Document 5bOjZmazY8631B6pwbX0dZ085

fe Cc /? t i* i NKt;^T9/4IMMKn39tt| NS/0 t MWHKWHMi b*vM*m*-nni* l . t . nihl V f`MakT >k* **`~*-- <----- >>t--- r **-- *-* * *--1 y -> n-.>--t-jQ . ----------- Voi. 40. N 3. p ;t7n, IP7P .. - |f N A Polychlorinated Biphenyl-Induced Morphologic Changes in the Gastric Mucosa of the Rhesus Monkey Gregory M. Becker, M.Sc., Wilbur P. McNulty, M.D., and Mary Bell, Ph.D. Laboratory of Pathology, Oregon Regional Primate Research Center, Beaverton, Oregon 97006, and Kettering Laboratory, Department ofEnvironmental Health, University of Cincinnati, Cincinnati, Ohio 45267 The early morphologic events in polychlorinated biphenyl-induced lesions of the gastric mucosa of immature male rhesus monkeys {Macaco mulatto) were studied in serial biopsies taken during consumption of diets containing 3. 10, 30, and 100 mg. per kg. <p.p.m.) of Aroclor 1342. The severity of the lesions correlated directly with both the duration and the level of exposure. The first changes wore see* after Ingestion of 30 and 100 mg. per kg. for 3 weeks; changes were not seen until 10 . weeks at 3 and 10 mg. per kg. ' * Hie principal effect was an apparent arrest ofthe differentiation ofgenerative cells of the isthmus and neck regions into parietal and symogenic (chief) ceils. Mature parietal and zymogenic cells were found only in the bases of the glands, where the oldest cells are normally found, and these shewed sign* of injury including dilation of the rough endoplasmic reticulum in zymogenic cells, irregularity of the mitochondria in parietal cells and irregular luminal membranes, and an increase hi the number of large autophagic vesicles in both cell types. Additional key words: Gastric epithelial cell differentiation. Parietal and zymogenic ceils. Cell Injury, AuUiphagocytosla, Macaco mulatto. Polychlorinated biphenyls (PCBs) are widespread en vironmental contaminants of industrial origin (48); they enter food chains, and in experimental diets, induce a variety of pathologic changes (19). In rats, mice, and rabbit*, Uicm change** nrc primarily limited to the liver nd include arienofibrosis and hepatomas (35), megaJo* bepotocytosis (37), carcinomas (29), alterations in hepatic endoplasmic reticulum (44), and elevation of certain hepotk microsomal enzymes (11, 40). The dietary levels riffriil to produce these lesions were 100 mg. per kg. (ptp.m.) or more for many months. In contrast, rhesus monkey* (Macaco mulatto) develop different pathologic rhangrn at much lower dietary levels. The sebaceous glands of the skin underwent squamous metaplasia with keratin cyst formation (acneiform lesions) at a dietary level of 23 mg. per kg. of PCBs within 1 to 2 months (1) and bypirplmis of the gastric mucusa with extension of (be gastric glands into the submucosa with formation of aabmuroaal cysts was observed at 25 to 300 mg. per kg. af PCHs at 2 to 3 months (1,3). Although similar gastric Mann have occurred after ingestion of diesel lubricating ad (41) or ingestion of toxic fst (2), which was suboeqpatfMJy found to contain 1,2,3,7,8,9-hexachlorodibenzopdasin (13), or have accompanied nematode infestation this change has been olwerved so consistently with PCB poisoning that we believe these compounds are also causative agents of this alteration of gastric mucosa. However, previous studies (1,3) have not dealt with early morphologic changes at the cellular level induced by PCBs, nor with changes caused by PCBs at lower con centrations that may occur in food (34, 36). 'I'herefore, in order to expand our understanding of the effects of PCBs on gastric mucosa, we obtained biopsy specimens from rhesus monkeys before placing them on PCB-containing diets, and at monthly intervals thereafter. We report here the ultrastructural characteristics of the minimally iden tifiable alterations of the mucosa. MATERIALS AND METHODS Six young (7 to 6 months old) male rhesus monkeys, each weighing 1.3 to 1.9 kg., were individually caged and fed a diet cake composed of ground Purina monkey chow, vitamin supplements, banana, water, and PCBs (Aroclor 1242, Monsanto Company) diluted in com oil. The mon keys were fed diets containing either 0, 3, 10, 30, or 100 mg. per kg. of PCBs (two were fed 10 mg. per kg.). The diet cakes contained only 40 per cent water, approxi mately half as much as natural food, and therefore the corresponding