body Weights and Survival of Rats ~n the Dietary Restriction Study of t-~uty~hydroqe.finone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad L/b/turn-Fed Weight-Matched 

Weeks Control Control ~.~0 n~a~ 

on Av. Wt. No. of Av. Wt. No. of Av. Wt. Wt. (% of Wt. (% of ~0. Of 

: ̧ i Study (g) Survivors (g) Survivors (g) ad//b/turn-fed weight-nmtched Survivors 

~ 1 controls) controls) 

~ ] Female ~ ] 

; ] 

1 92 70 92 70 83 91 91 70 

iii~ I~ 2 121 70 121 70 110 91 91 70

ili !i! ] 3 135 70 126 70 125 92 99 70 

::~iiiii I 4 147 70 130 70 134 92 104 70 

~!!~ ~i ~ I 5 159 70 123 70 146 92 119 70 

I 6 171 70 129 70 154 90 120 70 

[ 7 177 70 155 70 161 91 104 70 

....I 8 181 70 161 70 165 92 103 70 

!:i!~] 9 186 70 149 70 173 93 116 70 iii il: 10 176' 70 150 70 176 100 118 70 

::~ i 11 193 70 168 70 181 94 108 70 :,z ]

• 12 196 70 175 70 181 92 103 70 

: ~i~ii I 13 198 7o 180 70 184 93 1o2 70 

~ I 17 a 203 60 "192 , 60 188 93 98 60 

~i] I 21 224 60 192 60 200 89 104 60 

i)! 25 222 60 207 60 203 91 98 60 

29 229 60 213 60 210 91 98 60 

~:11 33 231 60 210 60 210 91 100 60 

iiii!~i] 37 232 60 219 60 213 92 97 60 

i?!i 41 240 60 227 60 219 92 97 60 

45 248 58 225 60 227 91 101 60 ,

!~i~ ] 49 257 58 231 60 232 90 100 60 

!!~ i~i ] 53 264 58 227 59 236 90 104. 60 

~:~::~i: ~I 57 279 57 236 59 246 88 104 60 

ii~ i I 61 285 55 240 58 257 90 107 60 

~:~[ 65 292 55 243 58 263 90 108 60 

' i 69 301 55 245 58 265 88 108 60 

i~: ~ 73 309 54 250 58 270 87 108 59 

77 315 54 259 56 274 87 106 59 ! 

ii~1 81 323 51 257 56 278 86 108 59 

I 85 327 49 266 56 282 87 106 57 

89 333 48 272 54 287 86 106 54 

f~ 93 342 43 274 51 290 85 106 50 

97 348 42 270 50 299 86 111 46 

~1 101 347 39 282 50 301 87 107 43 ::~i i ~i~ I 

i± ~! 105 345 35 285 46 303 88 106 42 

~!i] 109 341 31 287 42 300 88 104 40 

] 113 337 24 295 39 301 89 102 37 

:!~i!!1 117 348 18 295 35 309 89 105 33 

121 341 16 294 31 315 92 107 27 

Mean for weeks 

1-13 164 143 152 93 106 

14-52 232 213 211 91 99 

53-121 321 265 282 88 106 

a Interim evaluation occurred during week 14. 
388 Dietary Restriction, NTP TR 460 

TABLE J2b 

Mean Body Weights and Survival of Rats in the Dietary Restriction Study 

of t-Butylhydroquinone: 30-Month Restricted Feed Protocol 

Weeks 0 ham 5,000 uam 

on Av. Wt. Number of Av. Wt. Wt. (% of Number of 

Study (g) Survivors (g) controls) Survivors 

Male 

1 

2 

3 

4 

5 

6 

7 

8 

9 

I0 

11 

12 

13 

17 a 

21 

25 

29 

33 

37 

41 

45 

49 

53 

57 

61 

65 

69 

73 

77 

81 

85 

89 

93 

97 

101 

105 

109 

113 

117 

121 

125 

129 

Mean for weeks 

1-13 

14-52 

53-129 

107 

151 

185 

185 

195 

208 

231 

246 

255 

266 

274 

289 

291 

314 

351 

343 

348 

358 

371 

390 

404 

403 

386 

392 

395 

398 

399 

395 

382 

393 

395 

401 

400 

403 

401 

369 

383 

370 

364 

360 

350 

324 

222 

365 

383 

70 

70 

70 

70 

70 

70 

70 

• 70 

70 

70 

70 

70 

70 

6O 

60 

6O 

60 

60 

60 

0 

6O 

60 

6O 

60 

60 

60 

60 

59 

59 

58 

55 

51 

46 

43 

39 

35 

30 

26 

18 

13 

13 

1.1 

94 

139 

167 

163 

179 

195 

216 

229 

240 

250 

260 

276 

281 

309 

341 

342 • 

344 

" 362 

370 

387 

397 

393 

391 

394 

391 

396 

395 

389 

378 

393 

39O 

397 

391 

389 

387 

364 

372 

367 

• 348 

348 

343 

315 

207 

361 

~377 

88 

92 

90 

89 

92 

94 

94 

93 

94 

94 

95 

96 

97 

98 

97 

100 

99 

101 

100 

99 

99 

98 

I01 

100 

99 

100 

99 

99 

99 

100 

99 

99 

98 

97 

97 

99 

97 

99 

96 

97 

98 

97 

93 

99 

98. 

70 

70 

70 

70 

70 

70 

70 

70 

70 

70 

70 

70 

70 

6O 

6O 

6O 

60 

60 

0 

6O 

60 

60 

60 

6O 

6O 

6O 

60 

6O 

6O 

60 

6O 

59 

57 

56 

53 

50 

41 

39 

34 

29 

26 

24 
I Dietary Restriction, ~ TR 46~ 389 

i I 

I TABLE J2b 

Mean ~ody Welgh~s and Smwiva~ of Rats in the Dietary ]Restriction Study 

of t-~uty~hydroquinone: 30-Mouth Restricted Feed ~otoco~ (continued) 

i 

i 

Weeks 0 yam 5.000 vvm 

on Av. Wt. Number of Av. Wt. Wt. (% of Number of 

Study (g) Survivors (g) controls) Survivors 

~ I ~ ! 

~:l 

Female :: ! 

~ I 1 92 70 82 89 70 

I 2 128 70 120 94 70 ; 1 

3 129 70 120 93 70 ~? i 

4 128 70 120 94 70 

5 132 70 124 94 70 

6 134 70 129 97 70 

7 141 70 135 96 70 " 1 

8 146 70 141 96 70 

= 

5 i ! 

~ 1 9 150 70 145 96 70 = ;ii I 

~ :1 10 153 70 144 94 70 1 

I 11 157 70 148 94 70 

12 162 70 155 96 70 

13 181 70 175 97 70 

17 a 172 60 171 I00 60 

21 189 60 188 99 60 

~! 25 181 60 183 101 60 

29 184 60 183 99 60 c I 33 194 60 195 101 60 ; ::; ! 

37 212 60 213 100 60 = :! 

41 213 60 214 100 60 

I 45 208 60 210 101 60 

2: I 49 209 60 211 101 60 ~::-- I 

53 204 60 204 100 60 

~ ] 57 221 60 224 101 60 

~ ] ~ , 61 224 60 226 101 59 ~ ] 65 231 60 235 102 59 ! 69 236 60 237 101 59 :? I 

; i 73 240 59 243 101 58 c~ 1 : ~ 77 244 59 244 100 58 

1 81 254 58 258 102 57 ~ i 

~ ~ 85 264 57 262 100 57

: i 89 279 56 274 98 55 

ii;l 93 294 53 283 96 55 

:; i :i 97 302 52 290 96 52 ~75 ] 101 308 49 296 96 50 :~i~l 105 317 44 289 91 49 

109 315 43 295 94 47 1 113 317 38 296 94 42 : ) 117 308 35 285 93 40 

= 1 121 300 27 278 93 35 

125 285 20 273 96 27 : , I 

.... I : ; ' 129 270 18 264 98 25 

~ I 

Mean for weeks 

1-13 141 134 95 

14-52 196 196 100 

53-129 271 263 97 

Interim evaluation ¢wcurred during Week 14. 

,~:) i 
390 Dietary Restriction, NTP TR 460 

T~SL~ J3a 

Mean Body Weights and Survival of Male Rats in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols ~ ~ 

Ad L/b/turn-Fed Weight-Matched 

Wt~ks Control • Control 337.5 n~/kt, 

on Av. Wt. No. ef Av. Wt. No. of Av. Wt. Wt. (% of Wt. (% of No. of 

Study (g) Survivors (g) Survivors (g) ad//b/iron-fed weight-matched Survivors 

controls) controls) 

1 102 70 103 60 102 100 99 60 

2 139 70 136 60 135 97 100 60 

3 171 70 174 60 173 101 99 60 

4 196 70 200 60 195 100 98 60 

5 216 70 220 60 215 100 98 60 

6 238 70 243 60 240 101 99 60 

7 258 70 259 60 259 100 100 60 

8 273 70 273 60 275 101 101 60 

9 287 70 286 60 280 97 98 60 

10 299 70 298 60 296 99 99 60 

11 307 70 305 59 301 98 99 60 

12 317 70 316 59 313 99 99 60 

13 324 70 324 59 316 97 97 60 

14 333 70 337 59 331 100 98 60 

18 360 70 364 59 358 99 98 60 

21 374 70 371 59 363 97 98 59 

25 393 69 394 59 386 98 98 59 

29 a 416 59 410 59 400 96 98 59 

33 424 59 425 59 413 97 97 59 

37 439 59 431 59 423 96 98 58 

41 447 59 437 59 428 96 98 58 

45 457 59 451 59 439 96 97 58 

49 461 59 459 59 444 96 97 58 

53 469 59 463 59 451 96 98 58 

57 471 59 467 59 460 98 99 58 

61 477 59 468 59 453 95 - 97 58 

65 477 59 474 59 459 96 97 56 

69 a 474 48 471 48 461 97 98 45 

73 477 46 470 47 463 97 98 45 

77 472 45 469 45 451 96 96 45 

81 468 44 468 44. 44.9 96 96 45 

85 467 44 458 43 450 96 98 42 

89 461 43 460 42 444 96 97 41 

93 453 42 449 40 439 97 98 39 

97 442 41 437 39 437 99 100 34 

101 422 39 426 35 417 99 98 31 

Mean for weeks 

1-13 241 241 238 99 99 

14-52 410 408 399 97 98 

53-101 464 460 449 97 98 

a Interim evaluation occurred during weeks 27 (ad libitum-fed controls only) and 66. 
 ̧= i 

Dietary Restriction, N'YP TR ~ 39~. 

T~L~ $3b 

Mee~ ~ody Weighs ~nd Survived o~ I~JI~e ~,~ ~n the Die~.m'y ll~es~taSct~on S~u~y 

o~' ~icy~osu~'~pyr~ne: 2-Ye~" zn~ 30-I~on~h ]~estr~cte~ ~eed~ ~Ihrotcco~s 

I 2-Year Protocol 30-1Month Protofgl 

Weeks Vehicle C~ntr01 337.5 m~__~.~ Vehicle Control 337.5 nuff_~ 

..... ~ 

: l

]

! 

on 

Study 

Av. Wt. 

