Appendix_7__Sample_PWG_report.pdf

PATHOLOGY WORKING GROUP

FINAL REPORT

CHRONIC GAVAGE TOXICITY AND CARCINOGENICITY STUDY

OF IN B6C3F1 MICE )

Prepared by:

Pathology Working Group Coordinator

Research Triangle Park, NC 27709

Submitted to:

National Toxicology Program (NTP)/NIEHS Research Triangle Park, NC

The pathologist performing this review, , has had no involvement with any laboratory or organization concerned with this study other than the NTP and has not been involved in the origination or any previous review of data from this study.

PATHOLOGY WORKING GROUP

FINAL REPORT

Chronic Gavage Toxicity and Carcinogenicity Study of in B6C3F1 Mice

I. INTRODUCTION

A pathology working group (PWG) was convened on at the National Institute of Environmental Health Sciences (NIEHS), Research Triangle Park (RTP), NC. The participants were was an observer.

This PWG was conducted to histologically evaluate selected tissues from B6C3F1 mice administered via gavage for up to 105 weeks. This study was performed at

II. SUMMARY OF ORIGINAL STUDY RESULTS

a. Study Design

In this study, was administered to male and female B6C3F1 mice via gavage for 105 weeks at dosages of 0, 62.5, 125 or 250 mg/kg or a vehicle control (corn oil) for 5 days a week.

The dosages and number of animals examined microscopically per group are summarized below:

Dose (mg/kg) Males Females 0 (vehicle control) 50 50

62.5 50 50 125 50 50 250 50 50

b. Morbidity, Mortality, Body Weight and Clinical Observations

As stated in the study laboratory report1: the percent survival for males: control group=52; 62.5 mg/kg dosage group=62; 125 mg/kg dosage group and 250 mg/kg dosage group=65. The percent survival for females: control group=67; 62.5 mg/kg dosage group=80; 125 mg/kg dosage group=50; and the 250 mg/kg dosage group= 90* (*statistically significant at p≤ 0.05). After 105 weeks, body weights for all dosage levels for both males and females were within ±10 percent of the body weight of respective control animals. There were no related clinical observations reported.

c. Potential treatment-related effects

Administration of in B6C3F1 mice for 105 weeks was potentially associated with treatment-related increases in:

1. Liver

A. Hepatocellular carcinoma, multiple/hepatocellular, total in males B. Hepatocellular adenoma/hepatocellular adenoma, total in females C. Hepatocellular adenoma, multiple/hepatocellular, total in males D. Hepatoblastoma/hepatoblastoma, multiple/hepatoblastoma, total in males E. Eosinophilic foci in males and females F. Clear cell foci in males G. Mixed cell foci in males

2. Nose

A. Nerve, atrophy in males and females B. Glands, respiratory epithelium in males and females C. Olfactory epithelium degeneration in males and females D. Inflammation in females

3. Glandular Stomach

A. Epithelium, hyperplasia in males and females B. Epithelium, necrosis in males and females C. Inflammation, chronic and males and females

III. FINDINGS OF THE PWG

a. CONDUCT OF THE PWG

Before the PWG, the PWG Coordinator reviewed the study laboratory reports, including the SP’s pathology narrative, the summary and individual animal pathology tables, the pathology Quality Assessment (QA) reports, including the QA narrative with accompanying images, Pathology Data Review (PDR), and the tissues microscopically reviewed by the QAP. On

, a pre-PWG meeting was held at between the PWG Coordinator and NTP Pathologists to discuss the format of the PWG, resolve topography/terminology issues, resolve discrepancies between the SP and QAP in the diagnosis of several neoplastic to non-neoplastic and non-neoplastic to neoplastic lesions as listed in the QA narrative, and review examples of treatment-related and non-treatment-related lesions. The consensus diagnoses from slides reviewed by the NTP Pathologists and PWG Coordinator were documented on the PWG Coordinator’s slide review worksheets (SRW) attached to the end of this report.

The PWG Coordinator selected a set of slides for review by the PWG which included examples of potential treatment-related lesions, slides for which there were differences of opinion in diagnosis among the SP, QAP and/or the PWG Coordinator, as well as lesions added by the QAP or PWG Coordinator. There were nine voting PWG participants. The PWG Coordinator’s diagnosis was considered final in any cases in which there was not a consensus vote. The PWG consensus opinion for each slide, including any additional diagnoses made by the PWG, was recorded on the PWG Coordinator’s SRW attached to the end of this report.

