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A  Carcinogenicity  Bioassay  of 
Isobutyl  2-Cyanoacrylate  (IBC)  in 
Fischer-344  Rats  —  Final  Report 


Larry  D.  Brown,  DVM,  LTC,  VC 
Paul  W.  Mellick,  DVM,  COL,  VC 
Catherine  D.  Smith,  DVM,  MAJ,  VC 
and 

Don  W.  Korte,  Jr.,  PhD,  LTC,  MSC 


DISTRIBUTION  STATEMENT  A 

Approver  fsi  public  release} 
Duaicuuct  ’laiumted 


MAMMALIAN  TOXICOLOGY  BRANCH 
DIVISION  OF  TOXICOLOGY 


January,  1990 


Toxicology  Series:  187 


LETTERMAN  ARMY  INSTITUTE  OF  RESEARCH  PRESIOFO  OF  SAN  FRANCISCO  CALIFORNIA  94129 


\  Carcinogenicity  Bioassay  of  Isobutyl  2-Cyanoacrylate  (IBC)  in  Fisclier-344  Rats  -  Final 
Report  (Toxicology  Series  187)- 
Brown  el  at.  . ... .  ...  ,  i 


This  document  has  been  approved  for  public  release  and  sale;  its  distribution  is 
unlimited. 

Destroy  this  report  when  it  is  no  longer  needed.  Do  not  return  to  the  originator. 

Citation  of  trade  names  in  this  report  does  not  constitute  an  official  endorsement 
or  approval  of  the  use  of  such  items. 

This  research  was  conducted  in  compliance  with  the  "Guide  for  the  Care  and  Use 
of  Laboratory  Animals,"  NIH  Publication  No.  85-23,  as  prepared  by  the  Institute 
of  Laboratory  Animal  Resources,  National  Research  Council. 


This  material  has  been  reviewed  by  Letterman  Army  Institute  of 
Research  and  there  is  no  objection  to  its  presentation  and/or 
publication.  The  opinions  or  assertions  contained  herein  are  the 
private  views  of  the  author(s)  and  are  not  to  be  construed  as  official 
or  as  reflecting  the  views  of  the  Department  of  the  Army  or  the 
Department  of  Defense.  (AR  360-5) 


Donald  G.  Corby 
COL,  MC 
Commanding 


c AjT&K^O 


(date) 


BEST 

AVAILABLE  COPY 


URITY  CLASSIFICATION  OF  This  PAGE 


REPORT  DOCUMENTATION  PAGE 


Form  Approved 
OMB  No.  0704-0188 


la  REPORT  SECURITY  CLASSIFICATION 
UNCLASSIFIED 


2a.  SECURITY  CLASSIFICATION  AUTHORITY 


2b.  DECLASSIFICATION  /DOWNGRADING  SCHEDULE 


4  PERFORMING  ORGANIZATION  REPORT  NUM3ER(S) 


Institute  Report  No.:  443 


6«.  NAME  OF  PERFORMING  ORGANIZATION 

Mammalian  Toxicology 
Division  of  Toxicology 


6c.  ADDRESS  (City,  State,  end  ZIP  Code) 

Letterman  Army  Institute  of  Research 
Presidio  of  San  Francisco,  CA  94129-680 


6b.  OFFICE  SYMBOL 
(If  Applicable) 

SGRD-ULE-T 


lb  RESTRICTIVE  MARKINGS 


3  DISTRIBUTION /AVAILABILITY  OF  REPORT 

APPROVED  FOR  PUBLIC  PF LEASE; 
DISTRIBUTION  IS  UNLIMITED. 


5  MONITORING  ORGANIZATION  REPORT  NUMBER(S) 


7a.  NAME  OF  MONITORING  ORGANIZATION 

US  Army  Institute  of  Dental  Research 


7b.  ADDRESS  (City.  State,  and  ZIP  Code) 


Washington,  DC  20307-5400 


8a.  NAME  OF  FUNDING /SPONSORING 

organization  us  Army  Medical 

Research  &  Development  Corr 


8c  AOORESS  (City,  State,  and  ZIP  Code) 

Fort  Detrick 

Frederick,  Maryland  21701-5012 


1 1 .  TITLE  (include  Security  Classification) 

(U)  A  Carcinogenicity  Study  of 
Rats  —  Final  Report 


12.  PERSONAL  AUTHOR(S) 

_ LD  Brown,  PW  Mellick, 


13a.  TYPE  OF  REPORT  |1 3b.  TIME  COVERED 


PROCUREMENT  INSTRUMENT  IDENTIFICATION  NUMBER 


10.  SOURCE  OF  FUNDING  NUMBERS 


PROGRAM 
ELEMENT  NO. 

62775A 


Isobutyi  2-Cyanoacrylate  (IBC)  in  Fischer-344 


Institute 


16.  SUPPLEMENTARY  NOTATION 


from  30JAN8&O  14FEB8 


CD  Smith,  and  DW  Korte.  Jr, _ 


14.  DATE  OF  REPORT  (Year,  Month,  Day) 


January  1990 


15.  PAGE  COUNT 

1150 


COSATI  COOES 


GROUP  SUB-GROUP 


Toxicology  Series  No.  187 


18.  SUBJECT  TERMS  ( Continue  on  reverse  if  necessary  and  identify  by  block  number) 

Chronic  Toxicity/  Isobutyl  2-Cyanoacrylate,  IBC, 
Bucrylate®,-,’Mammalian  Toxicology,  Tissue 
Adhesive;  Carcinogenicity  Bioassav/  Rat^  ;  - 


19.  ABSTRACT  < Continue  on  reverse  if  necessary  and  identify  by  block  number) 

This  report  covers  the  results  from  the  second  year  of  a  two-year 
^ 1 inogen i c i t y  bioassay  of  the  tissue  adhesive,  isobutyl  2— cyanoacrylate 
(IBC).  Four  hundred  seven  6-week-old  Fischer-344  rats  were  randomized  into 
three  groups  (control,  10  p.1  IBC,  and  100  Jil  IBC),  each  group  containing  both 
male  and  female  animals.  ^The  IBC  was  administered  by  surgical  implantation  of 
the  liquid  monomer  directly  onto  the  ventral  capsule  of  the  liver.  The 
monomer  was  allowed  to  polymerize  before  two-layer  closure  of  the  abdominal 
incision.  Control  animals  received  100  ^1  of  isotonic  saline,  also  by 
surgical  implantation.  All  animals  were  examined  daily,  and  weighed  and 
palpated  monthly.  „A  dose-related  increase  in  the  incidence  of  clinically 
observed  intra-abdominal  masses  was  evident  among  IBC-treated  animals  of  both 
sexes.  In  addition  to  the  masses  and  the  more  permanent  sequelae  of  the 
surgical  procedure  (xyphoid  protuberance,  corneal  opacity) ,  the  animals 


20  DISTRIBUTION /AVAILABILITY  OF  ABSTRACT 
□tUNCLASSIFIEO/UNLIMITED  □  SAME  AS  RPT 


22a.  NAME  OF  RESPONSIBLE  INDIVIDUAL 
DONALD  G.  CORBY,  COL.  M 


DO  Form  1473,  JUN  86 


21  ABSTRACT  SECURITY  CLASSIFICATION 
□  DTic  USERS  UNCLASSIFIED 


22b  TELEPHONE  (Include  Area  Code) 

(415)  561-3600 


Previous  editions  are  obsolete. 


SECURITY  CLASSIFICATION  OF  THIS  PAGE 


19  (cont.)  ^presented  with  a  variety  of  transitory  clinical  signs  sporadically 
throughout  the  first  and  second  year.  These  signs  were  observed  in  all  dose 
groups,  and  could  not  be  attributed  to  compound  administration.  The  weight 
gain  of  the  two  IBC  treatment  groups  was  comparable  to  that  recorded  for  the 
control  group. 


Three  hundred  five  animals  remained  on  study  at  the  start  of  the  second 
year.  During  the  second  year,  118  animals  died  or  were  sacrificed 
prematurely.  Twenty-one  were  unscheduled  deaths  and  97  were  unscheduled 
sacrifices.  At  the  end  of  the  second  year,  the  187  remaining  animals  were 
sacrificed.  All  animals  (118  unscheduled  and  187  scheduled)  were  evaluated  by 
necropsy  during  the  second  year.  One  hundred  eighty-four  of  the  animals  that 
received  the  IBC  had  fibrotic  adhesions  between  the  liver  and  the  omentum, 
peritoneal  membrane,  diaphragm,  stomach,  skin,  and/or  intestine.  These 
lesions  were  characterized  histologically  as  foreign  body  granulomatous 
reactions.  Thirty-one  rats  treated  with  IBC  developed  sarcomas  during  the 
second  year.  The  first  sarcoma  was  detected  on  day  399  after  intra-abdominal 
administration  of  the  test  material.  Twenty-one  of  31  animals  with  sarcoma 
died  or  were  sacrificed  in  a  moribund  condition,  while  10  animals  with  sarcoma 
survived  until  terminal  sacrifice.  Twenty-seven  of  31  animals  with  sarcoma 
had  grossly  detectable  masses  at  necropsy.  Four  sarcomas  were  detected  only 
by  microscopic  examination.  The  liver  was  the  most  frequently  affected  organ^^ 
(27  of  31  animals  had  lesions  in  the  liver) .  Four  IBC-treated  rats  (2  low- 
dose  males  and  2  high-dose  females)  had  hepatocellular  carcinomas  of  the 
liver,  while  controls  had  none.  F-344  leukemia  or  large  granular  lymphocyte 
leukemia  (LGLL)  occurred  in  62  animals  distributed  among  all  groups,  including 
controls.  Solid-state  carcinogenesis  mechanisms  are  the  most  probable  cause 
for  the  sarcomas,  but  humans,  unlike  rodents,  fail  to  elicit  a  solid-state 
carcinogenesis  effect.  The  incidence  of  hepatocellular  carcinomas  was  twice 
that  reported  for  rats  of  the  same  strain  and  age,  which  provides  reason  for 
concern.  However,  the  incidence  of  this  tumor  was  not  statistically 
significant  and  there  was  no  dose-response  pattern  by  sex.  All  other  gross  or 
histopathological  findings  were  considered  incidental  to  IBC  treatment,  or 
were  sequelae  of  the  surgical  procedure. 


Results  of  this  study  indicate  that  the  ??  sence  of  IBC  in  the  abdominal 
cavity  had  no  effect  on  survival,  weight  gain,  jr  the  clinical  condition  of  F- 
344  rats  during  the  second  year  following  its  implantation.  IBC  did  produce 
sarcomas  in  the  abdomen  of  16%  of  the  animals.  The  sarcomas  were  attributed 
to  a  solid-state  effect  which  is  not  present  in  man.  A  nonstatistically 
significant  increase  in  hepatocellular  carcinomas  was  observed  in  4  IBC- 
treated  rats  but  there  was  no  clear  evidence  that  this  could  be  attributed  to 
the  IBC  implants.  ,,  o  j  ^  p 


"7' 


.-p 


ABSTRACT 


This  report  covers  the  results  from  the  second  year  of  a 
two-year  carcinogenicity  bioassay  of  the  tissue  adhesive, 
isobutyl  2-cyanoacrylate  (IBC).  Four  hundred  seven  6-week- 
old  Fischer-344  rats  were  randomized  into  three  groups 
(control,  10  |il  IBC,  and  100  ^.1  IBC)  ,  each  group  containing 
both  male  and  female  animals.  The  IBC  was  administered  by 
surgical  implantation  of  the  liquid  monomer  directly  onto  the 
ventral  capsule  of  the  liver.  The  monomer  was  allowed  to 
polymerize  before  two-layer  closure  of  the  abdominal 
incision.  Control  animals  received  100  |il  of  isotonic 
saline,  also  by  surgical  implantation.  All  animals  were 
examined  daily,  and  weighed  and  palpated  monthly.  A  dose- 
related  increase  in  the  incidence  of  clinically  observed 
intra-abdominal  masses  was  evident  among  IBC-treated  animals 
of  both  sexes.  In  addition  to  the  masses  and  the  more 
permanent  sequelae  of  the  surgical  procedure  (xyphoid 
protuberance,  corneal  opacity) ,  the  animals  presented  with  a 
variety  of  transitory  clinical  signs  sporadically  throughout 
the  first  and  second  year.  These  signs  were  observed  in  all 
dose  groups,  and  could  not  be  attributed  to  compound 
administration.  The  weight  gain  of  the  two  IBC  treatment 
groups  was  comparable  to  that  recorded  for  the  control  group. 

Three  hundred  five  animals  remained  on  study  at  the 
start  of  the  second  year.  During  the  second  year,  118 
animals  died  or  were  sacrificed  prematurely.  Twenty-one  were 
unscheduled  deaths  and  97  were  unscheduled  sacrifices.  At 
the  end  of  the  second  year,  the  187  remaining  animals  were 
sacrificed.  All  animals  (118  unscheduled  and  187  scheduled) 
were  evaluated  by  necropsy  during  the  second  year.  One 
hundred  eighty-four  of  the  animals  that  received  the  IBC  had 
fibrotic  adhesions  between  the  liver  and  the  omentum, 
peritoneal  membrane,  diaphragm,  stomach,  skin,  and/or 
intestine.  These  lesions  were  characterized  histologically 
as  foreign  body  granulomatous  reactions.  Thirty-one  rats 
treated  with  IBC  developed  sarcomas  during  the  second  year. 
The  first  sarcoma  was  detected  on  day  399  after  intra¬ 
abdominal  administration  of  the  test  material.  Twenty-one  of 
31  animals  with  sarcoma  died  or  were  sacrificed  in  a  moribund 
condition,  while  10  animals  with  sarcoma  survived  until 
terminal  sacrifice.  Twenty-seven  of  31  animals  with  sarcoma 
had  grossly  detectable  masses  at  necropsy.  Four  sarcomas 
were  detected  only  by  microscopic  examination.  The  liver  was 
the  most  frequently  affected  organ  (27  of  31  animals  had 
lesions  in  the  liver) .  Four  IBC-treated  rats  (2  low-dose 
males  and  2  high-dose  females)  had  hepatocellular  carcinomas 
of  the  liver,  while  controls  had  none.  F-344  leukemia  or 
large  granular  lymphocyte  leukemia  (LGLL)  occurred  in  62 
animals  distributed  among  all  groups,  including  controls. 


i 


Solid-state  carcinogenesis  mechanisms  are  the  most  probable 
cause  for  the  sarcomas,  but  humans,  unlike  rodents,  fail  to 
elicit  a  solid-state  carcinogenesis  effect.  The  incidence  of 
hepatocellular  carcinomas  was  twice  that  reported  for  rats  of 
the  same  strain  and  age,  which  provides  reason  for  concern. 
However,  the  incidence  of  this  tumor  was  not  statistically 
significant  and  there  was  no  dose-response  pattern  by  sex. 

All  other  gross  or  histopathological  findings  were  considered 
incidental  to  IBC  treatment,  or  were  sequelae  of  the  surgical 
procedure . 

Results  of  this  study  indicate  that  the  presence  of  IBC 
in  the  abdominal  cavity  had  no  effect  on  survival,  weight 
gain,  or  the  clinical  condition  of  F-344  rats  during  the 
second  year  following  its  implantation.  IBC  did  produce 
sarcomas  in  the  abdomen  of  16%  of  the  animals.  The  sarcomas 
were  attributed  to  a  solid-state  effect  which  is  not  present 
in  man.  A  nonstatist ically  significant  increase  in 
hepatocellular  carcinomas  was  observed  in  4  IBC-treated  rats 
but  there  was  no  clear  evidence  that  this  could  be  attributed 
to  the  IBC  implants. 

Key  Words:  Chronic  Toxicity,  Isobutyl  2-Cyanoacrylate,  IBC, 
Bucrylate®,  Mammalian  Toxicology,  Tissue  Adhesive, 
Carcinogenicity  Bioassay,  Rat 


ii 


PREFACE 


TYPE  REPORT:  Chronic  Carcinogenicity  Bioassay  GLP  Final 
Report 

TESTING  FACILITY: 

US  Army  Medical  Research  and  Development  Command 
Letterman  Army  Institute  of  Research 
Presidio  of  San  Francisco,  CA  94129-6800 

SPONSOR:  US  Army  Medical  Research  and  Development  Command 
US  Army  Institute  of  Dental  Research 
Washington,  D.C.  20307-5400 

Project  Officer:  Eric  S.  Koppelman,  COL,  DC 

PROJECT/WORK  UNIT/APC:  Tissue  Adhesive  Project, 

35162775A825,  WU  015,  APC  TL11 

GLP  STUDY  NUMBER:  83009 

STUDY  DIRECTOR:  LTC  Don  W.  Korte,  Jr.,  PhD,  MSC 

Diplomate,  American  Board  of  Toxicology 

PRINCIPAL  INVESTIGATOR:  LTC  Larry  D.  Brown,  DVM,  MPVM,  VC 

Diplomate,  American  College  of 
Veterinary  Preventive  Medicine; 

Dipl.,  American  Board  of  Toxicology 

PATHOLOGISTS:  COL  Paul  W.  Mellick,  DVM,  PhD,  VC 

Diplomate,  American  College  of  Veterinary 
Pathology  (ACVP) 

MAJ  Catherine  D.  Smith,  DVM,  VC 
Diplomate,  ACVP 

TOXSYS™/DATA  MANAGER:  Yvonne  C.  LeTellier,  BS 

REPORT  AND  DATA  MANAGEMENT: 

A  copy  of  the  final  report,  study  protocol,  retired 
SOPs,  raw  data,  analytical,  stability,  and  purity 
data  of  the  test  compound,  tissues,  microslides, 
and  an  aliquot  of  the  test  compound  will  be 
retained  in  the  LAIR  Archives. 

TEST  SUBSTANCE:  Isobutyl  2-Cyanoacrylate  (IBC),  Bucrylate® 

INCLUSIVE  STUDY  DATES  (YEAR  2)  :  30  January  1985  - 

14  February  1986 

OBJECTIVE:  The  objective  of  this  study  was  to  evaluate  the 

carcinogenic/tumorigenic  potential  of  isobutyl  2- 
cyanoacrylate  in  male  and  female  Fischer-344  rats 
subjected  to  lifetime  (2-year)  exposure  to  the 
implanted  test  material. 


iii 


ACKNOWLEDGMENTS 

Richard  A.  Spieler,  RLAT  (AALAS) ,  SGT  Thomas  W.  Johnson, 
and  Nancy  J.  Smith  assisted  in  the  study.  SSG  James  D. 

Justus  and  staff  provided  animal  and  facility  management. 
Dianna  B.  Johnson  provided  office  management  and  secretarial 
assistance  for  this  study  and  preparation  of  the  report.  LTC 
William  G.  Rodkey,  DVM,  Diplomate,  American  College  of 
Veterinary  Surgeons,  COL  William  B.  Carpenter,  DDS,  and  LTC 
Eric  S.  Koppelman,  DDS,  performed  the  requisite  surgery  in 
support  of  this  project.  Histology  technicians  were  Lucille 
Cote  and  Marge  Henderson.  CPT  Dan  Brogdon,  DVM,  Ocular 
Hazards  Division,  was  the  veterinary  ophthalmology  consultant 
to  this  study.  CPT  Gary  M.  Zaucha,  DVM,  collated  the  final 
report.  CPT  Robert  E.  Harris,  DVM,  provided  assistance  with 
collating  the  final  report. 


PAUL  W.  MELLI^K,  DVM  /  DATE 
COL,  VC 

Senior  Pathologist 


C.  DAHLEM  SMITH,  DVM  /  DATE 
MAJ,  VC 

Assistant  Study  Pathologist 


DEPARTMENT  OF  THE  ARMY 


LETTERMAN  ARMY  INSTITUTE  OF  RESEARCH 
PRESIDIO  OF  SAN  FRANCISCO.  CALIFORNIA  94129-6800 

ATTENTION  OF; 

SGRD-ULZ-QA  19  January  1990 

MEMORANDUM  FOR  RECORD 

SUBJECT:  GLP  Compliance  for  GLP  Study  83009 


1.  This  is  to  certify  that  in  relation  to  LAIR  GLP  Study  83009 
the  following  inspections  were  made: 


23  August  1983 
17  January  1984 
23  January  1984 
17  February  1984 
26  March  1984 
17  May  1984 
05  July  1984 
02  August  1984 
29  January  1986 


Protocol  Review 

Weighing 

Weighing/Dosing 

Observation/Weighing 

Observation 

Observation 

Observation/Weighing 

Weighing 

Observation/Necropsy 


2.  The  institute  report  entitled  "A  Carcinogenicity  Bioassay  of 
Isobutyl  2 -Cyanoacrylate  (IBC)  in  Fischer-344  Rats — Final 
Report,"  Toxicology  Series  187,  was  audited  on  2  January  1990. 


to 

CAROLYN  M.  LEWIS 
Diplomate,  American  Board  of 
Toxicology 

Quality  Assurance  Auditor 


vi 


TABLE  OF  CONTENTS 


Volume  1 

Page 

Abstract . i 

Preface . iii 

Acknowledgments . iv 

Signatures  of  Principal  Scier  .ists . v 

Report  of  Quality  Assurance  Unit . vi 

Table  of  Contents . vii 

INTRODUCTION . 1 

Objective  of  Study . 1 

MATERIALS . 1 

Test  Substance . 1 

Animal  Data . 2 

Husbandry . 2 

METHODS . 3 

Group  Assignment/Acclimation . 3 

Dosage  Levels . 3 

Preparation  of  Compound . 3 

Chemical  Analysis  of  IBC . 3 

Test  Procedures . 4 

Clinical  Observations . 4 

Pathological  Examinations . 5 

Statistical  Analyses . 6 

Duration  of  Study . 6 

Changes/Deviations  from  Protocol . 6 

Storage  of  Raw  Data  and  Final  Report . 6 

RESULTS . 7 

Mortality . . . 7 

Clinical  Observations . 16 

Gross  Pathology . 23 

Microscopic  Pathology . 24 

DISCUSSION . 31 


vii 


TABLE  OF  CONTENTS  (cont.) 


Page 


CONCLUSION . 34 

REFERENCES . 35 

APPENDICES . 38 

Appendix  A.  Chemical  Data . 39 

Appendix  B.  Animal  Data . 4  6 

Appendix  C.  Surgical  Report . 48 

Appendix  D.  Historical  Listing  of  Study  Events . 54 

Appendix  E.  Glossary  of  Terms  for 

Clinical  Observations . 57 

Appendix  F.  Individual  Animal  Histories . 64 

Appendix  G.  Gross  Necropsy  Observations, 

Incidence  Summary . 369 

Appendix  H.  Microscopic  Observations, 

Incidence  Summary . 37  9 

Appendix  I .  Glossary  of  Terms  for 

Pathology  Findings . 402 

Appendix  J.  Pathology  Individual  Animal  Data . 466 

OFFICIAL  DISTRIBUTION  LIST . 1150 


viii 


A  Carcinogenicity  Bioassay  of  Isobutyl  2-Cyanoacrylate 
(IBC)  in  Fischer-344  Bats  --  Final  Report--Brown  et  al. 


INTRODUCTION 

Isobutyl  2-cyanoacrylate  (IBC)  is  being  evaluated  by  the 
U.S.  Army  Medical  Department  as  a  tissue  adhesive  for  use  in 
sutureless  wound  closure,  oral  and  maxillofacial  surgery,  and 
in  other  surgical  procedures.  The  use  of  this  modality  could 
provide  a  time-saving  and  sometimes  life-saving  dimension  to 
the  management  of  combat  wounds.  The  U.S.  Army  Institute  of 
Dental  Research  (USAIDR)  has  been  assigned  the  mission  of 
evaluating  the  therapeutic  potential  of  IBC.  As  part  of 
their  mandate,  USAIDR  has  tasked  the  Toxicology  Branch, 
Letterman  Army  Institute  of  Research  (LAIR) ,  to  evaluate  IBC 
in  a  chronic  carcinogenicity  bioassay. 

Results  from  the  first  year  of  the  study  indicate  that 
IBC  has  no  effect  on  survival,  weight  gain,  or  the  clinical 
condition  of  rats  during  the  first  year  following  its 
implantation.  The  only  gross  or  histopathological  finding 
observed  in  the  first  year  that  was  attributed  to  the  IBC 
treatment  was  the  presence  of  adhesions  and  a  granulomatous 
reaction  of  the  liver  and  those  adjacent  organs  that  came  in 
contact  with  the  IBC.  No  tumors  were  observed  during  the 
first  year  that  could  be  attributed  to  IBC  treatment  (1) . 

This  report  will  review  salient  features  of  the  first  year 
and  address  findings  that  occurred  during  the  second  year  of 
this  study. 

Objective  of  Study 

The  objective  of  this  study  was  to  evaluate  the 
carcinogenic/tumorigenic  potential  of  isobutyl  2- 
cyanoacrylate  in  male  and  female  Fischer-344  rats  subjected 
to  lifetime  (2-year)  exposure  of  the  implanted  test  material. 


MATERIALS 

Test  Substance 

Chemical  Name:  Isobutyl  2-cyanoacrylate  (IBC), 
Bucrylate® 

Chemical  Abstract  Service  Registry  No.:  1069-55-2 
Molecular  Formula:  C8HnN02 


Brown  et  al.  —  2 


Molecular  Structure: 

/COOH 
CH—C^ 


Additional  information  on  the  test  substance  is 
presented  in  Appendix  A. 

Animal  Data 

Two  hundred  nine  male  and  210  female  4-week-old  Fischer- 
344  (CDF)  rats  were  received  for  this  study  on  11  Jan  84  from 
Charles  River  Breeding  Laboratories,  Inc.,  Wilmington,  MA. 
They  were  identified  individually  with  ear  tags  numbered 
84D00001  to  84D00209  (inclusive)  for  the  males  and  84D00226 
to  84D00435  (inclusive)  for  the  females.  Four  males  and  4 
females  were  selected  randomly  for  quality  control  necropsy 
evaluation  at  receipt.  The  animal  weights  on  the  day 
following  receipt  (12  Jan  84)  ranged  from  30-57  g.  During 
quarantine,  three  underweight  females  and  one  maloccluded 
male  were  submitted  to  necropsy  on  20  Jan  84.  Additional 
animal  data  are  presented  in  Appendix  B. 

Husbandry 

Animals  used  in  this  study  were  housed  at  LAIR  in  the 
Toxicology  Suite,  a  restricted  access  facility.  Rats  were 
caged  individually  in  stainless  steel  wire-mesh  cages  in 
racks  equipped  with  automatically  flushing  dumptanks.  No 
bedding  was  used  in  any  of  the  cages.  The  diet,  fed  ad 
libitum,  consisted  of  Purina  Certified  Rodent  Chow®  Diet  5002 
(Ralston  Purina  Company,  St.  Louis,  MO).  Tap  water  was 
provided  by  automatic  water  valves  on  a  central  line.  Water 
was  analyzed  quarterly  for  impurities,  bacteria,  physical  and 
chemical  properties,  organic  residues,  pesticides,  and  heavy 
metals.  The  animal  room  temperature  and  relative  humidity 
were  continuously  recorded.  The  room  was  maintained  at 
temperatures  ranging  from  20.0°C  to  25.5°C,  with  a  relative 
humidity  range  of  35%  to  60%,  except  for  short  periods 
(spikes)  during  which  cleaning  of  the  room  altered  the 
relative  humidity,  and  on  six  occasions  when  increases  up  to 
70-86%  occurred  for  4-14  hours  due  to  steam  outages  or 
servicing  of  the  ventilation  system  fans.  The  photoperiod  of 
12  hours  of  fluorescent  light  per  day  was  electronically 
controlled-  Air  changes,  cage  size,  and  husbandry  conformed 
to  National  Research  Council  Institute  of  Laboratory  Animal 
Resources  standards  (2) .  The  LAIR  animal  facility  is 


CH, 

I  3 

HC — CHj 
CH3 


Brown  et  al.--3 


accredited  by  the  American  Association  for  Accredited 
Laboratory  Animal  Care. 


METHODS 

This  study  was  performed  in  accordance  with  the 
protocol,  applicable  amendments,  and  cited  operating 
procedures  including:  LAIR  Standard  Operating  Procedure  0P- 
STX-81,  "Chronic  Bucrylate  Bioassay  Procedures  within 
Toxicology  GLP  Suite  and  Administrative  Areas"  (3) ;  OP-STX- 
73,  "Chronic  Carcinogen  Bioassay"  (4);  and  FDA  Nonclinical 
Laboratory  Studies,  Good  Laboratory  Practice  Regulations  (5) . 

Group  Assignment/Acclimation 

Study  rats  were  randomized  on  19  January  1984  into  two 
experimental  dose  groups  of  68  males  and  68  females  each,  and 
a  saline  control  group  of  68  males  and  67  females. 

Allocation  was  accomplished  by  using  the  Beckman  TOXSYS™ 
Animal  Allocation  Program  (Beckman  Instruments,  Inc., 
Somerset,  NJ) .  The  animals  were  acclimated  for  12  days 
before  the  day  of  dosing.  During  this  period  they  were 
observed  daily  for  signs  of  illness. 

Dosage  Levels 

The  following  test  doses  were  administered:  high-dose 
group,  100  |Xl  IBC/animal;  low-dose  group,  10  p.1  IBC/animal; 
and  vehicle  control  group,  100  |Xl  saline/animal. 

Exspar  at  ion,  of  Compound 

Bucrylate®  IBC  tissue  adhesive  (lot  929-252,  Ethicon, 
Inc.,  Somerville,  NJ)  was  received  from  the  sponsor  on  10 
January  1984  as  750  0.5  ml  scored  ampules  in  individual 
overwrap  sterile  bags.  The  IBC  required  no  preparation  prior 
to  implantation.  It  was  stored  at  LAIR  at  room  temperature 
under  darkness.  Dosing  of  control  animals  was  performed  with 
commercial  sterile  isotonic  (0.9%)  saline  (Lot  No.  56-329-FD- 
05,  expiration  date  1  Sep  86,  Abbott  Labs,  Chicago,  IL) . 

Chemical  Analysis  of  IBC 

Ethicon,  Inc.,  provided  data  on  infrared, 
chromatographic,  and  chemical  analysis  of  the  IBC  (Appendix 
A).  Lot  929-252  contained  99.9%  (w/w)  total  monomer.  Before 
dosing,  the  LAIR  Analytical  Chemistry  Group  verified  the 
identity  of  the  IBC  by  IR  spectroscopy,  confirmed  the  purity 


! 


Brown  et  al . — 4 


by  gas  chromatography,  and  demonstrated  the  stability  of  the 
IBC  during  the  dosing  period  (Appendix  A) . 

Test  Procedures 

The  fixed  volume  of  neat  IBC  or  saline  each  animal 
received  was  based  upon  its  assigned  dosage  group,  and  was 
administered  directly  onto  the  ventral  (inferior  surface) 
capsule  of  the  liver.  High-dose  animals  received  100  ill  of 
IBC,  low-dose  animals  received  10  |ll  of  IBC,  and  control 
animals  received  100  ill  of  saline.  Rats  were  6  weeks  of  age 
at  dosing.  Since  the  surgical  survival  rate  for  implantation 
of  this  compound  was  unknown,  all  animals,  regardless  of 
size,  were  subjected  to  the  surgical  procedure  and  dosed  to 
ensure  that  sufficient  numbers  would  be  available  for 
inclusion  in  the  chronic  phase  of  the  study.  Body  weights  at 
dosing  ranged  from  29  g  to  123  g,  with  the  mean  male  and 
female  weights  at  94.0  g  and  73.8  g,  respectively. 

The  test  compound  was  implanted  under  sterile  conditions 
in  the  LAIR  Operating  Room  Suite  on  23  and  24  Jan  84.  The 
surgical  laparotomy  implantation  procedure  was  performed 
under  xylazine/ketamine  anesthesia.  Following  anesthesia, 
surgical  preparation,  and  midline  incision,  the  animal's 
liver  was  exposed  by  "tenting”  the  abdominal  wall  with  either 
ophthalmic  retractors  or  tissue  forceps.  The  IBC  was  applied 
to  the  ventral  capsule  of  the  liver,  usually  the  caudate 
lobe,  by  using  either  a  sterile  tip  100  Jll  or  10  }il  fixed 
volume  Eppendorf  micropipette.  The  IBC  was  then  allowed  to 
polymerize  for  2-3  minutes  before  closure.  All  pipettes  used 
in  the  study  were  shown  to  be  accurate  within  ±2%.  A  two- 
layer  closure  of  bioresorbable  4-0  Vicryl®  (Ethicon,  Inc.) 
and  skin  staples  or  skin  clips  was  used.  After  the  animals 
recovered  to  sternal  recumbency,  they  were  returned  to  the 
Toxicology  Suite.  A  complete  surgical  report  is  provided  in 
Appendix  C. 

Clinical  Observations 

On  the  day  of  dosing  and  during  the  following  2-week 
period,  animals  were  checked  intermittently  throughout  the 
day.  During  the  first  postoperative  week,  25  animals  were 
removed  from  the  study  due  to  underweight  condition  and/or 
complications.  Three  hundred  eighty-two  animals  recovered 
satisfactorily  and  remained  on  study  as  of  1  Feb  84.  After 
the  animals  were  stabilized,  observations  for  mortality  and 
signs  of  toxicity  or  illness  were  reduced  in  frequency  to 
twice  daily.  An  observation  was  performed  according  to  the 
following  procedure:  (a)  each  morning  all  animals  were 
observed  closely  for  signs  of  toxicity  or  illness  without 


Brown  et  al. 


disturbing  them  in  the  cage;  (b)  once  a  week  the  animals  (20% 
cf  them  per  work  day)  were  removed  from  their  cages  and 
observed;  (c)  animals  were  observed  after  being  returned  to 
their  cages.  A  second  "walk-through,  live/dead  check" 
observation  was  performed  every  afternoon  with  only 
significant  observations  recorded.  Monthly,  a  more  detailed 
clinical  examination  was  performed,  body  weights  were 
obtained,  and  the  abdomens  of  all  animals  were  gently 
palpated.  These  data  were  recorded  electronically  on  a 
Beckman  TOXSYS™  Data  Collection  Terminal.  Daily 
observat ions/clinical  signs  were  recorded  on  written  records 
and  significant  changes  entered  into  the  TOXSYS  workstation. 
TOXSYS  software  on  the  LAIR  Data  General  Computers,  Models 
MV8000  and  C330,  was  used  to  analyze  clinical  signs  and  body 
weight  data. 

Pathological  Examinations 

Animals  that  were  moribund  at  the  time  of  clinical 
examination  were  euthanized  by  pentobarbital  overdose, 
exsanguinated  by  axillary  incision,  and  necropsied.  During 
the  second  year,  21  animals  died  unexpectedly  (unscheduled) , 
and  97  unscheduled  sacrifices  were  required.  Additionally, 
187  animals  were  submitted  for  sacrifice  at  the  end  of  the 
second  year,  27  Jan  -  14  Feb  1986.  Gross  necropsy 
examinations  were  performed,  and  tissues  were  collected  from 
each  animal  in  accordance  with  LAIR  OP-STX-32,  "General 
Pathology  Procedures",  OP-PSG-7,  "Necropsy  Procedure — Gross 
Examination  of  Small  Laboratory  Animals",  and  OP-PSG-12, 
"Histopathology — Trimming  of  Rodent  Tissues." 

The  pathologic  evaluation  consisted  of  gross  and 
microscopic  examination  of  major  organs/tissues  and  all  gross 
lesions  from  sacrificed  animals  and  animals  found  dead.  The 
following  organs/tissues  were  examined  microscopically:  eyes 
and  lens,  skin,  subcutaneous  tissue,  mammary  gland,  brain  (4 
levels:  anterior  cerebral,  midcerebral,  midbrain, 
cerebellum) ,  middle  ears,  auditory  canal,  sebaceous  gland, 
trachea,  lungs,  nasal  region,  sternum,  heart,  aorta,  salivary 
glands  (parotid,  submaxillary,  sublingual) ,  Harderian 
lacrimal  and  intraorbital  lacrimal  glands,  exorbital  lacrimal 
glands,  liver  (2-4  sections  of  various  lobes),  pancreas, 
esophagus,  stomach,  duodenum,  jejunum,  ileum,  cecum,  colon, 
rectum,  kidneys,  urinary  bladder,  accessory  sex  glands  (male 
only — prostate,  seminal  vesicle,  coagulating  gland, 
epididymis),  testes/ovaries,  uterus  (horns  and  body), 
skeletal  muscle  (2  sections,  longitudinal  and  cross) ,  sciatic 
nerve,  tongue,  pituitary,  thyroid/parathyroid,  adrenals, 
thymus,  spleen  (2  cross  sections),  mesenteric  lymph  nodes, 
femur  (bone  marrow),  and  vertebrae  with  spinal  cord  (3 


Brown  et  al. — 6 


sections:  cervical,  thoracic,  and  lumbar).  These  tissues 
were  preserved  in  10%  buffered  formalin,  trimmed,  embedded  in 
paraffin,  sectioned,  and  stained  with  hematoxylin  and  eosin 
prior  to  microscopic  examination.  Necropsy  data  were 
recorded  and  entered  into  a  Xybion™  (Xybion  Medical  Systems, 
Cedar  Knoll,  NJ)  computerized  data  acquisition  program 
designed  for  a  DEC  VAX™  750  computer.  Microscopic  findings 
were  entered  into  the  computer  directly  as  microslides  were 
read.  Since  the  Xybion  animal-numbering  system  was 
incompatible  with  the  TOXSYS  numbering  system  used  during  the 
"in-life"  phases  of  the  study,  the  animals  had  to  be 
renumbered  by  the  Pathology  Section  as  they  were  necropsied 
in  order  to  enter  them  into  the  Xybion  Pathology  Data  System. 
A  cross  reference  of  animal  identification  numbers  is 
presented  in  Appendix  J. 

Statistical  Analyses 

Statistical  analyses  were  performed  on  the  study 
results.  TOXSYS  System  programs  were  used  to  determine  the 
group  mean  animal  body  weights  (EDS002)  and  the  frequencies 
of  clinical  signs  (EDS057-61) .  The  Xybion  Pathology  Data 
System  was  used  to  generate  the  pathology  raw  data  listings, 
summary  pathology  reports,  lesion  frequency  data,  and 
terminal  body  weights  (group  mean,  standard  deviation, 
Bartlett’s  Test,  ANOVA,  and  Dunnett's  Test).  The  incidence 
of  microscopic  lesions  for  each  test  group  were  compared  to 
the  control  group  using  the  Kolmogorov-Smirnov  two-tailed 
test.  The  5%  (p  ^  0.05)  level  of  significance  was  used  for 
all  tests . 

Duration  of  Study 

The  "in-life"  period  for  the  second  year  of  the  study 
ran  from  30  Jan  85  until  14  Feb  86,  when  the  terminal 
sacrifice  was  completed.  Appendix  D  is  a  complete  historical 
listing  of  study  events. 

Changes/Devlationa  from,  Protocol 

Performance  of  this  study  was  in  accordance  with  the 
protocol  and  applicable  amendments. 