(g) 

No. of 

Survivors 

Av. Wt. 

(g) 

Wt. (% of No. of 

controls) Survivors 

Av. Wt. 

(g) 

No. of 

Survivors 

Av. Wt. 

(g) 

Wt. (% of I~Io. of 

controls) Survivors 

~ i 

I 107 61 108 102 60 109 49 109 100 50 

2 140 61 140 1130 60 143 49 141 99 50 

3 167 61 167 1130 60 171 49 168 98 50 

4 183 61 181 99 60 183 49 181 99 50 

5 . 205 61 205 100 60 204 49 204 100 50 

6 216 61 215 99 60 217 48 217 100 50 

7 230 61 230 100 60 232 48 231 100 50 

8 241 61 240 99 60 243 48 238 98 50 I 9 250 61 248 99 60 253 48 245 97/ 50 

10 261 61 255 98 60 261 48 254 97 50 

11 270 61 267 99 60 273 48 264 97 50 

I 12 283 61 281 99 60 289 48 277 96 50 

i 14 

17 

298 

306 

61 

61 

296 

298 

99 

97 

59 

59 

302 

315 

48 

48 

290 

295 

96 

94 

50 

50 

20 318 61 308 97 59 320 48 306 95 49 

24 336 61 324 96 59 339 48 319 94 49 

28 345 61 325 94 58 344 48 324 94 49 :r I 32 359 61 341 95 57 357 48 339 95 47 

37 363 61 343 95 57 363 48 347 96 47 

40 370 61 354 96 57 375 48 351 94 47 

I 44 383 61 356 93 57 381 48 357 94 47 

I 48 384 61 359 94 57 387 47 362 93 47 

53 383 61 360 94 57 384 47 361 94 47 

56 408 61 382 94 57 408 47 385 94 47 

60 420 61 387 92 57 422 47 388 92 47 

2 ;

: I ! 64 425 60 382 90 57 424 46 381 90 46 

:~ 

I I 68 a 

72 

427 

423 

50 

49 

386 

378 

90 

89 

47 

47 

433 

425 

45 

45 

386 

378 

89 

89 

46 

46 : 

:? I I 76 409 48 361 88 45 408 45 361 88 46 

i ! 80 403 48 349 87 45 4O7 44 348 86 46 

; ; 

:: I I 

I 

84 

89 

92 

405 

402 

396 

48 

47 

44 

345 

342 

340 

85 

85 

86 

45 

43 

42 

401 

400 

394 

43 

42 

41 

348 

346 

341 

87 

87 

87 

43 

43 

42 =~ 96 400 41 339 85 42 398 39 340 85 42 

100 400 36 329 82 40 386 38 329 85 41 

104 384 34 317 83 40 388 35 327 84 41 

108 375 33 311 83 39 

112 380 29 304 80 39 

116 365 25 295 81 35 

120 351 19 282 80 32 

124 335 18 273 81 29 

128 304 13 255 84 24 

Mean for weeks 

1-13 213 211 99 215 211 98 

14-52 346 330 95 348 329 95 

53-104 406 357 88 406 359 88 

105-128 352 287 82 

a Interim evaluation occurred during week 66 of the 2-year protocol. 
392 Dietary Restriction, NTP TR 460 

T~BLE J4a 

Mean Body Weights and Survival of Male Mice in the Dietary Restriction Study 

of Salicylazosulfapyridine: Ad Libitum Feeding and Weight-Matched Controls Protocols 

Ad L/b/turn-Fed Weight-Matched 

Weeks Control C~ntrol 2.700 nm/k~ 

tm Av. Wt. No. of Av. Wt. No. of Av. Wt. Wt. (% of Wt. (% of No. of 

Study (g) Survivors (g) Survivors (g) ad//b/turn-fed weight-matched Survivorscontrols) controls) 

1 23.5 

2 25.3 

3 25.9 

4 27.7 

5 28.4 

6 29.1 

7 30.0 

8 31,4 

9 31.6 

I0 32.0 

11 32.9 

12 33.5 

13 34.6 

14 35.2 

18 38.2 

22 40.7 

26 44.7 

30 45.2 

34 47.7 

38 48.7 

42 49.2 

46 49.7 

50 50.2 

54 51.7 

58 52.6 

62 52.8 

66 a 53.3 

68 a 

70 54.0 

74 53.6 

78 54.4 

82 54.3 

86 52.7 

90 52.9 

94 53.4 

98 52.3 

102 52.0 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

59 

59 

59 

59 

59 

59 

59 

58 

58 

56 

46 

46 

46 

46 

46 

46 

44 

44 

41 

40 

23.4 

24.7 

26.0 

27.3 

28.0 

28.5 

29.0 

30.2 

31.0 

31.0 

31.9 

32.3 

33.7 

33.5 

36.5 

40.1 

42.4 

37.5 

35.3 

39.6 

41.7 . 

45.1 

42.2 

43.6 

45.3 

47.7 

47.1 

46.8 

45.1 

45.3 

43.8 

45.4 

49.0 

49.5 

49.9 

50.5 

Mean for weeks 

1-13 29.7 

14-52 45.0 

53-102 53.1 

29.0 

39.4 

46.8 

60 23.2 99 99 60 

60 25.0 99 101 58 

60 26.6 103 102 58 

60 27.7 100 102 58 

60 28.1 99 100 58 

60 28.3 97 99 58 

60 29.5 98 102 58 

60 30.4 97 101 58 

60 30.7 97 99 58 

60 30.8 96 99 58 

60 31.7 96 99 58 

60 31.4 94 97 58 

60 32.5 94 96 58 

60 32.5 92 97 58 

60 34.2 90 94 58 

60 37.0 91 92 .58 

59 37.9 85 89 58 

59 39.2 87 105 58 

59 39.4 83 112 58 

59 40.0 82 101 58. 

59 40.1 82 96 58 

59 41.0 83 91 58 

59 41.8 83 99 58 

59 43.1 83 99 58 

59 44.7 85 99 58 

59 45.9 87 96 58 

46.2 87 48 

49 

49 45.2 84 97 48 

49 48.2 90 107 47 

49 45.8 84 101 47 

49 47.2 87 108 47 

48 . 46.2 88 102 47 

46 46.8 89 96 47 

45 47.2 88 95 46 

45 44.8 86 90 46 

45 45.7 88 91 46 

28.9 97 100 

38.3 85 97 

45.9 86 98 

a Interim evaluation occurred during week 66. 
Dietary Restriction, .NTP TR 460 393 

I, 

TABLE J4b 

Mean Body Weights and Survival of Male IV~Jce in the Dietary Restriction Study 

of Sa~icylazosulfapyrldine: 2-Year and 3-Year ~estrlcted Feed Protocols 

2-Year Protocol 3-Year Protocol 

Weeks .Vehicle Control 2,700 m~kg Vehicle Control 2,700 m~J_k~ 

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. No. of Av. Wt. Wt. (% of No. of 

Study (g) Survivors (g) controls) Survivors (g) Survivors (g) controls) Survivors 

] 

1 23.4 62 23.2 99 60 23.7 48 23.5 99 50 

=( I 

2 

3 

25.1 

24.9 

62 

62 

24.7 

24.3 

98 

98 

60 

60 

25.3 

24.9 

48 

48 

25.3 

24.7 

100 

99 

50 

50 

4 23.4 62 22.8 97 60 23.3 48 23.1 99 50 

5 24.9 62 23.5 94 60 24.7 48 23.9 97 50 

I 

6 

7 

8 

9 

10 

11 

12 

13 

17 

18 

24.9 

25.9 

27.0 

27.1 

27.9 

27.8 

29.0 

29.4 

32.4 

32.9 

62 

62 

62 

62 

62 

62 

62 

62 

62 

62 

23.2 

23.7 

25.2 

25.5 

26.1 

26.5 

26.2 

27.2 

28.3 

28.6 

93 

92 

93 

94 

94 

95 

90 

93 

87 

87 

60 

60 

60 

60 

60 

60 

60 

60 

60 

60 

24.7 

25.6 

27.0 

26.8 

27.9 

27.7 

29.0 

29.3 

32.0 

32.4 

48 

48 

48 

48 

48 

48 

48 

48 

48 

48 

23.4 

23.9 

24.9 

25.8 

26.5 

26.7 

26.4 

27.4 

28.5 

28.7 

95 

93 

92 

96 

95 

96 

91 

94 

89 

89 

50 

50 

50 

49 

49 

49 

49 

49 

49 

49 

22 35.0 62 30.1 86 60 34.4 48 30.3 88 49 

26 35.8 62 31.4 88 60 35.3 48 31.6 90 49 

30 40.7 62 32.6 80 60 39.9 48 33.1 83 49 

34 44.7 62 34.5 77 59 43.5 48 34.8 80 49 

I 

38 

42 

46.0 

42.5 

62 

62 

33.8 

33.8 

74 

80 

59 

59 

45.1 

41.7 

48 

48 

33.6 

33.8 

75 

81 

49 

49 

46 41.1 62 34. I 83 59 40.4 48 34.4 85 48 

50 40.4 62 32.7 81 59 39.7 48 32.7 82 48 

54 42.6 62 33.8 79 59 42.0 48 33.6 80 48 

58 49.6 62 36.6 74 59 48.9 48 36.8 75 48 

62 49.5 62 35.8 72 59 48.5 48 36.0 74 48 

66 a 48.9 53 34.9 71 49 47.5 47 34.7 73 48 

70 47.8 52 34.6 72 49 46.4 47 34.5 74 48 

74 46.9 51 34.5 74 48 45.5 47 34.2 75 48 

78 45.0 51 33.6 75 48 43.9 47 33.6 77 48 

: ~ 82 43.9 51 32.2 73 48 42.9 45 32.1 75 48 ~ ? 86 41.8 51 31.0 74 48 40.4 44 31.2 77 48 

90 44.1 49 34.7 79 47 42.5 44 34.7 82 47 

94 45.8 47 34.8 76 44 44.3 44 34.4 ~78 46 

98 47.6 45 34.8 73 44 45.2 44 34.6 77 46 

102 49.4 45 35.8 73 44 46.5 44 35.2 76 46 

106 46.7 42 35.0 75 44 

110 46.5 39 35.6 77 44 

114 46.0 39 35.2 77 44 

118 45.3 35 33.6 74 ~44 

122 43.8 35 33.4 76 43 

126 43.0 30 34.0 79 43 

130 41.3 29 33.2 80 42 

134 41.4 27 33.6 81 4.0 

138 39.3 24 32.5 8~ 40 

142 39.3 23 32.8 84 40 

146 38.5 22 32.8 85 40 

150 36.2 22 30.5 84 38 

154 35.7 21 31.8 89 36 

Mean for weeks 

1-13 26.2 24.8 95 26.1 25.0 96 

14-52 39.2 32.0 82 38.4 32.2 84 

53-102 46.4 34.4 74 45.0 34.3 76 

103-154 41.8 33.4 80 

a Interim evaluation occurred during week 66 of the 2-year protocol. 
394 Dietary Restriction, NTP TR 460 