levels in moist food would range from 1 to 30 mg. per kg. A young monkey eats approximately 40 gm. of diet cake per kilogram of body weight per day. Therefore, the diets represent approximate doses of 0.12, 373 374 BKCKKH. McNULTY. AND BKLL LaMOHATONV IWVKKTICAnOW 0.4,1.2, and 4.0 mg. of PCBs per kilogram of body weight frway----------------------------- --------------------------Klfi|XH-*haped biopsy specimens, 1.0 by 0.3 cm., which included the entire thickness of the stomach wall, were taken at laparotomy under Fluothane anesthesia. The specimens were taken from the body of the stomach near the greater curvature, where the histologic appearance of the gastric mucosa was previously found to be rela tively constant in control animals examined at autopsy. These specimens were obtained at the start of the exper iment, after 12 days, and at monthly intervals thereafter for 6 months, or until the animals died or were killed for autopsy. For 24 hours before each surgical procedure the animals had no access to solid food but were allowed water ad libitum. The first biopsy specimen from each animal, obtained just prior to the beginning of the PCBrontaminated diet regimen, yielded base line data on the normal structure of the gastric mucosa. One animal fed a diet without PCBs served as a control for possible effects of surgery on the gastric mucosa. 'Hie specimens of gastric mucosa were fixed either directly fear 2 hours in Pelade's buffered I per cent O1O4 (45) or overnight at 4 C. in a solution containing 2 per cent paraformaldehyde and 2.5 per cent glutaraldehyde in 0.03 m phosphate buffer (pH 7.3). Aldehyde-fixed specimens were then rinsed in 0.03 M phosphate buffer and poatfixed in 1 per cent. OsO< in 0.06 M phosphate buffer for 2 hours at 4 C. Specimens were alcohol' dehydrated through an ascending series of ethanol to propylene oxide, and infiltrated and embedded according to Spurt (50). Thin sections for electron microscopy were placed on 200-mesh copper grids, doubly stained with lead citrate and uranyl acetate, and examined with a Philips EM-200 or EM-300 electron microscope. For light microscopy, adjacent thick sections (1 pm.) were stained with 1 per cent toluidine blue and 2 per cent borax. In addition, aldehyde-fixed, nonosmicated tissue as above was embedded in glycol methacrylate (7), cut at 2 pm., dried on slides, and stained with toluidine blue or with periodic acid-Schiff stain for the demonstration of mu cus-positive sites. The number of parietal cells in 0.11-sq. mm. fields centered over longitudinally sectioned gastric glands was determined for 10 fields from each biopay specimen in order to estimate the relative numbers of these cells. All animals died or were killed when moribund and were thereafter examined by conventional autopay pro cedures. Inasmuch as this was a study of biopsy specimens from living lean young monkeys, adequate samples of adipose or liver tissue for correlative measurements of tissue PCB levels were not obtainable. Measurements were made from tissues obtained at autopsy, but some of these deaths occurred some lime after the biopsy series was completed, and thus do not correlate with the morpho logic changes observed in the biopsy specimens. poorly defined change in Ue folds of the suhorbital final Skin. This sign was soon followed by thickening and reddening of the eyelids. Although these facial changes were evident by the end of the 2nd month in animals led diets containing 10,30, and 100 mg. per kg. of PCBs, they were not apparent in the animal fed 3 mg. per kg. of PCBs until the 6th month. Soon after these facial changes were noted, the animals fad PCB diets became lees active, and their fur beeasse rough and dull; however, there was no detectable lom of hair. All animals abruptly became lethargic and semiconmtose 1 day before death. One animal fed 3 mg. per kg. of PCBs survived 245 days whereas an animal fed 100 mg. per kg. of PCBs survived 137 days. One of the anamsAa fed 10 mg. per kg. and an animal fed 30 mg. per kg. of PCBs were killed for autopsy on days 146 and 92, reaptctively, when death was imminent. Another animal fed 10 mg. per kg. of PCBs for 189 days was relumed to s normal diet on day 169; this animal is still alive and appears healthy 1 year after exposure to PCBs cessed. Although young male rhesus monkeys at this age nor- RESULTS Clinical Observations For the first, few weeks, the monkeys eagerly ste the food cakes containing PCBs and remained apparently well. The first recognizable evidence of illness vs a Khj. I. Ijghi mk'rtirraph of the fabric mucoaa from Iter tody of the Momtrh of a normal rtirawi muntt^, Swikr wwrw trih w hr MrMiTwH by ihr dark mawua> rramuiatiwi of ownw is Ihr apical itfion of each ceU. larRv ovoid imtv tdb aro pacwlal tSc; tMa with varooiated cytopiaum in tht bottom onr-third of ibr hwdir ftanSamm tymopmiir crib. Spurr'a epoxy arrl ton. !