Document 7R0RZ6X8VG31OeaxogDkeG966

R&S 115584 DttJUut'ual B I O T E S T J.ab&uzto'ti&i., H*tc. )/ MANUSCRIPT FOR PRESENTATION AT TIIE INTERNATIONAL ACADEM OF PATHOLOGY MEETING ON MARCH 5, 1975, IN NEW ORLEANS, ENTITLED mmmm Inc. , Northbrook, Illinois; National Cancer Institute, Bethesda, Maryland; Northwestern University Medical School, Chicago, Illinois; Mount Sinai School of Medicine, New York, New York: and Northwestern Memorial Hospital, Chicago, Illinois. Ladies and Gentlemen: Vinyl chloride monomer (VCM) is a gas at ambient temperature and atmospheric pressure. It is made by several processes that involve the chlorination of ethylene or acetylene. VCM is used primarily in the manu facture of polyvinyl chloride (PVC), a resin which serves as the parent compound for a number of plastics. In May of 1970, Dr. Viola, from Italy, presented a paper at the 10th In ternational Cancer Congress in Houston, Texas, on the carcinogenic effects of vinyl chloride (VC) in rats. He reported tumors in the skin (para-aural carcinomas), lung (epidermoid and adenocarcinomas) and bone (osteochondromas) of Wistar rats that had been exposed to 30, 000 ppm of VC vapor for 4 hours a day, 5 days a week, for 10 months. This was probably the earliest report of a carcinogenic effect of VC in experimental animals. His study was originally designed in an attempt to develop an animal model for acroosteolytis, a bone disease that was reported in the early 1960's among VCM and PVC workers. or T, Mtejr. H/fAtj77TX HtuLtibual B I O - T E S T 2ai&taivUe&. 9nc. 2 R&S 115585 In view of Dr. Viola's findings, inhalation studies in rats were subsequen^^ initiated in Bologna, Italy, by Dr. Cesare Maltoni in which vapor concentrations of 50 to 10,000 ppm were investigated. Early in 1973, it was disclos d that angiosarcomas of the liver and tumors in other organa (Zymbal's gland tumor of the ear canal and Nephroblastomas of the kidney) had been seen in these animals. At that time, liver tumors had not been reported in rats at exposures of 50 ppm of VC. In view of these findings by Drs. Viola and Maltoni, a vapor in~ halation study with VCM in mice, rats and hamsters was started in September of 1973 at Industrial BIO-TEST Laboratories, at the voluntary request of the Manufacturing Chemists Association (MCA) which represents a group of 31 companies in the United States involved in the production of VCM or FVC. The study was designed, by this group, as a life-span study in each of these species to complement Dr. Maltoni's study. The exposures selected were 50, 200 aW 2, 500 ppm of VC vapor with exposures 7 hours a day, 5 days a week. Two hundred of each species and each exposure level were used, for a total of 600 animals per exposure level. Preliminary tumor data from animals on this study were reported at the end of the first 8 months when liver angiosarcomas appeared in mice that had died at the 50 ppm and higher levels. This preliminary data was presented on April 15, 1974, to representatives of the Environmental Protection Agency (EPA), Occupational Safety and Health Administration (OSHA) and the National Institute of Occupational Safety and Health (NXOSH). Htu.Labial B I O - T E S T Jlai&tatiwU, 9mc. 3 Meanwhile, the first 3 fatal cases of liver angiosarcomas among vinyl chloride workers in the United States had been reported in January of 1974. With the identification of this rare tumor in humans and experimental animals, there rapidly resulted a national interest in this problem. Dr. Louis Thomas, at the National Cancer Institute in Bethesda, Maryland, and Dr. Hans Popper, who was at that time a Fogarty Scholar at the National Institute of Health, were asked to review all the angiosarcoma cases among workers in the vinyl chloride industry. In September of 1974, I met with Drs. Thomas and Popper to review slides from both the human and mouse angiosarcoma cases. At that time. Dr. Thomas had tumor material from 14 or 15 industrial workers. Slides of human angiosarcoma cases of unknown etiology from Dr. Godfrey Kent at Northwestern Memorial Hospital in Chicago were subsequently reviewed. Slide #1. Gaseous Vinyl Chloride-Induced Hepatic Angiosarcomas in - 15 workers in Polymerization Plants - Rodent Experiments Does Similar Morphologic Evaluation in Man and Mice Enforce Causal Relation and Permit Differentiation From Angiosarcomas of Unknown Etiology? Within 8 Months of Exposure - Hepatic Angiosarcomas Seen in - 2/200 Mice at 50 ppm - 11/200 Mice at 200 ppm - 28/200 Mice at 2,500 ppm The purpose, then of this presentation is to present some of the comparative. R&S 115586 BIO-TEST Xa&otaiMiai,, Shc. 4 early developmental and morphologic features of the human and rodent angiosarcomas that have been studied thus far. In this slide, preliminary tumor data at 8 months from our mouse study revealed an exposure-related relationship in the incidence of grossly visible liver tumors that were later verified histologically as angiosarcomas. The first hepatic angiosarcoma appeared at 6 months in 1 animal at the Z, 500 ppm level that had died. The gross appearance of these tumors in man and animals was similar. The livers were frequently enlarged and contained solitary to multiple dark red circumscribed or nodular foci. The tumor foci in mice varied in size from 1 mm to 3 cm in diameter and the cut surface contained blood-filled cystic spaces associated with necrotic foci. The tumors usually extended up to and involved the capsular surface of the liver. In most of the animals with liver tumors, there was blood present in the peritoneal cavity that resulted from hemorrhages at the capsular surface of these tumors. Internal hemor rhage was one of the major causes of death in these animals. Slide #2. This is a section of human liver in which there was an angiosarcoma in another portion. The earliest lesion that Drs. Thomas and Popper have been able to detect, and have reported, is this irregular focal sinu soidal dilatation without necrosis of hepatocytes. R&S 115587 r &S 115588 ))ttduiifUal BIO-TEST IJhc. 5 Slide #3. This is just a higher magnification of the previous field to show a few enlarged sinusoidal lining cells. Slide #4. In the human cases, there is a characteristic, progressive portal tract and capsular fibrosis present throughout the liver that can be seen in this Aniline Blue-stained section in which collagen and reticulin fibers stain blue. This feature has not been observed in liver sections from our VC-exposed animals. However, the liver of rodents (rats, mice, hamsters) normally contain less connective tissue within the portal tracts and lobules when compared to the liver of man. Slide #5. This is the earliest lesion in the mouse which again is focal sinusoidal dilatation. Slide #6. At higher magnification, some of the lining cells appear to be slightly increased in size. There is also focal hypertrophy of hepatocytes in these areas. Slide #7. There is also a focal hypertrophy'of hepatocytes in areas without sinusoidal dilatation and some of these hepatocytes are binucleated. HttJMdhkU. B I O - T E S T JaAvutfoUei, 9*c. 6 R&s 115589 Slide #8. The next stage in progression of both the mouse and human lesions is the formation of small to large blood-filled cavities or areas of peliosis. This section is from a mouse. The larger lesions are usually located just beneath the capsule and, microscopically, they are frequently early or advanced lesions of angiosarcoma. Slide #9. At higher magnification of such a peliotic focus, the neoplastic process can be seen extending up to the capsule. Rupture of the capsule at these sites may result in fatal internal hemorrhage. In addition to fresh blood, these peliotic foci contain large fibrin thrombi which are not present in this field. Slide #10. A slightly higher magnification of the angiosarcoma reveals papilla projections of liver cords extending into the peliotic space which are en veloped by neoplastic lining cells. In some areas of these tumors, there is slight widening of the Space of Disse. However, this feature is not as prominent nor as frequent as seen in man. There is also hypertrophy of the entrapped hepatocytes which later undergo necrosis. Slide #11. In other areas of the tumor, there are solid