TABLE J5a 

Mean Body Weights and Survival of Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: Ad Libitum Feeding and Weight-Matched Controls Protocols 

Ad Libitum-Fed Weight-Matched 

Weeks Control Control 25 me/k~ 

on Av. Wt. No. of Av. Wt. No. of Av. Wt. Wt. (% of Vet. (% of No. of 

Study (g) Survivors (g) Survivors (g) ad//b/turn-fed weight-matched Survivors 

controls) controls) 

Male 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

17 

21 

25 

29 

33 

37 

41 

45 

49 

53 

57 

61 

65 

69 a 

73 

77 

81 

85 

89 

93 

97 

101 

24.3 

26.1 

26.6 

28.0 

29.2 

30.2 

30.9 

31.4 

31.9 

32.5 

33.4 

33.7 

34.5 

38.0 

40.9 

42.1 

43.8 

45.5 

47.9 

47.8 

49.7 

49.7 

49.7 

49.8 

50.5 

50.9 

51.7 

52.0 

52.4 

51.9 

51.4 

51.8 

51.2 

49.8 

49.7 

Mean forweeks 

1-13 30.2 

1,4-52 45.0 

53-101 51.0 

70 24.1 60 24.1 99 100 70 

70 23.4 59 25.4 97 109 70 

70 "22.7 56 25.7 97 113 70 

70- 25.8 54 27.0 96 105 70 

70 26.6 54 27.8 95 105 70 

70 27.5 54 27.8 92 101 70 

70 27.6 54 28.2 91 102 70 

70 28.9 54 28.5 91 99 70 

70 26.7 54 28.7 90 108 70 

70 26.4 54 29.2 90 111 70 

70 27.6 54 30.1 90 109 70 

70 27.4 54 30.5 91 111 70 

70 27.0 54 30.5 88 113 70 

70 28.5 54 31.9 84 112 70 

70 29.4 54 33.4 82 114 70 

70 34.8 54 34.0 81 98 70 

70 37.8 54 34.9 80 92 70 

70 34.8 54 35.6 78 102 70 

70 36.3 54 37.6 79 104 70 

70 39.3 54 38.3 80 98 70 

69 40.4 54 38.9 78 96 69 

69 42.1 54 39.7 80 94 69 

69 42.4 54 39.8 80 94 69 

69 43.5 54 39.1 79 90 69 

69 40.9 54 39.8 79 97 67 

67 39.5 54 40.8 80 103 66 

48 44.4 44 41.6 81 94 46 

48 42.9 44 40.8 79 95 46 

48 45.7 43 41.4 79 91 46 

47 45.7 43 41.1 79 90 46 

47 43.1 43 41.3 80 96 45 

47 40.8 42 41.3 80 101 44 

46 39.5 42 41.5 81 105 42 

46 39.3 42 41.0 82 104 42 

42 36.4 42 40.1 81 110 40 

26.3 28.0 93 106 

35.9 36.0 80 100 

41.9 40.7 80 97 
~ _ ~ ....... ~ ~ :~ ....... ~ , , ~ - - ~ ..... ~.---~-z~-~7, . ~ - ~ -~-~--. : : 

! 

Dietary ]Restriction, ~ T~ ~60 395 ] 

TA~L~ JSa 

Mean Body Weights an~ Surv~vaK of MKce in ~he D~e~ary ]Restriction S~udy 

of Scopo~an~ine Hyc~robro~n~de Tr~hydra~e: Ad Libitum Yeedin..g and We~gh~-Ma~ched Controls ]Protocols 

(continued) 

Ad L~/tum-Fed Weight-I~atched 

Weeks Control Control, 25 m~_.kg 

on Av. Wt.. No. of Av. Wt. No.0f Av. Wt. Wt. (% of Wt. (% of No. of 

Study (g) Survivors (g) Survivors (g) ad libitum-fed weight-matched Survivors. 

controls) controls) 

Female 

1 19.8 70 19.6 60 19.7 100 101 70 

2 21.4 69 19.6 59 20.9 98 107 70 

3 22.6 69 20.5 59 22.4 99 109 70 

4 23.9 69 21.6 59 23.2 97 107 70 

5 25.0 69 22.8 59 24.1 96 106 70 

6 25.6 69 22.8 59 24.4 95 107 70 

7 26.6 69 23.0 59 25.3 95 110 70 

8 28.2 69 24.6 59 26.2 93 107 70 

9 28.7 69 23.1 59 26.3 92 114 70 

10 29.6 69 23.8 59 26.5 90 111 70 

11 30.1 69 25.3 59 27.1 90 107 70 

12 30.7 69 24.6 59 27.6 90 112 70 

13 31.2 69 24.0 59 27.6 89 115 70 

17 34.8 69 25.3 59 29.3 84 116 70 

21 37.7 69 26.6 59 31.8 84 120 70 

25 38.7 69 30.1 59 32.1 83 107 70 

29 41.0 69 33.5 59 33.6 82 100 70 

33 43.5 69 31.0 59 35.1 81 113 70 

37 45.8 69 32.0 59 36.2 79 113 70 

41 47.0 69 34.7 59 37.9 81 109 70 

45 49.5 69 38.3 59 38.5 78 101 70 

49 51.0 69 39.2 59 39.1 77 100 70 

53 51.4 69 40.1 59 39.7 77 99 69 

57 52.0 68 41. I 59 39.5 76 96 69 

61 53.0 68 40.8 59 40.0 76 98 68 

65 54.0 68 40.5 59 41.1 76 102 67 

69 a 54.7 49 42.3 48 42.3 77 100 47 

73 56.0 48 42.4 48 42.0 75 99 47 

77 55.3 47 43.4 48 42.0 76 97 47 

81 53.9 47 42.2 48 41.4 77 98 45 

85 52.9 46 41.8 47 41.6 79 100 45 

89 53.3 40 40.8 44 42.3 79 104 44 

93 52.4 40 39.3 43 42.4 81 108 43 

97 50.2 39 39.7 43 41 .I 82 104 41 

101 47.9 38 37.6 42 40.3 84 107 38 

Mean for weeks 

1-13 26.4 22.7 24.7 94 109 

14-52 43.2 32.3 34.8 81 108 

53-101 52.8 40.9 41.2 78 101 

a Interim evaluation occurred during week 66. 
396 Dietary Restriction, NTP TR 460 

TABLE J5b 

Mean Body Weights and Survival of Mice in the Dietary Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-Year and 3-Year Restricted Feed Protocols 

2-Year Protocol 3-Year Protocol 

Weeks Vehicle Control 25 me/k~ Vehicle Control 25 mg/k~ 

on Av. Wt. No. of~ • Av. Wt. Wt. (% of No. of Av. Wt. No. of Av. Wt. Wt. (% of No. of 

Study (g) Survivors (g). controls) Survivors (g) Survivors (g) controls) Survivors 

Male 

1 24.7 60 24.7 100 60 24.4 50 24.1 99 50 

2 24.6 60 24.3 99 60 24.1 50 23.8 99 50 

3 23.9 60 23.7 99 60 24.5 50 23.9 98 50 

4 24.9 60 24.5 98 60 25.0 50 23.4 94 50 

5 26.9 60 25.8 96 60 26.6 50 25.4 96 50 

6~ 25.6 60 25.1 98 60 26.0 50 25.1 97 50 

7 23.8 60 23.1 97 60 23.3 50 23.2 100 50 

8 22.9 60 22.6 99 60 23.7 50 21.9 92 50 

9 22.7 60 23.2 102 60 24.8 50 22.7 92 50 

10 25.0 60 24.6 98 59 27.3 50 25.2 92 50 

It 24.2 60 23.3 96 59 " 25.2 50 23.6 94 50 

12 27.3 60 25.9 95 59 27.8 50 26.2 94 50 

13 28.4 60 27.2 96 59 27.9 50 26.2 94 50-

17 29.6 60 28.2 95 59 29.9 49 27.7 93 50 

21 30.0 60 28.2 94 59 30.3 49 27.8 92 50 

25 32.8 60 29.5 90 59 33.1 49 • 29.9 90 50 

29 30.4 60 27.6 91 59 30.3 49 27.2 90 50 

33 31.0 60 28.3 91 59 30.3 49 28.7 95 49 

37 30.8 60 29.1 95 59 31.3 49 28.8 92 49 

41 32.2 60 30.6 95 59 33.9 49 31.3 92 49 

45 32.4 60 30.3 94 59 33.6 49 30.2 90 49 

49 32.9 60 30.5 93 59 34.5 49 31.3 91 49 

53 34.2 60 '31.9 93 59 37.2 49 31.7 85 49 

57 34.7 ' 60 32.0 92 59 37.0 49 32.2 87 49 

61 32.2 60 29.8 93 59 34.7 49 29.9 86 49 

65 37.1 60 34.2 92 59 38.2 49 33.8 89 49 

"- 69 a 36.8 ,- 50 32.6 89 - 49 38.6 49 31.9 83 49 

73 38.7 50 34.1 88 48 39.5 49 33.3 84 49 

77 38.9 49 32.7 84 48 39.1 49 32.4 83 49 

81 36.8 49 31.1 85 48 36.7 49 30.8 84 49 

85 35.4 49 31.2 88 48 35.0 49 30.0 86 49 

89 35.2 49 31.3 89 48 34.7 48 30.4 88 49 

93 34.3 49 31.3 91 48 35.6 48 31.4 88 49 

97 33.9 49 30.4 90 48 32.8 48 29.8 91 49 

92 48 34.2 48 31.2 91 49 101 34.2 49 31.5 

34.2 48 31.2 91 49 105 . 

