Appendix_5__Sample_QA_Report.pdf

PATHOLOGY QUALITY ASSESSMENT REPORT

FOR

CHRONIC GAVAGE TOXICITY/CARCINOGENICITY

STUDY OF

IN MALE AND FEMALE B6C3F1 MICE

CHEMICAL/TDMS NO.:

VOLUME I OF III

Conducted by:

Submitted to:

National Toxicology Program Research Triangle Park, NC 27709

Submitted by:

Research Triangle Park, NC 27709

Under Contract No.:

TABLE OF CONTENTS

VOLUME I

Page

Signature Page ...................................................................................................... i

Photography Schedule ............................................................. ii

Quality Assessment Narrative

Conflict of Interest Statement

APPENDIX A

Observations and Organ Notes to be Addressed During Pathology Quality A-1

Assessment

APPENDIX B

Slide Inventory Records ................................................................................. B-1

VOLUME II

APPENDIX C

Slide Review Work Sheets - Males ................................................................ C-1

VOLUME III

APPENDIX D

Slide Review Work Sheets - Females ............................................................ D-1

DISCLAIMER(S)

This document was produced to meet the requirements of Section 508 of the Rehabilitation

Act and if you are unable to view or interpret these, please contact 919-998-9407 or

. Documents produced by third parties are provided for your convenience; hence, some portions of this report may not meet the requirements of

Section 508.

SIGNATURE PAGE

Signature Page for

Chemical Name

C# TDMS# Test# Species B6C3F1 MICE

Lab Study CHRONIC Route GAVAGE

Entries in the "PWG Coordinator's Comments/PWG Consensus" (PWG) column and in the "Action To Be Taken" (AT) column of the QA worksheets were made by the PWG Coordinator. Entries in the PWG column represent the diagnosis of the coordinator and when ** is present, the diagnosis of the Pathology Working Group as well. Entries in the AT column are directives for changes in the pathology data which reflect the consensus diagnoses resulting from the peer review process. No entry in the PWG column indicates the lesion/tissue was not reviewed by the PWG Coordinator or PWG; no entry in the AT column indicates that no changes are necessary.

PWG Coordinator date

Each entry in the "Action To Be Taken" column has been reviewed and approved by the NTP/NIEHS pathologist assigned to this chemical/agent or their designee. The NTP/NIEHS pathologist or designee has made a red check in the extreme right border for each entry, and has initialed and dated (in red) the bottom of each page on which at least one entry was made.

NTP/NIEHS Pathologist / Designee date

As directed by the entries in the "Action To Be Taken" column, the pathology data stored in the Toxicology Data Management System (TDMS) has been updated by the data entry technician via a TDMS administrative work station.

Data Entry Technician date

As directed by the Post PWG Audit comments, the pathology data stored in the Toxicology Data Management System (TDMS) has been updated by the data entry technician via a TDMS administrative work station.

Data Entry Technician date i

PHOTOGRAPHY SCHEDULE

PHOTO. # CHEM. # SPECIES GROUP ANI. # HISTO. # TISSUE LESION MAG.

M3 HM 155 NOSE, L3 RESPIRATORY METAPLASIA 4

M3 HM 155 NOSE, L3 RESPIRATORY METAPLASIA 16

M3 HM 155 NOSE, L3 RESPIRATORY METAPLASIA 32

M3 HM 155 NOSE, L3

RESP EPITH, HYALINE DROPLET ACCUMULATION,

NASOPHARYNGEAL DUCT 32

M3 HM 155 NOSE, L3 OLF EPITH, HYALINE DROPLET ACCUMULATION 32

M3 HM 155 NOSE, L3 OLF EPITH, HYALINE DROPLET ACCUMULATION 32

M3 HM 155 NOSE, L3 OLF EPITH, ATROPHY 32

M3 HM 155 NOSE, L3

OLF EPITH, SQUAMOUS METAPLASIA; OLF NERVE,

ATROPHY 32

M3 HM 155 NOSE, L3

OLF EPITH, RESPIRATORY METAPLASIA; OLF NERVE,

ATROPHY 32

M3 HM 155 NOSE, L3 OLF EPITH, NECROSIS 32

M3 HM 155 NOSE, L3 OLF EPITH, ATROPHY 32

M3 HM 155 NOSE, L3 RESP EPITH, HYPERPLASIA & INFLAMMATION 32

M3 HF 361

STOMACH,

GLANDULAR HYPERPLASIA; INFLAMMATION, CHRONIC; PIGMENT 6.4

M3 HF 361

STOMACH,

GLANDULAR HYPERPLASIA; INFLAMMATION, CHRONIC; PIGMENT 16

M3 HF 361

STOMACH,

GLANDULAR HYPERPLASIA; INFLAMMATION, CHRONIC; PIGMENT 32

M3 VM 10

STOMACH,

GLANDULAR NORMAL 6.4

M3 VM 10

STOMACH,

GLANDULAR NORMAL 16

ii

PHOTO. # CHEM. # SPECIES GROUP ANI. # HISTO. # TISSUE LESION MAG.

M3 VM 10

STOMACH,

GLANDULAR NORMAL 32

M3 VM 10 NOSE, L3 NORMAL 4

M3 VM 10 NOSE, L3 OLF EPITH, NORMAL; OLF NERVE, NORMAL 16

M3 VM 10 NOSE, L3 OLF EPITH, NORMAL; OLF NERVE, NORMAL 32

M3 VM 10 NOSE, L3 OLF EPITH, NORMAL 32

M3 VM 10 NOSE, L3 RESP EPITH, NORMAL, NASOPHARYNGEAL DUCT 32

M3 VM 10 NOSE, L2 OLF EPITH, NORMAL 32

M3 VM 14 BRAIN GRANULAR CELL TUMOR 2

M3 VM 14 BRAIN GRANULAR CELL TUMOR 10

M3 VM 14 BRAIN GRANULAR CELL TUMOR 20

M3 VM 14 BRAIN GRANULAR CELL TUMOR 32

M3 LF 253 BRAIN MENINGIOMA 2

M3 LF 253 BRAIN MENINGIOMA 10

M3 LF 253 BRAIN MENINGIOMA 16

M3 LF 253 BRAIN MENINGIOMA 16

M3 LF 253 BRAIN MENINGIOMA 32

M3 LF 253 BRAIN MENINGIOMA 32

M3 VM 10 NOSE, L3 OLF EPITH, NORMAL 32

iii

NARRATIVE

PATHOLOGY QUALITY ASSESSMENT NARRATIVE

FOR

CHRONIC GAVAGE TOXICITY/CARCINOGENICITY

STUDY OF

CHEMICAL NO./TDMS NO.:

INTRODUCTION

The original Post Experimental Information System (PEIS) Individual Animal

Necropsy Records (IANRs), PEIRPT02, 03, 04, 08, 09, 10, 11, 14, 17 and 18, EISRPT 01, 02, 04, 05 and 12, the Toxicology Report, and hematoxylin and eosin stained microscope slides used in the Chronic Gavage Toxicity and Carcinogenicity Study of in Male and Female B6C3F1 Mice were received by

. The tissues had previously been examined and the data reported by a pathologist at . The data and slides were reviewed by an and the results are summarized in this report.