*.; Hdnidmr bier, x ISO HONS 001444 V<4.. nrm GASTme MUCOSA IN I'OLYCIILOmNATKU llU'HKNYl. POISONING 375 TMM.R I. I'amittai. Crm. I-one artrr Frr.nmr. PCB Durr to a Nnrmw Monrrt" lltw mI mi twilim* i'iniil rHh/ftrM' 0 44 77 107 143 IN 3ft 4a. of itcwtry^ 134 7 147 I 0.4 *3 3S 4 ft I ' Aatonal wan M 10 mg. per k|. of PCB. ' Uaya the eenanl had lam fad PCB dial prior to surgical Mopv, ' Tm maumtial (Wide af gaatnr mucaee, 0.11 aq. mm. in area, war* reeatad Nam rarh gastric btopity. *'TMa binpey wee ablainrd after the animal had bean fad tha PCB drt hr 100 dayn and then returned to the control diet for 3ft days prior In aniphial biopsy. decrease in hemoglobin of I to 2 gm. per HL from the initial levels of 12 to 13 gm. per ril. No serum chemistry measurements were made. Morphologic Observations Before the animals were exposed to the experimental PCB diets, biopsies of their gastric mucosa were normal at both the light (Fig. 1) and the electron (Figs. 4,5, and 111 microscopic levels. The structure and ultrastructure of normal mammalian gastric mucosa have been exten sively studied in numerous mammals including man (16, 26, 27, 30, 38, 49, 51) but not in the rhesus monkey The gastric mucosa obtained from the body of the stomach contained typical gastric glands, frequently referred to as fundic glands, which demonstrated all of the major gas tric epithelial cell types: surface mucous cells, mucous neck cells, undifferentiated cells, parietal (oxyntic) cells, zymogenic (chief) cells, and argentaffin cells. Another type, fibrillovesicular cells, described in dogs and man (21, 31), was not observed. The normal distribution ef Pm. t Light miaiagiaph of tha paatrir mnuosa from the body af tha atamaah of a rheana manhey fad a dirt containing 10 mg. par kg. of Pda far 143 days. Only on occasional parietal cott to aoan. Spurr'a agony inliw, I urn-i tahiidina blue; xlfiO. maMy gain 100 pm. per month, all of the animals fed PCB diets neither pained nor lost weight throughout the ex periment, mn at the lowest done of 3 mg. per kg. The animal fed 10 mg. per kg. and returned to a normal diet began gaining weight again within 1 month. These ani mals dsvoloptd a persistent leukocytosis of 10,000 to 90,000 leukocytes per microliter, and each showed a Pie. 3. Light micrograph 4*momtrating wKnacij changan in tha gaatric mucosa lakon at aatopay from a rhaaus manhey M a dtot containing 10 mg. par kg. of J'CBa until ita death at I4 daya. Cyatic and rblaled gaatric gland* have invaded the muacalarm mneana to farm whom filled cyata and glandular atrveturea. Clanda and cyata are Stead with tmicua-eacretmg columnar epithelial ceito. No tymagenn1 or partatal cdb ware absarract Glycol methacrylate faction, 2 ym.; tetuntoaw blue; X40. HONS 001445 379 BKCKKtt, McNULTY. AND BKJ LaMMIATOMV IWVKKrNMTMIM coll in the rhesus gastric mucosa it seen in Figure 1. The pwlrir kIkikI can be divided into the uppermost gantrir pit or foveota, the isthmus, the neck, end the base (S3). The foveoJa, which begins at the surface and extends hastily to the uppermost extent of the parietal cells, contains only surface mucous cells. The isthmus is bounded by the up|>ermost extent of the parietal cells and the lowermost extent of surface mucous calls and therefore contains only parietal and surface mucous cells. The neck portion of the gastric gland contains both mucous neck cells and parietal cells and is demarcated by the extent of the mucous neck cells. The base of the gastric gland contains parietal and zymogenic cede, al though in llte rhesus monkey the relative number of parietal cells ia small in comparison with their population m other species, notably rodents. Argentaffin cells occur throughout the gastric gland except in the most superfi cial aone of the foveota. After the PCB-conUining diets had