Storage  of  Raw  Data  and  Final  Report 

A  copy  of  the  final  report,  study  protocol  and 
amendments,  raw  data,  relevant  SOPs,  analytical  data  for  the 
test  compound,  and  an  aliquot  of  the  test  compound  will  be 
retained  in  the  LAIR  Archives. 


Brown  et  al.--7 


RESULTS 

Mortality 

One  hundred  eighteen  animals  died  or  were  sacrificed 
prematurely  during  the  second  year  of  the  study  (controls,  13 
males,  22  females;  low  dose,  16  males,  19  females;  high  dose, 
27  males,  21  females) .  Of  these,  21  unscheduled  deaths  were 
recorded  and  97  animals  were  euthanized  because  they  were 
moribund,  had  infections,  had  lost  weight,  or  were  in  poor 
condition  (Tables  la  -  Id) . 

TABLE  la 

Listing  of  Unschadulsd  Dsaths/Euthanixsd*  Animals 

1  Fab  -  30  Apr  85 


Toxicology 

Dose 

Animal  No. 
84D00- 

Date 

Clinical  Outcome 

Sex 

Group 

376 

13 

Feb 

85 

Died  in  cage 

F 

High  IBC 

157 

26 

Feb 

85 

Died  in  cage 

M 

High  IBC 

296 

15 

Mar 

85 

Sacrificed,  large** 
intra-abdominal  mass 

F 

High  IBC 

170 

23 

Apr 

85 

Sacrificed,  alopecia  and 
skin  scabbing/scaling 
(infection  control) 

M 

Control 

428 

23 

Apr 

85 

Sacrificed,  perivaginal 
fistula  &  discharge 

F 

Control 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 

**Large  -  £  20  mm  in  diameter 


Brown  et  al. — 8 


TABLE  lb 


Listing  of  Unscheduled  Desths/Euthanized*  Animals 

1  May  -  31  Jul  85 


Toxicology 
Animal  No. 
84DQQ- _ 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

422 

7 

May 

85 

Sacrificed,  severe 
dehydration 

F 

Low  IBC 

156 

9 

May 

85 

Died  in  cage 

M 

High  IBC 

423 

10 

May 

85 

Sacrificed,  large** 
intra-abdominal  mass 

F 

High  IBC 

181 

10 

May 

85 

Sacrificed,  large 
intra-abdominal  mass 

M 

High  IBC 

109 

19 

May 

85 

Sacrificed,  large 
intra-abdominal  mass 

M 

High  IBC 

384 

31 

May 

85 

Sacrificed,  skin  lesions 
neck  (infection  control) 

F 

Control 

129 

5 

Jun 

85 

Large  intra-abdominal 

mass 

M 

High  IBC 

275 

14 

Jun 

85 

Large  intra-abdominal 

mass 

F 

High  IBC 

326 

1 

Jul 

85 

Died  in  cage 

F 

Low  IBC 

431 

8 

Jul 

85 

Large  intra-abdominal 

mass 

F 

High  IBC 

430 

9 

Jul 

85 

Died  in  cage 

F 

High  IBC 

415 

17 

Jul 

85 

Pallor  with  palpable 
splenic  mass 

F 

Low  IBC 

193 

21 

Jul 

85 

Died  in  cage 

M 

High  IBC 

091 

30 

Jul 

85 

Died  in  cage 

M 

High  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 


**Large  -  £  20  mm  in  diameter 


Brown  et  al. — 9 


TABLE  lc 


Listing  of  Unscheduled  Deaths/Euthanized*  Animals 

1  Aug  -  31  Oct  85 


Toxicology 
Animal  No. 
84D00- 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

099 

1 

Aug 

85 

Large**  intra-abdominal 
mass 

M 

High  IBC 

149 

1 

Aug 

85 

Emaciated  and  dehydrated 

M 

Low  IBC 

301 

1 

Aug 

85 

Malocclusion 

F 

Control 

191 

1 

Aug 

85 

Infected  subcutaneous 
mass 

M 

High  IBC 

286 

2 

Aug 

85 

Pallor  with  palpable 
abdominal  mass 

F 

High  IBC 

388 

2 

Aug 

85 

Pallor  with  palpable 
abdominal  mass, 
died  in  cage 

F 

Control 

147 

16 

Aug 

85 

Large  subcutaneous 
perihumeral  mass 

M 

Low  IBC 

407 

16 

Aug 

85 

Large  subcutaneous 
mass  on  thoracic  wall 

F 

Low  IBC 

279 

20 

Aug 

85 

Marked  weight  loss  and 
perianal  staining 

F 

Low  IBC 

081 

28 

Aug 

85 

Large  subcutaneous  mass 

M 

High  IBC 

426 

28 

Aug 

85 

Pallor  with  weight  loss 

F 

Low  IBC 

074 

30 

Aug 

85 

Large  intra-abdominal  mass 
with  excessive  weight  loss 

M 

High  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 


**Large  »  t  20  mm  in  diameter 


Brown  efc  al. — 10 


TABLE  lc  (COnt.) 

Listing  of  Unschsdulsd  Deaths/Euthanized*  Animals 

1  Aug  -  31  Oct  85 


Toxicology 

Dose 

Animal  No. 
84D00- 

Date 

Clinical  Outcome 

Sex 

Group 

102 

30 

Aug 

85 

Large**  intra-abdominal 
mass  and  pallor 

M 

Low  IBC 

108 

30 

Aug 

85 

Skin  mass  over  maxilla 

M 

High  IBC 

151 

30 

Aug 

85 

Pallor  with  large 
intra-abdominal  mass 

M 

High  IBC 

327 

30 

Aug 

85 

Large  intra-abdominal  mass 

F 

High  IBC 

338 

30 

Aug 

85 

Large  intra-abdominal  mass 

F 

High  IBC 

180 

9 

Sep 

85 

Large  intra-abdominal  mass 
with  weight  loss 

M 

Low  IBC 

288 

9 

Sep 

85 

Emaciated 

F 

Control 

363 

13 

Sep 

85 

Oral  discharge 

F 

Low  IBC 

389 

13 

Sep 

85 

Cardiac  murmur  with 
weight  loss 

F 

Control 

032 

19 

Sep 

85 

Large  subcutaneous  mass 

M 

Control 

404 

23 

Sep 

85 

Died  in  cage 

F 

Control 

260 

25 

Sep 

85 

Enteritis  with  weight  loss 

F 

Low  IBC 

059 

26 

Sep 

85 

Large  subcutaneous  mass 

M 

Low  IBC 

095 

26 

Sep 

85 

Large  intra-abdominal  mass 

M 

High  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 

**Large  =  £  20  mm  in  diameter 


Brown  et 


TABLE  lc  (cont.) 


Listing  of  Unscheduled  Deaths/Euthanised*  Animals 

1  Aug  -  31  Oct  85 


Toxicology 
Animal  No. 
84DQQ- _ 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

104 

26 

Sep 

85 

Large**  intra-abdominal 
mass 

M 

High  IBC 

124 

26 

Sep 

85 

Large  splenic  mass 

M 

Control 

266 

3 

Oct 

85 

Large  subcutaneous 
mass  on  neck 

F 

Control 

069 

3 

Oct 

85 

Large  intra-abdominal 
mass  with  weight  loss 

M 

High  IBC 

368 

8 

Oct 

85 

Severe  pallor  from 
hemorrhagic  enteritis 

F 

Control 

199 

16 

Oct 

85 

Deteriorated  condition 
and  large  palpable 
intra-abdominal  mass 

M 

High  IBC 

278 

17 

Oct 

85 

Large  subcutaneous 
mass  over  humerus 

Y 

Control 

166 

17 

Oct 

85 

Large  subcutaneous  mass 

M 

Control 

084 

24 

Oct 

85 

Pallor,  weak,  depressed 

M 

Low  IBC 

080 

24 

Oct 

85 

Died  in  cage 

M 

High  IBC 

203 

24 

Oct 

85 

Pallor,  large 
intra-abdominal  mass 

M 

Low  IBC 

424 

25 

Oct 

85 

Weight  loss,  dehydrated, 
depressed 

F 

High  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 

**Large  *  i  20  mm  in  diameter 


Brown  et  al. — 12 


TABLE  Id 


Listing  of  Unschsdulsd  Deaths/Euthanized*  Animals 

1  Nov  85  -  14  Fab  86 


Toxicology 
Animal  No. 
84DQQ- _ 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

139 

5 

Nov 

85 

Large**  intra-abdominal 
mass  with  weight  loss 

M 

High  IBC 

416 

5 

Nov 

85 

Large  intra-abdominal  mass 
with  deterioration 

F 

High  IBC 

164 

8 

Nov 

85 

Weight  loss 

M 

Control 

320 

15 

Nov 

85 

Large  intra-abdominal  mass, 
anorexic 

F 

High  IBC 

390 

15 

Nov 

85 

Anorexic,  pallor 

F 

Control 

146 

18 

Nov 

85 

Died  in  cage 

M 

Low  IBC 

343 

19 

Nov 

85 

Large  intra-abdominal  mass 

F 

High  IBC 

367 

19 

Nov 

85 

Large  mass  lower  abdomen 

F 

High  IBC 

414 

19 

Nov 

85 

Large  subcutaneous  mass 

F 

Control 

273 

18 

Nov 

85 

Weight  loss,  palpable 
intra-abdominal  mass 

F 

High  IBC 

322 

23 

Nov 

85 

Died  in  cage  after  monthly 
palpation,  large  splenic 
mass 

F 

Low  IBC 

365 

23 

Nov 

85 

Splenic  mass,  died  in  cage 

F 

Control 

182 

26 

Nov 

85 

Large  intra-abdominal  mass, 
pallor 

M 

Low  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 


**Large  -  £  20  mm  in  diameter 


Brown  et  al . --13 


TABLE  Id  (cont.) 

Listing  of  Unschsdulsd  Deaths/Euthanized*  Animals 

1  Mov  85  -  14  Fab  86 


Toxicology 
Animal  No. 
84D0fi- 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

290 

26 

Nov 

85 

Pallor 

F 

Control 

036 

3 

Dec 

85 

Anorexic,  weight  loss 

M 

Control 

334 

3 

Dec 

85 

Pallor  and  emaciated 

M 

Control 

392 

3 

Dec 

85 

Emaciated  and  pallor 

F 

Low  IBC 

041 

11 

Dec 

85 

Emaciated 

M 

Low  IBC 

062 

11 

Dec 

85 

Large**  intra-abdominal 
mass,  rapid  weight  loss 

M 

High  IBC 

030 

18 

Dec 

85 

Anorexic,  emaciated 

M 

Control 

033 

19 

Dec 

85 

Died  in  cage 

M 

Low  IBC 

070 

23 

Dec 

85 

Rapid  weight  loss 

M 

Low  IBC 

051 

23 

Dec 

85 

Weight  loss 

M 

Low  IBC 

359 

23 

Dec 

85 

Died  in  cage 

F 

Control 

200 

24 

Dec 

85 

Weight  loss,  circumoral 
staining 

M 

Control 

399 

29 

Dec 

85 

Anorexic,  emaciated,  skin 
pallor 

F 

Low  IBC 

078 

3 

Jan 

86 

Emaciated,  pallor,  large 
intra-abdominal  mass 

M 

High  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 


**Large  =  £  20  mm  in  diameter 


Brown  et  al . — 14 


TABLE  Id  (cont . ) 

Listing  of  Unscheduled  Deaths/Euthanized*  Animals 

1  Nov  85  -  14  Feb  86 


Toxicology 
Animal  No. 
84DQQ- _ 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

114 

3 

Jan 

86 

Pallor,  emaciated,  large** 
intra-abdominal  mass 

M 

High  IBC 

185 

3 

Jan 

86 

Large  intra-abdominal  mass, 
pallor 

M 

High  IBC 

328 

3 

Jan 

86 

Pallor  and  emaciated 

F 

Low  IBC 

060 

9 

Jan 

86 

Large  intra-abdominal  mass 

M 

Low  IBC 

289 

11 

Jan 

86 

Died  in  cage 

F 

Low  IBC 

257 

13 

Jan 

86 

Large  subcutaneous  tumor 

F 

Low  IBC 

324 

13 

Jan 

86 

Acute  weight  loss 

F 

Control 

345 

13 

Jan 

86 

Emaciated 

F 

Low  IBC 

089 

14 

Jan 

86 

Died  in  cage 

M 

High  IBC 

100 

14 

Jan 

86 

Convulsed  and  died  in  cage 

M 

High  IBC 

120 

14 

Jan 

86 

Moribund 

M 

Control 

117 

14 

Jan 

86 

Moribund 

M 

Control 

053 

16 

Jan 

86 

Two  large  intra-abdominal 
masses 

M 

Low  IBC 

039 

16 

Jan 

86 

Weight  loss 

M 

Control 

198 

16 

Jan 

86 

Large  intra-abdominal  mass 

M 

Control 

*  Animals  were  sacrificed  to  conserve  tissues  ir  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 

**La’-ge  =  £  20  mm  in  diameter 


Brown  et  al.  — 15 


TABLE  Id  (COnt  .  ) 


Listing  of  Unschsdulsd  Osaths/Euthanizsd*  Animals 

1  Nov  85  -  14  Fab  86 


Toxicology 

Dose 

Animal  No. 
84D00- 

Date 

Clinical  Outcome 

Sex 

Group 

413 

16 

Jan 

86 

Subcutaneous  swelling 
between  anus  and  vulva, 
intra-abdominal  mass 

F 

High  IBC 

280 

17 

Jan 

86 

Pallor,  enlarged  spleen 

F 

Control 

356 

17 

Jan 

86 

Malocclusion,  weight  loss, 
intra-abdominal  mass 

F 

High  IBC 

397 

17 

Jan 

86 

Pallor,  enlarged  spleen 

F 

Control 

425 

17 

Jan 

86 

Deteriorated  condition, 
hypothermic,  with  weight 
loss 

F 

Low  IBC 

418 

17 

Jan 

86 

Splenic  and  liver  masses, 
hypothermic 

F 

Low  IBC 

346 

21 

Jan 

86 

Moribund 

F 

Low  IBC 

412 

21 

Jan 

86 

Died  in  cage 

F 

High  IBC 

172 

22 

Jan 

86 

Died  in  cage 

M 

High  IBC 

393 

23 

Jan 

86 

Deteriorated  condition 

F 

High  IBC 

432 

23 

Jan 

86 

Died  in  cage 

F 

High  IBC 

115 

24 

Jan 

86 

Moribund 

M 

High  IBC 

096 

27 

Jan 

86 

Emaciated,  hypothermic, 
severe  respiratory  problem 

M 

Low  IBC 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 


**Large  =  £  20  mm  in  diameter 


Brown  et  al . — 16 


TABLE  Id  (cont.) 


Listing  of  Unscheduled  Deaths/Euthanized*  Animals 

1  Nov  85  -  14  Fab  86 


Toxicology 
Animal  No. 
84D0Q- 

Date 

Clinical  Outcome 

Sex 

Dose 

Group 

361 

29 

Jan 

86 

Emaciated,  intra-abdominal 
mass 

F 

Low  IBC 

179 

29 

Jan 

86 

Deteriorated  condition 

M 

Control 

302 

3 

Feb 

86 

Two  intra-abdominal  masses 
emaciated,  pallor 

,  F 

High  IBC 

330 

7 

Feb 

86 

Emaciated,  pallor, 
perianal  stain 

F 

Control 

331 

7 

Feb 

86 

Splenic  mass,  pallor 

F 

Control 

403 

10 

Feb 

86 

Emaciated,  sudden  onset 
conjunctivitis 

F 

Control 

*  Animals  were  sacrificed  to  conserve  tissues  in  face  of 
imminent  death,  for  infection  control  purposes,  or  for 
humane  reasons . 

Clinical  Observations 

A  glossary  of  terms  for  clinical  observations  is 
presented  in  Appendix  E.  Individual  clinical  observation  and 
body  weight  data  are  presented  in  Appendix  F.  The  majority 
of  animals  gained  or  maintained  body  weight  throughout  the 
first  8  months  of  the  second  year  of  the  study  (observation 
periods  14-23) .  During  the  final  4  months  (observation 
periods  24-27),  slight  decreases  in  group  mean  body  weights 
were  observed  for  treatment  and  control  group  males. 

Decreases  were  observed  for  the  females  only  at  terminal 
sacrifice  (fasted  body  weight) .  Mean  body  weights  of  IBC- 
dosed  and  saline-treated  control  animals  were  comparable 
throughout  the  second  year  for  both  sexes.  Tables  2  (male) 
and  3  (female)  present  mean  monthly  body  weights  by  group. 

Clinical  signs  closely  duplicating  those  seen  during  the 
first  year  of  the  study  (1)  were  observed  throughout  the 
second  year.  With  the  exception  of  clinically  observed 


Brown  et  al.  — 17 


masses,  signs  were  generally  routine,  of  minor  consequence, 
and  were  distributed  relatively  equally  among  the  treatment 
and  control  groups.  Clinical  sign  categories  observed  during 
the  second  year  included  ocular  abnormalities  (179  of  305 
animals  affected),  masses  (145  of  305),  skin/fur 
abnormalities  (129  of  305),  general  signs  (69  of  305), 
respiratory  abnormalities  (23  of  305) ,  genitourinary 
abnormalities  (14  of  305),  behavioral  aberrations  (12  of 
305),  postoperative  complications  (5  of  305),  and 
miscellaneous  observations  (21  ear  tag  reactions,  2 
malocclusions,  1  cardiac  irregularity) . 

With  the  exceptions  of  the  masses,  ocular  abnormalities, 
and  the  few  iatrogenic  postoperative  complications  (xyphoid 
protuberance  and  abdominal/peritoneal  fistula) ,  clinical 
signs  developed  rapidly,  and  were  relatively  mild  and  of 
short  duration.  An  incidence  summary  of  clinical  signs  is 
presented  in  Table  4 . 


TABLE  2 

Group  Moan  Body  Weights :  Malta  - 

Yaar  Two  of  Two-Year  IBC  Carcinogenicity  Bioassay* 


Date  Observation  High  Dose  Low  Dose 

Period  IBC  IBC 


Saline 

Controls 


14 

Feb 

85 

14 

396.5 

(50) 

400.9 

(55) 

404.2 

(57) 

14 

Mar 

85 

15 

404.2 

(49) 

407.8 

(55) 

410.3 

(57) 

11 

Apr 

85 

16 

407.5 

(49) 

409.5 

(55) 

414.8 

(57) 

9 

May 

85 

17 

410.7 

(49) 

415.7 

(55) 

420.9 

(56) 

6 

Jun 

85 

18 

417.8 

(45) 

419.6 

(55) 

422.7 

(56) 

2 

Jul 

85 

19 

416.2 

(45) 

419.4 

(55) 

422.3 

(56) 

1 

Aug 

85 

20 

420.6 

(43) 

421.8 

(55) 

425.8 

(56) 

29 

Aug 

85 

21 

415.7 

(40) 

422.4 

(53) 

42  7.3 

(56) 

26 

Sep 

85 

22 

417.8 

(37) 

423.5 

(51) 

426.1 

(55) 

24 

Oct 

85 

23 

417.8 

(33) 

425.5 

(50) 

431.5 

(53) 

21 

Nov 

85 

24 

407.5 

(31) 

415.6 

(47) 

420.3 

(52) 

19 

Dec 

85 

25 

406.3 

(30) 

412.8 

(45) 

416.7 

(50) 

16 

Jan 

86 

26 

401.4 

(25) 

403.8 

(41) 

404.4 

(47) 

27  Jan  - 
5  Feb  86** 

27 

376.2 

(23) 

377.5 

(39) 

376.8 

(44) 

*  Data  are  presented  as  group  mean  value  in  grams,  with  n, 
the  number  of  animals  in  each  group,  in  parentheses. 

**  Final  sacrifice  period,  fasted  body  weights 


Brown  et  al . — 18 


TABLE  3 

Group  Moan  Body  Haights :  Famalas  - 

Year  Two  of  Two-Yaar  IBC  Carcinoganicity  Bioassay* 


Date 

Observation  High  Dose 

Period  IBC 

Low  Dose 

IBC 

Saline 

Controls 

15 

Feb 

85 

14 

232.7 

(40) 

227.8 

(52) 

225.3 

(51) 

15 

Mar 

85 

15 

241.0 

(40) 

233.2 

(52) 

231.9 

(51) 

12 

Apr 

85 

16 

248.7 

(39) 

240.3 

(52) 

236.4 

(51) 

10 

May 

85 

17 

255.4 

(39) 

245.2 

(51) 

244.8 

(50) 

7 

Jun 

85 

18 

261.2 

(38) 

252.1 

(51) 

246.8 

(49) 

3 

Jul 

85 

19 

263.8 

(37) 

256.8 

(50) 

253.1 

(49) 

2 

Aug 

85 

20 

270.1 

(34) 

264.1 

(49) 

261.1 

(48) 

30 

Aug 

85 

21 

274.6 

(33) 

265.8 

(46) 

263.6 

(47) 

27 

Sep 

85 

22 

281.2 

(31) 

273.1 

(44) 

273.2 

(44) 

25 

Oct 

85 

23 

281.2 

(31) 

276.5 

(44) 

273.8 

(41) 

22 

Nov 

85 

24 

281.4 

(26) 

278.9 

(44) 

274.1 

(39) 

20 

Dec 

85 

25 

281.6 

(25) 

278.2 

(42) 

276.5 

(36) 

17 

Jan 

86 

26 

279.1 

(25) 

278.0 

(37) 

274.1 

(34) 

7- 

14  Feb 

86**27 

261.2 

(19) 

264.6 

(33) 

256.7 

(29) 

*  Data  are  presented  as  group  mean  value  in  grams,  with  n, 
the  number  of  animals  in  each  group,  in  parentheses. 

**  Final  sacrifice  period,  fasted  body  weights 

The  clinically  observed  masses  included  intra-abdominal 
masses,  subcutaneous  masses,  inguinal  masses,  cutaneous 
masses  of  the  ear  or  scrotum,  a  maxillary  mass,  a  subscapular 
mass,  a  swollen  maxilla,  and  splenomegaly.  Of  these,  only 
the  intra-abdominal  group  could  be  correlated  to  IBC- 
treatment.  The  intra-abdominal  masses  were  primarily 
associated  with  the  ventral  aspect  of  the  liver.  An 
increased  incidence  of  intra-abdominal  masses  was  observed 
for  the  males  relative  to  the  females,  and  a  dose  response 
was  apparent  for  both  sexes . 

The  ocular  signs  (conjunctivitis,  corneal  opacity, 
opaque  lens,  prolapsed  lens,  hyphema,  ocular  discharge/ 
chromodacryorrhea,  microphthalmia,  exophthalmus/enlarged 
globe,  iris  tear,  and  dilated  pupil)  observed  in  many  of  the 
animals  (179  of  305)  were  sequelae  to  drying 
(keratoconjunctivitis  sicca)  of  the  eyes  during  surgery 
(Appendix  C) .  Corneal  ulcers  formed  within  7-14  days.  These 
lesions  healed,  leaving  persistent  corneal  scars. 


Brown  et  al . — 19 


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Incidence  Summary  of  Clinical  Signs 


Brown  et  al . — 20 


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Brown  et  al . — 21 


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Brown  et  al . — 23 


Some  animals  also  had  conjunctivitis  and 
chromodacryorrhea  which  persisted  for  long  periods. 
Streptococcus  faecalis,  a  common  normal  flora  bacterium,  was 
cultured  from  the  affected  eyes.  The  consulting 
ophthalmologist  reported  that  this  low-grade  ocular  infection 
was  limited  to  the  globe  and  its  accessory  structures. 
Therefore,  no  treatment  was  rendered.  Other  postsurgical 
complications  included  xyphoid  cartilage  protuberance/nodule 
(due  to  accidental  incision  of  the  xyphoid  cartilage  in  some 
animals  while  attempting  to  expose  the  liver  sufficiently) 
and  peritoneal  fistula.  Significant  postoperative 
complications  occurred  in  less  than  5%  of  all  animals. 

Gross  Pathology 

Incidence  summary  reports  for  gross  necropsy 
observations  during  year  two  are  presented  in  Appendix  G. 

Twenty-one  animals,  10  males  and  11  females,  were  found 
dead,  and  97  animals  (46  males  and  51  females)  were 
euthanized  prematurely  during  the  second  year  of  the  study. 
Compound- related  lesions  (liver  adhesions)  were  observed  in 
70  of  83  IBC-treated  animals  in  the  unscheduled  group.  One 
hundred  eighty-seven  animals  (106  males  and  81  females) 
survived  to  the  end  of  the  study  and  were  euthanized. 
Compound-related  lesions  (liver  adhesions)  were  observed  in 
111  of  114  IBC-treated  animals. 

Dense,  tough,  fibrous  adhesions  were  observed  on  the 
capsular  surface  of  the  liver  in  48/50  Group  1  (100  p.1  IBC) 
males,  34/40  Group  1  (100  )ll  IBC)  females,  52/55  Group  2  (10 
H.1  IBC)  males,  and  47/52  Group  2  (10  |ll  IBC)  females.  The 
microscopic  characteristics  of  this  test  compound- related 
inflammatory  reaction  are  described  in  detail  below  in  the 
paragraphs  dealing  with  histologic  findings.  Only  1/57  Group 
3  (saline  control)  males  and  3/51  Group  3  females  had  grossly 
evident  adhesions  on  the  liver.  The  adhesions  in  control 
animals  were  very  fine,  delicate  strands  in  marked  contrast 
to  the  extensive,  tough,  fibrous  areas  observed  in  IBC- 
treated  animals . 

Liver  masses  were  observed  grossly  in  9/50  Group  1  (100 
H.1  IBC)  males,  13/40  Group  1  (100  fil  IBC)  females,  3/55  Group 
2  (10  p.1  IBC)  males,  and  0/53  Group  2  (10  )ll  IBC)  females. 
Grossly,  these  irregularly-shaped  masses  were  usually  white 
to  light  yellow/gray  in  color,  and  very  firm.  In  several 
masses  there  was  gross  evidence  of  hemorrhage  and/or 
necrosis.  Gross  observation  revealed  no  liver  masses  in  any 
Group  3  (saline  control)  male  or  female  rats.  In  some  of  the 
IBC-treated  animals,  masses  were  noted  in  organs  other  than 


Brown  et  al . — 24 


the  liver,  including  the  diaphragm,  stomach,  intestines, 
other  abdominal  organs,  and  in  the  thoracic  cavity.  Gross 
appearance  of  these  extrahepatic  masses  was  similar  to  that 
of  the  liver  masses.  Histologic  examination  revealed  that 
masses  in  the  liver  and  other  sites  were  sarcomas  arising  in 
the  compound-related  chronic  inflammatory  reaction.  The 
incidence  of  these  tumors,  a  detailed  histologic  description, 
and  anatomic  distribution  is  provided  in  the  paragraphs  below 
dealing  with  histologic  findings . 

Microscopic  Pathology 

In  the  text  of  this  report,  the  description  of 
histologic  lesions  is  limited  to  those  lesions  that  can  be 
attributed  to  administration  of  the  test  compound,  and 
lesions  or  disease  entities  that  may  affect  results  or 
interpretation  of  results.  A  complete  listing  of  all 
microscopic  findings  (neoplastic  and  non-neoplastic)  in  this 
study  is  provided  in  Appendix  H.  Appendix  I  is  a  glossary  of 
pathologic  terms  used  in  this  study.  It  includes  a 
definition,  a  brief  morphologic  description,  and  criteria  for 
severity  grades  of  all  microscopic  lesions  observed  in  this 
study . 

Non-Neoplastic  Lesions: 

An  incidence  summary  for  all  non-neoplastic  lesions  in 
this  study  is  presented  in  Appendix  H.  Non-neoplastic 
lesions  of  statistically  significant  (p  £  0.05)  incidence 
include  compound-related  inflammatory  reactions  in  the  liver 
and  different  locations  of  the  gastrointestinal  tract  in  both 
males  and  females  receiving  the  high  dose  of  IBC,  hepatocytic 
vacuolization  in  high-dose  females,  the  presence  of  red  blood 
cells  and/or  hemosiderin  within  lymph  nodes  of  high-dose 
males  and  females,  and  reactive  hyperplasia  of  lymph  nodes  in 
males  and  females  in  both  high  and  low  dose  groups . 

Compound-related  inflammatory  reactions  were  present  in 
nearly  all  animals  in  both  high  and  low  dose  IBC  groups. 

Table  5  presents  the  incidence  and  distribution  of  these 
lesions  in  abdominal  viscera.  In  most  animals,  the  lesions 
were  located  at  the  site  where  the  test  material  was  placed, 
i.e.,  on  the  capsule  of  the  liver  and  the  adjacent  diaphragm. 
Fewer  lesions  were  present  on  surfaces  of  the  stomach, 
different  portions  of  the  intestinal  tract,  spleen,  and 
pancreas.  The  microscopic  characteristics  of  compound- 
related  inflammatory  reactions  were  similar  in  all  sites.  In 
histologic  sections,  the  test  material  was  a  clear,  slightly 
yellow,  homogeneous  crystalline  material.  Surrounding  the 
test  material  there  was  a  granulomatous  foreign-body  reaction 


Brown  et  al  . — 25 


characterized  by  infiltration  of  macrophages,  lymphocytes, 
and  large  multinucleated  foreign-body  giant  cells  within 
abundant  fibrous  connective  tissue.  Usually  this  lesion  was 
limited  to  the  serosal  surface  of  abdominal  viscera.  In  some 
cases,  there  was  necrosis  of  underlying  tissues  and 
penetration  of  the  serosal  surface  by  the  inflammatory 
response  accompanied  by  regenerative  hyperplasia  of  cells  of 
the  organ  involved.  In  four  rats,  the  presence  of  the 
compound  caused  necrosis  of  the  intestinal  wall. 

Regenerating  cells  of  the  intestinal  mucosa  migrated  through 
the  submucosa  ani  muscular  tunics  of  the  intestine  and  were 
entrapped  in  the  adjacent  fibrous,  granulomatous  inflammatory 


TABLE  5 

Distribution  of  Compound-Related 
Inflammatory  Reactions:  Scheduled  and  Unscheduled 

Animals  Combined,  Year  2 


Group 

(Number  in  Group) 

CTLS* 

(57) 

MALE 

It 

(50) 

2§ 

(55) 

CTLS* 

(51) 

FEMALE 

It 

(40) 

2§ 

(52) 

Liver 

0 

48 

52 

0 

40 

45 

Diaphragm 

0 

41 

19 

0 

38 

25 

Jejunum 

0 

14 

0 

0 

13 

0 

Stomach 

0 

6 

0 

0 

12 

0 

Duodenum 

0 

7 

0 

0 

6 

0 

Ileum 

0 

7 

0 

0 

4 

0 

Pancreas 

0 

3 

0 

0 

1 

0 

Cecum 

0 

4 

0 

0 

1 

0 

Colon 

0 

3 

0 

0 

2 

0 

Spleen 

0 

1 

0 

0 

1 

0 

Abdominal  Cavity 

0 

1 

3 

0 

0 

0 

*  CTLS  =  Saline  controls 
t  Group  1  =  100  *ll  IBC 
§  Group  2  3  10  |ll  IBC 


Brown  et  al. — 26 


tissues.  In  these  cases,  the  entrapped  cells  had  essentially 
normal  histologic  characteristics,  except  for  their  location 
and  their  association  with  mild  degenerative  and  atrophic 
changes.  There  were  no  indications  of  cellular  atypia, 
nuclear  pleomorphism,  or  abnormal  mitotic  activity.  These 
lesions  were  present  in  the  jejunum  in  two  females  and  one 
male  in  the  high  dose  group,  and  in  the  duododenum  in  one 
high-dose  female.  No  intestinal  lesions  of  this  type  were 
observed  in  control  animals. 

The  incidence  of  hepatocytic  vacuolization  was 
statistically  significant  only  in  female  rats  in  the  high 
dose  group.  Seventeen  of  40  animals  in  this  group  exhibited 
the  lesion,  as  compared  to  8  of  51  control  females.  In  other 
treatment  groups,  the  lesion  was  present,  but  the  incidence 
was  not  statistically  significant.  Hepatocytic  vacuolization 
consisted  of  small-to-large  cytoplasmic  vacuoles  within 
hepatocytes.  In  sections  appropriately  prepared  and  stained, 
fat  could  be  demonstrated.  The  vacuolated  hepatocytes  were 
not  uniformly  distributed  in  the  liver. 

The  incidence  of  reactive  hyperplasia  in  lymph  nodes  was 
significantly  greater  in  male  and  female  rats  in  both  high 
and  low  dose  groups  compared  to  control  animals.  Reactive 
hyperplasia  consisted  of  non-neoplastic,  noninflammatory 
increases  in  cellularity  in  lymph  nodes.  There  were 
increased  numbers  of  cells  in  perifollicular  areas  of  the 
cortex.  Medullary  cords  and  sinuses  contained  large  numbers 
of  plasma  cells.  Eosinophilic,  protein-rich  edema  fluid  may 
have  been  present . 

The  presence  of  red  blood  cells  and/or  hemosiderin 
pigment  in  lymph  nodes  occurred  with  increased  frequency  in 
both  males  and  females  in  the  high  dose  group.  Table  6  shows 
the  incidence  or  these  changes  in  lymph  nodes. 

Neoplastic  Lesions: 

An  incidence  summary  for  all  neoplasms,  both  malignant 
and  benign,  is  presented  in  Appendix  H.  Sarcomas  that  arose 
in  the  compound-related  inflammatory  reaction  were  the  only 
neoplasms  that  exhibited  a  statistically  significant 
incidence.  Thirty-one  rats  treated  with  IBC  developed  these 
tumors.  Table  7  displays  the  distribution  of  these  lesions 
by  treatment  group,  sex,  and  whether  in  scheduled  or 
unscheduled  animals.  The  first  animal  necropsied  with  a 
compound- related  sarcoma  died  on  26  February  1985,  day  399 
after  intra-abdominal  administration  of  the  test  material. 


Brown  et  al. --27 


TABLE  6 

Incidence  of  Reactive  Hyperplasia, 
Red  Blood  Cells  and/or  Hemosiderin 
in  Lymph  Nodes,  Year  2 


MALE  FEMALE 


Group 

CTLS* 

It 

2§ 

CTLS* 

It 

2§ 

(Number  in  Group) 

(57) 

(50) 

(55) 

(51) 

(40) 

(52) 

Reactive  Hyperplasia 

8 

25** 

29** 

7 

25** 

24** 

RBC ' s  and/or  Hemosiderin 

12 

24** 

18 

15 

28** 

17 

*  CTLS  =  Saline  controls 
t  Group  1  =  100  Hi  IBC 
§  Group  2  =  10  Hi  IBC 

**Entries  flagged  with  (**'  are  significantly  different  from 
control  at  the  0.05  significance  level  using  Kolmogorov- 
Smirnov  Two-Tailed  Test. 


Of  the  31  animals  with  compound- related  sarcomas,  21  died  or 
were  sacrificed  in  a  moribund  condition  prior  to  scheduled 
termination  of  the  study.  Ten  sarcoma-affected  animals 
survived  until  terminal  sacrifice.  Only  27  of  the  31  animals 
with  sarcomas  had  grossly  detectable  masses  at  necropsy. 

Four  sarcomas  were  detected  only  by  microscopic  examination. 

These  sarcomas  involved  many  organs  in  some  animals, 
through  either  multicentric  sites  of  origin,  or  local 
invasion,  or  metastasis.  The  histologic  characteristics  of 
these  tumors  were  extremely  variable.  The  entire  spectrum  of 
histologic  differentiation  of  mesenchymal  tissue  was 
represented  in  this  group  of  tumors.  Some  tumors  were  well- 
differentiated  fibrosarcomas,  osteosarcomas,  or  myxosarcomas, 
while  others  were  very  anaplastic  and  poorly  differentiated. 
In  some  lesions,  multiple  types  of  histologic  differentiation 
occurred  in  the  same  tumor.  Of  the  31  compound-related 
sarcomas  in  this  study,  13  were  predominantly  fibrosarcomas, 
12  were  anaplastic  sarcomas,  5  were  osteosarcomas,  and  one 
was  a  myxosarcoma.  One  anaplastic  sarcoma  had  an  abundance 
of  chondrocytes  and  cartilaginous  matrix.  Two  other  tumors 


I 


Brown  et  al. — 28 


TABLE  7 

Sarcomas  Arising  in  Compound-Ralatad 
Inflammatory  Reaction 

Tima  of  Sacrifice:  Unscheduled  or  Terminal,  Year  2 


Group 

CTLS* 

MALE 

It 

2§ 

CTLS* 

female 

It 

2§ 

Total 

Unscheduled 

0 

9 

3 

0 

9 

0 

21 

Terminal** 

0 

6 

1 

0 

3 

0 

10 

Total 

0 

15 

4 

0 

12 

0 

31 

*  CTLS  -  Saline  controls 
t  Group  1  =  100  Hi  IBC 
§  Group  2  =  10  Hi  IBC 

**Terminal  or  final  sacrifice  (scheduled  animals) 


had  large  areas  that  resembled  rhabdomyosarcoma,  but  no  cross 
striations  could  be  demonstrated  in  sections  stained  with 
phosphotungstic  acid-hematoxylii:  stains.  The  liver  was  the 
most  frequently  affected  organ,  but  it  was  not  involved  in 
every  case.  Twenty-seven  of  the  31  animals  with  these  tumors 
had  lesions  in  the  liver.  In  one  rat,  only  the  duodenum  was 
involved,  one  animal  only  had  a  tumor  in  the  ileum,  and  in 
another,  the  sarcoma  was  limiued  to  the  stomach  and  spleen. 
Table  8  lists  the  anatomic  distribution  of  these  compound- 
related  sarcomas.  Because  of  the  multiple  sites  of 
involvement,  the  primary  site  for  these  lesions  is  coded  as 
the  abdominal  cavity,  and  involvement  of  each  individual 
organ  is  coded  as  a  metastatic  lesion.  Using  this  method  of 
computer  entry,  it  was  possible  to  tabulate  the  occurrence  of 
these  sarcomas  and  document  the  organs  involved  without 
duplicating  incidence.  The  diagnosis:  "Presence  of 
Metastatic  Sarcoma"  is  used  only  when  it  is  certain  that  the 
sarcoma  developed  within  the  reaction  to  the  test  material. 