The experimental design for this study is as follows:

Experimental Design

CHRONIC TOXICITY STUDY

DOSAGE

(MG/KG)

B6C3F1 MICE

MALE FEMALE

0 50 50

62.5 50 50 125 50 50 250 50 50

Final

BACKGROUND INFORMATION/ SUMMARY OF ORIGINAL STUDY RESULTS

The following information is excerpted from the Chronic Gavage Toxicity/

Carcinogenicity Study of Mice dated and submitted to the National Institute of Environmental Health Sciences-National

Toxicology Program by :

A. STUDY DESIGN SUMMARY

was administered by gavage to male and female B6C3F1 mice at one of three dosages (62.5, 125, or 250 mg/kg) or a vehicle control (corn oil) group. There were 50 mice per sex in each dosage group. The first day of dosing was , for females, and

, for males. At the initiation of dosing, the females were 42 to 48 days of age and weighed 16.5 to 22.8 grams; males were 43 to 49 days of age, and weighed 20.5 to 25.5 grams.

Formulations were administered at a volume of 10 mL/kg and were based on each animal’s most recent body weight, with one exception reported…Formulations were placed on a stir plate and stirred both prior to and during administration. Dosing was completed during a consistent time period by approximately noon each day. Dosing occurred on weekdays only, exclusive of holidays, through the day prior to termination. The final day of dosing for females was

, and for males, (105 weeks of dosing for both sexes.)

B. SALIENT RESULTS AND DISCUSSION

The gavage administration of to B6C3F1 mice for up to 105 weeks resulted in decreased mean body weight and increased survival for females in the high dosage group only. -related neoplastic liver lesions (adenomas, carcinoma, and blastomas) and non-neoplastic liver foci were observed for male mice only. A decrease in pituitary gland adenomas and hyperplasias was observed for female mice only. Increases in nasal degeneration and non-neoplastic lesions of the glandular stomach were reported for both sexes.

OBSERVATIONS TO BE ADDRESSED DURING PATHOLOGY QUALITY ASSESSMENT

(OON)

The Observations to be Addressed During Pathology Quality Assessment (OON) in

Volume I, Appendix A document differences between diagnoses and accompanying organ or animal notes concerning both neoplastic and nonneoplastic lesions on PEIRPT14. The

OON also document duplicate diagnoses. All microscope slides corresponding to potential conflicts or differences in tumor diagnosis and any duplicate diagnosis were to be included in the PWG slide set and were examined during this review.

There was one potential diagnosis/organ or animal note discrepancies noted for this study.

SLIDE INVENTORY RECORDS (SIR)

The Slide Inventory Records (SIR) in Volume I, Appendix B record discrepancies between the number of slides listed on the Slide Inventory List and the number of slides actually received at . They also record the number of slides present in the slide set submitted for the current NTP review and the number of slides returned to the NTP Archives.

There were seven slide labeling discrepancies noted during this study. The labeling discrepancies are listed on the table below:

SEX/SPECIES GROUP

ANIMAL

NO.

HISTO

NO. DISCREPANCY

MALE

MICE

VM 00008 SL. NO. 7 – HISTO NO. ON GLASS READS

; PAPER LABEL READS .

SLIDE/BLOCK MATCH CONFIRMS SLIDE IS

FROM ANIMAL NO. 8.

00037 SL. NO. 8 – HISTO NO. WRITTEN ON GLASS

READS ; PAPER LABEL READS .

SLIDE/BLOCK MATCH CONFIRMS SLIDE IS

FROM ANIMAL NO. 37.

FEMALE

MICE

VF 00205 SL NO. 4 – HISTO NO. WRITTEN ON GLASS

READS ; PAPER LABEL READS

. SLIDE/BLOCK MATCH CONFIRMS

SLIDE IS FROM ANIMAL NO. 205.

MF 00310 SL. NO. 6 – HISTO NO. WRITTEN ON GLASS

READS ; PAPER LABEL READS

. SLIDE/BLOCK MATCH CONFIRMS

SLIDE IS FROM ANIMAL NO. 310.

LF 00266 SL. NO. 2 – HISTO NO. WRITTEN ON GLASS

READS ; PAPER LABEL READS .

SLIDE/BLOCK MATCH CONFIRMS SLIDE IS

FROM ANIMAL NO. 266.

00270 SL. NO. 2 – HISTO NO. READS ; PAPER

LABEL READS . SLIDE/BLOCK MATCH

CONFIRMS SLIDE IS FROM ANIMAL NO. 270.

0300 SL. NO. 6 – HISTO NO. WRITTEN ON GLASS

READS ; PAPER LABEL READS .

SLIDE/BLOCK MATCH CONFIRMS SLIDE IS

FROM ANIMAL NO. 300.

SLIDE REVIEW WORK SHEETS (SRWS)

The Slide Review Work Sheets are presented in Volume II, Appendix C (Males) and

Volume III, Appendix D (Females). These work sheets list, in animal number order, all tissues that were reviewed by a quality assessment pathologist (QAP) at . The work sheets list diagnoses included in the chronic report and comments made by the QAP

(corresponding slide numbers are listed in parentheses). Entries other than “Agree” under the reviewing pathologist’s comments indicate a disagreement with the study pathologist’s

(SP) diagnosis. In each instance, space is provided to record remarks made during the

Slide Review. The slide set contains all slides with tissues corresponding to entries on the

Slide Review Work Sheets and which were examined during this review.

A. Organs Reviewed for All Lesions

The following organs were considered to be potential target organs:

MALE MICE

Liver

Stomach, Glandular

Nose

Thyroid Gland

FEMALE MICE

Liver

Stomach, Glandular

Nose

Thyroid Gland

Ovary

B. Organs Reviewed From All Animals for Specific Lesions

The following organs (incidence organs) were examined from all animals in all groups for a designated diagnosis only:

MALE MICE

Harderian Gland – Hyperplasia

Harderian Gland – Infiltration Cellular, Mononuclear Cell

Islets, Pancreatic – Hyperplasia

Kidney – Mineralization

Kidney – Nephropathy

Spleen – Hematopoietic Cell Proliferation

Testes, Germinal Epithelium – Atrophy

Intestine, Small, All Sections – Lymphangiectasis

Lymph Node, Mesenteric – Ectasia

Liver – Fatty Change

Bone – Fibrosis

Harderian Gland – Atrophy

Harderian Gland – Hyperplasia

Kidney – Nephropathy

Pituitary Gland, Pars Distalis – Hyperplasia

Pituitary Gland, Pars Distalis – Adenoma

Spleen – Hematopoietic Cell Proliferation

Intestine, Small, All Sections – Lymphangiectasis

Lymph Node, Mesenteric – Ectasia

Liver – Fatty Change

C. Specific Lesions for Review

The following organs were reviewed only when the specific diagnosis (originally diagnosed by the SP) was present:

MALE MICE

Eye – Degeneration

Heart – Cardiomyopathy

Heart – Inflammation, Chronic

Heart, Myocardium – Mineralization

Heart, Myocardium – Necrosis

Intestine, Small, Duodenum – Polyp Adenomatous

Intestine, Small, Jejunum – Polyp Adenomatous

Kidney – Glomerulosclerosis

Kidney – Infiltration Cellular, Mononuclear Cell

MALE MICE (CONT’D)

Kidney – Inflammation, Chronic

Kidney, Renal Tubule – Hyperplasia

Oral Mucosa - Regeneration

Prostate – Infiltration Cellular, Mononuclear Cell

Prostate – Inflammation, Chronic

Seminal Vesicle – Infiltration Cellular, Mononuclear Cell

Seminal Vesicle – Inflammation, Chronic

Skin, Epidermis – Keratoacanthoma

Spleen – Atrophy

Thyroid, Follicle – Cytoplasmic Alteration

Tooth – Malformation

All Tumors in All Organs from All Animals

All diagnoses with organ notes in which either the note or the diagnosis indicates a possible discrepancy in the diagnosis of a neoplastic lesion: comment as to the appropriateness of the organ note.