Based on the study laboratory report and findings observed by the QAP and PWG Coordinator, the PWG was provided with the background that administration of to B6C3F1 mice for 105 weeks appeared to be associated with higher incidences of liver neoplasms as well as non-neoplastic lesions in the liver, nose, and glandular stomach of males and females when compared to the concurrent control groups. The potential target organs for this study were liver, nose, glandular stomach, thyroid (due to lesions observed from chemicals related to in male and female mice and ovary in female mice (due to decreased incidences of cysts in treated animals when compared to the concurrent control females).

b. RESULTS OF THE PWG REVIEW

Liver

The liver was considered a potential target organ because of increased incidences of neoplastic and non-neoplastic lesions in treated male and female mice when compared to concurrent controls. In general, the QAP and PWG Coordinator agreed with many of the SP’s neoplastic and non-neoplastic diagnoses. However, there were several discrepancies between the SP and QAP in the diagnosis of hepatocellular adenoma versus hepatocellular carcinoma; hepatocellular adenoma versus eosinophilic focus; and mixed cell focus versus other hepatocellular foci in both males and females. Examples of these discrepancies were reviewed by the PWG Coordinator and NTP Pathologists at the pre-PWG meeting and by the PWG panel.

Neoplastic Lesions

Hepatocellular carcinoma and hepatocellular carcinoma, multiple generally consisted of locally infiltrative, compressive, irregularly-shaped masses that replaced the normal hepatic parenchyma (Figure 1, Panel A). Neoplastic cells were often pleomorphic and arranged in thick trabeculae consisting of three or more cells. Cellular atypia and mitotic figures were frequently observed (Figure 1, Panel B). When there was more than one hepatocellular carcinoma present, it was recorded as “multiple”.

A. B.

Figure 1. Panel A - A hepatocellular carcinoma from LM 54 at 2X magnification. Note the irregular mass compressing the adjacent hepatic parenchyma (red arrow). Panel B - A higher magnification (20X) of Panel A. Note the neoplastic cells forming thickened trabeculae (black arrows) and the numerous mitotic figures (blue arrows).

Hepatocellular adenomas were relatively well-circumscribed masses that compressed the adjacent parenchyma (Figure 2, Panel A). Neoplastic cells were composed of well-differentiated hepatocytes that formed irregular hepatic plates of usually one to three cell layers thick and often compressed sinusoids (Figure 2, Panel B). In general, there was loss of normal lobular architecture with absence of central veins and portal tracts. The cytoplasm of the neoplastic cells varied tinctorially and often appeared eosinophilic, basophilic, clear, or mixed. When there was more than one hepatocellular adenoma observed, it was recorded as “multiple”.

Figure 2. Panel A - A hepatocellular adenoma from HF 397 at 2X magnification. Note the well-circumscribed mass compressing the adjacent hepatic parenchyma (red arrow). Panel B - A higher magnification (20X) of Panel A. Note the basophilic neoplastic cells forming irregular plates (black arrows) that compress sinusoids and the absence of central veins and portal triads.

Hepatoblastomas were usually well-demarcated, irregularly-shaped masses (Figure 3, Panel A) occasionally found within hepatocellular carcinomas or adjacent to hepatocellular adenomas or hepatocellular carcinomas. The neoplastic cells were small with hyperchromatic nuclei and arranged in sheets (Figure 3, Panel B) or occasionally in rows surrounding vascular spaces.

When there was more than one hepatoblastoma observed, it was recorded as “multiple”. When a hepatoblastoma arose within a hepatocellular carcinoma, only the hepatoblastoma was diagnosed. In general, there was good agreement between the SP, QAP, and PWG Coordinator in diagnosing hepatoblastomas.

Hepatocholangiocarcinomas were expansile masses (Figure 3, Panel C) composed of malignant-appearing hepatocytes and bile duct epithelial cells (Figure 3, Panel D). The hepatocellular component usually consisted of neoplastic cells forming thick trabeculae or sheets and the biliary component consisted of neoplastic cells forming small nests with a mild to moderate amount of connective tissue stroma.

C. D.

Figure 3. Panel A - A hepatoblastoma from HM 163 at 2X magnification. Note the well-demarcated mass compressing the adjacent hepatic parenchyma (red arrow). Panel B - A higher magnification (20X) of Panel A. Note the small, basophilic neoplastic cells (black arrow). Panel C - A hepatocholangiocarcinoma from LM 53 at 2X magnification. Note the expansile mass compressing the adjacent hepatic parenchyma (yellow arrow). Panel D - A higher magnification (20X) of Panel C. Note the neoplastic hepatocytes (black arrows) and neoplastic bile duct epithelial cells (blue arrows).

