Document zbMMLr6dnJ2GQ0V67J7ZaYVym

2.77 Pathology Associates International A Company of Science Applications temational Corporation ARGOS TL == S= e ar iaeComes PATHOLOGY REPORT (ANCILLARY STUDY) ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG AND LIGHT MICROSCOPIC EVALUATION OF LIVER IN CRL:CD@BR VAF/PLUS RATS) `ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS CLIENT STUDY NUMBER 418-013 PAI STUDY NUMBER 99-11 (EM 99.39) Prepared by: James B. Nold, D.V.M., Ph.D., Diplomate, A.C.V.P. Pathology Associates International 4915-D Prospectus Drive Durham, NC 27713 April 5,1999 PREPARED FOR 3M CORPORATE TOXICOLOGY, 3M CENTER, ST. PAUL, MN 55144-1000 0154). e 2915e D Prospectt us Drive +r Durham, No-- rth Carolin-- a 2e77e13e-- (919S ) 544-5-- 257 + (919)-- 5e 44-3218e FAX Pathology Narrative Light Microscopy and Transmission Electron Micrograph Interpretation Forms Transmission Electron Micrographs Quality Assurance Statement Section 1 o m v 001543 I Pathology Narrative 001544 PrAmneciilclaarAyrPgauoslNoogyS1R8ep0o1r3t PATHOLOGY REPORT (ANCILLARY STUDY) ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG AND LIGHT MICROSCOPIC EVALUATION OF LIVER IN CRL:CDBR VAF/PLUS RATS) ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS CLIENT STUDY NUMBER 418-013 PAI STUDY NUMBER 99-11 (EM 99.39) SUMMMARY Ultrastructural evaluation of liver and lungsofrat pups from dams treated with 0 (control), 0.4, 1.6, or 3.2 mg/kg/day (high-dose) PFOS identified increased numbers of peroxisomes within hepatocytes of high-dose rat pup livers. Quantitation of peroxisomes indicated at least twice the number of peroxisomes in animals treated with 0.4 mg/kg/day and triple the number in those given 1.6 or 3.2 mg/kg/day. Differences in cellular `membranes or mitochondria were not discernible between control and treated animals in the samples examined; however, suboptimal fixation precluded a detailed evaluation of these organelles. In lung samples, the presence of type II pneumocytes and lamellar bodies was evaluated. No significant ultrastructural differences were identified between lung from control and treated rats. Random type II pneumocytes containing lamellar bodies were identified in all lungs examined ultrastructurally. PROCEDURES The objective of this study was to examine by electron microscopy the livers and lungs from selected day-21 gestation rat fetuses in an oral (gavage) pharmacokinetic study of PFOS. In addition to ageneral qualitative evaluationfordifferences between control and treated animals, the ultrastructural endpoints included quantitation of peroxisomes within hepatocytes, membrane and mitochondrial changes in hepatocytes, and the quantitation of surfactant lamellar bodies in the lung. Light microscopy of paraffin-embedded liver sections was also performed. For the pharmacokinetic study, 90 female Crl:CDBR VAF/PLUS rats were given the test article or the vehicle once daily beginning 42 days prior to cohabitation with a male through day 20 postpartum according to Text Table 1. ol 001545 PriAmnediialArPgiuhscNioo.gy41R5e.p0o1s3 Growy | Female Ras ngigrann mga) nig) [C01 16 1oven[oT 5 1] Cu vw [or[oe 75) Co 6[0s ToTs Cov 6Te[om TT 5 7} Cv we T13 oe 2 1 5 1| At Caesarean-sectioning on day 21 of presumed gestation, lung and liver samples were collected from three fetuses per litter from five litters per dosage group. Samples of lung and liver from selected fetuses for Groups I, IIL, IV, and V were thin-sliced and placed in McDowell-Trump fixative (McDowell EM, 1976) and submitted to this laboratory for electron microscopy processing and evaluation. Sections of liver from these same animals were immersion-fixed in neutral buffered formalin and submitted along with electron microscopy samples. Per sponsor request, tissue samples from selected pups were processed and evaluated (pTuepxtnuTmabbleer 2f).roTmhtehatfidvaemd'igsitlintuermber is the dam number; the suffixes 1-4 denotes the eT Toi mTor || cool Gmpiatsn | osm195a361o L1o95m0a.1s 32 95m6a8rt2en | ess 195363 195602 19560-1 119355015612 191594564.612 115935662232 119955777943 = 818[8 | The lungs and livers were then processed into Spurr's resin for ultrastructural examination. Thick (Ij) sections were cut, stained with toluidine blue, and examined to select representative areas for further electron microscopy processing. Thin sections (approximately 90 nm) were cut, mounted on 200-mesh copper grids, stained with 5% `methanolic uranyl acetate and Reynolds lead citrate, and examined on a Zeiss 900 transmission electron microscope. Centrilobular hepatocytes, where clearly identifiable in liver sections, were preferentially examined. Representative electron photomicrographsofliver and lung were taken and significant ultrastructural features `were summarized for each photograph and animal on a designated transmission electron `micrograph interpretation form. The number of peroxisomes in hepatocytes was `manually counted for each center photographed hepatocyte and recorded. For light microscopy, formalin-fixedliversamples were processed and embedded in paraffin, cut at approximately five microns, stained with hematoxylin and eosin, and evaluated. 2 001545 PriAmnecdililcaarAyrPgautshoNloo.gy41R8e.p0o1r3t RESULTS AND DISCUSSION Individual interpretations of light microscopy slides and electron micrographs for each animal are shown in Section II. Selected representative electron photomicrographs were labeled and are shown in Section IIL Light Microscopy. A summary of the incidence and severity of light microscopic findings in the liver is shown in Text Table 3. There was an increased incidence and severity of eosinophilic granularity of the cytoplasm of hepatocytes in treated rats compared with control rat livers. The severityofthis finding increased in a dose-related fashion. Hepatocellular eosinophilic granularity was diffusely distributed in most animals given 1.6 or 3.2 mg/kg/day of PFOS, but was predominantly periportal in 1 animal given 1.6 and in 3 animals given 0.4 mg/kg/day. Cytoplasmic clearing of hepatocytes was prominent in control livers. This morphological description is usually associated with the normal accumulation ofglycogen in hepatocytes. In this study, the incidence and severity of cytoplasmic clearing was almost inversely proportional to the occurrence of eosinophilic granularity. `The liver is a hematopoietic organ in rat fetuses and neonates. Thus, large numbers of hematopoietic cells are present within the sinusoids throughout the liver. Extramedullary hematopoiesis (EMH) occurred in all animals, including controls, and was uniformly graded as moderate to moderately-severe. The severity of EMH was increased in the treated groups. Random mitoses of hepatocytes are normal in rat livers, especially in fetuses and neonates. There appeared to be a light increase in hepatocellular mitotic figures in the animals given 3.2 mg/kg/day PFOS. 0 04 6 32 mE p r increased misniigmhatl 23 31 moderate : i modesrlaitgeht ii sigh | - : 2 3 001549 PriAmnecdliacseyArPpatshoNloogyS1R3e0p1o3r [Electron Microscopy. `Fixation of the tissues was less than optimal for electron microscopy. This was especially evident in the liver compared with the lung. Lung is inherently less dense than liver and allows better infiltration of the fixative into the tissue. Suboptimal fixation was characterized by clumpingoflarge organelles, discontinuous plasma membranes, loss of detailofcytoplasmic organelles, dilatationof endoplasmic reticulum and the nuclear envelope, granular deposits in the mitochondrial matrix, and dilatation of mitochondrial cristae. The poor fixation of membranes made ultrastructural assessment of these structures impossible. Fixation, however, was still considered adequate to assess for the presence of peroxisomes in the hepatocytes and lamellar bodies in the lung and to make `general comparisons between control and treated samples. LIVER Control (0 mg/kg/day). Subcellular featuresofcontrol hepatocytes are demonstrated in Figures 1 and 2. Hepatocytes characteristically contain numerous mitochondria and prominent endoplasmic reticulum. Clumping of the large organelles (mitochondria, rough endoplasmic reticulum and peroxisomes) is indicative of poor fixation. In these cells rough endoplasmic reticulum was evident, but smooth endoplasmic reticulum was not because of suboptimal fixation. Mitochondria generally appeared dark and contained scattered dense deposits and some dilated cristae, both artifacts due to suboptimal fixation. Dilatationof endoplasmic reticulum and the nuclear envelope were additional artifacts of poor fixation. The fine granular material in the cytoplasm is consistent with glycogen particles. Peroxisomes were identified and quantitated in from representative hepatocytes. For the control livers, the average number of peroxisomes counted per hepatocyte photographed was 4.2 peroxisomes per cell (Text Table 4). Fetal and neonatal rtahte lliivveerr issinaushoeimdast,opoietic tissue, and numerous hematopoietic cells were evident within [sNuombesr| Per3on2iomeTs|wNoumbser]| pe1ra2i0smeTs|oNsumboer|]pe1on1is6omeTs|heNosmbeess|]Porcouosom|es [[Fioss0s5o:1[| 3a0s [Joisosess||_15202 T|osoeweo0rs||t10228 TTttoosseon-i1|| 1t3o0e || [[oossoio-e1a|]_ss2o TJiissssitee1r[]13si0 JTi1oossea2a3|] 2t0o0s TTiloossnmoaa|] a1i1s2 || Dnesee e2 r oTroTovemmsn=pr Fe1o07To TeMeepma=oeeys o1em3e0d 7|wVmeaw= 126] 0.4 mg/kg/day. The average numberofperoxisomes per hepatocyte (10.1) was higher in the animals given 0.4 mg/kg/day PFOS compared with controls (Text Table 4). A representative hepatocyte is shown in Figure 3. Qualitatively, there were no identifiable 4 00154% PriAmnecdiilclaarAyrPgautshoNloo.gy41R8e.p0o1r3t ultrastructural differences between the hepatocytes treated with 0 (control) and 0.4 mg/kg/day. 1.6 mg/kg/day. For animals given 1.6 mg/kg/day, the average number of peroxisomes per hepatocyte was 13.0. Figure 4 illustrates a representative hepatocyte from this treatment group. Otherwise, general ultrastructural morphology was similar in high-dose livers compared with control livers. High-Dose (3.2 mg/kg/day). The average number of peroxisomes per hepatocyte (12.6) was higher in the high-dose animals compared with controls (Text Table 4). Figures and 6 illustrate representative hepatocytes from high-dose animals. Differences in mitochondria and cellular membranes were not discernible between treated and control livers. Suboptimal fixation, however, of the tissues precluded detailed evaluation of these: organelles. LUNG 0 mg/kg/day (Control). Lung samples were atelectatic and not inflated with fixative. Therefore, most alveoli and alveolar septa were collapsed with poor delineation of alveolar and airway spaces. Figures 7-9 illustrate some normal cellularity and ultrastructural pulmonary anatomy. Type Il pneumocytes (PN2) were randomly identified lining alveoli, terminal bronchioles, or within collapsed septa. Type II pneumocytes were. identified by the presence of lamellar bodies, which are cytoplasmic vacuoles containing electron-dense laminated membrane-like profiles. Lamellar bodies represent the material secreted into the alveolar spaces as surfactant. Occasionally some lamellar body vacuoles were empty, asifthe lamellar material had washed out or been secreted. Surfactant or lamellar material within airway spaces was only occasionally identified in control lungs or treated lungs. The type I pneumocyte (PN1) is characteristically flattened and lines the alveolar space as a thin marginofcytoplasm. Type I pneumocytes differentiate from type II preumocytes. Many pneumocytes appeared to be cuboidal without distinctive lamellar bodies, and may represent a transition stage between PN2s and PN, especially since these lungs were fetal and had not been expanded with air. Additionally, some cuboidal epithelial cells represented terminal bronchiolar epithelium. Type II pneumocytes and lamellar bodies were randomly identified in these control lungs. Some lamellar-like bodies were also present within capillary lumina, although in the control sections examined no lamellar bodies were clearly identified in the alveolar lumina. 