All organs with possible duplicated entries as listed in the Observations to be Addressed During Pathology Quality Assessment: recommend appropriate action (for example, deletion of one entry) or add qualifier (such as; multiple, combine grades, combine TGL Numbers). Determine if lesions are multiple or different areas of the same lesion.

FEMALE MICE

Adrenal Cortex – Cytoplasmic Alteration

Eye – Inflammation, Chronic

Eye, Cornea – Inflammation, Chronic

Heart – Cardiomyopathy

Heart – Inflammation, Chronic

Heart, Myocardium – Mineralization

Lung – Hyperplasia, Lymphoid

Lung – Inflammation, Chronic

Ovary – Cytoplasmic Alteration

Salivary Glands – Infiltration Cellular

Salivary Glands – Infiltration Cellular, Mononuclear Cell

Skin – Hemangioma

Stomach, Glandular – Hyperplasia, Lymphoid

Stomach, Glandular – Inflammation, Chronic

FEMALE MICE (CONT’D)

All Tumors in All Organs from All Animals

All diagnoses with organ notes in which either the note or the diagnosis indicates a possible discrepancy in the diagnosis of a neoplastic lesion: comment as to the appropriateness of the organ note.

All organs with possible duplicated entries as listed in the Observations to be Addressed During Pathology Quality Assessment: recommend appropriate action (for example, deletion of one entry) or add qualifier (such as; multiple, combine grades, combine TGL Numbers). Determine if lesions are multiple or different areas of the same lesion.

FINDINGS OF THE SLIDE REVIEW

The slides reviewed during this quality assessment were of good quality and had no artifacts that interfered with making diagnoses.

Neoplastic to Nonneoplastic Differences There were several neoplasms listed in PEIRPT14 that the QAP considered to be nonneoplastic lesions and they are as follows:

NEOPLASTIC TO NONNEOPLASTIC DIFFERENCES

SEX GROUP

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT1 QAP COMMENTS

MALE 0 MG/KG 18 LIVER HEPATOCLR ADEN EOSIN FOCUS

20 LIVER HEPATOCLR ADEN EOSIN FOCUS

25 STOM FORESTOM SQ CL PAPILLOM

EPITHELIUM –

HYPERPLASIA

26 LIVER HEPATOCLR ADEN MIXED CL FOCUS

THYROID GL,

FOLLICULAR CEL ADENOMA FOLLICLE – DEGEN

34 LIVER HEPATOCLR ADEN MIXED CELL FOCUS

34 THYMUS THYMOMA BGN

EPITHELIAL CELL –

HYPERPLASIA

62.5

MG/KG 59 LIVER HEPATOCLR ADEN EOSIN FOCUS

NEOPLASTIC TO NONNEOPLASTIC DIFFERENCES

SEX GROUP

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT14 QAP COMMENTS

MALE

62.5

MG/KG 63 LIVER HEPATOCLR ADEN BASOPH FOCUS

LYMPH NODE,

PANCREATIC LYMPH MAL NECROSIS

69 STOM FORESTOM SQ CL PAPILLOM

EPITHELIUM –

HYPERPLASIA

70 LIVER HEPATOCLR ADEN EOSIN FOCUS

71 BONE MARROW HISTIO SARC

AUTOLYSIS

PRECLUDES

EXAMINATION

LIVER HISTIO SARC

AUTOLYSIS

PRECLUDES

EXAMINATION

SKIN, SEBACEOUS

GL CARCINOMA

HARDERIAN GL –

INFLAMMATION,

CHRONIC

99 LIVER HEPTOCLR ADEN EOSIN FOCUS

MG/KG 103 LIVER HEPATOCLR ADEN EOSIN FOCUS

104 SKIN, EPIDERMIS KERATOACANTHMA

INSUFFICIENT

TISSUE

123 LIVER HEPATOCLR ADEN EOSIN FOCUS

130 SPLEEN HEMANGIOSARC

AUTOLYSIS

PRECLUDES

EXAMINATION

MG/KG 169

THRYOID GL,

FOLLICULAR CEL CARCINOMA

THYROID GL –

INFLAMMATION,

CHRONIC

184 EPIDIDYMIS HEMANGIOMA ANGIECTASIS

LIVER

HEPATOCLR ADEN,

MULTIPLE EOSIN FOCUS

200 LIVER

HEPATOCLR ADEN,

MULTIPLE EOSIN FOCUS

FEMALE 0 MG/KG 201

THYROID GL,

FOLLICULAR CEL ADENOMA

NOT PRESENT IN

SECTION

PITUITARY GL, PARS

DISTALIS ADENOMA HYPERPLASIA

212 OVARY CYSTADENOMA CYST

NEOPLASTIC TO NONNEOPLASTIC DIFFERENCES

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT14 QAP COMMENTS

FEMALE 0 MG/KG 217 SPLEEN HEMANGIOSARC ANGIECTASIS

PITUITARY GL, PARS

INTERMED ADENOMA HYPERPLASIA

PITUITARY GL, PARS

DISTALIS ADENOMA

INSUFFICIENT

TISSUE

231 OVARY CYSTADENOMA

NOT PRESENT IN

SECTION

242 ADREN MEDULLA PHEOCHROM BGN HYPERPLASIA

249 INT LG CECUM LYMPH MAL

LYMPHOID TISSUE –

HYPERPLASIA

IN SM ILEUM LYMPH MAL

LYMPHOID TISSUE –

HYPERPLASIA

62.5

MG/KG 269 LUNG

HEPATOCLR CARC,

METASTATIC

NOT PRESENT IN

SECTION

KIDNEY, RENAL

TUBULE ADENOMA

NO SECTION

PRESENT

PITUITARY GL, PARS

DISTALIS ADENOMA HYPERPLASIA

MG/KG 309 ADREN MEDULLA PHEOCHROM BGN HYPERPLASIA

336 PARATHYROID GL ADENOMA

NOT PRESENT IN

SECTION

346 HARDERIAN GL CARCINOMA HYPERPLASIA

MG/KG 353 SPLEEN HISTIO SARC

HYPERPLASIA, MAST

CELL

Nonneoplastic to Neoplastic Differences There were several neoplasms observed during this review which either had not been diagnosed previously or had been considered nonneoplastic lesions by the SP, and they are as follows:

NONNEOPLASTIC TO NEOPLASTIC DIFFERENCES

SEX GROUP

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT14 QAP COMMENTS

MALE 0 MG/KG 39 LIVER -

HEPATOCLR ADEN,

MULTIPLE

NONNEOPLASTIC TO NEOPLASTIC DIFFERENCES

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT14 QAP COMMENTS

MALE 0 MG/KG 41 KIDNEY -

HEPATOCLR CARC,

METASTATIC

62.5

MG/KG 91 LIVER - HEPATOCLR ADEN

93 LIVER -

HEPATOCLR ADEN,

MULTIPLE

MG/KG 111

HARDERIAN

GL - ADENOMA

HARDERIAN

GL HYPERPLASIA ADENOMA

131 LIVER - HEPATOCLR ADEN

136 STOM GLAND NORMAL

CARCINOMA,

METASTATIC

137 LIVER - HEPATOCLR ADEN

140 LUNG - ALV BRON ADEN

141 LIVER MIXED CL FOCUS

HEPATOCLR ADEN,

MULTIPLE

MG/KG 151

TESTES,

INTERSTIT CEL - ADENOMA

155 STOM GLAND -

CARCINOMA,

METASTATIC

158 LIVER - HEMANGIOSARC

ISLETS PANC HYPERPLASIA ADENOMA

ADREN

CORTEX,

SUBCAPSULAR - ADENOMA

LIVER MIXED CL FOCUS HEPATOCLR ADEN

FEMALE 0 MG/KG 215 LIVER MIXED CL FOCUS HEPATOCLR ADEN

INT SM

DUODEN NORMAL ADENOMA

INT SM

JEJUNUM - LEIOMYOSAR

ADREN

CORTEX,

SUBCAPSULAR - ADENOMA

LIVER BASOPH FOCUS HEPATOCLR ADEN

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT14 QAP COMMENTS

FEMALE 0 MG/KG 248

OVARY,

CORPUS

LUTEUM HYPERPLASIA

OVARY – GRANLSA

TM BGN

ADREN

CORTEX - ADENOMA

62.5

MG/KG 258 LIVER MIXED CL FOCUS HEPATOCLR ADEN

265 LIVER - LYMPH MAL

KIDNEY NORMAL LYMPH MAL

SPLEEN HEMA CELL PROL LYMPH MAL

266 LIVER - LYMPH MAL

LUNG - LYMPH MAL

LYM NODE

MESEN - LYMPH MAL

KIDNEY NORMAL LYMPH MAL

SPLEEN NORMAL LYMPH MAL

HARDERIAN

GL - ADENOMA

LIVER - HEPATOCLR ADEN

279 SPLEEN HEMA CELL PROL LYMPH MAL

LYMPH NODE

MESEN NORMAL LYMPH MAL

MG/KG 313

HARDERIAN

GL HYPERPLASIA ADENOMA

342 NOSE - LYMPH MAL

BONE NORMAL LYMPH MAL

342 STOM GLAND

INFLAMMATION,

CHRONIC LYMPH MAL

MG/KG 351 UTERUS - POLYP STROMAL

352 UTERUS - POLYP STROMAL

ANIMAL

NO. TISSUE

DIAGNOSIS ON

PEIRPT14 QAP COMMENTS

FEMALE

MG/KG 353 UTERUS - POLYP STROMAL

397 UTERUS - POLYP STROMAL

399 OVARY ANGIECTASIS HEMANGIOSARC

Target Organs Glandular Stomach

The glandular stomach was considered a target organ in both male and female mice due to increases in the incidences of stomach, glandular – inflammation, chronic;

stomach, glandular – hyperplasia; and stomach, glandular – necrosis. Generally, this review corroborated the incidence of stomach, glandular – inflammation, chronic and stomach, glandular – hyperplasia. However, the diagnosis of stomach, glandular – necrosis was not verified by this review.

In the final report, stomach, glandular – hyperplasia was characterized as increased basophilia of epithelium and increased numbers of mucus neck cells by the study pathologist. Generally, gastric glands were increased in length (25%-50%) with occasional dilation and distortion of gastric pits. There appeared to be greater numbers of parietal cells;

chief cells were slightly more frequent in some areas. Glandular hyperplasia was accompanied by stomach, glandular – inflammation, chronic that was described by the study pathologist as increased mononuclear and polymorphonuclear inflammatory cells and increased pigment-containing macrophages. There were minimal to mild infiltrates of lymphocytes, macrophages, and infrequent neutrophils adjacent to the muscularis mucosae with extension into the overlying lamina propria. Many infiltrating histiocytic cells contained intracytoplasmic, light-brown granular pigment that was recorded separately by the QAP as stomach, glandular – pigmentation. Focal aggregates of pigment were frequently observed within gastric pits. Evidence of test-article related stomach, glandular – necrosis was described by the SP as degenerative cytologic changes in or absence of epithelium, dilatation of gastric pits or pigment in the lumen of gastric pits. The QAP did not observe notable gastric epithelial degeneration nor regarded gastric pit dilatation and pigment aggregation as components of a necrotizing process.

The incidences of selected lesions in the glandular stomach diagnosed by the SP and recorded during this review (QAP) are listed in the following tables:

INCIDENCES OF SELECTED LESIONS IN THE GLANDULAR STOMACH

MALE MICE

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Natural Death 1 - - - Survivors – Terminal Sacrifice 26 31 32 32

SP QAP SP QAP SP QAP SP QAP

STOMACH, GLANDULAR (NO. EX.) (50) (50) (50) (47) (50) (47) (50) (49)

INFLAMMATION, CHRONIC 1 1 0 1 11 18 37 45

MINIMAL 1 1 0 1 11 18 35 43

MILD 0 0 0 0 0 0 2 2

PIGMENTATION 0 0 0 1 0 38 0 48

MINIMAL 0 0 0 1 0 38 0 45

MILD 0 0 0 0 0 0 0 3

EPITHELIUM

ATROPHY 0 0 0 0 0 0 0 1

MODERATE 0 0 0 0 0 0 0 1

HYPERPLASIA 0 0 1 0 27 20 47 38

MINIMAL 0 0 1 0 23 16 24 18

MILD 0 0 0 0 4 4 23 20

NECROSIS 0 0 1 0 24 0 44 0

MINIMAL 0 0 1 0 24 0 42 0

MILD 0 0 0 0 0 0 2 0

INCIDENCES OF SELECTED LESIONS GLANDULAR STOMACH

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

STOMACH, GLANDULAR (NO. EX.) (50) (48) (50) (50) (50) (49) (50) (50)

INFLAMMATION, CHRONIC 0 0 6 15 14 28 44 47

MINIMAL 0 0 6 15 13 26 31 33

MILD 0 0 0 0 1 2 13 14

PIGMENTATION 0 0 0 16 0 31 0 49

MINIMAL 0 0 0 16 0 24 0 6

MILD 0 0 0 0 0 7 0 43

EPITHELIUM

HYPERPLASIA 1 0 5 7 8 10 35 35

MINIMAL 0 0 3 5 8 8 22 22

MILD 1 0 2 2 0 2 13 13

NECROSIS 0 0 7 0 16 0 46 0

MINIMAL 0 0 7 0 16 0 43 0

MILD 0 0 0 0 0 0 3 0

Liver The liver was considered a target organ in male mice due to increases in the incidences of liver – clear cell focus; liver – eosinophilic focus; liver – fatty change;

liver- mixed cell focus; liver – hepatoblastoma; liver – hepatoblastoma, multiple; liver

– hepatocellular adenoma; liver – hepatocellular adenoma, multiple; liver – hepatocellular carcinoma; and liver – hepatocellular carcinoma, multiple. In female mice, the liver was a target organ due to increases in the incidences of liver – hepatoblastoma and liver – hepatocellular adenoma.

In male mice, the QAP generally diagnosed liver – hepatocellular carcinoma less frequently than the SP. The QAP frequently modified the morphologic diagnosis from liver – hepatocellular carcinoma to liver – hepatocellular adenoma. Consequently, when compared to the SP’s findings there were decreases in the incidences of liver – hepatocellular carcinoma, multiple with concurrent increases in the incidences of liver – hepatocellular adenoma, multiple. Generally, this review concurred with the SP’s diagnosis of liver – eosinophilic focus; however, many cases of liver – hepatocellular adenoma were downgraded to liver – eosinophilic focus. Consequently, this review resulted in nominal increases in the incidences of liver-eosinophilic focus in male mice.