In addition, there was good agreement between the SP, QAP, and PWG Coordinator in the diagnosis of several neoplasms in the liver of control and treated males and females. These tumors were infrequent and included hemangioma, hemangiosarcoma and hemangiosarcoma, multiple.

Non-neoplastic Lesions

Eosinophilic foci were observed more frequently in treated males and females when compared to the concurrent control groups. Clear cell and mixed cell foci were present more often in treated males when compared to control males. Eosinophilic foci were characterized by focal regions of enlarged hepatocytes with granular, pink cytoplasm (Figure 4, Panel A). Clear cell foci consisted of a focus of hepatocytes that were of normal size to slightly enlarged containing cytoplasmic clear space and centrally located nuclei (Figure 4, Panel B). Mixed cell foci consisted primarily of hepatocytes with increased cytoplasmic basophilia and clear cytoplasm (Figure 4, Panel C) or cytoplasmic eosinophilia and clear cytoplasm. In these instances neither of the cell types exceeded 80% of the focus. Large eosinophilic, clear cell, or mixed cell foci occasionally compressed the adjacent hepatic parenchyma.

Fatty change, characterized by the presence of variable-sized clear vacuoles within randomly scattered hepatocytes, was observed in control and treated males and females. The incidences and severity of this lesion were similar in treated and control animals (Figure 4, Panel D). There was good agreement in the diagnosis of fatty change by the SP, QAP, and PWG Coordinator.

C. D.

Figure 4. Panel A - An eosinophilic focus from HF 382 at 4X magnification (black arrow). Panel B - A clear cell focus from HM 157 at 4X magnification (red arrow). Panel C - A mixed cell focus from MF 320 at 4X magnification (blue arrow). Panel D – Fatty change within hepatocytes from MF 306 at 4X magnification (yellow arrows).

Nose

There were diagnostic criteria differences between the SP and QAP concerning lesions involving olfactory and respiratory epithelium. In general, the SP used the term “olfactory epithelium degeneration” to collectively describe the following lesions used by the QAP: “olfactory epithelium degeneration”, “olfactory epithelium, respiratory metaplasia”, and “olfactory epithelium atrophy”. The QAP used the term “respiratory epithelium hyperplasia” to describe lesions involving strictly the respiratory epithelium as well as lesions associated with submucosal glands lined by respiratory epithelium.

There were several potential treatment-related nasal lesions in males and females identified by the SP and/or QAP that the PWG Coordinator agreed were present. These lesions included nerve atrophy; olfactory epithelium respiratory metaplasia; olfactory epithelium atrophy; olfactory epithelium degeneration; respiratory epithelium hyperplasia; and inflammation. Examples of these lesions were reviewed by the PWG Coordinator and NTP Pathologists at the pre-PWG meeting and by the PWG panel.

Nerve atrophy was a prominent lesion in many of the treated animals when compared to concurrent controls (Figure 5, Panel A), with females being affected more often than males. This lesion consisted primarily of nerve bundles that were smaller in diameter and fewer in number in the lamina propria (Figure 5, Panel B) of the olfactory region in predominantly Level III (dorsal aspect, septum, and ethmoturbinates) and occasionally in Level II (dorsal meatus). In general, the overlying epithelium associated with nerve atrophy was metaplastic, appearing as respiratory epithelium. In addition, Bowman’s glands were frequently mildly to markedly dilated and replaced nerve bundles. In general, there was good agreement between the SP and the QAP in diagnosing this lesion. The PWG Coordinator added a few more diagnoses, including several to the control female group. The PWG confirmed this lesion and accepted the request by the NTP pathologists to add the site modifier “olfactory epithelium” to the diagnosis of nerve atrophy.

Olfactory epithelium respiratory metaplasia was observed most frequently along the dorsal region, nasal septum, and ethmoturbinates in Level III and occasionally the dorsal meatus of Level II, in predominantly treated animals but occasionally in controls. The incidences and severity of this lesion increased with increasing dosage. Olfactory epithelium respiratory metaplasia consisted of minimal to moderate replacement of olfactory epithelium by ciliated, columnar/cuboidal, respiratory epithelial cells (Figure 5, Panel B). This lesion was often associated with olfactory epithelium nerve atrophy. The QAP added the term olfactory epithelium respiratory metaplasia; however, the SP described the finding as a part of olfactory degeneration. The PWG confirmed the finding of olfactory epithelium respiratory metaplasia.