been initiated, biopsies showed a dramatic decrease in the numbers of parietal cells (Table 1) with a concomitant increase in the numbers of mucous neck cells (Fig. 2). This change in the tissues from animals treated with PCBs was first observed on the following days after onset of nposiw day 12 (one animal fed 30 mg. per kg. and one animal led 100 mg. per kg.), days 69 and 77 (two animals fed 10 mg. per kg.), and day 71 tone animal fed 3 mg perkg ); earlier samples had shown normal mucosa. As the treatment with PCBs continued, the neck region of the gastm glands increased in extent and became nearly devoid d parietal ceils (Fig. 2). Mucus-secreting cells could be identified by the dark tohaidtne bhae-sUiimtg granules m their apical portions which were periodic arid-Schdf' positive and electron-opsque (Fig. 6) After treatment with PCBs, the lumina of thebase and neck of the gastric glands became conspicuously ddatsd (Figs. 2 and 6). Also, the mucous neck cells were mere numerous than in the control samples taken prior to PCB treatment, both because of their exclusive occupation of the mucous neck zone (which is normally occupied both by mucous neck and parietal ceils) and because of the expanded distribution of the mucous neck cells toward Pm. 4. IkklWi'lv widjUrrml wlwi mut uun writ ivlti from i mwmal Mnirrnlr! rhrww munkry. Klrrtrwi-upw|uc, pri'Mimahly whom rowlaMag. wnfa'lrft arr prowM ia the spiral rvniwa *f iNm calls. Only a irw ikflwa kxttH tubuluvcnirlaw, mmm of which i-aSiin parlirlaa |mwt h* wen in th ntinw 4 IIkm1 ttik. Ci>Hk ihM |nmh mum uf lhr*r lutwUivyMM-trft (Kit. ty or apporrMty umhngMag mtar to parietal raNa. X1S.UU). HONS 001446 Vat4M. N. X WW CAHTKIC MUCOSA IN I*OLYC!tl*OIUNATKl> HIPI1KNYI. POISONING Ar Uw of (he glands where they were wen in position* aanuafty orcopied by zymogenic cell*. These cells were aNfphalngtrally similar to mucous neck cells seen in the DMreso neck region in (he control specimen* (Fig. 4) swept that features possibly associated with differentia tion into parietal cells, such as the formation of large namhecs of tubafovesirks as seen in control specimen m* &k ware not observed. Frequently, mucous neck nib ham treated animals (Fig. 6) showed markedly tersgalsr apical surfaces and a nearly total absence of ninsnli, as well as a disorderly configuration of the entire rei body. Muctm* neck cells were seen in mitosis in bath control and PCB-diet biopsy samples (Fig. 7). These cells possessed secretory granules in the cyto plasm, bat were virtually devoid of endoplasmic reticu lum ami Golgi bodies; they possessed few lysnaomes and tabalavesicles. lywiflHic cells were both reduced m number and ahrwtructurally altered in the biopsies from PCB-fed Nimb Marked dilation of the rough endoplasmic retic ulum waa a change frequently observed as early aa day 77 at a dietary level of 10 mg. per kg. of PCB (Fig. 8). Later specimens showed very large autophagic vesicles fptgs. 9 and 10) containing degenerating zymogen grmn- 377 Pm. 1 Macaw nock eatts in the nock region of a gastric ghnd from the body of the alomach of a rhaow monkey fed a diet containwe 10 mg. per kg. of PCBa for 18S day*. Apical regions of thaaa cd are markedly irtepalar. mkrwitti are leas munertma iHan narnad. and kmwn is markedly dilated. xtJOO. Pls. ftt As^rsi rrgise at mwctMM wck crH from s auvmal mtnatid asMonmkry. Tbs cd so hf uMicfUMinabfviHWwii to s psnvlil ag lasMwrb ss S caliia MNnmm lubvIovniHni Asms ishib nMmmwMn t-lrsr wsrlm other* contain partirolaie msttsr of an man 4ni*y mmiktr Is dial of ISr cytupiiwm. Hwnwliwnw and miwnsiW am rnoted w*h a efrvwrahn; microvilli of the imracetkdar NMlnt at mature parietal tvNa da net hove a dininwMt coat Psmpam wUh Pfc. 11). ulna, mitochondria, myelin bodies, and clear vesicles of undeterminate origin. Apparent fusing of zymogen gran ules with other vesicles (Figs. 9 and 10) aa weU as the subsequent appearance of autophagic vesicles in the lumina of the gastric glands occurred. Although these changes were observed either after fixation in Palade's OsOc fixative (45) or after initial aldehyde fixation fol lowed by 0s0 fixation, they may, nevertheless, be fixa tion artifacts. No parietal cells were seen in the isthmus or mucous neck zones in the gastric glands of animals fed 90 or 100 mg. per kg. of PCBs for 12 days nor m the gastric gHnhr of animals