sheets of neoplastic ceils that have replaced the liver cords. Slide #12. This is the counterpart of the above liver angiosarcoma in man where we again see a papillary projection of liver cells extending into a bloodfilled space that is covered by neoplastic lining cells. There is a prominent separation of the Space of Disse which contains a non-neoplastic prolifera tion of different types of cells including macrophages, lipocytes and fibro^ blasts. DudUtibuol B I O - T E S T J,alMVialtvu&i; 9ttc, 7 Slide #13. This is another area of the same tumor to show the presence of intracannicular bile thrombi in hepatocytes due to stasis of bile flow. Slide #14. This Aniline Blue-stained section shows an increase in perisinusoidal connective tissue. Eventually, with destruction of the entrapped hepato cytes, there is extensive fibrosis in the area of the angiosarcoma. Now, I would like to briefly show you a series of slides of the developmental lesion and angiosarcoma in rats. Slide #15. Again, this is the initial lesion, an irregular focal sinusoidal dilata tion without necrosis of hepatocytes. This lesion seems to occur more frequently in the subcapsular region of the liver lobes. Slide #16. At higher magnification, there is hypertrophy of a few endothelial cells and some nuclei contain mitotic figures. Slide #17. We now see hyperplasia and hypertrophy of the lining cells which have enveloped most of the liver cords. Some of these cells are clearly neoplastic. R&S 115590 9ndu&Uial B I O - T E S T J.aivuafoUe&. 9*c. 8 Slide #18. ------------------------------------------------------------------------------------------------------------------------ +1 The hep'atocytes in the immediate vicinity of the tumor have now been replaced by these sarcomatous lining cells. Slide #19. A few large pleomorphic tumor cells can be seen in the lumen of one vessel as well as in and along the margin of the tumor. Slide #Z0. In this trichrome-stained section, there is an absence of collagenous connective tissue in the stroma of the tumor. Slide #21. In this section, tumor cells can be seen within a portal tract vesse adjacent to a bile duct. Last Slide. Common Features of Human and Mouse Angiosarcomas Support Etiology and Epidemiology: 1. Multiple, Focal, Sinusoidal Dilatation 2. Blood-filled Sinusoidal Cysts (Peliosis) 3. Proliferated Hepatocytes Enveloped by Sarcoma Cells 4. Progression to Papillary Pattern 5. Vasoformative and Anaplastic Nodules are Rare Dissimilarities: in Mice and Rats 1. Less Fibrosis (in Tumor and Parenchyma) 2. Often Uniform Sarcoma Cells vs. Multipotential Cells in Widened Tissue Space of Disse in Man R&S 115591 9ndLubual BIO'TEST 9**c- In summary, there are a number of common features between the human, mouse and rat lesions from exposure to vinyl chloride. These common or similar features are; multiple, focal areas of sinusoidal dilatation. We do not believe that these are angiosarcomas at this stage. We have no way of knowing at just what step, histologically, the angiosarcomas start and then progressively develop into a clinically important tumor. The second point of similarity is that of the peliotic lesions (blood-filled sinusoidal cysts). Then, there is, at the early stage of angiosarcoma, focal to multifocal hyper trophy and proliferations of hepatocytes, enveloped by sarcoma cells. This progresses to a papillary pattern and, finally, there may oe solid anaplastic areas within the angiosarcoma. There are a couple of points of difference: in the mice and rats, we see very little or no increase in connective tissue in the neoplastic and unaffected portions of the liver when compared with man. Finally, there seems to be a somewhat more uniform population of tumor cells in the animal tumors than in the human. In man, there is a possibility of not only the endothelial lining cells but some of the other cells in the tissue space of Disse being involved in the neoplastic process. Thank you. R&S 115592 t vH ANNUAL MEETING ABSTRACTS Laboratory Inyistigation f opposed or exaggerated estrogenic stimulation. Most organs. Nonsuppurative meningoencephalitis was pres