Document yrwo8myq2DEnMKB0zB69bB6j4

yn&uAt/uj&l BIO -TEST 1810 FRONTAGE ROAD NORTHBROOK, ILLINOIS 60062 9nc, REPORT TO MANUFACTURING CHEMISTS ASSOCIATION GROSS AND HISTOPATHOLOGIC FINDINGS ONE-YEAR INTERIM SACRIFICE CHRONIC VAPOR INHALATION STUDY WITH VINYL CHLORIDE (ETHYLENE DERIVED) IN ALBINO RATS MAY 9, 1975' IBT NO. 663-03222 CAM 102254 %AuMmclL BIO -TEST JlaIxybGJ/yu&, Dnc. 1810 FRONTAGE ROAD NORTH BROOK, I LLI NOIS 60062 May 9, 1975 Mr. Milton Freifield Project Manager, Vinyl Chloride Research Manufacturing Chemists Association 1825 Connecticut Avenue, N.W. Washington, D. C, 20009 Dear Mr. Freifield: Re: IBT No. 663-03222 - Chronic Vapor Inhalation Study with Vinyl Chloride (Ethylene Derived) in Albino Rats - Gross and Histopathologic Findings - One-Year Interim Sacrifice We are submitting herewith our laboratory report prepared in connection with the above study. Very truly yours, JCC:_mcb J. C. Calandra President CMA102255 JtvL^uaL B I 0 - T E $ T 2woMii&ML, REPORT TO MANUFACTURING CHEMISTS ASSOCIATION GROSS AND HISTOPATHOLOGIC FINDINGS ONE-YEAR INTERIM SACRIFICE CHRONIC VAPOR INHALATION STUDY WITH VINYL CHLORIDE (ETHYLENE DERIVED) IN ALBINO RATS MAY 9, 1975 IBT NO. 663-03222 I. Introduction At the request of the Manufacturing Chemists Association, 40 rats (5 males and 5 females) from the control and test groups were sacrificed on September 5, 1974, to conduct cytogenetic studies on bone marrow specimen obtained at time of sacrifice. The purpose of this report is to present the gross and histopathological findings in other tissues from these animals. CMA 102256 ^nduUMai B 1 O - T E S T 2inyiai&tid.. II, Summary Treatment-related gross and/or histopathological findings were confined to the liver of test animals at exposure levels of 50, 200 and 2,500 ppm. Gross pathologic findings ascribed to vinyl chloride were confined to the liver of 2 animals of the 200 ppm level (group T-II) and the liver of 5 animals of the 2,500 ppm level (group T-III) . Findings consisted of solitary, dark red, nodular foci in the subcapsular region of single liver lobes. These nodules varied in size from approximately 4 mm to 4 cm in diameter and, on cross-section, they contained blood-filled cystic spaces. Upon histopathological examination, the above lesions were identified as sites of hepatic angiosarcoma. In addition, there was focal hypertrophy and hyperplasia of hepatocytes in the liver sections from test animals at each exposure level. The incidence and relative severity of this finding could be correlated with the level of exposure to the test material. No extra-hepadc angiosarcomas were observed. Materials and Methods All animals, selected at random for sacrifice, were rendered unconscious in a closed chamber by exposure to carbon dioxide gas and exsanguinated, and a complet*- necropsy examination was performed. The organs were weighed. Representative specimens of the following organs and tissues were placed in 10% neutral buffered formalin for fixation: CMA102257 JrtdndCUcU B I 0 * T E 5 T J,a&yuzbyuea, ,f*e. 3 Adrenal Glands All Gross Lesions Bone (femur and sternum) Bone Marrow (sternal) Brain and Spinal Cord Esophagus Eye Gonads (testes and ovaries) Kidneys Large Intestine (caecum and colon) Liver Lungs Lymph Nodes (tracheobronchial, cervical and mesenteric) Optic Nerve Pancreas Parathyroid Gland Pituitary Gland Prostate Gland Salivary Gland Seminal Vesicles Skin Small Intestine (duodenum, jejunum and ileum) Spleen Stomach Thymus Thyroid Gland Trachea Urinary Bladder Uterus The above tissues from all animals were processed by conventional methods, embedded in Paraplast*, sectioned at 4-6 microns, stained with Hematoxylin and Eosin and examined by light microscopy. The gross pathologic findings are presented in Tables I through II. The histopathological findings are presented in Tables HI and IV, The pathologist's report is presented on pages 4 through 6. Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. Report prepared by: Donovan E. Gordon, D.V.M., Ph.D. Section Head, Pathology Report approved by: Manager, Toxicology mcb * Sherwood Medical Industries, St. Louis, Missouri 63103. CMA 102258 jruLtdi'ual B I O * T 5 T 2MMyxtf&ue4.. J*c. -. I3T No. 663-03222 Manufacturing Chemists Association February 27, 1975 A gross and histopathological examination was conducted on a series of tissues and organs from rats of the 12-month interim sacrifice of IBT No. 663-03222. Treatment-related gross pathologic changes were confined to the liver of 2 T-II females (Nos. 550 and 5 99), 1 T-III male (No. 694) and 4 T-III females (Nos. 717, 732, 735 and 742). The lesions consisted of solitary, dark red nodules within the subcapsular region of 1 liver lobe. The nodules measured approximately 4 mm to 4 cm in diameter and contained blood-filled cystic spaces. There were fibrous adhesions between the capsular surface of some of these nodules and adjacent visceral structures. In 1 animal (No. 735), there were 2 different liver lobes with solitary nodular foci similar to the above. Upon histopathologic examination of the hepatic nodules, they were found to be angiosarcomas. A small amount of hemorrhagic fluid was present in the abdominal cavity of 1 animal (No. 59 ) as a result of hemorrhage from the surface of the liver tumor. Treatment-related histopathologic findings were confined to the liver of 5 T-I animals and all of the T-II and T-III animals. There was an exposurerelated incidence and severity of hypertrophy and hyperplasia of hepatocytes throughout the liver sections. The hypertrophied hepatocytes had an increase in cytoplasmic mass which had a light, eosinophilic staining appearance with a few basophilic condensations. In some sections, these cells formed small CMA102259 jndt*&t'uaL 8 I O - T E S T J&ua'ia&u&i, Jhc. 3 nodules with very little compression of the adjacent parenchymal cells of the liver. Some of the hyperplastic hepatocytes had prominent nuclei and nucleoli. They also formed small, discrete nodules in some areas. In addition, these cells often had a slightly increased cytoplasmic basophilia. In regard to the angiosarcomas, the early and more advanced stages of the tumor could be identified. In the early lesion, there was focal sinusoidal dilatation accompanied by hyperplastic and/or hypertrophied endotheliallike lining cells . The areas usually communicated with frank areas of angiosarcoma. The latter were characterized by the presence of blood-filled cystic sinusoidal spaces (peliotic foci) in which there were papillary pro jections of cords of liver cells that were covered by large, hyperchromatic to pleomorphic lining cells on the sinusoidal surface. The hepatocytes in the area adjacent to the tumor were also invaded by these tumor cells. In some areas, there was complete destruction of the enveloped liver cords and replacement by solid sheets of sarcomatous tumor cells which contained numerous mitotic figures and had anaplastic features. In most instances, the tumors were situated just beneath the capsular surface of the affected liver lobule and there was focal invasion of tumor cells into the capsule with hemorrhage into the peritoneal cavity at these sites. There were also fibrous adhesions to the adjacent visceral organs at the capsular surface of the tumors which correlated with the gross observations. In 2 animals (Nos. 663 and 668), the tumor foci were too small to be detected microscopically and they were in the early stages, of development. In this study, there were no metastatic CAM 10226Q 'ridutU'Ual 3 I O - T E S T jiauiyioAyueA., J*ic, tumor foci found in other organs , however; tumor emboli were present in some of the vessels