Furthermore, this review generally supported the SP’s diagnosis of liver – clear cell focus;

liver – hepatoblastoma; and liver – hepatoblastoma, multiple in male mice. However, the QAP diagnosed liver – basophilic focus slightly more frequently as compared to the

SP. Moreover, the QAP accepted the SP’s finding of liver – fatty change, however it was not considered to be a significant test-article effect.

In female mice, the QAP frequently modified the SP’s diagnoses of liver – hepatocellular carcinoma, multiple to simply liver – hepatocellular carcinoma. In several instances of multiple carcinomas, the QAP disagreed with histomorphologic criteria used in the SP’s assessment of one or more of the tumors resulting in some carcinomas being downgraded to adenomas. Consequently, there were slight increases in the incidences of liver – hepatocellular carcinoma as well as liver – hepatocellular adenoma, multiple. Similar to male mice, the QAP concurred with the SP’s diagnosis of liver –fatty change but did not consider it to be a significant test-article effect. Furthermore, the QAP did not corroborate the SP’s diagnoses of liver – clear cell focus. The QAP considered the putative clear cell foci to be focal regions of fatty change. Moreover, this review generally verified the incidence of liver – eosinophilic focus; however, the QAP corroborated the diagnosis of liver – mixed cell focus less frequently than the SP.

This review did not support the diagnosis of liver – tension lipidosis in either male or female mice. Tension lipidosis is not considered to be a valid pathologic entity and is devoid of functional significance.

The incidences of selected lesions in the liver diagnosed by the SP and recorded during this review (QAP) are listed in the following tables:

INCIDENCES OF SELECTED LESIONS IN THE LIVER

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Natural Death 1 - - - Survivors – Terminal Sacrifice 26 31 32 32

SP QAP SP QAP SP QAP SP QAP

LIVER (NO. EX.) (50) (50) (50) (49) (50) (49) (50) (50)

BASOPHILIC FOCUS 1 2 3 4 2 2 4 6

CLEAR CELL FOCUS 7 6 17 18 17 21 22 19

EOSINOPHILIC FOCUS 24 27 35 36 31 37 31 34

FATTY CHANGE 29 29 26 27 28 28 37 36

MINIMAL 8 8 15 15 15 15 16 16

MILD 21 21 11 11 13 13 21 20

MODERATE 0 0 0 1 0 0 0 0

HEMANGIOMA 0 0 0 0 0 0 1 1

HEMANGIOSARCOMA 3 3 2 2 1 1 2 2

HEMANGIOSARCOMA, MULTIPLE 1 1 2 2 0 0 1 1

HEPATOBLASTOMA 3 3 3 3 1 1 6 7

HEPATOBLASTOMA, MULTIPLE 0 0 1 1 3 3 7 7

HEPATOCELLULAR ADENOMA 13 9 12 7 12 11 8 11

HEPATOCELLULAR ADENOMA, MULTIPLE 15 16 16 26 12 21 30 32

HEPATOCELLULAR CARCINOMA 10 7 15 5 13 9 10 7

HEPATOCELLULAR CARCINOMA, MULTIPLE 5 4 5 3 23 14 18 6

HEPATOCHOLANGIOCARCINOMA 0 0 3 3 1 0 1 0

MIXED CELL FOCUS 1 2 10 3 12 4 11 3

INCIDENCES OF SELECTED LESIONS LIVER

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

LIVER (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

BASOPHILIC FOCUS 4 4 5 6 6 6 2 5

CLEAR CELL FOCUS 4 0 2 0 2 0 1 0

EOSINOPHILIC FOCUS 15 15 22 26 21 26 20 21

FATTY CHANGE 36 36 39 39 35 35 40 40

MINIMAL 16 16 21 21 20 20 17 17

MILD 16 17 17 17 12 12 23 23

MODERATE 2 1 1 1 2 2 0 0

MARKED 2 2 0 0 1 1 0 0

INCIDENCES OF SELECTED LESIONS LIVER

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

LIVER (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

HEMANGIOSARCOMA, MULTIPLE 0 0 0 0 1 1 0 0

HEPATOBLASTOMA 0 0 0 0 1 1 1 1

HEPATOCELLULAR ADENOMA 5 8 11 14 7 6 9 7

HEPATOCELLULAR ADENOMA, MULTIPLE 1 0 2 3 0 2 3 4

HEPATOCELLULAR CARCINOMA 6 6 7 6 7 9 3 4

HEPATOCELLULAR CARCINOMA, MULTIPLE 0 0 2 1 3 0 1 0

HEPATOCHOLANGIOCARCINOMA 0 0 1 0 0 0 0 0

MIXED CELL FOCUS 4 1 10 1 10 3 5 0

Nose The nose was considered a target organ in both male and female mice due to increases in the incidences of nose, glands, respiratory epithelium – hyperplasia; nose, nerve – atrophy; and nose, olfactory epithelium – degeneration. There was an increased incidence of nose – inflammation in female mice only. Furthermore, the QAP added the diagnoses of nose, respiratory epithelium – accumulation, hyaline droplet and nose, olfactory epithelium – accumulation, hyaline droplet throughout the study.

Generally, this review did not support the SP’s diagnosis of nose, olfactory epithelium – degeneration. In the final report, the SP described nose, olfactory epithelium – degeneration as being characterized histologically by decreased numbers of olfactory epithelial cells and/or replacement of olfactory epithelium with glandular or respiratory epithelium. These changes were consistent with nose, olfactory epithelium – respiratory metaplasia and the QAP chose to modify the SP’s diagnosis accordingly.

Mild to moderate respiratory metaplasia of olfactory epithelium was commonly observed at

Levels II and III in both males and females at the 125 mg/kg and 250 mg/kg dose levels.

Minimal respiratory metaplasia was infrequently observed at Level III in lower dose groups and untreated controls. In markedly affected sections, the metaplastic change was characterized by focally extensive to diffuse replacement of psuedostratified columnar olfactory epithelium with ciliated cuboidal to columnar respiratory epithelium that frequently extended into the subjacent lamina propria and lined Bowman’s glands. Bowman’s glands were frequently dilated and tortuous. The NTP describes olfactory epithelium – degeneration as being characterized by the presence of one or more of the following:

epithelial vacuolization, bleb formation, cellular disorientation, cytoplasmic eosinophilia, cell swelling, and/or loss of cell to cell contact. In a few instances, the QAP supported the SP’s diagnosis of degeneration when one or more of the aforementioned changes were observed in olfactory epithelium. In some cases, respiratory metaplasia was accompanied by concomitant incidences of nose, olfactory epithelium – atrophy exemplified by thinning of olfactory epithelium as a consequence of cell loss. Furthermore, in many cases the severity of the respiratory metaplasia resulted in atrophy of axonal nerve bundles. Nose, nerve – atrophy was characterized by the presence of shrunken olfactory nerve bundles in the lamina propria at Levels II and III. This review concurred with the SP’s findings.

Generally, this review supported the SP’s finding of respiratory epithelial hyperplasia, however, the QAP chose to modify the morphologic diagnosis from nose, glands, respiratory epithelium – hyperplasia to nose, respiratory epithelium – hyperplasia in an effort to sustain consistency with NTP nomenclature. Nose, respiratory epithelium – hyperplasia was typically observed at Level II with slightly less frequency at Level I and was characterized by proliferation of ciliated columnar cells that occasionally formed villous structures that projected into nasal passages. Frequently, these cells extended into the glands of the subjacent lamina propria that were often dilated and hyperplastic. Hyperplasia of the respiratory epithelium of both the nasal mucosa and submucosal glands were captured in the singular diagnosis of nose, respiratory epithelium – hyperplasia.