of these cellular characteristics, with the exception of ent throughout the brain, especially in the pons, cere intracytoplasmic microfilaments, are comparatively re' bellar peduncles, and thalamus. SHF appears to be an duced in severe adenomatous hyperplasia (carcinoma excellent primate model for virus-induced disseminated in situ) and well differentiated adenocarcinoma, and are intravascular coagulation and other human diseases. ahsent in neoplasms of lesser differentiation. The grad* ual failure to express estrogen-dependent cellular alter ations in these lesions seems to he related to neoplastic dedifferentiation, and to he independent of the estradiol receptor content. The subcellular effect of progesterone or synthetic progestins on hyperplastic and well differ entiated neoplastic endometria is that of epithelial re gression and includes secretory conversion, lysosomal hydrolytic activity, a decrease in the length of micro villi, and lack of ciliogenesis. These observations sup port the concept that progestogens act upon endo metrial hyperplasia and neoplasia hy a direct, nonimmunologic, cellular effect by promoting secretion and inhibiting development of estrogen-influenced cellular specialization. Ultrastructural Changes in Endometrium of Wo men Bearing Copper Intrauterine Devices Amador Gonzalez-Angulo and Ramon AznarRamos. Departamento de Investigacion Cientifica, Centro Medico Nacional, Instituto Mexicano del Seguro Social, Mexico. D. F., Mexico. Clinical studies have confirmed that the addition of metallic copper to inert polyethylene intrauterine de vices increases significantly their antifertility action. The continuous release of copper by the device has been piwtulated as one of the mechanisms whereby con traception is attained. The aim of the present study was to investigate epithelial and stromal changes possibly related to copper deposition. Endometrial biopsies were obtained from 12 young women bearing TCu-200 intra R&S 1155 The Pathogenesis of Simian Hemorrhagic Fever: Hematologic and Histopathologic Studies W. E. Giddens, Jr., and D. Jessup, H. E. Branson, L. A. Mayer, D. R. Benjamin, and W. T. London. Departments of Pathology and Lalwratory Medicine, the Regional Primate Research Center. University of Washington, Seattle. Washington. 98195, and the National Institute of Neurological Diseases and Stroke, National Institutes of Health. Bethesda. Maryland 20014. Simian hemorrhagic fever (SHF) is an acute viral dis ease of monkeys of the genus Macnca. It has caused several epizootics in primate colonies around the world, resulting in the loss of thousands of monkeys. To under stand more about the pathogenesis of this disease, we gave five rhesus monkeys intravenous injections of a 10"4 dilution of serum containing the Corbel strain of SHF virus. Blood was taken before inoculation and at uterine contraceptive devices for more than 6 months. Six biopsies were taken at day 10 and six at day 20 of the menstrual cycle. The main changes were located in the cell organelles at day 10 of the cycle. The mitochondria disclosed vacuolation of the matrix and myelin figure for mation in 70 to 80 per cent of the epithelial cells of lour patients. There were also increased numbers of lysosomes and hyperplasia of Golgi structures. Stromal capillaries showed myelin figures in endothelial cells at day 10. Only in one case of secretory endometrium were similar alterations present. Preliminary studies hy energy-dispersive x-ray analysis (Kevex) failed to reveal copper in various cell organelles. The above observa tions seem to indicate that there is a definite altera tion of mitochondria of epithelial cells which may result in impairment of respiratory mechanisms and energy production, rendering the endometrial environment in hospitable for the fertilized egg. selected intervals post inoculation (PI) for hematologic, ^Comparison of the Morphologic Features of Hepatic ultrastructural, and coagulation studies. Monkeys were Angiosarcoma in Man and Rodents following subjected to euthanasia at .'1. 