34.5 48 31.3 91 49 I09 

34.5 46 31.3 91 48 113 

35.4 45 31.6 89 48 I17 

35.9 45 31.9 89 47 121 

36.8 43 31.8 86 47 125 

36.7 42 31.6 86 ~ 129 

35.9 42 31.0 86 43 133 

34.4 40 31.1 90 39 137 

33.1 38 30.6 92 38 141 

34.2 36 32.1 94 38 145 

33.3 32 31.0 93 37 149 

33.6 28 31.3 93 37 153 " 

Mean for weeks 

1-13 25.0 24.5 98 25.4 24.2 95 

14-52 31.3 29.1 93 31.9 29.2 92 

53-101 35.6 31.9 90 36.4 31.4 86 

102-153 34.8 31.4 90 
Dietary Restriction, NTP TR 460 39// 

TABLE J5b 

Mean Body Weights and Survival of Mice in the Dietary ]Restriction Study 

of Scopolamine Hydrobromide Trihydrate: 2-'Year and 3,~rear ]Restricted Feed t~rotocols (continued) 

2-Year Protocol 3-Year Protocol 

Weeks Vehicle Control 25 m~.~k~ Vehicle Control 25 mg//k~ 

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. No. of Av. Wt. Wt. (% of No. of 

Study (g) Survivors (g) controls) Survivors (g) Survivors (g) controls) Survivors 

Female 

1 20.1 60 20.0 100 60 19.8 50 19.7 100 50 

2 20.6 60 20.2 98 60 20.2 50 19.6 97 50 

3 20.6 60 20.1 98 60 21.1 50 20.7 98 49 

4 22.3 60 21.1 95 60 21.4 50 20.5 96 49 

5 23.5 60 22.7 97 60 23.1 50 22.4 97 49 

6 23.0 60 22.2 97 60 22.9 50 22.4 98 49 

7 22.6 60 22.1 98 60 22.5 50 21.6 96 49 

8 22.9 60 22.1 97 60 22.2 50 21.5 97 49 

9 23.0 60 21.9 95 60 22.3 50 22.3 100 49 

10 24.2 60 23.5 97 60 24.3 50 23.6 97 49 

11 22.9 60 22.4 98 60 22.9 50 21.9 96 49 

12 24.9 60 24.3 98 60 25.0 50 24.0 96 49 

13 26.0 60 24.7 95 ~0 25.0 50 24.3 97 49 

17 26.8 60 25.6 96 60 26.5 50 25.1 95 49 

21 28.1 60 26.5 94 60 27.7 50 25.6 92 49 

25 29.6 60 28.3 96 60 30.2 50 27.5 91 49 

29 28.3 60 26.8 95 60 28.2 50 25.7 91 49 

33 29.2 60 28.3 97 60 29.9 50 27.8 93 49 

37 29.4 60 27.6 94 60 29.5 50 27.1 92 49 

41 30.2 60 28.8 95 60 32.2 50 28.4 88 49 

45 30.4 60 28.9 95 60 32.2 50 28.0 87 49 

49 31.0 60 29.5 95 60 32.6 50 29.5 91 49 

53 32.5 60 30.2 93 60 33.2 50 30.1 91 49 

57 31.8 60 30.5 96 59 32.5 50 .30.4 94 49 

61 30.3 60 28.7 95 58 31.4 50 28.6 91 49 

65 34.0 60 32.8 97 58 34.7 50 31.6 91 49 

69 a 33.0 50 30.8 93 48 33.8 50 31.0 92 49 

73 34.8 49 32.7 94 48 35.4 50 31.5 89 49 

77 34.5 49 32.1 93 48 35.0 50 31.0 89 49 

81 32.5 48 30.2 93 47 33.1 47 29.6 89 48 

85 31.4 48 29.8 95 47 31.9 47 28.5 89 48 

89 30.9 48 29.8 96 46 31.0 46 28.9 93 48 

93 30.5 47 29.4 96 45 31.8 46 29.4 93 48 

97 29.9 47 29.0 97 44 29.7 45 27.5 93 48 

101 30.8 47 29.9 97 44 31.4 45 29.1 93 45 

105 30.6 45 28.9 94 45 

109 31.0 44 28.8 93 43 

113 30.4 43 28.9 95 43 

117 31.0 42 28.8 93 39 

121 31.2 42 29.0 93 38 

125 30.4 37 29.0 95 34 

129 30.1 35 29.0 96 3~ 

133 29.6 29 28.8 97 33 

137 28.9 27 28.3 98 32 

141 28.0 25 27.7 99 28 

145 29.7 24 29.7 100 25 

149 28.7 23 28,8 100 24 

153 29.0 21 28.9 100 19 

Mean for weeks 

1-13 22.8 22.1 97 22.5 21.9 97 

14-52 29.2 27.8 95 29.9 27.2 91 

53-101 32.1 30.5 95 32.7 29.8 91 

102-153 29.9 28.8 96 

a Interim evaluation occurred during week 66 O f the 2-year protocol. 
398 Dietary Restriction, NTP TR 460 
3~9 

APIPEI ]]]}]IZ ]K 

FEED AlSr]I )C©MP©II..]IZ]I]) C© 5U IPTI©I 

JIM TIHIL., ]IETAI Y Z5T ]ICT]I©M 5TUJI])]IIZ5 

TABLE K~.a Feed and Compound Consumption by Rats 

an the Dietary Restriction Study of butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 400 

• TABLE ~lb Feed and Compound Consumption by Rats 

an the Dietary Restriction Study of butyl Benzyl Phthalate: 

2-Year and Long-Term Restricted Feed Protocols ............................ 402 

~ABLE ~.2a Feed and Compound Consumption by Rats 

an the Dietary Restriction Study of t-Butylhydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 404 

TABLE ~2b Feed and Compound Consumption by Rats 

an the Dietary Restriction Study of t-Butylhydroquinone: 

30-Month Restricted Feed Protocol .................................... 406 

TABLE ]K3a Feed Consumption by Male Rats 

an the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum "Feeding and Weight-Matched Controls Protocols ................... 408 

TABLE K3b Feed Consumption by Male Rats 

an the Dietary Restriction Study of Salicylazosulfapyridine: 

2-Year and 30-Month Restricted Feed Protocols . ........................... 409 

TABLE K4a Feed Consumption by Male Mice 

an the Dietary Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols ................... 410 

TABLE K4b Feed Consumption by Male Mice 

an the Dietary Restriction Study of Salicylazosulfapyridine: 

2-Year and 3-Year Restricted Feed Protocols .............................. 411 
400 Dietary Restriction, NTP TR 460 

o 

TABLE Kla " 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a . • 

Ad ILibitum- . , Weight-Matched 

Fed Control Control ~2,000 ppm 

Feed . Body' Feed Body Feed Body Feed/Control Feed e Dose/ 

Week 

(g/day) b Weight 

(g) (g/day) Weight (g) (g/day) Weight(g) . Ad LibitumFed Control 

WeightMatched Day d 

(mg/kg) 

Control 

Mal~ ~ ~_~ 

1 ~ 14.6 110 

2 14.6 149 15.5 146 14.1. 141 0.97 0.91 1,198 

3 15.9 183 16.7 181 

4 15.6 205 16.8 210 

5 17.8 233 16.7 234 19.7, 226 1.11 1.18 1,043 

6 17.5 253 16.9 252 18.4. 243 - 1.05 1.09 905 

7 16.5 266 

8 17.4 286 16.3 282 

9 16.5 298 17.0 297 15.6 284 0.95 0.92 660 

10 16.2 310 16.4 306 16.6 291 1.02 1.01 686 

11 15.3 319 16.4 319 

12 16.4 326 16.1 325 

13 16.4 • 339 16.4 335 16.8 315 1.02 1.02 642 

17 17.3 370 t4.3 365 12.3 338 0.71 0.86 438 

~ 21 15.9 391 13.6 352 17.5 351 1.10 1.29 599 

25 20.5 391 14.1 364 15.5 362 0.76 1.10 512 

29 14.7 416 14.2 381 14.4 377 0.98 1.01 458 

33 14.7 423 13.0 399- ~ 15.6 382 1.06 1.20 490 

37 , 15.0 429 10.4 395 15.2 389 1.01 1.46 469 

41 17.1 437 • 12.3 378 16.5 394 0.96 1.34 502 

45 17.5 443 12.4 391 17.0 405 0.97 1.37 503 

49 16.1 449 13.2 365 15,9 413 0.99 1.20 463 

-~'~53 16.3 451 14.3 " 400 16.7 414 1.02 1.17 484 

57~ 15.3 ~ 450 14.4 391 16.2 412 1.06 1.13 470 

61 - 15.7 451 i3.8 413 16.1 413 1.03 1.17 468 

65 15.4 45i 14.0. 412 15.1 420 0.98 1,08 431 

69 14.2 446 13.9- 422 15.1 419 1.06 1.09 434 

73 15.1 448 13.7 427 15.4 427 1.02 1.12 433 

77 14.1 446 13.7 • 427 14.4 428 1.02 1.05 404 

81 13.5 440 12.9 ' 432 13.3 422 0.99 1.03 378 

85 13.6 440 13.8 433 14.3 418 1.05 1.04 409 

89 13.9 439 12.1 438 13.6 413 0.98 1.12 396 

93 13.1 438 13.0 422 0.99 370 

97 14.5 433 12:5 434 12.7 411 0.88 1.02 372 

101 12.4 426 13.3 433 11.6 402 0.94 0.87 347 

• "-'°,Mean for weeks 

1-1~3 16.3 264 16.3 250 16.9 250 1.02 1.02 856 

14-52~. 16.5 417 13.1 377 15.5 379 0.95 1.20 493 

53-101 ~ 14.5 443 13.5 423 14.4 417 1.00 1.07 415 
i 

l 

Dietary Restriction, NTP TR 460 401 

TABLE Kla 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

. 

Ad Libitum- Weight-Matched 

Fed Control Control 24,000 ppm 

Feed Body Feed Body Feed Body Feed/C0ntrol Feed Dose/ 

(g/day) Weight (g/day) Weight (g/day) Weight Ad Libitum- Weight- Day 

Week (g) (g) (g) Fed Control Matched (mg/kg) 

Control 

Female 

1 11.0 97 

2 10.3 121 11.2 120 7.5 114 0.73 0.67 1,582 

3 11.0 135 11.2 134 

4 11.3 142 11.0 142 

5 11.1 153 11.1 151 

6 10.6 159 11.4 159 11.3 151 1.07 0.99 1,791 

7 10.9 167 11.8 166 

8 10.9 171 10.6 170 

9 11.3 175 10.3 174 

10 10.9 178 10.7 180 10.5 171 0.96 0.98 1,478 

11 10.8 180 10.5 183 

12 11.0 186 9.9 186 

13 10.3 185 10.3 184 9.8 176 0.95 0.95 1,338 

17 10.2 198 9.6 193 9.7 183 0.95 1.01 1,274 

21 10.3 204 10.4 203 9.9 187 0.96 0.95 1,269 

25 9.7 209 10.1 193 1.04 1,254 

29 9.3 218 8.5 221 9.0 197 0.97 1.06 1,090 

33 9.7 223 7.3 211 9.3 199 0.96 1.27 1,127 

37 10.7 231 5.8 204 9.6 203 0.90 1.66 1,i33 

41 10.7 238 6.6 198 

45 I 1.4 240 6.8 196 9.7 207 0.85 1.43 1,119 

49 12.5 258 7.4 194 10.4 212 0.83 1.41 1,177 

53 11.6 269 7.7 199 I0.1 214 0.87 1.31 1,130 

57 11.1 276 7.8 200 10.6 214 0.95 1.36 1,184 

61 11.4 281 7.6 202 9.7 216 0.85 .1.28 1,073 

65 11.7 286 7.4 203 10.1 220 0.86 1.36 1,104 

69 11.1 296 7.5 216 9.9 222 0.89 1.32 1,068 

73 11. I 298 7.5 208 9.8 226 0.88 1.31 1,046 

77 11.3 300 8.2 228 9.8 230 0.87 1.20 1,028 

81 11.7 300 7.2 228 10.0 222 0.85 1.39 1,084 

85 11.8 312 8.2 239 10.8 231 0.92 • 1.32 1,126 

89 12.1 321 8.1 245 10.6 237 0.88 1.31 1,074 

93 10.6 320 8.4 238 10.5 236 0.99 1.25 1,069 

97 11.4 319 9.1 247 10.4 233 0.91 1.14 1,070 

101 10.2 318 8.0 254 10.1 232 0.99 1.26 1,049 

Mean for weeks 

1-13 10.9 163 10.8 157 9.8 153 0.93 0.90 1,547 

14-52 10.6 226 8.0 203 9.7 198 0.92 1.23 1,180 

53-101 11.3 300 7.9 224 10.2 226 0.90 1.29 1,085 

a Feed consumption by controls was measured weekly for 13 weeks and monthly thereafter. 