Consequently, the QAP diagnosed respiratory epithelium hyperplasia significantly more frequently than the SP in males and females.

Nose, respiratory epithelium – accumulation, hyaline droplet and nose, olfactory epithelium – accumulation, hyaline droplet were observed in sections of nose from treated males and females as well as untreated controls. Hyaline droplet accumulation was demonstrated by the intracytoplasmic presence of homogeneous, eosinophilic, hyaline globules that was commonly observed in both the respiratory and olfactory epithelium at

Level III as well as the respiratory epithelium lining the nasopharyngeal duct.

Furthermore, the QAP diagnosed nose, olfactory epithelium – necrosis and nose, respiratory epithelium – necrosis in a few males of the 250 mg/kg dose group. The QAP also diagnosed nose, olfactory epithelium – necrosis in two females of the 125 mg/kg dose group. The QAP added the diagnosis of nose, olfactory epithelium – metaplasia, squamous (Level III) in two males of the 250 mg/kg dose group. Furthermore, the diagnosis of nose, respiratory epithelium – metaplasia, squamous was added in a single high dose male as well as an untreated control female.

This review concurred with the SP’s diagnoses of nose – inflammation. In the final report the SP described inflammation of the nasal passages as being characterized by protein accumulations and/or infiltration of neutrophils and mononuclear inflammatory cells in the lumen. Inflammation was generally minimal and not a prominent feature.

The incidences of selected lesions in the nose diagnosed by the SP and recorded during this review (QAP) are listed in the following tables:

INCIDENCES OF SELECTED LESIONS IN THE NOSE

62.5

MG/KG

MG/KG

MG/KG

Survivors – Natural Death 1 - - - Survivors – Terminal Sacrifice 26 31 32 32

SP QAP SP QAP SP QAP SP QAP

NOSE (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

INFLAMMATION 16 13 12 11 12 12 14 16

MINIMAL 14 11 10 9 11 11 10 11

MILD 2 2 2 2 1 1 3 4

MODERATE 0 0 0 0 0 0 1 1

POLYP, INFLAMMATORY 0 1 1 2 3 3 1 1

MINIMAL 0 0 1 1 3 3 0 0

MILD 0 1 0 1 0 0 1 1

GLANDS, RESPIRATORY EPITHELIUM

HYPERPLASIA 13 0 18 0 25 0 28 0

MINIMAL 13 0 18 0 20 0 13 0

MILD 0 0 0 0 5 0 9 0

MODERATE 0 0 0 0 0 0 6 0

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Natural Death 1 - - - Survivors – Terminal Sacrifice 26 31 32 32

SP QAP SP QAP SP QAP SP QAP

NOSE (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

NERVE

ATROPHY 0 0 0 0 0 0 7 8

MINIMAL 0 0 0 0 0 0 1 1

MILD 0 0 0 0 0 0 6 6

MODERATE 0 0 0 0 0 0 0 1

OLFACTORY EPITHELIUM

ACCUMULATION, HYALINE DROPLET 0 4 0 4 0 5 0 5

MINIMAL 0 4 0 3 0 5 0 5

MILD 0 0 0 1 0 0 0 0

ATROPHY 0 3 0 4 0 10 0 17

MINIMAL 0 2 0 1 0 7 0 12

MILD 0 1 0 3 0 2 0 1

MODERATE 0 0 0 0 0 1 0 4

DEGENERATION 13 0 19 0 25 3 29 1

MINIMAL 13 0 15 0 15 2 13 0

MILD 0 0 4 0 9 1 9 1

MODERATE 0 0 0 0 1 0 7 0

METAPLASIA, SQUAMOUS 0 0 0 0 0 0 0 2

MILD 0 0 0 0 0 0 0 1

MODERATE 0 0 0 0 0 0 0 1

NECROSIS 0 0 0 0 0 0 0 5

MINIMAL 0 0 0 0 0 0 0 2

MILD 0 0 0 0 0 0 0 2

MARKED 0 0 0 0 0 0 0 1

RESPIRATORY METAPLASIA 0 14 0 14 0 20 0 28

MINIMAL 0 13 0 10 0 12 0 19

MILD 0 1 0 4 0 7 0 6

MODERATE 0 0 0 0 0 1 0 3

RESPIRATORY EPITHELIUM

ACCUMULATION, HYALINE DROPLET 0 18 0 34 0 30 0 26

MINIMAL 0 15 0 30 0 28 0 22

MILD 0 3 0 3 0 2 0 4

MODERATE 0 0 0 1 0 0 0 0

HYPERPLASIA 0 36 0 40 0 41 0 45

MINIMAL 0 36 0 34 0 31 0 35

MILD 0 0 0 6 0 10 0 6

MODERATE 0 0 0 0 0 0 0 4

METAPLASIA, SQUAMOUS 0 0 0 0 0 0 0 1

MILD 0 0 0 0 0 0 0 1

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Natural Death 1 - - - Survivors – Terminal Sacrifice 26 31 32 32

SP QAP SP QAP SP QAP SP QAP

NOSE (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

RESPIRATORY EPITHELIUM (CONT’D)

NECROSIS 0 0 0 0 0 0 0 2

MINIMAL 0 0 0 0 0 0 0 1

MILD 0 0 0 0 0 0 0 1

INCIDENCES OF SELECTED LESIONS NOSE

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

NOSE (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

INFLAMMATION 4 4 1 1 8 8 39 39

MINIMAL 2 2 1 1 7 7 30 30

MILD 2 2 0 0 1 1 9 9

GLANDS, RESPIRATORY EPITHELIUM

HYPERPLASIA 13 0 11 0 23 0 50 0

MINIMAL 13 0 11 0 21 0 0 0

MILD 0 0 0 0 1 0 0 0

MODERATE 0 0 0 0 1 0 50 0

NERVE

ATROPHY 0 0 0 0 1 1 50 50

MILD 0 0 0 0 1 1 2 2

MODERATE 0 0 0 0 0 0 48 48

OLFACTORY EPITHELIUM

ACCUMULATION, HYALINE DROPLET 0 17 0 27 0 21 0 44

MINIMAL 0 15 0 26 0 18 0 1

MILD 0 1 0 1 0 3 0 39

MODERATE 0 1 0 0 0 0 0 3

MARKED 0 0 0 0 0 0 0 1

ATROPHY 0 1 0 2 0 3 0 45

MINIMAL 0 1 0 2 0 1 0 2

MILD 0 0 0 0 0 1 0 41

MODERATE 0 0 0 0 0 1 0 2

INCIDENCES OF SELECTED LESIONS NOSE

MG/KG

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

NOSE (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

OLFACTORY EPITHELIUM (CONT’D)