Figure 5. Panel A - Olfactory epithelium (red arrow) and nerve bundles (black arrow) from control animal, VF 225, at 20X magnification. Panel B - Olfactory epithelium nerve atrophy from HF 372 at 20X magnification. Note the decrease in number and size of nerve bundles (blue arrows) with mildly to markedly dilated Bowman’s glands replacing them (purple arrows) and respiratory metaplasia of the olfactory epithelium (green arrow).

Olfactory epithelium atrophy was a diagnosis added by the QAP and observed at a higher frequency in treated males and females, and rarely in control animals. Olfactory epithelium atrophy was described by the SP as olfactory epithelium degeneration. This was predominantly a lesion of minimal to moderate severity and present along the dorsal region, septum, and ethmoturbinates of Level III and very seldom involving the dorsal meatus of Level II. Olfactory epithelium atrophy was characterized by focal to diffuse thinning of the epithelium (Figure 6, Panel B) when compared to the epithelium from concurrent control animals (Figure 6, Panel A).

The PWG confirmed this finding.

Figure 6. Panel A - Normal olfactory epithelium (red arrow) from control animal, VF 225, at 40X magnification.

Panel B - Olfactory epithelium atrophy from MF 324 at 40X magnification. Note the thinning of the epithelium (blue arrow) when compared to the epithelium in Panel A.

Olfactory epithelium degeneration was observed most often in treated males and females at the dorsal region and/or ethmoturbinates of Level III and occasionally at the dorsal meatus of Level II. This lesion was characterized by hypocellularity and disorganization (often with increased clear intercellular space) of olfactory epithelial cells that were often swollen (Figure 7, Panel B) with or without an overall decrease in height of the epithelium when compared to the epithelium from concurrent control animals (Figure 7, Panel A). This lesion was observed by the SP, QAP, and PWG Coordinator. The QAP and PWG Coordinator diagnosed this finding less often than the SP due to the “lumping” of several lesions under the term “degeneration” by the SP. The PWG confirmed this diagnosis.

Figure 7. Panel A - Normal olfactory epithelium (black arrow) from control animal, VF 225, at 40X magnification.

Panel B - Olfactory epithelium degeneration from HF 372 at 40X magnification. Note the hypocellularity, increased clear intercellular space and disorganization of swollen cells in the epithelium (blue arrow).

The QAP added the diagnosis of olfactory epithelium hyaline droplet accumulation and respiratory epithelium hyaline droplet accumulation in control and treated animals of both sexes.

Olfactory epithelium hyaline droplet accumulation was observed most often in the ventral ethmoturbinates in Level III but also noted in the dorsal portions of Level II and Level III.

Hyaline droplet accumulation consisted of intracytoplasmic, homogenous, eosinophilic, globular material in olfactory epithelial cells that often displaced nuclei (Figure 8, Panel A). The severity of this lesion was increased in high dose females. Respiratory epithelium hyaline droplet accumulation was primarily observed in the ventral respiratory epithelium (adjacent to the squamous-respiratory junction in Level II and to the olfactory-respiratory junction in Level III).

Hyaline droplet accumulation consisted of intracytoplasmic, homogenous, eosinophilic, globular material in respiratory epithelial cells that at times displaced nuclei (Figure 8, Panel B). The severity of this lesion was increased in high dose females. The PWG panel confirmed the presence of hyaline droplets in both the olfactory and respiratory epithelium.

Figure 8. Panel A – Hyaline droplet accumulation in olfactory epithelial cells of ethmoid turbinates in Level III (black arrow) of HF 380 at 20X magnification. Panel B - Hyaline droplet accumulation in respiratory epithelial cells along the septum at Level II (blue arrow) of HF 380 at 20X magnification.

The QAP added the diagnoses of respiratory epithelium hyperplasia in control and treated animals of both sexes. The severity of respiratory epithelium hyperplasia was increased in high dose males and females when compared to concurrent controls and other dosage groups. As stated in the QA report2, this finding “was typically observed at Level II and less frequently at Level I and was characterized by proliferation of ciliated columnar cells that occasionally formed villous structures that projected into nasal passages” (Figure 9, Panel A). “Frequently, these cells extended into the glands of the subjacent lamina propria that were often dilated and hyperplastic” (Figure 9, Panel B). This finding was observed by the SP but diagnosed as “nose, glands, respiratory epithelium-hyperplasia” In an effort to sustain consistency with the NTP nomenclature, the QAP removed the site, “glands” from the diagnosis. The PWG Coordinator and PWG panel were in agreement with this terminology change and the lesion was confirmed at the PWG.