b Grams of feed consumed per animal per day 

c Grams of feed consumed per exposed animal per day divided by grams of feed consumed per control animal per dayd Milligrams of butyl benzyl phthalate consumed per kilogram body weight per day 

~j!~~ 

i 
402 Dietary Restriction, NTP TR 460 

TABLE Klb 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of Butyl Benzyl Phthalate: 

2-Year and L0ng-Term Restricted Feed Protocols ~ 

2-Year Protocol 30-Month Prot0¢0] 

' 0 uum 12.000 m~m 0 prim 12.000 Dnm 

_. • Feed Body Feed Body Feed/ Dose/ Feed Body Feed Body Feed/ Dose/ 

...(g/day) a Weight (g/day) Weight Control Day c (g/day) Weight (g/day) Weight Control Day 

Week (g) (g) Feed b . (mg/kg) (g) (g) Feed (mg/kg) 

Male 

1 14.3 103 13.8 101 0.97 1,643 14.6 103 13.9 101 0.95 1,649 

2 12.2 140 11.6 138 0.95 1,013 12.1 139 ll.7 138 0.97 1,024 

3 11.9' 146 12.2 143 1.03 1,023 11.9 146 12.2 143 1.03 1,023 

4 12.6 156 12.7 155 1.01 " 983 12.5 155 12.6 154 1.01 979 

5 14.1 176 14.0 168 0.99 998 14.1 182 13.9 167 0.99 1,002 

6 13.8 200 14.1 180 1.02 943 14.0 204 14.1 179 1.01 944 

7 14.2 208 13.8 199 0.97 834 14.2 210 13.9 204 0.98 821 

8 14.9 227 15.3 216 1.03 850 14.8 233 15.3 217 1.03 843 

,9 13.3 246 13.5 234 1.02 690 13.0 246 14.4 244 I. I l 707 

10 13.3 260 13.5 248 1.02 652 13.2 262 13.8 254 1.05 655 

I 1 13.7 258 13.7 249 1.00 661 13.7 261 13.7 258 1.00 639 " 

•. 12 15.5 261 15.9 251 1.03 760 15.7 263 16.2 256 1.03 759 

13 13~6 285 14.0 274 1.03 615 13.6 287 14;2 280 1.04 611 

17 14.6 •301 14.8 294 1.01 604 14.7 307 14.9 303 1.01 589 

21 . 14.5 339 • 14.5 324 1.00 537 14.7 349 14.4 323 0.98 533 

, .!!5 13.3 358 13.8 339 1.04 488 13.3 ' 363 13.7 342 1.03 480 

29 13.1 349 13.1. 340 1.00 461 13.0 356 13.2 344 1.02 459 

!;3 12.8 362 12.9 347 1.01 447 12.8 376 13.0 351 1.02 444 

37 13.1 366 13.0 344 0.99 454 13.1 378 13.2 346 '1.01 457 

,41 13.0 383 12.9 349 0.99 443 12.9 390 12.9 351 1.00 442 

45 12.0 372 12.2 345 1.02 426 II.9 376 ' 12.3 • 357 1.03 413 

49 12.5 366 12.3 343 0.98 432 12.4 371 12.4 344 1.00 434 

53 " 13.8 368 13.7 351 0.99 468 13.8 376 • 13.6 357 0.99 457 

57 13.4 378 13.5 370 1.01 439 13.4 398 13.4 373 1.00 430 

61 • 13.5 399 13.5 375 1.00 432 13.3 404 13.5 378 1.02 427 

($5 13.3 388 13.4 374 1.01 430 13.2 405 13.4 379 1.02 425 

69 12.1. 406 12.2 377 1.01 388 12.1 408 12.1 381 1.00 380 

73 10.9 403 10.7 378 0.98 339 10.8 410 10.8 374 1.00 346 

77 9.4 377 9.6 354 1.02 325 9.4 376 11.9 347 1.27 413 

81 .." 11.2 395 11.1 372 0.99 356 11.2 389 11.0 364 0.98 364 

85 9.8 398 9.8 362 1.00 325 9.9 399 9.9 364 1.00 327 

89. 10.3 371 10.8 342 1.05 380 9.9 371 10.9 345 1.10 379 

93 10.1 365 10.0 362 0.99 332 I0. I 378 10.2 367 1.01 334 

97 10.2 371 10.3 341 1.01 361 I0.2 386 10.3 348 . 1.01 354 

101 10.2 354 10.0 340 0.98 353 I0.3 370 10.3 342 1.00 360 

105 I0.I 360 10.3 334 1.02 369 

109 " I0.2 359 IOA 327 0.99 371 

113 9.8 359 10.2 332 1.04 368 

117 lO.O 332 10.2 314 1.02 389 

1'21 9.9 324 10.1 304 1.02 400. 

1:25 I0.0 329 10.2 307 1.02 397" 

Mean for weeks 

1-13 . 13.6 205 13.7 197 1.00 898 13.6 207 13.8 200 1.01 897 

14-52 13.2 355 13.3 336 1.00 477 13.2 363 13.3 340. 1.01 472 

53-I01 11.4 383 11.4 .361 1.00 379 11.4 390 11.6 363 1.03 384 

102q25, 10.0 344 10.2 320 1.02 382 
. . . . . ~,.,.~_~_ ........ ~ T - ~ o ~ - ~ = : ~ = ~ = ~ - ...... --~---~--~:.:-.~:77 ..... ~_,~ , - _ : ......... : 

J 

Dietary Restriction, NT~ TR 460 403 

i~ 

TABLE Klb 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of Butyl Benzyl.Phthalate: 

2-'Year and Long-Term Restricted Feed Protocols (continued) 

2-year Protocol 32-Month Protocol 

0 ~pm 24,000 ppm 0 ppm 24,000 ppm ,,, 

Feed Body Feed. Body Feed/ Dose/ Feed Body Feed Body, Feed/ Dose/ 

(g/day) Weight (g/day) Weight Control Day (g/day) Weight (g/day) Weight Control Day 

Week (g) (g) Feed (mg/kg) (g) (g) Feed (mg/kg) 

Female 

1 11.1 94 7.7 88 0.69 2,105 11.4 95 8.4 87 0.74 2,302 

2 11.6 116 12.0 107 1.03 2,684 11.7 118 12.0 105 1.03 2,749 

3 12.0 128 11.9 118 0.99 2,411 12.1 128 11.8 ' 116 0.98 2,437 

4 9.1 131 8.9 124 0.98 1,715 9.2 131 8.8 122 0.96 1,730 

5 8.1 133 8.0 127 0.99 1,515 8.1 134 8.1 127 1.00 1,519 

6 8.0 146 8.0 134 1.00 1,435 8.0 146 8.0 133 1.00 1,447 

7 8.1 141 8.1 130 1.00 1,494 8.2 142 8.1 130 0.99 1,493 

8 8.3 141 8.3 141 1.00 1,421 8.3 141 8.4 139 1.01 1,450 

9 9.1 159 9.0 148 0.99 1,465 9.1 158 9.0 147 0.99 1,473 

10 8.3 155 8.2 145 0.99 1,358 8.4 155 8.3 145 0.99 1,387 

11 8.0 157 8.1 , 146 1.01 1,323 8.0 157 8.1 146 1.01 1,328 

12 8.0 160 8.1 152 1.01 1,272 8.0 161 8.1 151 1.01 1,280 

13 8.1 162 8.0 155 0.99 1,247 8.1 163 8.2 151 1.01 1,311 

17 8.1 170 8.2 164 1.01 1,197 8.1 170 8.2 164 1.01 1,195 

21 8.0 176 8.1" 164 1.01 1,182 8.0 180 8.0 164 1.00' 1,180 

25 8.0 189 8.0 178 1.00 1,081 8.0 188 8.0 176 1.00 1,086 

29 7.7 193 7.9 180 1.03 1,047 7.7 194 8.4 180 1.09' 1,115 

33 7.0 194 6.9 175 0.99 954 7.0 195 6.9 173 0.99 957 

37 7.2 182 7.3 176 1.01 989 7.3 183 7.3 180 1.00 969 

41 7.9 191 7.8 175 0.99 1,073 7.9 199 7.8 173 0.99 1,085 

45 8.6 190 9.1 180 1.06 1,214 8,6 192 9.1 178 1.06 1,225 

49 8.6 198 8.6 184 1.00 1,114 8.5 . 200 8.5 184 1.00 1,104 

53 I0.0 206 9.3 191 0.93 1,168 10.0 209 9.2 191 0.92 1,154 

57 10.4 217 9.6 194 0.92 1,183 10.4 221 9.2 191 0.88 1,159 

61 10.2 222 9.5 198 0.93 1,157 10.4 227 9.6 196 0.92 1,175 

65 10.0 228 .9.3 197 0.93 1,126 10.2 232 9.1 197 0.89 1,109 

69 10.4 233 9.9 202 0.95 . 1,175 9.8 238 9.8 199 1.00 1,184 

73 10.0 244 9.0 208 0.90 1,038 10.4 247 9.0 203 0.87 1,065 

77 10.9 254 9.2 208 0.84 1,058 10.5 257 9.0 207 0.86 1,041 

81 11.1 263 9.5 210 0.86 1,086 11.0 269 9.3 . 205 0.85 1,084 

85 10.6 273 10.0 211 0.94 1,136 10.7 280 9.8 206 0.92 " 1,145 ' 

89 11.0 276 9.6 212 0.87 1,094 10.9 284 9.4 207 0.86 1,083 

93 11.0 282 10.0 212 0.91 1,130 10.7 289 10.1 210 0.94 1,161 

97 9.9 284 9.7 211 0.98 1,109 10.1 294 9.4 210 0.93 . 1,078 

101 10.2 276 10.0 213 0.98 1,127 10.2 293 10.0 209 0.98 1,147 

105 10.1 288 9.7 210 0.96 1,116 

109 10.1 290 10.0 211 0.99 1,138 

113 9.6 283 9.9 211 1.03 1,134 

117 9.9 283 9.8 210 0.99 1,123 

121 10.1 282 10.0 208 0.99 1,155 

125 9.9 291 9.6 206 0.97 1,124 

129 10.0 288 9.6 203 0.96 1,139 

133 10.2 294 9.1 198 0.89 .1,101 

137 10.3 286 9.6 189 0.93 1,227 

Mean for weeks 

1-13 9.1 140 8.8 132 0.98 1,650 9.1 141 8.9 131 0.98 1,685 

14-52 7.9 187 8.0 175 1.01 1,094 7.9 189 8.0 175 1.01 1,102 

53-I01 10.4 251 9.6 205 0.92 1,122 I0.4 257 9.5 202 0.91 1,122 

102-137 I0.0 287 9.7 205 0.97 1,140 

J : 

i , 

a Grams of feed consumed per animal per day 

b Grams of feed consumed per exposed animal per day divided by grams of feed consumed per control animal per day 

c Milligrams of butyl benzyl phthalate consumed per kilogram body weight per day 
404 Dietary Restriction, NTP TR 460 