TABLE  8 


Anatomic  Distribution  of  Sarcomas  Arising  in 
Compound-Related  Inflammatory  Reaction**,  Year  2 


Tissue 

CTLS* 

MALE 

It 

2§ 

CTLS* 

female 

It 

1 

Total 

Liver 

0 

12 

3 

0 

12 

0 

2  7 

Diaphragm 

0 

8 

2 

0 

10 

0 

i  9 

Mediastinal 

0 

4 

0 

0 

2 

0 

6 

Jejunum 

0 

2 

2 

0 

2 

0 

6 

Spleen 

0 

2 

2 

0 

1 

0 

5 

Pancreas 

0 

3 

1 

0 

1 

0 

5 

Stomach 

0 

1 

1 

0 

2 

0 

4 

Duodenum 

0 

3 

0 

0 

1 

0 

4 

Ileum 

0 

3 

0 

1 

0 

4 

Lung 

0 

1 

0 

/■N 

1 

0 

2 

Lymph  Nodes 

0 

0 

0 

0 

2 

0 

2 

Kidney 

0 

0 

0 

0 

1 

0 

1 

Cecum 

0 

1 

0 

0 

0 

0 

1 

Colon 

0 

1 

0 

0 

0 

0 

1 

*  CTLS  =  Saline  controls 
t  Group  1  =  100  n.1  IBC 
§  Group  2  =  10  |ll  IBC 

**Includes  both  primary  and  metastatic  sites 


Six  rats  in  this  study  had  fibrosarcomas  that  did  not 
arise  in  the  compound-related  inflammatory  reaction.  Four  of 
these  were  female  (1  control,  1  high  dose,  and  2  low  dose) 
animals  in  which  sarcomas  occurred  in  the  subcutaneous  tissue 
of  the  inguinal  region.  In  three  of  these  animals,  the 
tumors  invaded  the  adjacent  pelvic  cavity  and  involved  the 


Brown  et  al. --30 


rectum.  One  male  control  rat  had  a  fibrosarcoma  that  was 
limited  to  the  cortex  of  one  kidney.  One  male  in  the  high 
dose  group  had  a  fibrosarcoma  surrounding  the  metal 
identification  tag  in  the  pinna  of  the  external  ear.  Three 
females  (2  control  and  1  high  dose)  had  endometrial  stromal 
sarcomas.  These  lesions  did  not  resemble  the  sarcomas  that 
arose  in  compound-related  inflammatory  reaction. 

Four  IBC-treated  rats  (2  low  dose  males  and  2  high  dose 
females)  had  hepatocellular  carcinomas  in  the  liver.  The 
morphologic  characteristics  of  hepatocellular  carcinomas  are 
well  documented  in  the  literature.  For  this  study,  the 
criteria  of  Maronpot  et  al.  (6)  were  followed.  Important 
features  of  these  tumors  included  irregular  shape,  cellular 
atypia,  local  invasiveness,  haphazardly  arranged  cells,  broad 
sheets  of  cells,  trabecular  patterns,  and  glandlike 
formations  of  tumor  cells.  Hemorrhage,  necrosis,  vascular 
invasion,  and  metastases  were  observed  in  several  of  these 
lesions . 

Large  granular  lymphocyte  leukemia  occurred  in  62 
animals  in  this  study.  The  group  distribution  of  this 
malignant  tumor  of  lymphoid  cells  is  shown  in  Table  9.  The 
histologic  characteristics  of  this  common  neoplasm  in 
Fischer-344  rats  are  described  in  the  Glossary  of  Pathologic 
Terms,  Appendix  I. 


TABLE  9 

Large  Granular  Lymphocyte  Leukemia 
Time  of  Sacrifice:  Unscheduled  or  Terminal,  Year  2 


Group 

CTLS* 

MALE 

It 

2§ 

CTLS* 

FEMALE 

It 

2§ 

Total 

Unscheduled 

3 

9 

9 

9 

3 

9 

42 

Terminal** 

2 

1 

7 

5 

3 

2 

20 

Total 

5 

10 

16 

14 

6 

11 

62 

*  CTLS  =  Saline  controls 
t  Group  1  =  100  Hi  IBC 
§  Group  2  =  10  (11  IBC 

**Terminal  sacrifice  or  scheduled  animals 


There  was  no  statistical  significance  for  the  i:  Lder.ce  tf 
this  lesion;  however,  42  animals  died  or  were  sacrificed  ir.  c 
moribund  condition  because  of  the  presence  of  this  disease. 

Many  animals  in  this  study  developed  a  variety  of 
malignant  and  benign  tumors.  Tumors  of  the  testes,  endocrine 
glands,  and  mammary  glands  were  the  most  common.  There  was 
no  statistically  significant  increase  in  incidence  for  any  of 
these  miscellaneous  tumors  in  I3C-treated  animals  compared  tc 
controls . 

The  individual  animal  pathology  reports  are  provided  in 
Volume  2,  Appendix  J. 


DISCUSSION 

This  report  records  the  findings  from  the  second  year  of 
a  two-year  carcinogenicity  bioassay  of  IBC .  This  includes 
complete  clinical  histories  and  gross  and  microscopic 
pathological  data  on  all  unscheduled  and  scheduled  (final 
sacrifice)  animals  surviving  the  first  year  of  the  study. 

Clinical  findings  during  the  second  year  indicated  that 
male  and  female  animals  treated  with  IBC  gained  weight  at  the 
same  rate  as  the  control  animals.  These  findings  also 
indicated  that,  as  a  whole,  the  animals  in  this  study 
remained  relatively  healthy  throughout  the  second  year. 
Clinical  signs  observed  during  this  period  were  a 
continuation  of  those  seen  during  the  first  year  (1),  and 
generally  were  of  little  clinical  significance.  Ocular 
conditions,  which  were  secondary  to  desiccation  of  the  eyes 
during  surgery,  were  limited  to  the  globe  and  accessory 
structures,  and  had  no  significant  impact  on  the  animals' 
health.  Other  postsurgical  complications,  e.g.,  xyphoid 
protuberance,  were  also  insignificant  to  the  outcome  of  the 
study . 

One  hundred  eighteen  unscheduled  animals  died  or  were 
euthanized  at  various  times  during  the  second  year.  These 
animals  exhibited  more  aging  and  compound-related 
complications  than  did  animals  in  the  first  year.  IBC- 
treated  animals  in  the  unscheduled  group  had  compound-related 
gross  fibrous  adhesions  between  the  liver  and  the  diaphragm, 
abdominal  wall,  and/or  visceral  organs.  A  similar  lesion  was 
observed  in  IBC-treated  animals  euthanized  at  the  final 
sacrifice  (scheduled  animals) .  Microscopically,  these 
lesions  were  characterized  as  a  foreign-body  granulomatous 
reaction  involving  the  liver  capsule  and  parenchyma. 


Brown  et  al. — 32 


Analysis  of  the  results  of  this  study  indicates  that  the 
intra-abdominal  administration  of  IBC  is  related  to  the 
development  of  sarcomas  in  Fischer-344  rats.  Thirty-one 
(16%)  of  the  IBC-treated  animals  and  none  of  the  controls 
developed  these  tumors.  The  major  concern  is  the  mechanism 
by  which  the  test  material  exerts  its  carcinogenic  effect. 

The  results  of  this  study  do  not  indicate  definitively 
whether  chemical  carcinogenesis  or  solid-state  carcinogenic 
mechanisms  were  involved  in  the  induction  of  these  tumors. 

Solid-state  carcinogenesis  is  a  phenomenon  which  has 
been  recognized  for  nearly  fifty  years  (7) .  There  is  a  large 
body  of  literature  documenting  the  occurrence  of  this 
phenomenon,  and  comprehensive  reviews  of  this  topic  have  been 
published  (8,9,10).  Solid-state  carcinogenesis  involves  the 
development  of  sarcomas  in  the  subcutis  or  abdominal  cavity 
of  laboratory  rodents  that  have  had  solid,  smooth-surfaced 
material  implanted  in  these  sites.  A  wide  variety  of 
materials,  including  numerous  plastics,  cellophane,  teflon, 
metals,  and  glass  have  been  shown  to  induce  sarcomas  by  this 
mechanism.  The  chemical  nature  of  the  implanted  material  is 
irrelevant  to  the  production  of  tumors.  The  characteristics 
essential  for  tumor  production  are  the  size  and  form  of  the 
implant.  A  smooth  surface  is  essential.  Solid-state 
carcinogenesis  occurs  only  when  the  implanted  material  is  a 
film,  sheet,  or  other  large  form  having  a  smooth  surface.  If 
the  same  material  is  pulverized  and  administered  as  a  powder, 
no  tumors  result.  The  tumors  develop  in  the  fibrous, 
granulomatous,  foreign-body  reaction  that  envelopes  the 
implanted  material.  The  tumors  that  result  are  invariably 
sarcomas  that  have  diverse  histologic  variation. 

The  sarcomas  that  were  present  in  rats  implanted  with 
IBC  in  this  study  probably  developed  as  a  result  of  solid- 
state  carcinogenesis.  The  characteristics  of  the  implanted 
material,  the  anatomic  sites  involved,  and  the  histologic 
differentiation  of  the  sarcomas  fulfill  the  criteria  for  this 
mechanism.  However,  since  a  group  of  animals  implanted  with 
pulverized  IBC  was  not  included  in  the  experimental  design, 
chemical  carcinogenic  mechanisms,  though  an  unlikely  cause  of 
the  sarcomas,  cannot  be  completely  ruled  out  on  the  basis  of 
these  results. 

Six  animals  in  this  study  had  sarcomas  that  were  clearly 
unrelated  to  the  test  material.  Four  were  invasive 
fibrosarcomas  observed  in  the  subcutis  of  the  inguinal  region 
in  female  rats.  The  inguinal  fibrosarcomas  probably  resulted 
from  malignant  transformation  of  the  connective  tissue  stroma 
of  mammary  glands.  The  incidence  of  these  tumors  does  not 
differ  significantly  from  that  in  published  reports.  The 
fibrosarcoma  that  was  present  in  the  external  ear  adjacent  to 


Brown  et  al . — 33 


the  metal  identification  tag  in  one  animal  is  interesting  in 
that  it  may  represent  an  additional  case  of  solid-state 
carcinogenesis  in  response  to  a  foreign  body.  The  presence 
of  one  sarcoma  in  the  kidney  of  a  control  rat  is  interpreted 
as  a  spontaneous  lesion  unrelated  to  the  experiment,  and  of 
little  consequence. 

Four  IBC-treated  rats  in  this  study  (2  low-dose  males 
and  2  high-dose  females)  developed  hepatocellular  carcinoma. 
No  control  animals  exhibited  this  lesion.  Although  the 
incidence  of  this  malignant  liver  tumor  was  not  statistically 
significant,  these  results  may  have  biological  significance 
and  provide  reason  for  concern.  The  incidence  of 
hepatocellular  carcinoma  for  IBC-treated  male  rats  in  this 
study  was  1.9%,  while  the  incidence  for  treated  females  was 
2.1%.  In  published  reports  documenting  tumor  incidence, 
hepatocellular  carcinomas  occur  at  a  rate  of  0.2%  in  female 
and  0.8%  in  male  control  Fischer-344  rats  (11-16) .  However, 
the  increased  incidence  of  hepatocellular  carcinoma  did  not 
follow  a  dose-response  trend  characteristic  of  hepatocellular 
tumors  induced  by  chemical  carcinogens . 

Sixty-two  rats  in  this  study  developed  large  granular 
lymphocyte  leukemia,  a  common  neoplasm  of  lymphoid  tissue  in 
Fischer-344  rats.  This  disease  has  been  well-studied  and 
characterized  in  the  literature  (17,18,19).  The  incidence 
and  characteristics  of  this  disease  in  rats  used  in  this 
study  did  not  differ  significantly  from  published 
information.  Forty-two  rats  died  with  this  disease  prior  to 
scheduled  terminal  sacrifice.  The  premature  deaths  of  these 
animals  probably  had  little  or  no  impact  on  the  results  or 
conclusions  drawn  from  this  study. 

Most  of  the  rats  treated  with  IBC  had  gross  adhesions 
and  fibrous,  granulomatous  foreign-body  reactions  on  the 
liver  and  adjacent  organs  that  came  in  contact  with  the  test 
material.  These  lesions  persisted  for  the  normal  life-span 
of  the  rat.  As  noted  in  the  interim  report  covering  the 
first  year  of  this  study,  the  physical  presence  of  the 
compound  and  attendant  response  may  have  affected  the 
function  and  patency  of  the  intestinal  tract.  The  increased 
incidence  of  hepatocytic  vacuolization  in  livers  of  treated 
animals  indicates  possible  alterations  in  metabolism  of  these 
cells.  Although  such  altered  metabolism  could  be  due  to 
chemical  toxicity,  it  is  just  as  likely  the  result  of  local 
circulatory  disturbances  caused  by  the  presence  of  the 
compound  and  associated  foreign  body  reaction.  The  increased 
incidence  of  reactive  hyperplasia,  red  blood  cells,  and 
hemosiderin  pigment  in  lymph  nodes  of  treated  rats  is  not 
surprising.  Such  changes  normally  occur  in  nodes  draining 
areas  of  chronic  inflammation. 


Brown  et  al. — 34 


The  results  of  this  study  do  not  elucidate  fully  the 
carcinogenic  potential  of  isobutyl  2-cyanoacrylate.  Although 
the  data  suggest  the  sarcomas  were  induced  by  solid-state 
mechanisms,  they  are  not  definitive  because  of  a  lack  of  a 
dose  group  that  received  ground  polymerized  IBC.  The 
incidence  of  hepatocellular  carcinomas  in  treated  animals,  at 
a  rate  more  than  twice  that  reported  for  rats  of  the  same 
strain  and  age  in  other  studies  (although  a  statistically 
nonsignificant  incidence),  gives  cause  for  concern.  Because 
of  the  potential  usefulness  of  IBC  as  a  tissue  adhesive  in 
surgical  and  dental  procedures,  additional  studies  designed 
to  evaluate  fully  the  carcinogenic  potential  and  mechanisms 
of  this  material  should  be  conducted. 


CONCLUSION 

During  the  second  year  of  this  two-year  carcinogenicity 
bioassay  of  IBC,  there  were  no  treatment-related  changes  in 
survival,  weight  gain,  or  clinical  condition  of  the  rats. 
Compound-related  lesions  observed  in  animals  during  the 
second  year  included  adhesions  and/or  local  granulomatous 
reactions  of  the  liver  or  other  abdominal  organs,  and  liver 
or  other  soft  tissue  tumors  or  neoplasia.  An  increased 
incidence  of  sarcomas  was  attributed  to  the  chronic 
implantation  of  IBC  in  the  Fischer-344  rat.  There  was  no 
clear  evidence  that  the  hepatocellular  carcinomas  observed 
were  attributable  to  the  IBC. 


Brown  et  al. --35 


REFERENCES 

1.  Brown  LD,  Smith  CD,  Lollini  LO,  Korte  DW.  A 
carcinogenicity  bioassay  of  isobutyl  2-cyanoacrylate 
(IBC)  in  Fischer-344  rats — one-year  interim  sacrifice 
report.  Toxicology  Series  144.  Institute  Report  No. 

262.  Presidio  of  San  Francisco,  CA:  Letterman  Army 
Institute  of  Research,  March  1988. 

2.  National  Institutes  of  Health.  National  Research 
Council.  Guide  for  the  care  and  use  of  laboratory 
animals.  (5th  revised  ed.)  Bethesda,  MD :  U.S. 

Department  of  Health,  Education  and  Welfare,  1985;  NIH 
Publication  No.  85-23. 

3.  Chronic  bucrylate®  bioassay  procedures  within  toxicology 
GLP  suite  and  administrative  areas.  LAIR  Standard 
Operating  Procedure  OP-STX-81.  Letterman  Army  Institute 
of  Research,  Presidio  of  San  Francisco,  CA:  21  March 
1984  . 

4.  Chronic  carcinogen  bioassay.  LAIR  Standard  Operating 
Procedure  OP-STX-73.  Letterman  Army  Institute  of 
Research,  Presidio  of  San  Francisco,  CA:  9  November 
1983. 

5.  Food  and  Drug  Administration.  Nonclinical  laboratory 
studies,  good  laboratory  practice  regulations,  final 
rule.  In:  21  CFR  58.  Washington,  DC:  Food  and  Drug 
Administration,  22  Dec  1978;  43  FR  59986-60025. 

6.  Maronpot  RR,  Montgomery  CA,  Boorman  GA,  McConnell  EE. 
National  toxicology  program  nomenclature  for 
hepatoproliferative  lesions  of  rats.  Tox  Pathol  1986; 
14:263-273. 

7.  Turner  FC.  Sarcomas  at  sites  of  subcutaneously  implanted 
bakelite  discs  in  rats.  JNCI  1941;  2:81-83. 

8.  Bischoff  F,  Bryson  G.  Carcinogenesis  through  solid 
surfaces.  Progr  Exp  Tumor  Res  1964;  5:85-113. 

9.  Lawrence  WH.  Tumor  induction.  Chapter  17.  in:  von  Recum 
AF,  ed.  Handbook  of  biomaterials  evaluation:  scientific, 
technical,  and  clinical  testing  of  implant  materials. 

1st  ed.  New  York:  Macmillan  Publishing  Co,  1986:188-197 
(66  references) . 


Brown  et  al . — 36 


REFERENCES  (cont . ) 

10.  Woodward  SC,  Salthouse  TN.  The  tissue  response  to 

implants  and  its  evaluation  by  light  microscopy.  Chapter 
30.  In:  von  Recum  AF,  ed.  Handbook  of  biomaterials 
evaluation:  scientific,  technical,  and  clinical  testing 

of  implant  materials.  1st  ed.  New  York:  Macmillan 
Publishing  Co,  1986:364-378  (32  references). 

11.  Haseman  JK,  Huff  J,  Boorman  GA.  Use  of  historical 
control  data  in  carcinogenicity  studies  in  rodents.  Tox 
Pathol  1984;  12:126-135. 

12.  Haseman  JK,  Huff  J,  Rao  GN,  Boorman  GA,  McConnell  EE. 
Neoplasms  observed  in  untreated  and  corn  oil  control 
groups  of  F344/N  Rats  and  (C57BL/6N  x  C3H/HeN) FI  (B6C3F1) 
Mice.  JNCI  1985;  75:975-984. 

13.  Salleveld  HA,  Haseman  JK,  McConnell  EE.  Natural  history 
of  body  weight  gain,  survival,  and  neoplasia  in  the  F-344 
rat.  JNCI  1984;  72:929-940. 

14.  National  Institutes  of  Health.  National  Toxicology 
Program.  Toxicology  and  carcinogenesis  studies  of 
dimethyl  morpholinophosphoramidate  (CAS  No.  597-25-1)  in 
F344/N  rats  and  B6C3F1  mice  (gavage  studies) .  Research 
Triangle  Park,  NC:  US  Department  of  Health  and  Human 
Services,  January  1986;  Technical  Report  Series  NTP  No. 
298  (NIH  No.  86-2554)  . 

15.  National  Institutes  of  Health.  National  Toxicology 
Program.  Toxicology  and  carcinogenesis  studies  of  HC  red 
no.  3  (CAS  No.  2871-01-4)  in  F344/N  rats  and  B6C3F1  mice 
(gavage  studies) .  Research  Triangle  Park,  NC:  US 
Department  of  Health  and  Human  Services,  January  1986; 
Technical  Report  Series  NTP  No.  281  (NIH  No.  86-2537)  . 

16.  National  Institutes  of  Health.  National  Toxicology 
Program.  Toxicology  and  carcinogenesis  studies  of  Cl 
basic  red  9  monohydrochloride  (pararosaniline)  (CAS  No. 
569-61-9)  in  F344/N  rats  and  B6C3F1  mice  (feed  studies) . 
Research  Triangle  Park,  NC:  US  Department  of  Health  and 
Human  Services,  January  1986;  Technical  Report  Series  NTP 
No.  285  (NIH  No.  86-2541)  . 

17.  Ward  JM,  Reynolds  CW.  Large  granular  lymphocyte 
leukemia,  a  heterogeneous  lymphocytic  leukemia  in  F344 
rats.  Am  J  Pathol  1983;  111:1-10. 


Brown  et  al. --37 


REFERENCES  (cont . ) 

18.  Stromberg  PC.  Large  granular  lymphocyte  leukemia  in  F- 
344  rats.  Am  J  Pathol  1985;  119:517-519. 

19.  Losco  PE,  Ward  JM.  The  early  stage  of  large  granular 
lymphocyte  leukemia  in  the  F-344  rat.  Vet  Pathol  1984; 
21:286-291. 


Brown  et  al. — 38 


Page 

Appendix  A.  Chemical  Data . 39 

Appendix  B.  Animal  Data . 4  6 

Appendix  C.  Surgical  Report . 48 

Appendix  D.  Historical  Listing  of  Study  Events . 54 

Appendix  E.  Glossary  of  Terms  for 

Clinical  Observations . 57 

Appendix  F.  Individual  Animal  Histories . 64 

Appendix  G.  Gross  Necropsy  Observations, 

Incidence  Summary . 369 

Appendix  H.  Microscopic  Observations, 

Incidence  Summary . 37  9 

Appendix  I.  Glossary  of  Terms  for 

Pathology  Findings . 402 

Appendix  J.  Pathology  Individual  Animal  Data . 466 


Brown  et  al. --39 


Appendix  A:  CHEMICAL  DATA 

Chemical  Name:  Isobutyl  2-cyanoacrylate 

Other  Listed  Names:  Bucrylate®,  2-cyano-2-propenoic  acid  2- 
methyl-propyl  ester,  2-cyanoacrylic  acid  isobutyl  ester, 
bucrilate,  IBC,  IBCA* 

Chemical  Abstracts  Service  Registry  No.:  1069-55-2* 
Therapeutic  Category:  Surgical  aid  (tissue  adhesive)* 

LAIR  Code:  TP 60 
Chemical  Structure: 

COOH 

0=N 

Molecular  Formula:  C8H11N02 
Molecular  Weight:  153.18 
Physical  State:  Clear  colorless  liquid 
Boiling  Point:  170°C* 

Stability:  Stable  in  sealed  ampules.  Polymerizes  within 

minutes  on  contact  with  air.  Polymerizes  in  less 
than  1  second  on  contact  with  ionic  solutions, 
e.g.,  saline  or  blood.* 

Name  of  Contaminants  and  Percentages:  See  chemical  data  sheet 

attached 


CH, 

I  / 

HC— CH^-CH—C^ 


CH, 


Source:  Ethicon,  Inc. 

Somerville,  NJ  08876 

Lot  No.:  929-252 

Analytical  Data/Purity: 

Infrared  spectrophotometry  was  performed  on  11,  23,  and 
25  Jan  84,  and  the  results  were  identical  to  the 
standard  spectrum  from  Ethicon,  Inc.  Major  absorption 
peaks  were  observed  at  2965,  1740,  1470,  1385,  1290, 
1190,  980,  930,  and  715  cm"1. 


Brown  et  al. — 40 


Appendix  A  (cont.) :  CHEMICAL  DATA 

Analytical  Data/Stability: 

Samples  of  isobutyl  2-cyanoacrylate  (IBC)  were  evaluated 
by  gas  chromatography  on  11,  23,  and  25  Jan  84. t  Thus, 
IBC  was  analyzed  prior  to  dosing,  on  the  first  day  of 
dosing,  and  the  day  following  dosing.  The  chromatogram 
for  each  analysis  showed  only  one  peak,  with  a  retention 
time  of  3.3  min.  These  data  support  the  sponsor's  claim 
that  the  IBC  will  not  deteriorate  inside  the  sealed 
ampules . 


*  Windholz  M.  Merck  index.  10th  edition,  1983.  Monograph 
number  1433,  Bucrylate®.  Page  201-202.  Merck  &  Co,  Inc, 
Rahway,  NJ. 

t  O'Connor  RJ.  Memorandum  for  record,  26  Jan  1984,  Subject: 
Analysis  report,  GLP  Study  No.  83009,  Analytical  Chemistry 
Group.  Letterman  Army  Institute  of  Research,  Presidio  of 
San  Francisco,  CA. 


Brown  et  ai  . — 41 


Appendix  A  (cont . ) 


CHEMICAL  DATA 


ETHICON 

INC. 


SOMERVULE  .  NEW  JERSEY 

January  6,  1984 


To:  Dr.  W.  D.  Sheffield  cc:  Mrs.  S.  L.  Couchman 

Dr.  A.  W.  Fetter 
Mr.  J.  P.  O'Donnell 
Dr.  D.  W.  Regula 

Subject*  BUCRYLATE  TISSUE  ADHESIVE 
TUMORGENICITY  STUDY 


The  following  material  is  being  transmitted  to  you  for  initiation 
of  the  tumorgenicity  study: 

Lot  #929-252  (750  ampules) 

0.5  ml 
IBC-2 

One  hundred  twenty-five  ampules  have  been  included  for  stability 
assurance  testing  and  another  one  hundred  twenty-five  ampules  for 
long-term  retention  as  required  by  GLP  regulations. 

Attached  are  the  finished  goods  test  results  and  a  reference 
infrared  spectrum  for  this  lot.  The  primary  sterility  run  number 
is  R315001  and  the  overwrap  sterility  run  number  is  R343MB2. 

Please  let  me  know  if  I  can  answer  any  questions. 


mem 

Attachment 


Brown  et  al. — 42 


Appandix  A  (cont.):  CHEMICAL  DATA 


Finished  Goods  Test  Results 
Lot  #929-252 


Test 

Identification,  Infra-Red 
Total  Monomer,  %  (W/W) 

SO2,  ppm  (W/V) 

Hydroquinone,  %  -  (W/W) 

Isobutyl  Cyanoacetate,  ppm  (W/W) 
Color 

NIR  Absorptivity,  mg/absorbance  unit 
H2O,  ppm  (W/V) 

Heavy  Metals  as  Lead,  ppm  (W/W) 
Isobutanol ,  ppm  (W/V) 

Viscosity,  cps  at  30° C 


Resul t 

Conforms  to  standard 
99.9. 

562 

0.081 

None  detected 

0.046  at  400  nanometer 
0.0002  from  400-800  nanometers 
2.188 

<100 

<10 

725 

2.3 


\ 


Reference  #929-252 

Analytical  Service  Request  #20071 


Brown  et  al. — 43 


U 

13 

a 

o 

co 

CL 

a 

id 
X, 
c n 

►—4 

li. 

Id 

I 

CO 

Id 

I - * 

CO 


pQ 

in 

(\3 

i 


CD 

CVl 

CD 

I — 

a 

UJ 
i — 
<E 

>- 

u 

3 

CD 


Appendix  A  (confc.)  :  CHEMICAL  DATA  2 >/3 


a 

a 


HAVENUMBERS 


Brown  et  al. — 44 


Appendix  A  (cont.):  CHEMICAL  DATA 


reply  to 
attention  Of: 


SGRD-ULV-AC 


DEPARTMENT  OF  THE  ARMY 

letterman  army  institute  of  =-search 

PRESIDIO  OF  SAN  FRANCISCO,  CALIFORNIA  94t29 


26  January  1984 


MEMORANDUM  FOR  RECORD 

SUBJECT:  Analysis  Report,  GLP  Study  No.  83009,  Analytical  Chemistry  Group 


Date:  26  Jan  84 

Sample:  IBC-2  (Isobutyl“2-cyanoacrylate) 

Lot:  929-252 

I.  Identity 

The  Infrared  Spectrums  131,  132,  133  (11  Jan  84),  135,  136,  137  (23  Jan  84), 
and  138,  139,  140  (25  Jan  84)  are  consistent  with  the  Infrared  Spectrum  provided 
by  ETHICON  (SR  20071  -  12/22/83-DFB). 

Instrument:  Perkin-Elmer  Model  457 
Red  Tag  2912 
FSN  6650-C1 9-5014 

Methods:  Analytical  method  AD-100B,  identification  of  isobutyl-2- 
cyanoarcylates,  analytical  chemistry,  ETHICON,  Inc.  (9/16/70),  was 
used  as  the  basis  for  analysis*  The  Infrared  Spectrums  were  done 
in  two  different  ways*  The  first  followed  the  above  method  with 
the  following  exceptions:  KBr  (potassium  bromide)  was  used  for  the 
window  material  and  the  reference  cell  was  a  variable  path  length 
cell  that  was  matched  to  the  sample  cell.  The  second  method  was  a 
"Neat”  film  of  the  sample  between  KBr  windows*  The  film  metho£ 
provided  additional  information  including  a  peak  near  800  cm 
which  was  masked  by  the  solvent  in  the  first  method. 

II.  Purity 


The  gas  chromatographic  profiles  of  IBC-2  showed  one  major  component 
representation  1001  of  the  total  peak  area  when  it  was  run  on  11  Jan  84,  23  Jan 
84,  and  25  Jan  84. 

Methods:  Analytical  method  AD-100B,  identification  of  isobutyl-2- 
cyanoacralates ,  analytical  chemistry,  ETHICON,  Inc.  (9/16/70),  was 
used  as  the  basis  for  the  analysis. 


Brown  et  al.  —  45 


Appendix  A  (cont.) :  CHEMICAL  DATA 


SGRD-ULV— AC  26  January  1984 

SUBJECT:  Analysis  Report,  GLP  Study  No.  83009,  Analytical  Chemistry  Group 

Instrument:  Varian  4600 


Column:  3%  SP-2100  on  100/120 
Injector:  200°C 
Detector:  250°C 


Carrier  Gas:  Nitrogen  (20  cc/min) 
Run  Length:  15  minutes 
Detector:  FID 


Sensitivity:  10-11 
III.  Conclusion 

The  above  data  indicates  that  the  IBC-2,  Lot  929-252,  has  not  deteriorated 
in  the  sealed  vials  provided  by  ETHIC0N  and  is  consistent  with  the  data  provided. 


TvW\X' 


RICHARD  J.  O’CONNOR 
Research  Chemist 
Analytical  Chemistry  Group 


Brown  et  al. — 46 


Appendix  B :  ANIMAL  DATA 

Species:  Rattus  norvegicus 

Strain:  Fischer-344  (CDF) 

Source:  Charles  River  Breeding  Laboratories,  Inc. 

Wilmington,  MA  01887 

Reared  at  Kingston  (K62)  plant 

Sex:  Male  and  female 

Date  of  birth:  13  December  1983 

Method  of  randomization:  Weight  biased,  stratified  animal 

allocation  (TOXSYS  Animal 
Allocation  Program,  SOP  OP-ISG-24) 

Animals  in  each  group:  68  male  and  68  female  animals,  except 

female  control  group  totaled  67 
animals 

Condition  of  animals  at  start  of  study:  Normal 

Body  weight  range  at  dosing:  29  g  -  123  g 

(male  mean  «  94.0  g;  female  mean  =*  73.8  g) 

Toxicology  In-Life 

Identification  Procedures  (TOXSYS) : 

Ear-tagging  procedure  (SOP-OP-ARG  1) ,  tag  numbers 
84D00001  to  84D00209  (inclusive)  for  males  and 
84D00226  to  84D00435  (inclusive)  for  females. 

Pathology  Identification  Procedures: 

Animals  were  assigned  a  three-digit  number  for  data 
entry  into  the  Xybion  Data  Acquisition  Computer.  A 
cross  reference  list  for  the  TOXSYS  assigned  numbers 
was  generated  by  the  LAIR  Comparative  Pathology 
Branch  (Appendix  J) . 

Pretest  conditioning:  Quarantine/acclimation  11-22  January 

1984 

Justification:  The  Fischer-344  laboratory  rat  (versus  the 

Sprague-Dawley  or  Charles  River  CD  rat)  has  proven  to 
be  a  sensitive  and  reliable  model  for  chronic 
carcinogenesis  bioassays  due  to  its  low  incidence  of 
spontaneous  mammary  gland  and  liver  cancer.  This 
strain  is  recommended  by  the  NCI  Carcinogenesis 
Bioassay  Program. 


Brown  et  al. — 47 


Appendix  B  (cont . ) :  ANIMAL  DATA 


Group  Codes : 

Group 

Toxicology  Code 

Pathology  Code 

High-Dose  IBC 
(100  ^ll) 

A*  (Red) t 

1  § (High) 

Low-Dose  IBC 
(10  ^1) 

B*  (Yellow) t 

2  § (Low) 

Saline  Control 
(100  jxl) 

C*  (White) t 

3  § (CTL) 

*  Alphabetic  code  used  on  2  x  3  inch  cage  card  and  on 
animal's  chest  during  surgical  phase. 

t  Plastic  cage  card  protector  contained  a  bar-coded  animal 

number  (84D00 - )  and  a  0.5  inch  square  piece  of  color 

coded  tape. 

§  Numerical  code  used  to  group  pathology  data  within  Xybion 
computer  software. 


Brown  et  al. — 48 


SGRD-ULV-P 


Appendix  C:  SURGICAL  REPORT 


30  March  1984 


MEMORANDUM  FOR  RECORD 

SUBJECT:  Surgical  Report,  GLP  Study  83009 


1.  On  23  &  24  January  1984,  surgery  was  performed  in  the 
Operating  Room,  LAIR,  on  407  six-week-old  acclimated  Fischer- 
344  rats  assigned  to  the  Toxicology  Services  Group's  (TSG) 
chronic  two-year  Bucrylate®  Carcinogenesis  Bioassay. 

Surgeons  involved  were:  COL  Carpenter  and  LTC  Koppelman, 
USAIDR,  and  LTC  Rodkey,  LAIR.  Surgery  was  performed  under 
LAIR  GLP  Protocol  83009. 

2.  Surgery  was  necessary  to  enable  the  test  substance 
(Bucrylate)  to  be  applied  directly  to  the  surface  of  the 
liver  of  study  animals.  Surgery  on  179  males  occurred  on  23 
Jan,  and  surgery  on  the  remaining  25  males  and  203  females 
occurred  on  24  Jan  84 . 

3.  Animals  were  transported,  one  rack  of  60  rats  at  a  time, 
to  the  Operating  Room  Services  Group  (ORSG)  Surgical  Suite 
from  the  Toxicology  Services  Group  Animal  Suite.  Racks  were 
covered  with  sterile  sheets  to  provide  for  infection  control 
and  a  dark,  quiet,  unstressed  environment.  Surgery  started 
at  0730  hours  on  23  Jan  with  animal  number  84D00001  and 
progressed  through  animal  numbers  sequentially  to  84D00434 . 
Animals  were  randomly  assigned  to  the  three  dose  groups. 
Animals  were  presented  to  the  surgeons  in  a  random  fashion 
without  regard  to  group  assignment.  Animals  were  fasted 
(from  food  only)  1-4  hours  prior  to  surgery. 

4 .  Each  animal  was  examined  for  health,  checked  to  verify 
identification  (ear  tag,  cage  card  bar  code,  and  group 
assignment  tape),  and  weighed  on  an  Arbor  scale  (TOXSYS 
terminal  recorded) .  The  scale  was  recalibrated  each  morning 
prior  to  use.  Male  animals  weighed  approximately  100  g  and 
females  75  g  each.  Anesthesia  consisted  of  an  intramuscular 
hindlimb  injection  of  a  mixture  of  xylazine  HC1  (10  mg/kg) 
and  ketamine  HC1  (50  mg/kg) .  Anesthetic  was  prepared  by  LTC 
Rodkey  by  mixing  5  ml  ketamine  (100  mg/cc) ,  2 . 5  ml  Rompun® 
(20  mg/cc)  and  42.5  ml  of  sterile  saline.  Dose  calculations, 
lot,  and  expiration  of  anesthetics  and  diluent  are  provided 
in  Incl  #1. 


Brown  et  al. --49 


Appendix  C  (cont.):  SURGICAL  REPORT 

5.  Proper  identification  and  controls  on  group  assignment 
for  dosing  were  stressed.  Color  coded  tape,  containing  the 
animal's  number,  was  placed  on  the  tail  of  the  animal  while  a 
"back-up  system"  letter  signifying  dose  group  was  placed  on 
the  chest  of  the  animal  with  surgical  marker  pen  (black  CMS 
fine  tip  marking  pen,  lot  #138-800)  .  Animals  were  prepared 
for  surgery  by  clipping  the  abdomen  and  caudal  ventral  half 
of  the  thorax  with  a  number  40  Oster™  blade  and  Oster 
electric  clipper.  Animal  anesthesia,  weighing,  and  clipping 
were  performed  by  TSG  technicians  under  MAJ  Morgan's 
supervision. 

6.  Rats  were  placed  on  a  rat  restraining  board  in  the  supine 
position  and  the  clipped  area  was  scrubbed  with  povidone- 
iodine  surgical  soap  and  disinfected  with  povidone-iodine 
solution  and  alcohol.  The  rats  were  moved  to  the  surgery 
table  and  draped  so  that  only  the  surgically  prepared  area 
was  exposed.  LAIR  surgical  operating  procedures  applicable 
to  instrument  sterilization,  aseptic  technique,  and  sterile 
field  were  in  effect.  Surgical  packs  were  exchanged, 
cleaned,  and  autoclaved  after  every  6  animals.  Surgeons 
changed  gloves  after  every  six  animals,  and  dosing  personnel 
changed  gloves  frequently. 

7 .  A  midline  laparotomy  incision  was  made  extending  2-3  cm 
caudal  from  the  xyphoid  cartilage.  The  liver  was  exposed  by 
the  surgeon  using  either  ophthalmic  surgical  retractors  or  by 
"tenting"  the  abdominal  wall  with  forceps.  Bucrylate  (Lot 
929-252;  Ethicon,  Inc.)  or  sterile  saline  was  applied  to  the 
inferior  surface  of  the  exposed  liver  (usually  caudate  lobe) 
in  amounts  specified  in  the  protocol.  Dose  Group  1,  the  high 
dose  IBC  group,  received  100  microliters  (|il)  of  isobutyl  2- 
cyanoacrylate  (IBC);  Dose  Group  2  (low  dose  IBC)  received  10 
|il  of  IBC,  and  Group  3  (saline  control)  received  100  p.1  of 
sterile  isotonic  saline  (Lot  56-32 9-FD-05,  Exp.  1  Sep  1986; 
Abbott) .  Dosing  was  performed  by  TSG  personnel  using  fixed 
volume  Eppendorf  micropipettes  (calibrated  17  and  18  Jan  84) 
with  sterile  disposable  tips.  One  ampule  of  Bucrylate  was 
used  per  animal  for  dose  groups  1  and  2 — any  remaining  test 
compound  was  discarded. 

8.  One  animal  (84D00126)  was  misdosed  and  immediately 
removed  from  the  study.  After  application  of  the  IBC,  the 
abdominal  walls  were  retracted  or  "tented"  for  approximately 
two  minutes  to  allow  for  drying  of  the  test  substance. 

Dosing  was  monitored  by  the  principal  investigator,  MAJ  Brown 
(TOXSYS  ID  10186) . 


Brown  et  al . — 50 


Appendix  C  (cont.) :  SURGICAL  REPORT 

9.  The  laparotomy  incision  was  closed  using  two  layers.  The 
first  consisted  of  4-0  Vicryl®  (Ethicon,  Inc.)  in  a  simple 
continuous  pattern  for  the  abdominal  fascia,  muscle,  and 
peritoneum.  The  skin  was  closed  using  either  35  mm 
Proximate®  (Ethicon,  Inc.)  skin  staples  or  35  mm  Michelle 
skin  clips . 

10.  Animals  were  recovered  in  clean,  rat  rack  cages  in  the 
autoclave  room  adjoining  the  Surgical  Suite.  Room 
temperature  in  the  recovery  area  was  warm  (29 . 1-31 . 9°C)  . 

After  approximately  2  hours  in  the  recovery  area,  the  animals 
were  transferred  back  to  the  TSG  Animal  Suite,  and  given 
water  and  food. 

