Document Evb6zDvvzv8rON1oKY3Lj6Z3g

620 Light and Electron Microscope Study of Chlorobiphenyl Poisoning In Mouaa and Monkey Liver Manahiro Nithmtmi, M f), Fukuoka. Japan The effects on mouse and monkey liver of long-term Orel edminielretion of chloroWphenyte (CBP), 1.S mg/day or moro hove boon investi gated at selected Intervale by llgtM end electron . mlereseopy. Hepatoeyiee ol mlee eonlalned large amounts ol scidophllie materials In the cytoplasm, end felly vacuoles worn observed Inter. Fine structural changes In the hepotoeyloo eenaleted of a marked Inereooe of smooth endoplasmic relleglum, a reduction of rough ondoplaemlc reticulum, "myelin figure" formation In the cyto* plasm, end Incraese of mlcrobodfes end lysosome*. A marked Inereese in lipid dreptots was observed later. Results el viseiron mlereseopy In monkey Ihrar shewed an tooroaeo of smooth ondoplesmle reticulum In the hspetoeytoe and swelltog of Kupffcr's edit, with an Inereased number of lysosomee and vscuoto*. Judging from the findings by electron mloreseepy, cMrsctorletto lesions at liver cells were produced by sdmtols tratten of CBP. INGESTION of Hoe bran oil contaminated with chlorobiphenyl* compound fKanechlor 400) produced poisoning among tho general population in western Japan from summer to fall of 1968,1 with a total of 000 patients to date. This event sseme to be unique in the field of food poieoning. It wee found that contamination of the oil occurred by leaking of chlorobiphenyls com pound into the riot bran oil through pinholee in a pipe ueed for heat exchange in the manufacturing process. Now, one year after the incident, many patiants ate anxious about the possible presence of latent lesions in internal organa, ae well aa persistent der matological signs. Various reports of poisoning by chlorobi phenyls (CBP)*- since the first outbreak of acne-like lesions, due to high-boiling chlori nated compound in industry, was noted by Schwarts in 1938.* But all thaaa cases were due to exposure to the fumes or dust of this oompound. Several animal experiments have been car ried out to examine the toxic effects of CBP HubmlUad for publication Jan 27. 1070; accepted March IS. . From the Daparlmant of Public Health, Faculty ot Medicine, Kyuahu Univaraity, Fukuoka. Japan. Raad in part bafora tho 42nd annual maatiaf of tha Japan Sociaty of Industrial Medicine, Fukuoka, Japan, March 20. 1068. Reprint requeeto to Department of Public Health, Faculty of Medfeto*. Kyuahu University. Katakaao 1270, Fukuoka. Japaa (Dr. NtahimimD. by the oral route*1 and have revealed lesions of akin and liver try light microscopy. How ever, the limited resolving power of tight microscopy left unsolved problems of inter pretation. Moreover, these earlier reports were concerned primarily with tha effects of high levels of exposure for a short period. This paper extends these observations to the levels of resolution provided by electron mi croscopy and more thoroughly explores the effects of relatively low levels of oral expo sure for a long period. Materials and Methods Experiment With Mica---Tha ddN strain mice uaed wdra originally obtained from tha National Institute of Genetics, Mishima. Japan, and raised in the mouse colony of this universi ty for 12 years. Sixty ddN female mice ten weeke old were separated into three groups: (1) mice receiving olive oH or nontoxic rice bran oil aa a control group. (2) mica receiving 0.6% v/v CBP in olive oil, and (3) mica receiving toxic rice bran oil containing CBP at a concentration of 1,600 ppm. Tha CBP uaad was a mixture of chlorinated biphenyls contain ing 46% chlorine (equivalent to three to four atoms of chlorine par molecule), with a trove (0.01%) of naphthalene*. Each mouse was giv en 0.2 ml of its respective oil by stomach tubs every day. All animals were kept in metal cagee, in groups of two to three per cage, and wore given the commercial standard diet and water ad libitum. They were observed for toxic signs and weighed twice a week. Arah Environ Htalth--Vol 21, Nov 1970 \ HONS O0JO59 CHLOROB1PHENYL POtSONtNG--NISHiZVMl 621 Rq 1. -Ilmtocytu of control mouse that receKmd eWv* oH tor 26 wteks, Small voeuoiot wo ooon Ht cytepleem; 10% formaWohydo solution fixation (MmetexyNnwoein, slightly reduce* tram x 860). Fig 2.