TABLE K2a 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of t-B, utylhydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a .. , 

Ad ILibitum- Weight-Matched 

Fed Control Control 5,000 nnm 

Feed Body Feed Body Feed Body Feed/Control Feed c Dose/ 

(g/day) b Weight (g/day) Weight (g/day) Weight Ad Libitum- Weight- Day d 

Week (g) (~ (g) Fed Control Matched (mg/kg) 

Control 

Male 

1 13.9 99 14.3 97 

2 15.2 141 14.3 139 15.7 127 1.03 1.10 619 

3 16.5 175 13.0 161 

4 18.2 209 13.4 188 

5 17.4 233 14.0 189 

6 20.8 267 16.3 215 18.3 244 0.88 1.12 375 

7 17.2 283 14.6 247 

8 17.0 299 14.0 250 

9 16.9 313 15.1 260 

10 13.1 316 15.7 273 17.5 299 1.34 1.11 293 

11 19.2 324 14.9 296 

12 17;4 344 15.0 314 

13 16.8 356 16.1 323 17.6 320 1.05 1.09 275 

17 16.5 379 14.9 364 17.2 346 1.04 1.15 248 

21 16.4 400 13.0 336 17.2 358 . 1.05 1.32 241 

25 15.6 413 13.4 348 15.9 376 1.02 1.19 211 

29-~- 16.5 419 14.1 384 16.4 386 0.99 1.16 213 

,33 16.4 428 13.8 380 16.7 392 1.02 1.21 213 

37 17.4 437 13.3 389 16.4 402 0.94 1.23 204 

41 14.7 450 13.4 393 15.2 416 1.03 1.13 182 

45 16.5 452 14.0 406 16.9 415 1.02 1.21 203 

49 16.9 444 13.7 398 16.0 415 0.95 1.17 193 

53 19.9 468 13.8 417 20.8 431 1.05 1.51 242 

57 14.8 468 14.7 415 16.8 435 • 1.14 1.14 193 

61, 16.1 467 14.7 419 17.7 440 1.10 1.20 202 

65 16.3 470 14.0 429 17.6 440 1.08 1.26 200 

69 15.8 468 14.1 428 16.7 437 1.06 1.18 191 

73 15.3 472 13.2 423 16.0 433 1.05 1.21 185 

• 77 ~ 17.0 463 13.6 435 16.3 438 0.96 1.20 186 

81 16.6 464 14.3 432 17.3 436 1.04 1.21 199 

~ 85 15.4 462 13.8 428 15.1 429 0.98 1.09 176 

89 15.1 455 13.6 431 14.0 425 0.93 1.03 164 

93 14.3 458 13.3 429 15.1 419 1.06 1.14 180 

97 15.4 455 13.7 434 16.0 i 421 1.04 1.17 190 

101 15.2 447 12.8 430 15.7 421 1.03 1.23 186 

105 14.5 440 13.4 426 16.4 409 1.13 1.22 201 

109 15.7 420 13.6 416 14.2 412 0.90 1.04 173 

113 14.8 429 12.9 419 15.7 395 1.06 1.22 198 

117 14.0 428 9.9 399 15.0 397 1.07 1.52 189 

121 14.4 417 10.7 406 15.5 386 1.08 1.45 200 

Mean for weeks 

1-13 16.9 258 14.7 227 • 17.3 247 1.07 1.11 390 

1,1-52 16.3 425 13.7 378 16.4 390 1.01 1.20 212 

53-121 15.6 453 13.3 . 423 16.2 422 " 1.04 1.22 192 
:~ ~z:,~=e~:- ~ ~ 7-~.-~. ~ - - ~ .... ~_ -____~_~ _._:~:~--~ - - ..... . ::_ 

• 

Dietary Restriction, NTP TR 460 405 1 

TABLE ]~2a 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

Ad Libitum Feeding and Weight-Matched Controls Protocols (continued) 

Ad Libitum- Weight-Matched 

Fed Control Control 5~000 ppm I:: !~ 

Feed Body Feed Body Feed Body Feed/Control Feed Dose/ 

Week 

(g/day) Weight

(g) 

(g/day) Weight

(g) 

(g/day) Weight

(g) 

Ad ILibitumFed Control 

WeightMatched Day 

(mg/kg) i 

Control i; 

i 

i : 

Female 

1 10.8 92 11.1 92 

2 11.3 121 10.9 " 121 11.9 110 1.05 1.09 541 

3 11.3 135 7.3 126 

4 11.6 147 7.5 130 

5 11.I 159 7.4 123 

6 13.7 171 8.2 129 11.3 154 0.82 1.38 367 

7 11.6 177 12.0 155 

8 10.8 181 7.3 161 

9 10.7 186 7.2 149 

10 9.4 176 7.9 150 11.1 176 I. 18 1.41 316 

11 11.9 193 11.7 168 

12 10.5 196 9.6 175 

13 10.2 198 9.8 180 11.2 184 1.10 1.14 304 

17 10.7 203 10.6 192 9.8 188 0.92 0.92 262 

21 10.7 224 8.8 192 10.8 200 1.01 1.23 270 

25 9.4 222 8.7 207 9.5 203 1.01 1.09 235 

29 9.2 229 8.8 213 10.1 210 1.10 1.15 240 

33 10.1 231 8.7 210 10.0 210 0.99 1.15 239 

37 10.9 232 8.5 219 10.4 213 0.95 1.22 244 

41 I0.1 240 8.6 227 9.1 219 O. 90 1.06 207 

45 10.8 248 8.6 225 11.0 227 1.02 1.28 242 

49 11.6 257 8.7 231 10.7 232 0.92 1.23 232 

53 15.3 264 8.7 227 14.1 236 0.92 1.62 299 

57 9.6 279 9.4 236 10.8 246 1.13 1.15 219 

61 11.3 285 9.4 240 11.2 257 0.99 1.19 217 

65 12.2 292 9.2 243 11.5 263 0.94 1.25 219 

69 11.6 301 9.6 245 10.7 265 0.92 1.11 202 

73 12.2 309 9.2 250 11.0 270 0.90 1.20 204 

77 12.4 315 8.9 259 11.5 274 0.93 1.29 209 

8P 12.3 323 10.4 257 11.8 278 0.96 I. 13 213 

85 11.5 327 9.8 266 11.3 282 0.98 i. 15 200 

89 11.7 333 10.3 272 10.9 287 0.93 1.06 190 

93 12.3 342 10.0 274 11.1 290 0.90 1.11 191 

97 12.6 348 10.3 270 12.5 299 0.99 1.21 209 

101 12.0 347 10.8 282 11.9 301 0.99 1.10 197 

105 13.0 345 10.9 285 12.3 303 0.95 1.13 203 

109 12.5 341 10.5 287 12.0 300 0.96 1.14 200 

113 11.1 337 12.1 295 12.4 301 1.12 1.02 205 

117 11.7 348 10.5 295 11.4 309 0.97 1.09 184 

121 12.3 341 10.6 294 12.0 315 0.98 1.13 190 

125 I 1.9 336 10.5 289 12.0 307 1.01 1.14 196 

Mean for weeks 

1-13 11.2 164 9.1 143 11.4 156 1.04 1.25 382 

14-52 10.4 232 8.9 213 10.2 211 0.98 1.15 241 

53-125 12.1 322 10.0 267 11.7 283 0.97 1.17 208 

a Feed consumption by controls was measured weekly for 13 weeks and monthly thereafter. 

b Grams of feed consumed per animal per day 

e Grams of feed consumed per exposed animal per day divided by grams of feed consumed per control animal per day 

d Milligrams of t-butylhydroquinone consumed per kilogram body weight per day 
406 Dietary Restriction, NTP TR 460 

TABLE K2b 

Feed and Compound Consumption by Rats in the Dietary Restriction Study of t-Butylhydroquinone: 

30,-Month Restricted Feed Protocol 

0 ppm 5,000 ppm 

Feed Body Feed Body Feed/ Dose/Day c 

(g/day) a Weight (g/day) Weight Control (mg/kg) 

Week (g) (g) Feed b 

Male 

1 14.3 . 107 13.8 94 0.97 732 

2 16.2 151 13.2 139 0.81 476 

3 14.1 185 12.0 167 0.85 360 

' 4 12.7 185 12.6 163 0.99 387 

5 12.2 195 12.0 179 0.98 334 

6 13.1 208 12.8 195 0.98 329 

7 14.1 231 14.4 216 1.02 333 

8 14.2 246 14.2 229 1,00 310 

9 14.0 255 14.2 240 1.01 295 

10 14.2 266 14.5 250 1.02 290 

11" 14.7 274 14.8 260 1.01 286 

12 14.1 289 14.0 276 0.99 254 

13 14.8 291 15.1 281 1.02 270 

17 14:7 314 14.3 309 0.97 232 

21 15.8 351 15.6 341 0.99 229 

25 14.6 343 14.4 342 0.99 211 

29 14.4 348 14.6 344 1.01 212 

33 14.4 358 14.5 362 1.01 200 

37 14.5 371 14.6 370 1.01 196 

41 14.0 390 14.0 387 1.00 182 

45 14.7 404 14.7 397 1.60 185 

49 13.1 403 13.3 393 1.02 170 

53 12.6 386 12.7 391 1.01 162 

57 12.1 392 12.2 394 1.01 155 

61 12.4 395 12.3 391 0.99 158 

65 12.0 398 12.4 396 1.03 156 

69 12.9 399 13.0 395 1.01 165 

73 12.4 395 12.0 389 0.97 154 

77 12.3 382 12.4 378 1.01 165 

81 13.1 393 13.3 393 1.02 169 

85 12.7 395 12.8 390 1.01 164 

89 12.8 401 12.6 397 0.98 158 

93 12.3 400 12.1 391 0.98 155 

97 11.9 403 11.7 389 0.98 151 

101 12.0 401 12.2 387 1.02 158 

1,05 13.1 369 12.6 364 0.96 173 

109 11.9 383 11.9 372 1.00 161 

H3 11.4 370 10.9 367 0.96 149 

H7 11.6 364 11.1 348 0.96 159 

~.21 12.1 360 11.4 348 0.94 164 

125 11.6 350 11.5 343 0.99 168 

i29 11.3 324 11.1 315 0.98 177 

Mean for weeks 

1-13 14.0 222 13.7 - 207 0.97 358 

14-52 ' 14.5 365 14.5 361 1.60 202 

53-129 12.2 383 12.1 377 0.99 161 
Dietary Restriction, NTP TR 460 407 

I 

TABLE K2b 

Feed and Compound Consumption by Rats in ~he Dietary Restriction Study of t-Buty~hydroquinone: ! 30-1V~onth ~estr~cted Feed ~Protocol (continued) I: 

i 

0 hum , 5.000 ~¢m 

Feed Body Feed Body Feed/ Dose/Day 

(g/day) Weight (g/dayi Weight Control (mg/kg)