11.  Animals  were  monitored  postoperatively  by  LTC  Rodkey  and 
TSG  staff.  Skin  staples  were  removed  on  31  Jan  and  1  Feb  84 
(7-10  days)  by  TSG  veterinarians.  Twelve  of  the  407  rats 
died  or  were  removed  from  the  study  during  the  first  week 
after  surgery  due  to  underweight  condition,  surgical 
complications,  and/or  obstructed  intestine  due  to  Bucrylate 
adhesions.  Postoperatively,  rat  #84D00013  acquired  an 
infection  along  the  line  of  incision.  After  one  week  of 
treatment  with  hydrogen  peroxide,  the  incision  healed. 
Approximately  50  animals  were  noted  to  have  corneal  opacities 
one  week  postoperatively.  Keratoconjunctivitis  sicca  (dry 
eye)  and  subsequent  ulcerative  keratitis  are  fairly  common 
sequelae  to  rat  surgery.  The  etiology  is  thought  to  be 
related  to  drying  of  the  cornea  as  the  globe  protrudes,  and 
under  anesthesia,  the  eyelids  fail  to  cover  the  cornea  and 
the  blink  reflex  is  absent.  Ophthalmic  ointment  is  probably 
indicated  in  all  rodent  procedures  using  general  level 
anesthesia . 

12.  Overall,  the  surgery  went  very  smoothly  and  animals 
tolerated  the  procedure  well.  On  1  Feb  84,  382  animals  had 
recovered — the  initial  goal  was  to  have  at  least  360 
survivors  for  the  study. 

13.  Personnel  assisting  in  the  surgery  are  listed  in  Incl 
#2.  Study  records  are  on  file  listing,  by  animal,  the 
amounts  of  anesthesia  administered,  surgeon,  and  dosing 
personnel.  Ms.  Janice  M.  Adams,  Manager,  Regulatory  Affairs, 
Ethicon,  Inc.,  audited  the  surgery  on  23  &  24  Jan  84.  CPT 
Carroll,  LAIR,  QAO  also  visited  the  Surgical  Suite  during 
this  period. 


Brown  et  al.-~ 51 


Appendix  C  (cont.):  SURGICAL  REPORT 


14.  In  preparation  for  surgery,  a  meeting  was  held  between 
ORSG  and  TSG  personnel  on  13  Jan  84.  The  Ethicon  videotape 
on  the  surgical  procedure  was  reviewed  and  technicians 
practiced  the  dosing  procedure  using  Eppendorf  micropipettes 
and  one  ampule  of  Bucrylate.  Pilot  surgery  was  performed  on 
20  Jan  84 .  Eighteen  rats  were  operatively  laparotomized  and 
dosed  according  to  simulated  group  assignments. 


Incl 

as 


CF: 

COL  Carpenter 
LTC  Koppelman 
MAJ  Korte 

Raw  data  Binder,  83009 


Chief,  Operating  Room  Services  Group 
Diplomate,  Am  Coll  Vet  Surgeons 


I 


i 


Brown  et  al. — 52 


Appendix  C  (cont.):  SURGICAL  REPORT 

Enclosure  One  to  Surgical  Memorandum  For  Record,  30  March 
1984 


ANESTHESIA  GLP  STUDY  83009 


Rat  wt 

Xylazine* 

Ketaminet 

Vol  Mixtures 

80  g 

0.4  mg 

4  mg 

0.4  ml 

100  g 

0.5  mg 

5  mg 

0 . 5  ml 

120  g 

0 . 6  mg 

6  mg 

0 . 6  ml 

140  g 

0.7  mg 

7  mg 

0.7  ml 

*  Rompun®,  10  mg/cc,  Haver-Lockart ,  Shawnee,  KS. 
t  Vetalar®,  100  mg/cc,  Parke-Davis,  Morris  Plains,  NJ. 
§  Anesthetic  mixture  contained  10  mg  ketamine  and  1  mg 
xylazine  (Rompun®) /ml 


Mixtures  Components 

Xylazine  (Rompun®)  10  mg/kg,  20  mg/cc,  (Lot  260013,  Jan  85 
Exp. ) 

Ketamine  (Vetalar®)  50  mg/kg,  100  mg/cc,  (Lot  03793p,  Jul  88 
Exp . ) 

Saline  Diluent*,  (Lot  7C856X9,  Nov  84  Exp.) 


§  Anesthetic  Mixture  Formula:  5  ml  ketamine  soln  +  2.5  ml 
xylazine  soln  +  42.5  ml  saline  *  50  ml  anesthetic  mixture 

*  viaflex®,  0.9%  NaCl  Injection  USP,  Travenol 
Laboratories, Inc . ,  Deerfield,  IL. 


Brown  et  al. — 53 


Appendix  C  (cont.):  SURGICAL  REPORT 

Enclosure  Two  to  Surgical  Memorandum  For  Record,  30  March 
1984 


LAIR  PERSONNEL  WORKING  ON  SURGICAL  PHASE  OF  GLP  STUDY  83009 


Personnel 

Group 

Duty 

MAJ 

Korte,  Don  W.  Jr 

Toxicology 

Study  Director 

MAJ 

Brown,  Larry  D. 

Toxicology 

Principal 

Investigator 

MAJ 

Morgan,  Earl  W. 

Toxicology 

TOXSYS,  Presurgery 
Prep,  Recovery 

SFC 

Farmer,  Charles  N. 

Toxicology 

Scheduling 

SP5 

Kellner,  Thomas  P . 

Toxicology 

Dosing 

SP5 

Mullen,  Lawrence 

Toxicology 

Dosing 

SP5 

Rodriquez,  Justo 

Toxicology 

Prep 

PFC 

Sano,  Steven  K. 

Toxicology 

Prep,  Late  Night 
Recovery 

Ms  . 

Lewis,  Carolyn  M. 

Toxicology 

Dosing 

Ms . 

Coppes,  Valerie  G. 

Toxicology 

Initial  Recovery, 
Prep 

Mr. 

Dacey,  John 

Toxicology 

Dosing 

Mr . 

Spieler,  Richard  A. 

Toxicology 

Primary  Animal 
Caretaker 

Ms. 

Hernandez,  Susan 

Toxicology 

TOXSYS,  Prep 

Mr. 

Sands,  Edward  M. 

Toxicology 

Prep 

LTC 

Rodkey,  William  G. 

Operating  Rm 

Surgery 

SSG 

Del  la  Cerda,  Maria  V. 

Operating  Rm 

Surgery 

SSG 

Davis,  Garry 

Operating  Rm 

Surgery 

SP5 

Weber,  David 

Operating  Rm 

Surgery 

SP5 

Aiken,  Byron 

Operating  Rm 

Surgery 

SP5 

Cornier-Garcia  Juan 

Operating  Rm 

Surgery 

SP5 

Stevens,  Daniel 

Operating  Rm 

Surgery 

SP4 

Peterson,  Kimothy 

Operating  Rm 

Surgery 

PFC 

Rothhammer,  Gregory  A. 

Animal  Res 
Group 

Prep 

Brown  et  al. — 54 


Appendix  D 

Date 

11  Jan  84 

12  Jan  84 

11  -  22  Jan  84 

19  Jan  84 

20  Jan  84 

23  Jan  84  -  29 
Jan  85 

23  Jan  84 

24  Jan  84 

24  Jan  -  1 
Feb  84 

24,25,26,27, 
or  28  Jan  84 


HISTORICAL  LISTING  07  STUDY  EVENTS 

Event 

209  male  and  210  female  Charles  River 
(CDF)  Fischer-344  rats  were  received. 
Animals  were  checked  for  physical 
condition,  sexed,  individually  caged, 
and  fed. 

Rats  were  ear-tagged  and  weighed. 

Eight  rats  (4  male  and  4  female)  were 
submitted  for  necropsy  quality  control. 

Animals  were  observed  daily  during 
quarantine/acclimation  period. 

Animals  were  weighed  and  randomized 
into  dose  groups. 

Three  underweight  animals  and  one 
maloccluded  animal  were  sacrificed. 

All  animals  were  observed  twice  daily 
throughout  the  study  for  clinical  signs 
and  mortality. 

One  hundred  seventy-nine  animals  were 
weighed,  provided  with 
ketamine/xylazine  anesthesia, 
surgically  laparotomized,  and  implanted 
with  IBC  or  saline  according  to  dose 
group . 

Two  hundred  twenty-eight  animals  were 
weighed,  provided  with 
ketamine/xylazine  anesthesia, 
surgically  laparotomized,  and  implanted 
with  IBC  or  saline  according  to  dose 
group . 

All  animals  were  observed  frequently 
during  the  7-day  postoperative 
recovery  period. 

Twenty-five  animals  either  died  or  were 
sacrificed  during  the  postoperative 
period  because  of  complications  or 
underweight  conditions. 


Brown  et  al.  —  55 


Appendix  D  (cont . ) :  HISTORICAL  LISTING  OF  STUDY 

EVENTS 


Date 


Event 


1  Feb  84 


3  Feb  84  - 
27  Jan  85 


3,31  Mar,  8  Apr, 
9,18,19,27  Jun, 
14,16  Jul, 15  Sep, 
13  Dec  84 


12,19,23  or 

24  Jan  84, 

28  or  29  Jan  85 

16  or  17  Feb, 

15  or  16  Mar, 

12  or  13  Apr, 

10  or  11  May, 

7  or  8  Jun, 

5  or  6  Jul, 

2  or  3  Aug, 

30  or  31  Aug, 

27  or  28  Sep, 

25  or  26  Oct, 

22  or  23  Nov, 

20  or  21  Dec  84, 
and  17  or 
18  Jan  85 

28  -  29  Jan  85 


30  Jan  85 


30  Jan  85  -  10 
Feb  86 


Three  hundred  eighty-two  (93.9%)  of 
407  rats  that  went  to  surgery  remained 
on  study. 

Seventeen  unscheduled  animals  were 
necropsied  during  first  year  of  study  - 
14  sacrificed  due  to  poor  condition  and 
3  died  in  cage . 

Steam  outages  occurred,  or  animal 
suite  ventilation  fans  were  down. 

Spikes  in  room  temperature  and  small 
drops  in  relative  humidity  occurred 
during  these  periods. 

All  animals  were  weighed. 


All  animals  were  examined,  palpated  by 
veterinarian  or  toxicologist,  and 
weighed  at  28-day  intervals  (monthly) . 


Interim  sacrifice.  Sixty  animals  were 
weighed,  sacrificed,  and  necropsied. 

Three  hundred  five  rats  remain  "on 
study"  for  second  year  of  study. 

One  hundred  eighteen  unscheduled 
animals  were  necropsied  during  second 
year  of  study  -  97  sacrificed  due  to 
poor  condition  and  21  died  in  cage. 


Brown  et  al. — 56 


Appendix  D 


Date 

27  Apr,  21  Aug, 
26  Oct,  2  Nov, 

14  Dec  85,  7  Jan 


14  or  15  Feb, 

14  or  15  Mar, 

11  or  12  Apr, 

9  or  10  May, 

6  or  7  Jun, 

2  or  3  Jul, 

1  or  2  Aug, 

29  or  30  Aug, 

26  or  27  Sep, 

24  or  25  Oct, 

21  or  22  Nov, 

19  or  20  Dec  85, 
16  or  17  Jan  86 

27  Jan  -  14  Feb 


14  Feb  86 


(cont.):  HISTORICAL  LISTING  07  STUDY 

EVENTS 


ElYent 

Steam  outages  occurred,  or  animal  suite 
ventilation  fans  were  down.  Spikes  in 
86  relative  humidity  and  small  drops  in 

room  temperature  occurred  during  these 
periods . 

All  animals  were  examined,  palpated  by 
veterinarian  or  toxicologist,  and 
weighed  at  28-day  intervals  (monthly) . 


86  Final  sacrifice.  One  hundred  and 
eighty-seven  animals  were  weighed, 
sacrificed  and  necropsied. 

Study  "in-life”  completed. 


Brown  et  al.  —  57 


Appendix  E:  GLOSSARY  OF  TERMS  FOR  CLINICAL 

OBSERVATIONS 

Long-term  (TOXSYS®  Term)  -  Definition 

1.  General 

a.  Normal  (NORMAL)  -  animal  appears  to  be  healthy, 
exhibits  no  recognizable  clinical  signs. 

b.  Hunched  Posture  (HUNCH  POS)  -  an  increase  in  the 
curvature  of  the  thoracic  spine  as  viewed  from  the 
side . 

c.  Prostration  (PROSTRATE)  -  animal  stretched  out  in  a 
horizontal  position  but  conscious. 

d.  Loss  of  Consciousness  (LOSS  CONS)  -  prostrate  and 
unresponsive  to  sensory  stimuli  (touch,  sound,  pain, 
light,  etc. ) . 

e.  Moribund  (MORIBUND)  -  animal  is  near  death. 

f.  Death  (DEATH)  -  permanent  cessation  of  all  vital 
body  functions. 

g.  Emaciated  (EMACIATED)  -  the  animal  appears  to  have 
lost  weight,  but  the  skin  appears  hydrated  (i.e., 
the  skin  does  not  remain  in  place  when  squeezed 
together) . 

h.  Dehydrated  (DEHYDRATE)  -  the  animal  appears  to  have 
lost  weight  and  skin  remains  in  place  when  squeezed. 

i.  Pallor  (PALLOR)  -  a  lack  of  color  to  the  skin  or 
eyes . 

2 .  Behavioral 

a.  Inactive  (INACTIVE)  -  a  decrease  in  the  locomotor 
activity  of  the  animal. 

b.  Hyperactive  (HYPERACT)  -  an  increase  in  the 
locomotor  activity  of  the  animal. 

c.  Irritable  (IRRITABLE)  -  an  increase  in  annoyance  or 
anger  to  slight  stimulation. 


Brown  et  al. — 58 


Appendix  X  (cont . ) :  GLOSSARY  07  TERMS  FOR  CLINICAL 

OBSERVATIONS 


d.  Aggressive  (AGGRESS)  -  animal  is  bold,  wants  to 
fight,  characterized  by  a  desire  to  bite  for  no 
apparent  reason. 

e.  Vocalization  (VOCAL)  -  production  of  sound  from  the 
larynx . 

f.  Increased  or  Decreased  Preening  (IN  PREEN  or  DE 
PREEN)  -  an  increase  or  decrease  in  the  grooming 
behavior  of  the  animal. 

g.  Circling  (CIRCLING)  -  repeated  walking  or  running  in 
a  circle. 

h.  Jumping  (JUMPING)  -  springing  or  leaping  around  the 
cage . 

i.  Chewing  (CHEWING)  -  animal  goes  through  the  motion 
of  biting  and/or  grinding  with  no  external  stimulus. 

j .  Ataxia  (ATAXIA)  -  loss  of  coordination  characterized 
by  staggering  or  loss  of  gait. 

k.  Hypotonia  (HYPOTONIA)  -  decrease  in  muscle  tone. 

l.  Hypertonia  (HYPERTONIA)  -  increase  in  muscle  tone. 

m.  Catalepsy  (TRANCE)  -  animal  tends  to  remain  in  any 
position  in  which  it  is  placed. 

n .  Somnolence  (DROWSY)  -  animal  appears  drowsy,  but  can 
be  aroused  by  prodding  and  resumes  normal 
activities . 

o.  Tremors  (TREMORS)  -  a  trembling  or  quivering  of  the 
limbs  or  entire  body. 

p.  Fasciculation  (TWITCH)  -  twitching  of  the  muscles 
seen  on  the  back,  shoulders,  hind  limbs,  and  paws. 

q.  Writhing  (WRITH)  -  twisting  or  wrenching  the  body 
abnormally . 

r.  Paralysis  (PARALYSIS)  -  loss  or  impaired  motor 
function  in  the  limbs  or  entire  body. 


Brown  et  al. — 59 


Appendix  E  (cont.):  GLOSSARY  OF  TERMS  FOR  CLINICAL 

OBSERVATIONS 


s.  Disoriented  (DISORIENT)  -  animal  appears  dazed  and 
confused. 

3.  Convulsions 

a.  Tonic  Convulsion  (TONIC)  -  persistent  contraction  of 
muscles  with  rigid  extension  of  limbs. 

b.  Clonic  Convulsion  (CLONIC)  -  convulsion  alternating 
contraction  and  relaxation  of  muscles. 

c.  Tonic-clonic  Convulsion  (MIXED)  -  both  types  may 
appear  consecutively. 

d.  Opisthotonus  (OPISTOT)  -  tetanic  spasm  in  which  the 
back  is  arched  and  the  head  pulled  towards  the 
dorsal  position. 

4 .  Reflexes 

a.  Depressed  Myotactic  Reflex  (PULL)  -  decreased 
ability  of  animal  to  retract  its  hind  limb  when  the 
limb  is  pulled  down. 

b.  Depressed  Pinch  Reflex  (PINCH)  -  decreased  response 
(jerking  away  of  limb)  to  pinching  of  toes. 

c.  Depressed  Grasping  Reflex  (GRASP)  -  decreased 
ability  to  take  hold  of  finger  firmly  when  hung  from 
it. 

d.  Depressed  Righting  Reflex  (RIGHTING)  -  decreased 
ability  to  assume  normal  position  when  the  animal 
has  been  placed  on  its  side  or  back. 

e.  Depressed  Corneal  Reflex  (CORNEAL)  -  decreased 
response  (closing  of  the  eyelids)  to  touching  of  the 
cornea . 

f.  Depressed  Pupillary  Reflex  (PUPILLARY)  -  decreased 
constriction  of  the  pupil  in  the  presence  of  light. 

g.  Increased  Startle  Reflex  (STARTLE)  -  increased 
response  to  external  stimuli  (e.g.,  touch,  noise). 


Brown  et  al. — 60 


Appendix  X  (cont.):  GLOSSARY  OF  TERMS  FOR  CLINICAL 

OBSERVATIONS 


h.  Depressed  Pinnal  Reflex  (PINNAL)  -  decreased 

response  (twitching  of  external  ear)  elicited  by 
stroking  of  inside  surface  of  ear. 

5.  Respiratory 

a.  Increased  or  Decreased  Respiratory  Rate  (IN  RATE  or 
DE  RATE)  -  an  increase  or  decrease  in  the 
repetitions  of  inhaling  and  exhaling  air. 

b.  Increased  or  Decreased  Respiratory  Depth  (IN  DEPTH 
or  DE  DEPTH)  -  an  increase  or  decrease  in  the  volume 
of  air  inhaled  or  exhaled  with  each  repetition. 

c.  Irregular  Respiration  (IRREGULAR)  -  any  irregular 
rate  and/or  depth  of  respiration.  Since  this  can  be 
any  pattern,  a  description  should  be  given. 

d.  Apnea  (APNEA)  -  transient  cessation  of  breathing. 

e.  Gasping  (GASPING)  -  deep,  labored  inspiration,  often 
accompanied  by  a  wheezing  sound. 

f.  Panting  (PANTING)  -  rapid,  shallow  breathing. 

g.  Wheezing  (WHEEZE)  -  raspy  sound  made  with  breathing. 

h.  Sneezing  (SNEEZE)  -  animal  makes  sneezing-like 
sounds . 

i.  Nasal  Discharge  (NASAL  DIS)  -  a  discharge  from  the 
nose  is  produced.  A  description  of  the  color  is 
required. 

6.  Ocular 

a.  Lacrimation  (TEARING)  -  an  increase  in  the 
production  of  tears. 

b.  Chromodacryorrhea  (RED  TEARS)  -  production  of  red- 
colored  tears. 

c.  Miosis  (MIOSIS)  -  decreased  pupil  size. 

d.  Mydriasis  (MYDRIASIS)  -  increased  pupil  size. 


Brown  et  al. — 61 


Appendix  X  (cont.):  GLOSSARY  07  TERMS  FOR  CLINICAL 

OBSERVATIONS 


e.  Squinting  (SQUINTING)  -  closing  of  the  eyelids, 
usually  in  response  to  light. 

f.  Exophthalmus  (BULGING)  -  increased  protrusion  of  the 
eyeball . 

g.  Corneal  Opacity  (OPACITY)  -  an  opaque  spot  or  area 
on  the  cornea . 

h.  Conjunctivitis  (CONJUNC)  -  inflammation  of  the 
conjunctiva,  characterized  by  red,  swollen  eyelids. 

i.  Ptosis  (PTOSIS)  -  drooping  of  the  upper  eyelid  from 
paralysis . 

7.  Gastrointestinal 

a.  Increased  Salivation  (IN  SALIVA)  -  an  increase  in 
the  production  of  saliva. 

b.  Dried  Mucous  Membranes  (DRY  MOUTH)  -  drying  of  the 
mucous  membranes  in  the  mouth,  usually  from 
decreased  salivation. 

c.  Retching  (RETCHING)  -  a  strong,  unproductive  gag 
response . 

d.  Vomiting  (VOMITING)  -  the  expulsion  of  the  contents 
of  the  stomach  through  the  mouth. 

e.  Diarrhea  (DIARRHEA)  -  increased  frequency  and 
liquidation  of  fecal  discharges. 

f.  Scant  Feces  (SCANT)  -  decrease  in  the  frequency  of 
fecal  discharges. 

g.  Tarry  Feces  (TARRY)  -  blackish,  watery  feces  often 
due  to  blood  in  the  feces. 

8.  Skin/ fur 

a.  Hyperemia  (HYPEREMIA)  -  an  increase  in  reddish  color 
of  skin  over  entire  body,  usually  from  vasodilation 
(i .e . ,  flushed) . 


Brown  et  al. — 62 


Appendix  E  (cont . ) :  GLOSSARY  OF  TERMS  FOR  CLINICAL 

OBSERVATIONS 


b.  Cyanosis  (CYANOSIS)  -  a  bluish  discoloration  of  the 
skin,  often  related  to  respiratory  problems. 

c.  Jaundice  (JAUNDICE)  -  a  yellow  discoloration  of  the 
skin,  often  due  to  liver  dysfunction. 

d.  Scaling  (SCALING)  -  scaling  of  the  skin. 

e.  Erythema  (ERYTHEMA)  -  an  increase  in  the  redness  of 
the  skin  in  small  areas  (i.e.,  not  the  entire  body). 
Location  should  be  specified. 

f.  Bleeding  (BLEEDING)  -  loss  of  blood.  Location 
should  be  specified. 

g.  Scab  (SCAB)  -  a  crust  of  hardened  blood  and  serum 
over  a  wound  on  the  skin.  Location  should  be 
specified. 

h.  Edema  (EDEMA)  -  the  presence  of  large  amounts  of 
fluid  in  the  intercellular  tissue  spaces.  Location 
should  be  specified. 

i.  Ulceration  (ULCERATN)  -  the  formation  of  a  break  in 
the  skin  or  mucous  membrane  with  loss  of  surface 
tissue,  disintegration  and  necrosis  of  epithelial 
tissue,  and  often  pus.  Location  should  be 
specified. 

j.  Necrosis  (NECROSIS)  -  localized  death  of  living 
tissue.  Location  should  be  specified. 

k.  Alopecia  (HAIR  LOSS)  -  absence  or  loss  of  hair  from 
the  skin  where  it  normally  is  present .  Location 
should  be  specified. 

l.  Rough  Coat  (RGH  COAT)  -  a  rough  appearance  to  the 
coat  due  to  any  number  of  reasons,  such  as 
piloerection,  dullness,  lack  of  grooming. 

9.  Miscellaneous 

a.  Urine,  Feces  -  meant  to  be  used  with  color 

descriptors  to  describe  any  abnormal  coloration. 


Brown  et  al.  —  63 


Appendix  E  (cont . )  :  GLOSSARY  OF  TERMS  FOR  CLINICAL 

OBSERVATIONS 


b.  Stain,  Material  -  meant  to  be  used  with  color  and 
location  descriptors  to  describe  any  abnormal  stains 
or  material  found  on  the  coat . 

c.  Color  Descriptors  -  clear,  light,  dark,  yellow, 
orange,  red,  blue,  green,  and  brown. 

d.  Location  Descriptors  -  head,  mouth,  eye,  ear,  nose, 
neck,  front  leg,  hind  leg,  thorax,  abdomen,  back, 
side,  flank,  perianal,  tail,  left,  right,  and  both. 


Brown  et  al. — 64 


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STUDY:  0083009  CARCINOGENICITY  BIOASSAY  OF  ISOBUTYL  2-CYANO-  PAGE  22 

COMPOUND:  ISOBUTYL  2-CYANOACRYLATE  ACRYLATE  (IBC)  IN  MALE  AND  FEMALE  RATS.  DATE:  6/09/87 


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STUDY:  0083009  CARCINOGENICITY  BIOASSAY  OF  ISOBUTYL  2-CYANO-  PAGE  24 

COMPOUND:  ISOBUTYL  2-CYANOACRYLATE  ACRYLATE  (IBC)  IN  MALE  AND  FEMALE  RATS.  DATE:  6/09/87 


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(cont.):  INDIVIDUAL  ANIMAL  HISTORIES 


Brown  et  al . — 100 


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7/02/85  7:36:00  0000337  440.0  INTRAABDOMINAL  MASS  VENTRAL  TO  LIVER  SIZE  LARGE 
8/01/85  8:54:12  0001576  440.0  INTRAABDOMINAL  MASS  VENTRAL  TO  LIVER  SIZE  LARGE 
8/29/85  7:52:58  0000337  354.0  INTRAABDOMINAL  MASS  VENTRAL  TO  LIVER  SIZE  LARGE 
8/30/85  11:29:06  0000337  INTRAABDOMINAL  MASS  VENTRAL  TO  LIVER  SIZE  LARGE 


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(cont . ) :  INDIVIDUAL  ANIMAL  HISTORIES 


Brown  et  al. — 106 


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5/23/85  7:16:06  0000337  INTRAABDOMINAL  MASS  VENTRAL  TO  LIVER  SIZE  MEDIUM 

6/05/85  7:48:20  0000337  INTRAABDOMINAL  MASS  VENTRAL  TO  LIVER  SIZE  MEDIUM, HYPOTONIA  MODERATE, 

DECREASED  PREENING  MODERATE, ROUGH  COAT  MODERATE, STAIN  BROWN  NOSE  MODERATE 
6/05/85  7:57:52  0000337  *  No  observations  recorded. 


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Brown  et  al. — 215 


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Appendix  r  (cont.):  INDIVIDUAL  ANIMAL  BISTORIES 


Brown  et  al . 


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Brown  et  al. — 217 


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RIGHT  EYE  SLIGHT, EUTHANIZED 


Brown  et  al. — 222 

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COMPOUND:  ISOBUTYL  2-CYANOACRYLATE  ACRYLATE  (IBC)  IN  MALE  AND  FEMALE  RATS.  DATE:  6/09/87 


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COMPOUND:  ISOBUTYL  2 -CYANOACRYLATE  ACRYLATE  (IBC)  IN  MALE  AND  FEMALE  RATS.  DATE:  6/09/87 
START  DATE:  1/28/85  TIME:  14:40:13 

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(cont.):  INDIVIDUAL  ANIMAL  HISTORIES 


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COMPOUND:  ISOBUTYL  2-CYANOACRYLATE  ACRYLATE  (IBC)  IN  MALE  AND  FEMALE  RATS.  DATE:  6/09/87 
START  DATE.  1/28/85  TIME:  14:40:13 

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Brown  et  al. — 307 


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11/22/85  8:38:30  0000337  268.0  CORNEAL  OPACITY  BOTH  EYE  SLIGHT 
12/20/85  9:49:48  0000337  257.0  CORNEAL  OPACITY  BOTH  EYE  SLIGHT 
12/23/85  10:54:22  0000337  ■*  No  observations  recorded. 


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7/03/85  8:46:32  0000337  270.0  STAIN  BROWN  NOSE  SLIGHT 

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„TUDY:  0083009  CARCINOGENICITY  BIOASSAY  OF  ISOBOTYL  2-CYANO-  PAGE  141 

COMPOUND:  ISOBUTYL  2-CYANOACRYLATE  ACRYLATE  (IBC)  IN  MALE  AND  FEMALE  RATS.  DATE:  6/09/87 


Brown  et  al . — 366 


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6/07/85  10:26:30  0000337  246.0  NORMAL 

7/03/85  9:51:06  0000337  260.0  NORMAL 

7/09/85  12:47:12  0069406  *  No  observations  recorded 


Appendix  G  (cont.):  GROSS  NECROPSY  OBSERVATIONS,  INCIDENCE  SUMMARY 

IETTERHAN  ARHT  INSTITUTE  OF  RESEARCH  INCIDENCE  SUMMARY  REPORT  FOR  GROSS  NECROPSY  OBSERVATIONS  PRINTED:  21-NOV-89 

OIV  OF  RES  SUPP,  PATH  SERV  GP  STUDY  NUMBER:  GLP83009  PAGE:  3 

PRESIDIO  OF  SAN  FRANCISCO,  CA  94129  REPORT  FOR  UNSCHEDULED  DEATHS:  DAY  374  TO  DAY  755  OF  STUDY 


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TOTAL :  . 


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Appendix  6  (cont.):  GROSS  NECROPSY  OBSERVATIONS,  INCIDENCE  SUMMARY 

LETTERMAN  ARmV  INSTITUTE  Of  RESEARCH  INCIDENCE  SUMMARY  REPORT  fOR  GROSS  NECROPSY  OBSERVATIONS  PRINTED:  21-NOV-89 

01V  Of  RES  SUPP,  PATH  SERV  GP  STUDY  NUMBER:  GLP83009  PAGE:  4 

PRESIDIO  Of  SAN  fRANCISCO,  CA  94129  REPORT  fOR  ftNAL  SACRIFICE 


Brown  et  al . — 378 


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Appendix  H:  MICROSCOPIC  OBSERVATIONS,  INCIDENCE  SUMMARY 

LETTERMAN  ARMY  INSTITUTE  OF  RESEARCH  INCIDENCE  SUMMARY  OF  MICROSCOPIC  OBSERVAT IONSCALL  NEOPLASMS)  PRINTED:  22-NOV-89 

DIV  OIL  RES  SUPP,  PAIN  SERV  GP  STUDY  NUMBER:  GLP83009  PAGE:  1 

PRESIDIO  OF  SAN  FRANCISCO.  CA  94129  PATHOLOG I  ST ( S) :  MELLICK,  PAUL  U.,  Smith,  CBtherine  D 


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Appendix  H  (cont.):  MICROSCOPIC  OBSERVATIONS,  INCIDENCE  SUMMARY 

LETTERMAN  ARMY  INSTITUTE  OF  RESEARCH  INCIDENCE  SUMMARY  OF  MICROSCOPIC  OBSERVAT lONStMAL 1 CNANT  NEOPLASMS)  PRINTED:  22-NOV-69 

01V  OF  RES  SUPP,  PATH  SERV  CP  STUDY  NUMBER:  GLP83009  PACE:  2 

PRESIDIO  OF  SAN  FRANCISCO,  CA  94129  PATHOLOGI ST (S) :  MEL  LICK,  PAUL  U.,  Saith,  Catherine  D 

SPECIES:  RAT/FISCNER'344  STUDY  START  DATE:  23-JAN-84  STUOY  TYPE:  CHRONIC/2  YR  CARCINOGENIC 


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NOTE:  ENTRIES  FLAGGED  UITH  A  -  (MINUS)  ARE  SIGNIFICANTLY  DIFFERENT  FRON  CONTROL  AT  THE  0.05  LEVEL  USING  KOLMOGOROV- SMI RNOV 
TWO-TAILED  TEST. 


Appendix  H  (cont.):  MICROSCOPIC  OBSERVATIONS,  INCIDENCE  SUMMARY 

LETTERHAN  ARMY  INSTITUTE  Of  RESEARCH  INCIOENCE  SUMMARY  OF  MICROSCOPIC  OBSER VAT  I ONSIHAL I  CHANT  NEOPLASMS)  PRINTED:  22-NOV-B9 

DIV  OF  RES  SUPP,  PATH  SERV  CP  STUDY  NUMBER:  GLP83009  PAGE:  5 

PRESIOIO  OF  SAN  FRANCISCO,  CA  94129  PATHOLOG I  ST ( S) :  MELLICK,  PAUL  U.,  Snith,  Catherine  D 

SPECIES:  RAT/F1SCNER-344  STUDY  START  DATE:  23-JAN-84  STUDY  TYPE:  CHRONIC/2  YR  CARCINOGENIC 


Brown  et  al 


PRESENCE  OF  METASTATIC  SARCOMA 
PRESENCE  OF  METASTATIC  NEOPLASM 


Appendix  H  (cont.):  MICROSCOPIC  OBSERVATIONS ,  INCIDENCE  SUMMARY 

LETTERMAN  ARMY  INSTITUTE  Of  RESEARCH  INCIDENCE  SUNN ARY  OF  HICROSCOPIC  OBSERVAT IONS(BENIGN  NEOPLASMS)  PRINTED:  22-NOV-89 

OIV  OF  RES  SUPP,  PATH  SERV  GP  STUDY  NUMBER:  GLP83009  PAGE:  1 

PRESIDIO  Of  SAN  FRANCISCO,  CA  94129  PATHOLOGI ST (S) :  MELLICK,  PAUL  U.,  Saith,  Catherine  0 


brown  et  al 


NOTE:  ENTRIES  FLAGGED  WITH  A  •  (MINUS)  ARE  SIGNIFICANTLY  DIFFERENT  FROM  CONTROL  AT  THE  0.05  LEVEL  USING  KOLMOGOROV-SMI RNOV 
TWO-TAILED  TEST. 


Appendix  H  (cont.):  MICROSCOPIC  OBSERVATIONS,  INCIDENCE  SUMMARY 

IETTERMAN  ARMY  INSTITUTE  OF  RESEARCH  INCIDENCE  SUMMARY  OF  MICROSCOPIC  OBSERVAT IONS (BENIGN  NEOPLASMS)  PRINTED:  22-NOV-89 

01 V  OF  RES  SUPP.  PATH  SERV  GP  STUDY  NUMBER:  GLP83009  PAGE:  2 

PRESIOIO  OF  SAN  FRANCISCO,  CA  94129  PAT HOLOGI ST ( S) :  MELLICK,  PAUL  W.,  Smith,  Catherine  D 

SPECIES:  RAT/FISCNER-344  STUDY  START  DATE:  23-JAN-84  STUDY  TYPE:  CHRONIC/2  YR  CARCINOGENIC 


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TUO-TAILED  TEST. 


Appendix  H  (cont.):  MICROSCOPIC  OBSERVATIONS ,  INCIDENCE  SUMMARY 

LETTERMAN  ARMY  INSTITUTE  OF  RESEARCH  INCIOENCE  SUMMARY  OF  MICROSCOPIC  OBSERVAT IONS ( SEN  I  CM  NEOPLASMS)  PRINTED:  22-NOV-B9 

DIV  OF  R:  $  SUPP,  PATH  SERV  CP  STUDY  NUMBER:  GLP83009  PACE:  3 


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Appendix  B  (cont.):  MICROSCOPIC  OBSERVATIONS,  INCIDENCE  SUMMARY 

LETTERMAN  ARMY  INSTITUTE  Of  RESEARCH  INCIOENCE  SUMMARY  OF  MICROSCOPIC  OBSERVAT I ONS(BEN I CN  NEOPLASMS)  PRINTED:  22-NOV-89 

01 V  OF  RES  SUPP,  PATH  SERV  CP  STUDY  NUMBER:  GLPB3009  PACE:  4 

PRESIDIO  OF  SAN  FRANCISCO,  CA  94129  PAT HOLOG1 ST (S) :  HELL  I CK ,  PAUL  U.,  Snith,  Catherine  D 

SPECIES:  RAT/FISCHER*344  STUDY  START  DATE:  23-JAN-B4  STUDY  TYPE:  CHRONIC/2  YR  CARCINOGENIC 


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Appendix  H  (cont . ) :  MICROSCOPIC  OBSERVATIONS ,  INCIDENCE  SUMMARY 

LETTERMAN  ARMY  IHSTITUTE  Of  RESEARCH  IHCIOEHCE  SUMMARY  OF  MICROSCOPIC  OBSERVATIONS(NON-NEOPLASNS)  PRINTED:  22-N0V-89 

OIV  Of  RES  SUPP,  PATH  SERV  GP  STUDY  HUMBER:  GLP83009  PAGE:  2 

PRESIDIO  OF  SAH  FRANCISCO,  CA  94129  PAT HOLOG I  ST ( S ) :  MELLICK,  PAUL  U.,  Salth,  Catherine  D 

SPECIES:  RAT/FISCHER-344  STUDY  START  DATE:  23-JAN-84  STUDY  TYPE:  CHRONIC/2  YR  CARCINOGENIC 


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Appendix  H  (cont.):  MICROSCOPIC  OBSERVATIONS,  INCIDENCE  SUMMARY 

LETTERNAN  ARMY  INSTITUTE  OF  RESEARCH  INCIDENCE  SUMMARY  OF  MICROSCOPIC  OBSERVAT IONS(NON-NEOPL ASMS >  PRINTED:  22-NOV-B9 

DIV  OF  RES  SUPP,  PATH  SERV  OP  STUDY  NUMBER:  GLPB3009  PACE:  4 

PRESIDIO  OF  SAN  FRANCISCO,  CA  >4129  PAT NOIOGI ST < S ) :  NELLI CK ,  PAUL  U.,  Saith,  Catherine  D 


NOTE:  ENTRIES  FLAGGED  UITH  A  -  (MINUS)  ARE  SIGNIFICANTLY  DIFFERENT  FROM  CONTROL  AT  THE  0.05  LEVEL  USING  KOLMOGOROV - SM I RNOV 
TWO-TAILED  TEST. 


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TWO- TAILED  TEST 


Brown  et  al. — 402 


Appendix  I:  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 


GENERAL  DIAGNOSES 

In  this  study  there  were  a  few  lesions  which  occurred 
repeatedly  and  involved  numerous  organs  and  tissues.  The 
histologic  characteristics  of  these  lesions  had  very  little 
to  do  with  their  location  or  organ  of  involvement . 

Repetition  of  morphologic  descriptions  and  diagnostic 
criteria  for  these  lesions  under  each  organ  in  this  glossary 
would  be  redundant.  Therefore,  these  lesions  are  described 
here  under  the  heading  of  "General  Diagnoses,"  and  will  be 
referred  to  as  appropriate  under  the  organs  in  which  they 
occurred. 

1.  COMPOUND -RELATED  INFLAMMATORY  REACTION  -  The  test  material 
was  applied  directly  to  the  capsular  surface  of  the  liver  as 
a  liquid,  and  allowed  to  polymerize.  In  many  instances, 
especially  in  animals  in  the  high  dose  group,  the 
unpolymerized  material  ran  off  the  liver  and  adhered  to 
adjacent  abdominal  viscera  where  polymerization  occurred.  In 
histologic  sections,  the  test  material  is  a  clear,  slightly 
yellow  homogeneous  crystalline  material.  The  material 
stimulates  a  granulomatous  foreign-body  inflammatory  reaction 
characterized  by  infiltration  of  macrophages,  lymphocytes, 
and  large  multinucleated  foreign-body  giant  cells  within 
abundant  fibrous  connective  tissue.  Usually  this  lesion  is 
limited  to  the  serosal  surface  of  abdominal  viscera.  In  some 
cases,  there  is  necrosis  of  underlying  tissues  and 
penetration  of  the  serosal  surface  by  the  inflammatory 
response  accompanied  by  regenerative  hyperplasia  of  cells  of 
the  organ  involved.  The  most  probable  reason  for  the 
necrosis,  when  it  occurred,  is  the  local  compromise  of 
circulation  by  the  presence  of  the  test  material  or  the 
reaction  to  it.  No  attempt  is  made  to  grade  the  severity  of 
such  lesions.  They  are  simply  coded  as  "Present."  This 
diagnosis  is  used  only  when  there  is  unequivocal  evidence  of 
the  presence  of  test  material.  In  cases  where  test  material 
is  not  present,  a  different  diagnosis  such  as  "Fibrosis"  or 
"Chronic  Inflammation"  is  used,  as  appropriate. 