--Hepetocytee from control mouse that re ceived nontoxic net bron oil for 26 weeks. No re* merkaMo ebnormoUtles ore present. Densely stained messes in eeile represent aseocleted mitochondria end Eft; 4% plwtereMehyde fixation (totuMlne blue, ftllQhtly reduced from x 800). Seven or eight mico in ooch group were killed after four week*, and two mica in each group at a time woro killed at IS, 17, 22, and 26 wooka. All of thorn woro investigated microaroptoaily for tho prooonco of toxic alterations. Thoy woro killed by a aharp blow on tho hood, followed by decapitation, between 9 am and 10 am in nonfaatod condition. Subooquont to ex* anguinatfon, tho liven were immediately ex* deed and weigbod. Sample* from a constant it* on tho right lateral lob* wore procooeed for light and alaetron microocopy. Tissues for con ventional light microaoopy won fixed in neutral 10% formaldehyde solution, and paraffin* embaddad tinue wao etainad with hematoxy* 1ln*oooin. TIssum for oloctron microocopy woro immediately plaoad in a drop of 4% gluteialde* hydo In collidine buffer (pH 7.4) and cut into cubes of approximately 1 mm with a rasor Made. Small Mocks of tissue woro fixod for two hours at room temperature in buffered 4% glut* araldshyite and pootflxod for one hour at 0 C in collidine*buffered 2% osmium totroxido. Some of thoeo disuse wore also cut and ftxad in 2% osmium totroxido Hi -collidine* or phosphate buffer* at pH 7.4 for IVt to 2 hours. The fixed and washed tissue slices were then dehydrated in a graded series of ethanol and in propylene oxide prior to embedding in epoxy resin (Epon).1* Thin and thick sections of the epoxy*embodded tissues wore cut with glaae knives on an ultramicrotome. Thick <0.6p to 1.6s) Motion* were prepared and stained with triuidttne blue11 tor orientation. Thin roc* Sena of approximately 60 me wen stainad with uranyl acetate1* and load nitrate or Millonig's lead1* alone. Specimens were examined with an electron microscope operating at SO kv. Experiment With Monkey*--Five cynomol* gut monkey*, weighing about 1 kg (2.2 lb) each, and throe squirrel monkeys, weighing about 600 gm (11.0 lb) each, were used for this experiment The CBP was administered with frmh natural diet (potatoes, cabbages, carrots, bananas, oranges, apples, and a little quantity of of sasli, dried fiifcaa). The total amount of GBP administered to a cynonwlgus monkey ranged from 641 mg in 40 days (16 mg/day) to 346 mg in 239 days (1.4 mg/day). That admin* iateied to a squirrel monkey ranged from 320 mg In 46 days (7 mg/day) to 67 mg in 46 days (1.4 mg/day). They ware observed for general akin alterations and behavior and weighed weekly. Histological and electron microscopical examination were carried out in the same way as in mice. Results General Observation*.--Mice.--The throe groups of mice were norma) in appearance and behavior for the first three months ex cept for slight weight loss in two experimen tal groups. After three or four months, the group of mk receiving CBP in olive oil and that receiving toxic rice bran oil revealed slight decrease in activity, and gross changes of the skin began to appear. The macroscopi* cal changes seen of the skin during the Arch Environ Hialth--Vol 3t, Nov 1970 \ 0*3060 hums 622 CHLOROBIPHENYL POISONING--NISHIZVMI Fig 4. *Uvw wctlyo tram mouss rassweg C9P In otlvs oil for 19 onhii HyoHno granulis Mi mmN vocmoIoo art aoao In cyteplaawi. inURraMon a# mmN round oatta la nolad at upoar MC 10% twmMdMrydv aeludon dxaOon (hawialoiryllw walH, aOoMty raduaod from x 470). bran N for 13 waaba. 