0.4 mg/kg/day. Qualitatively, the lung samples from animals given 0.4 mg/kg/day appeared essentially the same as control samples. A representative sample is shown in Figure 10. Random type Il pneumocytes containing lamellar bodies were identified, and only occasionally some lamellar material was seen within alveolar lumina. Ls 0154 PriAmendiicalArPgauhsoNloo.gy41R5e.p0o1r3 1.6 mg/kg/day. Figures 11 and 12 illustrate some representative ultrastructural features from rat fetal lung samples given 1.6 mg/kg/day. These were considered essentially similar to the control and other treated samples. Lamellar bodies were identified within type IT pneumocytes and, occasionally, within alveoli. e3s.s2emntgi/akllgy/sdiamyil(aHritgohc-oDnotsreo)l,. lUolwt-raasntrdumctiudr-adlopsaetshaomlpolgeys.of Tthheehcioglhl-adposseedlnuantgusrewoafsmost of the alveoli and septa made definitive quantitation impractical. Some lamellar bodies were identified within capillary lumina and only a small amount was seen within alveoli. Figures 13 and 14 illustrate some representative lung fields from the high-dose animals. CONCLUSION Ultrastructural evaluation of liver and lungsofrat pups from dams treated with 0 (control), 0.4, 1.6, or 3.2 mg/kg/day (high-dose) PFOS identified increased numbers of peroxisomes within hepatocytes of rat pup livers from dams given 1.6 or 3.2 mg/kg/day. Quantitation of peroxisomes indicated at least twice the number of peroxisomes in animals treated with 0.4 mg/kg/day and triple the number in those given 1.6 or 3.2 `mg/kg/day. Differences in cellular membranes or mitochondria were not discernible between control and treated animals in the samples examined; however, suboptimal fixation precluded a detailed evaluation of these organelles. NInolsuinggnisfaimcpalnetsu,lttrhaestprruecsteurnacledoifffteyrpeencITespnweeurmeoicdyetnteisfiaenddbleatmweelelnarlbuondgifesrowmacsoenvtarloulaatnedd. treated rats. Random type II pneumocytes containing lamellar bodies were identified in all lungs examined ultrastructurally. Although some lamellar material (surfactant) was morphologically identifiable within alveolar lumina, no significant differences were noted between control and treated samples. SIGNATURE OF AUTHOR SeanSA fames B. Nold, D.V.M., Ph.D, Diplomate, A.C.V.P. LE 19 1444 Date REFERENCES McDowell, EM and Trump, BF: Histologic fixative for routine diagnostic light and electron microscopy. Arch. Pathol. Lab. Med., 100:405-414, 1976. . 00011.55!50 IL Light Microscopy and Transmission Electron Micrograph Interpretation Forms ' ! Forms are typographically corrected versions of signed/dated raw data sheets maintained in archived study file. 001554 LIGHT MICROSCOPIC EVALUATION Study No.: 418-013 (PAI99-11 Species/Strain: Crl:CDBR VAF/Plus Rats, Term Fetuses Animal No:_see below Sex: nla Interpreting Pathologist (signature and date): Tissue: __ Liver T Animal | Dose Number Microscopic Findings and Comments mgs }| 1950+-3 ||12.. ECxiaymedtulioacrlyephaermilantgoadpiofifseussiems,m(oEideMrEca,tediffuse,moderate be 155051 13. :MEiMtoHt,icdifiugsuere,smmoidneiramtael mulifoeal el 65064 112.,CyEtMoHp,ladisfmfiucsec,lemaordiengr.atdeiffuse moderate | 2. Cytoplasmic clearing, diffuse, moderate 950571] |13..EEosMiHn,opdhiiflfiusce,grmanoudlearriattye. cytoplasmic,increased.periportal, minimal _| 12. Cytoplasmic leaing, diffuse, moderate b T9516:3 t 1. EEoMsHin.opdihfifluisceg,rmaonduelraaitye,cyt. oplasmicincreased.peiporial,minimal__ 23.. CMiyttte foigupresl,amraiinsnim,admli,fifmusucei,fomcodselrate Oimgk|s| 1951 ||21.. ECVytIo.plGasfmfiscec,lemaoridneg,rdaicffuse, minimal 1653637 137:EEoMsiHn,opdhiiflfiucsger,anmuoldaerriattye.cytoplasmic increased, diffuse, minimal _____ | 2. Cytoplasmic clearing diffuse, light 55634173: EEosMiHn,odpihfifluisceg,ramnouldaerriattye, cyoplasmic inressed. periportal,sight 2. Cytoplasmic clearing, diffuse. light 1554611.3: EEoMsHin,opdhiiflfuisce,gamnoudleraaitye, cytoplasmic increased: perioral light ___ | ___|132.ECoystionpolpahsimliicc cgrlaenaurlianrgi,tdyi.ffcuysteo.psllaisgmhitc, increased. diffuse.minimal ____| T| 1586 21.. ECMHy,doiffupsec.llemaordiaengr,adseilfyfm-usseevi.selriecght |43. EMiotsoitniocphfiilgiucrers,anmuilniamiayl., cmyutloipfloacssmlic, increased, periportal light Page 1063 00155) Dose T| NAummbaelr | MicroscopicFindingsand Comments Tomgig || 19560-1 || 1. Edoismuosphilc granularity,cytoplasmic, increased, moderate, ee1n 7165m 03 m B1.o EEMMm EH,,Gdifife usse,,mMoOs dGeErTaRtEeNi lYySseVvere Ee. oes || 132.. ECoystionpolpahsimliiccgrlaenaurlianrgi,tdyi,ffcuysteo,pmlaisnmiimca,lincreased,diffuse, slight sss 95803 1.MEUMHO.diSfEfUuEsReS,:mFoUdNeNrEaLie, EUIOCHL eee] i 1123.. CEoystionpolpahsimliicccgrlaenaurlianrgitdyi,ffcuysteo,pmloadsemriact,ei:ncreased. periportal, minimal 1552.2] 41:. MEiMtHot.idcifgfuursee,s,momidneirmaatlclym-usletviefroecal | | 2. Cytoplasmicclearing, diffuse light m1| s 718m 562. 311.23.: EECoMystHion,polpadhsiimflificsccegl,reamanrouidlneagrr,iadtciylf,yfc-uyssteeov,pmeliraensmiimca,lincreased.diffuse. slight. . | |3. Eosinophilicgranularity,cytoplasmic,increased,diffuse, slight Tamgks |i 193683 | 1. Edcisfifmusoephihe granary, cytoplasmic, imereased, moderate, | 2. EMH,diffuse,moderatly-severe: 163657 |13:EMoistiontopihcifliigcuressr,asnluilghatiymu,lctiyfoctaloplinacrseasmedi.sc_g_h,______________ 2. EdiMfHfu,sdeiffuse, moderately-severe ee TSSTET 3| :1. MEiotsoitnioepfhiigluircesgr,asnluilgahrti,tmy,uclytiofploascmaicl. inc__r.e.a.s.e..d.,..m.o.derate, difgrfuse 2. EMH, diffuse, moderately-severe OI OO1957741...Eos.ino.phiF lic granc ularityB .cytoplasmie c. increased, moderate, , | 2. EdiMfHfu,sediffuse.moderatseevlerye T6o57o5,3 1. MEBoOsinopShiElUicSSgraInulNarSity,. cytopOlaSsmHic. increased.moderate diffuse | 2. EMH,diffuse, moderatly-severe Page20f3 01553 Tncidence and Severity of Light Microscopic Liver Findings Microscopic Finding PFOSDose Levels (mg/kg/day) [) 04 16 32 mie 2 [0] wmercomned]O _o| `Eosinophilic granularity, cytoplasmic, increased minimal| 2 2 1 slight| - 3 3 moderate| - - 1 Cytoplasmic clearing minimal| - 1 2 slight| - 4 1 moderate| _5 - 1 Extramedullary hematopoiesis moderately-severe| _- 1 4 `Mitotic figures, increased minimal| 2 1 2 2 slight | - - 2 Page 3of 3 001554 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: _418-013:EM-0939 Animal No: __19504:3 Treatment Group: _0 mg/kg, Control Sex: _nonapplicable Species/Strain:C.CDOBRVAEPLs Rat Tissue: Liver Block Nofs).: __99.39-1 Photo/Negative No(s):ZZ15I7S7T88-2 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): 3.10.99 Features: ZISTIE. Liver. Hepatocyte adjacent to central vein a1 and bottom. Bile canaliculus at top. Cytoplasmic organelles are clumped together with intervening spaces of fine granular material. Mitochondria appear dense and have some dilated cristae. Five (5) peroxisomes counted. ZI5779. Liver. Hepatocyte exhibiting similar cellular details as Z15778. Zero (0) pZeIrSo7x8i0s.omLeisvecrl.eaHrleypaitdeonctyiftieeda.ppears similar above examples with clumped organelles lAowheermaetfotp.oiMeittiocchceolnldirsiianatphpeeatorpdreingshet sainndushoaivdeasnodmaebdiileatceadnaclriicstualeu.s iTshpereese(n3t) in the. ZpIeSrTo8xIi,somLeisvecro.uHnetepda.tocyte and adjacent sinusoid containing two nucleated rd blood cells. Mitochondria appear dense with some granular deposits and dilated cristae. Few numbers of peroxisomes identified, although poor fixation makes their clear identification dZiIfSf7ic8u2l.t. LFiovuerr.(4)Cepnetreorxihseopmaetosccyotuentiesds.urrounded by several other hepatocytes. wth a sinusoid containing a nucleated erythrocyte in bottom right anda bile canaliculus in top portion of photograph. Mitochondria appear dense with some granular deposits and dilated cristae. Four (4) peroxisomes counted. Conclusions: Normal hepatocyte. Suboptimal fixation. Average 3.2 peroxisomes per hepatocyte. 001559 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013; EM-99.39 Animal No: ___19504-3 Treatment Group: _0 mg/kg, control Sex: _a_p nop n licable Species/Strain: Cr.CDBR VAF/Plus Rat Tissue: Lung Block No(s). 99.392 Z15783 - Photo/Negative No(s).:_Z15787 Significant Lesions (check one): Yes X_ No Intepreing Paologit (signature and date): TFeaamtulraesb:ccZn157A83d. eLeungi. nDarcklllinrineg cceolblorisdaaltywpiehIlsphnoenumdoacyrtea(nPiN2)mwehirch conthaeins four cncydotopCpumlalaslmsweciotnhsiasteonbtlwointghlgleyecosgena.dSaubbujnadcaenntt pto ithke seurfmacae iisla cianpi1l5laerpycaonpdlaemnd,opthreolpiaabllycelal SnZ1a5n5ru2 r Lmratneeg.riiClemnptraiolbdlaeyrlkglli iynrtioneggeebnlassswlalo.PgNoZnD.owirtAhadlsiesv1caernnatl illnaimoneglclcalcbsocdcioewsni.ttihnAmsilbmaoerrgneoduaasnraBroef amacchroopnhaagre.i aAn<d saame epannddoipetnadasomthyetadceull i an0dhseerilghtdofnhceBPrNa.nle, proba ZLTTE Lar, hone of oud sph cls wht sat granteemater itheey CE hoveerneagen out. Subscene 3 calla ning endodelo cel 3d 5 ZI 1578, Lung. An locus wit several cubed ping ces Appear o be unflatened rciigm1g5ae2e8s5livaelLooncgofa.tsFANr.eTohfecollolnagpsledeavperanodbcaapanilrlbafrilsesry.. ErDythreocytisessawicathoiniycraalcan.soudple a Conclusions: Normal lung. Few type II pneumocytes and lamellar bodies identified. 001556 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.:_418:013; EM-99.39 Animal No.: __19505-| Treatment Group:_0mg/kg, control Sex: _non applicable. Species/Strain: CrLCDOBRVAEPlus. Rat Tissue: Liver Block No(s): 99Z3IS7988 -3 Photo/Negative No(s).;_Z15792 Significant Lesions (check one): Yes __X_ No Interpreting Pathologist (signawre and date: ____ Features: ZIST88. Liver. Hepatocyte contains abundant fine granular material and some clumpingofmitochondria and endoplasmic reticulum. Mitochondria are dense and contain some dilated cristae. Few peroxisomes are noted, four (4) counted. Z15789. Liver. Hepatocyte appears similar to Z15788, with fine granular cytoplasmic `material and clumping of larger organelles. Four (4) peroxisomes counted. ZI5790, Liver. Hepatocyte. Ventrally is a large sinusoid witah Kupffer cell among Several erythrocytes. A hematopoietic cell is on the lower right. Some dilated blebs of rough endoplasmic reticulum (RER) are evident in the center hepatocyte. . Mitochondria appear dense with some granular deposits and dilated cristae. One (1) peroxisome counted. ZIS791. Liver. Hepatocyte appear partially detached from adjacent cells and. suboptimally fixed. . Mitochondria appear dense with some granular deposits and dilated cristae. Two (2) peroxisomes counted. Z15792, Liver. Hepatocyte appears essentially similar to those described above. `Abundant fine granular material in cytoplasm.. Mitochondria appear dense with some granular deposits and dilated cristae. Some dilated blebs of RER noted. Four (4) peroxisomes counted. Conclusions: Normal hepatocyte. Suboptimal fixation. Average 3.0 peroxisomes per hepatocyte. 001554 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013.EM-0939 Animal No: __19505-1 Treatment Group:_0mg/kg, Control Sex: __nonapplicable Species/Strain:CRaitCDBR VAF/Plus Tissue: Lung Block No(s). 9__93219579-3 -4 Photo/Negative No(s).:_Z15797 Significant Lesions (check one): Yes __X No Interpreting Pathologist (signature and date): Features: Z15793, Lung. Alveolus with lining cells. Two cuboidal pneumocytes with fine granular cytoplasm. Top cell has an extended flatened portion that contains a lamellar-like structure, and may be a PN2 that is becoming a PN1. Normal, more flattened PN1s are present adjacent to these two central cls. 15794. Lung. Two of four cuboidal lining cells have lamellar bodies characteristic of PN2. Smaller and darker cells has a microvillous border than contains a lamellar structure, probably alveolar surfactant. Z15795. Lung. Alveolus with lining cells. Dark cuboidal cell is a PN2 and contains several lamellar bodies. Some flattened PNs are present as well as another cuboidal preumocyte. A capillary containing erythrocytes is within the alveolar septum. 215796. Lung. Alveolus and areaofcollapsed septa. Two PN2s are present, both containing two to six lamellar bodies. No alveolar luminal lamellar structures are: evident. Z15797. Lung. Alveolus and lining cells. The dark columnar cell is a PN2 and contains Several lamellar bodies. The left adjacent pneumocytes appear not to have flattened out as more differentiated PN1s. A second PN2 is just to the right of the central PN2, and contains several small lamellar bodies. Subjacent tot he alveolus is a capillary containing a leucocyte and endothelial cell Conclusions: Normal lung. A moderate number of type 1 preumocytes were identifiable, and contained lamellar bodies. 