2.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  This  is  a  common 
neoplastic  disease  of  Fischer-344  rats.  Although  the  primary 
site  for  this  lesion  is  probably  the  spleen,  leukemic 
involvement  frequently  occurs  early  in  the  disease,  and 
neoplastic  lymphocytes  can  be  found  in  vascular  spaces  of 
virtually  every  organ  and  tissue  in  the  body.  The  neoplastic 
cells  are  large,  round  lymphoid  cells  with  abundant,  finely 


Brown  et  al. — 403 


Appendix  I  (cont.  )  :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

granular,  weakly  eosinophilic  cytoplasm.  Mitotic  figures, 
many  of  which  are  large  and  bizarre,  are  numerous.  Because 
of  the  characteristics  of  the  Xybion  computerized  data 
acquisition  system,  the  primary  site  for  this  neoplastic 
disease  is  coded  under  WHOLE  BODY,  thus  documenting  the 
presence  of  the  disease  regardless  of  the  organs  involved. 

It  is  coded  as  a  metastatic  lesion  only  in  those  organs  in 
which  it  alters  normal  histologic  architecture  or  constitutes 
a  significant  histologic  feature.  Using  tnis  method  of 
computer  entry,  it  is  possible  to  tabulate  the  occurrence  of 
this  disease  and  document  the  significant  organs  involved, 
without  causing  duplication  in  incidence  data. 

3.  PRESENCE  OF  METASTATIC  SARCOMA  -  A  number  of  the  animals 
developed  sarcomas  within  the  fibrous,  granulomatous  reaction 
incited  by  the  test  material .  The  sarcomas  involve  many 
organs  in  some  animals,  either  through  local  invasion  of 
malignant  cells  or  by  metastasis.  The  histologic 
characteristics  of  thes.e  tumors  are  extremely  variable.  The 
entire  spectrum  of  histologic  differentiation  of  mesenchymal 
tissue  is  represented  in  this  group  of  tumors.  Some  tumors 
are  well-differentiated  fibrosarcomas,  osteosarcomas,  or 
myxosarcomas,  while  others  are  very  anaplastic  and  poorly 
differentiated.  In  some  lesions,  multiple  types  of 
histologic  differentiation  occur  in  the  same  tumor.  The 
liver  is  the  most  frequent  location  for  these  lesions,  but  it 
is  not  involved  in  every  case.  Therefore,  the  primary  site 
for  these  lesions  is  coded  as  the  ABDOMINAL  CAVITY,  and  the 
involvement  of  each  individual  organ  is  coded  as  a  metastatic 
lesion.  Using  this  method  of  computer  entry  it  is  possible 
to  tabulate  the  occurrence  of  these  sarcomas  and  document  the 
organs  involved  without  causing  duplication  of  incidence 
data.  The  morphologic  characteristics  and  histologic 
differentiation  of  each  tumor  are  documented  under  the 
ABDOMINAL  CAVITY  using  the  "special  histologic  comments" 
feature  of  the  Xybion  program.  This  diagnosis  is  used  only 
when  it  is  certain  that  the  sarcoma  developed  within  the 
reaction  to  the  test  material .  Sarcomas  that  developed 
outside  the  abdominal  cavity  or  in  areas  remote  to  the 
application  of  the  test  material  are  not  included  under  this 
diagnosis . 

4.  PRESENCE  OF  METASTATIC  NEOPLASM  -  This  diagnosis  is  used 
in  cases  in  which  neoplastic  cells  (other  than  sarcoma  or 
large  granular  lymphocyte  leukemia)  are  present  in  organs  or 
tissues  other  than  those  of  the  site  of  origin.  The  lesion 


I 


Brown  et  al. — 404 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

is  coded  as  +  with  the  two-letter  abbreviation  of  the  organ 
of  origin.  Example:  an  adenocarcinoma  of  the  thyroid  that 
has  metastasized  to  the  lung  is  coded  under  lung  as  PRESENCE 
OF  METASTATIC  NEOPLASM  +TH. 

BRAIN 

1.  ASTROCYTOMA  -  A  neoplastic  proliferation  of  astrocytes. 

The  tumor  is  composed  of  large  and  medium-sized  cells  with 
large  vesicular  nuclei  and  cytoplasm  that  is  not  usually 
intensely  stained.  Cytoplasm  consists  of  numerous  fibrils 
which  are  tangled  and  crisscross  in  various  directions.  No 
particular  growth  pattern  is  characteristic.  These  tumors 
are  usually  small  and  well  demarcated  from  adjacent  tissue 
which  may  be  compressed  or  undergo  liquefaction  necrosis. 

2.  HYDROCEPHALUS  -  Enlargement  or  dilation  of  the  ventricles 
of  the  brain  at  the  expense  of  adjacent  parenchyma  which 
undergoes  pressure  atrophy  or  liquefaction  necrosis. 

3.  LIQUEFACTION  NECROSIS  -  A  necrotic  area  in  the  central 
nervous  system.  Necrotic  tissue  has  been  liquified.  The 
area  appears  microscopically  as  an  unlined  empty  space  with 
frayed  and  irregular  edges.  Cells  immediately  adjacent  to 
the  liquified  area  may  show  evidence  of  necrosis.  Pink- 
staining  proteinaceous  precipitate,  lipofuscin-laden  "gitter" 
cells,  red  blood  cells,  neutrophils,  and  remnants  of 
destroyed  tissue  may  or  may  not  be  present.  The  severity  of 
the  lesion  is  graded  based  on  size  of  the  necrotic  area: 

1  =  Diameter  is  less  than  than  that  of  a  40X 

microscopic  field. 

2  -  Diameter  is  less  than  a  20X  field. 

3  =  Diameter  is  less  than  a  10X  field. 

4  =  Diameter  is  less  than  a  4X  field. 

4.  PITUITARY  TUMOR  INVASION  -  Presence  of  neoplastic  cells 
from  the  anterior  pituitary  in,  or  adjacent  to  and 
compressing  brain  tissue.  These  tumors  do  not  usually 
metastasize  but  may  compress  and  destroy  brain  tissue 
locally.  Infiltration  of  meninges  and  ventricular  spaces  of 
the  brain  may  occur  and  is  included  under  this  diagnosis. 

5.  PERIVASCULAR  LYMPHOCYTIC  CELLULAR  INFILTRATION  -  Presence 
of  non-neoplastic  lymphocytes  in  perivascular  spaces. 


Brown  et  al. — 405 


Appendix  I  (coat.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

Severity  grade  criteria: 

1  -  Detectable. 

2  -  Involvement  of  an  area  up  to  size  of  20X 

microscopic  field. 

3  -  Cuffs  are  three  cell  layers  thick  and  involve 

area  greater  than  20X  microscopic  field. 

4  -  Cuffs  are  more  than  three  cells  thick  and/or 

area  of  involvement  is  greater  tnan  10X 
microscopic  field. 

6.  GLIOSIS  -  Non-neoplastic  proliferation  of  fibrous 
astrocytes  and  possibly  other  glial  cells.  Appears 
microscopically  as  a  focal  or  diffuse  increase  in  glial 
nuclei  and  increased  density  of  internuclear  areas.  This 
lesion  frequently  occurs  at  the  periphery  of  areas  of 
liquefaction  necrosis  but  is  not  limited  to  such  areas. 
Severity  grade  criteria: 

1  -  Detectable,  area  less  than  one  40X  field. 

2  =  Area  less  than  one  20X  field. 

3  -  Area  less  than  one  10X  field. 

4  -  Area  less  than  one  4X  field. 

5  -  Area  greater  than  one  4X  field. 

7.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
"General  Diagnoses"  above. 

8.  HISTIOCYTIC  INFILTRATION,  PERIVASCULAR  -  Presence  of  non- 
neoplastic  histiocytes  in  perivascular  spaces.  The  lesion  is 
coded  as  "Present . " 

XflACBSA 

Tracheal  glands  are  dilated  in  almost  every  animal 
examined  in  the  second  year  of  this  study.  Since  this  change 
is  universal  in  animals  in  all  treatment  groups,  it  is 
considered  a  normal  aging  change  for  this  population  and  is 
not  coded. 

1.  NONSUPPURATIVE  TRACHEITIS  -  Presence  of  mononuclear 
inflammatory  cells  within  or  beneath  the  tracheal  epithelium 
and/or  within  the  tracheal  wall. 


Brown  et  al . — 406 


Appendix  I  (cont.) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

Severity  grade  criteria: 

1  =  Presence  of  a  few  mononuclear  cells  within  or 

beneath  the  tracheal  epithelium. 

2  =  Inflammatory  cell  infiltration  of  the  tracheal 

mucosa  is  prominent.  Submucosal  glands  may  be 
infiltrated. 

3  =  Diffuse  infiltration  of  the  mucosa  and 

submucosa  with  inflammatory  cells  in  the 
tracheal  lumen. 

4  =  Same  as  Grade  3  plus  destruction  of  mucosal 

and/or  submucosal  structures . 

5  =  Inflammatory  infiltrate  and  destruction  of 

normal  structure  throughout  the  thickness  of 
the  tracheal  wall. 

2.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  "General  Diagnoses" 
above . 

3.  SUPPURATIVE  TRACHEITIS  -  An  acute  inflammatory  process 
characterized  primarily  by  neutrophils  within  the  trachea. 
Severity  grade  criteria: 

1  -  Presence  of  a  few  neutrophils  within  and 

beneath  the  tracheal  epithelium. 

2  *  Neutrophilic  infiltration  of  tracheal  mucosa 

and  submucosa  is  prominent .  Submucosal  glands 
may  be  infiltrated. 

3  =  Neutrophils  are  present  in  the  tracheal  lumen 

and  diffusely  infiltrate  mucosa  and  submucosa. 

4  *  Similar  to  Grade  3  plus  destruction  of  mucosal 

and  submucosal  structures. 

5  *  Neutrophilic  infiltrate  and  destruction  of 

normal  structure  throughout  the  thickness  of 
the  tracheal  wall. 

TSXRQID  fi LAMPS 

1.  FOLLICULAR  HYPERTROPHY/HYPERPLASIA  -  Non-neoplastic 
proliferation  of  thyroid  follicular  cells.  Cells  may  be 
increased  in  size,  number,  or  both.  Hyperplastic  epithelium 
may  form  follicles  containing  colloid,  or  form  papillary 
fronds  supporting  multiple  layers  of  cells  that  extend  into 
large  follicles.  The  lesion  is  coded  as  "Present." 

2 .  ADENOMA,  FOLLICULAR  -  A  benign  tumor  of  thyroid  follicular 
cells  characterized  primarily  by  cords  of  cells  or  cells 


Brown  et  al. — 407 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

forming  follicular  structures.  These  tumors  are  well- 
demarcated,  expansile  lesions  which  cause  compression  and 
atrophy  of  adjacent  thyroid  parenchyma. 

3.  PARAFOLLICULAR  CELL  HYPERPLASIA  (C-CELL  HYPERPLASIA)  - 
Focal  or  diffuse  non-neoplastic  proliferation  of 
parafollicular  cells.  Foci  of  hyperplastic  cells  are  no 
larger  than  a  normal  thyroid  follicle.  There  is  no 
compression  of  adjacent  thyroid  tissue..  Severity  grade 
criteria : 

1  =  One  to  three  nodules  or  less  than  25%  diffuse 

glandular  involvement . 

2  =  Three  to  six  nodules  or  less  than  50%  diffuse 

involvement. 

3  =  Six  to  ten  nodules  or  more  than  50%  diffuse 

involvement . 

4  =  More  than  ten  nodules  or  more  than  75%  diffuse 

involvement . 

4.  PARAFOLLICULAR  CELL  ADENOMA  -  Benign  tumor  of 
parafollicular  or  "C"  cells.  These  tumors  are  usually  solid 
in  cellular  arrangement,  and  are  large  enough  to  compress 
adjacent  tissue.  There  is  no  evidence  of  invasion  of  the 
thyroid  capsule  or  adjacent  tissues.  Cellular  pleomorphism 
may  be  prominent,  but  cells  are  usually  uniform  and  slightly 
elongated  with  ovoid  or  almost  spherical  nuclei.  Cytoplasm 
is  sparse,  homogeneous,  or  finely  granulated  and  stained 
weakly  eosinophilic.  Mitotic  figures  are  rare. 

5.  PARAFOLLICULAR  CELL  ADENOCARCINOMA  -  Malignant  neoplastic 
proliferation  of  parafollicular  or  "C"  cells.  Cellular 
morphology  may  vary  little  from  adenoma  described  above . 
Fusiform  and  palisade  patterns  occur  frequently.  Criteria 
for  malignancy  is  based  on  invasion  of  thyroid  capsule  and/or 
infiltration  of  adjacent  tissue  or  metastasis. 

6.  FOLLICULAR  ADENOCARCINOMA  -  Malignant  neoplastic 
proliferation  of  follicular  cells  of  the  thyroid. 

Microscopic  patterns  are  similar  to  those  of  follicular 
adenomas.  There  is  usually  very  little  structural  and 
cellular  pleomorphism,  and  moderate,  even  low  mitotic 
activity.  The  consistent  feature  that  distinguishes 
follicular  adenocarcinomas  from  the  analogous  type  of  benign 
tumor  is  their  invasiveness .  These  tumors  invade  the  thyroid 
capsule,  adjacent  tissues,  lymphatics,  and  blood  vessels. 


Brown  et  al . — 408 


Appendix  I  (cont . ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

7.  COLLOID  CYST  -  Presence  of  greatly  enlarged  (over  twice 
the  size  of  an  average  follicle)  colloid-filled  cystic  space 
within  the  thyroid  gland.  Epithelial  cells  lining  such  cysts 
are  usually  flattened.  One  or  more  colloid  cysts  may  occur 
within  a  single  thyroid  gland. 

PARATHYROID  GLANDS 

No  gross  or  microscopic  lesions  were  recognized  in 
parathyroid  glands  of  rats  used  in  this  study. 

ESOPHAGPS 

1.  NECROHEMORRHAGIC  ESOPHAGITIS  -  Necrosis  and  hemorrhage 
accompanied  by  acute  suppurative  inflammation  in  the 
esophagus.  This  was  an  uncommon  lesion  in  Study  GLP  83009. 
When  it  did  occur,  it  could  be  attributed  to  obstruction  of 
the  esophagus  by  test  material  and  attendant  granulomatous 
inflammatory  response  and  fibrosis  at,  or  near,  the 
esophageal  hiatus  of  the  diaphragm.  Severity  grade  criteria: 

1  =  Small  focal  area  of  necrosis  limited  to  the 

esophageal  mucosa  and  a  mild  local  neutrophilic 
infiltrate . 

2  -  Large  area  of  necrosis  extending  through  the 

mucosa  into  the  submucosa  with  prominent 
suppurative  exudate  and  hemorrhage. 

3  =  Similar  to  grade  2  but  necrosis  and  hemorrhage 

extend  into  the  muscular  wall  of  the  esophagus . 

4  =>  Necrosis,  hemorrhage,  and  suppurative  exudate 

extend  throughout  the  thickness  of  the 
esophageal  wall.  There  is  destruction  of  large 
amounts  of  esophageal  tissue  throughout  its 
thickness. 

SA1IVARX  fiLAHDS 

Parotid,  mandibular,  and  sublingual  glands  were  examined 
routinely.  The  gland  in  which  the  following  lesions  were 
observed  are  identified  by  notations  in  the  special 
histologic  comments  on  the  Xybion  program. 

1.  ATROPHY  AND  LOSS  OF  ACINAR  TISSUE  -  This  lesion  consists 
of  absence  of  acinar  secretory  cells.  Affected  lobules 
consist  of  ductal  structures  in  a  f ibrovascular  stroma.  Such 
lesions  are  usually  focal,  affecting  a  small  lobule  of 
salivary  tissue. 


Brown  et  al . — 409 


Appendix  I  (coat . ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

Severity  grade  criteria: 

1  =  One  lobule  affected. 

2  =  Two  or  three  lobules  affected. 

3  -  Four  to  six  lobules  affected. 

4  =  More  than  six  lobules  affected. 

2.  INFLAMMATION,  SUBACUTE  -  Presence  of  focal  or  diffuse 
infiltrations  of  inflammatory  cells  (primarily  lymphocytes, 
macrophages,  and  plasma  cells)  possibly  accompanied  by 
minimal  to  moderate  loss  of  glandular  tissue  through 
necrosis.  Severity  grade  criteria: 

1  -  One  to  three  foci  of  mononuclear  cells  no 

larger  than  the  diameter  of  a  secretory  acinus . 

2  =  Four  to  six  mononuclear  foci. 

3  -  Six  to  ten  inflammatory  cell  foci  or  up  to  two 

foci  of  necrosis  no  larger  than  the  diameter  of 
a  secretory  acinus . 

4  -  More  than  ten  inflammatory  cell  foci  and/or 

more  than  three  foci  of  necrosis  or  diffuse 
inflammatory  involvement  of  more  than  50%  of 
the  salivary  gland. 

3.  ADENOCARCINOMA  -  Malignant  neoplastic  growth  of  salivary 
glandular  or  ductal  epithelium.  Neoplastic  cells  are 
arranged  in  cords,  nests,  or  acini  often  forming  papillary 
fronds  or  large  glandular  spaces  filled  with  granular 
basophilic  secretory  product .  Cells  are  cuboidal  to  columnar 
with  pale-staining  granular  cytoplasm  and  round  to  slightly 
oval  vesicular  nuclei.  Neoplastic  cells  are  usually  rather 
well  differentiated  and  without  pleomorphism.  Mitotic 
figures  are  not  numerous.  A  small  amount  of  fibrous  stroma 
is  present  and  neoplastic  epithelial  cells  infiltrate  this 
tissue.  In  some  of  the  cystic  glandular  spaces  there  are  red 
blood  cells  and  bright.ly  eosinophilic  rhomboid  hemoglobin 
crystals . 

r.inrmt.  CT.awn _ fHARDERIAN  GLAND) 

1.  INFLAMMATION,  SUBACUTE  TO  CHRONIC  -  Presence  of  focal  or 
diffuse  infiltrations  of  lymphocytes,  macrophages,  and  plasma 
cells,  possibly  accompanied  by  minimal  to  moderate  loss  of 
glandular  tissue  through  either  necrosis  or  atrophy. 

Fibrosis  may  or  may  not  be  present.  In  most  cases  there  is 
intraductal  accumulation  of  inspissated  secretory  material. 


Brown  et  al. — 410 


Appendix  I  (cont . ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

Severity  grade  criteria: 

1  =  One  to  three  foci  of  mononuclear  cells  no 

larger  than  the  diameter  of  a  secretory  acinus. 

2  =  Four  to  six  mononuclear  foci;  fibrosis  may  be 

present . 

3  =  Six  to  ten  mononuclear  foci  with  or  without 

fibrosis  and  no  more  than  two  necrotic  foci  no 
larger  than  the  diameter  of  a  secretory  acinus . 

4  *  More  than  ten  inf lammatory/f ibrotic  foci  and/or 

three  or  more  foci  of  necrosis,  or  diffuse 
inflammation  and  fibrosis  of  more  than  50%  of 
the  gland. 

2.  ATROPHY  AND/OR  LOSS  OF  ACINAR  TISSUE  -  Absence  of  acinar 
secretory  cells.  Affected  lobules  consist  of  ductal 
structures  in  a  f ibrovascular  stroma.  Such  lesions  are 
usually  focal,  affecting  a  small  lobule  of  lacrimal  gland 
tissue.  Severity  grade  criteria: 

1  -  One  lobule  affected. 

2  *  Two  or  three  lobules  affected. 

3  =  Four  to  six  lobules  affected. 

4  -  More  than  six  lobules  affected. 

BXQRBITAIi LACRIMAL  filANP 

1 .  ACINAR  ATROPHY  -  Microscopic  appearance  and  grading 
criteria  are  identical  to  that  of  the  Harderian  gland 
described  above. 

2.  INFLAMMATION,  SUBACUTE  TO  CHRONIC  -  Microscopic  appearance 
and  grading  criteria  are  identical  to  that  of  the  Harderian 
gland  described  above. 

BBA&X 

1.  CARDIOMYOPATHY  -  This  is  a  very  common  lesion  in  aging 
Fischer-344  rats.  In  younger  animals,  it  is  usually  seen  as 
mild  to  moderate  focal  myocarditis.  As  the  rats  age,  the 
disease  progresses  to  myocardial  degeneration  characterized 
by  vacuolation  and  atrophy  of  myocardial  cells  followed  by 
fibrosis  and  loss  of  myocardial  fibers.  The  heart  is  not 
uniformly  affected  by  these  changes. 


Brown  et  al. — 411 


Appendix  Z  (coat . )  :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

Severity  grade  criteria: 

1  =  One  to  five  small  foci  of  lymphocytes  between 

myocardial  fibers  or  around  blood  vessels. 

2  =  Six  to  ten  foci  of  lymphocytes  and/or  minimal 

to  mild  vacuolar  change  in  myocardial  fibers. 
Fibrosis  may  be  present  but  is  limited  to  less 
than  25%  of  the  section. 

3  =  More  than  ten  foci  of  lymphocytes  and/or 

vacuolization  of  myofibers.  Mild  fibrosis 
involving  no  more  than  50%  of  the  section. 

4  *  More  than  half  of  the  section  is  involved  in 

degeneration  and  fibrosis.  Moderate  to  severe 
myofiber  atrophy  is  present. 

2.  ATR IOC AVAL  EPITHELIAL  MESOTHELIOMA  -  This  lesion  was  not 
observed  in  the  second  year  of  Study  GLP  83009.  In  the  first 
year  of  this  study,  one  rat  developed  this  unusual  tumor. 

The  lesion  was  completely  described  and  discussed  in  the 
interim  report  that  covered  the  first  year  of  this  two-year 
study . 

3.  MEDIAL  HYPERTROPHY  OF  MUSCULAR  ARTERIES  -  Thickening  of 
the  muscular  medial  layer  of  the  arteries  in  the  myocardium.  - 
Severity  grade  criteria: 

1  -  Minimal  detectable  thickening. 

2  *  Mild  thickening,  less  than  twice  normal 

thickness . 

3  ■  Moderate  thickening  of  the  media,  greater  than 

twice  the  normal  thickness,  often  producing 
distortion  of  arterial  lumen. 

4  ■  Marked  thickening  of  the  media,  approximately 

three  times  the  normal  thickness,  with  marked 
distortion  and  constriction  of  the  artsrial 
lumen . 

4.  MURAL  THROMBUS  -  Lesion  consists  of  a  mass  of  fibrin 
intermixed  with  red  blood  cells  and  neutrophils  attached  to 
the  endocardial  surface.  Adjacent  myocardial  cells  may 
undergo  degeneration  and  necrosis  and  infiltration  of 
inflammatory  cells.  The  lesion  is  graded  as  "Present." 

5.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  See  "General 
Diagnoses"  above. 


Brown  et  al. — 412 


Appendix  I  (cont.) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

6.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  "General  Diagnoses" 
above . 

lpngs 

1.  FOCAL  HYPERPLASTIC  PROLIFERATION  OF  TYPE  2  ALVEOLAR 
EPITHELIAL  CELLS  WITH  OR  WITHOUT  INFLAMMATORY  CELL 
INFILTRATION  -  Non-neoplast ic  proliferation  of  alveolar  type 
2  cells.  This  lesion  frequently  occurs  in  centriacinar 
regions  of  the  lung  at  the  junction  of  conducting  airways  and 
exchange  tissue.  It  also  occurs  near  pulmonary  adenomas  in 
alveoli  whose  ventilation  has  been  obstructed  by  the  tumor. 

In  these  areas,  alveolar  septae  are  lined  continuously  by 
type  2  epithelium.  Affected  alveoli  are  frequently 
collapsed.  Type  2  cell  proliferation  is  frequently 
accompanied  by  infiltration  of  alveolar  macrophages  both 
within  affected  alveolar  lumens  and  within  the  interstitium. 
Severity  grade  criteria: 

1  -  Focal  lesion  less  than  the  diameter  of  a  20X 

microscopic  field. 

2  -  Lesion  is  larger  than  the  diameter  of  a  20X 

field  but  smaller  than  that  of  a  4X  field. 

3  -  As  many  as  three  focal  lesions  with  diameters 

no  larger  than  that  of  a  4X  field  may  be 
present . 

4  -  More  than  four  focal  lesions  with  diameters  as 

large  as  a  4X  field  are  present. 

2.  PLEURITIS  WITH  SUBJACENT  CHRONIC  PNEUMONIA  -  Presence  of 
inflammatory  cells  and  fibrin  on  and  in  the  visceral  pleura. 
Inflammatory  cells,  fibrin,  and  possibly  the  replacement  of 
type  1  alveolar  epithelium  with  :ype  2  cells  are  present  in 
adjacent  pulmonary  parenchyma.  Severity  grade  criteria  for 
this  lesion  are  as  follows: 

1  -  Area  of  inflammation  is  less  than  the  diameter 

of  a  20X  microscopic  field. 

2  -  Area  of  inflammation  is  greater  than  a  20X 

field  but  less  than  a  10X  field. 

3  ■  Involvement  of  more  than  75%  of  a  lung  lobe. 

4  -  Involvement  of  the  entire  area  of  2  or  more 

lobes . 

3.  PULMONARY  EDEMA  -  Presence  of  protein-rich  (pink-staining) 
fluid  within  pulmonary  alveoli. 


Brown  et  al. — 413 


Appendix  I  (cont.):  GLOSSARY  OT  TERMS 

TOR  PATHOLOGY  FINDINGS 

Severity  grade  criteria: 

1  =  Edema  fluid  in  up  to  10  alveoli. 

2  =  Edema  fluid  in  10-50  alveoli. 

3  =  Edema  fluid  in  25%  to  50%  of  the  alveoli  in  one 

lobe . 

4  =  Edema  fluid  in  25%  to  50%  of  alveoli  in  2  or  3 

lobes . 

5  =  Edema  fluid  in  more  than  50%  of  the  alveoli  in 

all  lobes. 


4.  VASCULAR  CONGESTION  -  Pulmonary  vascular  spaces  are 
distended  with  large  amounts  of  blood.  Since  the  animals 
that  were  euthanized  were  killed  by  exsanguination  while 
under  barbiturate  anesthesia,  the  amount  of  blood  in  the 
pulmonary  vasculature  is  extremely  variable.  Animals  that 
died  naturally  have  much  more  blood  in  their  lungs.  For 
these  reasons,  vascular  congestion  was  coded  only  when  the 
congestion  constituted  a  prominent  histologic  characteristic 
of  the  lung. 

5.  THROMBUS  -  Presence  of  a  mass  of  fibrin  mixed  with  red 
blood  cells  and  leukocytes  attached  to  the  lumenal  wall  of  a 
vessel.  Tissues  supplied  by  the  thrombosed  vessel  may 
undergo  ischemic  necrosis  and  infiltration  of  inflammatory 
cells.  Severity  grade  criteria  for  this  lesion  are  as 
follows : 

1  =  Thrombus  is  present;  secondary  ischemic  changes 

involve  fewer  than  50  alveoli . 

2  -  Thrombus  is  present;  secondary  ischemic  changes 

involve  up  to  25%  of  a-  lung  lobe 

3  *  Thrombus  is  present  in  one  to  three  lobes  with 

ischemic  change  affecting  less  than  50%  of  the 
tissues  in  affected  lobes. 

4  -  Thrombus  is  present  in  three  or  more  lobes 

and/or  ischemic  change  affects  more  than  50%  of 
the  tissue  in  involved  lobes. 

6.  MINERAL I Z AT I ON  OF  PULMONARY  ARTERIES  -  Focal  dense 
deposits  of  mineral  within  the  wall  of  large  to  medium-sized 
arteries  in  the  lung.  Severity  grade  criteria  for  this 
lesion  are  as  follows: 

1  -  A  single  focus  of  mineral  which  produces  no 
distortion  of  the  vessel  wall.- 


Brown  et  al. — 414 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

2  *  Two  or  three  small  foci  of  mineralization  which 

produce  no  distortion  of  the  vessel  wall. 

3  =  Four  or  more  small  foci  of  mineral  which 

distort  the  vessel  wall  minimally,  or  one 
larger  focus  which  causes  moderate  distortion 
of  the  vessel  wall. 

4  =  More  than  one  large  focus  which  causes  moderate 

distortion  of  the  vessel  wall. 

7.  PNEUMONIA,  GRANULOMATOUS  -  The  infiltration  of 
lymphocytes,  macrophages,  and  neutrophils  with  variable 
numbers  of  large  multinucleated  giant  cells.  Necrosis  may  be 
present  in  the  center  of  granulomas.  These  lesions  are 
usually  focal  or  multifocal  and  are  located  within 
centriacinar  areas,  proximal  alveoli,  alveolar  ducts,  and 
terminal  bronchioles.  Severity  grade  criteria  for  this 
lesion  are  as  follows: 

1  =  Less  than  10%  of  the  lung  tissue  affected. 

2  =  10%  to  25%  of  the  lung  tissue  affected. 

3  =  25%  to  50%  of  the  lung  tissue  affected. 

4  =  More  than  50%  of  the  lung  tissue  is  affected. 

8.  PNEUMONIA,  INTERSTITIAL,  CHRONIC  -  Presence  of  chronic 
inflammation  within  the  pulmonary  parenchyma.  These 
inflammatory  lesions  usually  do  not  have  any  particular 
anatomic  orientation.  They  may  be  focal,  multifocal,  or 
diffuse.  Lesions  consist  of  infiltrations  of  macrophages, 
lymphocytes,  and  neutrophils  into  the  interstitium  and 
alveoli,  and  the  replacement  of  the  normal  mature  type  1 
alveolar  epithelial  cells  with  type  2  pneumocytes .  Fibrosis 
of  the  interstitium  may  be  present  in  older  lesions. 
Multinucleated  giant  cells  are  not  present.  Severity  grade 
criteria  for  this  lesion  are  as  follows: 

1  =  Less  than  10%  of  the  lung  tissue  affected. 

2  -  10%  to  25%  of  the  lung  tissue  affected. 

3  -  25%  to  50%  of  the  lung  tissue  affected. 

4  -  More  than  50%  of  the  lung  tissue  is  affected. 

9.  PERIVASCULAR  LYMPHOID  AGGREGATIONS  (CUFFING)  -  Presence  of 
lymphoid  tissue,  lymphocytes,  primitive  reticular  cells,  and 
plasma  cells  in  perivascular  connective  tissue.  This 
diagnosis  is  used  only  when  definite  vascular  orientation  of 
the  lymphoid  tissue  is  evident.  Since  severity  grading  of 
such  a  lesion  is  irrelevant,  the  lesion  is  coded  only  as 
being  "Present." 


Brown  et  al. — 415 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  7 I HD I MGS 

10.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  Presence 
of  neoplastic  lymphocytes  within  the  vascular  spaces, 
perivascular  tissues,  and  parenchyma  of  the  lung.  This 
lesion  is  coded  only  when  large  numbers  of  neoplastic  ceils 
constitute  a  prominent  histologic  feature  and/or  when  the 
normal  histologic  structure  of  the  lung  is  destroyed  or 
effaced  by  the  presence  of  neoplastic  cells. 

11.  PRESENCE  OF  METASTATIC  NEOPLASM  -  Presence  of  neoplastic 
cells  (other  than  lymphocytes  or  sarcoma  cells  which  have  a 
specific  diagnosis)  anywhere  within  the  lung.  Lesion  is 
coded  as  +  with  the  two-letter  abbreviation  for  the  organ  or 
tissue  of  the  primary  site.  Example:  +  TH  indicates 
metastatic  neoplasm  from  the  thyroid. 

12.  ALVEOLAR  HEMORRHAGE  -  Presence  of  red  blood  cells  within 
the  alveolar  spaces  of  the  lung.  In  almost  all  instances  in 
this  study,  this  lesion  is  caused  by  aspiration  of  blood  at 
the  time  of  euthanasia,  and  is  unrelated  to  the  experimental 
treatment .  Severity  grade  criteria  for  this  lesion  are  as 
follows : 


1  =  Blood  in  less  than  10  alveoli. 

2  =  Blood  in  10  to  50  alveoli. 

3  *  Blood  in  25%  of  the  alveoli. 

4  =  Blood  in  25%  to  50%  of  the  alveoli. 

13.  PERIBRONCHIAL  LYMPHOID  HYPERPLASIA  -  Normal  non¬ 
neoplastic  lymphoid  aggregates  were  present  in  peribronchial 
connective  tissues  of  nearly  every  animal  in  this  study. 

This  lesion  is  coded  only  when  the  amount  of  lymphoid  tissue 
appears  to  be  excessive.  Usually  in  such  cases  there  are 
other  inflammatory  lesions  in  the  lungs.  Severity  grade 
criteria  for  this  lesion  are  as  follows: 

1  *  Detectable  increase  in  lymphoid  tissue  limited 

to  airway  submucosa  and/or  peribronchial 
connective  tissue. 

2  -  Increase  in  submucosal  lymphoid  tissue  which 

extends  through  the  airway  epithelium. 

3  -  Increase  in  submucosal  and  peribronchial 

lymphoid  tissue  which  distorts  or  compresses 
airway  lumens  or  adjacent  parenchyma. 

4  =  Increase  in  lymphoid  tissue  which  causes 

moderate  to  marked  compression  of  airway  lumens 
and  adjacent  parenchyma. 


Brown  et  al . — 416 


Appendix  I  (cont .  )  :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

14 .  BRONCHIOLAR/ALVEOLAR  ADENOMA  -  Benign  tumor  of 
bronchiolar  epithelial  or  alveolar  epithelial  origin.  These 
lesions  are  usually  small  focal  proliferations  of  cuboidal  to 
columnar  epithelial  cells  which  compress  adjacent  pulmonary 
tissue.  Cells  are  frequently  arranged' in  long  serpentine 
cords.  Cells  that  are  usually  very  well  differentiated  and 
uniform  have  poorly  stained  clear  cytoplasm  and  small  basally 
located  ovoid  nuclei.  Few  mitotic  figures  are  present. 

15.  PRESENCE  OF  METASTATIC  SARCOMA  -  Presence  of  neoplastic 
mesenchymal  cells  anywhere  within  the  parenchyma  of  the  lung 
or  on  the  pleural  surface.  The  primary  site  for  these  tumors 
in  this  study  is  the  abdominal  cavity  where  the  tumors 
develop  in  the  fibrous  connective  tissue  constituting  part  of 
the  granulomatous  foreign-body  reaction  incited  by  the 
presence  of  the  test  material.  The  pulmonary  lesions  are 
coded  as  +  AC  indicating  presence  of  the  metastatic  sarcoma 
in  the  lung  with  the  primary  site  being  the  abdominal  cavity. 
Histologic  characteristics  of  the  tumor  are  documented  in 
special  histologic  comments  under  Abdominal  Cavity. 

xamia 

1.  PLEURITIS  -  Presence  of  acute,  subacute,  or  chronic 
inflammatory  reaction  on  the  surface  of  the  thymus.  This 
lesion  includes  the  infiltration  of  neutrophils,  mononuclear 
inflammatory  cells,  and  the  proliferation  of  fibroblasts  on 
the  surface  of  the  thymus .  Severity  grade  criteria  for  this 
lesion  are  as  follows: 

1  =  Inflammatory  lesion  less  than  the  diameter  of 

one  40X  microscopic  field. 

2  =  Inflammatory  lesion  less  than  the  diameter  of 

one  20X  microscopic  field. 

3  =  Inflammation  involves  25%  to  50%  of  the  thymus. 

4  =  Inflammation  involves  more  than  50%  of  the 

thymus . 

2.  ADENOMA,  ENDOCRINE  ORIGIN,  NOT  OTHERWISE  SPECIFIED  -  A 
small  (less  than  1  mm  in  diameter)  circumscribed 
proliferation  of  cells  characterized  by  finely  granular 
weakly  eosinophilic  cytoplasm  and  small,  round  to  ovoid 
nuclei.  Cells  are  arranged  in  small  nests  and  cords 
separated  by  delicate  f ibrovascular  stroma.  The  lesion  is 
situated  within  thymic  tissue.  Histologically,  the  cells 
comprising  this  small  adenoma  resemble  parafollicular 


Brown  et  al. — 417 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

("C"  cells)  of  the  thyroid  or  cells  from  the  parathyroid 
glands.  Carotid  or  aortic  body  origin  cannot  be  ruled  out. 
Only  one  such  lesion  was  observed  in  Study  GLP  83009. 

3.  THYMOMA  -  Benign  neoplastic  proliferation  of  thymic 
epithelial  cells.  The  lesion  is  a  well-demarcated,  un¬ 
encapsulated  proliferation  of  round  to  oval  cells  having 
eosinophilic  cytoplasm  with  poorly  demarcated  cellular 
boundaries  and  fairly  dense  round  to  ovoid  nuclei.  Mitotic 
figures  are  rare.  Cells  are  arranged  in  nests  and  bundles 
separated  by  very  fine  fibrous  stroma.  There  are  a  few 
widely  scattered  cells  or  clusters  of  cells  which  are 
partially  keratinized.  The  lesion  is  confined  to  one  fairly 
large  lobule  of  the  thymus.  There  was  only  lesion  of  this 
type  observed  in  Study  GLP  83009. 

4.  ADENOCARCINOMA,  NOT  OTHERWISE  SPECIFIED,  SITE  OF  ORIGIN 
UNDETERMINED  -  This  unusual  lesion  situated  within  the  thymus 
consists  primarily  of  proliferating  epithelial  cells  and  very 
few  lymphocytes.  Epithelial  cells  are  arranged  in  small 
nests  or  cords  with  some  papillary  forms.  The  lesion  is  well 
demarcated  from  adjacent  fat,  but  is  not  encapsulated.  Cells 
have  large  round  to  oval  vesicular  nuclei  and  abundant 
eosinophilic  cytoplasm  with  indistinct  plasma  membranes. 
Mitotic  figures  are  rare.  Scattered  throughout  the  tumor  are 
a  few  spaces  filled  with  eosinophilic  material,  a  few 
degenerating  epithelial  cells,  and  inflammatory  cells.  There 
was  only  one  lesion  of  this  type  observed  in  Study  GLP  83009. 

SPLEEN 

Because  of  the  method  of  euthanasia  used  in  this  study, 
the  amount  of  blood  in  the  spleen  varies  considerably.  Since 
this  feature  has  nothing  to  do  with  the  experimental 
treatment  or  toxic  effects  of  the  test  material,  no  diagnoses 
of  "splenic  congestion"  or  other  indication  of  the  amount  of 
blood  in  the  spleen  are  recorded. 