Increased incidences of inflammation of minimal to mild severity were observed in high dose females when compared to concurrent controls and males. This lesion consisted of accumulations of neutrophils, homogenous eosinophilic material (proteinaceous fluid), and a few mononuclear cells (Figure 9, Panel B) in all levels of the nose. Sometimes this inflammation was associated with the presence of plant/food-like material or hair shafts (predominantly in the ventral portions of Levels I and II). In these instances, the PWG Coordinator added the diagnosis of “foreign body” which per the NTP Pathologists, was to be accepted.

Figure 9. Panel A - Respiratory epithelium hyperplasia in Level I of LM 60 at 10X magnification. Note villous structures formed by hyperplastic epithelium along the dorsal meatus (black arrow). Panel B - Respiratory epithelium hyperplasia in Level II of control, VM 2, at 20X magnification. Note increased numbers of dilated glands lined by hyperplastic respiratory epithelial cells (blue arrows). Also note minimal inflammation within the nasal passage (red arrow).

Glandular Stomach

The glandular stomach was considered a potential target organ because of increased incidences of epithelium hyperplasia, epithelium necrosis, and chronic inflammation in treated male and female mice when compared to concurrent controls. In general, the QAP and PWG Coordinator agreed with many of the SP’s diagnoses of epithelium hyperplasia and chronic inflammation.

However, the QAP did not agree that necrosis was present and added the diagnosis of pigment.

The PWG Coordinator concurred with the QAP. Examples of these lesions were reviewed by the PWG Coordinator and NTP Pathologists at the pre-PWG meeting and by the PWG panel.

Hyperplasia of the glandular stomach epithelium was characterized by the increased presence of minimal to mild numbers of parietal cells, and occasionally chief cells, when compared to controls (Figure 10, Panels A and B). Often accompanying hyperplasia was minimal to mild inflammation. Chronic inflammation consisted of a slight increase in the numbers of randomly scattered mononuclear and fewer polymorphonuclear cells subjacent to the muscularis mucosae.

Numerous macrophages, some containing intracytoplasmic golden brown pigment, were also observed in these areas (Figure 10, Panel B). The SP described the criteria for necrosis as “degenerative cytologic changes in or absence of epithelium, dilatation of gastric pits, or pigment in the lumen of gastric pits”.1 The QAP did not observe notable gastric epithelial degeneration or regard the gastric pit dilatation and pigment accumulation as necrosis and opted to diagnose the pigment accumulation only. Pigmentation was characterized by the presence of intracytoplasmic golden brown pigment (presumptive test material) in macrophages as well as similar golden brown within gastric pits (Figure 10, Panel B). The PWG panel confirmed the presence of glandular stomach epithelium hyperplasia, chronic inflammation, and pigmentation but did not confirm necrosis.

Figure 10. Panel A - Glandular stomach epithelium from control animal, VM 6, at 20X magnification. Panel B – Glandular stomach epithelium hyperplasia, inflammation, and pigmentation in HF 385 at 20X magnification. Note increased numbers of parietal cells (blue arrows) when compared to the control animal in Panel A. Also note minimal chronic inflammation (red arrows) and golden brown pigment within gastric pits (black arrows).

Miscellaneous Findings

The NTP Pathologists and PWG Coordinator and/or the PWG panel reviewed examples of several lesions that were considered to most likely be unrelated to administration of The following was concluded:

• The severity grade of ovary-cyst will be removed.

• All diagnoses of tooth-dysplasia made by the QAP in female mice will be accepted.

IV. HISTOTECHNIQUE QUALITY

The overall histological processing and slide preparation was good. There were no artifacts that interfered with the interpretation of tissue sections.

V. POST-PWG ACTION ITEMS

There were no post-PWG action items.