Week (g) (g) Feed 

Female 

1 11.4 92 11.0 82 0.96 669 

2 11.7 128 11.2 120 0.96 465 

3 8.8 129 8.2 120 0.93 341 

4 7.5 128 7.8 120 1.04 323 [ 5 7.6 132 7.3 124 0.96 294 

6 7.1 134 7.3 129 1.03 283 

7 7.7 141 7.6 135 0.99 281 

8 7.9 146 7.8 141 0.99 279 

9 8.4 150 7.2 145 0.86 251 

10 8.3. 153 7.6 144 0.92 264 

I1 8.5 157 8.0 148 0.94 273 

12 8.7 162 8.0 155 0.92 257 

13 9.1 181 8.9 175 .0.98 255 

17 8.3 172 8.1 171 0.98 236 

21 8.7 189 8.8 188 1.O1 233 

25 8.8 181 8.8 183 1.00 241 

29 8.5 184 8.4 183 0.99 230 

33 8.9 194 9.0 195 l.Ol 230 

37 8.9 212 9.0 213 1.01 210 

41 8.1 213 7.7 214 0.95 180 

45 7.7 208 7.6 210 0.99 181 

49 7.4 209 7.5 211 l.Ol 178 

53 7.9 204 7.9 204 1.00 192 

57 8.6 221 8.7 224 1.01 195 

61 9.0 224 8.8 226 0.98 196 

65 9.3 231 9.2 235 0.99 196 

69 10.1 236 10.3 237 1.02 217 

73 10.4 240 10.5 243 1.01 216 

77 10.4 244 10.5 244 1.01 216 

81 12.2 254 12.2 258 1.00 237 

85 11.5 264 11.2 262 0.97 214 

89 12.2 279 11.6 274 0.95 212 

93 12.2 294 11.9 283 0.98 211 

97 12.1 302 11.5 290 0.95 198 

101 11.9 308 11.5 296 0.97 195 

105 12.4 317 11.8 289 0.95 204 

109 11.2 ~15 11.2 295 1.00 189 

113 10.6 317 10.4 296 0.98 176 

117 9.7 308 9.0 285 0.93 158 

121 10.0 300 9.4 278 0.94 169 

125 9.5 285 9.1 273 0.96 166 

129 8.9 270 9.1 264 1.02 172 

Mean for weeks 

1-13 8.7 14! 8.3 134 0.96 " /326. 

14-52 8.4 196 8.3 196 0.99 213 

53-129 10.5 271 10.3 263 0.98 196 

a Grams of feed consumed per animal per day 

b Grams of feed consumed per exposed animal per day divided by grams of feed consumed per control animal per day 

c Milligrams of t-butylhydroquinone consumed per kilogram body weight per day 
408 Dietary Restriction, NTP TR460 

TABLE K3a ~ ~ ~, 

Feed Consumption by Male Rats in the Dietary Restriction Study of Salicylazosulf~pyridine: (

Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad ILibitum- : Weight-Matched 

Fed Control Control 337.5 m~/k~ 

Feed Body Feed Body Feed Body Feed/Control FeedC. 

•. (g/day) b ~ Weight (g/day) Weight (g/day). Weight Ad Libitum- WeightWeek (g) (g) (g) • Fed Control Matched 

- " Control _ 

I 12.9 

2 15.1 

3 13.6 

4 15.9 

5 15.6 

6 14.8 

7 15.6 

8 

9 15.2 

10 14.6 

11 14.1 

12 14.2 

13 15.4 

14 13.3 

18 14.2 

21 15.1 

25 15.5 

29 13.5 

33 13.3 

37 13.1 

41 13.7 

45 12.7 

49 12.3 

53 15.8 

57 8.2 

61 I1.0 

65 12.3 

" 69 15:4 

73 11.9 

77 

81 10.9 

85 9.7 

89 9.9 

93 8.8 

97 9.0 

101 . 8.0 

Mean forweeks 

1-13 14.7 

14-52 13.7 

53-101 10.9 

102 13.9 i03 12.8 102 0.99 ". 0.92 

139 14.4 136 14.9 135 0.99 1.03 

171 15.8 174 

196 15.7 200 

216 15.1 220 14.4 215 0.92 0.93 

238 15.7 243 8.4 240 0.57 0.54 

258 15.7 259 

14.9 273 

28~ 14.3 286 14.0 280 0.92 0.9~ 

299 15.0 298 18.5 296 1.27 1.23 

307 14.6 305 

317 14.0 316 

324 14.5 324 13.2 316 0.86 0.91 

333 14.5 337 15.0 331 1.13 1.03 

360 13.0 364 18.5 358 1,30. 1.42 

374 13.5 371 13.2 363 0.87 0.98 

~93 16.0 394 13.9 386 0.90 0.87 

416 14.3 410 13.2 400. 0.98 . 0.922 

424 12.6 425 11.7 413 0.88 0~93 

439 13.7 431 11.6 423 0.89 0.85 

447 14.1 437 12~2 428 0.89 0.87 

457 13.7 451 11.2 439 0.88 0.82 

461 11.9 459 11.4 444 0.93 0.96 

469 !1.9 .463 13.2 451 0.84 I.'11 

471 11.7 467 8.3 460 1,01 0.71 

477 12.5 468 10.2 453 0.93 0.82 

477 11.1 474 11.1 459 0.90 1.00 

474 12.9 471 10.5 461 0.68 , 0.81 

477 12.5 470 12.9 463 1.08 1.03 

12.5 469 13.7 451 1.i0 

468 11.7 468 11.5 449 - 1.06 0.98 

467 11.8 458 9.0 450 0.93 0.76 

461 10.7 460 11.5 444 1.16 1.07 

453 10.5 449 12.4 439 1.41 1.i8 

442 9.8 437 12.0 437 . 1.33 .1.22 

422 9.4 426 15.3 417 1.9i 1.63 

238 14.9 ~241 13.8 226 0.93 0.94 

410 13.7 408 13.2 398 0.96 0.96 

463 H.5 460 11.7 449 1.10 1.03 

:. 

a Feed consumption by controls was measured weekly for 13 weeks and monthly thereafter. 

b Grams o~ feed consumed per animal per day 

c Grams Of feed consumed per dosed animal per day divided by grams of feed consumed per control animal per day. 

.. 
. . . . . . . . ~:-~;;~'~-'::5::~'-- ~ < ~ .... ~:7'~'-~: : :'-7:~: ....... i 

! 

[ 

i

!: 

\~ ! Dietary Restriction, NTP TR 460 ~ 

i ~ 

i 

TABLE K3b 

~eed Consumption by Male Rats in the Dietary Restriction Study of Salicylazosulfapyridine: 

2-'Year and 30-Month Restricted ~eed Protocols .. i~ 

2-Year Protocol 30-Month Protocol 

, ,.Ve, hicig Control 337.5 m~/k~ Vehicle Control 337.5 m~/_~l~ 

Feed Body Feed Body . Feed/ Feed Body Feed Body Feed/ 

(g/day) a Weight (g/day) Weight Control (g/day) Weight (g/day) Weight Control 

Week (g) Feed b (g) (g) Feed 

= 

I 13.4 107 13.4 108 1.00 13.9 109 13.3 109 0.96 

2 il.6 140 11,5 140 0.99 11.6 143 11.4 141 0.98 

3 11.9 167 11,9 167 1.00 11.9 171 11.9 168 1.00 

4 12.6 183 12",5 181 0,99 12.6 183 12.6 181 1.00 

5 12.8 205 12,8 205 1.00 12.7 204 12.8 204 1.01 

6 12.1 216 12,0 215 0,99 12.1 217 12.1" 217 1.00 

7 12.2 230 12.3 230 1.01 12.3 232 12.3 231 1.00 

8 12.1 241 12.1 240 1,00 12.2 243 12.1 238 0.99 

9 12.2 250 12.2 248 1.00 12.3 253 12.3 245 1.00 

10 12.3 261 12.1 255 0.98 12.1 261 12.2 254 1.01 

11 12.0 270 11.9 267 0.99 12.3 273 11.8 264 0.96 

12 11.9 283 11,6 281 0.97 11.9 289 11.7 277 0.98 

14 12.1 298 11.9 296 0.98 12.2 302 12.0 290 0.98 

17 11.1 306 I1.0 298 0.99 11.2 315 11.0 295 0.98 

20 11.1 318 10.9 308 0.98 10.8 320 10.7 306 0.99 

24 11.3 336 11.1 324 0.98 11.4 339 11,5 319 1,01 

28 11.4 345 11.4 325 1.00 11.4 344 11.4 324 1.00 

32 10.9 359 11.3 341 1.04 "10.9 357 11.1 339 1.02 

37 li.2 363 11.3 343 1.01 11.3 363 11.2 347 0.99 

40 11.3 370 11.3 354 1.00 11.1 375 11.3 351 1.02 

44 11.2' 383 11.1 356 0.99 11,1 381 11.2 357 1.01 

48 11.0 384 I1.I 359 1.01 11.2 387 11.1 362 0.99 i 

53 11.8 383 11.9 360 1.01 11,8 384 11.9 361 1.01 [ 

56 11.6 408 10.8 382 0.93 11.6 408 10.9 385 0.94 

60 12.1 420 11.6 387 0.96 12.0 422 11.8 388 0.98 

64 11.2 425 11.4 382 1.02 11.5 424 11.3 381 0.98 

68 11.1 427 10.8 ~ 386 0.97 11.1 433 10.8 386 0.97 

72 10,3 423 10.3 378 1.00 10.3 425 10.3 378 1.00 

76 9.8 409 9.6 361 0.98 9.6 408 9.7 361 1.01 

80 9.8 403 9.7 349 0.99 9.7 407 9,8 348 1.01 

84 9.8 405 9.8 345 1.00 9.7 401 9.8 348 1.01 

89 9.9 402 9.8 342 0.99 9.8 400 9.9 346 1.01 

92 9.7 396 9,9 340 1.02 '9.7 394 9,9 341 1.02 

96 10.0 400 10.0 339 1.00 10.0 398 9.9 340 0.99 

100 9,7 400 9.7 329 1.00 9,4 386 9.3 329 0.99 

104 8,5 384 8.5 317 1.00 8.6 388 8.5 ' 327 0.99 

108 8.1 375 8.4 311 1.04 

112 8.5 380 8.3 304 0.98 

116 8.1 365 8.1 295 1.00 

120 8,1 351 8.0 282 0.99 

124 8.1 335 8.0 273 0.99 

128 8.2 304 8.1 255 0.99 

. 