0155% TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013; EM-99.39 Animal No.: _19506-4 Treatment Group: _0 mg/kg, control Sex: __non applicable Species/Strain: CL.CDBRVAE/Plus. Rat Tissue: Liver Block No(s). __99.39-5 Z15808 Photo/Negative No(s).:_Z15812 Significant Lesions (check one): Yes __X No Interpreting Pathologist (signature and date): ____ Features: ZI3808. Liver. Hepatocyte contains abundant fine granular cytoplasmic material and some clumping of predominantly mitochondria and rough endoplasmic reticulum. Five (5) peroxisomes counted. Z15809, Liver. Hepatocyte appears similar to Z15808. Some dilated blebs of RER are noted. Six (6) peroxisomes counted. Z15810. Liver. Hepatocyte has a washed-out appearance to much of its cytoplasm, and is surrounded by hematopoietic cells ventrally. Endothelial cells are incompletely defined. At the top leftof the photograph is a bile canaliculus. Two (2)peroxisomesare counted. ZIS811, Liver. Hepatocyte with prominent bile canaliculus at lower left and sinusoid on right margin of photograph. Mitochondria are dense and contain some dense deposit and dilated cristae. Six (6) peroxisomes counted. Z15812. Liver. Hepatocyte contains abundant washed-out cytoplasm and clumping of RER and mitochondria. At the top are two hematopoietic cells and a bile canaliculus is present in lower right of photograph. Three (3) peroxisomes counted. Conclusions: Normal liver, Suboptimal fixation. Average 4.4 peroxisomes per hepatocyte 001554 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _41. ; EM-99.39 Animal No.: _19506-4 Treatment Group: _0 mg/kg, control Sex: non applicable Species/Strain: Cl.CDBRVAF/Plus Rat Tissue: Lung Block No(s). 99.39-6 Z15818 - Photo/Negative No(s).:__Z15822 Significant Lesions (check one): -- Yes X No Interpreting Pathologist (signature and date): Features: ZISS1E, Lung, Dak coll with round mcleds Tn te comer 3 PNG which pacally lcisiyntieomspmtlehdaeisaamltrveeelpoyrleaasrdejsnaptcasceena.tn taShdeejvaPecNre2an.lt lPTaNmhe.llraAercmbaaoipdniidelesrrayorfechoweintahtliinisunietgs1casynCtoeolfpllyaafpsrsmoe.cdyatTlehveewoilltiahriagnnedtchlseeeaspreaptum fZ1h58e19p,hoLruongga.pAht.Thwione-ndmotaherolifgaPliNnIcoslsuaprperaao0pruiotnnruhddeinsGgaspeipratruym.awhnidchcacopnitalnlisnaatrnhye middle a aerytchroomcyatle.boToomthe15 arg right is a(spoeme| wphreautmdoacrykteer,PwNi2t,h whliacrhgecpoonltyaignosnaslevneurcalleulasmaenlldarmobsoddileys.desIen ZIS820, Lung. PN1s line several sepa inthis photograph. Dorsal, the septal capillary ZceoIvnSitSda2ei1nn.ts aNLolunydgmi.spcheoArcnlyitvbeleeasnPdaN2nasnluoicfnlielnaagmteecldellealrrsby.tohd&rioePcsyNat2er.eis Icpnirtdeheseencntenitneirncga pillary a th alveol granulocytic us. I is th cell is darker imilssiwnigh. sTewveoraPl Idsieardc meveidlenat obnodtiheersi.ghMtosmtarogfitnhoefamnieslallrvemoalatresrpiacei.n hLeasregebonduicsleuiss on the ZTeItSaEp5p.eaLrusngo. beAllovnegooluasnanedndsoetvheerlailalincilngHcielnlg. aMseesptaapcpaepaarr1y6,be PN1s with abundant cSvoimdeewnh,aatltchlaosueghyotpwlsamsalmlwvhaiccuhohlaesnn'tacnomipnlgetperleyalmaotceynteedaowh.e Nboordieofminoiftihvee PpNho2tsogarreaph omfatyherepphroetsoegnrtaplha.mellar bodies. A capillary omauning two mucleaed cells occupies the High side Conclusions: Normal lung. Occasional type If pneumocytes containing lamellar bodies identified. No lamellar material noted within alveolar spaces. 001560 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013: EM-99.39 Animal No.: __19509-1 Treatment Group: 0 mek, control Sex: __non applicable `Species/Strain:Cr.CDBR VAF/Plus Rat `Tissue: Liver Block No(s): __929.1359:8723 Photo/Negative No(s).;_Z15827 Significant Lesions (check one): Yes _X_ No Interpreting Pathologist (signature and date): Features: Z15823. Liver. Hepatocyte bordered by sinusoid containing nucleated erythrocyte on left and other hepatocytes on bottom and right. Anendothelial cell is in the upper left portionofthe photograph. There is clumping of the mitochondria and RER `with an intervening open area or areas filled with fine granular material. The RER is dilated. Mitochondria contain some granular deposits. Five (5) peroxisomes counted. 215824, Liver. Hepatocyte. At the top is a nucleated erythrocyte. Sinusoidal endothelium is poorly delineated. The RER shows some blebbing and dilatation, and mpietroocxhiosnodmreisachouanvteeds.ome dense deposits and some dilatationofcristae. Five (5) Z15825. Liver. Hepatocyte surrounded by hematopoietic cells. The RER and nuclear envelope shows some dilatation and blebbing. Mitochondria appear dense and have some `granular deposits and dilated cristae. Seven (7) peroxisomes counted. 215826. Liver. Hepatocyte appears essentially similar to above described cells with clumping of major organelles. Five (5) peroxisomes counted. 215827. Liver. Hepatocyte surrounded by hematopoietic cells. A single larger vacuole is present in the cytoplasm and contains a flocculent like material. Four (4) peroxisomes counted. hCeopnactloucystieo.ns: Normal hepatocyte. Suboptimal fixation. Average 5.2 peroxisomes per co1s6 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013; EM-99.39 Animal No: _19509-1 Treatment Group:_0mg/kg, control Sex: __non applicable `Species/Strain: CiL.CDBBRVAF/Plus. Rat Tissier Lung Block No(s): _9_ 939-8 715828 Photo/Negative No(s).._Z15832 Significant Lesions (check one): Yes X__ No Interpreting Pathologist (signature and date): ____ cFoenattauirness:sevZe1r3al82l8a.melLluanrg.bodiAelsv.eoAluns earnydthlrionciyntgeceilslis.mmTehdieadtaerlkyertocuthbeoildeaftlocfetllheisPaNP2Na2ndand appears to be within the alveolus. At top let is a pneumocyte which has two small lamellar-like bodies and is probably a PN2 in transition to a PN1. A flattened PN1 is immediately on top and to the right of the dark PN2. Beneath these cells the septa are collapsed together. 215829. Lung. Alveolus and lining cells. The smaller dark cel is a PN2 with several lamellar bodies. To its right is a flattened PN1 and to is lefts a cuboidal PN. The lamellar or myelinlike material o the rightofthe PN2 appears to be within a capillary lumen. At the bottom right ZisL5a8t3h0i.n-wLaulnlge.d seTpwtoumercytohrnoctytaesiaacrnaepiiwlilntahrgiyn atnhedcearpyitlhlraorcyytleu.men. The lining preumoeytes are Tattened and appear to contain several vacuoles that may have contained lamellar bodies. Subjacent are collapsed septa and 2 mixture of septal cells including pneumocytes and eZnIdSo8t3h1e.liaLluncge.ll. Areaofcollapsed alveolar septa. Atthe left middle and right middle-top are ark cells which contain lamellarbodiesand are PN2s. Other cells are compressed pneumocytes, endothelial cells, andbloodcells within capillaries. Some lamellar debris appears to be within capillary lumina. 215832, Lung. Area of collapsed septa. The large round clear cell is a cuboidal pneumocye. tDhoerslaulmteontohfistahreecaepnidloltahrye.lial cells lining a capillary. The lamellar structure appears o be within Conclusions: Normal lung. Scattered type Il pneumocytes present. Some lamellar material also present within capillaries. No lamellar bodies noted in alveolar spaces. 00156), TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013; EM-99.39 Animal No: _1_ 95162 Treatment Group:_0mg/kg, control Sex: __non applicable Species/Strain:CilCDOBRVAF/Plus Rat Tissue: Lung Block No(s): 9939-10. Z15838 Photo/Negative No(s).:_Z15842 Significant Lesions (check one): ___ Yes _X No Interpreting Pathologist (signature and date): Features: Z15838. Lung. Two pneumocytes a the bottom are PN2 and contain some lamellar bodies. Dorsal to them are collapsed septa containing primarily pneumocytes and endothelial cells, 215839. Lung. Several pneumocytes are present. Central pneumocyte contains several lamellar bodies and is a PN2. Flattened PN1s line the protruding septum which contains acapillary, endothelial cell and leucocyte within the capillary. Z15840, Lung. Single septum lined by PN1s on either side. On the right top is a PN2 with several lamellar bodies. The right lower pneumocyte also contains a single lamellar body. The septal capillary and endothelium are also evident. ZIS841. Lung. Area of collapsed septa. At right are two pneumocytes which contain multiple lamellar bodies. Other compressed septal cells appear to be pneumocytes and endothelial cells. 215842. Lung. Area of collapsed septa. Several cuboidal pneumocytes are present . Centrally is a capillary containing an erythrocyte. Next to tis a couple lamellar structures that appear to be within the capillary lumen. Conclusions: Normal lung. Random type Il pneumocytes containing lamellar bodies were identified. No lamellar bodies were clearly identified withinalveolar spaces. 0015:63 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: 418-013, EM-9939 Animal No _195162 Treatment Group: _0 mg/kg, control Sex: __non applicable. Species/Strain:RCaALCDBBR VARPlus Tissue Liver Block No(s) __ 9939-9 715833 Photo/Negative No(s). Z15837 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): Features: Z15833, Liver. Hepatocyte with a sinusoid al lower right and a bile canaliculus on the lef. Cell has abundant washed-out cytoplasm and clumping of mitochondria and RER. There is dilatation of RER and the nuclear envelope. Mitochondria contain some granular deposits and some dilated cristae. Plasma `membranes and sinusoidal endothelial cells are poorly delineated. Five (5) peroxisomes counted. 215834, Liver. Dilation and blebbing of RER. Some granular deposits present in mitochondria. Nine (9) peroxisomes counted. 215835. Liver. Mitochondria contain some dense deposits and dilated cristae. There is blebbing of the RER. Plasma membranes are poorly defined. Five (5) peroxisomes counted. Z15836. Liver. Hepatocellular morphology essentiallysimilaras those described above, Four (4) peroxisomes counted. Z15837. Liver. Mitochondria contain some dense deposits and dilated cristae. Plasma membranes are poorly defined. A multivesicular body is present to the right of the nucleus. Cell at lower left appears to be an endothelial cell, but may be a hematopoietic cell in the sinusoids. Two (2) peroxisomes counted. Conclusions: Normal liver. Suboptimal fixation. Average 5.0 peroxisomes per hepatocyte. 001564 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 Animal No. __19534- `Treatment Group: _0.4 mg/kg Sex: _non applicable `Species/Strain:CiCDEBRVAF/Plus Rat Tissues __Liver Block Nos): __99.39-31 Z15935 - Photo/Negative No(s).._Z15939 Significant Lesions (check one): X Yes No Interpreting Pathologist (signature and date): Features: Z15935. Liver. Hepatocyte with poorly-defined plasma membrane and no surrounding endothelial cells. Some hematopoietic cells are around this hepatocyte. Mitochondria contain some granular deposits and some dilated cristae. RER is dilated and shows some blebbing. Four (4) peroxisomes counted. Z15936. Liver. Hepatocyte shows marked dilationof RER and poor delineation of the plasma membrane. The nuclear envelope is also dilated. A large clear vacuole is present in the right side of the cell. Eighteen (18) peroxisomes counted. Z15937. Liver. Hepatocyte with clumping of organelles and intervening washed-out areas of cytoplasm. Mitochondria contain some granular deposits and some dilated cristae. RER is dilated and shows some blebbing. Eleven (11) Peroxisomes counted. Z15938. Liver. Hepatocyte shows similar ultrastructural morphology as those described above, with suboptimal fixation. A multivesicular body is present to the right of the nucleus. Eleven (11) peroxisomes counted. Z15939. Liver. Hepatocyte with clumpingof organelles and intervening washed-out areas of cytoplasm. Plasma membrane is indistinct. A hematopoietic cell is at the bottom of the photograph. Mitochondria contain some granular deposits and some dilated. cristae. RER is dilated and shows some blebbing. Sixteen (16) peroxisomes counted. `Conclusions: There appears to be increased numbers of peroxisomes per hepatocyte Scobmoppatriemdalwith control liver. Average 12.0 peroxisomes per hepatocyte. Cellular fixation c0156% TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: 418-013EM-9939 Animal No: __ 1953-1 Treatment Group: 04mg/kg Sex: __nonapplicable Species/Strain:RCa.rCDBRVAF/Plus Tissue: Lung Block No(s): _992.1539941--32 Photo/Negative No(s).._15945 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): pFneeatuumroecsy:teZsI5(9P4N1I).LuBneg,neaAtlhvetoheluPsNaInsdilsainirnogucnedllst.o pAolllygthoenailnicenlglcceolnltsasipnpiengar5tolabmeeltlyarpbe|odies in of tsescctiyotnopilsassomm;ehwihsaits f2lattytpherIofupgnhetuhmeosceypttael(wPaNl2,). This s probably a lining cell, and the plane 215942. Lung. Alveolarspaceandlining cells. The dark and somewhat flatened lining cell is a PN2 and contains several vacuoles empty of contents. These appear to be "empty" lamellar bvoadciueosl.es.The lightr-staining pneumocyte at the far left also contain two similar-appearing 2bu1t59a4l3s.o atLtuancgh.esAwlivtehotluwso aanddjalciennitngmcoelrles.lucTehnet PdNa1rsk.nucNloeulsamieslplaarrtbofoadiefslantotteendedipnntehiusmocyte, photograph, although a vacuole adjacent o the op lef nucleus may be an "empty" lamellar 2b1od5y9.44.SepLtuanlg.walTlhseadnadraklvleionliingarceecololnllaptshedriagnhdt cnoonrtmaainlsspeuvlemroanlarlyamaerlclharitbeocdtuireesisanidnidisstainPctN.2. "iTnhteheotchhearralcitneirnigstcielclst,ypaeltIhponueghumcoucboyitdealf,asahpipoena.r to be preumocytes which have not flattened out 215965. the septal Lun. Alveolus and wall and are PNIs. lining cell. No lamellar bTohdeietswoaredasrekenliinnintghlesleslibnoitnhg caeplple.arTfhaetiteonpecdelalloinng, tihdeenptihtoytoisgruancplheaarp,pebaurtsmdaeytabceheadmaancdrospurhraoguendoerdenbdyotshoelmiealexctelrla.celAluclaaprillliapirdy-lisikpermeasteenrtiaalt. thIetstop lef, Conclusions: Essentially normal fetal lung. Random type Il preumocyies with lamellar bodies identified. No remarkable abnormalities noted. 