fed 3 or 10 mg. per kg. of PCBs for 70 days; however, some parietal cells persisted in the base of the glands. These remaining parietal cells were abnormal (Fig. 12); their luminal surfaces wars irregular, their intraceUttlsr canahculi were distorted, and their mito chondria were elongated and irregularly shaped com pared with those of normal parietal cells (Fig. 11). The argentaffin cells appeared normal throughout the exper iment. These early changes were followed in all PCB-treated animate by total mucous conversion of the gastric epithe lium, with downgrowth of the gastric glands through the mtMCularie mucosae into the submucoaa, and eventually with the formation of cysts lined by mucus-secreting columnar epithelium cells (Fig. 3). The advanced lesions HONS 001447 HONS 001448 ftj, m, N X 1V79 GASTRIC MUCOSA IN POLYCHLOttlNATKO BIWIKNYL POISONING 37* flK, ft ZyemsMik nib after 19* diy at a dietary level of 10 mg. of which may be the remain* of an ontiro eell. Somo of th* lyaawn pwlftrf fCIa Utft aahtphatic vtaiulM are clearly mn, tha hr|W granuha a*paar to ba faaia* with tha autophagic "aaklaa. x 11,000. wtrt Identical with those reported by Alien end Norbeck Thee* lesions were confined to the stomach, inaa- nidi as no chenfe* were observed elsewhere m the glrsintilinnl tract of those animals that were autopied. DISCUSSION Our serial biopsy specimens from animals fed diets containing PCBs have conformed and extended earlier reports by ASen and Norback (1, 3) of hypertrophic pattie nsocoaa compoasd of elongated hyperplastic jtedrcontsMif mucus-secreting cells in rhesus monVymwri PCBs. Moreover, the present data indicate that ITf! amoristsrl histopathologic responses can occur Mittchronic exposure to very low concentrations of these compound*. (The dosages used by Alien and Norback in (hair previous studies (1, 3) were relatively higher--25, MA.ewd300 mg. per kg.) These changes include a possible Rtf germinative cells to differentiate into parietal and zymogenic cells; furthermore, they include ultrastructural changes strongly suggestive of cell injury and eventual cell death in the remaining population of rec ognisable zymogenic and parietal cells. Replenishment of gastric epithelial cells occurs primariljrby division of relatively undifferentiated cells in the isthmus and mucous neck zones of the gastric glands (14, 24, 25, 28, 32, 33, 39, 43, 46. 49. 52-54), although immature parietal cells have been shown to have seme limited proliferative capacity themselves (14) and zymo genic crib may be responsible for their own renewal (14, 25, 29, 54, 55). The newly formed cede subsequently migrate in a "pipeline" manner (24, 25,32) toward either the lumen of the stomach or toward the bases of the glands. Vacancies are created at the surface of the mu cosa by extrusion (23) and by as yet undeecribed mech anisms at the bases of the glands. Hatton and Fujita (25) suggest that the downward migration of parietal and zymogenic cells from the generative region is more com plicated than a simple pipeline system because random MOMS 001449 3M BtiCKKK, McNULTY. ANll BBLL IjtHUNATONV iMVKMTHiATtnM cel) loan also occurs throughout the length of the bane region of the gastric glands; they propose the term "am* chasiic flow system1' to describe this mechanism. Nev ertheless, ceils furthest from the generative aoue ate generally the oldest cells and those closest to the gonerstive zone are the youngest ones. According to some investigators, rodent parietal cells have a half-life of approximately 23 days (46), whereas longer survival times have been suggested by others (2b). Thus, the gradual disappearance of parietal and zy mogenic cells in rhesus monkeys fed diets containing PCBs may be due to the normal attrition of these cdh and their failure to be replaced by differentiation of immature cells. However, since at the higher PCB kwh of 30 and 100 mg. per kg. parietal cells were absent at 12 days, existing parietal cells may have been converted In mucus*secreting cells. A possible failure ofdifferentiation would be consistent with our observation of only a few tubulovesicles in immature celts from PCB diet biopsies; whereas numerous tubulovesicles were regularly ofe served in immature cells in normal rhesus (Figs. 4 and $1 and in human (49) biopsies. TubulovestdeH are special ised structures within parietal ceils which are character1 isad by their proximity to canalicular or axed surfaces; their lack of ribosomes, and their frequent coutsat af Fla 10. Autophagic vouch ia axymosen eiwintwlaayipe 9. This vouch contrim former lyamgta graauha, awia li|aa ad feme bodm. x 11,000. Fm:. 