9ama ooMa oontain amaM aom> omo in eytoplaam and ravaal granular rtructurat 10% termaWahyda ioMlon ftxatten (hamamaypn-raaba adpblly raduoad from x 470). experimBttal period were m follows: (1) no--iKtonn ohengeu ol the akin around the eyelids, (2) erosion, ulceration, and parforaftien of eerlaps, and (9) loaa of hair, erosion, and uioaration at tha skin around tha nock, forelega, and aides of the chart. At this and of tha experiment, tha changes similar to those in experimental mioa wars saan in some control mica. In tha fanner group, however, tha changes wave much more marked. Pathological awstfnatfan at four waake showed a marked dilhranos in tha aha of tha livers between the grot** of mioa recahrfng CBP in oil and tha group receiving no CBP in oil (P<0.01). He mioa receiving CBP in olive oil and those receiving hade riot bran oil had a mean absolute Uvsr weight of 2.9 0.4 gm and Z1 0.3 gm, respectively, while the control mioa had a value of 1.7 0.2 gm. Massi raiativa liver weights ware 1L5 1.0 gm, 8.8 OJ gm, and 8.8 0.9 gm/100 gm of body weight, respec tively. No groas tissue hniunnllH-- ves noted othe then maikad liver snhugrossnt. Jfonheys. All monkeys fad CBP gradual- Arc* Srwiron J'faaUWKot 21, Nov 1970 HUNS 083061 CHLOFOBtPHENYL POISONING--NtSHIZVMI A____ --. ng 7.--Uvtf Metton 7<0 HepetocytM of mowo gtvow taste rte* bran bran oil for 26 wooM. ItepaiocyiM at* KupffW* oollo on for 26 mota. WuMrotiitt pwiwi of NgM son Fig 2.--Thwo fiom Uvor of cynoiwolgMO monitor ly lot weight. On of tho equirral monkaye who received a total of 20 mf of CBP in aix daya nvaalad palpafenl adana two daya befora death. Inrreaaeri diechargo tram the eye with blepharitia and. loaa of appetite wan noted in a oynomolgue monkey who noaivad 790 mf of CBP in aavatl weeka and wan obeerved for thiaa waaka after CBP waa topped Although two cyncmoigua tnonkeya who noaivad 122 mg of CBP in ais waaka nvaalad ion of appetite and weight loaa during the ebc waaka, they gained weight after CBP waa atoppad. Pathological oandnation juat baton or after death damonatrated enlargement of the liver, bat it waa found that pneumonia or diarrhea waa the metn oauae of daath far all monkayo. Light Mfcnaoovto Findings Ifice.--In the control mice receiving nontoakirtoa bean oil, liven ahowad nearly normal hletotoglcal appaaianoa throughout the erpartomit oept far alight appaaianoa of well cyto- Arth Environ Hoalth--Vol 21. Nov 1970 mons 0830*^ CHLOnOBIPHKNYL POISONING--NISHIZUMl iv.v - -*- -i n' , ^.y^- -y,y ^yyy^Wfo:, ' .'/>.. \ .. . . . .!'. %. : j-JF-T "'> J -' - %i* Ad 12. Pit of an tapUBtyo of aoiM 11--i at 17 aula of experimental pcriad, ahowlnf. precewoe of WO and abundant mitochondria (M); oandum taadaw (lead, reduced IWn X 90.400). . plaamic vacuolaa in H <4 the centrilobular ragiom altar 22 woaka or mora (Fig 1 and 2). Tin twpotocytto at mk moalvinf CBP In olivo oil hr bur wools inciooul in ho am contained email acidophilic granular matchala in the cytoplaam without changaa of tab* lor atructure (Fig 3). Hiia auggwtad that th* liw wlananant wao dna to nytnrltotoin " '' ` ArcA Environ HaM--Vot 11, Nov 1170 MONS 083063 Fig 13. Portion of on hoy Kyto from mouoo gtoon CIT Mi on ifwrooto* omoimt of m flog <N>. i loo (I _ ___ rodxood from x 34,300). eyto from Fig 13. Thom to "myoNo Ogoro'* (MO M wfdca mudfiooloMUr Ndy aifvtaeasiasa UMMg 0(u&UUH-U|MIUO from x 301400). Arvh Snviron H*4th--Vol 11, Nov 1970 MONS 063064 6* CHLOROBIPHENYL POISONING--NISHIZVMI asm frequently. At 13 weeks after com* mancement of this experiment, more eoalnstained hyaline grenute and small vacuoles In the cytoplasm began to be nbeenwrl (Fig 4). After 17 weeks, the nuclei of the hepetocytee revealed variation in tiae, and outlines were not dear in some cells. Stainability at cytopiaam generally deaeaesd In sections stained with hematoxylin-eoain, and the cytoplaera wae granular. Althoughlobular arddteetme was well praeerved and no necro