5. and 7 days PI; one Prolonged Exposure to Vinyl Chloride monkey died spontaneously at 7 days PI. I). E. Gordon, L. B. Thomas. J. C. Oalandra. H. Disseminated intravascular coagulation was a prom Popper, and G. Kent. Industrial Bio-Test Labora inent feature of the disease and was characterized hema- tories, Inc.. Northbrook. Illinois. National Cancer tologically by markedly decreased plasma fibrinogen Institute. Laboratory of Pathology*. Bethesda. Mary levels and increased degradation products of fibrinogen land, Northwestern University Medical School, Chi and fibrin. Platelet counts were reduced to about one- cago, Illinois. Mount Sinai School of Medicine. New half of the preinoculation values. Clinically, there was York. New York, and Northwestern Memorial Hos hyperthemia. anorexia, weakness, epistaxis. and dys pital. Chicago. Illinois. entery. At pecropsv the blood often tailed to clot, and A' causal relationship has hecn established between there was hemorrhagic enterocolitis, lymphadenopathv. the development of hepatic angiosarcomas among work splenomegaly, and widespread pctcchiae and ecchy- ers engaged in the polymerization of vinyl chloride. The moses. On histopathologic examination, there was ne histologic features of similar hepatic tumors that de crosis of lymphocytes in the centers of the Malpighian veloped in rodents (mice and rats) within 1 year after corpuscles of (he spleen and in the germinal centers of exposures of 2500. 2fK). and 50 p.p.m, of vinyl chloride the lymph nodes and peribronchial lymphoid follicles, lor 7 hours a day, 5 days a week, were compared with in the Fever's patches and the lamina propria of small 14 cases of those that have occurred in man. To date, and large intestine, and in the thymus. A consistent the incidence of hepatic aiigiosarcoma in rodents has lesion was engorgement of splenic sinusoids with fibrin, been related to the level of exposure to vinyl chloride. and fibrin thrombi were present in many small arterioles Despite dissimilarities between rodent and human le and venules in the intestinal mucosa, lung, and other sions, such as minimal fibrosis in mice, the basic evolu- Vnl.32. No. X 197S ANNUM. MEETING ABSTRACTS 9 R&S 115594 tion was the same. The earliest lesion was focal dilation placenta with nitro-blue tetrazolium solution, an oxida of sinusoids in which the lining cells were increased in tion-reduction indicator which yields purple formazan number, piled up. and exhibited hyperchromatic and pigment at sites of dehydrogenase activity. Areas of bizarre nuclei. Subsequently, sarcomatous lining cells dehydrogenase deficiency had a white to yellow-whitj enveloped plates of hepatocytes which were hypertrophic appearance and were readily differentiated from adj^| and hyperplastic. They become cords surrounded bv cent purple-stained tissue in which formazan pigment proliferated lining cells of different types* in papillary was deposited. Succinic acid dehydrogenase activity arrangement associated with blood-filled cavities re was present in the placentas from uncomplicated preg sulting from sinusoidal ectasia. Event unllv. nodules of nancies, whereas placentas from complicated preg anaplastic sarcomatous cells with vascular spaces de nancies showed slight to marked decrease in formazan veloped with hemorrhage and necrosis. This charac pigment. In these placentas, addition of substrate did teristic evolution common to animal and man indicates not enhance the intensity of formazan pigmentation. a specific and identifiable effect of vinyl chloride or its Placentas of infants with low Apgar scores and placentas metabolites upon hepatic mesenchymal and epithelial associated with neonatal morbidity and mortality con cells. It enforces the causal relation of vinyl chloride ex sistently demonstrated reduced enzyme activity. The posure to angiosarcoma formation, but also assists in the clinical diagnosis of uteroplacental insufficiency was exclusion of vinyl chloride as the cause of some angiosar also associated with absence or marked decrease of suc comas of different appearance. cinic acid dehydrogenase activity. (Supported by Pro fessional Stall Association. The Los Angeles