11 Normal mature parietel rrH from a rheaua monkey prior to chomtria with those wen m Figure 12 TuMoveftkh*. many f n* treatment. Compare the relatively uniform appvar--et vf the mho- contain vesicle*, are wen throughout the cytoptwm xlMMK HONS 001*50 JM., N 3, IV7 GAKTRIC MUCOHA IN TOLYCHLOIUNATK!) HN'IIKNYI, IMDISON1NG Ml fm. IS. PiriMai ctfl iflar 1M day* it dietary level of 10 mg. per kg. of PCBs. Mitochondria are markedly irregular compare with Figure 11. ywtklw and mailer vesicles (Fi*. 11). Their presence in SB Immturv cell can be considered evidence that the cell Is dMfenntisUnn into parietal cell, inasmuch as they ape rawly found in other mature cell types of the gastric epithelium. However, the present study cannot clarify whsthsr the normal mitotic rate or the subsequent rate ef differentiation is altered by exposure to PCBe. Ragardbm of whether or how PCBe change patterns a# dMfc--miction, exposure to these compounds results inaltered structure ofthe remaining recognisable parietal and rymegrnir cede. Aging alone cannot explain the --rlsdty dilated rough endoplasmic reticulum and enor mous numbers of autophagic vacuoles seen in the re maining symogen-recretmg ceils in PCB-fed animals bedonum,sundariy altered cetta were not routinely ancoun- tsrsd m control biopey samples, even at the bases of the fondk gfonds where the oldest populations of these cells asenormoffy found. Altered mitochondria and canaltculi, observed in pnristsl ceUs after PCB treatment, wore not soon in either immstare or mature parietal cells in control sparimsm Because autophagocytosis is a common re action pattern of oublethally damaged crib (4, 5, 18, 20), jfo assurnnee in mature zymogenic crib in biopsies from PCB fod animals suggests that PCBs or their metabolites mof obo be injurious to mature gastric epithelial crib. Ahhough lorn of parietal and zymogen-secreting ceils has been observed m ether gastric lesions, such as atrophic pafeitb and pernicious anemia (15, 17), the pattern of ghanges in gastric mumaa that was observed in PCB-led shesua monkeys b clearly unique. In particular, the in- tcstinalization observed in atrophic gastritis (replace ment of normal gastric cell types by mucus-secreting ceUs similar to thorn seen in the intestine) is quite different from the increase in number and extant of distribution of mucous neck ceUs observed in rhesus monkeys fed PCBs. However, it b noteworthy that the advanced PCB-in* duced gastric lesions hear considerable resemblance to the giant hypertrophic gastritis seen in Menetrier's dis ease (8, 10, 12, 47) in which dilated mucus-secreting gastric glands are abo seen to penetrate the immculari* mucosa and to form cysts. The lesions of Menetrier's disease differ by their characteristic uniform involvement of nearly the entire stomach, a more regular pattern of hypertrophy of the rugae, and at bast s partial persist ence of well differentiated parietal and zymogenic cells. A certain specificity of PCB action seems evident in that only the body of the stomach, in a distinctive but terfly-shaped lesion along the greater curvature of the stomach, and not the cardiac or pyloric portions nor the rest of the gastrointestinal tract, was affected. Further more, PCB-induced gastric lesions have only been re ported in one other animal, swine (22). Previous reports of these batons have been largely phenomenologb stud ies of the advanced end stage lesions. The present study of the early morphologic changes induced by low dietary bveb of PCBe is only a first step in the study of the pathogenesis of these lesions. Further studies on the pomibb effects of PCBs on the proliferation, differentia tion, and survival times of gastric epithelial ceUs are in program. HONS 001*51 310 BKCKKR, McNULTY. and bkll Lammmtohv Imvkhtwmi Avhmnvletiftmvntir. The authors express their appre ciation to Dr. Wolf H. Fuhrenbach for his helpful advice. Dote of acceptance. November 1. 