sis wee seen, there wae remarkable variation in eiae of hepetooytea and their nudd at 26 weeks. Some Kupffer's celle slightly incrcnmrd in six (Fig 5). The Uvne of mic* receiving toxic rice bran oil showed the typical lobular structure, with hepatocytee arranged in oords radiating from the central veto. However, hepatocytee to oentrilobular areee contained anal) vacuoles in the cytoplasm at 13 weeks (Fig 6). At 22 weeks, there wee a marked increase in the number of email vacuoles, and meat of the nuclei increased In else ununiformly. After 26 weeks, sinusoidal endothelial cede, Kupffer's cells, and some hepstocytee around the cen tral vein contained brown pigment in thenenlarged cytoplasm. Hepatocytee and Kuptier's cells increased to dee and had a reticu lated architecture in their cytnpleam (Fig 7 and 8). Monkeys.--Histological findings in livers of monkeys fed CBP generally revealed the presence of enlarged Kupffer's cells, with vacuoles to the cytoplasm of hepatocytee. In a squirrel monkey that reosived e total of 61 mg of CBP pm kilogram and died eight days from oommsuosmant of the sapertment, Arch Envinm *-Voi 21, Nov 1979 Q83Q65 HONS CHLOROB!PHENYL POISONING--NISHiZUMI 627 -,..s viy-** >* ~p 'V- :;N. v 1 k*-- .J..yr. . ' Z\ J Vi! L'f Ifcjv ' *k ' ' *'V* - /-.x .1 r^v;;. r*j> ' -I. ; ' > tmJ* A ' ' 4*-f. i .V`: V/':X^ - .*:. *.v r *.> :; `-J` **: V'*vv*A' '*A''. >r.u_- \V.. V. .< i- V- ;/VV-y;, . ,*i :j Fig 18.--Pert of n hepetoeyte from mouM given toxic rice bren oil for four weeto; ICR end micro bodies (Mb) ere fotriy ebondsnt. but decreeee in HER It not evident Nucleus (N): giutsrsldekyds Oto, fixetion (urenjrl scetete end leed, reduced from x 30,400). Fig 19.--Portion of on hspetoeyte from mouse given toxic rice bren oil for 26 weeks, ilhwtrettng or geereneo of lyoooomoo (HD one wok deveioped Oolgi'e eompleme (Q). Mueleue (N): oomliow floodon (leed, reduced from x 28,440). the livar showed (light enlargement of Kupffer'a oella. Tho livor of a squirrel monkey that received a total of 134 mg of CBP per kilogram in she weeka alao allowed anlargament of Kupffer'a calla containing arythrocyta and dilatation of ainueoidai In two cynomolgua monkey# that were ted about a total of 600 to 600 mg of C8P per kilogram in four to eix week#, notable changes included the appearance of largo vacuoiaa in tha hepatocytaa at tha pariphery of tha lobuiaa and aailaipaiuaait at Kupfhr'a calla. Two other cynomoigua uionhayi wen Arch Snuinm Htolth--Vol 31, Nov ItTO MCJNS 083066 028 CHI.OHOBIPIIRNYL POISONING--NISH/XV,Ml \* V V '.-W. N -<..i--- ...^u'NAi.V-------- N fi9 20.--Are# of on ftepatecyte from mouse prepared # in Flo 19- Mitochondria (M) vary in ai. MlcroDooiaa (Md) are diaperead. Osmium fixation (load, radwoad from x 29.600). given a total of about 300 mg of CBP per kilogram in 20 to 32 weeks. In than cases, many vacuolea ware aaan in enlarged Kupffor's cells* and some hepatocytn contained granular materials and vacuolea (Fig 9). The hepatocytn of the monkey who died after 32 weeks revealed alight fatty degener ation in peripheral lobular areas. Then mi croscopic alterations in the monkey livers resembled thorn in mioe. Electron Microscopy.--Mice Given CBPFree Oil.--In electron microscopy, no ob vious differences won sssn between the liv ers of mioe given olive oil and thoro given nontoxic rice bran oil. After four weeks, alight increero in cytoplasmic lipid droplets were noted, and the abundant mitochondria varied in sin end were spherical to ellipsoi dal in form. The nuclei were round in out line, and many contained nucleoli Other cell organdies such as endoplasmic reticulum, Golgi's complexes, and microbodies were nor mally distributed. Stores of densely stained glycogen varied in amount and localization from cell to cell (Fig 10 and 11). After 13 weeks, the liven showed slight increase in microbodies, as well as lipid droplets. Some of the lipid droplets wars