of the larger liver tumors. All liver lesions, observed by light microscopy, summarized in Table III. The other changes described in the liver and other tissues (Table IV) were regarded as lesions of naturally occurring diseases. The lesions were present, in most instances, among both control and test animals. The small (1-3 mm), red, subcapsular foci observed at necropsy in the liver of both control and test animals could be correlated with scattered foci of hepatocellular necrosis observed histologically in these animals. Donovan E. Gordon, D.V.M., Ph.D. Diplomate, American College of Veterinary Pathologists CMa 102261 jndudi'ual B I O * T E 5 T JzitO'iaiyued, TABLE I TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Individual Gross Pathologic Findings in the Liver - Males Group Animal Number Gross Pathologic Findings Control 39 54 55 69 84 1 mm dark red foci in all lobes None Few 1 mm dark red foci in all lobes Numerous 1 mm dark red foci in all lobes None T-I 205 256 264 279 299 Few 1 mm dark red foci in all lobes 1-3 mm dark red foci in all lobes None None Numerous 1-3 dark red depressed foci in all lobes T-II 404 433 459 469 489 Numerous 1-5 mm dark red depressed foci in all lobes Few 1-3 mm dark red foci in all lobes Few 1 mm dark red foci in all lobes 1 mm dark red foci in all lobes 1 mm dark red foci in all lobes, slight pale discoloration T-III * 609 639 663 668 694 None Numerous 1 mm dark red foci in all lobes Granular surface with numerous 1 mm dark red foci in all lobes 1 mm dark red focus in 1 lobe 0.5 cm dark red subcapsular nodule in 1 lobe with capsular adhesions to spleen CMA 102262 yndwibiial B I O * T E S T Jlalttyiato/u&i, Jnc. TABLE I continued TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Individual Gross Pathologic Findings in the Liver - Females Group Animal Number Gross Pathologic Findings Control 106 149 160 178 200 1-3 mm dark red foci in all lobes Few 1 mm dark red foci in all lobes None Numerous 1 mm red foci in all lobes Few 1 mm red foci in all lobes T-I 303 349 369 381 397 Few 1 mm dark red foci in all lobes 1-3 mm dark red depressed foci in all lobes 1-3 mm dark red foci in all lobes 1-3 mm dark red foci in all lobes Few 1-2 mm dark red foci in all lobes T-II 524 550 569 594 599 Few 1-3 mm dark red foci in all lobes 0.5 cm dark red nodule in 1 lobe 1 mm dark red foci in all lobes 1-2 mm dark red foci in all lobes 4 cm subcapsular nodular focus in major lobe, capsular adhesions to diaphragm, spleen and serosal surface of small intestine T-III * 717 728 732 735 742 0.5 cm dark red focus in 1 lobe with capsular adhesions to mesentery None 1 cm dark red focus in 1 lobe 3 cm nodule in 1 lobe; 0.5 cm dark red nodule in another lobe; focal capsular adhesions between nodule, small intestine, stomach, spleen and mesentery 4 mm dark red depressions in 1 lobe, 1-5 mm dark red foci in all other -lobes CMA 102263 CMA102264 Organ Lung Kidneys Ovaries Adrenal glands Uterine horns TABLE II TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Summary of Gross Pathologic Findings in All Tissues Except Liver Findings Group: Number per Group: Control 10/10 Male/Female Incidence T-l 10/10 Male/Fern ale Incidence T-II 10/10 Male/Female Incidence T-III io/io b i Male/Female \ Incidence ^ Focal adhesions to diaphragm or thorax Few pin-point red foci in all lobes Focal consolidation of 1 or more lobes Abscesses in parenchyma 3/0 1/0 2/0 0/0 1/0 0/0 0/0 1/0 0/1 0/0 0/1 1/0 0/0 1/0 0/0 0/0 Unilateral hydronephrosis Bilateral hydronephrosis Unilateral or bilateral follicular cysts 0/0 I/O 0/0 0/0 I/O 0/0 0/0 0/0 220 1 Small (1 mm) blood-filled cysts in cortex of 0/1 0/0 0/0 0/0 1 gland Enlarged and dark red color 0/0 0/0 0/1 0/0 Hydrometra 200 0 1 S3 1-019 3i/idnAi'uaL B I O - T E S T CMA 102265 TABLE II continued , TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Summary of Gross Pathologic Findings in All Tissues Except Liver Organ Findings Group: Number per Group: Control 10/10 Male/Fern ale Incidence T-I 10/10 Male/Female Incidence T-II 10/10 Male/Female Incidence T-HI 10/10 Male/Female Incidence | | v ^ Pituitary gland Spleen Hemorrhagic focus Few adhesions to mesentery Enlarged (3 times normal size) 0/0 1/0 0/0 0/0 0/0 0/0 1/0 1/0 0/0 0/0 0/1 0/0 Abdominal cavity Small amount of blood in peritoneal cavity 0/0 0/0 0/1 0/0 CMA 102266 tajvj.l m i'L'S'i' MAI'lCiilAi.: Vinyl bblur/tb.