DEGENERATION 13 0 11 0 23 0 50 2

MINIMAL 13 0 11 0 20 0 0 0

MILD 0 0 0 0 2 0 0 0

MODERATE 0 0 0 0 1 0 50 2

NECROSIS 0 0 0 0 0 2 0 0

MINIMAL 0 0 0 0 0 1 0 0

MILD 0 0 0 0 0 1 0 0

RESPIRATORY METAPLASIA 0 7 0 8 0 17 0 49

MINIMAL 0 7 0 8 0 16 0 1

MILD 0 0 0 0 0 0 0 1

MODERATE 0 0 0 0 0 1 0 47

RESPIRATORY EPITHELIUM

ACCUMULATION, HYALINE DROPLET 0 47 0 38 0 44 0 50

MINIMAL 0 31 0 36 0 41 0 1

MILD 0 15 0 2 0 3 0 29

MODERATE 0 1 0 0 0 0 0 19

MARKED 0 0 0 0 0 0 0 1

HYPERPLASIA 0 32 0 30 0 38 0 50

MINIMAL 0 31 0 30 0 37 0 0

MILD 0 1 0 0 0 1 0 2

MODERATE 0 0 0 0 0 0 0 48

METAPLASIA, SQUAMOUS 0 1 0 0 0 0 0 0

MINIMAL 0 1 0 0 0 0 0 0

NECROSIS 0 2 0 0 0 1 0 0

MINIMAL 0 2 0 0 0 1 0 0

Thyroid Gland As a consequence of lesions observed with related compounds, the thyroid gland was considered a target organ and reviewed for proliferative follicular effects (i.e., hypertrophy, hyperplasia, adenoma, carcinoma). There were no definitive test article effects on the thyroid gland of either male or female mice observed during this review. The QAP noted thyroid gland – degeneration in both a single control, as well as high dose male mouse. Degeneration is a common age-related change of the murine thyroid gland.

The incidences of selected lesions in the thyroid gland diagnosed by the SP and recorded during this review (QAP) are listed in the following tables:

INCIDENCES OF SELECTED LESIONS IN THE THYROID GLAND

62.5

MG/KG

MG/KG

MG/KG

Survivors – Natural Death 1 - - - Survivors – Terminal Sacrifice 26 31 32 32

SP QAP SP QAP SP QAP SP QAP

THYROID GLAND (NO. EX.) (50) (50) (50) (50) (50) (49) (50) (50)

C CELL

ADENOMA 0 0 1 1 0 0 1 1

FOLLICLE

CYTOPLASMIC ALTERATION 0 0 0 0 0 0 1 0

MILD 0 0 0 0 0 0 1 0

DEGENERATION 0 1 0 0 0 0 0 1

MILD 0 1 0 0 0 0 0 1

FOLLICULAR CELL

ADENOMA 1 0 0 0 1 1 1 1

CARCINOMA 0 0 0 0 0 0 1 0

INCIDENCES OF SELECTED LESIONS THYROID GLAND

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

THYROID GLAND (NO. EX.) (50) (50) (50) (50) (50) (49) (50) (50)

C CELL

CARCINOMA 0 1 0 0 0 0 0 0

FOLLICLE

HYPERPLASIA 0 0 0 0 0 0 1 1

MILD 0 0 0 0 0 0 1 1

FOLLICULAR CELL

ADENOMA 2 1 0 0 0 0 1 1

CARCINOMA 1 0 0 0 0 0 0 0

Ovary The ovary was considered to be a target organ in female mice due to decreases in the incidences of ovary – cyst. Generally, this review supported the SP’s findings. Ovarian cysts are a common age-related change in female mice.

The incidences of selected lesions in the ovary diagnosed by the SP and recorded during this review (QAP) are listed in the following tables:

INCIDENCES OF SELECTED LESIONS OVARY

62.5

MG/KG

MG/KG

MG/KG

Survivors – Moribund Sacrifice - - 1 - Survivors – Terminal Sacrifice 33 40 25 45

SP QAP SP QAP SP QAP SP QAP

OVARY (NO. EX.) (50) (50) (50) (50) (50) (50) (50) (50)

CYST 17 15 15 15 10 9 8 8

MINIMAL 10 7 9 9 4 3 6 6

MILD 7 8 6 6 6 6 2 2

CYSTADENOMA 3 1 4 4 1 1 4 4

OTHER REVIEWED ORGANS/LESIONS

All sections of spleen were reviewed in both male and female mice for the incidence of spleen – hematopoietic cell proliferation. Hematopoietic cell proliferation is a normal process in the mouse spleen and should not be recorded as a pathologic change unless the level is above background. However, the QAP experienced difficulty in ascertaining the SP’s threshold for recording this finding. Consequently, the QAP chose to generally support the

SP’s findings in spite of its insignificance to the study.

All sections of Harderian gland were reviewed in male mice for Harderian gland – hyperplasia as well as Harderian gland – infiltration cellular, mononuclear cell and in female mice for Harderian gland – hyperplasia and Harderian gland – atrophy. This review generally confirmed the incidence of Harderian gland – hyperplasia as defined by the SP in both male and female mice. Hyperplasia was typified by focal areas of increased numbers of slightly enlarged alveolar cells in the absence of clearly defined compression of the surrounding alveoli. Hyperplastic cells were slightly more basophilic than unaffected cells. The QAP considered the incidence and severity of Harderian gland – infiltration cellular, mononuclear cell to be consistent with minimal chronic inflammation commonly identified in aged mice as a spontaneous lesion. The QAP recorded these changes as “not remarkable”. This review did not concur with the SP’s diagnosis of Harderian gland – atrophy in female mice. The QAP assessed these changes to be consistent with dilatation of glandular alveoli and appropriately modified the diagnosis to Harderian gland – dilatation.

All sections of kidney were reviewed in male mice for kidney – mineralization, as well as kidney – nephropathy and in female mice for kidney – nephropathy only. The

QAP diagnosed kidney – nephropathy at a slightly greater frequency than the SP in both male and female mice. Moreover, the QAP recorded most of the SP’s diagnoses of kidney

– mineralization in male mice as “not remarkable”. In most sections, the severity of mineralization was minimal as evidenced by the presence of one or two small foci of mineralization involving renal tubules or basement membranes. The SP inconsistently recognized and diagnosed these regions of minimal mineralization; consequently, the QAP verified the finding only when the severity was above background. Furthermore, in male mice the specific diagnoses of kidney – glomerulosclerosis; kidney – infiltration cellular, mononuclear cell; kidney – inflammation, chronic; and kidney, renal tubule – hyperplasia were reviewed. The QAP did not verify the SP’s diagnoses of kidney – glomerulosclerosis and kidney, renal tubule – hyperplasia (as recorded in an untreated control mouse). However, the QAP added the diagnosis of kidney, renal tubule – hyperplasia in a single male of the 125 mg/kg dose group. The SP’s diagnosis of kidney – infiltration cellular in a single untreated control mouse was modified by the QAP to kidney

– inflammation, chronic to maintain consistency.

All sections of pancreas were reviewed in male mice for islets, pancreatic – hyperplasia only. Given the tremendous variation in the size of murine pancreatic islets the

QAP elected not to corroborate nearly all of the SP’s diagnoses of islets, pancreatic – hyperplasia. In this study, the QAP considered the size of the pancreatic islets to be well within the normal limits of variation. However, the QAP concurred with the SP’s diagnosis of islets, pancreatic – hyperplasia in a single male of the 125 mg/kg dose group.

Furthermore, this review verified a single incidence of islets, pancreatic – adenoma in an untreated male as well as an additional finding was recorded by the QAP in a single male of the 250 mg/kg dose group.

All sections of testes were reviewed in male mice for testes, germinal epithelium – atrophy. This review concurred with the SP’s diagnoses in the 250 mg/kg dose group;

however, the QAP recorded additional findings in two untreated controls as well as three mice in the 125 mg/kg dose group.

The pituitary glands of all female mice were reviewed for pituitary gland, pars distalis – hyperplasia and pituitary gland, pars distalis – adenoma. Generally, the QAP diagnosed pituitary gland, pars distalis – hyperplasia less frequently than the SP. There were several presentations of “hyperplasia” that the QAP considered to be unconvincing.