Mean for weeks 

1-13 12.3 213 12.2 211 0.99 12.3 215 12.2 . 211 0.99 

14-52 11.2 346 11.2 330 1.00 11.3 348 11.2" "329 1.00 

53-104 10.4 406 10.3 357 0.99 10.3 405 10.3 358 0.99 

105-128 8.2 352 8.2 286 1.00 

a Grams of feed consumed per animal per day 

b Grams of feed consumed per dosed animal per day divided by grams of feed consumed per control animal per day 
410 Dietary Restriction, NTP TR 460 

TABLE K4a 

Feed Consumption by Male Mice in the Dietary, Restriction Study of Salicylazosulfapyridine: 

Ad Libitum Feeding and Weight-Matched Controls Protocols a 

Ad Libitum- Weight-Matched 

Fed Control Cgntrol 2,700 mg/kg 

Feed , Body Feed Body Feed Body Feed/Control Feed ¢ 

(g/day) b Weight (g/day) Weight (g/day) Weight Ad Libitum- WeightWeek (g) (g) (g) Fed Control Matched 

Control 

1 5.3 23.4 

2 5.7 25.3 5.1 24.7 6.6 25.0 1.16 1.29 

3 5.3 25.9 . 5.5 26.0 

4 5.6 27.7 5.4 27.3 

5 5.7 28.4 5.5 28.0 

6 6.0 29.1 5.5 28.5 6.5 28.3 1.08 1.18 

7 5.2 30.0 5.7 29.0 

8. 7.3 31.4 5.8 30.2 

9 4.6 31.6 5.2 31.0 

10 5.9 32.0 5.4 31.0 7.3 30.8 1.24 1.35 

11 5.7 32.9 5.2 . 31.9 

12 5.0 33.5 5.2 32.3 

13 5.8 34.6 4.7 33.7 

14 5.2 35.2 4.7 33.5 6.3 32.5 1.21 1.34 

18 5.6 38.2 5.2 36.5 6.1 34.2 1.09 1.17 

22 5.8 40:7 5.3 40.1 7.0 37.0 1.21 1~32 

26 5.6 44.7 5.9 42.4 6.6 37.9 1.18. 1.12 

30 5.7 45.2 3.5 37.5 6.3 39.2 1.11 1.80 

34 5.3 47.7 4.0 35.3 6.3 39.4 1.19 1.58 

38 5.1 48.7 4.6 39.6 6.6 40.0 1.29 ! .43 

42 4.8 49.2 4.7 41.7 5.6 40.1 1.17 1.19 

46 4.9 49.7 4.3 ' 45.1 6.4 41.0 1.31 1.49 

50 4.9 50.2 '3.9 42.2 6.0 41.8 1.22 1.54 

54 4.8 51.7 4.0 43.6 5.5 43.1 1.15 1.38 

58 5.0 52.6 3.8 45.3 5.9 44.7 1.18 1.55 

62 4.4 52.8 3.8 47.7 5.7 45.9 1.30 1.50 

66 5.7 53.3 6.7 46.2 1.18 

68 3.9 47.1 

70 5.5 54.0 3.9 46.8 7.2 45.2 1.31 1.85 -

74 5.8 53.6 4.0 45.1 8.3 48.2 1.43 2.08 

78 5.2 54.4 3.8 45.3 6.8 45.8 1.31 1.79 

82 5.2 54:.3 3.9 43.8 7.1 47.2 1.37 1.82 

86 5.0 52.7 4.2 45.4 6.4 46.2 1.28 1.52 

90 ' 4.5 52.9 4.0 49.0 .6.1 46.8 1.36 1.53 

94 4.4 53.4 3.9 49.5 5.7 47.2 1.30 1.46 

98 4.3 52.3 4.0 49.9 5.5 44.8 1.28 1.38 

102 4.8 52.0 3.6 50.5 6.7 45.7 1.40 1.86 

Mean for weeks 

1-13 5.7 30.2 5.3 29.0 6.8 28.0 1.16 1.28 

14-52 5.3 45.0 4.6 39.4 6.3 38.3 1.20 1.40 

53-102 5.0 53.1 3.9 46.9 6.4 45.9 1.29 1.64 

a Feed consumption by controls was measured weekly for 13 weeks and monthly thereafter. 

b Grams of feed consumed per animal per day 

c Grams of feed consumed per dosed animal per day divided by grams of feed consumed per control animal per day 
i .......................... ~...................... ~ - ~ .... ~ - ~ ~ , , ~ ~ ..... ~ . . . . . ~ ~ ~ : ~ : ~ 

Diet.ary Restriction, NTP TR 460 411 

TABLE K4b 

Feed Consumption by Male Mice in the Dietary Restriction Study of Salicy]azosulfapyridine: 

2-~'ear and 3-Year Restricted Feed Protocols 

2-Year Protocol 3-Year Protocol 

~ehicle Control 2,700 m.g/k.~ Vehicle Control 2,700 m~k~ _ 

Feed Body Feed Body • Feed/ Feed Body Feed Body Feed/ 

(g/day) a " Weight (g/day) Weight Control (g/day) Weight (g/day) Weight Control 

Week (g) (g) Feed b (g) (g) Feed 

1 3.7 23.4 4.8 23.2 1.30 3.7 23.7 4.8 23.5 

2 3.7 25.1 3.8 24.7 1.03 3.7 25.3 3.9 25.3 

3 3.0 24.9. 3.0 24.3 1.00 3.0 24.9 3.0 24.7 

4 3.1 23.4 3.1 22.8 1.00 3.1 23.3 3.1 23.1 

5 3.3 24.9 3.3 23.5 1.00 3.3 24.7 3.3 23.9 

6 3.5 24.9 3.6 23.2 1.03 3.5 24.7 3.6 23.4 

7 3.5 25.9 3.5 23.7 1.00 3.5 25.6 3.5 23.9 

8 3.5 27.0 3.5 25.2 1.00 3.5 27.0 3.5 24.9 

9 3.7 27.1 3.7 25.5 1.00 3.7 26.8 3~7 25.8 

10 3.6 27.9 3.6 26.1 1.00 3.7 27.9 3.6 26.5 

I 1 3.6 27.8 3.6 26.5 1.00 3.6 27.7 3.6 26.7 

12 3.6 29.0 3.6 26.2 1.00 3.6 29.0 3.6 26.4 

13 3.6 29.4 3.6 27.2 1.00 3.6 29.3 3.6 27.4 

17 3.6 32.4 3.6 28.3 1.00 3.6 32.0 3.6 28.5 

18 3.6 32.9 3.7 28.6 1.03 3.6 32.4 3.7 28.7 

22 3.6 35.0 3.6 30.1 1.00 3.6 34.4 3.6 30.3 

26 3.9 35.8 3.9 31.4 1.00 3.9 35.3 3.9 31.6 

30 4.3 40.7 4.4 32.6 1.02 4.3 39.9 4.5 33.1 

34 4.3 44.7 4.4 34.5 1.02 4.4 43.5 4.4 34.8 

38 3.3 46.0 3.6 33.8 1.09 3.3 45.1 3.5 33.6 

42 3.2 42.5 4.0 , 33.8 1.25 3.2 41.7 4.0 33.8 

46 3.5 41.1 4.0 34.1 1.14 3.5 40.4 4.0 34.4 

50 3.6 40.4 4.0 32.7 1.11 3.6 39.7 4.0 32.7 

54 4.5 42.6 4.6 33.8 1.02 4.6 42.0 4.6 33.6 

58 4.3 49.6 4.3 36.6 1.00 4.3 48.9 4.4 36.8 

62 3.7 49.5 3.6 35.8 0.97 3.7 48.5 3.6 36.0 

66 3.6 48.9 3.6 34.9 1.00 3.5 47.5 3.6 34.7 

70 3.5 47.8 3.6 34.6 1.03 3.6 46.4 3.6 34.5 

74 3.4 46.9 3.3 34.5 0.97 3.4 45.5 3.3 34.2 

78 3.3 45.0 3.3 33.6 1.00 3.3 43.9 3.3 33.6 

82 3.5 43.9 3.5 32.2 1.00 3.6 42.9 3.5 32.1 

86 3.6 41.8 3.6 31.0 1.00 3.6 40.4 3.6 31.2 

90 4.0 44.1 3.9 34.7 0.98 4.0 42.5 3.9 34.7 

94 3.9 45.8 4.0 34.8 1.03 3.9 44.3 3.9 34.4 

98 4.5 47.6 4.6 34.8 1.02 4.5 45.2 4.6 34.6 

102 3.8 49.4 3.9 35.8 1.03 3.8 46.5 3.9 35.2 

106 3.5 46.7 3.5 35.0 

110 3.6 46.5 3.5 35.6 

114 3.6 46.0 3.7 35.2 

118 3.6 45.3 3.7 33.6 

122 3.6 43.8 3.6 33.4 

126 3.7 43.0 3.7 34.0 

130 3.7 41.3 3.7 33.2 

134 3.6 41.4 3.6 33.6 

138 3.3 39.3 3.3 32.5 

142 3.5 39.3 3.5 32.8 

146 3.5 38.5 3.6 32.8 

150 3.6 36.2 3.6 30.5 

154 3.4 35.7 3.4 31.8 

Mean for weeks 

1-13 3.5 26.2 3.6 24.8 1.03 3.5 26.1 3.6 25.0 

14-52 3.7 39.2 3.9 32.0 1.07 3.7 38.4 3.9 32.2 

53-102 3.8 46.4 3.8 34.4 1.00 3.8 45.0 3.8 34.3 

103-154 3.6 41.8 3.6 33.4 

1.30 

1.05 

1.00 

1.00 

1.00 

1.03 

1.00 

1.00 

1.00 

0.97 

1.00 

1.00 

1.00 

1.00 

1.03 

1.00 

1.00 

1.05 

1.00 

1.06 

1.25 

1.14 

1.11 

1.00 ° 

1.02 

0.97 

1.03 

1.00 

0.97 

1.00 

0.97 

1.00 

0.98 

1.00 

1.02 

1.03 

1.00 

0.97 

1.03 

1.03 

1.00 

1.00 

1.00 

1.00 

1.00 

1.00 

1.03 

1.00 

1.00 

1.03 

1.06 

1.00 

1.00 

[ 

a Grams of feed consumed peranimal per day 

b Grams of feed consumed per dosed animal per day divided by grams of feed consumed per control animal per day ' 

l 
DEPARTMENT OF 

HEALTH & HUMAN SERVICES 

SPECIAL FOURTH-CLASS RATE 

Public Health Service POSTAGE AND FEES PAID 

National Toxicology Program 

Central Data Management 

DHHS/NIH 

Permit No. G-811 

P.O. Box 12233, MD El-02 

Research T~iangle Park, NC 27709 

Official Business 

Penalty for Private Use - $300 

NIH Publication No. 97-3376 

September 1997