001566 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418:013 EM-99.30 Animal No: __195363 Treatment Group: _0.4 mg/kg Sex: __nonapplicable Species/Strain:RCaLtCDOBRVAE/Plus Tissue: Liver Block No(s).: 99.39.33 Z15946 - Photo/Negative Nofs).._Z15950 Significant Lesions (check one): __X_ Yes No Interpreting Pathologist (signature and date): Features: Z15946, Liver. Hepatocyte. There is dilatation of RER and of the nuclear envelope. Some granular deposits are evident within some mitochondria. Four (4) peroxisomes counted. 215941. Liver. Hepatocyte. There is some clumping of organelles. The RER and nuclear envelope are dilated. Seventeen (17) peroxisomes counted. Z15948. Liver. Hepatocyte. There is poor definition of the plasma membrane. RER and the nuclear envelope are dilated. Fourteen (14) peroxisomes counted. 215949, Liver. Hepatocyte appears similar to those described above. Eighteen (18) peroxisomes counted. Z15950, Liver. Hepatocyte. At the top is an endothelial cell and several adjacent erythrocytes and one nucleated erythrocyte. The plasma membrane of the hepatocyte is indistinct and organelles are clumped in the cytoplasm. RER is dilated. Eight (8) peroxisomes counted. Conclusions: Increased number of peroxisomes per hepatocyte compared with control samples. Average 12.2 peroxisomes per hepatocyte. Suboptimal fixation. co1559 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.:_418-013EM-0939 Animal No ___ 19536 Treatment Group: __0.4 mg/kg Sex: __non applicable `Species/Strain:RCart.CDBR VAF/Plus Tissue: Lug Block No(s): __99.39-34 Z15951 Photo/Negative No(s).:_Z15955 Significant Lesions (check one): Yes _X No Interpreting Pathologist (signature and date): dFeebartiusreisn:cluZdIiSn9g3sLo.meLulnagm.ellAalrvemoatleursiaalndcoanrseiasotfecntolwliatphsseedcrseetpetda.laTmhelelabrotbioodmieasl(vseuorlfuasctcaonntt)ains ifdreonmtiPfiNe2ds.. AtThleeasltintiwngopcnapeiulmloarciyetseasrearpereasllenftlaitnttenheedcaonlldaappsepdeasrepttoalbearPeNa.1s. No PN2s are 215952, the right tLhuenngu.clAeluvseaonlduscealnldbelnineiantghcetlhles.PNPN1cosntlainientshaesailnvgeloeladrarskpalcaemealnldarabreodfyla,ttwehniedc.h On ascuagpgielsltarthyisancedllsommaeybneonaidPeNn2t.ifiOetdhceerllsc.ellNs oinltahmeelplhaortboogdriaepshaarree pernedsoetnhteliinalthceelallsv,egorlaanrulluomceynt.e in 215953. is within Lung. The prominent dark lamellar or a capillary space. Type | pneumocytes myelin-lke body ar flattened and in the center of the photograph line the alveolar space. At the bboodtiteosm, aofPtNh2e.phNootoagbrnaoprhmailnitthieesconloltaepds.edseptal area is 2 cell which contains two lamellar 2a1nd59i5s4a,PNL2u.ng.ThAelvaedjoalcuesntancdellliinsincgubcoelildsa.l,Abustmaalplpecaernstrtaol bdear2kpcrelelucmoonctyatiensw3h-i4chlahmaesllnaortbodies pfrlaotmtienneedntouti.n tAheccaepnitlelroarfytchoentpahiontionggrasepvhe.ralSoemreytdhebrroicsyiastneprdseasenltiniinngtheendaoltvheeolliaarllcuemlleni,s and some may be fine lamellar material. and is also a PN2. The nucleated cell at the top contains two lamellarbodies 215955. septa fill Lung. the top Lamellar bodies ofthe page. No awbintohrimnaaliPtNie2s lnionteedt.he alveolar space. Collapsed alveoli and Conclusions: Essentially normal fetal lung. Some type Il pneumocytes identified containing lamellar bodies. Some lamellar bodies present within alveoli. 001563 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 Animal No: __19543-4 Treatment Group: _0.4 mg/kg Sex: __non applicable `Species/Strain: C.CDOBRVAFPlus Ra Tissue: Liver Block No(s).: 99.3935 215956 - Photo/Negative No(s).:_Z15960__ Significant Lesions (check one): _X__ Yes No Interpreting Pathologist (signature and date): Features: Z15956. Liver. Hepatocyte with clumpingofRER and mitochondria. Plasma membranes are indistinct. Cross-sections of RER appear dark. A bile canaliculus is present at the top right. Several hematopoietic cells are around the periphery. Mitochondria contain some dilated cristae. Eight (8) peroxisomes counted. 215957. Liver. Hepatocyte morphology appears similar to Z15956. Organelles are clumped in the cytoplasm; intervening spaces contain fine granular material which is probably glycogen and cytosolic debris. Five (5) peroxisomes counted. Z15958. Liver. Hepatocyte with bile canaliculi present at top and bottomof cell. Fifteen (15) peroxisomes counted. Fixation is very poor. 215959. Liver. Hepatocyte with marked clumping of organelles. Two hematopoietic cells are present at the bottom of the photograph. Three (3) peroxisomes counted. Z15960. Liver. Hepatocyte surrounded primarily by hematopoietic cells. Sinusoidal endothelial cells are absent due to poor fixation. Hepatocytic mitochondria contain some granular deposits and dilated cristae. Nine (9) peroxisomes counted. `Conclusions: Section of liver exhibits suboptimal to poor fixation. Numbers of peroxisome per hepatocyte only very slightly higher than control values. Average 8.0 peroxisomes per hepatocyte. o0156 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: 415-013EM-9930 Animal No: 195434 Treatment Group: _0.4 mg/kg Sex: _nonapplicable Species/Strain:CCD@RRVAFPUs Rat Tissue: Lung Block Nof(s).: _99.39-36 Z15961 - Photo/Negative No(s).:__Z15965 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): ______ aFetaetrumriensal: bZrLo5n9c8h1i.ole.LuTngh.ecAelnvteroallucsubaoniddaflmicenlglcceollnst.aiLnsinsienvgercaellllsaampelpleaarrbcoudbioeisdwailthainndamfaiynebc agpriacnaullavraccuyotloepsl.asSmoicmebalcakmgerloluanrdm.atOerniailtihs eprseisdeentairn ttwhoadlavrekoelrarcospmapcree.ssed cels which contain Con1t5a9i62n. abLuunndga.ntSgervaenruallacrumbaotiedrailalpnineutmheoicryctyetsopllianesmt,hiwshaiicrhwaiysocronaslivsetoelnatrwsiptahceg.lyTcohgeeyn.alNo cPaNpi2lslaarrieesevairdeenptr.eseAnt,floanteendeodrsPalNly aisndproesneentveantrtahlely.ighBtotthopcoofntthienaslvoemoelalrasmpealclea.r mTyweloinse-pitkael Zm1a5te9r6i3a. iLnutnhgei,r lPuhmoitnoag.raph shows several cuboidal pneumocytes, th top cll of which contains ccalpeilalrarvyatcouotlhee sleufgtgoefsttihveeloifnainng c"eelmlsp.tyT"hleamceelllasrtbtohdy.botAtodmarlkeftercyotnhtraoicnystae ifsepwressmeanltlilnatmheellar 2b1o5d9i6es4,andLusngp.roAblavbeloylausPNa2n.d lNinoinlgacmeellll.arTbhodeielsininnogtecdellinsaarlveoplrie,daonmdinnaontalbynofrlmaatltienteidesanndoted caepnpteraalr tloinbiengdcieflfl.ereTnhtieatdeadrPk vNa.cuoSleevemraayl rmeiptroecsheonntdaridaepaletaneddaldarmkelvlaacrubooldey.areTphreesdeanrtknlatmheellar 2m1y5el9i6n5l.ikLeunsgt.ructTuhree nceelalratthethteopboitstwoimtohfinatcheappihloltaorgyraspphacceonwtiatihnascoslevlearpasleldamseelpltaurmb.odies and is cAoPntNa2inwiintghidnartkh ercyotlhlraopcsyetdesseaprteubme.nePathNthelisneutrfhacaeierpsipthaecleioumn.thNeoriaghbtn.orTmawloiticaepsilalearineosted Conclusions: Essentially normal fetal lung. Several type Il pneumocytes identified with lamellar bodies. Also, some lamellar structures noted within alveolar spaces. co15pp TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: 418-013EM-9939 `Animal No.: 19546-1 Treatment Group: __0.4 mg/kg Sex: __non applicable Species/Strain:Cr.CDBRVAF/Plus Rat `Tissue: Liver Block No(s). 99.39-37 Z15966 - `Photo/Negative No(s).:__Z15970 Significant Lesions (check one): ___X Yes Interpreting Pathologist (signature and date): ____ No Features: 215966, Liver. Hepatocyte. Plasma membrane is indistinct and cytoplasmic organelles are clumped together. Mitochondria show some dense deposits and dilated cristae. Fifteen (15) peroxisomes counted. Z15967. Liver. Hepatocyte appears morphologically similar to Z15966, with clumping of organelles, indistinct plasma membranes, and mitochondrial changes. RER appears dilated. Seven (7) peroxisomes counted. Z15968. Liver. Hepatocyte. A prominent bile canaliculus is present in the bottom right corner. Organelles are clumped with large intervening spaces of fine granular material, Mitochondria show some dense deposits and dilated cristae. Twenty (20) peroxisomes. counted. Z15969. Liver. Hepatocyte appears essentially similar to Z15968. Fixation is poor. Eleven (11) peroxisomes counted. Z15970. Liver. RER is dilated and plasma membranes are indistinct. An endothelial cell Tiswneoltvepr(e1s2e)ntpaetrsoixniussomatestocpo.unMtietdochondria show some dense deposits and dilated cristae. Conclusions: Liver exhibits poor fixation. There appears to be increased numbers of peroxisomes per hepatocyte. Average 13.0 peroxisomes per hepatocyte. cos `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 Animal No: _10546-1 Treatment Group: _m0g .4 /kg Sex: __non applicable. Species/Strain: Crl.CDOBRVAF/Plus Rat Tie: Lung Block No(s). 99329 159-713- 8 Photo/Negative No(s).:_Z15975 Significant Lesions (check one): Yes _X No Interpreting Pathologist (signature and date): _____ Flaemaetlulraersb:odZiIe5s,97m1o.stLpurnegd.omAilnvaenotlluystahnedcelinntienrgcecellllsw.itSheavterlaealstlinniinneg clealmlesllcaorntbaoidnies. This is a distinctive pneumocytes whose PN2. The three cells at alveoli are out of plane the top of of section the photograph appear orare collapsed. to be 215972. alveolar Lung. space. Photograph Both sidesof slightly out the septum of focus. are lined An unattached PN2 by flattened PN1s. is A present in the capillary and endothelial alveoli. cell are evident within the septum. No lamellar material is present in the 215973. "empty" Lung. Alveoli lamellar bodies. and septum. The The other lining dark cells cell are on the PN1s. top left is a A capillary PN2 with with several a luminal erythrocyte alveoli. is present in the center of the septum. No lamellar material is present in the 2th1e59al7v4e,oluLsu.ng.NexAtlvteootluiss aancdubloiindianlg cpenlelsu.moAcyftleatweintehd sPeNveIrallincelseatrhevabcoutotloems,maarPgNi2n.of `dTihleatveadcnuuoclleesarareenv"eelmoppetyanldamdeilsltaernbdoeddieesn.doTphlaescmeinctrraeltcieclullwuimtihs aa mciucbrooivdialllpourseubmorodceyrt,e e2x1h5i9b7i5t.ingLusnogm.e eCluebmoeindtasl olfinsiunbgocpetlilmianlcfeinxtaetriorni.ghtNoanldacmeellllaatrcbeondtieerslnefottebdotihn ctohnetaalivneoolin.e coorntvweortviancgutoolePsNwIhsiacfhtearpepxeparrestsoibneg ltahmeeirlllaarmeboldliaers.