- (isJbyJem.- UeJ *v.tJ) Clnonic Vuput Inhalation Study AlhUtu Hat* One Vtjr kntciim ^uuii(k.c Group ami Animal Number bile Duct Prulliera Uoo Portal to Sinusoidal rlr)ut:tliitbry Necrosis ul SuiimMry of l.ivei -Mates Cyi<J|)taSmtc llypcrtiupby LlpUl Vacuoles* of ut lleuatucytes llupalucytca llypct ti tipliy diul llypoi|tUoia al ll* palm yU-s blllUsutUal Dilatation Subeapsular Tbrouglioul Uc^lun Only Sc cl iud Control J9 St ss 69 114 T-I 20S 2Sfc 264 2n 299 l'-II 4U4 4ii 4*9 469 4119 TUI 64>9 639 663 66ti 694 1 1-2 l 1 1 1-2 1 2/M 12 12/M 2/M 1 1-2/M l (tew) 1-2 12 l t i 12 k Clew) 1 2 4/D U lube) 1 1 2/M 2 2/M 2/M 12 W l 12 1l 1-2 1 l l 1 1-2 1 12 i 1 1-2 1 12 L 1 2/M 2 2 1-2/M 2/M 1-2/M 2/M 2 J/M 2/M 2J 1 2 * All lesions are local unless oiliui wise unJicaled iSystem mt Abhrcviulnin 1 - minimal m juvu lly 2 milt! in srveitly 3 -- iiiudei'((u in stvti'Jly 4 - market! tti uutfctily t* - Present. no u-ail M ~ mulUfutdl 1> - Jillosc * - cMlreUlc lu seVetity Cysllc Spaces (Pehosls) AnKlOsal ctJfu V iJ 1" sl al i ao *c TAtll.E 111 luiUmuil TLSl MATLKlAL. Vinyl Chloride (Ethylene Uunvuil) CltriJiiic Vapor inhalation Sludy Alhnio Hats Due-Year Inlet im SaerdUe Summary ul Livei Lesions* 1* enisles Croup aiid Animal Hite Duel huiifuiilion Pur la 1 to Sinusoidal Extramedullary Hematopoiesis Necrosis ul H cpalue y les Cytoplasmic Uypertiophy Lipid Vjt-uukb ul ill IJepaiocyles llcpalocytes llypcrliophy and Hyperplasia ul M i.palne y les Sinusoidal l)il slalom SuLcapsid at* Thruu^houl Ik |>u>ii Only Sk.-k.lluu Cunlrol. 10b 149 160 1 7 200 1 1 2/M 2 i1 1 1 1 1 Ti JO 3 449 469 1BI 497 1 1-2 (Itw) 1 1 i 1 12 12 12 12 1 Til 574 650 569 594 599 i l i I 1 W 12 i 1 12 12 1i 1 2/M 2 2/M T-Itt 717 77B 717 715 747 1 1 ) 1-3 12 1 12 1 1 1-2 I 2/M 2 1/M 2 1/M 2 J/M 2/M 2 12 2 2 2 11 loud Killed Cyclic Sparer (Pehusis) An Losai PP PP PP PP PV PP CMA 102267 AW lesions ai e local unless olhet wise indicated Grading System anti Ahht evialmns \ - minima] in severity 2 - mild in ^evenly i - moderate in scverily P - Piescnt, no grade M - imilllfocal 4 - marked in scveiily U - diUuse 5 - extreme in covertly TABLE IV BI O - T EST JzvoAatyuai, 3*2. CMA102268 rl Eii'i' MATEitlAL: Vinyl Chloride (Ethylene Uerivcd.) Chronic Vapor Inhalation Study - Albino Kalb One-Year Interim Sacrifice Incidental Histopathologic Changes m Tibbucb Other Than Liver Hales Organ Trachea Lung Heart Kidney Findings Group: Number Examined: Control IU Avcj jjjt Incidence Grade T1 10 Average Incide nee Grade T-Il 10 Average incide nee Grade T 111 10 Average Incide nee Grade Chronic tracheitis 2 2.0 j 1.0 2 1.0 3 i ./ Chronic focal murine pneumonia Focal aggregates of alveolar macrophages Acute focal bronchopneumonia 3 i.3 2 1.0 5 1. i 1 2.0 2 1.5 1 1.0 5 1.3 2 1.0 Focal myocardial degeneration and librosis 2 10 2 1.0 i 1.0 bilateral local proteinaceous tubular casts Bilateral local tubular nephrosis Bilateral focal interstitial lymphoid inllltraliuns bilateral chronic focal nephritis Bilateral local mineralized mlcmconcretions in tubules Unilateral hydronephrosis bilateral hydronephrosis 3 i 2 1 1.2 1.5 1.0 2.5 2 1.0 t 1.0 1 1.5 1 2.0 2 l.B 2 l.B l 1.0 4 1.5 3 1.7 1 1.0 Urinary bladder Proteinaceous plug in lumen Prostate gland Focal interstitial lymphoid infiltrations Testes Unilateral focal