Consequently, the QAP recorded these findings as “not remarkable”. Also, there were a few examples of pituitary gland, pars distalis – adenoma that were downgraded to pituitary gland, pars distalis – hyperplasia by the QAP.

All sections of the small intestine in both male and female mice were reviewed for lymphangiectasis due to the presence of this lesion in the companion study. In male mice, the QAP diagnosed intestine small, duodenum – lymphangiectasis in a single untreated control as well as a single mouse in the 125 mg/kg dose group and two mice of the 250 mg/kg dose group. Lymphangiectasis was not diagnosed in any section of the small intestine in female mice.

All sections of mesenteric lymph nodes of both male and female mice were reviewed for lymph node, mesenteric – ectasia. Ectasia of the mesenteric lymph node was not observed during the course of this review.

All sections of bone in female mice were reviewed for the incidence of bone – fibrosis. As suggested in the Pathology Data Review, the QAP modified the diagnosis to bone – fibro-osseous lesion. Moreover, the QAP recorded the diagnosis in several additional sections from treated mice, as well as untreated controls.

The QAP modified the SP’s diagnoses of eye – degeneration to eye – phthisis bulbi in a single untreated control male mouse and a single male of the 62.5 mg/kg dose group.

In male mice, the QAP did not corroborate the SP’s diagnosis of heart, myocardium – necrosis. The SP’s diagnoses of heart – cardiomyopathy; heart – inflammation, chronic;

and heart, myocardium – mineralization were generally verified by this review.

In female mice, this review generally confirmed the incidence of heart – cardiomyopathy but not the diagnosis of heart – inflammation, chronic in a single female of the 62.5 mg/kg dose group. This diagnosis was modified to heart – cardiomyopathy. This review verified the incidences of heart, myocardium – mineralization in two control females, as well as a single female of the 250 mg/kg dose group.

All diagnoses of intestine small, duodenum – polyp adenomatous and intestine small, jejunum – polyp adenomatous in male mice were modified to intestine small, duodenum – adenoma and intestine small, jejunum – adenoma by the QAP. Furthermore, the QAP diagnosed intestine small, duodenum – adenoma in a single untreated control male.

This review did not corroborate the diagnosis of oral mucosa – regeneration in a single male of the 250 mg/kg dose group.

In male mice, the specific diagnoses of prostate – infiltration cellular, mononuclear cell; prostate – inflammation, chronic; seminal vesicle – infiltration cellular, mononuclear cell; and seminal vesicle – inflammation, chronic were reviewed. The SP inconsistently recorded these findings. Furthermore, the QAP considered the incidence and severity of the changes to be common background findings and (consequently) recorded them as “not remarkable”.

Moreover, in male mice the specific diagnoses of skin, epidermis – keratoacanthoma; spleen – atrophy; and tooth – malformation were reviewed. The single incident of skin, epidermis – keratoacanthoma in a mouse of the 125 mg/kg dose group was not verified by the QAP due to the poor quality of the section and was appropriately recorded as “insufficient tissue”. The diagnosis of spleen – atrophy was modified to spleen, lymphoid follicle – atrophy when verified by the QAP. All diagnoses of tooth – malformation were modified to tooth – dysplasia by the QAP. Furthermore, the QAP recorded minimal dental dysplasia at a greater frequency as compared to the SP. Minimal dysplasia was recorded based on the presence of mature and immature intrapulpal denticles.

In female mice, the specific diagnoses of adrenal cortex – cytoplasmic alteration;

ovary – cytoplasmic alteration; eye – inflammation, chronic; and eye, cornea – inflammation, chronic were reviewed. This review did not support the diagnoses of adrenal cortex – cytoplasmic alteration; ovary – cytoplasmic alteration; or eye – inflammation, chronic. The QAP confirmed all diagnoses of eye, cornea – inflammation, chronic. Moreover, the lungs of a few female mice were reviewed for the specific diagnoses of lung – hyperplasia, lymphoid and lung – inflammation, chronic. The QAP confirmed lymphoid hyperplasia in the lung of a single mouse in the 62.5 mg/kg dose group;

however, it was not verified in a single mouse of the 250 mg/kg dose group. Lung – inflammation, chronic was verified in a single mouse in the 62.5 dose group but not in two untreated controls. Furthermore, the salivary glands of many female mice were reviewed for the specific diagnoses of salivary gland – infiltration cellular and salivary gland – infiltration cellular, mononuclear cell. The QAP determined these changes to be common background findings and recorded them as “not remarkable”. The diagnosis of skin – hemangioma in a single female of the 250 mg/kg dose group was modified by the

QAP to skin, subcutaneous tissue – hemangioma by the QAP. This is preferred NTP nomenclature. A single diagnosis of stomach, glandular – hyperplasia, lymphoid was verified in an untreated control female.

AUDIT OF PATHOLOGY SPECIMENS

An audit of the remaining wet tissues for potentially untrimmed gross lesions indicated that all appeared to have been appropriately trimmed.

PHOTOGRAPHY

Photographs of representative lesions were taken and are listed in the photography log included with this report.

SUMMARY

The oral (gavage) administration of to male B6C3F1 mice at doses of 0, 62.5, 125, and 250 mg/kg resulted in increases in the incidences (or severity grades) in treated animals for the following diagnoses: stomach, glandular – inflammation, chronic;

stomach, glandular – pigmentation; stomach, glandular, epithelium – hyperplasia;

nose – inflammation; nose – polyp, inflammatory; nose, nerve – atrophy; nose, olfactory epithelium – atrophy; nose, olfactory epithelium – necrosis; nose, olfactory epithelium – respiratory metaplasia; nose, olfactory epithelium – degeneration; nose, respiratory epithelium – accumulation, hyaline droplet; nose, respiratory epithelium – hyperplasia; nose, respiratory epithelium – necrosis; liver – basophilic focus; liver – clear cell focus; liver – eosinophilic focus; liver – hepatoblastoma (combined incidence); liver – hepatocellular adenoma (combined incidence); and liver – hepatocellular carcinoma (combined incidence).

The oral (gavage) administration of to female B6C3F1 mice at doses of 0, 62.5, 125, and 250 mg/kg resulted in increases in the incidences (or severity grades) in treated animals for the following diagnoses: stomach, glandular – inflammation, chronic; stomach, glandular – pigmentation; stomach, glandular, epithelium – hyperplasia; nose – inflammation; nose, nerve – atrophy; nose, olfactory epithelium – atrophy; nose, olfactory epithelium – respiratory metaplasia; nose, olfactory epithelium – degeneration; nose, olfactory epithelium – accumulation, hyaline droplet; nose, respiratory epithelium – accumulation, hyaline droplet; nose, respiratory epithelium – hyperplasia; liver – eosinophilic focus; liver – hepatoblastoma (combined incidence); and liver – hepatocellular adenoma (combined incidence).

Neither proliferative lesions of the thyroid follicle (adenoma, carcinoma) nor random incidences of intestinal lymphangiectasia were related to test article exposure.

The decreased incidence of ovary – cyst was not considered to be related to test article exposure.

CONFLICT OF INTEREST STATEMENT

This statement is to certify that the personnel listed below, employed by

, participated in the Chronic Gavage

Toxicity/Carcinogenicity Study of in Male and Female B6C3F1 Mice at . They have not been involved in any aspect of the study for any organization other than NTP which conducted the study nor the generation and/or evaluation of materials or data which were reviewed prior to the receipt of materials from the study lab. Hence, their participation in the review poses no apparent or actual conflict of interest.

ETA…