(suTrfhaecsteancte)llsmaaterreiaplr.obaNbolylaPmNe2llsar material noted in the alveoli. Cboodnicelsusidieonntsif:iedE.ssenNtoialllaymenlolramramlatfeertialalluinsg.preSsceanttteirnetdhetyapleveIollipneumocytes with lamellar 00157 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 Animal No: _1_95462 Treatment Group: _0.4 mg/kg Sex: __nonapplicable Species/Strain: CALCDBRVAFPlus Rat Tissue: L __ iver Block No(s).: __99.39:39 215976Photo/Negative No(s).:_Z15980 Significant Lesions (check one): ____ Yes X No Interpreting Pathologist (signawre and date): ___ Features: organelles Z15976, Liver. are clumped and Hepatocyte exhibits apoorly-defined plasma membrane. the mitochondria show dilated cristae. At top right is an The hinetmearsttoiptioailecteilcl cseulrlrsowuintdheidn baysicnoulsloaigde.n, Fprooubra(b4l)ypaerfiobxriobsloamste.s cAotunttheed.lower left are some 215977. Liver. Two hepatocytes with an intervening bile canaliculus. Hematopoietic cells are present in the upper right and lower left sinusoids. The larger cytoplasmic organelles are clumped together and the cytosol appears washed-out or contains a fine granular material. The mitochondria show cell) contains seven (7) peroxisomes. some dilated cristae. The hepatocyte (bottom 215978. Liver. above, fixation Central is poor. hepatocyte in photograph appears Two (2) peroxisomes counted. similar to those described 215979. Liver. Hepatocyte, whose morphology is similar to those described above. Five (5) peroxisomes counted. aZr1e59c8l0u.mpeLidvearn.dHtehpeatmoictyotcehoenxdhriibaitsshaopwoodirllayt-eddefcirinsetdaep.laRsEmRa misemdiblrataend.e. NTihnee (o9r)ganelles peroxisomes counted. Conclusions: Essentially normal liver. Poor cellular fixation. Average 5.4 peroxisomes per hepatocyte. 01573 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM: AnimalNo.: 195462 Treatment Group: 04mg/kg Sex: __non applicable. Species/Strain:RCart.CDOBRVAF/Plus Tissue: Lung Block No(s) 9_ 939.40 Z1591 Photo/Negative No(s): Z15985 Significant Lesions (check one): _____ Yes __X No Interpreting Pathologist (signature and date): ________ Features: ZI598L. Lung. Alveolus and lining cells. A partially detached pneumocyte is present in top margin. the air space. Large right The other dark organelles central appear cell contains a single lamellar body to be predominantly mitochondria. near The its cytoplasm glycogen. of these cuboidal pneumocytes contains a fine granular material suggestive of Z15982. Lung. Central cell contains multiple lamellar bodies and is the cytoplasm ofa ParNe2.at tThheisleifst oinfatnheaprheoatoofgrcaoplhl.apsTehdealtvweooldiaarnkdceslelpstao.nAeitchaeprilsliadreyofatnhdeenPdoNt2healriealnoctell cZl1e5a9r8l3y.ideLnutinfgi.ablTe.woTchoelbuomtntaormcrelilgshtocneltlheisraigPhNt1b.othNoconatbanionrmlaalmietlileasrdbeosdcireisb,eda.lthough the majority of their cytoplasm bottom of the photograph and a consist of capillary fine granular material. and endothelial cell are PN1s at the appear left. to be at the Z15984. round to pLoulnygg.onaAlncaelrlesaotfhatcoclolnatpasiendsaolmveeolliamaenldlasrepbtoad.ieAstctohnesitsotpenatnwdibtohtttyopme are IT two pneumocytes bottom PN2. (PN2s). Some lamellar material The central cells all appear to be appears to PNIs. At be the extracellular bottom right adjacent to the is a capillary, endothelial cell and erythrocyte. No lamellar bodies noted in alveoli. Z15985. Lung. several lamellar Ac the bodies, top and right is a sits next cuboidal PN2 lining the to a more flattened PN1. alveolus. This Centrally is a cell contains capillary containing two large endothelial cell nuclei and an erythrocyte. cCoonntcaliunsiinognlsa:melElsasrenbtoidailelsy wneorrmeaildefnettiaflieldu.ng.SoRmaenedxotmracteylpleulIlarpnlaemuemlolcarytmeaste(rPiaNl25) identified outside _one type II pneumocye. co1574 "TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 Animal No.: 19560-1 `Treatment Group: _1.6 mg/kg Sex: non applicable `Species/Strain: CiL.CDBR VAF/Plus Rat `Tissue: Liver Block No(s).: __99.39-21 Z15884 Photo/Negative No(s).:_Z15888 Significant Lesions (check one): X Yes No Interpreting Pathologist (signature and date): Features: Z15884. Liver. Hepatocyte. Plasma membranes and adjacent endothelial cells are indistinct, consistent with suboptimal fixation. Dilation of RER with some blebs. Some dilated cristae in mitochondria. A single clear vacuole is present in the cytoplasm on the right sideofthe cell. Fifteen (15) peroxisomes counted. 215885. Liver. Some granular deposits present in mitochondria and dilatation of RER. Twenty-one (21) peroxisomes counted. Z15886. Liver. Hepatocyte in center of photograph with hematopoietic cells at top. Some clumping of mitochondria and RER. Dilated RER. Six (6) peroxisomes counted. Z15887. Liver. Hepatocyte containing a single large lipid droplet in dorsal cytoplasm. At top right is a hematopoietic cell and at lower left is a bile canaliculus. There is clumpingofcytoplasmic organelles. RER and nuclear envelope appear dilated. Five (5) ppreersoexnitsoonmepshoctouongtreadp,h. Eleven (11) peroxisomes counted Z15888. Liver. Hepatocyte similar to those described above. Some stain precipitate is Conclusions: There appears (0 be increased numbers of peroxisomes per hepatocyte compared with control samples. Average 11.6 peroxisomes per hepatocyte. Suboptimal tissue fixation. 01574 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 AnimalNo:__19560-1 Treatment Group: _1.6 mg/kg Sex: _non applicable `Species/Strain:Ci.CDBRVAF/Plus Rat Tissue: Lung Block No(s).: __99.39-22 Z15889 - Photo/Negative No(s).:_Z15893 Significant Lesions (check one): Yes _X No Interpreting Pathologist (signature and date):-------- aaFrneedatpcuroronebtsaa:bilnysZeItxwSporHel.sasmeeLdulnlagam.ebolAdailrevsbe,ooduTishcesanlvdoeTwnrerihncogeulcgaehssohTecoompnealcolnaswwimolthpedrraomlkianrenynto vloaosnugpaertesy5 vwShPnoNl TFseehctricotno.aAlcicamplitlllaorayePiNsa.prneBseennetaatthheheboPtNoTmils eexotfetnesipohontoofgraaphs.iNdoelnatmefllceaedlmaatetriofapllone of ZorIibSgl3ho9no0gf.chleLeuPnugsN. isaTherPePeNZnianwngidthcilcmloonratenaidlnaosccoa:Gscpcyetiompilalbalrsleme.amvieTdroleenaytc.saTpbiholdeaiceresyn,tarnAadtlHtdhiaenrtkgocpaellnewfdtiattnhhaen10otohne aZSrIaeSnaSdOtwLit.chheetLdoupnigin.gbhett,AwleAvecndoautrswk,oelnaitrnegneycadllPts asNhnpdriercnsaiepnniogtllitanhrcietehaseyvcCaotlp.ieoi.ThiensaPP2tN2csoornmteiwnyhsaetveral elnamdteoltalhaerrlbioumdmiieamrsewBpirtheaisnetnttseacyhnteopulpapsemr. ICeapalndarliocwesroingahitnonfghecpk hoegrryharpohc,yeNsoanldaimnaesr beyral b2be1en5te6wa5et2eh,ntthLweuonPgPN,2IFshesaltiaunrniensgg haaeelaslaveoimluusl.moutT,hihoesseolPdmeNsapercroovrmmeiidnenfnogZtIsReSvEEeRr9ala1nm.desAlasiunsmgoslueondPdsNe.d2Tbiyopsreoomesent r PZIRS8s93i.o n thLiusngp.hrotAonglrvaoplh.usTwhitehrcaspeivloelraarlycautbohiedabloprreerumcooeyncas,nTahlearmeealeernmoyticleaerlydisstuisncitieve itis Rr. No mel ters fs rset nh age spn Conclusions: Essentially normal lung. Several type Il pneumocytes with lamellarbodies were identified. Fixation was considered acceptable. c01578 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 Animal No: __195602 `Treatment Group: 6 mg/kg Sex: _nonapplicable Species/Strain: Cl.CDBR VAF/Plus Rat Tissue: Liver Block No(s). __00.39-23 Z15894 - Photo/Negative No(s)..__Z15898 Significant Lesions (check one): __X Yes ______ No Interpreting Pathologist (signature and date): Features: Z15894. Liver. Hepatocyte with surrounding hematopoietic cells. This cell appears partially detached from adjacent hepatocytes, although two hepatocytes are present in the lower right of photograph, all three sharing a common bile canaliculus. `There is clumping of mitochondria, RER and some peroxisomes. Some dilatation of cristae is noted. Thirteen (13) peroxisomes counted. Z15895. Liver. Hepatocyte appears similar to cell described in Z15894 with clumping of organelles. Some granular deposits noted in mitochondria. Fourteen (14) peroxisomes. N counted. Z15896. Liver. Hepatocyte has clumping of organelles with intervening cytoplasm filled dweintshefiannedgrcaonnutlaairn msaotmeeriaglr,anpurloabradbelpyospirtesdoamnidnadinltaltyedglcyricsotgaee.n. TeMnit(o1c0h)onpderrioaxiaspopmeeasr counted. 215897. Liver. Hepatocyte appears similar to those described above. Plasma `membranes are indistinct. A hematopoietic cell is at the bottom of the photograph. Fourteen (14) peroxisomes are counted. Z15898. Liver. Hepatocyte morphology essentially same as above. A bile canaliculus speerrvoexsisfooumresatctoaucnhteedd.hepatocytes in the top center of the photograph. Thirteen (13) Conclusions: There appear to be increased numbers of peroxisomes per hepatocyte. Average 12.8 per hepatocyte. Suboptimal fixation of tissue. co157 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 Animal No: _1__95602 Treatment Group: __1.6 mg/kg Sex: __non applicable Species/Strain: CiL.CDOBRVAFPlus Rar `Tissue: Lung Block No(s).: __99.39-24 715899 - Photo/Negative Nofs)..__Z15903 Significant Lesions (check one): __X Yes ____ No Interpreting Pathologist (signature and date): Fbaesaitluarrelsa:meZll1a5r89b9od.iesL.unAgd.jaAclevnetolluisniangndcellilnsianpgpceeallrs.toCbeentPrNa1lsc.ellAisfraeePNer2ytwhirtohcysteeveirsal present in a luminal the alveolar erythrocyte. space. At the left No abnormalities of the noted. photograph is a small capillary containing 2fi1v5e90l0am.ellLaurngb.odiAelsv.eoTlwuso alnadmellilnairngmyceelllisn.-lAitketsotprcuecnttuerersisina msiidngdllee PleNft2owfhpihcohtocognrtaapihnsare within capillary lumina. No lamellar material is identified within alveolar spaces Zl1y5mp9h0o1c.ytLeu.ngT.woAlcveenotleurslainnidnglicneilnlgs cbeolltsh. coNnutcalienaltaemdeclellalrfbroedeiiensaalnvdeoalruesPaNpp2esaresvetno be a ctohnotuagihnsonseomise"llaimghetl"laarnbdoodnieesisan"ddacrokn"n.ecLtasrgteo cthueboniedxatlfplantetuemneodcyPtNe1atselgomweenrtleft 2ch1a5r9a0c2t.erisLtuincg.ofAPN2csl.ustNeor olfampenleluamromcaytteersi,alfoisuridoefntwihfiiecdhwciotnhtianinallvaemoellalrasrpabcoedsi.es. Z15903, within a sLeputnagl.walAl.grTouwpoocfecllusbociodnatlaianncdleffltast,tbeunetdnponneeucmoonctyatiensdeafnidnivteinvterallamcealplialrlabroydies. No abnormalities noted. Conoencclluussiteornso:f PNT2y,pepoIslspinbelyumsoucggyetsetsivfeaiorlfyienacsrielayseiddenntuimfbieedrisnotfhiPsNa2n.imOatlh,earnwdisaet ulletarsatstructural morphology appeared essentially similar to control sections. 001578 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 Animal No. 9560-3 Treatment Group: _1.6 mg/kg Sex: __non applicable Species/Strain:RCartl.CDOBRVAFPlus Tissue: ___ Liver Block No(s).: 99.39.25 Photo/Negative No(s).Z:1_5Z9105490-8 Significant Lesions (check one): _X__ Yes No Interpreting Pathologist (signature and date): sFienautsuorieds:witZh15h9e0m4a.topLoiiveetr.icHceepllatooncyritgehtb.orBdielreecdanbaylihceuplaitaotcyttoepsaonnd tohnreleefts.idTeshearnedis clumping of larger organelles within the cytoplasm. cristae. Ten (10) peroxisomes counted. Mitochondria contain some dilated S2e1v5e9n05(.7)Lipveerr.oxiHseopmaetsoccyotuentweidt.h clumping of organelles. The RER appears dilated. Zis1c59l0u6m.pinLgiveorf.theHelpaartgeorcyotregawnieltlhesh.emaMtiotpoociheotnidcriceallcsonatlaoinng stoopm,eldeef,nsaenddebpootstiotms. anTdhere dilated cristae. degeneration. Fifteen (15) peroxisomes counted. Some peroxisomes exhibit marked 215907. Liver. Hepatocyte peroxisomes counted. morphology essentially same as described above. Seven (7) Zde1s5c9r0i8b.edLcievlelrs.wHietphactloucmyptienagdjoafcmeinttotcohcoenndtrriala vaenidn.REARp.peaTrwselsvimeil(a1r2)topeprreovxiiosuosmleys counted. iCnocnrcelausseidonnus:mbeCerlsluolfaprefrioxxatiisoonmseusbcoopmtipmaarledtowpiotohr.conTthreorlelaivpepre.arAsvetroabgeesl1i0g.h2tly peroxisomes per hepatocyte. 