partially or com pletely surrounded by mitochondria. In 22 to 20 weeks, moderately increeled lipid drop lets were noted in meet of the bepstocytcs. Golgi's complexes and SER (smooth-sur faced membranes of the endoplasmic reticu lum) were more abundant and contained electron-opaque substances. Stacks of RER (rough-surfaced mambranae of the endo plasmic reticulum) wore fairly shunrient for the duration of the experiment and, far the most pert, were arranged in parallel (Fig 12). No novel observation were anode with regard to lysocomaa, peribiliary spaces, and nuclear structure. Mice*Given CBP in Olive Oil.--Adminis tration of CBP led to widespread alterations of the ER (endoplasmic reticulum), which persisted for the duration of the experiment Appearance of cytopfaamic industans and increase in microbodks, lyaooomsa, and lipid droplets were also nhasrveri Serial observa tions were as follows: 1. Four weeks after commencement of the experiment, proliferation of the SER was apparent, and individual ciatemae contain ing electron-opaque materials were swollen and veeiculated (Fig 13). The RER were decreased with reorganisation into rough surfaced sinuous cistemae. One to four myelin Aguno were praasnt in about 10% of the hspatocytas (Fig 14 and 15). Slight Incr--a in mfcrobodiaa was obmrred at this period, but lipid droplets ware not yet incr--iri Arch Environ Heollh--Vol 21. Nov 1970 MOMS 083067 CHLOROBtPHENYL POISONING--NISH1ZUM1 m 21.--Ann at an htpHttyM from cywomotgue mewhey fee total of 319 mg ef CSP m 22 wn*i. showing lucre--<1 vsafcWec UK. Nucleus <N); gtatorsMsltyea Oee, ft0a0sa).se <>eee, red--* ttam * M, Plf 32.--Cf-geS MupHer'e ceM of menMey lei total el lea mg ef CiP in 14 week* shewing numeraue veeeetoe wig enetoee el btaed oaH (Mi) Mi cytogtami. reOeeei treat n 12,000). Glycogen u disparted among retaliated or tubular SBR. 2. Alter 13 weeks, the hepatocytea con tained an unusually large population of microbodire (Fig 16) and slightly increased lipid droplets. Mitochondria showed varia tion in sin and form, and lipohadn granules were obeerved in peribiliary arses. Fine gly cogen granules were dispersed among vesicu lar SER. 3. After 17 to 22 weeks, lipid droplets were increased considerably with groups of microbodiea (Fig 17). Mitochondria showed a possible increased variation in aim and shape, and, occasionally, Intiamitochondrial inchtoione were found. Golgi's region was well preserved and appeared active, as judged by accumulation of election-dense products within its membranes. 4. After 26 weeks, marked alterations of ER were sustained. An increase in lipid droplets or fat vacuoles was noted, end many microbodiea ware present does to lipkl drop lets. Mice Given Toxic Rice Bran OiL--After four weeks, alterations of RBR ware not as remarkable aa in mica given CBP in olive oil, but an increase in SBR wee dearly noted (Fig 18). The mitochondria varied in sise. The hepatocytea contained abundant glyoo gen stores, and lipid dropiate were slightly inoeaeed. Arch Envirtm Health--Vol 21, Nov 1970 MUNS 08306 630 CHLOnOBIPHBNYL POISONINO--N/SH/ZUMI After 13 to 17 week*, increase to SER end in hepetocytes by administration of CBP an decrease to RER were remarkable. Most mi generally slight whrei judged by conven tochondria were surrounded by parallel-ar tional light microeoopy, but several marked ranged RER but were almost normal to lixa Alterations in cellular organsItea ware found and shape. Glycogen deposits were preserved by electron microeoopy. fairly well. At 26 weeks, increases in micro An outstanding abnormality of liver crile bodies and appearance of a wall-developed caused by administration of CBP to mice Golgi's complex were revealed to addition to and monkeys is an increase in the SER end tM alterations of ER (Fig 19 and 20). Gly a reduction in the RER throughout the ex cogen granule* were dispersed sparsely. periment It is considered that proliferation