Countv- Ultrastructural Changes in Pulmonary Bleomycin llniversitv of Southern California Medical Center. Proj Toxicity ect 2-165-0-C.) F. Gyorkey, I. Daskai, and P. Gyokkky. Veterans Ad ministration Hospital 'and Baylor College of Medi Acquisition of Columnar (Barrett Type) Epithelium cine. Houston, Texas 770.'ll. Ultrastructural studies of the nuclei of type I and type in the Distal Esophagus after Partial Esophagogastrectomy II pulmonary epithelial cells and of alveolar septal cells Stanley R. Hamilton and John J. Yardlf.y. De from patients with diffuse pulmonary fibrosis following partment of Pathology. The Johns Hopkins University bleomycin treatment revealed that the drug had a dif School of Medicine and Hospital. Baltimore. Mary ferential effect on the fine morphology of these cells. land 21206. The most striking alterations were observed in the The origin of the columnar cells in the columnar epi type I alveolar cells; their number was markedly de thelium-lined distal esophagus (Barrett esophagus) is creased. with formation of nucleolar fibrillar centers and unknown. It has been proposed, however, that the a large increase in nuclear bodies. These nuclear bodies lumnar epithelium may he: (1) congenital, resulting fri^B ranged in size from 0.2 to 0.7 am. in diameter and were incomplete replacement of embryonic columnar epithe of three main varieties; membranous, beaded, and gran lium; or (2) acquired, being a response to chronic gas ular. Some nuclei contained up to 12 nuclear bodies. tric reflux. There was an abundance of nuclear bodies in all of the We have studied at autopsy a 74-year-old male who type I cells, which contained nucleoli with well defined had undergone partial esophagogastrectomy for squa fibrillar centers. The nuclei of type II alveolar cells mous carcinoma of the distal esophagus 6 years earlier. (granular pneumonocytes) contained compact nucleoli The body of the stomach, with fundic mucosa at the with ill defined fibrillar centers and few. if any. nuclear margin, had been anastomosed to the remaining normal bodies. When present, the nuclear bodies were of the esophagus. Postoperativelv there was persistent gastric membranous variety. The nuclei of the alveolar cells reflux. Following death from cardiopulmonary disease, were similar in appearance to those of type I cells but a mild stricture and a 0.6- to 1-cm. circumferential zone did not contain nuclear bodies of the granular variety. of tan mucosa was found in the distal esophagus just We postulate that (he ahove described ultrastructural above the suture line. There was no tumor in the area changes, in particular the appearance of fibrillar centers of resection. The circumferential zone resembled mu and nucleolus-derived nuclear bodies, were induced by cosa of a Barrett esophagus, being composed of co bleomycin. lumnar. goblet, and endocrine cells. The columnar cells of the_ surface and glands showed periodic acid-Schiff A Macroscopic Method for Evaluating Succinic Acid (PAS)-positive and Alcian Blue (AB)-negativc mucin. Dehydrogenase in the Placenta Scattered goblet cells stained with both PAS and AH. T. D. Haij, B. A. Woom.iNG, N. E. Warner, S. B. Numerous argyrophil cells and scattered argentaffin Furukawa, and H. W. Puffer. Department of Pathol cells were demonstrated. ogy. University of Southern California School of Med Findings in this case strongly indicated that the co icine. Los Angeles. California. lumnar epithelium in the distal esophagus was acquired, A histochemical assay for succinic acid dehydrogenase perhaps stimulated by gastric reflux. It has been sug described for evaluating respiratory enzyme in myocar gested that Barrett epithelium may arise from cardiac dium (Nachlas, M. M., and Schnitka. 1'. K. Am. mucosa. This case also indicated, therefore, that the Pathol. 42: 279( 1962) was adapted and modified for the presence of cardiac mucosa is not necessary for fnra^ study of 106 placentas from normal and complicated tion of columnar epithelium-lined esophagus. alth<^P pregnancies. The method entails incubation of the its origin remained unclear. Metaplasia of squamous