1978. Preltminery rmh were presented at the National Conference on Polychlorinated Biphenyls, Novendier 1975. Chicago. Illinois (fi, 42). Thin study is publication no. 9W from the Oregon Hrginnal Primate Research Canter. This work wan *p|urted by National Institutes of Health Grants HRUDUOand KIWfitHN. AtWrew rrftrint rihinmM to; Mr Gregory Backer. Laboratory of Pathnkigy, Oregon Regional Primate Heaearch Canter, 506 N.W. 185th Avanur, llraverton. Oregon 97006. REFERENCES 1. Allan JII; Response of the onnhuman primate to polychlorinated biphenyl exposure. Fed 1`roc ;U:1675, 1975 2. Allen Jit, Carsten* LA: Light and electron microscopic observation* in Miu nrn mulnlta rmmkeya fed toxic fat. Am J Vet Hee 28:1513, 1967 3. AHen Jit, Norbark OH: Polychlorinated biphenyl, and triphenyl- induced gastric mucosal hviwrplesia in primates. Science 179:498. 1973 4. Amide All. Jnuregui HO. Chang J. Tramp BP: Studies on cellular xuUiphagncytowi*: relationship between heterophagy and auto- phngy in HeL* cell*. 1-ali Invest 24:162,1971 A. Arstila AU, Trump BF: Studies on cellular autopbagocytoaia: the formutiun ofautophagic vacuoles in the liver after glucagon admin- miration. Am J Pathol 53:667, 1968 6. Hrtt M: Ultrastructural features-of gantric mucosa and sebaceous gland* after ingestion of Aroctor 1242 by rhesus monkey. fit Na tional Conference on Polychlorinated Biphenvl*. edited by Buckley Jl,, pgift. Washington L)C, Knvironmental Protection Agency. 1976 7. Hennrtt IIS. Wvrick Al). Lee SW. McNeil JH: Science and art in preparing tissues rmlietbled in plaalk for light microscopy, with special reference to glycol methacrylate, glass knives and simple stains. Stain Techno! 51:71,1976 8. Ilerenson MM, Banned* J. Preston JW: Menetrier's disease: aerial morphnlogiraJ, secretory and serological observations. Gastroenter- olngy 70:257.1976 9. Bonne C. Sendground JH: On the production of gastric tumors, hordering on malignancy, in Javanese monkeys through the agency of Norhtm norbli, a parasitic nematode. Am J Cancer 37:173. 1939 10. Hcilbh OB: Giant hypertrophy of the gastric mucous (Menetrier'a disease): a survey of the literature and report of a cast. J Am Osteopath Assoc 76:198, 1976 11. Bruckner, Khsnna KL, Cornish HH: Polychlorinated biphenyl- induced alteration ofbmlogtc parameters in the rat. Toxicol Appl Hiarmacnl 2*1*9,1974 ___ _ 12. But* WC: Giant hypertrophir gastritis: a report of fourteen cases. Gastroenterology 39:183, 1960 13. Cantrell JS, Webb NC. Mabia AJ: The identification and crystal structure of a hydrupericaidhwn-producing factor 1, 2, 3, 7, 8, 9- hexaehfcwodibrnso'/Mboxin. Acta CryauUogr Sect R Struct Cry*- (sliogr Cryst Chem H2&I50. I960 14. (`hen KY, Withers HU: Proliferative capability of parietal and tymogen eeHa. J Anal 120:421, 1975 15. Chiao N-K, Wekdrrrg If: Utlraatracture of the gastric mucosa in patients with atrophic gnslritia and pemiciou* anemia. Gaatroen- terubigy 59:36, 1970 16. CoqirtMi UK: 'live ultrastructure of the gastric mucosa in normal and hvimphyaeritimaed rata. Am J Anat 1)8:53,1966 17. ISNott IIL Garden H: G**trir brnpanm: an uftrastructural study with special reference to pernicious anemia. Arch Pathol 77:2U, 1964 19. fedurko MK, Hirsrh JG, Cohn ZA: Autophagic varuolen produced in vitro. 1. Studieson cultured macrophages exposed to chloroguine. J Cell Bhd :m:377. IWW 19. Ftslibero L Tosirily of chlorinated biphenyls. Annu Rev Pharmacol 14:1.19. 1974 20. Frank AL Christensen AK: Londualfon of arid phosphatase In lipofuarin granules atul poasihle autophagic vacuole* in interatitinl ceil* of the guinea pig testis. 4 Cell Biol J6:t, 1966 21. Hammond JB, Lalicur L Kibrillove*kular cells in (he fundkgkmda of the canine stomach: evidence for a new c*H Ivpe. Anat Hee 161: 393. 1966 22. Hansen LG, Wilson IJW. Byeriy CS: Effects on growing swine and sheep of two polychlorinated biphenvla. Am J Vet lie* 37:1681, 1976 23. Harding KK, Morris GP: Cell haw from normal and stressedga*tfir mucosae of the rat: an oltraatructural analysis. GasUuenaeftdagy 72.957. 1977 24. Hattori 1': On cell proliferation and differentiation of the fwdh mucosa of the golden hamster fractographic study combined write microscopy and 'If-thymirfm* autoradiography. Ceff Tisane Nbn 148:213. 1974 25. Hattori T. Fujita S: Tritiated thymidine autoradiugraphic study an cellular migration in the gastric gland of the goldew hammer. Ce8 Tissue lies 172:171. 