In conclusion, the hepatic osll alterations of tha SER to tha hepatocytae estraapende induced by administration of Undo rice bran to hyaline cytoplarenJc toduatons (hyaline oil were essentially very similar to those degeneration) recognised with tha light mi- caused by administration of CBP In olive oil. croeoope and is related to the Uver cell hyper It was considered that ths slight difference trophy. Proliferation of SER in the hepeto in severity of alterations was based upon the cytes is generally seen after treatment with difference in the concentration* of CBP: various toxic substances such as 3'-mathyl-4- about 1,600 ppm to tha toxic rice bran oil dlmtthylamtoomob*nwne,te dfansthytoitio- and about 5,000 ppm to olive oil. samtoe,** thtoacetamid#,1* athkmfce,1* .-na Monkeys Fed CBP^-Ite hepetocytes of phthyl ieothianate," phanofaarbHal aodhan,** monkeys fad CBP revealed tha increase in and chlorophenothane (DDT).** Clearly, SER. with dilated veeidaa (Fig 21). Ths proliferetion of SER cannot ba conidared to mitochondria ware generally decreased in be a specific reaponse toducad by CBP. the density of tha internal matrix and varied Pouts*1 has pointed out that the SER of in eisa. Soma mitochrondria contained to- hepatocytea may ba an important sita of tramitochondriel myelin figures. Lipid drop action of several drugs and poteen*. In fact, lets and microbodies were of normal appeer- it has bean observed that an increase of SER ance, but increase to lyaosomee was slightly in Uver calls is ctesaly araodated with an noted. increase of drug-metabolising ensyme in tha Kupffer's calls slwwed modarate cyto microsomal portion cf Uver homogmatas.**-* plasmic swelling, with dilated ER. In tha A suggestion that than rhengsa reflect adap cytophnm, there were an increased member tive phenomena lor enhanced drug dstoai- of lyeoeomra and many vacuoles, soma cf fication could also be made from CBP which oontainad granular materials (Fig poisoning. frreiasquenUy, it is also fully con 22). In a few places, three wan dtooorrttou- sidered that disturbancaa of lipid metabolism itiee of the odl membrane, suggesting in such as inoreaaa of triglyceride tod in creased fragility and subsequent rupture, blood*4 are toducad as tha result of tha func possibly during rrnreeeing tion of proliferated SER Haas changes of SER ware accompanied by a disorganisation Comment of tha polarised arrays of the RER end by loss of surface ribosomes from ths etructuaUy The oral administration of CBP in oil totact paired membranes. resulted in enlargement of ths liver and Cytoplasmic todustons, which are com vacuolar or fatty degeneration of Uver cells posed of concentric whorte of closely packed to light microscopy. Unas findings were smooth manbrann, were seen in ndos re in accordance with the observations of ceiving CBP. 'Hwee cytoptemfc indusione others.*-*-14 Bennett et al14 pointed out that have been generally referred to se "finger chlorinated biphenyl resulted In liver prints,M*4-1* "nebentan,"*1 or "figures myeli- changes markedly different Iran thoee niquee"34 and have bam observed after treat caused by other well-known toxic agrata such ment with various toxic chemicals.1**1**-** a* carbon tetrachloride and chloroform poi Hafdson at al** suggested that some ensyme soning and thought that this type of injury functions reside in these rytnpterenip indu- waa paraistent with alow raeovaey. In oar aions. Norbeck and Alien** oonsfatered that study, tha morphological altaratfcma induced ths mreribrana arrays would provide a graat- Anh Environ Hoolth--Vol 21, Nov 1970 HONS 003069 CHLOROBIPHENYL POISONING--NlSH/ZUMl 631 ly increased membrane surface area to en* sura maximal contact of the toxic agent with the detoxifying enxymes. Various suggestions have been made as to the origins and func tions of thces inclusions on the basis of the morphological relationships of the formations to other organelles