001578 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.:_418-013EM-9939 Animal No: __195603 Treatment Group: _L6 mg/kg Sex: _non applicable. Species/Strain: aCA.CDBR VAF/Plus Tissue: Lung Block No(s): __99.39.26 Z15910Photo/Negative No(s).._Z15914 Significant Lesions (check one): Yes __X__ No Interpreting Pathologist (signature and date): _____ Features: Z15910. Lung. Alveolus and lining cells. All pneumocytes are cuboidal in this photograph. The second cell from the right contains several lamellar bodies and is a PN2. The third cell also contains two small lamellar bodies. No other remarkable features noted. ZI5911. Lung. The dark cuboidal lining cell contains multiple cytoplasmic vacuoles and `aTphpeeaprrsetuomboecatePNat2taolptrhioguhgthaldsefoicnointtivaeinlsam1e-l2lavracsutorluectsurwehsicarhepnrootbawbitlhyianrteh/ewevraeculoalmeesl.lar bodies Z15912. Lung. Alveolus and lining cells. Along the alveolus pneumocytes all appear flaened and to be PN1s. At bottom of photograph in collapsed septal area is a cell containing multiple lamellar structures, cytoplasm ofaPN2. No lamellar structures are present in the alveolar space. PZ1N591l3in.esLtuhneg.remaAitntdoeprcoefnttehreaailrawragye PspNac2e.conAttaibnostmtuolmtirpiglhetlaamcealplialrlabroydiceosn.taiAnsd2arlkartghein white blood and nuclear cell. Dorsal envelope. to tis is an unidentified cell with dilated endoplasmic reticulum Z15914. Lung. Alveolar space and lining cells. Within the alveolus are multiple extracellular lamellar bodies. Lining pneumocytes in this region all appear to be PNIs. Several capillaries are present in the photograph. bCoodnicelsusidieonntisf:ied.EssSenotmiaelllyamneolrlmaarlbofedtiaelslwunegr.e SococmaesiotnyaplelIyI pprneesuemntociyntaclsveoalni.d lamellar 001530 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 AnimalNo: 195622 `Treatment Group: _1.6mghkg Sex: __non applicable. Species/Strain:aCL.CDBR VAF/Plus Tissue: _L __ iver Block Nofs).: __99.39-27 Z15915 - Photo/Negative No(s).:_ 215919 Significant Lesions (check one): __X__ Yes _____ No Interpreting Pathologist (signature and date): Features: Z15915, Liver. Hepatocyte and surrounding hematopoietic cells. Sinusoidal endothelium and plasma membranes are indistinct, but cytoplasmic organelles are fairly evenly dispersed. Dilated cristae are presenti the mitochondria. Nine (9) peroxisomes. counted. Z15916. Liver. Hepatocyte and surrounding hematopoietic cells. Dilated cristac are present in the mitochondria. Sinusoidal endothelium and plasma membranes are indistinct. Dilatation of RER. Eleven (11) peroxisomes counted. Z15917. Liver. Hepatocyte with clumping of organelles and washed-out appearance of much of the cytoplasm. Plasma membrane i indistinct. Twelve (12) peroxisomes counted Z15918. Liver. Hepatocyte morphology similar to those described above. Mitochondria contain some dense deposits and dilated cristae. Eight (8) peroxisomes counted. 215919. Liver. Hepatocyte contains multiple clear vacuoles and marked dilation of RER. Mitochondria contain some dense deposits and dilated cristae. Thirteen (13) peroxisomes counted. Conclusions: Poor cellular fixation of iver tissue. There appears to be a slight increase. in the number of peroxisomes per hepatocyte. Average 10.6 peroxisomes per hepatocyte co1s58 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-09.39 Animal No: _1_ 95622 Treatment Group:_L.6 mg/kg Sex: __non applicable `Species/Strain: CAL.CDOBRVAFPlus. Rat Tissue: Lung Block No(s): 99.3928 Z15920Photo/Negative No(s).._215924 Significant Lesions (check one): _____ Yes ___X_ No Interpreting Pathologist (signature and date): wFehaitcuhrceosn:taZiIn5s9t2w0o,dLiusntign.ctAvlavceuoolleuss,aannddlaipmipnegarcesll(s0. beAt PceNn2t.er Trihgehsteisapapreoaurncdoendsilsitneinntg cweiltlh "empty" lamellar body vacuoles. The lining cells on either side of this round cell are flattened PNIs. Similarly, at lef central is a cuboidal cel protruding into the lumen which contains two pvnaceuuomloecsywtheicwihtahopuptearreemdartokabbeleemdpisttyinlgaumieslhlianrgbfoedaiteusr.e,Tahned cisolpurmobnaabrlcyelalPatNthewhliocwherhalsefnt'tis a 7[1592r1. eLung. Alveolus and collapsed septa. At bottom right is an oval cell whose cytoplasm contains one round vacuole and multiple open clefts that appear (0 be "empty" lamellar bodies. "The dark oblong nucleus to is op left is an endothelial cllofa capillary containing a dark erythrocyte. Lining the septum on (0p of this is fattened PN1. Another capillary isi the 7ce1n5t9e2r2o.f tLhuengc.ollAalpvseeodlaurseaa.nd lining cells. The central dark lining cell contains many prominent j`umsittotcohtohnedrlieaf,t obuftthailscsoelclo.ntAatintahsefteowp visacaunoelnedsotwhheilcihalmcaeyllbneucllaemuesl.lar bodies. A larger PN is Za1p5p9e2a3r,s toLubnege.xtrIanctehlleucleanrtlyerwoifthtihneaphcoaptiolglraarpyhspiacae.scroDlolresdalllaymealnldarveonrtmryaelllyina-rleikceapfiilglaurrieetshat c2o1n5t9a2i4n,ingAlevreyotlhuroscayntedslainnidnga cleyllmsp.hoAcystieng(lveendtarraklltyy).peTIopintseruimgohctyitsealiisnpirnegsePnNt,I.which contains emnudlottihpelleiallamceellllasrabnoddiceosn.taIitniisngboeurnytdherdocoynteesitahreerpsriedseebnty aMtatttopenreidghPtNaTn.d lCoawpeirlllaerf.iesNloineldambeyllar bodies evident within alveolar spaces. Conclusions: Essentially normal fetal lung. Some random type Il pneumocytes containing lamellar bodies were identified. No lamellar bodies were seen in alveolar spaces. 00155). `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 Animal No: __15962:3 Treatment Group: _L.6 mg/kg Sex: __non applicable. Species/Strain: CL.CDBRVAE/Plus Rat Tissue: Liver Block No(s).: __99.39-20 715925 - Photo/Negative No(s).:__Z15929. Significant Lesions (check one): X_ Yes No Interpreting Pathologist (signature and date): : Features: Z15925. hematopoietic cell on Liver. Hepatocyte the right. There is surrounded by other hepatocytes and a clumping of the cytoplasmic organelles and ldeanrsgietaireesaasnodfswoamseheddiloauttedcycrtiosptlaea.sm.ThMeirteocishdoinldartiataioanppaenadrbtloehbabivnegsoofmReEgRr.anuTlwaernty-six (26) peroxisomes counted. 2h1e5m9a2to6p.oiLeitviecr.celHlespoantotchyetleefatpapnedarbsotstiommi.larTtwoehnetpya-ttohcryetee(i2n3)Z1p5e9r2o5x.isTohmeersecoaurneted. 215927. Liver. Hepatocyte with indistinct cytoplasmic borders. Thereisdilatation and blebbing of RER. Mitochondria appear to have some granular densities and some dilated c2r1i5s9t2ae8.. TwLievnetry.-oHneepa(t2o1c)ytpeeraopxpiesaormseesssceonutnitaeldl.y similator above cells. Fixation is suboptimal. Twelve (12) peroxisomes counted. 215929. Liver. Hepatocyte with indistinct cytoplasmic borders. There is dilatation of RER. Mitochondria appear to have some granular densities and some dilated cristae. Eighteen (18) peroxisomes counted. Conclusions: Increased numbers of peroxisomes per hepatocyte. Average 20.0 peroxisomes per hepatocyte. Poor, suboptimal fixation. 01583 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013 EM-99.39 Animal No: __19562-3 Treatment Group: _1.6 mg/kg Sex: __nonapplicable Species/Strain: C.CDBRVAE/Plus Rat Tissue: Lung Block No(s). 99.39-30 Z15930 - Photo/Negative No(s).:__Z15934 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): pFreeasteunr.es:OneZIPSN7230e.xhLiubintgs.psAeluvedoolpuasdinadoficyntgoplealssm. exTtoeondPinNgESntwoitthhelaamlveelollaurs.bodTiehs oarteher PprNe2secnotnitnatinhse saelvveeroallarlalrugmeenc.ytoCpalpailslmaircievsacwuiotlhese.ndoStohmeleiaslmcaeillstuabnudlaerrsyutrhfraocctyatnetsbaordeipersesareent ZbIe5n9e3a1t.h tLhunlgi.ninAglcveellosl.us and lining cels. Centally is dark PN2 which contains one distinct lapaemne-lflaacebdodnyucdloerussalbutto atlssonuccolnetuasi.nsThseevecrlall ilmammeedlilaatbeloydioenst3opPNo2f.thiTshdearokthcelrllicnoinntgaicneslsan appear to be PNIs. 215932. Lung. Alveolus and lining cells. Within the alveolusis a free and unattached cell b`uwthitchheiosnneotoncletahrelyloiwdeesntaifnidedmiadsdtloecerlilghttypaep.peaAlrls t0hecloinntianignspnseevuemroaclyvtaecsuoalpepseaarndtoobneefdliastttienncetd, allavmeeolllara lbuomdeyn,.ea PN2 which hs differentiated nto a PN. No lamellar bodies noted in nZuIc5l9e3u3s,shLouwngi.ngAclovnetoalinuss saenvderlailnivnagccueolllse.s,Twhheicmhidadplpeealriningbecl"lemwpittyh"onllaymeallsamralblodoiersi.oTnhoef aanngdulmaaryncuocnlteauisnsabocvouepilteseampPtNyTlnaumcelleluasr. vaTchuoelelsa.rgeThdoersoablloclnegarnupcnleeuumsobceynteeathhsthneotPNf2ltiesnaend ernedloetahseeldiallamceellllanusculrefuasc.tanStome small tubular srucures inthe lumen of th alveolus may be a2p1p5e9a3r4,flaLttuennge.d. NAloveloalmuesl,iairnbiondgiceesllssreanprdesceonltl.apsAedtahleveooplasrasptlaar.geTehnedoltihneilnigalprceelulmnouecyletussal and erythrocyte adjacent to it. No unusual findings noted. Conclusions: Several type II pneumocytes with lamellar bodies seen. Also, some lamellar substance within alveoli noted. 01554 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013; EM-99.39 Animal No.: __19568-2 Treatment Group: _3.2mg/kg, high-dose Sex: __non applicable Species/Strain:CELaCDBR VAF/Plus Tissue: Liver Block No(s).: __99.39-11 Z15799 - Z15802, Photo/Negative No(s).:_Z15909 Significant Lesions (check one): Yes X__ No Interpreting Pathologist (signature and date): ______ Features: Z15799. Liver. Hepatocyte with sinusoid containing hematopoietic cell at top and longitudinal segment of bile canaliculus at bottom right. Hepatocyte contains one distinct multivesicular body. Numerous mitochondria show some slight dilation of counted. cristae, and there is some dilated blebbing of RER. Sinusoidal endothelial cell at top is poorly defined, suggestive of suboptimal fixation. Five (5) peroxisomes counted. Z15800. Liver. Hepatocyte surrounded by hematopoietic cells. Six (6) peroxisomes Z15801. Liver. Hepatocyte surrounded by hematopoietic cells. Plasma membrane of the hepatocyte is indistinct and fixation is poor. . Mitochondria appear dense with some. granular deposits and dilated cristae. Dilated blebs of RER are noted. Three (3) peroxisomes counted. Z15802. Liver. Similar overall cellular morphology to Z15801. Plasma membrane is indistinct. . Mitochondria appear dense with some granular deposits and dilated cristae. Seven (7) peroxisomes counted. Z15909. Liver. Hepatocyte with granular cytoplasm intermixed with mitochondria, RER, and random peroxisomes.. Mitochondria appear dense with some granular deposits and dilated cristae. Nine (9) peroxisomes counted. 215798: Liver. No negative; no print developed. `Conclusions: Normal hepatocyte. Suboptimal fixation. Average 6 peroxisomes per hepatocyte. 00158 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013; EM-99.39 Animal No.: __19568-2 `Treatment Group: _3.2 mg/kg.high-dose Sex: __nonapplicable `Species/Strain:CRia:CDBRVAFRlus Tissue: Lung Block No(s).: __99.39-12_____ Z15803 - `Photo/Negative No(s).:_Z15807 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): ______ Features: Z15803. Lung. Air space lined by several cuboidal epithelial cells. These cells appear to be pneumocytes which have not flattened out into differentiated PN1s. They have fine granular cytoplasm without many distinct organelles. Dorsally are two. blood vessels lined by endothelial cells. No lamellar bodies noted. Z15804. Lung. Alveolus and lining cells. Two cuboidal pneumocytes are PN2s and contain lamellar bodies. Two adjacent PN1s are associated with these cells. Adjacent tissue is collapsed septa and cell types are less clearly definable. Z15805. Lung. A cluster of cuboidal pneumocytes similar to those described in Z15803. To the left is a capillary lined by an endothelial cell. Within the capillary are two lamellar-like structures that appear to be extracellular. No PN2s or alveolar lamellar `bodies noted. Z15806. Lung. Alveolus with lining cells. Pneumocytes are low cuboidal and appear to be PN 1s, although not completely flattened out. No PN2s noted. The cell to the right `with an oblong nucleus appears to be a fibroblast. It contains numerous RER and some collagen fibrils are present extracellulary. Z15807. Lung. Alveolus with two PN1s, although cells are not flattened. Adjacent tissue is collapsed septa and interstitium. No PN2s or alveolar lamellar bodies noted. Conclusions: Occasional type II pneumocytes noted. Some intravascular lamellar-like `material also present. Many type I pneumocytes have apparently not flattened out, probably normal fetal lung. 001588 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: 418-013;EM-0930 Animal No.: _19569-1 Treatment Group: 3.2 me/kg, high-dose Sex: __non applicable Species/Strain: RCaLt.CDBRVAFPlus Tissue: ___ Liver Block No(s).: __99.39:13 Photo/Negative No(s).2:1_5Z81548347- Significant Lesions (check one): __X Yes ____ No Interpreting Pathologist (signature and date): Features: Z15843. Liver. Hepatocyte adjacent tocentral vein. Abundant numbers of mitochondria and RER are present, but plasma membranes and sinusoidal endothelium are poorly delineated. Cristae are dilated and broken up in some mitochondria. Multivesicular body present. Eleven (11) peroxisomes counted. 215844, Liver. Hepatocyte with some rarefaction ofcytoplasm. Dilated RER are evident and some granular deposits are present in mitochondria. Eleven (11) peroxisomes are counted. 