VI. CONCLUSIONS OF THE PWG (Summary)

The results of this PWG review generally confirmed the SP’s finding of increased neoplastic and non-neoplastic lesions in the liver of treated males and females when compared to controls. This review also confirmed the diagnoses of non-neoplastic lesions in the nose and stomach of male and female mice. administration primarily targeted the olfactory epithelium in the nasal cavity potentially causing non-neoplastic lesions such as nerve atrophy, epithelium atrophy, respiratory metaplasia, and degeneration in males and females. Common lesions observed in the glandular stomach of animals treated with included epithelium hyperplasia, inflammation, and pigmentation. The PWG panel confirmed the presence of most of these lesions that were considered potentially treatment-related.

administration for 105 weeks in mice did not affect the thyroid gland, as lesions reported following treatment with chemicals similar to were not observed in this study. The ovary was initially considered a potential target organ due to decreases in incidences of cysts. The PWG panel reviewed examples of these cysts and commented that ovarian cysts were common in older mice and most likely not due to administration. In addition, the PWG did not confirm the presence of lymphangiectasis in the small intestines of four male mice in which the QAP recorded the finding. Lymphatic ectasia was diagnosed in the duodenum, jejunum, and mesenteric lymph nodes of rats administered via gavage for 3 months3 so it was requested that these tissues be evaluated for similar lesions in the chronic mouse study.

PRE‐PWG WORKSHEETS

Chemical Number:

Date: Dot Color: Blue

Dot No.

Animal Number

# of Slides

Organ Diagnosis Consensus

1 46 1 Harderian gland ‐ 09 Infiltrate Cell, Mononuclear, Minimal or No Remarkable Lesion

No Remarkable Lesion

2 353 1 Harderian gland ‐ 09 Atrophy, Minimal or Dilatation, Minimal

Dilatation, Minimal

3 111 1 Harderian gland ‐ 09 Adenoma ‐ Yes or No Adenoma

4 313 1 Harderian gland ‐ 09 Hyperplasia, Moderate or Adenoma

Adenoma

5 81 1 Harderian gland ‐ 09 Adenoma or Carcinoma Carcinoma

6 346 1 Harderian gland ‐ 09 Carcinoma or Hyperplasia, Moderate

Carcinoma

Sex and Species: Male and Female Mice

Chemical Name:

PRE‐PATHOLOGY WORKING GROUP DIAGNOSIS

7 17 1 Nose, Olfactory Epithelium ‐ 01 Degeneration, Minimal ‐ Yes or No Degeneration, Minimal

Nose, Olfactory Epithelium ‐ 01 Atrophy, Moderate ‐ Yes or No Atrophy, Moderate

8 200 1 Nose, Olfactory Epithelium ‐ 01 Degeneration, Mild ‐ Yes or No Degeneration, Mild

Nose, Olfactory Epithelium ‐ 01 Respiratory Metaplasia, Minimal ‐ Yes or No

Respiratory Metaplasia, Minimal

9 120 1 Nose, Olfactory Epithelium ‐ 01 Degeneration, Moderate ‐ Yes or No

Degeneration, Moderate

Nose, Olfactory Epithelium ‐ 01 Respiratory Metaplasia, Moderate ‐ Yes or No

Respiratory Metaplasia, Moderate

Nose, Nerve ‐ 01 Atrophy, Minimal ‐ Yes or No Atrophy, Minimal

10 372 1 Nose, Olfactory Epithelium ‐ 01 Degeneration, Moderate ‐ Yes or No

Degeneration, Moderate

Nose, Olfactory Epithelium ‐ 01 Respiratory Metaplasia, Moderate ‐ Yes or No

Respiratory Metaplasia, Moderate

Date: Dot Color: Blue

Dot No.

Animal Number

# of Slides

Organ Diagnosis Consensus

Sex and Species: Male and Female Mice

Chemical Name:

PRE‐PATHOLOGY WORKING GROUP DIAGNOSIS

11 17 1 Liver ‐ 12 Hepatocellular Adenoma ‐ Yes or No

Hepatocellular Adenoma

12 143 2 Liver ‐ 10, 11 Hepatocellular Adenoma or Hepatocellular Carcinoma

Hepatocellular Adenoma on slide #11; Hepatocellular Carcinoma on slide #10 13 387 1 Stomach Gland, Epithelium ‐ 05 Necrosis, Minimal ‐ Yes or No Necrosis Not Present 14 388 1 Stomach Gland, Epithelium ‐ 05 Necrosis, Minimal ‐ Yes or No Necrosis Not Present

15 280 1 Pituitary Gland ‐ 06 Pars Distalis ‐ Adenoma or Hyperplasia, Moderate

Pars Distalis ‐ Adenoma

16 294 1 Heart ‐ 03 Inflammation, Chronic, Moderate ‐ Yes or No

Inflammation, Chronic, Moderate

Cardiomyopathy, Mild ‐ Yes or No Cardiomyopathy, Mild

17 48 1 Kidney ‐ 04 Mineralization, Minimal or No Remarkable Lesion

No Remarkable Lesion

Animal No.