and the overall effects of the inducing agent on the cell. However, many details still remain for later work. Diminution of glycogen stone is another interesting feature in the preeent experiment This is pertly due to the fact that the miee receiving toxic oil consumed approximately 80% of the amount of the diet consumed by the control mice. However, e hypothesis has been proposed to explain tha reciprocal be havior of the SER end glycogen stores.1* Bnini suggested that whan diminution or lorn of glycogen and permeation of glycogen areas by tha SER occur almost simultaneous ly, circumscribed acidophilic anas might be seen.** As for our findings in light micro scopy, appearance of acidophilic granular materiirfs in the cytoplasm early end of vacuoles or hyalins cytoplasmic inclusions later might be the results of hypertrophy of SER end diminution of glycogen stains. Four months after the administration of CBP in olive od was stopped, an incraaaa of vesicular ER, with dilated sinuous oistsmes, was still ob served, but in tha suns period, no alterations of ER were noted in the hspetocytee of mice given the toxic rice bran off. It wee suggested that theaa alterations of ER an .adaptive phentmena to CBP and pesatet in proportion to the concentrations of administered CBP. The microbodiee in experimental mourn liv er slightly increased in number early, but these organellas were markedly Increased af ter 19 weeks. It has been reported that the microbodisn take part in lipid matabotiem and glyconeogeneeie and that Imrsase in number accompanies changes of their struc ture in liver injury.** In thie experiment, en increase in microbodiee might be recog nised as the morphological expra--ion of metabolic changes induced by CBP. In control mica, fat wee abundantly ob served as cytoplasmic globuiee from an early period of the experiment whereas in experi mental mice, increase in lipid droplets wee observed after 20 weeks. 'Rtsee findings sug gest that altered metabolism tortured by CBP prevents an accumulation of ttpU in the early period of the satpsriimnt end in sults In- en increase of neutral fat to the blood. In the monkey, an incnaae to tipfal bodies wee noted in Kupffur'S calte of the liver injured by CBP, but hepafeoesttutor H|> id droplets weie not increased. This tod does not resolve the question of whether thto dis parity is due to the difference to animals or other factors. For enanpis, monkeys were fed CBP not disrobed to otto while miee received it in oils. In our laboratory, other work is going on to clarify the differences between theee two caara. In the near future, more refiAed combtoations of pathological and chemical analysis may elucidate its mechanism of action. 1. KhmInm M, at al: An apUmMaiii atudy on Yuahn. Fukmoka Ade Mad ShSIt Bit. IBS* 2. Jm JW, Aldan Hft An aana farm daaiaalai forts Anh Derm Sypk ttttttMOM, 1A 3. PiwdasUI V: DrtTaona HilsHre. Mad Imoro 4li131-146, 1964. 4. Mrtgs JK, Atom JJ. Kartte BL: CMoraene fraaa an imaaual oxpeaura to Aiaalar. JAMA 16411417-1416, 1964. 6. flahwntte Li DamaStii from aynthotla raafan and waxaa. Amor J Patels tfaaJM StdWS 8S1 1666. 6. Drtokor CK, Wanes MV, Daanatt OA: Ita li98S-SlL 1SST. T. MUlsr JW: fie ffanM Hap giUteHN, 1944. & Bammtt HB, Loll frCbUdtea aa a Mate far hulterfaf Santfvaa. J BMn Btnhmm Cytmi kill. 114,199a a. Minwrtg Oi hrtai afcaaavattena an a ySaa Ms Filth fnfarntftewrt Oawraaa an fksSw Mleroaaspy. Phitodotphim. 1963. New Yad, i Pleas Ine, 1963, p P4. 10. Lull JH: Ii^rni iiamla ia *1400414,19SL 1L Trump BP. Bmurklar BA, BanffIt BP: A mated ter apaay sssttaro tee H6N ndciew espy. J VUrutmet Has M43 343, 1BSL 13. Watesn ML: Wtetotog of tens aastfsro far alaateon sakNaeepy Hb baavy naataia. J Btepdyi Btastem Cytoi 4*76476, USB IS. MUtsnlf A madtSad paeaadaae tas had statatof af tten seeUaae. J Bfapdya Bfasdam CySal 11I79S-7S9, 19SL Arch Environ Health - Vol 21, Nov 1910 MONS 083070 CHLOROBIPHENYL POISONING--NISHIZVMI 14. RooneU GA, Drinker CK, Warren MF: Morphotogkai chenfee in the liven of nto moulting from eipoeure to certain chlorinated hydroeeibcna. J Indumtr Hyg Toxicol S7-198, 10*8. 