215845. Liver. Hepatocyte with hematopoietic cell in sinusoid at lower left of photograph. Mitochondria in hepatocyte contain dilated cristae. Two multivesicular bodies are present in the cytoplasm. The RER appears dilated with some blebbing. Cytosol appears rarefied and plasma membrane is poorly defined. peroxisomes counted. Eleven (11) 215846. Liver. Hepatocyte surrounded by hematopoietic cells. Morphology similatro those described above. A golgi associated with multivesicular body is present. The mitochondria contain multiple granular deposits and dilated cristae. Eleven (11) peroxisomes counted. ZL5847. Liver. Hepatocyte, similar to above descriptions. Plasma membrane is poorly defined and fixation is suboptimal. The mitochondria contain multiple granular deposits and dilated cristac. Nine (9) peroxisomes counted. Conclusions: Hepatocytes appear to contain slightly more peroxisomes. Average 10.6 peroxisomes per hepatocyte. Suboptimal fixation. No other difference noted compared with control. 0158% TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No. 418-013; EM:09.30 Animal No.: __19569Treatment Group: 3.2 mg/kg, high-dose Sex: _non applicable Species/Strain:ECAaLCDOBRVAFPUs Tissue: ___Lung Block No(s). __99.3Z91:514848 PhotofNegative Nos)..__Z15852 Significant Lesions (check one): ____ Yes _X No Interpreting Pathologist (signature and dues Fpreeasteunrte.s:ThZeIc3en8t4ra8l Lceulnlgh.asAtlvweooblaussilwairthvalciunoilnegscewlhlsi.chSepvreorbaalblcyubroeipdraelsepnrteluammoecllyatres are cbaopdiilelsa,ryalitshporuegshenttheatlalemfet.llar material is absent in them. A small portionof a septal c2e1l5l8a4p9p.earLsuntgo.coSnetcatiinotnwsohosmwasllsevvaecruaollceusb,ooindealwtiothcosloummenalripeipdi-tlhkeelimaaltceerlilasl,, Tpohsesicbelnytraal sleavmeerlallarlibpoiddyo.r Tlaometlhlearlovwaecruorliegsh;t othfisthciesllcealppiesaarscetlol bweitahmaancriorrpehgauglear. nuAclceaupsilalnardy is in tZhIe38c5e0nt.erLoufngt.he pAhocltuosgtrerapohfwciutbhoiadnalenpdnoetvhemloicayltelisn,inwgitcehllmso.stly cytoplasm containing ofriniengtrhaenuallavreomlaatrersipaal.ce.NAoncihanrtaecrtseirtiaslticcelllacmoenltlaairnbsodaiseisngalree mpyreelsienn-tliinkethoersleapmreleluamrocboydtye,s bZIu58t5h1i.s ceLlulngis.notTwcloeaPrlNy2isdeanrteifpireedseansttoancdeltlhteyypec.ontain muldple lamellar bodies. A fZi1b5r8o5bl2a.stLaupnpge.arsAtcolucsotnetraionftphneeevlmooncgyatteesd onrucelpeituhseloialthceelrlisghatsodfesthcersiebePdN2isn.above pahpopteoagrr(a0phbse.pnTehuemotoepytceelsl (cPonNt1asi)nswhaiccohuphlaevelna'mtelfllaartebnoeddieosuta,nodriPs Na2PsN2i.n tTrahnesicteilolnsto PN Cnounmcelruosuisoncsu:boiEdsaslenptniaelulmyoncoyrtmeaslwlhiingc.h aSpopmeaerrtaonndootmhaPvNeZSflwaetreneeiddoenuttified. There are 001583 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No: 418-013; EM.99.30 Animal No: _19572:1 Treatment Group: _3.2 mg/ks, high-dose Sex: __nonapplicable `Species/Strain: Ci.COMBRVAFPlus fm Tissue: __ Liver Block No(s): __99.30-15 215853- Photo/Negative No(s).:__Z15857 Significant Lesions (check one): __X_ Yes No Interpreting Pathologist (signature and date): _______ Fdeetaatiulsreasr:e aZp1p5a8r5e3n.tlyLliovset.r. MPelmabsrmaanmeesmbarndancerisstaareewpiotohrilnymdietfoicnheodnadnrdiamaarneyincdyisttoipnclta.smic Nine (9) peroxisomes counted. d2u15e8t5o4,pooLrivfeirx.atiSoinm.ilRarERto a2p1p5e8a5r3s,mpalraksemdalyandidlacteeldl.ulaTrhimretmebenra(n13e)spaerreopxoiosrolmyesdecfoiunnetded. ZheIpSa8t5o5c.yteLiavnerd.biHleepcaatnoaclyitceulwuistaht htehmeattoop.poiReEtiRc pcreolflsilteos tahreerdiiglhattemdaragnidnpalnadsmaanother membranes are 215856, Liver. indistinct. Twelve (12) peroxisomes counted. Hepatocellular morphology is similar to cells described above. Fourteen (14) peroxisomes counted. 2m1e5m8b5r7a.neLsivaerre. poHoerplaytdoecfyitneeda.ndEsiugrhrtoeuenndi(1n8g)hpeemraotxoiposioemteiscccoeulnlst.edP.lasma and cellular wCiotnhcilnushieopnatso:cytHeespaatsoccoymtpesarpeodorwliytfhicxoedn.troTlhse.reAvaererahgieghe1r3.n2upmebreorxsisofompeesropxeirsomes hepatocyte. 0155 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013; EM-99.39 Animal No.: __19572Treatment Group: _3.2mg/kg,high-dose Sex: __non applicable. Species/Strain: CrL.CDOBR VAF/Plus Rar ES Block No(s). __99.39-16 Z15858 Photo/Negative No(s).:_Z15863 Significant Lesions (check one): Yes Interpreting Pathologist (signatwre and date): __X_ No Flaemaetlulraersb:odZy15to85t8he,iLguhntgo.fiAtslvneuoclluesusa,nadPlNi2n.ing Tceolls.its Cleefnttirsala icanpiinlglacreyllcoanptpaeianrisngtoacnontain one esruybtohprtoicmyatle fainxadtiloinn.ingSeonmdeothceellliuallarcedlelbriPslaissmpar.easnendtcienlltuhlearalmveemolbarrasnpeasce.are poorly defined due to Zb1o5t8t5o0m.rigLhutngb.othAclovnetoaliunssaonmdeplraemdeolmlianrabnotdlyy cvuabcouiodlaels lainndinagpcpeellasr. toLibneiPnNg2cse.ll oTnheriogthhtearntdwo 2p1n5e8u6m0o.cytLeusnga.ppCeoalrlatopsbeedPsNespt.umNaonsdpelciinfiincg caebllnso.rmaMloisttieasppnoetaerd.to be type I pneumocytes, although Z15861. not flattened out. Lung. Large top cNeontlearmceelllarisbaodpineesumiodcenyttiefi.ed.AtFtixsadtoirosnalis mpaorogri.n is vacuole that may bsiegahnt i"seamspmtayl"l lpaomretliloanrofbodayc.ellOtwhietrhlisnoinmgecellylssosaopmpeesartthoatbaeppteyaper tIopcnoenutmaoicnytseosm.e cArtysbtoaltstoomr pnousmseirbolyuslacmyetlolpalradsembircisi.ncTlhuissiocnesllismaagyrabneualoPcNyt2eowritahmiancarocpahpailglea.ry,Tihtseecnedloltaheltioaplrniughctlewuisth directly below the granulocyte. ZliIg5ht8.62T,heLucnogl.lapCsoeldlaarpesaedcosnetpatian.s Lsienveirnaglthliisgahrt-essteavineirnaglpfnleauttmeonceydtpenseuamnodceyntdeost,hebloitahl cdealrlsk.anAd. gbrealnounlgo(c0ytaefiisbrporbelassetn.t iNnothaebcnaoprimlallairtyiaetstnohterdi.ght.NoThlaemeolvlaalrnbuocdlieeusscnleexatrloy etvhiidsevnetssienlthaipspears to 21ph5o8to6g3r,apLhu.ng. Collapsed septal rea. The lamellar myelin-like material in the top center is wmiatyhibneacpraepsielnltariyn ltuhmeeanl.veoTlhuse. topAtaitrhsepbaoctetiosmlilenfetdibsya adafrlkaettrecneeldl PwNhIi.chScoomnteailnasmealllaarmemlaltaerrial bod `Conclusions: preumocytes. Essentially normal fetal lang. Fixation was suboptimal. Some lamellar bodies noted in type IT 001599 `TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _418-013; EM-99.39 `Animal No.: 195774 "Treatment Group:3.2mg/kg,high-dose Sex: non applicable `Species/Strain:Cr.CDBRVAF/Plus Rat `Tissue: Liver Block No(s).: 99.39-17 Z15864 - Z15868 Photo/Negative No(s).: __Z15940 Significant Lesions (check one): __X Yes Interpreting Pathologist (signature and date): ______ No FPelaatsumraesa:nd Zce1l5l8u6la4r.meLimvberr.anHeespaarteociyntdiestwiintcht sduurerotounpdoionrg feirxyatthiorno.cytMeistoicnhsoinndursioiadss.how some granular deposits and dilated cristae. Three (3) peroxisomes counted. ZL5865. Liver. Hepatocyte surrounded by adjacent hepatocytes and hematopoietic cells. A bile canaliculus is present at the bottom of the photograph. Eleven (11) peroxisomes counted. Z15866. No negative. ZI5867. Liver. The plasma membrane is indistinct. Eleven (11) peroxisomes counted. ZpR1eE5rR5o.6x8i.PslomaLesismveasr.hmoewHmebaprdaavtnaoecnsycateredeadneignddeiansdetjrianaccttei.notn.Shiexmtaeteonpo(i1e6)tipcercoelxlsi.soDmielsatcioounntaendd.blSeobmbiengofotfhe Z15940. Liver. Hepatocyte with poorly-defined plasma membranes. Mitochondria show dilatation of cristae. Fifteen (15) peroxisomes counted. Conclusions: Hepatocytes exhibit features of poor fixation. There appears (0 be a slight ihnecpraetaosceytien.the numberofperoxisomes per hepatocyte. Average 11.2 peroxisomes per 001599 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013; EM-99.39 Animal No.: __19577-4 Treatment Group: 3.2 mg/kg, high-dose Sex: __non applicable. Species/Sirain: CL.CDBRVAF/Plus Ra Tissue: __ Lung Block No(s).: __99.39:18 Z15869 Photo/Negative No(s).:_Z15873 Significant Lesions (check one): _____ Yes Interpreting Pathologist (signature and date): _X__ No rFiegahttucroenst:ai1ns58s6ev9e,ralLluanmge.llarAlbvoedoileasrasnpdacies aanPdN2l.iniAngdceeltlas.cheCdencterllail pjnuesutmaobcoyvteetohins one oabnldomngayenbdeotaheplniealumcoelclytnuecloerupso.ssibly a lymphocyte. Al the top left is a capillary with an Zse1v5e8r7a0l.cyLtuopnlga.smAircevaaocfuocloelsl,aptsweodoaflvwehoilcarhsceopntat.ainCelnatmreallllayribsoadydamraktecreilallt.haTt wcoontcaeillns bwietnheaatbhutnhdiasnctelmloarlesoclceoanrtaciyntospimlialsamracpypteoaprlatsomibce tvyapceuoIlpenseaunmdocmyatyesb,eaPlNth2osu.ghOtthheeyracreells not flattened. ZIS871. Lung. and covered by Ventrally it contains a PN lines the airway space. multiple smal clear vacuoles, The but smaller lamellar dark cell material adjacent is not to ccellelasr,lycaepviildleanrty.luDmoerns,alplnyetuhmeoctiystseuse,coanntdaiinnstecrostliltaipusme.d Nseoptaabnwoirtmhalaimtiixestunroeteodf. endothelial ZL5872. vacuoles Lung. Area and lamellar of collapsed alveoli bodies. Ventrally is and septa. a capillary Centrally are containing a two PN2s containing lamellar-type body within its lumen. `2T1h5e87ce3n.traLlucnegl.l cVoennttarianlslya caoreupsleevesrmaallclulbaomiedlallarp-nteyupmeosctyrtucetsurleisniinngiatsncayltvoepollaasrms.pacTeo. the sight and the top is top right are capillaries containing a flattened pneumocyte. some lamellar material in their lumina. At sCtornuccltuurseisonpsre:senNtowrimtahlinlucnagp.illSaorymelumtiynpae.Il pneumocytes identified. Some lamellar co159%, TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 AnimalNo: 195793 Treatment Group: _3.2 mg/kg, high-dose Sex: _non applicable Species/Strain:CRaHsL.CDBER VAEPIs `Tissue: Liver Block No(s): __99.30:19 Photo/Negative No(s).2:1_5Z81754878 Significant Lesions (check one): __X_ Yes No Interpreting Pathologist (signature and date): _______ aFnedatsuirneuss:oiZdIc5o8n7t4a.iniLnigveerr.ytHheropcayttoecsytate uspapmeorurnidghetd. by En d`ohtehmealtioaploiceetlil ci sceplol oartlylodweelri nleeaftt e d and hepatocellular counted. plasma membranes are indistinct. Seventeen (17) peroxisomes Z`b1o5u8n7d5a,rieLsivaerr.e iHnceopmaptloectyeteonmitxhiesdcwelilt,hasnodmpeerhoexmiastoompoeiseatriecncoeltlsc.ouHnetpeadtofocreltlhuislacrell. eGnrdaonpullaarsmdiecporseittiscualruemp.resent in some mitochondria. There appears to be abundant rough Zmi1t5o8c7h6o.ndrLiivae,r.endHoepplaatoscmyitcerewtiitchuliunmdi,stainndctpperloaxsimsaommeesm.braTnheirst,y b(u3t0)copnetraoixnissoambeusn.dant counted. 2S1o5m8e77g,ranLuilvaerr.depHoespiattsocayretep,ressiemnitlawritinhionvesroalmlemmoirtpohcoholnodgryiat.o tEhnodseopdleasscmriibcedreatbiocvuel.um ZaInSd8n7u8c.leaLriveern.velHoeppeataopcpyetaer,disliamtielda.r iTnwoevnetrayl-lfomuorrp(h24o)lopgeyrotxoitshoomseesdceosucnrtiebde.d above. Sanodmenugcrlaenaurleanrvdeelpoopseitasppareearprdeislaetnetd.witShiixnteseonm(e16m)itpoecrhooxnidsroiam.es Econudnotpelda.smic reticulum Capopnecalrussitoonbse:inTcirsesauseedfinxuatmiboenrssuboofpptiemraolx,isboumtebsetpteerrhtehpaantoscoytmee. prAevveiroaugsean2i1m.a7l5s.peTrhere hepatocyte (four cells counted). Noother changes noted compared with control livers, 01593 TRANSMISSION ELECTRON MICROGRAPHINTERPRETATION Study No.: 418-013 EM-99.39 Animal No.: _19579-3 `Treatment Group: _3.2 mg/kg, high-dose Sex: non applicable Species/Strain: CiCDBR VAF/Plus Rat `Tissue: Lung Block No(s). 99.39-20 Z15879 - Photo/Negative No(s).:__Z15883 Significant Lesions (check one): Yes X No Interpreting Pathologist (signature and date): Fc1e0elabltecuorinnetstah:insZaIlsSve8evTeo9rl.aal llLuaummneeglnl.,arbAbulotvdneiooelslsasa,mnendlilisanagmaPctNeo2rl.isaal.SnodTmcoeolhmleiaspicsgeehdltlaalinvee3ooulcsaercleslceloupnlataaraidrneeibnargiTssheeavpcTpeaeilanrgs cmyatcorpolpahsamgiec.vaOctuhoelresceallnsdaappeiarregtuolbaer ncuocmlperuess.seTdhipsrmeaaymobcey3esPN2, but also may bes arZupIpp5te8ua8rr0es,dtL(obuanregi.feaxctCtreaancltellryla?ul)llayBir,Nsa.ltfiOhnoteuhgeghrraictneualllpsaprelaimnraistnegmriitaxhleedcaoiwnritwtaahiynoiatnphgpeermaucrletltilpllbaeelraPdmeNbeTrli.lsaabnoddisesp.robTahbilsy a ZlmaaIrcSgreSopLvha,acguLeousln.egs.ActoAntrhaeeian(ioonpfgcaosnloellmalepispsetcidcaaanlltvvmeaaotcleiuroialaneld.aspeTppheiaears.seCmenactyornabtlealiyPnNar2seso,mseebvueltraamalellscloealmrlsamyawthebireicahl.conTthariene dorsal nuclei have dilated nuclear envelopes, suggesting suboptimal fixation. eZmla1tl5e8ri8in2a.tl haLpu"pnsegea.rpstTiuhnimctkahepepnaeeladvr,oscltoaolrlbalepusameePdnNs.oepntatlhTehweallelfit,gmhlitinxemedabdrygwiintphniesuiomnomceeydtcbeesyl.u3aPrSNodTem.beriTsf.hreeeTdlhaaermkecleslneatprraall ZerIulpSltEua8tr3et,dhePLNlu2ong.w.erAtrAigtrhoetpacioonfnttchaoeilnllsupmsseoenmdeaisllvmaeumoleltliilpaalrnedblosademipeetlasl.aarnSdbiom1diilepasr.rob1aTZbhl1eyc82o8lP0lN,a2p.tsheids smeipgihatarnegpresent a capillaries appear essentially normal Conclusions: Essentially normal lung. Some normal type Il pneumocytes identified. Some lamellarmaterial identified within alveoli. 