testicular degeneration Unilateral focal testicular degeneration with Spermatoceles, spoj matte giamdoums and dystrophic mineralization of some aifected tubules 2 1 .U 1 i.O 3 1.0 1 1.0 l 1.0 l 3.5 4 1.0 CMA102269 TABLE IV continued 1ESI MA i EKIAh; Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Halt. Oue-Yciir Interim Sacrdice Incidental Histopathologic Changes in Tissues Other Than Live* Males Organ Pituitary gland Finding:. Group: Number Examined; Control 10 Average Incidence Grade .T 1 It) Average Incidence Grade r ii 10 Average Incidence Grade Till 10 Average Incidence Grade Focal cytoplasmic vacuolation of basophils Small chromophobe adenoma 2 1 .0 2 1 .U 1P 1 1.0 Adrenal glands -Unilateral locus of lipomatuns degeneration in cortex 2 1.0 ) i.t> 1 1 .0 Thyroid glands Bilateral diiiuse hypertrophy of lollictiJar epithelial cells i 2.0 Colon Bone Nematode parasites hi lumen (ncmutodiasis) Focal medial hypertrophy uf aitcriolcs in femoral marrow 1 i J.O i 2.0 i 1.0 1 2.0 Grading System 1.0- minimal in seven *y 2.0 - mild in severity 3.0 - moderate in severity 4.0 = marked in seventy 5.0 - extreme in severity I? - Present no grade **v si ai *o i a ,i- 8 1 0 -TEST CMA 102270 1 Organ Trachea Limn Heart Kidneys Spleen liter us TAULli IV continued TFST MATbHIAb; Vinyl Chloride (Llhylcne Derived) Chronic Vapor inhalation Study - Albino Hats One-Ycur Interim Sacrifice Incidental Histopathologic ClidJi^cb in Tissues Oth er Than Liver - Fernat es Findings Chronic tracheitis Group: Number Examined: Control 10 Average luci ilence Grade T1 10 Average incidei iCe Giade T II 10 Average Incidence Grade T 111 10 Average Incidence Grade 3 1.8 3 1.5 2 1.8 . 1.0 IS Chronic focal murine pneumonia Focal aggregates of alveolar macrophages Diffuse hemorrhage and edema (agonal) Focal suppurative bronchiolitis Few small loci ol chionic myocarditis 5 1.2 2 1.0 1 3.0 1 1.0 4 1.0 4 1.6 1 2.0 4 1.1 1 1.0 1 1.0 Unilateral focal proteinaceous tubular casts bilateral local proteinaceous lubulai casts Bilateral local tubular nephrosis Unilateral chronic focal ucphriLis Unilateral focal mineralized micruconcretioiis in tubules bilateral focal mineralized microconcrelions in tubules Unilateral large tubular cyst in Lurtex Bilaleral ditfuse brown granular pigment and protein resorption droplets within cytoplasm ol cortical tubular epithelial Celts increased hemosiderin pigment Diffuse increased extramedullary hematopoiesis 1 1 1 ' 4 2 1.0 2.0 1.8 1.0 1.0 1.5 3 1.0 2 1.5 1 1.0 3 2.0 1 1.0 4 1.6 1 1.5 1 1.0 1 3.0 1 1.0 l I .0 4 1.5 1 1* 2 1.0 i 2.0 Hydrometra Suppurative metritis with endometrial abscesses Suppurative inelrilis 1 3.0 ' 2.0 i 2.0 i o 2> 8 TAtiLE IV continued TEST MATERIAL: Vinyl Chloride (Elhylcne Derived) Chronic Vapor inhalation Study - Albino Rais Gnu-Year interna Sacrifice Organ Ovaries Pituitary gland Adrenal glands Pancreas Thyroid gland Eye Colon Incidenial Histopathologic Changes in Tis&utib Oilier Than Liver females Findings Group: Number Examined: Control 10 Average Incidence Grade TI 10 Average Incidence tirade T-ll 10 Average Incidence Grade T 111 10 Average Incidence Grade bilateral focal follicular cy^U Unilateral focal iolliculai cysts ip ip ip 2P Focal hyperplasia of chromophobe cells Unilateral focus of lipomatous degeneration in cortex Unilateral local hematucysls in cortex Bilateral focal hematucysls ni cortex Focal atrophy of acinar colls bilateral diffuse hypertrophy ot lollicular epithelial cells l 4.0 ) 1.0 1 1.0 2P 1 2.0 2 2.0 1 1.0 1 1.0 1 i.O 1 2.0 1 2.0 1 1.0 Unilateral chrome focal dacryoadenitis Unilateral local retinal degeneration Nematode parasites ui lumen (nemalodlasis) i 1.0 1 1.0 2P Grading System 1.0 = minimal in stvtnSy 2.0 = mild in itvti ily 1.0 = model-ale in severity 4.0 = marked m severity 5.0 = extreme in severity P - Present, no grade BIO-TEST I,