15. Porter KR, Brunt C: An electron mkroecope tudy el the early effect* el S*-MeDAB on ret liver cells. Cancer ffee 1*1*97-1006, 1000. 16. Emmelot P, Benedettt EL: Changes in the Ane etructun of rat liver eella brought about by dimethvInUroaamine. J Biophyo BiocHcm Cytol 7t3m-3oa. loan. 17. Tboenee W, Bnnnenrh P: Blektionen- und lirhtnihioehopieche Untersuchungen am Cytoplee ma der LobemeHen nach akuter und chronlecher Hitoaeetemid'VerfUtunf. Virchow Arch Poth Amt SUiSBMBS. 1002. 1& Kerman L. Eber L, Fltegoreld PJ: liver ceU depeneiBlieii with eUiienine ertiidnletiatiwii. in Proctedfitgs of the Fifth Intarmtioml Congrom on Electron Mierociopy, Philoddphim, IMS. New York, Academic Prate Inc. 1962, p V-6. 23. Renuner H. chancee in the liver t 140t1657*1660,1060. 24. Usawa H, at al: Hypwglyceridemfc meat* from intake of rice oil muiaarinafd with rbleibe ed biphenyls. Fukuoka Acta NriMOthtM MB 25. Emmelot P. Benedetti EL: Scam ohomoodn ture end function oI the endeptomde ntahn oi ret liver cells, in Harrie RJC (ed): Protein Bimogm Ikeck. New York. Academic Pieaa Inc. 1001. p Ml 38. Houilter C, Simen G: Centributlen da In mtcreecopie llectrenique eu pregrti da aae earn neiaaancca an cytelegis at an hiatmpathalefie hkpm tique. Rev Int Nepal 18067-80* 1062. 27. Hardson PB, KaHmbarh IP-. Blectien mime cope etmllee m ensymo aetivity and the ieeialtew al thiehydantein-indmed mycUa Hgutee hi ml Mean J Cell Bid Mi4M IM. 1065. 28. Hruben Z, Swift R WUc RW: Early el the cytepbnm el pereechymel liver celle in aIphe-naphtfaylieotfciocyaiMte.induced cirrhosis. Lek Invoot 12i765>700i 1068. 20. Kanban PB. Gervki PJ, Jemdis RB: Ftoe eliuctuml Aengee In mt liver Induced by pheneber bitaL Uk /avast UiMO-lMT,1184. 9L Ortega Pt Light and Elactren Microscopy el DkhiorodlphepyitrichbireWfne (DDT) pnienateg In the rat ll*ar. let fowd IMRAN. 1066. 28. Foute JR: Inbtactkm oi druse and hepatic rnkwitatt, Pod Proe 21:1107-1111, 1081 bata^thbnylaianhw fed min Fed Prm 8BH8B 1081. /avert 908-916.1060L 81. Popper R Miftcr F: changes oi the liver. Ann /aMm Mod I 1081 ULTRASONIC COMMf UNICATION Tlia uppar frequency limit ci human haaurinf ie geaarally about 17-20 kHs in young peopla and, although for technical rearm audiometry it ealdwn centtnnad above 12 kHs, it b fairly amp to show that tbo Umit tonda to decUno with aga ae an npreoricn oi prwbycaeia. It has bean Habited, however (Pumphrey, 1900), that tbo HmttnHen ia entirely due to oonductien loai and that eounde praaantad at riflhlrnt intensity by bona conduction are audible, without further vernation oi pitch chanf* to more than 100 kHs. It appear* that the human cochlea la capable of raapandlnt to Ugh "uUnaaMa" frequencies but doaa not normally axparianoo them and ia unable to perform fnnnamij analyaia upon them. Time limitations of the human ear are not ehared by many anallor w-mmI*, for there la good evidanoo that many apedea not only hoar and analyse nltt--mind. but use theee frequandao for communication. Balls <1987) haa shown that mica and Poromyogua show poatorior coUkular reepeneea to at least 100 kHs, with greatest enadtlvity in the range 10-80 kHs, Ndrot (1980) found that baby mica aanmunieate with thrtv mother at 60-90 Ufa. Sewell (1987) haa found similar behavior in ten other iperim of Myomorph rodents and shown that the adults of some apodaa Indicate agiiemin and mbmimivo Intenticne by ultrasonic signals, The hearing of bate is especially interneting, because the Micrachiroptem <how a highly developed ferae of ultrasonic echotocation.--Pya. J.D.: "Hearing hi Bats " in da Reudc, A.V.S., and Knight, J. (eda.): Hearing Mochaniomo in Vrrtebrefer. Beaten: Little, Brown * Ca, 1988, p 68. Arch Environ Hoatth Vol 21, Nov 197i HONS 083071