1.  LYMPHOID  HYPERPLASIA  OF  SPLENIC  CORPUSCLES  -  Non- 
neoplastic  proliferation  of  lymphoid  cells  in  splenic  white 
pulp  and  perifollicular  areas.  Severity  grade  criteria  for 
this  lesion  are  as  follows: 

1  =  Detectable  increase  in  diameter  of  splenic 
corpuscles  that  exceeds  that  of  the  average 
spleen. 


Brown  et  al. — 418 


Appendix  I  (cont.):  GLOSSARY  or  TERMS 

FOR  PATHOLOGY  FINDINGS 

2  ■  Increase  in  diameter  of  splenic  corpuscles  up 

to  twice  normal  size. 

3  ■  Increase  in  diameter  of  splenic  corpuscles  two 

to  three  times  normal  size. 

4  *  Marked  increase  in  diameter  of  splenic 

corpuscles  such  that  white  pulp  areas  are 
nearly  confluent. 

2.  FIBROSIS,  CAPSULAR  -  Non-neoplastic  proliferation  of 
fibrous  connective  tissue  in  the  splenic  capsule.  This  may 
be  part  of  an  adhesion  caused  by  the  test  material,  but  this 
diagnosis  is  not  used  when  test  material  is  present.  If  test 
material  is  present,  diagnosis  #11  below  is  used.  Severity 
grade  criteria  for  Fibrosis,  Capsular  are  as  follows: 

1  =  Focal  area  of  fibrosis  less  than  diameter  of 

one  40X  microscopic  field. 

2  =*>  Two  or  three  focal  areas  of  fibrosis,  similar 

to  those  in  grade  1,  or  one  area  involving  10% 
to  25%  of  the  surface  of  a  cross-section  of 
spleen . 

3  -  Fibrosis  involving  between  25%  to  50%  of  the 

surface  of  a  splenic  cross-section. 

4  -  Fibrosis  involving  more  than  50%  of  the  surface 

of  a  splenic  cross-section. 

3.  INFARCTION,  CHRONIC  -  Sharply  demarcated  area(s)  of 
splenic  parenchyma  comprised  primarily  of  fibrous  connective 
tissue  and/or  splenic  reticular  network  with  variable  numbers 
of  red  blood  cells  and  hemosiderin-laden  macrophages. 

Severity  grade  criteria  for  this  lesion  are  as  follows: 

1  =  One  focal  area  of  infarction  approximating  the 

area  of  one  40X  microscopic  field. 

2  »  One  or  more  areas  of  infarction  whose  total 

area  is  between  10%  to  25%  of  the  cross- 
sectional  area  of  the  spleen. 

3  -  One  or  more  areas  of  infarction  occupying  25% 

to  50%  of  the  cross-sectional  area  of  the 
spleen . 

4  -  One  or  more  areas  of  infarction  occupying  more 

than  50%  of  the  cross-sectional  area  of  the 
spleen . 

4.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  This 
common  neoplastic  disease  of  Fischer-344  rats  usually  begins 
in  the  spleen.  In  very  early  cases,  there  is  a  decreased 


Brown  et  al. — 413 


Appandix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

number  of  normal  lymphoid  cells  in  white  pulp  (lymphoid 
depletion) ,  and  proliferation  of  neoplastic  lymphocytes  in 
red  pulp  areas.  Neoplastic  cells  are  large,  round,  and  have 
abundant  finely  granular,  weakly  eosinophilic  cytoplasm  and 
large  round  vesicular  nuclei.  Mitotic  figures  are  numerous 
and  some  of  these  are  large  and  bizarre.  As  the  disease 
progresses,  the  numbers  of  neoplastic  cells  increase  greatly, 
causing  severe  splenic  enlargement.  Infarction  and  necrosis 
are  common  in  severely  affected  spleens.  Because  of  the 
features  of  the  Xybion®;  program,  the  primary  site  for  this 
entity  was  coded  under  Whole  Body,  and  each  organ  having 
significant  involvement  was  coded  as  having  a  metastatic 
lesion . 

5.  EXTRAMEDULLARY  HEMATOPOIESIS  -  In  most  of  the  animals  in 
this  study,  only  a  few  hematopoietic  cells  are  present  in  the 
spleen.  This  diagnosis  is  used  only  in  the  few  animals 
having  fairly  large  numbers  of  hematopoietic  cells  in  the 
spleen.  No  attempt  is  made  to  indicate  the  severity  of  this 
change.  The  lesion  is  indicated  as  "Present." 

6.  HEMOSIDERIN  PIGMENTATION  (RED  PULP)  -  Most  rats  in  this 
study  have  small  amounts  of  hemosiderin  pigment  in  splenic 
red  pulp.  However,  this  finding  is  coded  only  when  there 
appears  to  be  substantially  more  pigment  present  than  in  the 
average  rat  spleen.  The  change  is  graded  as  "Present"  with 
no  attempt  to  quantify  it  with  a  severity  grade. 

7.  SMOOTH  MUSCLE  HYPERTROPHY,  CAPSULE  AND  TRABECULAE  - 
Presence  of  increased  size  of  smooth  muscle  cells  within 
trabeculae  and  capsule  increasing  the  thickness  of  these 
structures.  Change  coded  as  "Present"  with  no  attempt  to 
grade  severity. 

8.  HISTIOCYTIC  INFILTRATE,  CAPSULE  AND  OMENTUM  -  Presence  of 
histiocytes  on  the  splenic  capsule  and  adjacent  omentum. 
Graded  as  "Present." 

9.  BENIGN  HYPERTROPHIC  NODULE  -  One  or  more  sharply 
demarcated  nodules  which  elevate  the  splenic  capsule  and 
consist  of  non-neoplastic  proliferation  of  lymphoid  cells. 
Some  of  these  structures  may  resemble  greatly  enlarged 
splenic  corpuscles  except  that  the  central  artery  is  usually 
absent.  This  lesion  is  graded  as  "Present." 


10.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  comments  and 


Brown  et  al. — 420 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

description  under  "General  Diagnoses"  above. 

11.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  comments  and 
description  above  under  "General  Diagnoses." 

12.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  comments  and 
description  above  under  "General  Diagnoses." 

LYMPH  NODES 

In  routine  histologic  evaluation  of  animals  from  this 
study,  the  mediastinal  nodes,  mesenteric  nodes,  and  nodes  in 
the  cervical  connective  tissue  adjacent  'to  salivary  glands 
(collectively  referred  to  as  "cervical  nodes")  were  examined. 
Diagnoses  for  all  nodes  are  coded  under  the  single  heading 
"lymph  nodes"  in  the  Xybion  computer  program.  The  specific 
nodes  having  a  coded  lesion  are  annotated  in  "special 
histologic  comments"  section  of  the  program. 

1.  LYMPHOID  HYPERPLASIA  -  Non-neoplast ic  proliferation  of 
lymphoid  cells  in  cortical  and  perifollicular  areas  of  the 
lymph  node.  Severity  grade  criteria  for  this  change  are  as 
follows : 

1  *  Detectable  increase  in  diameter  of  lymphoid 

follicles . 

2  =■  Increase  of  lymphoid  follicles  up  to  twice 

normal  size. 

3  =  Increase  of  lymphoid  follicles  to  two  or  three 

times  normal  size. 

4  -  Nearly  confluent  proliferation  of  lymphoid 

cells  in  cortex  of  the  node. 

2.  RED  BLOOD  CELLS,  HEMOSIDERIN  AND/OR  ERYTHROPHAGOCYTOSIS 
WITHIN  MEDULLARY  SINUSES  -  Finding  is  self-explanatory. 

Graded  as  "Present." 

3.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
description  and  comments  under  "General  Diagnoses"  above. 

4.  HISTIOCYTIC  INFILTRATION,  SUBCAPSULAR  AND  MEDULLARY 
SINUSES  -  Lymph  nodes  from  most  rats  in  this  study  have 
moderate  numbers  of  histiocytic  cells  in  medullary  areas. 

This  finding  is  coded  only  when  these  cells  are  very 
numerous.  The  change  is  coded  as  "Present." 


Brown  et  al. --421 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

5.  REACTIVE  HYPERPLASIA  —  Non-neoplastic,  noninflammatory 
increase  in  cellularity  in  lymph  nodes.  Lymphoid  follicles 
are  increased  in  size  and  number.  There  are  increased 
numbers  of  cells  in  perifollicular  areas  of  the  cortex. 
Medullary  cords  and  sinuses  frequently  contain  large  numbers 
of  plasma  cells.  Eosinophilic  protein-rich  edema  fluid  may 
be  present.  Change  is  coded  as  "Present." 

6.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  description  and 
comments  under  "General  Diagnosis"  above. 

7.  DILATED  AND/OR  CYSTIC  MEDULLARY  SINUSES  -  Self- 
explanatory.  Coded  as  "Present." 

8.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  description  and 
comments  under  "General  Diagnoses"  above. 

LIVER 

1.  COMPOUND  RELATED  INFLAMMATORY  REACTION  -  See  description 
and  comments  under  "General  Diagnoses"  above. 

2.  BILE  DUCTULE  EPITHELIAL  HYPERPLASIA/ CHOLANGIOFIBROSIS  - 
Non-neoplastic  proliferation  of  bile  ducts  frequently 
accompanied  by  periportal  fibrosis.  Fibrosis  is  considered  a 
part  of  this  lesion  and  is  not  coded  separately.  This  is  a 
very  frequently  observed  lesion  in  rats  in  this  study.  The 
change  is  usually  fairly  uniform  in  different  lobes  of  the 
liver.  Severity  grading  is  based  on  the  number  of  bile 
ductule  cross  sections  (lumens)  in  the  average  portal  triad: 

1  =  Three  lumens. 

2  *  Four  to  15  lumens . 

3  =  15  to  25  lumens. 

4  =  More  than  25  lumens. 

3.  HEPATITIS  -  Subacute  inflammatory  lesions  in  the  liver. 
These  lesions  are  usually  focal  or  multifocal  in  this  study. 
They  consist  of  small  focal  accumulations  of  macrophages, 
lymphocytes,  and  occasional  neutrophils  within  the  hepatic 
parenchyma.  Necrosis  or  degeneration  of  hepatocytes  within 
these  inflammatory  cell  foci  are  observed  occasionally. 
Severity  grade  criteria  are  based  on  the  number  of 
inflammatory  cell  foci  present: 

1  =  One  to  three  foci. 


Brown  et  al. — 422 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

2  -  Four  to  10  foci. 

3  *  Ten  to  20  foci. 

4  =  More  than  20  foci. 

4.  NON-NEOPLASTIC  FOCI  AND  AREAS  OF  CELLULAR  ALTERATION  - 
Three  distinctive  foci  or  areas  of  cellular  alteration  are 
present  in  livers  of  rats  in  this  study.  These  lesions  are 
named  for  the  morphologic  characteristics  of  their  component 
cell  populations,  e.g.,  basophilic  cell  foci,  clear  cell 
foci,  or  eosinophilic  cell  foci.  These  lesions  usually 
consist  of  a  uniform  cell  type,  although  they  may 
occasionally  contain  mixtures  of  cell  types.  Lesions  equal 
to  or  smaller  than  the  size  of  a  liver  lobule  are  foci  while 
a  lesion  larger  than  a  lobule  is  called  an  area.  Cellular 
arrangement  within  these  lesions  varies  little  from  normal 
hepatic  parenchyma.  The  cellular  appearance  and  staining 
characteristics  within  these  lesions  is  distinctly  different 
from  adjacent  normal  cells.  Hepatic  cords  of  the  lesion 
merge  imperceptibly  with  those  of  the  surrounding  tissue 
without  compression.  Foci  or  areas  of  cellular  alteration 
may  be  single  or  multiple,  and  different  morphologic  types 
may  occur  in  the  same  liver  section.  Their  distribution 
within  the  liver  is  not  uniform,  and  they  may  coexist  with 
any  other  type  of  hepatic  lesion. 

4A .  FOCUS  OF  CELLULAR  ALTERATION,  BASOPHILIC  CELLS  -  Cells 
are  usually  slightly  smaller  than  normal  and  cytoplasm  is 
distinctly  basophilic  in  hematoxylin  and  eosin  stained 
sections.  The  plates  of  hepatocytes  may  be  tortuous  due  to 
increased  numbers  of  cells. 

4B .  FOCUS  OF  CELLULAR  ALTERATION,  CLEAR  CELLS  -  A  sharply 
demarcated  focus  or  area  of  cells  having  a  clear  zone  around 
the  nucleus  in  which,  in  appropriately  stained  sections, 
glycogen  can  be  demonstrated.  Cells  may  be  of  normal  size  or 
enlarged. 

4C .  FOCUS  OF  CELLULAR  ALTERATION,  EOSINOPHILIC  CELLS  -  A 
well-demarcated  focus  or  area  of  hepatocytes  which  have 
acidophilic  cytoplasm  causing  a  "ground  glass"  appearance. 
Inclusions  may  be  present  in  the  cytoplasm.  Such  cells  are 
usually  enlarged  with  enlarged  nuclei  and  prominent  nucleoli. 
Cords  may  be  slightly  irregular.  Severity  grade  criteria  for 
these  lesions  are  as  follows: 


1  *  One  focus. 


Brown  et  al . — 423 


Appendix  Z  (cont . ) :  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

2  *  Two  to  four  foci,  or  one  area  alone  or  in 

combination  with  1  to  4  foci. 

3  =  Five  to  eight  foci,  or  two  to  three  areas. 

4  =  Eight  or  more  foci,  or  four  or  more  areas. 

5.  HEPATOCYTIC  VACUOLATION  -  Presence  of  small  to  large 
intracytoplasmic  vacuoles  within  hepatocytes .  In  sections 
appropriately  prepared  and  stained,  fat  can  be  demonstrated. 
Such  change  is  not  uniform  within  the  liver.  Severity  grade 
criteria  for  this  change  are  as  follows: 

1  =  Minimal  detectable  involvement. 

2  =  10%  to  20%  of  the  hepatocytes  are  vacuolated. 

3  *  20%  to  40%  of  the  hepatocytes  are  vacuolated. 

4  -  More  than  40%  of  the  hepatocytes  are 

vacuolated. 

6.  TELANGIECTASIS  -  Dilatation  of  hepatic  sinusoids.  Dilated 
sinusoids  are  usually  filled  with  blood.  Severity  grade 
criteria  for  this  lesion  are  as  follows: 

1  =  Focal  dilatation  of  sinusoids  occupying  a 

diameter  of  one  40X  microscopic  field. 

2  -  Widely  scattered  multiple  foci  occupying  an 

area  of  one  40X  microscopic  field. 

3  -  10%  to  20%  of  the  liver  section  affected. 

4  -  More  than  20%  of  the  liver  section  affected. 

7.  NECROSIS  -  Death  of  hepatocytes.  This  diagnosis  is  used 
to  indicate  necrosis  of  hepatocytes  without  any  particular 
anatomic  pattern.  Necrosis  is  designated  as  either  focal  or 
multifocal,  and  a  severity  grade  is  indicated.  See  Necrosis, 
Periacinar  and  Necrosis,  Hepatocytic,  Individual  Cells  below 
for  necrotic  lesions  having  specific  anatomic  patterns  of 
distribution.  Severity  grade  criteria  for  the  diagnosis. 
Necrosis,  are  as  follows: 

1  -  Necrotic  area  is  no  larger  than  a  40X 

microscopic  field. 

2  -  Combined  areas  of  focal  or  multifocal  necrotic 

lesions  are  less  than  10%  of  the  liver  section. 

3  -  Combined  areas  of  focal  or  multifocal  necrotic 

lesions  are  10%  to  20%  of  the  liver  section. 

4  =  Combined  areas  of  focal  or  multifocal  necrotic 

lesions  are  20%  to  40%  of  the  liver  section. 


Brown  et  al. — 424 


Appendix  I  (cont.):  GLOSSARY  or  TERMS 

FOR  PATHOLOGY  FINDINGS 

5  -  Combined  areas  of  focal  or  multifocal  necrotic 

lesions  are  greater  than  40%  of  the  liver 

section . 

8.  FIBROSIS,  CAPSULAR,  (NO  COMPOUND)  -  Presence  of  increased 
fibrous  connective  tissue  on  the  capsule  of  the  liver.  There 
is  minimal  inflammatory  cell  infiltration  and  no  foreign-body 
giant  cells.  This  lesion  is  coded  as  "Present." 

9.  PERIPORTAL  MONONUCLEAR  CELLULAR  INFILTRATION  -  Presence  of 
lymphocytes,  macrophages,  and  plasma  cells  in  the  periportal 
connective  tissue.  Severity  grade  criteria  for  this  lesion 
are  as  follows: 

1  =  Small,  widely  scattered  foci  not  to  exceed  more 

than  three  foci  per  liver  section. 

2  =  Four  to  six  small  foci  of  mononuclear  cells. 

3  *  Six  to  10  small  foci  or  presence  of  three  or 

more  foci  that  are  as  large  as  a  liver  lobule. 

4  =  More  than  ten  small  foci  or  more  than  four 

large  foci . 

10.  CYSTIC  DEGENERATION  -  Presence  of  focal  or  multifocal 
areas  characterized  by  cysts  with  pale  eosinophilic  amorphous 
material,  hepatocytes  that  are  enlarged  by  the  presence  of 
clear  cytoplasmic  vacuoles,  and  an  occasional  necrotic 
hepatocyte.  There  may  be  slight  peripheral  compression  of 
adjacent  hepatic  parenchyma.  The  lesion  is  coded  as 
"Present . " 

11.  HEPATOCYTIC  DEGENERATIVE  CHANGES  (NON-NECROTIC)  - 
Presence  of  diffuse  areas  in  which  hepatocytes  show  some  or 
all  of  the  following  changes:  cellular  enlargement,  nuclear 
enlargement,  nuclear  hyperchromasia,  multiple  nuclei, 
microvesicular  cytoplasmic  change.  Affected  hepatocytes  are 
in  normal  arrangement  and  necrosis  is  not  present.  This 
change  occurs  most  frequently  in  livers  severely  affected 
with  large  granular  lymphocyte  leukemia.  Severity  grade 
criteria  are  based  on  both  the  number  of  hepatocytes  affected 
(percent  of  sectional  area)  and  the  severity  of  the 
cytological  abnormalities  listed  above: 

1  -  Minimal  cytological  abnormality  affecting  less 

than  10%  of  the  section. 

2  -  Mild  cytological  abnormalities  and/or  more  than 

20%  of  the  section  affected. 


Brown  et  al . — 425 


Appendix  I  (cont. ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

3  =  Moderate  cytological  abnormalities  and/or  more 

than  40%  of  the  section  affected. 

4  =  Marked  cytological  abnormalities  and/or  more 

than  50%  of  the  section  affected. 

12.  NECROSIS,  HEPATOCYTIC,  INDIVIDUAL  CELLS  -  Presence  of 
coagulative  necrosis  of  individual  hepatocytes .  There  is  no 
specific  anatomic  orientation  or  pattern  to  the  necrotic 
lesion.  This  lesion  is  most  frequently  observed  in  cases  of 
large  granular  lymphocyte  leukemia  or  the  nonspecific 
multifocal  hepatitis  described  in  diagnosis  #3  above. 
Occasionally,  necrotic  cells  are  present  within  areas  of 
hepatocytic  vacuolization  and  adjacent  to  areas  of  compound- 
related  inflammatory  reaction.  Severity  grade  criteria  are 
based  on  the  number  of  necrotic  hepatocytes  per  high  power 
microscopic  field: 

1  =  One  to  three  necrotic  cells  per  high  power 

field.  The  area  affected  is  limited  to  that  of 
a  20X  field. 

2  =  One  to  three  necrotic  cells  per  high  power 

field,  with  large  diffuse  areas  of  involvement. 

3  *  Four  to  six  necrotic  hepatocytes  per  high  power 

field;  diffuse  area  of  involvement. 

4  -  More  than  seven  necrotic  hepatocytes  per  high 

power  field;  diffuse  involvement.  If  necrosis 
involves  foci  of  hepatocytes  rather  than 
individual  cells,  the  diagnosis  of  NECROSIS  (#7 
above)  is  used  with  appropriate  distribution 
modifier  and  severity  grade. 

13.  BENIGN  HYPERPLASTIC  NODULE (S)  -  Spherical  proliferation 
of  hepatocytes  without  cytologic  atypia.  Normal  hepatic 
architecture  is  present  but  it  may  be  distorted;  portal 
triads  may  be  present .  There  is  mild  compression  of  adjacent 
parenchyma.  Hepatocytes  within  a  focus  of  hyperplasia  are 
usually  uniform  and  have  a  homogeneous  growth  pattern,  but 
features  such  as  hypertrophy,  cytoplasmic  vacuolation,  and 
increased  numbers  of  mitoses  may  be  present.  Often  the 
distinction  between  focal  hyperplasia  and  a  focus  of  cellular 
alteration  rests  solely  on  the  degree  of  cytologic  alteration 
and  compression  of  adjacent  parenchyma.  Hyperplastic  lesions 
may  reach  several  millimeters  in  diameter  and  are  frequently 
seen  as  relatively  distinct  lesions  in'  histologic  sections. 
The  lesion  is  coded  as  "Present"  with  an  indication  of 
multifocal  distribution  if  multiple  lesions  are  present. 


Brown  et  al. — 426 


Appendix  I  (cont. ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

14 .  NECROSIS/  PERIACINAR  -  Coagulative  necrosis  of 
hepatocytes  with  a  definite  orientation  around  central  veins. 
In  more  severe  cases  the  necrotic  change  may  bridge  from 
central  vein  to  central  vein,  or  from  central  vein  to  portal 
areas.  Stromal  collapse  may  occur;  the  necrotic  area  may  be 
filled  with  blood.  Variable  numbers  and  types  of 
inflammatory  cells  may  be  present.  Adjacent  lobules  and/or 
the  capsule  may  be  distorted.  Changes -are  usually  uniform 
with  most  or  all  the  lobules  affected.  Severity  grade 
criteria: 

1  =  Minimal  detectable  necrosis  around  central 

veins . 

2  =  Definite  necrosis  involving  nearly  all 

periacinar  hepatocytes  in  nearly  all  lobules. 

3  =  Bridging  of  periacinar  areas  by  necrotic  cells 

and  accompanying  inflammatory  cells . 

4  =  Necrosis  and  bridging  of  periacinar  areas  with 

stromal  collapse  and  blood  pooling. 

15.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
comments  and  description  under  "General  Diagnoses"  above. 

16.  FIBROSIS,  PARENCHYMAL  (SCARRING)  -  Presence  of  fibrous 
connective  tissue  and  minimal  inflammatory  cell  infiltrate 
within  the  liver  parenchyma.  This  lesion  occurs  most 
frequently  near  areas  of  compound-related  inflammatory 
reaction.  Severity  grade  criteria: 

1  =  An  area  of  fibrosis  no  larger  than  that  of  a 

40X  microscopic  field. 

2  =  A  fibrotic  area  as  large  as  a  20X  microscopic 

field. 

3  =  A  fibrotic  area  as  large  as  2  to  4  liver 

lobules . 

4  *  A  fibrotic  area  larger  than  5  liver  lobules. 

17.  HEPATOCYTE  COALESCING  MICROVESICULAR  (FATTY)  CHANGE 
LEADING  TO  NECROSIS  AND  MINERALIZATION  -  Microscopically, 
this  lesion  consists  of  microvesicular  cytoplasmic  change  in 
hepatocytes,  rupture  and  disintegration  of  affected  cells, 
consequent  enlargement  of  adjacent  sinusoids,  and  the 
presence  of  nearly  colorless  to  light  blue  mineralized 
crystals.  The  lesion  usually  has  a  focal  or  multifocal 
distribution.  Frozen  sections  stained  with  oil-red-0  or 
Sudan  Black  demonstrate  the  presence  of  fat  in  vacuolated 
hepatocytes  near  the  dilated  sinusoids.  Numerous  crystals 


Brown  et  al . — 427 


Appendix  I  (cont.) :  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

are  present  in  these  areas.  Sections  stained  with  Alizarin 
Red-S  and  Von  Kossa  stain  demonstrate  that  the  crystals 
contain  both  calcium  and  phosphate.  Periodic  Acid-Schiff 
stains  are  noncontributory.  The  interpretation  of  these 
findings  is  that  fatty  degenerative  changes  in  hepatocytes 
lead  to  necrosis  and  subsequent  saponification  of  the 
accumulated  lipid.  Severity  grade  criteria: 

1  =  Changes  occur  in  small  widely  scattered  foci. 

2  =  Foci  are  moce  numerous  and  have  a  random 

distribution.  At  least  one  focus  per  20X 
microscopic  field  is  present,  but  is  restricted 
to  one  liver  lobe . 

3  =  At  least  one  focus  per  20X  field  is  present  in 

more  than  one  liver  lobe. 

4  =  Large  numbers  of  foci  (one  per  40X  field)  are 

present  in  one  or  more  hepatic  lobes. 

18.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  comments  and 
description  under  "General  Diagnoses"  above. 

19.  HEPATOCELLULAR  CARCINOMA  -  Malignant  neoplasm  of 
hepatocytic  origin.  Diagnostic  criteria  for  hepatocellular 
carcinoma  are  well  documented  in  the  literature.  The 
criteria  followed  in  this  study  were  those  of  Maronpot  et 
al..  Toxicologic  Pathology,  14:263-273,  1986.  Criteria  of 
importance  include  cellular  atypia,  irregular  shape,  local 
invasiveness,  haphazardly  arranged  cells,  broad  sheets  of 
cells,  trabecular  patterns,  and  glandlike  formations. 

Vascular  invasion  or  metastases  are  not  essential  for  making 
the  diagnosis  of  carcinoma,  but  may  be  present.  Hemorrhage 
and  necrosis  may  be  present . 

20.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

KIPHBX 

1.  PROGRESSIVE  RENAL  DISEASE  -  A  progressive  complex  of 
degenerative  changes  affecting  all  anatomic  components  of  the 
renal  cortex  and  medulla.  Interstitial  changes  begin  as 
small  aggregates  of  mononuclear  inflammatory  cells  and 
progress  to  interstitial  fibrosis  and  focal  mineralization. 
Tubular  epithelial  cells  undergo  degenerative  changes  such  as 
cytoplasmic  vacuolization  and  accumulation  of  hyaline 
droplets.  Cells  may  become  flattened  due  to  accumulation  of 


1 


Brown  et  al. — 428 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

eosinophilic  fluid  in  the' lumen,  or  they  may  become 
basophilic  and  hyperplastic  during  regenerative  attempts. 
Tubular  basement  membranes  are  frequently  thickened. 
Ultimately,  both  tubular  dilatation  and  atrophy  occur. 
Glomerular  changes  consist  of  thickening  of  Bowman's  capsule, 
pericapsular  fibrosis,  hypercellularity  of  the  glomerular 
capillary  tuft,  and  thickening  of  the  mesangial  matrix. 

These  changes  progress  to  glomerular  sclerosis  and/or  atrophy 
with  cystic  dilatation  of  Bowman's  space.  Hyalinization  of 
arteriole  walls  and  mineralization  of  tubular  basement 
membranes  and  necrotic  tubular  epithelial  cells  may  be 
present  in  severe  stages  of  the  disease.  Severity  grade 
criteria : 

1  =  There  are  occasional  aggregates  of  lymphocytes 

near  small  vessels  and  between  tubules.  An 
occasional  tubule  may  be  dilated  and  contain  a 
hyaline  cast . 

2  =*  More  tubules  are  dilated  and  contain  casts. 

Tubules  have  degenerative  changes  and  basement 
membranes  are  thickened.  Glomeruli  may  be 
hypercellular .  Up  to  25%  of  the  renal  cortex 
is  affected. 

3  =  Numerous  tubules  are  dilated  and  contain 

hyaline  material.  Tubular  regeneration  may  be 
present.  Interstitial  and  periglomerular 
fibrosis  is  present.  Glomerular  tufts  are 
thickened  and  hypercellular  and  a  few  glomeruli 
are  atrophied  and  dilated.  Renal  capsular 
surface  is  irregular.  Up  to  50%  of  the  kidney 
is  involved. 

4  =  Tubular  dilatation,  casts,  necrosis,  and 

mineralization  are  widespread.  Marked 
interstitial  fibrosis,  glomerular  atrophy  and 
dilatation,  and  irregularities  of  the  capsular 
surface  are  present.  There  is  hyalinization  of 
small  vessel  walls.  More  than  50%  of  the 
kidney  is  involved. 

2.  SARCOMA,  NOT  OTHERWISE  SPECIFIED  -  Presence  of  a  malignant 
mesenchymal  tumor  within  the  kidney.  Only  one  such  lesion 
was  detected  in  this  study  in  an  animal  that  did  not  have  a 
sarcoma  that  developed  within  the  fibrous  reaction  to  the 
test  material.  The  animal  having  this  lesion  is  a  male  in 
the  control  group.  This  lesion  is  a  well-differentiated 
fibrosarcoma.  Since  no  other  lesion  of  this  type  can  be 
found  in  this  animal,  it  is  assumed  that  the  kidney  is  the 


Brown  et  al. — 429 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 


primary  site. 

3.  HEMORRHAGE,  ACUTE  -  Presence  of  extravasated  blood  within 
the  kidney.  Probably  this  lesion  is  induced  during  necropsy 
procedures.  The  lesion  is  coded  only  as  "Present." 

4.  HEMOSIDEROSIS  (BROWN  GRANULAR  PIGMENT)  IN  TUBULAR 
EPITHELIAL  CELLS  -  Self-explanatory.  This  lesion  is  coded  as 
"Present . " 

5.  CYST (S) ,  CORTICAL  -  Presence  of  one  or  more  cystic  spaces 
within  the  renal  cortex.  Cysts  are  lined  by  flattened 
epithelium  and  may  be  either  empty  or  filled  with  pale 
homogeneous  eosinophilic  fluid.  While  this  may  be  a 
manifestation  of  progressive  renal  disease,  this  lesion  is 
coded  separately  if  the  cysts  are  more  than  three  times  the 
diameter  of  a  renal  tubule.  The  lesion  is  coded  as 
"Present . " 

6.  INFARCT,  CHRONIC  WITH  FIBROSIS,  WITH  OR  WITHOUT 
MINERALIZATION  -  Presence  of  well-demarcated,  usually  wedge- 
shaped  areas  in  the  cortex  composed  primarily  of  fibrous 
connective  tissue  and  a  few  pigment-laden  macrophages.  A  few 
degenerating  or  necrotic  tubular  epithelial  cells  and  mineral 
deposits  may  be  present.  At  the  periphery  of  the  lesion 
there  may  be  an  infiltration  of  mononuclear  inflammatory 
cells  and  occasional  areas  of  tubular  regeneration.  The 
lesion  is  coded  as  "Present." 

7.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
description  and  comments  under  "General  Diagnoses"  above. 

8.  FIBROSIS,  CAPSULE  -  Proliferation  of  non-neoplastic 
fibrous  connective  tissue  with  minimal  infiltration  of 
mononuclear  inflammatory  cells  in  the  renal  capsule.  The 
lesion  is  coded  as  "Present." 

9.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  description  and 
comments  under  "General  Diagnoses"  above. 

10.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  description  and 
comments  under  "General  Diagnoses"  above. 


PRINARY  BLADDER 

1.  TRANSITIONAL  CELL  PAPILLOMA  -  The  presence  of  a  neoplastic 


Brown  et  al. — 430 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

proliferation  of  transitional  epithelium.  The  cells 
comprising  this  tumor  are  well-differentiated  transitional 
epithelial  cells.  They  are  arranged  in  anastomosing  fronds 
which  protrude  into  the  lumen  of  the  urinary  bladder. 

Mitotic  figures  are  not  very  numerous.  An  occasional  tumor 
cell  is  undergoing  coagulative  necrosis.  The  tumor  is  highly 
vascularized.  There  is  no  evidence  of  neoplastic 
infiltration  into  the  submucosa  of  the  bladder  nor  invasion 
of  vessels  or  lymphatics. 

2.  CHRONIC  INFLAMMATORY  CELL  INFILTRATION  -  Presence  of 
lymphocytes,  macrophages,  and  plasma  cells  within  the  mucosa 
and  submucosa  of  the  urinary  bladder.  .  Severity  grade 
criteria : 

1  *  Minimal  detectable  inflammatory  cell 

infiltration  into  the  submucosa. 

2  =  Mild  inflammatory  cell  infiltration  which  may 

involve  both  mucosa  and  submucosa.  Less  than 
25%  of  the  circumference  of  the  bladder  is 
involved. 

3  -  Moderate  infiltration  of  the  mucosa  and 

submucosa  causing  thickening  of  these  layers 
and/or  involving  25%  to  50%  of  the 
circumference  of  the  bladder. 

4  =*  Marked  thickening  of  mucosa  and  submucosa  by 

inflammatory  cells  involving  more  than  50%  of 
the  circumference  of  the  bladder. 

PROSTATE 

1.  ACUTE  SUPPURATIVE  PROSTATITIS  -  Acute  infiltration  of 
neutrophils  into  glandular  or  interstitial  tissues  of  the 
prostate.  Necrosis  may  be  present  in  more  severe  cases. 
Severity  grade  criteria: 

1  -  Minimal  detectable  suppurative  inflammation 

involving  less  than  10%  if  the  prostate. 

2  -  Neutrophilic  infiltration  involving  10%  to  25% 

of  the  prostate. 

3  *  Neutrophilic  infiltration  involving  25%  to  50% 

of  the  prostate.  Small  areas  of  necrosis  may 
be  present. 

4  *  Infiltration  of  neutrophils  involves  more  than 

50%  of  the  prostate.  Necrotic  areas  up  to  the 
diameter  of  a  20X  microscopic  field  may  be 


Brown  et  al. — 431 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

present . 

2.  CHRONIC  PROSTATITIS  -  Infiltration  of  lymphocytes, 
macrophages,  plasma  cells,  and  a  few  neutrophils  into 
glandular  or  interstitial  areas  of  the  prostate.  The 
secretory  product  is  frequently  inspissated  in  affected  areas 
and  appears  as  small  eosinophilic  or  basophilic  particles 
intermixed  with  inflammatory  cells.  Mineralization  may  be 
present.  This  lesion  frequently  coexists  with  hyperplastic 
changes  in  the  glandular  epithelium  but  the  hyperplasia  is 
coded  separately  since  either  lesion  can  occur  independently. 
Severity  grade  criteria: 

1  =  Minimal  detectable  inflammatory  cell 

infiltration . 

2  =  Inflammatory  cell  infiltration  involves  10%  to 

25%  of  the  prostate. 

3  *»  Inflammatory  cell  infiltration  involves  25%  to 

50%  of  the  prostate. 

4  =  Inflammatory  cell  infiltration  involves  more 

than  50%  of  the  prostate.. 

3.  BENIGN  HYPERPLASIA  -  Hyperplastic  proliferation  of  the 
glandular  epithelium  of  the  prostate.  Secretory  cells  are 
larger  and  more  numerous  than  normal .  They  may  be  piled  two 
to  three  cell  layers  thick  on  the  basement  membrane.  There 
is  no  cellular  atypia  and  cells  are  distributed  in  normal 
anatomic  arrangement.  This  change  may  exist  with,  or 
independent  of  inflammatory  lesions  and  thus  is  coded 
separately.  Severity  grade  criteria: 

1  =  Hyperpl  3tic  changes  involve  up  to  10%  of  the 

glandular  epithelium. 

2  =  Hyperplastic  changes  involve  10%  to  25%  of  the 

epithelium. 

3  *  Hyperplastic  changes  involve  25%  to  50%  of  the 

epithelium. 

4  =  Hyperplastic  changes  involve  more  than  50%  of 

the  epithelium. 

4.  MICROCALICULI,  PROSTATIC  URETHRA  -  Presence  of  particulate 
material  in  the  prostatic  urethra.  The  material  consists  of 
an  amalgam  of  proteinaceous  or  mineralized  material  that  has 
variable  staining  characteristics  presumably  due  to 
variations  in  mineral  content.  There  may  be  mild 
infiltrations  of  mononuclear  inflammatory  cells  within  and 
beneath  the  urethral  epithelium.  The  lesion  is  coded  as 


Brown  et  al . — 432 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 


"Present . " 

5.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

COAGULATING  GLAND 

No  lesions  were  recognized  in  the  coagulating  glands  of 
rats  used  in  this  study. 

SEMINAL  VESICLE 

1.  GENERALIZED  ATROPHY  -  Presence  of  atrophic  changes  in  the 
glandular  epithelium  of  the  seminal  vesicle.  Epithelial 
cells  in  affected  glands  are  low  columnar  to  cuboidal, 
smaller  than  in  normal  actively  secreting  glands,  and  the 
meager  cytoplasm  stains  weakly  eosinophilic.  Glandular 
lumens  contain  very  little  secretory  product  in  contrast  to 
normal  glands  which  are  distended  with  this  material.  This 
change  is  coded  as  "Present." 

2.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

EPIDIDYMIS 

1.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

i&ax&a 

1.  TUBULAR  ATROPHY  -  The  cells  lining  seminiferous  tubules 
are  reduced  in  number,  and  the  tubules  are  reduced  in 
diameter.  This  change  is  invariably  present  in  cases  of 
interstitial  cell  adenoma  of  the  testis.  Consequently,  this 
lesion  is  not  coded  when  an  interstitial  cell  tumor  is 
present,  since  the  presence  of  the  tumor  itself  indicates 
that  tubular  atrophy  is  present.  Tubular  atrophy  is  coded 
when  testicular  neoplasia  was  not  present.  Severity  grade 
criteria  for  tubular  atrophy  are  as  follows: 

1  -  There  is  absence  or  reduction  in  the  number  of 

spermatozoa  or  spermatids  in  seminiferous 
tubules . 

2  -  In  addition  to  criteria  in  grade  1,  there  is 

reduction  in  the  innermost  layers  of 
spermatogenic  cells.  A  few  large 
multinucleated  cells  (degenerating 


Brown  et  al. — 433 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

spermatocytes)  may  be  free  in  the  tubular 
lumens . 

3  *  Only  a  few  spermatogenic  cells  are  present 

along  with  a  complete  layer  of  Sertoli  cells  on 
the  basement  membrane.  Multinucleated  cells 
may  be  present. 

4  *  There  is  complete  absence  of  spermatogenic 

cells.  Seminiferous  tubules  contain  only 
Sertoli  cells. 

2.  INTERSTITIAL  CELL  HYPERPLASIA  -  There  is  focal  or 
multifocal  proliferation  of  interstitial  cells.  These  are 
large  cells  with  round  nuclei  and  abundant  eosinophilic 
cytoplasm  which  may  be  finely  vacuolated.  They  are  situated 
between  seminiferous  tubules.  The  change  is  considered 
hyperplasia  when  foci  of  interstitial  cells  are  no  larger 
than  the  diameter  of  a  seminiferous  tubule  and  there  is  no 
compression  of  adjacent  tissue.  Foci  of  interstitial  cells 
larger  than  the  diameter  of  a  tubule,  or  that  compress 
adjacent  tissue  are  coded  as  interstitial  adenomas.  If  both 
adenoma  and  hyperplasia  is  present,  only  the  adenoma  is 
coded.  Severity  grade  criteria: 

1  *  One  to  three  foci  of  hyperplastic  interstitial 

cells  are  present. 