1976 26. Helander HF: Infrastructure of fundus glands *f the amuse gastair mucosa. J Ullrastroct lies 4 (Suppipl, 1962 27. Helander H, Kkholm l{: Ultraatructure of epithelial rtte in the fundus glands of the mouse gastric mucosa. J (Sknutfruct Kes 37< 1959 28. Hunt TE, Hunt KA: Radioautographic study of the pruhfoistamiw (he stomach of the rat using tlivmtdine-11' and compound 49/68 Anat lire I42:5U5.1962 29. Ito N. Nagasaki H, Aral M. Makiura 5. Sagthara & lliran K: Histopathologic studies on liver tumorigenesw induced in anre bp technical polychlorinated biphenyl* and its promoting effect aw liver tumors induced by bcniene hexachloride. J Ned Canta* hri 51:1637. 1973 30. Ito S. Winchester KJ: The fine structure of the gmirir naucuan in the bat. J Cell Bid 16:341, 1963 .11. Johnson FU, Young BA: Undtfferentinted cefla in gastric mana J Anat 102:541, 1968 32. Ksku II: Dynamic analysis of the stomarh epithelium *H-thyme dine autoradiographic study. Arch Histol Jpn 27 223. HM6 33. Katnoka K: Electron microscopic observations on red iwlihntue and differentiation in the gastric mucosa of the mouse. Arch Wand Jpn 32:251. 1976 34. Kimbrough HD* The toxicity of pdychiorinated polycychc cone- pounds and related chemicala. CHC Crit Rev Toxicol 2:445k 1986 35. Kimbrough HD. Under HE; Induction of afcuiHinwii and Itegm tomas of the liver in BALB/eJ mice by polychlorinated Mphenyks (Arocktr 1254). J Natl Cancer Inat 53:547. 1974 36. Kofbye AC: Food exposures to pdychlonnnted biphenyls. Envriom Health Perspect 1:85, 1972 37. Roller LD. Zink] JG: Pathology of polycMerinatad bipteenyks ns rabbits. Am J Pathd 70:363. 1973 38. Kurosumi K, .Shibasahi S. Ucbida G, Tanaka Y: Electron nine scope studies on the gastric mucosa of normal rata. Ante HiatalJpi 15:587. )958 39. Lipkin M. Sherlock P. Bell B: Cell proliferation kinetic* he trip gaktruintesfiniTTfsct of man. If. Cdknnrwd in stamneh, deny colon and rectum. Gastroentertdugy 45:721.1963 40. Lillemt CL, Farber TM. Buber AM, Van Leon RJ: ffffbct off polychlorinated biphenyls on hepatic nrieruaomai eBayme* m rim rat. Toxicol Appl Pharmacd 23:112.1972 41. Luahbaugh CC: Experimental hyperplastic gastritis mid gmteir polyposis in monkeys. J Natl Cancer Inst 7:313,1947 42. McNulty WP: 1*CB toxicity in rhesus rnunhey*. hi Hrtitiu) Cue ference on Pdychlormated Biidimyis, editedby BurkJry JL pM Washinxlon l)C, Environmental Protection Agency. 1976 43. Messier B. Leblond CP: Cell proliferation and migr*lion mrevested by radioautography after injection of thyuudme-H* min nude man arid mice. Am J Anat 106:247, i960 44. Norhack 1)H, Allen JH: Chlorinated ammatk hydiucarbun mduead modification* of the hrpatk rndoplssnwe reticulum- roneenMg membrane arrays. Environ Health Persprct 1:137,1978 46. Palsrfe GK: A stody of fixation for ebretron amrixuupy, J Exp Med 95:265,1952 46. Hagins H. Wincxe F. 1tei SM. Dittbremwr M. The eeigin and survival of gast nc |>arietal cells in tbv mourn. Anut Hk M2:98 MMff 47. Reese l)F. Hmlgson JH. Doekerty MB: Gient hypirtraphe *f de gaMnc mvcos* tMeneirier's a correhrium4the numegH*- ugrs|dite. p*tlu>logii- and clmictd finding*. Am J Hnentgrnul Rad ium Ther Nml Med *6:619, 1962 48. Htsebroogh MW, <le Lwppe B: Aceumulikm of putycblommaad HONS 001452 Va 411. No. X 197* GASTHIC MUCOSA IN FOLYCIILDI'INATKI) HH'IIKNYI, POISONING 383 hinhrnvia tn ect*wy*trmw Knvirnn llrntlh IVnipeti 1:38. 1872 49. NhNnW, HornIJMXleiwrntter Mil. Jeffrie*Oil: The n>rnuil hurnnn jpatrir epithet*: a fine ntrurtural atinly. Ijd> IiwN 195HH. I8fi8 M. Hpurv AM: A i#iw-vi*runity rp*y rrwn rmlKthimp medium for Horlrun nnrnnwnpy. 4 UMrwMruct Hr* 201. I9tt# M. Stephen* It). Pfeiffer CJ: UI'raKtriK-lore of the Rawtric mucoaa of nanoal lalmratorv ferret*. J Wltnwtruet Hr* 22:4ft, I!H M. Hi iiiw CK, CP: llenewal tf Ihr muvouK irefla in the gastric amrum of the rat. Anal lire 115:231.1853 53. Watte! W. Geuz* XI. dr Kooij |X>: UllnwlnKiural and carhohy<lrl hinioehrmirnl Mndirs on Ihc tlifl'mnlialHM and renewal of mwouN ret!* in the rat giottric funittw Cell Town* Hew 178:445. 1977 54. Willem* G. Gwtanil I'. Vanwtrenkwie Y. ftettmm I': OH pupnlaiwwi kintlica of cvmogcn and parietal cell* in the MihmcH of mire. 7. Zeflforwh Mifcrmfc Anat f.'M:.VI6, 1972 56. Willem* G. Lehy T: Itadiimulngriiphtr and <piantitative Mudiea on parietal and peptic cell kinetic* in the mouse: *elet-five effect of gaatrin on parietal cel! proliferation. Gawtroenterulony 18414, 1975 ooi*53 RISC