Tissue SP's Diagnosis RP's Diagnosis Consensus

18 Liver Hepatoclr Aden Eosin Focus Take to PWG 20 Liver Hepatoclr Aden Eosin Focus Hepatoclr Aden 25 Stom Forestom Sq Cl Papillom Epithelium ‐ Hyperplasia Epithelium ‐ Hyperplasia 26 Liver Hepatoclr Aden Mixed Cl Focus Mixed Cl Focus

Thyroid Gl ‐ Follicular Cell

Adenoma Follicle ‐ Degen Take to PWG

34 Liver Hepatoclr Aden Mixed Cl Focus Hepatoclr Aden

34 Thymus Thymoma Bgn Epithelial Cell ‐ Hyperplasia Thymoma Bgn

59 Liver Hepatoclr Aden Eosin Focus Hepatoclr Aden 63 Liver Hepatoclr Aden Basoph Focus Take to PWG

Lymph Node, Pancreatic

Lymph Mal Necrosis Necrosis

69 Stom Forestom Sq Cl Papillom Epithelium ‐ Hyperplasia Epithelium ‐ Hyperplasia 70 Liver Hepatoclr Aden Eosin Focus Eosin Focus

71 Bone Marrow Histio Sarc Autolysis Precludes

Examination Histio Sarc

71 Liver Histio Sarc Autolysis Precludes

Examination Histio Sarc

Skin, Sebaceous

Gl Carcinoma

Harderian Gl ‐ Inflammation, Chronic

Harderian Gl ‐ Inflammation, Chronic

99 Liver Hepatoclr Aden Eosin Focus Eosin Focus

PRE‐PWG MEETING ON

99 Liver Hepatoclr Aden Eosin Focus Eosin Focus 103 Liver Hepatoclr Aden Eosin Focus Hepatoclr Aden

104 Skin, Epidermis Keratoacanthoma Insufficient Tissue Keratoacanthoma

123 Liver Hepatoclr Aden Eosin Focus Hepatoclr Aden

130 Spleen Hemangiosarcoma Autolysis Precludes

Examination Hemangiosarcoma

Thyroid Gl ‐ Follicular Cell

Carcinoma Thyroid Gl ‐ Inflammation, Chronic Thyroid Gl ‐ Inflammation, Chronic 184 Epididymis Hemangioma Angiectasis Angiectasis

Liver Hepatoclr Aden, Multiple Eosin Focus Hepatoclr Aden, Multiple

200 Liver Hepatoclr Aden, Multiple Eosin Focus Take to PWG

Thyroid Gl ‐ Follicular Cell

Adenoma Not Present in Section Not Present in Section

Pituitary Gl, Pars

Distalis Adenoma Hyperplasia Hyperplasia

212 Ovary Cystadenoma Cyst Take to PWG 217 Spleen Hemangiosarcoma Angiectasis Angiectasis

Pituitary Gl, Pars

Intermed Adenoma Hyperplasia Adenoma

Tissue SP's Diagnosis RP's Diagnosis Consensus

PRE‐PWG MEETING ON

Pituitary Gl, Pars

Distalis Adenoma Insufficient Tissue Insufficient Tissue

231 Ovary Cystadenoma Not Present in Section Take to PWG

242 Adrenal Medulla Pheochromocytoma, Bgn Hyperplasia Hyperplasia

249 Int Lg Cecum Lymph Mal Lymphoid Tissue ‐

Hyperplasia Lymph Mal

Int Sm Ileum Lymph Mal Lymphoid Tissue ‐

Hyperplasia Lymph Mal

269 Lung Hepatoclr Carc, Metastatic

Not Present in Section Hepatoclr Carc, Metastatic

Kidney, Renal

Tubule Adenoma No Section Present No Section Present

Pituitary Gl, Pars

Distalis Adenoma Hyperplasia Hyperplasia

309 Adrenal medulla Pheochromocytoma, Bgn Hyperplasia Hyperplasia

Parathyroid

Gland Adenoma Not Present in Section Not Present in Section

346 Harderian Gl Carcinoma Hyperplasia Carcinoma 353 Spleen Histio Sarc Hyperplasia, Mast Cell Hyperplasia, Mast cell

39 Liver ‐ Hepatoclr Aden, Multiple Hepatoclr Aden, Multiple

Kidney, Renal

Tubule ‐ Hepatoclr Carc, Metastatic Hepatoclr Carc, Metastatic

91 Liver ‐ Hepatoclr Aden, Multiple Hepatoclr Aden, Multiple

93 Liver ‐ Hepatoclr Aden, Multiple Hepatoclr Aden, Multiple

111 Harderian Gl ‐ Adenoma Adenoma 112 Harderian Gl Hyperplasia Adenoma Hyperplasia 131 Liver ‐ Hepatoclr Aden Hepatoclr Aden 136 Stom Gland Normal Carcinoma, Metastatic Carcinoma, Metastatic 137 Liver ‐ Hepatoclr Aden Hepatoclr Aden