2  -  Four  to  six  foci  are  present . 

3  *  Seven  to  ten  foci  are  present . 

4  =  More  than  ten  foci  are  present . 

3.  MESOTHELIOMA  -  Neoplastic  proliferation  of  mesothelial 
cells,  arranged  in  multiple  papillary  fronds,  arising  from 
the  tunica  albuginea  of  the  testis.  The  papillary  structures 
are  supported  by  a  connective  tissue  stalk  that  contains 
blood  vessels.  The  neoplastic  mesothelial  cells  may  be 
arranged  in  a  single  layer  of  flattened  cells,  or  the  cells 
may  be  cuboidal  and  piled  up  to  form  small  nodules.  Mitotic 
figures  are  usually  infrequent. 

4.  INTERSTITIAL  CELL  ADENOMA  -  A  benign  tumor  of  interstitial 
(Leydig)  cells.  These  tumors  are  composed  of  cells  of  four 
distinct  morphologic  types.  The  most  common  is  a  medium¬ 
sized  hexagonal  cell  with  distinct  cell  boundaries  having 
eosinophilic  cytoplasm  surrounding  a  regular,  round  or  oval 
vesicular  nucleus.  The  nucleus  has  a  delicate  chromatin 
network  and  a  single  small  basophilic  nucleolus.  The  second 
cell  type  is  larger  with  finely  or  coarsely  vacuolated 
cytoplasm.  A  third  cell  type  is  small,  elongated  or  spindle 


Brown  et  al . — 434 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

shaped,  with  granular  eosinophilic  cytoplasm  and  variably- 
sized,  round,  oval,  or  spindle-shaped  nuclei  which  may  be 
vesicular  or  pyknotic.  The  fourth  type  of  cell  resembles  a 
lymphocyte  with  scanty  cytoplasm  and  a  small  round  nucleus 
with  a  "wheel-like"  chromatin  pattern.  Neoplastic  cells  are 
arranged  in  cords,  fascicles,  islands,  and  nests,  and  usually 
show  an  endocrine  pattern  of  vascularity.  The  stroma  is 
usually  delicate,  but  may  be  hyalinized  or  even  calcified. 
These  tumors  are  highly  vascular  and  often  there  are  large, 
distended  endothelial-lined  spaces  filled  with  blood. 
Invariably,  there  are  secondary  lesions  in  the  testis  when 
interstitial  cell  adenoma  is  present.  Atrophic  or 
degenerative  changes  affecting  seminiferous  tubules  are 
almost  always  present,  and  hemorrhage,  necrosis,  and 
mineralization  may  occur  within  the  tumor.  These  lesions  are 
not  coded  separately  when  interstitial  adenoma  is  present. 

5.  PERIARTERITIS  NODOSA  -  An  acute,  subacute,  or  chronic 
inflammatory  lesion  involving  small  or  medium-sized  muscular 
arteries  in  the  testis.  This  lesion  is  characterized  by  sub- 
endothelial  edema,  damage  to  the  elastic  membrane,  fibrinoid 
or  hyaline  necrosis  of  the  media,  and  inflammatory  cell 
infiltration  into  all  layers  of  the  arterial  wall. 
Inflammatory  cells  are  most  abundant  in  the  adventitia  and 
are  composed  of  neutrophils,  lymphocytes,  and  plasma  cells 
with  fewer  macrophages  and  eosinophils .  It  occurs  as  a  focal 
or  segmental  lesion  with  some  areas  of  an  artery  severely 
affected,  and  other  areas  essentially  normal.  No  attempt  is 
made  to  grade  the  severity  of  this  lesion.  It  is  coded  as 
"Present . " 

6.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

7.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  descriptions 
and  comments  under  "General  Diagnoses"  above. 

2.  DILATATION  OF  THE  UTERINE  LUMEN  (HYDROMETRA)  -  There  is 
dilatation  of  the  uterine  lumen  which  is  usually  empty.  The 
endometrium  and  underlying  stroma  may  be  slightly  atrophic. 
The  lesion  is  coded  as  "Present." 


Brown  et  al. — 435 


Appendix  I  (cont.) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

3.  PYOMETRA  -  There  is  suppurative  inflammation  involving  the 
uterine  lumen,  endometrium,  endometrial  glands,  and  stroma. 
Small  foci  or  areas  of  necrosis  may  be  present  in  areas 
severely  affected.  Severity  grade  criteria: 

1  =  Presence  of  a  few  neutrophils  within  the 

endometrium. 

2  =  Moderate  numbers  of  neutrophils  are  present 

within  the  endometrium,  endometrial  glands,  and 
stroma . 

3  =  Neutrophilic  infiltration  as  in  grade  2  and 

presence  of  neutrophils  within  the  uterine 
lumen . 

4  =  Changes  as  in  grade  3  plus  distention  of  the 

uterine  lumen  with  suppurative  exudate  and/or 
the  extension  of  inflammatory  process  into  the 
uterine  musculature. 

4.  ENDOMETRIAL  STROMAL  POLYP  -  These  lesions  consist  of 
polypoid  masses  which  protrude  from  the  uterine  wall  into  the 
lumen.  The  mass  is  covered  by  essentially  normal  endometrial 
surface  epithelium.  Within  the  mass  there  are  relatively 
normal-appearing  uterine  glands;  some  may  be  dilated  and 
cystic.  The  mass  consists  primarily  of  dense  or  loosely 
arranged  f ibrovascular  stroma  containing  collagen  and 
numerous  small  vessels .  Some  smooth  muscle  cells  may  be 
present.  A  few  mitotic  figures  may  be  present  in  both  stroma 
and  epithelium.  Focal  ulceration  of  the  epithelium 
accompanied  by  inflammatory  cell  infiltration  may  be  present. 
The  lesion  is  coded  as  "Present." 

5.  CYSTIC  DILATATION  OF  ENDOMETRIAL  GLANDS  -  Self- 
explanatory.  Coded  as  "Present." 

6.  LYMPHOCYTIC  INFILTRATE  -  Presence  of  lymphocytes  within 
the  endometrium,  stroma,  or  uterine  wall.  Severity  grade 
criteria: 

1  -  There  are  a  few  lymphocytes  within  the 

endometrium. 

2  -  There  is  mild  lymphocytic  infiltrate  in  the 

endometrium  and  around  uterine  glands. 

3  -  There  are  moderate  numbers  of  lymphocytes  in 

the  endometrium  and  endometrial  stroma. 

4  *  The  lymphocytic  infiltrate  involves 

endometrium,  stroma,  and  extends  into  the 
uterine  musculature. 


Brown  et  al . — 436 


Appendix  I  (cont. ) :  GLOSSARY  Or  TERMS 

FOR  PATHOLOGY  FINDINGS 

7 .  ENDOMETRIAL  STROMAL  SARCOMA  -  A  malignant  proliferation  of 
endometrial  stromal  cells.  These  are  usually  highly  cellular 
masses  which  may  protrude  into  the  uterine  lumen  or  invade 
the  uterine  musculature.  Cells  are  usually  spindeloid  with 
elongated  dense  hyper chromatic  nuclei  and  scanty  amounts  of 
cytoplasm  which  are  either  weakly  eosinophilic  or  nearly 
colorless.  Cells  are  arranged  in  a  variety  of  patterns 
including  parallel  arrays,  interlacing  bundles,  or  haphazard 
proliferations.  Cells  often  are  fairly  anaplastic  and 
mitotic  figures  may  be  numerous.  Hemorrhage  and  necrosis  are 
common  especially  in  the  large  polypoid  masses.  The  surface 
epithelium  and  endometrial  glands  are  usually  normal, 
although  inflammation  and  degeneration  of  these  components 
may  occur.  These  tumors  differ  from  stromal  polyps  in  their 
increased  cellularity,  cellular  atypia,  and  invasiveness.  No 
metastatic  lesions  of  this  type  were  observed  in  this  study. 

8.  ENDOMETRITIS,  SUBACUTE  -  Subacute  inflammatory  reaction 
involving  the  endometrium.  There  is  infiltration  of 
neutrophils,  lymphocytes,  macrophages,  and  plasma  cells  into 
the  endometrial  epithelium,  underlying  stroma,  and  uterine 
glands.  This  lesion  differs  from  pyometra  (Diagnosis  #3)  in 
that  the  inflammatory  cell  population  is  mixed  and  no 
suppurative  exudate  is  present  in  the  uterine  lumen.  The 
mixed  cell  population  differentiates  this  lesion  from  the 
pure  lymphocytic  infiltrate  described  in  Diagnosis  #6  above. 
Severity  grade  criteria  for  this  lesion: 

1  -  There  is  minimal  infiltration  of  mixed 

inflammatory  cells  in  the  endometrium. 

2  =  There  is  mild  infiltration  of  inflammatory 

cells  in  endometrium,  and  into  and  around 
uterine  glands . 

3  *  There  is  moderated  infiltration  of  inflammatory 

cells  in  endometrium  and  all  areas  of  the 
stroma.  Minimal  amounts  of  necrosis  may  be 
present  in  these  areas . 

4  -  There  are  marked  mixed  inflammatory  cell 

infiltrate  and  areas  of  necrosis  that  involve 
endometrium,  stroma,  and  uterine  musculature. 

9.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
descriptions  and  comments  under  "General  Diagnoses"  above. 

10.  DEGENERATION,  NECROSIS  AND  INFLAMMATION  OF  UTERINE  STROMA 
-  This  unusual  lesion  is  characterized  by  t  abundance  of 
necrotic  debris  and  fibrin  within  the  uterine  lumen. 


Brown  et  al. — 437 


Appendix  I  (coat.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

degeneration,  necrosis,  and  proliferation  of  endometrial 
stromal  cells,  dilatation  of  endometrial  glands,  moderate 
mixed  inflammatory  cell  infiltration  into  all  layers  of  the 
uterine  wall,  and  the  presence  of  many  large  cells  with 
abundant  poorly  stained  cytoplasm.  These  cells  are  situated 
primarily  on  the  lumenal  surface  of  the  uterus  but  in  some 
areas  extend  through  the  stroma  and  infiltrate  the  uterine 
musculature.  Sections  stained  with  periodic  acid-Schiff, 
alcian  blue,  and  mucicarmine  stains  failed  to  characterize 
the  cytoplasmic  content  of  these  cells.  Their  identity  and 
character  remain  unknown.  This  unusual  lesion  was  observed 
in  only  one  rat,  a  female  in  the  Control  group  that  survived 
until  terminal  sacrifice.  The  morphologic  characteristics  of 
this  lesion  do  not  correspond  to  the  criteria  of  the  other 
more  readily  recognized  uterine  lesions  observed  in  this 
study,  hence  this  descriptive  diagnosis  is  given. 

11.  LEIOMYOSARCOMA  -  A  malignant  neoplasm  of  smooth  muscle 
origin.  This  is  a  highly  cellular  tumor  characterized  by 
spindeloid  cells  arranged  in  interlacing  bundles,  and 
situated  in  the  wall  of  the  uterus.  The  cells  are  large  and 
spindeloid  to  fusiform  with  moderate  amounts  of  eosinophilic 
cytoplasm  and  elongated  nuclei  with  blunt  or  rounded  ends. 

The  cells  are  well  differentiated  and  mitotic  figures  are  not 
numerous.  The  tumor  infiltrates  and  separates  adjacent 
normal  smooth  muscle  cells  in  an  irregular  fashion  and 
invades  the  adjacent  endometrial  stroma. 

OVARIES 

No  attempt  is  made  to  document  ovarian  changes  that 
occur  as  a  result  of  different  stages  of  the  estrus  cycle,  or 
that  result  from  normal  variations  in  hormonal  stimulation. 
Only  changes  considered  abnormal  are  documented  in  this 
study . 

1.  FOLLICULAR  CYSTS  -  Presence  of  large  cystic  structures  in 
the  ovarian  cortex.  Cysts  are  lined  by  a  layer  of  granulosal 
cells  which  varies  considerably  in  thickness.  Degeneration 
and  atrophy  of  the  lining  cell  layers  are  common  and  there 
may  be  a  few  degenerating  cells  free  within  the  pale 
eosinophilic  fluid  that  fills  the  cystic  lumen.  The  cysts 
are  coded  as  "Present." 

2.  CYSTIC  CORPORA  LUTEA  -  Presence  of  cystic  structures  in 
the  ovarian  cortex.  Cysts  are  lined  by  a  layer  of  luteal 


Brown  et  al . — 438 


Appendix  I  (cont.):  GLOSSARY  OP  TERMS 

TOR  PATHOLOGY  FINDINGS 

cells  that  varies  considerably  in  thickness.  Degeneration, 
atrophy,  and  infiltration  of  a  very  few  mononuclear 
inflammatory  cells  may  occur.  The  lumen  of  the  cyst  may  be 
empty  or  filled  with  pale  eosinophilic  fluid.  The  cysts  are 
coded  as  "Present.” 

3.  PARA-OVARIAN  CYST(S)  -  Presence  of  a  cystic  structure 
located  adjacent  to  the  ovary  but  does  not  involve  ovarian 
tissue.  Frequently  this  lesion  consists  only  of  a  thin 
fibrous  strand  of  tissue  (the  wall  of  the  cyst)  which  may  be 
lined  by  a  single  layer  of  flat  "endothelial-like"  cells. 
Ovaries  adjacent  to  these  cysts  may  be  small  and  atrophic. 
Para-ovarian  cysts  are  coded  as  "Present." 

4.  INFLAMMATORY  CELL  INFILTRATION  -  The  presence  of 
lymphocytes,  macrophages,  plasma  cells,  or  (rarely) 
neutrophils  within  ovarian  tissues.  This  lesion  is  observed 
infrequently.  Severity  grade  criteria  are  subjective 
assessments  of  the  number  of  inflammatory  cells  present: 

1  =  Minimal  inflammatory  cells  present. 

2  *  Mild  inflammatory  cell  infiltration. 

3  =  Moderate  inflammatory  cell  infiltration. 

4  -  Marked  inflammatory  cell  infiltration. 

5.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
descriptions  and  comments  under  "General  Diagnoses"  above. 

6.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

QUQPBHPM 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  description 
and  comments  under  "General  Diagnoses"  above. 

2.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

3.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  descriptions  and 
comments  under  "General  Diagnoses"  above. 

4.  MUCOSAL  EROSION/ULCERATION  -  Necrosis  and  sloughing  of  the 
epithelium  on  the  mucosal  surface.  In  an  erosion,  the 
necrosis  is  limited  to  the  epithelial  layer;  the  basement 
membrane  is  intact.  An  ulceration  consists  of  necrosis  that 


Brown  et  ai . — 439 


Appendix  I  (cont . ) :  GLOSSARY  OT  TERMS 

FOR  PATHOLOGY  FINDINGS 

involves  the  surface  epithelium,  extends  through  the  basement 
membrane,  and  affects  the  underlying  submucosa.  In  both 
lesions,  tissues  adjacent  to  necrotic  areas  are  infiltrated 
with  neutrophils,  lymphocytes,  and  macrophages.  In  chronic 
ulcers,  there  may  be  fibrous  connective  tissue  proliferation 
and  regenerating  mucosal  epithelium  at  the  periphery  of  the 
lesion.  These  lesions  are  coded  as  "Present." 

5.  PRESENCE  OF  MUCOSAL  EPITHELIAL  CELLS  WITHIN  INTESTINAL 
WALL  AND  ADJACENT  FIBROUS/GRANULOMATOUS  INFLAMMATORY  REACTION 
-  In  some  animals  the  test  material  adhered  to  the  intestinal 
wall  inciting  the  usual  foreign-body  inflammatory  response. 

In  some  of  these  animals,  there  is  chronic  ulceration  of  the 
epithelium  at  this  location.  Microscopic  examination  reveals 
the  presence  of  regenerating  intestinal  epithelial  cells  in 
the  submucosa,  within  the  muscular  layers,  in  subserosal 
locations,  and  within  the  adjacent  chronic  granulomatous 
inflammatory  tissue.  These  entrapped  cells  have  essentially 
normal  histologic  characteristics  except  for  their  location 
and  occasional  degenerative  or  atrophic  changes.  There  are 
no  indications  of  cellular  atypia,  nuclear  pleomorphism,  or 
abnormal  mitotic  activity  characteristic  of  neoplasia.  This 
change  is  interpreted  as  regeneration  of  intestinal 
epithelium  which  has  migrated  into,  and  become  entrapped  in 
an  ongoing  chronic  inflammatory  response  that  affects  the 
entire  thickness  of  the  intestinal  wall.  This  lesion  is 
coded  as  "Present." 

JBJPMPM 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  "General 
Diagnoses"  above. 

2.  ACUTE  PERITONITIS  -  The  presence  of  neutrophils,  fibrin, 
necrotic  debris,  and  mononuclear  inflammatory  cells  on  the 
serosal  surface.  No  test  material,  foreign-body  giant  cells, 
or  fibrosis  are  present.  Severity  grade  is  based  on 
subjective  assessment  of  the  intensity  and  extent  of  the 
inflammatory  process : 

1  -  Minimal  inflammation. 

2  ■  Mild  inflammation. 

3  -  Moderate  inflammation. 

4  =  Marked  inflammation. 


3.  ENTERITIS,  SUBACUTE  -  Consists  of  the  infiltration  of 


Brown  et  al. — 440 


Appendix  I  (cont . ) :  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

lymphocytes,  macrophages,  plasma  cells,  and  a  few  neutrophils 
into  the  mucosa,  lamina  propria,  submucosa,  and  muscular  wall 
of  the  jejunum.  In  more  severe  cases  there  may  be  small 
areas  of  necrosis  in  the  layers  involved.  Severity  grade 
criteria : 

1  =  Minimal  inflammatory  cell  infiltration  in 

mucosa  and/or  lamina  propria. 

2  =  Mild  inflammatory  cell  infiltration  with  edema, 

in  mucosa,  lamina  propria,  and  extending  into 
the  submucosa. 

3  =  A  moderate  inflammatory  infiltrate  and  edema 

involves  mucosa,  lamina  propria  and  submucosa 
causing  increased  thickness  of  these  layers. 
Small  foci  or  areas  of  necrosis  may  be  present 
in  the  mucosa. 

4  =  A  marked  inflammatory  cell  infiltrate 

accompanied  by  edema,  involves  mucosa,  lamina 
propria,  submucosa,  and  extends  into  the 
muscular  coats.  Foci  or  areas  of  necrosis  are 
present  in  the  mucosa  and  lamina  propria  and 
may  involve  deeper  layers . 

4.  MUCOSAL  EROSION/ULCERATION  -  Except  for  location  in  the 
jejunum,  this  lesion  is  similar  to  that  described  in 
Diagnosis  #4  under  DUODENUM  above. 

5.  PRESENCE  OF  MUCOSAL  EPITHELIAL  CELLS  WITHIN  INTESTINAL 
WALL  AND  ADJACENT  FIBROUS /GRANULOMATOUS  INFLAMMATORY  REACTION 
-  Except  for  location  in  the  jejunum  this  lesion  is  similar 
to  that  described  ir.  Diagnosis  #5  under  DUODENUM  above. 

6.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  "General  Diagnoses" 
above . 

7.  FIBROVASCULAR  ADHESION,  SEROSA  -  Self-explanatory. 

Minimal  inflammatory  cell  infiltration,  no  test  material  and 
no  foreign-body  giant  cells  are  present.  Lesion  is  coded  as 
"Present . " 

8.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  "General  Diagnoses" 
above . 

ILEUM 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  "General 


Brown  et  al . —  441 


Appendix  I  (cont . ) :  GLOSSARY  or  TERMS 

FOR  PATHOLOGY  FINDINGS 


Diagnoses"  above. 

2.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
"General  Diagnoses"  above. 

3.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  "General  Diagnoses" 
above . 

4.  LYMPHOID  HYPERPLASIA  -  This  change  involves  the  gut- 
associated  lymphoid  tissue  in  the  ileum,  and  consists  of 
increased  size  and  number  of  lymphoid  follicles,  increased 
number  of  large,  pale,  immature  lymphocytes  in  germinal 
centers  and  perifollicular  areas,  increased  mitotic  activity 
in  lymphoid  cells,  and  the  extension  of  lymphoid  cells  from 
the  submucosa  into  the  lamina  propria  and  mucosal  layers  of 
the  ileum.  This  change  is  coded  as  "Present." 

5.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  "General  Diagnoses" 
above . 

FAMgRfifta 

1.  ACINAR  (EXOCRINE)  ATROPHY  -  This  lesion  consists  of  the 
absence  or  reduction  in  number  of  acinar  secretory  cells. 
Affected  lobules  contain  only  ductal  structures  in  a 

f ibrovascular  stroma.  Such  lesions  are  usually  focal 
affecting  a  small  lobule  of  pancreatic  tissue.  Severity 
grade  criteria: 

1  =  One  lobule  affected. 

2  =  Two  or  three  lobules  affected. 

3  =  Four  to  six  lobules  affected. 

4  =  More  than  six  lobules  affected. 

2.  CHRONIC  INFLAMMATION  -  Infiltration  of  neutrophils 
lymphocytes,  macrophages,  and  plasma  cells  into  acinar  areas, 
stromal  tissues,  and  serosal  surfaces  of  the  pancreas.  There 
may  be  variable  amounts  of  edema,  fibrin  deposition, 
fibrosis,  tissue  necrosis,  and  mineralized  deposits  in 
necrotic  areas  depending  on  the  severity  and  age  of  the 
lesion.  Severity  grade  criteria: 

1  =  Minimal  focal  to  multifocal  inflammatory  cell 

infiltration . 

2  =  Mild  focal  to  diffuse  inflammatory  cell 

infiltrate  with  or  without  fibrosis,  affecting 


Brown  et  al . — 442 


Appendix  I  (cont.):  GLOSSARY  or  TERMS 

FOR  PATHOLOGY  FINDINGS 

up  to  50%  of  the  pancreas . 

3  =  A  few  small  foci  of  necrosis  may  be  present 

along  with  moderate  inflammatory  cell 
infiltration  and  fibrosis. 

4  =  Numerous  foci  or  large  areas  of  necrosis  which 

may  be  accompanied  by  mineralization. 
Inflammatory  cell  infiltrate  and  fibrosis  are 
marked  and  diffuse. 

3.  ISLET  CELL  ADENOMA  -  Benign  neoplastic  proliferation  of 
pancreatic  islet  cells.  These  adenomas  are  usually  nearly 
spherical  proliferations  of  medium-sized,  well-differentiated 
islet  cells.  The  cells  are  arranged  in  small  nests,  packets, 
or  short  cords  supported  by  very  delicate  f ibrovascular 
stroma.  Cytoplasm  is  weakly  eosinophilic  and  has  distinct 
cellular  boundaries.  Nuclei  are  round  and  fairly  small  with 
condensed  chromatin.  Mitotic  figures  are  sparse.  The  tumors 
are  discrete  and  compress  adjacent  tissues  slightly. 

4.  PERIARTERITIS  NODOSA  -  An  acute,  subacute,  or  chronic 
inflammatory  lesion  involving  small  to  medium-sized  muscular 
arteries  in  the  pancreas.  The  lesion  is  characterized  by 
subendothelial  edema,  hyaline,  or  fibrinoid  necrosis  of  the 
media,  and  infiltration  of  all  layers  of  the  arterial  wall  by 
inflammatory  cells.  Inflammatory  cell  infiltration  is  most 
prevalent  in  the  adventitia  and  consists  of  neutrophils, 
lymphocytes,  and  plasma  cells  with  fewer  macrophages  and 
eosinophils.  The  lesion  is  usually  focal  or  segmental  with 
some  areas  of  an  artery  being  severely  affected  and  other 
areas  essentially  normal.  The  lesion  is  coded  as  "Present." 

5.  ACINAR  HYPERPLASIA  -  Focal  or  multifocal  non-neoplastic 
proliferation  of  pancreatic  acinar  secretory  cells.  Within 
an  affected  lobule,  there  are  increased  numbers  of  secretory 
cells  which  are  either  normal  size  or  slightly  smaller  than 
normal.  The  cells  are  crowded,  but  are  arranged  in 
essentially  normal  fashion  relative  to  basement  membranes. 
Hyperplastic  cells  are  usually  slightly  hyperchromatic  but 
often  lack  normal  numbers  of  zymogen  granules.  Severity 
grade  criteria: 

1  -  One  pancreatic  lobule  is  affected. 

2  *  Two  or  three  lobules  are  affected. 

3  =  Four  to  six  lobules  are  affected. 

4  =  More  than  six  lobules  are  affected. 


6.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  "General 


Brown  et  al. — 443 


Appendix  I  (cont.) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 


Diagnoses"  above. 

7.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  "General  Diagnoses” 
above . 

8.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA,  METASTATIC  -  See 
"General  Diagnoses"  above. 

9.  PRESENCE  OF  METASTATIC  NEOPLASM  -  See  "General  Diagnoses" 
above . 

10.  CYSTIC  DUCT  -  Cystic  dilatation  of  one  or  more  ducts 
within  the  pancreas.  Dilated  area  is  more  than  three  times 
the  diameter  of  a  normal  duct .  The  lumen  is  usually  empty 
and  the  lining  epithelium  may  be  flattened  and  atrophic.  The 
lesion  is  coded  as  "Present." 

CECUM 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  "General 
Diagnoses"  above. 

2.  PERIARTERITIS  NODOSA,  ARTERIES  IN  CECAL  WALL  OR  MESENTERY 
-  See  descriptions  and  comments  for  Diagnosis  #4  under 
PANCREAS  above.  The  characteristics  of  the  arterial  changes 
are  similar  in  both  organs. 

3.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  "General  Diagnoses" 
above . 

4.  ADENOMATOUS  POLYP  -  This  lesion  is  a  large  polypoid  mass 
attached  to  the  wall  of  the  cecum  by  a  broad  base.  The 
intralumenal  surface  of  the  mass  is  covered  by  well- 
differentiated  epithelium  arranged  in  long  tortuous  tubular 
glands.  The  lamina  propria  between  glands  is  slightly 
separated  by  edema  and  a  mild  uniform  infiltration  of 
lymphocytes  and  a  few  plasma  cells.  The  bulk  of  the  mass 
consists  of  the  underlying  stroma  which  is  characterized  by 
numerous  endothelial-lined  spaces  that  vary  considerably  in 
size  and  shape.  Most  of  the  vascular  spaces  are  small  and 
slit-like,  although  there  are  a  few  that  are  larger  and 
irregular  in  shape.  Most  of  these  spaces  are  empty.  Between 
the  vascular  spaces  there  are  numerous  loosely-arranged 
fibrocytes  and  small  foci  of  mononuclear  inflammatory  cells. 

A  few  small  muscular  arteries  are  present  in  this  area.  Only 
one  lesion  of  this  type  was  observed  in  this  study.  It 
occurred  in  a  female  rat  in  the  control  group. 


Brown  et  al. — 444 


Appendix  I  (cont . ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

5.  REACTIVE  LYMPHOID  HYPERPLASIA,  SUBMUCOSA  -  This  change  is 
essentially  identical  to  that  in  the  ILEUM.  See  diagnosis  #4 
under  ILEUM  above  for  a  detailed  description. 

6.  EDEMA  OF  THE  SUBMUCOSA  AND/OR  LAMINA  PROPRIA  -  The 
cellular  elements  and  connective  tissue  in  the  submucosa  and 
lamina  propria  are  separated  by  the  presence  of  weakly 
eosinophilic  edema  fluid.  This  lesion  is  coded  as  "Present." 

7.  MONONUCLEAR  CELL  INFILTRATION  -  The  lamina  propria  and 
submucosa  are  infiltrated  by  small  numbers  of  lymphocytes, 
plasma  cells,  and  macrophages.  The  lesion  is  coded  as 
"Present." 

RE C TOM 

1.  FIBROSARCOMA,  LOCALLY  INVASIVE  -  These  tumors  are  highly 
cellular  and  consist  of  spindeloid  cells  with  varying  amounts 
of  eosinophilic  cytoplasm  and  oval  to  elongated  nuclei. 
Mitotic  figures  may  be  very  numerous  and  large  bizarre  forms 
occur  occasionally.  The  cells  are  arranged  in  broad  sheets, 
parallel  arrays,  and  interlacing  bundles.  In  more  anaplastic 
tumors,  a  definite  cellular  pattern  may  be  difficult  to 
detect.  Sections  stained  with  Masson's  trichrome  stain 
usually  reveal  the  presence  of  abundant  collagen.  The  most 
probable  primary  site  for  this  tumor  is  the  inguinal  or 
perineal  subcutaneous  tissue.  Three  fibrosarcomas  were 
present  in  this  location  in  Study  GLP  83009.  All  three 
animals  were  females;  one  in  the  control  group,  one  in  the 
high-dose  group  and  one  in  the  low-dose  group.  In  all  three 
animals,  the  tumors  were  confined  to  the  inguinal  or  perineal 
subcutis  with  local  invasion  of  adjacent  pelvic  viscera.  In 
none  of  these  cases  was  there  evidence  of  the  sarcoma 
elsewhere  in  the  abdominal  cavity.  No  test  material  or 
granulomatous  inflammatory  lesions  were  observed  in 
association  with  these  perirectal  tumors. 

C.QJL&H 

1.  COMPOUND-PELATED  INFLAMMATORY  REACTION  -  See  "General 
Diagnoses"  above. 

2.  CHRONIC  COLITIS  WITH  MUCOUS  GLAND  HYPERPLASIA  AND 
HYPERSECRETION  -  The  mucosa,  lamina  propria,  and  submucosa 
are  infiltrated  with  lymphocytes,  macrophages,  plasma  cells, 
and  an  occasional  neutrophil.  Many  of  the  glands  in  the 


Brown  et  al. — 445 


Appendix  I  (cont . )  :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

mucosa  are  distended  with  mucus.  Small  amounts  of  mucus 
adhere  to  the  mucosal  surface  and  are  present  in  the  lumen. 
Epithelial  cells  in  the  mucosal  glands  are  hyperplastic. 
Severity  grade  criteria  are  not  applicable  since  only  two 
animals  in  this  study  had  this  lesion. 

3.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  "General  Diagnoses" 
above . 

4.  POLYP,  CYSTIC,  WITH  MUCINOUS  EPITHELIAL  HYPERPLASIA  AND 
OSSEOUS  METAPLASIA  AND/OR  MINERALIZATION  OF  THE  STROMA  - 
These  unusual  lesions  consist  of  large  polypoid  growths  which 
extend  into  the  lumen  of  the  colon.  The  mass  is  covered  by 
colonic  epithelium  which  in  some  areas  is  ulcerated  and 
accompanied  by  a  chronic  inflammatory  response  consisting  of 
mononuclear  cell  infiltration  and  fibrosis.  Within  the  mass 
are  numerous  large  cystic  spaces  that  are  lined  by  flattened 
epithelium  containing  numerous  mucus-secreting  cells.  These 
cystic  cavities  are  distended  with  mucus  that  contains  a  few 
inflammatory  cells  and  small  particles  of  cellular  debris. 

The  stroma  between  the  mucus-filled  cysts  consists  of 
numerous  slit-like  endothelial-lined  vascular  spaces,  fibrous 
connective  tissue  which  is  very  dense  in  some  areas  and 
loosely  arranged  and  edematous  in  others,  and  a  multifocal  to 
diffuse  infiltration  of  mononuclear  inflammatory  cells.  In 
some  areas  the  dense  fibrous  stroma  is  mineralized  and  in 
other  places  there  are  spicules  of  metaplastic  bone.  The 
adjacent  colonic  mucosa  is  ulcerated,  infiltrated  with 
inflammatory  cells,  and  contains  areas  of  fibrous  connective 
tissue  proliferation  and  foci  of  hyperplastic  (regenerating) 
epithelial  cells.  These  lesions  occur  in  only  two  animals; 
both  males  in  the  high-dose  group.  In  both  cases  there  are 
serosal  adhesions  of  the  colon  that  could  be  attributed  to 
the  presence  of  the  test  material.  In  one  case,  the  cecum 
and  its  cystic  polypoid  mass  are  adhered  to  the  surface  of 
the  liver.  It  is  probable  that  the  presence  of  the  test 
material  an'’,  the  reaction  to  it  contributed  to  the 
development  of  these  unusual  lesions  in  the  colon.  The 
lesion  is  coded  as  "Present." 

STQMAfia 

Mucosal  glandular  dilatation  was  observed 
microscopically  in  almost  every  rat  that  was  necropsied 
during  the  second  year  of  this  study.  Since  this  change  is 
universally  present  in  all  three  experimental  groups,  it  is 


Brown  et  al . — 446 


Appendix  I  (cont.) :  GLOSSARY  or  TERMS 

rOR  PATHOLOGY  FINDINGS 

considered  as  a  normal  aging  change  and  is  not  coded. 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  "General 
Diagnoses"  above. 

2.  SEROSAL  FIBROSIS  -  The  presence  of  fibrous  connective 
tissue  proliferation  on  the  serosal  surface.  Minimal  numbers 
of  inflammatory  cells  may  be  present.  There  is  no  test 
material  or  foreign-body  giant  cells.  The  lesion  is  coded  as 
"Present . " 

3.  ULCERATION,  NONGLANDULAR  AREA  -  There  is  necrosis  and 
sloughing  of  the  squamous  epithelial  lining  of  the  non- 
glandular  portion  of  the  stomach.  Mononuclear  inflammatory 
cell  infiltration,  edema,  and  proliferation  of  small  amounts 
of  fibrous  connective  tissue  may  be  present  at  the  periphery 
of  the  ulcer  and  in  underlying  stomach  wall.  The  lesion  is 
coded  as  "Present." 

4.  MUCOSAL  AND/OR  SUBMUCOSAL  EDEMA  -  Cellular  elements  of  the 
stomach  mucosa  and/or  submucosa  are  separated  by  the  presence 
of  pale  eosinophilic  edema  fluid.  This  lesion  may  occur  in 
either  the  glandular  or  squamous -lined  portions  of  the 
stomach.  The  lesion  is  coded  as  "Present." 

5.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  See  "General 
Diagnoses"  above. 

6.  PRESENCE  OF  METASTATIC  SARCOMA  -  See  "General  Diagnoses" 
above . 

7.  SQUAMOUS  CELL  PAPILLOMA  -  There  is  a  well-defined 
proliferation  of  the  squamous  epithelium  of  the  stomach.  The 
proliferating  epithelial  cells  are  well  differentiated  and 
are  arranged  in  blunt  papillary  projections  which  extend  into 
the  lumen.  The  surface  of  the  growth  is  usually  keratinized. 
Proliferation  of  basal  cells  can  be  seen.  The  growth  does 
not  extend  beyond  the  basement  membrane .  The  mucosa  and 
submucosa  adjacent  to  papillomas  are  usually  infiltrated  by 
mononuclear  inflammatory  cells. 

8.  EROSION  AND/OR  ULCERATION  OF  THE  GLANDULAR  MUCOSA  -  There 
is  necrosis  and  sloughing  of  the  epithelial  cells  in  the 
glandular  portion  of  the  stomach  mucosa.  In  an  erosion,  the 
necrosis  is  confined  to  the  epithelial  lining;  in  the  case  of 
ulceration,  the  necrotic  process  extends  beyond  the  basement 
membrane  to  involve  the  underlying  submucosa.  Adjacent  to 


Brown  et  al. — 447 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

the  area  of  necrosis,  there  is  usually  infiltration  of 
mononuclear  inflammatory  cells  and,  in  chronic  cases,  fibrous 
connective  tissue  proliferation.  Severity  grade  criteria: 

1  =  Small  focal  area  of  necrosis  less  than  the 

diameter  of  a  20X  microscopic  field. 

2  =  An  area  of  necrosis  less  than  the  diameter  of  a 

10X  microscopic  field. 

3  =  The  necrotic  area  extends  beyond  the  basement 

membrane  and  occupies  an  area  as  large  as  a  10X 
microscopic  field. 

4  =  The  ulcerated  area  is  greater  than  the  diameter 

of  a  4X  microscopic  field. 

SKELETAL  MPSCLE 

1.  ACUTE  MYOSITIS  -  This  is  acute  inflammation  of  skeletal 
muscle  tissue.  Muscle  fibers  are  separated  by  infiltration 
of  neutrophils  and  fewer  macrophages  and  lymphocytes.  In 
more  severe  cases  there  is  coagulative  necrosis  of  muscle 
fibers  and  phagocytosis  of  the  necrotic  cellular  debris. 
Severity  grade  criteria: 

1  *  Minimal  detectable  infiltration  of  neutrophils. 

2  *  Mild  infiltration  of  neutrophils  and  a  few 

macrophages  in  an  area  up  to  the  diameter  of  a 
10X  microscopic  field. 

3  *>  Coagulative  necrosis  of  muscle  tissue  with 

neutrophil  infiltration.  The  size  of  the 
necrotic  area  is  less  than  a  20X  microscopic 
field. 

4  =  Necrosis  of  muscle  and  neutrophilic 

infiltration.  The  necrotic  area  exceeds  the 
diameter  of  a  20X  field. 

2.  CHRONIC  MYOSITIS  -  Consists  of  chronic  inflammation  of 
skeletal  muscle.  Muscle  cells  are  separated  by  infiltration 
of  mononuclear  inflammatory  cells  in  which  large  macrophages 
predominate.  In  more  severe  cases  there  is  necrosis  of 
muscle  fibers  and  the  phagocytosis  of  necrotic  debris  by 
macrophages.  Muscle  fiber  regeneration  and  fibrous 
connective  tissue  proliferation  may  be  present.  Severity 
grade  criteria: 

1  =  Minimal  detectable  infiltration  of  mononuclear 
inflammatory  cells. 


Brown  et  al. — 448 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

70R  PATHOLOGY  FINDINGS 

2  Mild  infiltration  of  mononuclear  inflammatory 
cells.  Fibrosis  may  be  present.  The  lesion  is 
focal  and  does  not  exceed  the  diameter  of  a  10X 
microscopic  field. 

3  =  There  is  necrosis  of  muscle  cells  with 

phagocytosis  of  debris  by  macrophages. 
Regeneration  may  be  present.  The  area  of 
necrosis  is  less  than  that  of  a  20X  microscopic 
field. 

4  =  Lesion  is  similar  to  grade  3;  the  necrotic  area 

is  larger  than  a  20X  field. 

3.  CHRONIC  INFLAMMATION  RESTRICTED  TO  FASCIA  -  This  lesion 
consists  of  infiltration  of  lymphocytes,  macrophages,  and 
plasma  cells  into  the  fascia  dividing  bundles  of  skeletal 
muscle  fibers.  In  addition,  there  may  be  increased  amounts 
of  fibrous  tissue  in  these  septae.  There  is  no  damage  or 
inflammatory  cell  infiltration  into  adjacent  muscle  bundles. 
Severity  grade  criteria: 

1  =  Minimal  inflammatory  cell  infiltration 

involving  an  area  smaller  than  a  20X 
microscopic  field. 