01594 IIL Transmission Electron Micrographs 01594 NOTE: `The original glossy figure photographs (Figures 1 - 14) were delivered to Dr. Marvin T. Case with the draft pathology report on March 19, 1999. 001596 Figure Legends PriAmnecdiillcaarAyrPgautshoNloo.gy41R8e.p0o1r3t rMeapgonritffiicgautrieosna.re Partiantmamganginfiifciactaitoinonoifs1i1n,d2ic0a0tXe.d Tohnethmeebaascukroefmeenatchbaprhootnogcraacphh. All photomicrograph equals one micron (0.001mm). `EmiigtuorcehoL.ndrLiiavear.ndHreopuagthoceyntdeopslhaoswmiincgrneotrimcaullucmyrteofpllecatssmiscuboorpgtainmelallesf.ixaCtlioanm,piSnogmoef peroxisomes is present a are identified. the junction of Hematopoietic cells three hepatocytes in are the present top left. ventrally. A Animal No. bile canaliculus 1506-4, 0 `mg/kg/day (control), 11,200X magnification. iEnidgiusrtein2c.t pLlivaesrm.a Hmeepmabtroacnytee,.anDdiloartgatainoenlloefctlhuemrpoiungghiennddiocaptleassmuibocprteitmiaclulfuimx,atiaonn. Mitochondria representative are dense and peroxisomes show some are labeled. granular deposits and dilated cristae. An endothelial cell nucleus is present Some lining the s(cionnutsrooi)d,in11t,he20u0pXpemraglneiftficcoamtieorn.of the photograph. Animal No. 19509-1, 0 mg/kg/day lFaibgeulreed3a.ndLiavperp.eaCremllourlearnmuomreprhooulsotghyanisisnicmoinltarroltso.FiTghuerer2o.ugShoemnedoppelraosxmiiscomreestiacruelum aphnodtnougcrlaepahr eAnnviemlaolpeNoar.e1d9il5a3t4e-d1.,A0.4bimleg/ckagn/adliacyu,lu1s1,is2p0r0eXsemnatgniinfiucpapteironl.eft of the Einignuurmeb4e.r,Liavreer.labMelietdo.chSonodmreiahesmhaotwopdoiileattaitciocnelolfs carriestpaer.eseSntomien tpheeraodxjiascoemnets,siniunscorieda.sed Animal No. 19560-2, 1.6 mg/kg/day, 11,200X magnification. wFiitghuirne m5.itoLcihveorn.driHae.patSoevceyrtaelwpietrhoaxnisiondmiesstianrcet lpalbaeslemda.meAmnbirmaanle.No.Cr9i5st7a7e-a4r,e3d.i2lated mg/kg/day (high-dose), 11,200 magnification. Figure 6. Liver. peroxisomes are lHaebeplaetdo.cyAtneiwmhaolsNeo.pla19s5m7a9-m3e,m3b.r2amnge/kisg/inddaisyti(nhcit.gh-Rdeosper)e,se1nt1a,t2i0v0eX magnification. Fbiogduirees a7.reLpurnesge.ntA.lvTehoelucsenatnrdalldinairnkgcceelllls.maTywboe taytpyepIel Iplnpenuemuomcoyctyetsecownhtiacihnihnags lamellar expressed its lamellar bodies. top and bottom margin of the Type I pneumocytes air space. Subjacent can be alveoli seen lining the alveolus are collapsed and the at the Nroe.pre1s9e5n0t5ed-1c,el0lsmga/rkegp/ndeauymo(ccoynttreosl)a,nd11s,e2pt0a0lXcemlalgsniaftidciaftfieorne.nt functionsof cut. Animal 0159 PrimAendciilclacAyrgPautshNool.ogy41R8e.p0o1r3t Eigure 8. alveolus. TLhuinsg.celAllvceoontlaiiansndsesveeprtaulml.aAmeltlayrpeboITdiperseucomnotcayitneinigs present lining the right faint laminated material. pTrheeseonttheinr ltihneinsgepcetlulms aatrethteypteopIporfetuhmeopchyotteosg.rapAhn. eAndnoitmhaelliaNlo.cel1l89a1n5d-1ca,p0ilmlagr/ykgar/eday (control), 11,200X magnification. Figure9. Lung. septa containing cAapilllvarieesaonwidltsheupltsuam.inTalheercyetnhtrroaclytneasr.roAw alveolus lamellar is bounded myelin-like by two structure is aplrveesoelnuts.in Nthoe tlyupmeeInI pofnetuhemoucpypteerscaoprilllaamreyl.laFrlabtotdeineesdatryepeeviI dpennetuminocthyitsepsholtinoegrtaheph. Animal No. 19504-3,0 mg/kg/day (control), 11,200X magnification. lFaimgeulrlear10b.odLiuensgi.s pArlevseeonltucsenatnradlllyi.ninMguccelhls.ofAthetylpaemeIIllpanremuamtoecriyatlehcaosntwaaisnhiendg-mouulttiopfltehe slianmgellelalrambeodlileasr.boAdy.secTohned rteympaeiInipnngecuelmloscayptpeeaisr ptroebseenttynpeeaIrptnheeubmootctyotme,sa.ndAnciomntaalinNso.a 19546-1, 0.4 mg/kg/day, 11,200 magnification. cFoingtuarieni1n1.g sLeuvnegr.al Allavmeelollaursbaodnidesl.iniTnhgecelalds.jacAetntthleintionpg ciselalscuabpopiedaarlttoybpee pITnpenuemoucmyotceyste which have not capillary spaces. flattened out. Animal No. The lamellar myelin-like 19560-2, 1.6 mg/kg/day, structures are extracellular 11,200X magnification. within lFiikgeurmaeter12i. alL,unwgh.ichAlivsecoolnussisatnendtlwiniitnhgscuerllfsa.ctaAnltvseeoclraertsepdafcreocmonttyapienIsf apbnuenudmaonctytleamse.llTarh-e rdaermkaicneillnuglalrinsienggmceelnlts naepapreatrhetobobtettoympemaI ypnbeeumaonceyxtteesnsoironcoufboa idtaylpepnIeIupmnoecuymtoecsy.te. The ACanpiimlalalriNeos. wi1t9h56l0i-n3i,ng1e.n6dmotgh/eklgi/adlacyel(lhsiagrhe-dporsees)e,nt11do,r2s0a0lXlymaagnndifatictahteiolno.wer right Figure lamellar 13. Lung. Alveolus bodies are present. and lining cells. Two A type I pneumocyte itsyppereIslenptncluinmioncgyttheesaclovnetoaliusnivnegntral to the type evident. II pneumocytes. The material in tWhietahlivneotlhuessiesptnaolndweaslclricpotllcaeglelnulfairbedrebsraisn.d a fibroblast Animal No. are 19569-4, 3.2 mg/kg/day (high-dose), 11,200X magnification. pFingeuurmeocy14t. esL.ungT.heAllavregoeliceantnrdalsecpeltlumi.n tBheotshepstiduemsaopfpethaerssetoptbuemaatreypleinIepdnbeyumfolcaytttee.ned T1w95o79ce3l,ls3.v2enmtrga/lklgy/cdoanyta(ihniglha-mdeolslea)r,bo1d1i,e2s00aXndmaagrneitfyipceatIiIonp.neumocytes. Animal No. 001593 Abbreviation A BC BE cL Cap oN co cp D DD DC DN DR EC ER FB G HC L LB M N Pp PNI PN2 PM RER PriAmnecdiilclaarAyrPgaotshoNloo.gy41R5e.p0o1r3t Index of Labels forUltrastructural Structures Structure: Alveolar space Bile canaliculus Bronchiolar epithelium Capillary lumen Capillary Centrioles Collagen Cuboidal pneumocyte Desmosome Dense deposits Dilated cristae Dilated nuclear envelope Dilated RER Endothelial cell Erythrocyte Fibroblast Glycogen Hematopoietic cells Lipid droplet Lamellar body Mitochondria Nucleus Peroxisome Type I preumocyte Type Il preumocyte Plasma membrane Rough endoplasmic reticulum 001594 o"l3Me 1 3 SE me ST i Cog Se A Ree R paiHdE E eeR A(SUS DAEyRa85 Se MaBYdEjdTimAiSE EiAE ne sEapinleEe' 3 A ERT bees ey WE SAFER ien AR ER TE LF I 8 rate ol ale er - ey Yon strat wid CH 557 Ne Tea iy aNy Fonsi oy Re Coen Lee EEE paid DoUurASm EEEl Rpe A lSE TETA SlRE F7a CdAoAlE lrin] . as A Nho. F0 SC SoEoEhrH Sizes frosn LEeRt & jjorTologe - p le e Fa 3 P TERNy AL TET pAr kg =. LAE TE E ATR Kipp ERs Ch RRA Tad A nals TE HR Se EVE Eg SEARS 2 0Bs a. v RLakgiRoee "ea g SaE n - a A aAy r J. BheE ae ra s e n ams, 5 TR FF=3 o L(aeN SE GeH e e Paes g oo bL onE a E TR I T Aye F oN Te, g i can cae Le l e. e 7 Eo BEST COPY AVAILABLE 001600 Fr ns . yl X os. A BE hy Nod HE B DO= S a oe Ey: Ee SiTH o Seet LiTng, IE se FE a S308 . CT he E o EEL Cc SEE EET fn Ten SO SPRY SE A BY 2 : gt" FL Ee chXE Ck Rel NatsJ. OF Rg Lea a eg Pt 3 sR Ca RE 25) ' Xo Ev SR oF i ee 5 wi" opis BEST COPY AVAILABLE co160p BS a SR I RD ap WE CEE J Lot BAR ae GOEEE Saoloosh2heSdEO YOaRk,CWAaRnAS CGTf rrwaihAliS) CREBR ESFERT|CoS A AS CREE AER] Foie ld y beE r hl DER R il SNe eh Hee Ra IES U f 7 ofErAAATRFi at Se e Re C SECREGER Bo onl en Doreei AaRETO Ee eSCy Ry Sash ia ees Sate F vepRAiu lPaeNa ERoLak Sne aRa T TRd E li ael e S0T a hL o RR OiSOdONPIEC RSEOT e e a TEpiPEaSCEGR STAT Pa SeTlgl WRTgleNy ' PaEaL: E7 SuleN Nd eG, a BoleEamaSLe ea WTENEeR ater omIil SF NEE INT aIEL oRe SS ea T NENAes n Lh R WE ARe R 5 PI RN Fy 3 i meSEntt Sty 2 aA ET & oF . . % 3 LE$TSmTf =F oo. BEST COPY AV,AILABLE 001602 = soe i - 5 pee Wy he SY men IEAS, TM bee20 ag Lae 2 ay F A:a o RE a PS RA E AR a SREEdEdR B=HE iv ox & AoA NEST IES SEL, Ch Eee -- in By a Re, AEJeo RE fi ragEsl Se WEE GyFi oh RS 8 k elr loTA BNLOTYA pri 2 TARR 58 IEE ay ie :, BL TA ap Ee FAaTgE ene, & PS). VEE aw he. so GHEY gay Sh : WIR, E00 or IR Leg NEC TSONpE as EL BoHBC SN Lp : ChE pS ee * RRenO DSovisstsy Ao nnN i B Zalald or eR eg tnes Eh AE GDR x Sren nl STR ORNR RBE nT SEEREENEiaAnSseeT3] aN C EEN R S AE REelKU NAERN a dT EaEY b0 e R A R y E Sg we ow HES Se ea Soe BEST COPYAvan ar: 001603 En a : BbLLasr Ce N > < Hop: . A SE mee a a or CsU ERREE Eone TREerEeLe oy a JIB pa Y&ioLgose BSE ES ey SaAg SET VE At Tos EOE Na SORE of CE Wea AT Bare If % EY a Saks WY aN i 3 a? 5 BIC GUe AAPAN a a BENT ih PR Lo leah eT PERC oe B&ie( TIaire )LNe IEEEY EJthP y Lb ,ON Rd : Ak he REE SETA Se BR ial om els bh 0 SE Nga 3 = No on CWE E.bs |S Ge h TNPrRa , AER De ww blow BEST COPY AvAILARF oo1604 Baff WNL > DimWe CaF ns HE Lok) aR, Fo raia lTEBEe ans e ee Blan ee shSuo dOT E aR nRalday Vm T aA EE QRER URE rg BPELp a CRGT rede RTA DE e E i Bn in g ame a Co FDow iA riNF Tog NgTe ule esR oa pSS irTTL E ge aa Sr. a, BS las R vle AR Soe SlEnC dl E e ENeeyy e ER BEGT R EgE RL SaEsE T RTa ee wanNg T HA oie ga e 2I S E TI EN s CUTA IE AER e R AS EN F PoSent l etee eRa rees Ne E wi Boa 3 doeRakes aNIE Ti T NR jo S MS SAR TSEeg STT E sy TRE BESTCOPY AVAILABLE 001604 aE Need SL BRR J Nr RE LER RomeRA Se 3SotB s,oA fleg ETeALR ei EM RNlE ARa Ta h GY Wpar ge SIY RN ERE SE 1 I NY 5 R SReIB E g paS l SRE a NEaN eR AA aYpo iY g Sae Lid [S7 aNR ACheAR ULRGRE Sr ON REI y -@ LE Ls to A nT, edad . 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Kr Fuel XC Ni a RIESE SB ERoR RAHR Ld HiyAJLSRLEtET SR) ra wT i AY ol IS ROy= AGES RR ER RORERANICIR + SERRe So > Hy E 5 E, LR o SOWd PE. 7RR J enWaaaa O5ly8 va ior ta eToSEOER AToe BoogHgY A EREaREPeoroJebolg m Pe NG d oCAi I RT ENTE I No Lif a SO IPRoyESSPtas Saglge) REITre TardsR he H SR RI = AaSoc A CN ECAE RR A R apn r: SUaE] Sa SEIN EC ER C BRNNCEeLme A INeRe Bre oon) STA. SE Neal" lSeenSaasTi KE CERES g TEsT AflRaA t aD d HEL oF RC SA EER, inte AREA oS QpE aTheenyiAtE e Kiet EPS Sa HER sli B ao. i 3 =k i SAN of RES and DrABA Sel PCEO,R aA BSS B BORE E pov RwaS RIL TO BEST COPY AVAILABLE 0160 Bogor FE IN197) fr | EEE Wi SET mw oS BY EE CCIEhAi.Ce{Ce ARIN N oGga 9 t REAR en gan TB NEAAP, <a, Fenb =w (Rel SIRE ee Misa mF (30%LigTn tlE el . r 5 i Pie 47, LOSE ' "oA EE a 5 NST oY as ON CC 2 FIP eben ek H _ ls 3 BEST COPY AVAILARF co1660 N$ ENEn RE H Ah H RRe CisSEHE R ik d re EeesRE i e tlg fi, Nfy y Yo 4 0 oA N ThNAA d R yeuoNaaee Sg Th B ilS AL pi G fal & RAaSssEcptTI y A s a ly BEB (EA Na BeyV QUELTHheIMNees AY : yg |, OSTE RA S Te i ye w : CE mr ALE Ne To3h ~ aLRai g cp oy LT es | at oF e8a= h 5b 1 a 7 OE pe R gal 9 pon + =" $o2i he i he ns | JAea:d ~ Do LE B fm nine Wd Si" PT AYN E Ras R AW Lo>AYaL TEEc S SaT lbrekAgSasLEi LEdEE R1EER ET SEE he BESTCOPY AVAILABLE co161 VJECeWen E NO ATE P a a E ATRE ETnR ee Fh E T Soed i a ES CaeTHER TBen ED L7 aa S A dR TE SpeBr "ft =oF ABE g a To%ppihRgpo FaBiRtEE BA E 7 oh i ge BEE CE BR " 3 ae i BF Synf i TL Le EAH oil g oh RAE TRL yeu if RT Pi e - C. t aRRA ZooaWNEl 3 AN age geo ESE SANE NO ce eae R. | ET; Roan a FLO at pOtRhE ET SiEd 2)YeEll CONG EL - CST AT Rm 5 i FOU TE NEE ve a ---- at BEST COPYAvAM 52:r = TARASNerd 01612 WRN. Xi % qu ho 8 ACRE) CSRS le oo Se 70) Vis be | iy SE: oH8w a o SEE sRlRrEL`8 E (5Y LQ3 AaN-vAf RHea&jSon 3 Ba A NR 8 RNE T od aR stIX FNee fn AAARYEESe EE aX ce Si . NEE CMHEREEa E "d ELceie Pe | Ne SE L3 e AEeR MT Sa SRoEEEA E Py AL gPiTlhH EEEsS doNS lES eLsoe oEanEfeRefh GSEC aUlA S VTACRetd]A C aAN nLR NlI ElAegTT e S LJyN ly V STot AE RTTRI IBERRS L LE e e a e R P R nS R Fo ETBA as SONOR ed pGihBnt r eyR NShDelIE R A E BEST COPY AVAILABLE 001613 IV. Quality Assurance Statement co1614 ' 777/ PatCohmpoalnyoogfySlAesesAoppciicaatotnes stemIatnitonearinCoartpoiraotinoanl e =R=a .= QUALITY ASSURANCE STATEMENT AssuranTcheisUneiltecatsrornemquiicrreodscboyptyhestGuodyodhaLsabboereantoirnyspPercactteidcaensdReaguudliatteidobnsy tphreomQuulalgiattyed bQuyatlhietyUA.sS.suFroaondceanUnditDrwuhgicAhdmrienpiosrttrsatdiiornec.tlPyAtIo hmaasnaagfeumnecntti.oninTgheanfdolrleoswipnognsiisvae record of inspections/audits and their resulting reports: IDnastpeecattion PhaseInspected DtMaoatneFaignedimnegsnRteported andStudyDirector 2/22/99 022,23, 26,2589 2sio9 4/5/99 Processing and Embedding Data Audit Draft Report Audit Final Report Audit 2/22/99 2si99 esio0 4/5/99 C `Que aliety AdsesWuarradn,ceB.SSp.e,cRiaQlAisPt-GLP Date Sponsor: 3M Corporate Toxicology Test Article: PFOS Study Number: 418-013, PAI Study EM-98.39 01615 4915D Prospectus Drive Durham, North Carolina 27713 (919) 544-5257 (919) 544-3218 FAX