140 Lung (Hyperplasia dx'd in Rpt

14) Alv Bron Aden Hyperplasia

141 Liver Mixed Cl Focus Hepatoclr Aden, Multiple Hepatoclr Aden, Multiple

Testes, Interstit

Cel ‐ Adenoma Adenoma

155 Stom Gland ‐ Carcinoma, Metastatic Carcinoma, Metastatic 158 Liver ‐ Hemangiosarcoma Hemangiosarcoma

Islets Panc Hyperplasia Adenoma Adenoma

Tissue SP's Diagnosis RP's Diagnosis Consensus

PRE‐PWG MEETING ON

Adrenal Cortex, Subcapsular

‐ Adenoma Adrenal Cortex ‐

Hyperplasia, Mild (6)

Liver Mixed Cl Focus Hepatoclr Aden Take to PWG 215 Liver Mixed Cl Focus Hepatoclr Aden Mixed Cl Focus

Int Sm

Duodenum Normal Adenoma Normal

236 Int Sm Jejunum ‐ Leiomyosarcoma Leimyosarcoma

Adrenal Cortex, Subcapsular

‐ Adenoma Adenoma

Liver Basoph Focus Hepatoclr aden Take to PWG

Ovary, Corpus

Luteum Hyperplasia Ovary ‐ Granlsa Tm Bgn

Ovary ‐ Inflammation, Chronic, Moderate (7)

250 Adrenal Cortex ‐ Adenoma No Remarkable Lesion (6)

258 Liver Mixed Cl Focus Hepatoclr Aden Take to PWG 265 Liver ‐ Lymph Mal Lymph Mal

Kidney Normal Lymph Mal Lymph Mal Spleen Hema Cell Prol Lymph Mal Lymph Mal

266 Liver Lymph Mal No Remarkable Lesion266 Liver ‐ Lymph Mal No Remarkable Lesion Lung ‐ Lymph Mal No Remarkable Lesion

Lym Node Mesen ‐ Lymph Mal No Remarkable Lesion

Kidney Normal Lymph Mal Normal Spleen Normal Lymph Mal Normal

277 Harderian Gl ‐ Adenoma Adenoma Liver ‐ Hepatoclr Aden Eosin Focus (3)

279 Spleen Hema Cell Prol Lymph Mal Lymphoid Hyperplasia, Moderate (4)

Lymph Node

Mesen Normal Lymph Mal

Lymph Mal and add Spleen ‐ Lymph Mal (4)

313 Harderian Gl Hyperplasia Adenoma See Blue Dot Pre‐PWG group 342 Nose ‐ Lymph Mal No Remarkable Lesion

Bone Normal Lymph Mal Normal

Stom Gland Inflammation, Chronic Lymph Mal Inflammation, Chronic

351 Uterus ‐ Polyp Stromal Polyp Stromal 352 Uterus ‐ Polyp Stromal Polyp Stromal 353 Uterus ‐ Polyp Stromal Polyp Stromal 397 Uterus ‐ Polyp Stromal Polyp Stromal

Tissue SP's Diagnosis RP's Diagnosis Consensus

PRE‐PWG MEETING ON

399 Ovary Angiectasis Hemangiosarcoma Angiectasis

Date: Dot Color: Green

Dot No.

Animal Number

# of Slides Organ Diagnosis Consensus

1 205 1 Tooth ‐ 01 Dysplasia, Minimal Agree 2 360 1 Tooth ‐ 01 Dysplasia, Minimal Agree 3 336 1 Tooth ‐ 01 Dysplasia, Minimal Agree 4 256 1 Tooth ‐ 01 Dysplasia, Minimal Agree 5 258 1 Tooth ‐ 01 Dysplasia, Minimal Agree

Sex and Species: Female Mice

Chemical Name:

PRE‐PATHOLOGY WORKING GROUP DIAGNOSIS

PWG COORDINATOR’S WORKSHEETS