2  «  Mild  inflammatory  cell  infiltration  involving 

an  area  smaller  than  a  10X  field. 

3  =  Inflammatory  cell  infiltration  involving  an 

area  larger  than  a  10X  field. 

4  =  Inflammatory  cell  infiltration  involving  an 

area  larger  than  a  4X  field. 

SCIATIC  NSRYtt 

No  lesions  were  observed  in  sciatic  nerves  of  rats  on 
this  study. 

TQHSUB 

1.  MONONUCLEAR  CELLULAR  INFILTRATION  -  There  is  focal, 
multifocal,  or  diffuse  infiltration  of  mononuclear 
inflammatory  cells  between  skeletal  muscle  fibers  in  the 
tongue.  Severity  grades  are  based  on  subjective  assessment 
of  the  number  of  cells  present: 

1  =  Minimal 

2  -  Mild 

3  =  Moderate 

4  Marked 


Brown  et  al. — 449 


Appendix  I  (cont . )  :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

2.  PERIARTERITIS  -  Inf lanunatory  cell  infiltration  of  the 
adventitia  of  small  to  medium-sized  arteries  and  adjacent 
tissues.  Severity  grades  are  based  on  subjective  assessment: 

1  =  Minimal 

2  =• Mild 

3  =  Moderate 

4  =  Marked 

3.  MEDIAL  HYPERTROPHY  AND  MINERALIZATION  OF  MEDIUM-SIZED 
MUSCULAR  ARTERIES  -  There  is  thickening  of  the  muscular  layer 
of  the  arterial  wall.  Several  vessels  in  the  same  tongue 
section  are  often  affected.  Hyali nization  and  mineralization 
of  hypertrophied  muscle  may  be  present.  The  lesion  is  coded 
as  "Present." 

SKIN 

1.  COMPOUND-RELATED  INFLAMMATORY  REACTION  -  See  "General 
Diagnoses"  above. 

2.  INFLAMMATORY  EXUDATE  ON  SKIN  SURFACE  -  There  is  a  mixture 
of  neutrophils,  macrophages,  fibrin,  and  cellular  debris  on 
the  surface  of  the  skin.  Severity  grades  are  based  on  the 
amount  of  surface  area  involved  relative  to  different 
microscopic  magnifications: 

1  =  Area  is  less  than  the  diameter  of  a  20X  field. 

2  =  Area  is  less  than  the  diameter  of  a  10X  field. 

3  =  Area  is  less  than  the  diameter  of  a  4X  field. 

4  -  Area  is  larger  than  the  diameter  of  a  4X  field. 

2.  PROCEDURE-RELATED  GRANULOMATOUS  CELLULITIS,  SUBCUTIS  - 
Compound- related  inflammatory  reactions  in  the  subcutis  are 
similar  to  those  in  other  locations.  See  "General  Diagnoses" 
above . 

3.  NONSUPPURATIVE  DERMATITIS  -  There  is  a  subacute 
inflammatory  process  involving  the  epidermis  and  dermis.  The 
predominant  change  involves  infiltration  of  lymphocytes, 
macrophages,  plasma  cells,  and  fewer  neutrophils  and 
eosinophils  into  these  areas.  The  inflammatory  cells  are 
present  within  the  epidermis,  along  the  basement  membrane, 
and  between  adnexal  structures  in  the  dermis .  Severity 
grades  are  based  on  the  amount  of  inflammatory  cell 
infiltration : 


Brown  et  al. — 450 


Appendix  I  (eont . ) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

1  -  Minimal  detectable  inflammatory  cell 

infiltrate . 

2  =  Mild  diffuse  infiltrate. 

3  =  Moderate  numbers  of  inflammatory  cells 

4  =  Affected  area  is  nearly  solidly  infiltrated. 

4 .  CELLULITIS  -  An  acute,  subacute,  or  chronic  inflammatory 
process  in  subcutaneous  tissue  characterized  by  inflammatory 
cell  infiltration.  Necrosis,  cellular  debris,  and  fibrin  may 
be  present  in  more  severe  cases.  No  te3t  material  or 
foreign-body  giant  cells  are  present.  Severity  grade 
criteria : 

1  *  Minimum  detectable  inflammatory  cell 

infiltration  in  the  subcutaneous  tissues. 

2  =  Mild  inflammatory  cell  infiltration.  May  be 

focal  (less  than  the  diameter  of  a  10X  field) 
or  diffuse. 

3  =  Moderate  numbers  of  inflammatory  cells 

distributed  either  focally  or  diffusely.  Small 
foci  of  necrosis,  less  than  the  diameter  of  a 
4 OX  field  may  be  present. 

4  *  Marked  inflammatory  cell  infiltration  and/or 

large  areas  of  necrosis. 

5.  KERATOACANTHOMA  (INVERTED  PAPILLOMA),  ANATOMIC  SITE  TO  BE 
SPECIFIED  -  A  benign,  well-encapsulated  tumor,  typically 
raised  above  the  surrounding  epidermis,  with  a  central  crater 
or  pore  that  is  filled  with  keratin.  The  neoplastic 
epidermis  lining  the  crater  contains  many  keratinized  pearls, 
which  are  particularly  numerous  near  the  center  of  the  tumor. 
There  is  sharp  demarcation  between  tumor  epithelium  and  the 
surrounding  dermis.  The  anatomic  location  is  documented 
using  the  special  histologic  comment  feature  of  the  Xybion 
program. 

6.  EROSION/ULCERATION  OF  THE  SKIN  SURFACE  -  An  erosion  is 
necrosis  and  sloughing  of  the  surface  epithelium.  The 
basement  membrane  is  intact.  An  ulceration  is  necrosis  of 
the  skin  which  extends  beyond  the  basement  membrane  to 
involve  the  underlying  dermis.  Usually,  there  is 
infiltration  of  neutrophils,  lymphocytes,  and  macrophages  at 
the  periphery  of  such  lesions.  In  chronic  cases  there  may  be 
fibrosis  and  attempts  at  epithelial  regeneration  as  well. 

The  lesion  is  coded  as  "Present." 


Brown  et  al. — 451 


Appendix  I  (eont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

7.  BASAL  CELL  TUMOR  -  Benign  tumor  of  basal  cell  origin. 

Basal  cells  are  usually  arranged  in  long  serpentine  ribbons 
or  small  nests.  The  tumors  are  well  demarcated  from  adjacent 
tissues  in  the  dermis.  The  tumor  cells  are  small  with  small 
amounts  of  lightly  basophilic  cytoplasm  and  round  to  oval 
vesicular  nuclei.  Mitotic  figures  are  rare.  There  is  a 
minimal  amount  of  fine  fibrous  stroma.  There  is  no  evidence 
of  invasion  or  metastasis. 

8.  BASAL  CELL  CARCINOMA  -  A  malignant  neoplasm  of  basal  cell 
origin.  Cellular  morphology  and  growth  patterns  are  very 
similar  to  the  benign  basal  cell  tumor.  Criterion  that 
distinguishes  malignant  tumors  is  their  invasiveness  of 
adjacent  tissue.  Tumor  cells  are  not  confined  by  the 
enveloping  capsule  and  invade  adjacent  tissue. 

9.  FIBROMA  -  Benign  tumor  of  fibrous  connective  tissue. 

These  are  well-demarcated  proliferations  of  well- 
differentiated  fibrocytes.  Cells  are  arranged  in  parallel 
fashion  or  interlacing  bundles.  Nuclei  are  small  and 
elongated.  Mitotic  figures  are  rare.  These  tumors  are  not 
highly  cellular  and  contain  an  abundance  of  mature  collagen. 

10.  ACANTHOSIS  -  A  non-neoplast ic  thickening  of  the 
epidermis.  The  entire  layer  of  the  epidermis  is  thickened. 
The  rete  ridges  are  elongated,  irregular,  and  extend  into  the 
dermis  for  a  variable  distance.  Severity  grade  criteria: 

1  =  Epidermis  is  twice  normal  thickness. 

2  =  Epidermis  is  three  times  normal  thickness. 

3  =  Epidermis  is  four  times  normal  thickness. 

4  =  Epidermis  is  more  than  four  times  normal 

thickness . 

11.  HYPERKERATOSIS  -  Thickening  of  the  stratum  corneum  by 
keratin  accumulation.  Severity  grade  criteria: 

1  -  Stratum  corneum  is  twice  normal  thickness. 

2  =  Stratum  corneum  is  three  times  normal 

thickness . 

3  *  Stratum  corneum  is  four  times  normal  thickness. 

4  «  Stratum  corneum  is  more  than  four  times  normal 

thickness . 


12.  SQUAMOUS  PAPILLOMA  -  A  small  superficial  pedunculated 
nodule  consisting  of  neoplastic  masses  of  mature  keratinizing 
epithelial  cells.  Cells  are  in  progressive  stages  of 


Brown  et  al . — 452 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

keratinization  often  occurring  in  concentric  whorls.  There 
is  no  evidence  of  deeper  invasion.  The  superficial  location 
and  the  mature,  highly  keratinized,  noninvasive  growth 
distinguish  papillomas  from  carcinomas.  These  tumors  have  a 
central  core  of  dermis  covered  by  layers  of  stratified 
squamous  epithelium  with  marked  hyperkeratosis  and 
parakeratosis . 

13.  PREPUTIAL  GLAND  ADENOMA  -  Well-differentiated,  benign 
tumor  of  the  preputial  gland.  Adenomas  are  composed 
primarily  of  glandular  tissue  with  small  nests  of  squamous 
cells  scattered  about.  Eosinophilic  granules  are  very 
prominent  in  the  cytoplasm  of  acinar  cells.  These  tumors 
grow  in  lobular  fashion  with  smooth  borders,  and  they  may 
become  very  large. 

14.  CLITORAL  GLAND  ADENOMA  -  Well-differentiated,  benign 
tumor  of  the  clitoral  gland,  the  female  counterpart  of  the 
male  preputial  gland  described  above.  Microscopic 
characteristics  are  similar  to  those  of  preputial  gland 
adenomas . 

15.  INFLAMMATION,  NOT  OTHERWISE  SPECIFIED,  PREPUTIAL  GLAND  - 
Presence  of  lymphocytes,  macrophages,  neutrophils,  or  plasma 
cells  within  the  preputial  gland.  This  lesion  is  coded  as 
"Present . " 

16.  NEUROFIBROSARCOMA,  SUBCUTANEOUS,  LOCATION  TO  BE  SPECIFIED 
-  Microscopically,  these  tumors  are  composed  of  spindle- 
shaped  cells  arranged  in  circular  whorls  or  wavy  fasciculi. 
They  closely  resemble  fibrosarcomas  but  differ  in  that  cells 
tend  to  have  the  form  of  plump  spindles  and  retain  the 
whorled,  curly  arrangement.  The  location  of  these  tumors  is 
documented  in  the  special  histologic  comments  feature  of  the 
Xybion  program. 

17.  FIBROSARCOMA  -  Malignant  neoplasm  of  fibrous  connective 
tissue.  Tumor  cells  with  their  collagenous  fibrils  are 
arranged  haphazardly  and  run  aimlessly  in  a  variety  of 
directions.  Fasciculi  of  cells  lying  in  the  same  plane  are 
often  seen.  A  high  degree  of  cellularity,  hyperchromat ic 
staining  of  nuclei,  and  a  tendency  for  the  nuclei  to  be 
plump,  stellate,  and  bizarre  indicate  malignancy.  Mitotic 
figures  may  be  very  numerous. 

18.  EPIDERMAL  INCLUSION  CYST  -  One  or  more  small  cysts 


Brown  et  al. — 453 


Appendix  Z  (cont.):  GLOSSARY  OF  TERMS 

TOR  PATHOLOGY  FINDINGS 

located  within  the  dermis  and  surrounded  by  a  thin  fibrous 
capsule.  The  cyst  wall  consists  of  flattened  squamous 
epithelium  without  adnexal  structures.  The  cyst  is  filled 
with  concentrically  laminated  masses  of  keratin  and  amorphous 
cellular  debris.  Intense  inflammatory  reaction  may  occur 
around  these  lesions  if  the  cyst  ruptures.  This  lesion  is 
coded  as  "Present . " 

MAMMARY  GLANDS 

1.  HYPERPLASIA  AND/OR  HYPERTROPHY  OF  ACINAR  TISSUE  - 
Increased  size  and  number  of  mammary  secretory  cells.  These 
cells  are  arranged  in  lobules.  In  hyperplastic  glands,  the 
lobules  become  larger  and  closer  together.  Cells  are 
cuboidal  to  low  columnar,  and  crowded  together  on  the 
basement  membrane  to  the  point  that  mild  piling  up  of  cells 
occurs.  Often  these  cells  are  actively  secreting  and  acini 
become  distended  with  slightly  basophilic  granular  or  foamy 
secretory  product .  Mammary  hyperplasia  occurs  in  both  sexes 
although  it  is  more  prevalent  and  usually  more  severe  in 
females.  The  same  severity  grade  criteria  are  used  for  both 
sexes : 

1  =*  Foci  of  acinar  cells  are  widely  scattered. 

Most  are  less  than  the  diameter  of  a  4 OX 
microscopic  field. 

2  -  Acinar  foci  are  more  numerous  but  small.  The 

largest  are  larger  tha:.  a  40X  field  but  smaller 
than  a  20X  field. 

3  *  Extensive  acinar  tissue  is  present.  It  is 

nearly  confluent  but  there  are  a  few  uninvolved 
areas . 

4  -  Confluent  secretory  tissue  is  present. 

2.  GALACTOCOELE  -  Marked  cystic  dilatation  of  a  mammary  duct. 
The  dilated  duct  is  lined  by  flattened  ductal  epithelium  and 
is  filled  with  secretory  product.  The  lesion  is  coded  as 
"Present . " 

3 .  FIBROADENOMA  -  A  benign  tumor  of  mammary  origin  in  which 
there  is  proliferation  of  both  epithelial  and  connective 
tissue  components.  Histologically,  fibroadenomas  vary 
considerably  in  their  structural  composition.  Either  the 
epithelial  or  the  connective  tissue  component  may 
predominate.  Usually  there  is  a  concomitant  growth  of 
connective  tissue  stroma,  glandular  and  ductal  cells.  Ducts 


Brown  et  al . — 454 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

may  become  surrounded  by  layers  of  fibrous  connective  tissue 
and  the  histological  appearance  becomes  that  of  single  acini 
or  tubules  separated  from  each  other  by  concentric  layers  of 
loose  connective  tissue.  The  secretory  epithelium  is  usually 
only  one  cell  layer  thick  and  bears  the  same  relation  to 
myoepithelium  and  basement  membrane  as  it  does  in  the  normal 
mammary  gland. 

4.  DILATATION  (ECTASIA)  OF  DUCTS  AND  DUCTULES  -  Dilatation 
of  mammary  ducts  occurs  independently  of  mammary  acinar 
hyperplasia  and  is  therefore  coded  independently.  Dilated 
ducts  often  contain  pale  eosinophilic  secretory  material. 

This  lesion  is  coded  as  "Present.” 

5.  MAMMARY  ADENOCARCINOMA  -  A  malignant  tumor  that  is  derived 
from  glandular  epithelium.  The  epithelium  is  arranged  in 
glandlike  structures  surrounding  a  lumen.  Acinar  spaces  may 
be  orderly  and  regular  or  may  vary  considerably  in  size  and 
shape.  Eosinophilic  or  basophilic  secretion  is  often  present 
in  glandular  spaces.  The  epithelium  varies  from  low  cuboidal 
to  tall  columnar  cells  and  may  be  several  cell  layers  thick. 
Nuclei  are  large  and  mitotic  figures  are  numerous.  In  some 
adenocarcinomas,  the  acinar  spaces  are  completely  filled  with 
tumor  cells  and  no  lumens  are  present.  Stroma  within  cell 
masses  is  usually  scanty.  Papillary  projections  can  form, 
however.  Stromal  invasion  by  neoplastic  epithelial  cells  is 
common.  Necrosis  and  mononuclear  inflammatory  cell 
infiltration  can  occur  and  are  more  frequent  in  larger 
tumors . 

HQSB/TVBBlHftlBa 

1.  ACUTE  INFLAMMATION  -  Presence  of  suppurative  inflammation 
within  the  nasal  cavity.  Necrosis  may  be  present  in  more 
severe  lesions.  Severity  grade  criteria: 

]  *  Minimal  detectable  infiltration  of  neutrophils. 
1  -  Mild  infiltration  of  neutrophils. 

3  *  Moderate  infiltration  of  neutrophils. 

Suppurative  exudate  is  present  within  nasal 
passages . 

4  *  Marked  neutrophilic  infiltration.  Suppurative 

exudate  is  present  in  nasal  passages.  Necrosis 
with  destruction  of  nasal  tissues  is  present. 


Brown  et  al. — 455 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

2.  SUBACUTE  INFLAMMATION  -  Infiltration  of  lymphocytes, 
macrophages,  and  plasma  cells  into  tissue  of  the  nasal 
cavity.  Necrosis  may  be  present  in  more  severe  lesions. 
Severity  grade  criteria: 

1  =  Minimal  infiltration  of  mononuclear 

inflammatory  cells  . 

2  =  Mild  mononuclear  cell  infiltration. 

3  =  Moderate  mononuclear  infiltration.  Exudate  is 

present  in  nasal  passages. 

4  =  Marked  mononuclear  cell  infiltration.  Exudate 

is  present  in  nasal  passages.  Necrosis  with 
destruction  of  nasal  tissues  is  present. 

3.  MIXED  INFLAMMATORY  CELL  INFILTRATION,  NASAL  CAVITY, 
SINUSES,  TURBINATES  OR  NASOLACRIMAL  DUCTS  -  This  diagnosis  is 
used  primarily  for  small  focal  inflammatory  lesions  in  the 
nasal  cavity.  The  exact  location  of  the  lesion  is  documented 
in  the  special  histologic  comments  of  the  Xybion  program. 

Most  of  the  lesions  coded  under  this  diagnosis  are 
inflammatory  foci  adjacent  to  nasolacrimal  ducts. 

Inflammatory  cell  foci  are  very  common  in  this  location  in 
rats  used  in  Study  GLP  83009.  Severity  grade  criteria: 

1  =  Minimal  inflammatory  cell  infiltration. 

2  =  Mild  inflammatory  cell  infiltration. 

3  =*  Moderate  inflammatory  cell  infiltration; 

cellular  exudate  is  present  within  a  natural 
anatomic  space,  e.g.,  the  lumen  of  the 
nasolacrimal  duct. 

4  =  Marked  inflammatory  cell  infiltrate.  Exudate 

is  present  within  a  natural  anatomic  space. 
Small  amounts  of  necrosis  may  be  present. 

BONE 

Bone  and  bone  marrow  were  examined  from  the  STERNUM. 
FEMUR,  and  VERTEBRAE  as  well  as  incidental  locations  such 
as  th«  nasal  cavity,-  and  middle  and  inner  ear.  The  diagnoses 
coded  for  bone  marrow  from  all  these  sites  are  described 
below. 

1.  MYELOID  HISTIOCYTOSIS  -  This  is  an  unusual  lesion  that 
affects  bone  marrow  uniformly  in  an  affected  animal.  The 
marrow  contains  large  numbers  of  large  histiocyte-like  cells. 
No  microorganisms  were  detected  in  sections  stained  by 


Brown  et  al . — 456 


Appendix  Z  (cont. ) :  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

Gomori's  Mathenamine  Silver  stain.  Periodic  Acid-Schiff 
(PAS),  Giemsa,  or  Goodpasture  stains.  Cells  contained 
diastase-resistant  PAS-positive  material.  Whether  this 
lesion  is  inflammatory  or  neoplastic  in  nature  is  unknown. 

The  lesion  is  coded  as  "Present.” 

2.  HYPERPLASIA,  MYELOID  -  This  change  in  bone  marrow  consists 
of  excess  non-neoplastic  proliferation  of  granulocytic 
precursors.  Erythroid  precursors  are  present  but  usually 
reduced  in  number.  The  myeloid  proliferation  may  be 
multifocal  or  diffuse.  The  change  is  coded  as  "Present." 

3.  MYELOFIBROSIS/OSTEOSCLEROSIS  -  All  normal  marrow  elements 
are  greatly  reduced  in  number  and  are  replaced  by 
hypocellular  loosely  arranged  fibrous  tissue.  The  lesion  is 
coded  as  "Present." 

4 .  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  See  "General 
Diagnoses"  above. 

SPINAL  CORD 

No  gross  or  microscopic  lesions  were  observed  the  the 
spinal  cords  of  rats  used  in  Study  GLP  83009. 

ABRSHAL  GLANDS 

1.  CORTICAL  ADENOMA  -  A  benign  tumor  of  adrenal  cortical 
cells.  The  tumor  cells  are  arranged  in  a  discrete  nodule 
which  compresses  adjacent  cortical  tissue.  Within  the  tumor, 
the  cells  lack  their  normal  orderly  arrangement  and 
relationship  to  basement  membrane  and  vasculature.  They  are 
arranged  in  short  cords,  or  nests  separated  by  delicate 

f ibrovascular  stroma.  Tumor  cells  may  be  either  larger  or 
smaller  than  their  non-neoplastic  counterparts  and  the 
tinctorial  qualities  of  their  cytoplasm  vary  considerably. 

The  presence  of  lipid  vacuoles  is  quite  common  as  is  slight 
granular  basophilia.  Mitotic  figures  are  rare.  Adenomas  are 
well-vascularized  and  irregular,  blood-filled,  endothelial- 
lined  spaces  and  hemorrhage  are  common  findings. 

2.  LIPIDOSIS  -  Presence  of  intracytoplasmic  lipid  vacuoles  in 
cortical  cells.  This  change  may  be  uniform  or  multifocal. 
Severity  grade  is  based  on  the  amount  of  the  adrenal  cortex 
involved: 


Brown  et  al. — 457 


Appendix  I  (cont.) :  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

1  *  Less  than  25%  of  the  cortical  cells  are 

vacuolated. 

2  *  25%  to  50%  of  the  cells  are  affected. 

3  =  50%  to  75%  of  the  cortex  is  affected. 

4  =  More  than  75%  of  the  cortex  is  affected. 

3.  FOCI  OF  CELLULAR  ALTERATION  -  These  are  degenerative 
lesions  affecting  cells  of  the  adrenal  cortex.  The  changes 
are  focal.  Affected  cells  may  be  either  larger  or  smaller 
than  normal  and  have  different  tinctorial  qualities  as 
follows : 

3A.  FOCUS  OF  CELLULAR  ALTERATION,  EOSINOPHILIC  -  Degenerative 
cortical  cells  are  slightly  larger  than  normal  and  their 
cytoplasm  is  granular  and  eosinophilic.  There  is  slight 
compression  of  adjacent  normal  cortical  cells. 

3B.  FOCUS  OF  CELLULAR  ALTERATION,  VACUOLATED  -  Degenerative 
cortical  cells  may  be  slightly  larger  than  normal  and  their 
cytoplasm  is  vacuolated.  There  is  usually  slight  compression 
of  adjacent  cortical  cells. 

3C .  FOCUS  OF  CELLULAR  ALTERATION,  BASOPHILIC  -  Degenerative 
cortical  cells  may  be  slightly  smaller  than  normal  and  their 
cytoplasm  has  a  slight  basophilic  granularity.  Severity 
grade  criteria  for  foci  of  cellular  alteration: 

1  =  One  focus . 

2  *  Two  or  three  foci . 

3  *  Four  to  six  foci. 

4  *  More  than  six  foci. 

4.  CORTICAL  NODULAR  HYPERPLASIA  -  Nodular  proliferation  of 
adrenal  cortical  cells.  Cells  may  be  slightly  larger  than 
normal  and  cytoplasm  may  be  finely  vacuolated.  These  small 
nodules  are  usually  located  at  the  surface  of  the  adrenal  and 
are  surrounded  by  a  thin  extension  of  the  adrenal  capsule. 
This  change  is  coded  as  "Present." 

5.  FIBROSIS,  CAPSULE  -  Proliferation  of  fibrous  connective 
tissue  in  the  adrenal  capsule.  There  are  minimal  numbers  of 
mononuclear  inflammatory  cells  present.  Severity  grade 
criteria  are  based  on  the  proportion  of  the  adrenal  capsule 
involved: 


1  -  Minimal  capsular  fibrosis;  less  than  10%  of 
capsule  is  involved. 


Brown  et  al . — 458 


Appendix  I  (cont.) :  GLOSSARY  Or  TERMS 

FOR  PATHOLOGY  FINDINGS 

2  **  10%  to  25%  cf  the  capsule  is  involved. 

3  *  25%  to  50%  of  the  capsule  is  fibrotic. 

4  =  More  than  50%  of  the  capsule  is  fibrotic. 

6.  MEDULLARY  CARCINOMA  (MALIGNANT  PHEOCHROMOCYTOMA)  - 
Malignant  tumor  of  the  adrenal  medulla.  The  cells  resemble 
normal  medullary  cells  but  usually  they  have  less  cytoplasm 
and  the  nuclei  are  hyperchromatic .  The  cells  are  arranged  in 
nests  and  sometimes  in  trabecular  fashion.  Usually  the  cells 
are  of  one  type  although  there  are  occasional  small 
basophilic  cells,  with  transitions  to  larger  cells  having 
abundant  cytoplasm  and  large  nuclei.  The  only  criteria  that 
are  used  to  differentiate  malignant  from  benign  tumors  of  the 
adrenal  medulla  is  invasion  of  the  capsule  and  infiltration 
of  tissues  outside  the  adrenal  gland  or  metastasis. 

7.  PHEOCHROMOCYTOMA  -  Benign  tumor  of  the  adrenal  medulla. 
Morphologic  characteristics  are  similar  to  that  of  medullary 
carcinoma  described  above.  The  benign  tumors  may  become 
quite  large  but  are  confined  to  the  adrenal  gland. 

8 .  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  See  "General 
Diagnoses"  above. 

9.  EXTRAMEDULLARY  HEMATOPOIESIS  -  Presence  of  erythrocytic 
precursors  within  the  adrenal  gland.  This  change  is  coded  as 
"Present . " 

10.  NECROSIS  -  Coagulative  or  liquefactive  necrosis  in  the 
adrenal  cortex  or  medulla.  Severity  grade  criteria: 

1  ■  Focus  of  necrosis  less  than  the  diameter  of  a 

40X  microscopic  field. 

2  =  Necrosis  of  less  than  25%  of  an  adrenal  gland. 

3  =  Necrosis  of  25%  to  50%  of  an  adrenal  gland. 

4  =  Necrosis  of  more  than  50%  of  an  adrenal  gland. 

PITUITARY  GLAND 

1.  ADENOMA  -  A  benign  tumor  of  the  anterior  pituitary  gland. 
Proliferating  cells  of  the  anterior  pituitary  gland  vary 
considerably  in  their  cytological  characteristics  and  in 
their  tinctorial  qualities.  Using  only  hematoxylin  and  eosin 
staining,  it  is  not  possible  to  identify  cell  types 
accurately  or  to  identify  the  type  of  hormone  that  may  be 


Brown  et  al. — 459 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

TOR  PATHOLOGY  FINDINGS 

secreted.  Pituitary  tumor  cells  are  usually  arranged  in 
short  cords  or  nests  separated  by  very  fine  f ibrovascular 
stroma.  They  are  highly  vascular  and  irregular,  blood- 
filled,  endothelial-lined  spaces  occur  frequently  as  does 
hemorrhage.  Metastases  occur  rarely  but  large  tumors  may 
infiltrate  adjacent  tissue  and  compress  overlying  tissue  of 
the  brain  which  undergoes  compression  atrophy. 

2.  CYST(S)  -  Presence  of  spaces  within  the  pituitary  that  are 
usually  filled  with  pale  eosinophilic  fluid.  The  spaces  may 
be  lined  with  endothelium  or  flattened  epithelial  cells. 
Rarely,  a  ciliated  cell  may  be  observed.  Cysts  in  the 
pituitary  may  be  single  or  multiple.  They  are  coded  as 
"Present . " 

3.  ADENOCARCINOMA  -  A  malignant  tumor  of  the  anterior 
pituitary  gland.  Morphologically,  malignant  tumors  of  the 
pituitary  can  be  distinguished  from  their  benign  counterparts 
only  by  their  invasiveness.  Metastases  are  very  rare.  If 
the  tumor  cells  infiltrate  tissues  outside  the  confines  of 
the  pituitary  gland,  it  is  coded  as  an  adenocarcinoma. 

4.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  See  "General 
Diagnoses"  above. 

EYES  AND  OPTIC  NERVE 

1.  CORNEAL  MINERALIZATION  -  Presence  of  small  spicules  of 
mineral  within  the  cornea.  Usually  this  is  accompanied  by 
vascularization  and  occasionally  by  infiltration  of  a  few 
inflammatory  cells.  Severity  grade  criteria: 

1  *  Presence  of  one  focus  of  mineral. 

2  =  Presence  of  two  or  three  mineralized  foci. 

3  =  Four  to  six  foci  present. 

4  =  More  than  six  mineralized  foci  present. 

2.  RETINAL  ATROPHY  AND/OR  DEGENERATION  -  Consists  of  variable 
reduction  of  cellular  elements  and  disorganization  of  the 
different  layers  of  the  retina.  This  is  a  progressive 
condition  that  increases  in  severity  as  the  animal  ages .  The 
condition  may  be  unilateral  or  bilateral.  Severity  grade 
criteria: 


1  ~  Perceptible  reduction  in  the  number  of  cells  in 
the  retina. 


Brown  et  al . — 460 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

2  *  Multifocal  disorganization  of  retinal  layers. 

3  =  Diffuse  disorganization  of  retinal  layers. 

Some  parts  of  layers  may  be  recognizable. 

4  =  No  recognizable  retinal  tissue  is  present.  The 

retina  consists  of  a  band  of  fibrous  tissue. 

3.  CORNEAL  PIGMENTATION  -  Presence  of  pigment  within  the 
cornea.  Pigment  may  be  hemosiderin  or  melanin.  It  is 
present  within  histiocytic  cells  and  usually  accompanies 
inflammatory  lesions  in  the  cornea.  Such  pigment 
accumulations  are  usually  focal.  Severity  grade  criteria  is 
based  on  the  number  of  pigmented  foci  present: 

1  =  One  pigmented  focus  is  present. 

2  =  Two  or  three  pigmented  foci  are  present . 

3  =  Four  to  six  pigmented  foci  are  present. 

4  =  More  than  six  pigmented  foci  are  present. 

4 .  CORNEAL  VASCULARIZATION  -  Presence  of  endothelial-lined 
vascular  spaces  beneath  the  epithelium  or  within  the  corneal 
stroma.  Severity  grade  criteria  are  based  on  the  number  of 
vascular  cross  sections  present: 

1  =  One  vascular  cross  section. 

2  *  Two  or  three  vascular  cross  sections. 

3  =  Four  to  six  vascular  cross  sections. 

4  =  More  than  six  vascular  cross  sections. 

5 .  CHRONIC  KERATITIS  -  Chronic  inflammation  involving  the 
cornea.  One  or  more  of  the  following  histologic  findings  may 
be  present:  mononuclear  inflammatory  cell  infiltration 
beneath  the  epithelium  and  into  the  stroma,  erosion  or 
ulceration  of  the  epithelium,  proliferation  of  vascular 
structures  (see  corneal  vascularization  above) ,  presence  of 
pigment,  necrosis  of  cellular  elements,  and  presence  of  scar 
tissue.  Severity  grade  criteria  are  based  on  the  degree  of 
corneal  damage  and  the  amount  of  the  cornea  involved: 

1  -  Minimal  to  mild  mononuclear  inflammatory  cell 

infiltration . 

2  -  Moderate  inflammatory  cell  infiltrate  with 

vascular  proliferation  or  pigment  deposition. 

3  =  Similar  to  grade  2  plus  minimal  necrosis  or 

scar  tissue  proliferation. 

4  =  Marked  chronic  inflammatory  changes  involving 

50%  or  more  of  the  cornea. 


Brown  et  al. — 461 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

70R  PATHOLOGY  FINDINGS 

6.  IRIDOCYCLITIS  -  Inf lammation  of  the  iris  and/or  ciliary 
body.  This  was  a  very  unusual  lesion  in  this  study.  It 
affected  only  three  rats  in  which  it  consisted  of  mild 
infiltration  of  the  uveal  tract  with  mononuclear  inflammatory 
cells.  Severity  grade  criteria: 

1  =  Minimal  infiltration  of  uveal  tract  with 

mononuclear  inflammatory  cells. 

2  =  Mild  mononuclear  infiltration  of  uveal  tract. 

3  =  Criteria  not  established. 

4  =  Criteria  not  established. 

7.  METAPLASTIC  BONE  FORMATION  AND/OR  MINERALIZATION,  SCLERA  - 
This  very  common  finding  consists  of  focal  proliferation  of 
plaques  of  metaplastic  bone  or  deposition  of  mineral,  or 
both,  in  the  dense  fibrous  tissue  of  the  sclera.  There  are 
no  inflammatory  lesions  associated  with  these  changes.  These 
changes  are  coded  as  "Present." 

8.  LENTICULAR  DEGENERATIVE  CHANGES  COMPATIBLE  WITH  CATARACT  - 
A  cataract  is  opacity  of  the  lens  that  can  be  detected 
grossly.  Microscopically,  this  lesion  may  consist  of  some  or 
all  of  the  following  degenerative  changes  in  the  lens: 
abnormal  proliferation  of  lens  epithelium  which  may  become 
redundant  and  disorganized,  disorganization  of  lens  fibers  in 
the  cortex  with  aggregation  into  spherical  masses  or  globules 
of  material  which  may  become  mineralized,  increased  density, 
disorganization,  and  possibly  liquefaction  of  nuclear  fibers. 
These  changes  are  coded  collectively  under  this  diagnosis  and 
are  designated  as  "Present.” 

9.  CHRONIC  CONJUNCTIVITIS  -  Chronic  inflammation  of  the 
conjunctiva.  This  lesion  is  characterized  primarily  by  the 
infiltration  of  mononuclear  inflammatory  cells  into  the 
conjunctival  membrane.  Fibrosis  may  be  present.  Severity 
grade  criteria  are  based  on  the  amount  of  cellular 
infiltration  present: 

1  *  Minimal  mononuclear  cell  infiltrate. 

2  -  Mild  mononuclear  cell  infiltrate. 

3  -  Moderate  mononuclear  cell  infiltrate. 

4  =  Marked  mononuclear  cell  infiltrate. 

EAR 

1.  OTITIS  MEDIA,  SUPPURATIVE  OR  NONSUPPURATIVE  -  Acute  or 


Brown  et  al. — 462 


Appendix  I  (cont.):  GLOSSARY  OF  TERMS 

FOR  PATHOLOGY  FINDINGS 

subacute  inflammation  of  middle  ear  structures. 
Microscopically,  this  lesion  consists  of  infiltration  of 
neutrophils  (suppurative)  or  mononuclear  inflammatory  cells 
(nonsuppurative)  into  the  structures  of  the  middle  ear. 
Severity  grade  criteria: 

1  =  Minimal  inflammatory  cell  infiltration. 

2  =  Mild  inflammatory  cell  infiltration  into 

tissues  surrounding  the  cavities  of  the  middle 
ear . 

3  =  Moderate  inflammatory  cell  infiltrate;  presence 

of  cellular  exudate  within  the  cavities  of  the 
middle  ear. 

4  =  As  in  Grade  3  above  plus  necrosis  and 

destruction  of  middle  ear  structures. 

2.  FIBROSARCOMA,  SUBCUTANEOUS,  EXTERNAL  EAR,  ADJACENT  TO  EAR 
TAG  -  Histological  characteristics  of  this  tumor  are  similar 
to  fibrosarcomas  described  under  the  SKIN.  There  was  one 
such  tumor  adjacent  to  the  ear  tag  in  one  animal. 

3.  PITUITARY  TUMOR  INVASION  -  There  is  an  infiltration  of 
cells  from  a  pituitary  tumor  within  the  cranial  cavity 
adjacent  to  the  structures  of  the  middle  and  inner  ear  near 
the  origin  of  the  acoustic  nerve.  Cellular  characteristics 
and  arrangement  of  tumor  cells  are  similar  to  that  described 
for  adenoma  under  the  PITUITARY  GLAND. 

APPITQRX  SEBACEOUS  GLAND 

No  gross  or  microscopic  lesions  were  recognized  in  the 
auditory  sebaceous  glands  of  rats  in  Study  GLP  83009. 

ABPQMINAL  WALL 

1.  INFECTION  AND  INFLAMMATION,  SEVERE,  INVOLVING  MUSCULATURE 
AND  PERITONEAL  SURFACE  -  Thl.3  diagnosis  is  self-explanatory. 
This  lesion  was  not  observed  in  animals  necropsied  during  the 
second  year  of  this  study.  This  diagnosis  is  described  in 
the  interim  report  of  Study  GLP  83009,  which  covers  the 
findings  of  the  first  year  of  this  two-year  study. 

THORAX 

1.  PRESENCE  OF  METASTATIC  SARCOMA  IN  THORACIC  WALL  -  See 
"General  Diagnoses"  above. 


Brown  et  al . — 463 


Appendix  I  (cont.):  GLOSSARY  07  TERMS 

FOR  PATHOLOGY  FINDINGS 

HBSI1  BODX 

1.  LARGE  GRANULAR  LYMPHOCYTE  LEUKEMIA  -  Since  neoplastic 
cells  from  this  disease  may  occur  in  virtually  every  organ 
and  tissue  of  the  body,  the  primary  site  for  the  lesion  is 
coded  under  WHOLE  BODY.  See  "General  Diagnoses"  above  for 
microscopic  description  and  criteria  for  coding  metastases. 

2.  INFLAMMATION  OF  BROWN  FAT  -  This  lesion  consists  of 
infiltration  of  inflammatory  cells  and  proliferation  of 
fibrous  connective  tissue  in  brown  fat.  The  lesion  can  occur 
in  brown  fat  deposits  anywhere  in  the  body.  For  this  reason 
it  is  coded  under  WHOLE  BODY  and  its  location  is  specified  in 
the  special  histologic  comments  section  of  the  Xybion 
program.  The  lesion  is  coded  as  "Present." 

MEDIASTINUM 

1.  CHRONIC  INFLAMMATION  -  The  presence  of  lymphocytes, 
macrophages,  plasma  cells,  and  the  proliferation  of  fibrous 
connective  tissue  within  the  mediastinal  tissues.  Severity 
grade  is  based  on  a  subjective  impression  of  the  intensity 
and  extent  of  the  inflammatory  lesion: 

1  -  Minimal  detectable  infiltration  of  inflammatory 

cells . 

2  =  Mild  inflammatory  response  limited  to  a  small 

focal  area  in  the  mediastinum. 

3  -  Moderate  inflammatory  response;  not  all  tissues 

in  the  mediastinum  are  involved. 

4  =  Marked  inflammatory  response  involving  most  of 

the  tissues  in  the  mediastinum. 

2.  REACTIVE  HYPERPLASIA,  LYMPH  NODE  -  Lymph  nodes  in