Document aLaeyJqZ52Vg1vZ9Ojm1nemM

7 M K I U A T I O X O F A S M K S T O S F 1 B K F S H ; u \ | S I T , ( 'I ' T A X F O F S I X . J K C T I O X Sl i'KS iX ,\1!CF K . K A N A Z A W A . M. S ( '. HI I? D E C K . It I, (' \ P T K P \ \ i > F. ,1. V. RON F r o m Ua < ' h i a h r / t a l l y R t n a u t h I list it i i t r . I tr - l t l ilt itf ( ' ana i Rcur ar vh: Royal ( ' i m a r I h a f i t l a l , Fi i l / uun Road /.o/e/oa. >S'. II'.,'! (*(l for jmMicai n>n NovciiiIht I U l ih/1> -S U M M A R Y .-- C ro e id o lite asbestos fibres, suspended in p h ysio lo g ica l saline, w e r e in je c t e d s u b c u ta n e o u s l y i n t o one o r b o th f l a n k s nf 05 (111 \ la ic fe m a le m i c e ; 75 c o n tr o l m ic e received in je c tio n s o f s a lin e only M ost a n im a ls w e re k ille d at chosen in te rva ls of between 2 and 42 days afa r injection but some w ere left for longer periods of up to bid days. A t autopsy , many l\ m plioid and n o n -lym p h o id s tru c tu re s were re m o u 'il and exam ined fo r the p ie s riu e of asbestos by the followin' techniques: haem atoxvlin and eosin stannic; followed by conventional and polarized Ik;lit microscopy ; P erl's stain : m ici oincineration followed by phase-contrast m icroscopy; m aceration w ith IvOH followed by phase-contrast m icroscopy; and electron microscopy. A c o m b in a tio n of h a e m a to x y lin and eosin si im in t; and m ic r o in c in e r a tio n was found to be the m ost c o in c id e n t and re lia b le m ethod fo r d e m o n s tra tin g asbestos fibres in the tissues. E lectron micros* opy was essential fo r detecting very s m a ll fibres and fo r lo ca tin g th e m to spet ific in tra c e llu la r stru ctu re s. The m orphological findings indicate that some m ig ra tio n of asbestos fibres aw ay fro m the in itia l site of inje ctio n takes place D isse m in a tio n is u su a lly along ly m p h a tic p a thw ays and fibres tend to a ccum ulate in the ly m p h o id tissues, p a rtic u la rly in the regional (a x illa ry ) lym p h tuules; sm a lle r am ounts were fo u n d in in g u in a l, m e d ia s tin a l and lu m b a r nodes. I he fibres w ere u s u a lly in tra ce llu la r, lying inside the phagosomes of nun inphages, but la rg e r fibres were so m e tim e s encountered ly in g free. S m a ll n u m b e rs of fibres w ere seen in th e spleen and also in n o n - l y m p h o i d o rg a n s such as the liv e r , k id n e y s an d b r a in suggesting that some asbestos may enter the blood s tie a m . T h e re w as no evidence of massive or selective spread to subseioxal tissues in the thorax or abdom en, though tra p p in g ol asbestos fibres was obseived in p le u ra l " m ilk y spots " in l o n g - t e r m surv iv ors. The possible t ole ol m ilk y spots in the d e v e lo p m e n t of p le u ra l plaques and m e so th e lio m a tn is discussed. T i l i a t K is m cre a sin g e\ donee I lia f e x p o s u re In a.--In .1 os is associated w if li (lie d e v e lo p m e n t of m e-ol h e lio m a f a in man ( W a g n e r M '-gg- and .M arehand. I '.Mitt; W a g n e r. I'.Mia. X e w h o u - e and T h o m p s o n 11ni.b N - h k o ll < litirg a n d H a m m o n d . Ihfib). T h is a s s o iia lio n is borne out b\ e x p e rin io u ia l studies in an im a ls where nu'sot h e lin m a l a h ave been in d m ed l>\ n j e i i i u g hi `M i o i hreet I\ inf o ( In 1 p le u ra l space ol ra I s (W a g n e r . I'Hig). h a n is ie rs ( S m ith M i ll e r ( ' l u n g and Solihull'. I'.Mio: Sm it h . M ille r. K ls a s s e r a n d l l u b e i t I !ll>.'>) and fo u ! - *Peat nek a n d Peacock. PMia). Poe. C a r t e r W a lto i,- and l l a r m g l o n (P.MiT) re p o ile d th e u id u e t io n o f p le iu a l and p e rito n e a ! mo,sot h e lin m a l a in mice a l t e r a -b e g to - had been in je c te d s u b cutaneously into the Hanks n . 'in itia lly remote Irom the regions when' mesot lu'liom at a subsequently developed. At th a t tim e, it was suggested th a t m i c i : a r i o x n r a n i k s t >s i ' i i ; i : i <17 asbestos fibres miulil spread widely from 1he oriion.d m |ecl ion sit os and find t heir wav in a som i.-elei 11v<' I.i'I iit 111 lu I i11' mi Ii.si'i'i ma I la \ cl' ol t lie l !Kira ci c a nd a Imh mi inal v iM'cra. An at t empt has now liccn made to exami ne m detail the di-seminat ion ol asbestos lihres altei their in |eel ion into the mi I k nt a neons tissues of mice. A preroipiisito ol an v s n i i st udv is a careful e\ a hint ion of t he 1cell nil (lies available for detecting asbestos in the tissues, and the <ui i ,111 lesiilts are aecordimrlv presented m two parts first. an appraisal ol si\ mi (hods lor ident ifviiif: asbestos fibres in the tissues, followed b\ an account ol the auatonneal distribution ol these fibres a l t e r subcutaneous injection into one or lint h Hanks o f mice. A siteuton .M.vnuii vi.s \ m > m icrnons A I n I r e f e r e n c e sample o f eroeidolite w as supplied bv Dr. d . S . Harinji ton (South Afri can I nsti tute tor Medical Research) t he distributions of the lilire length were determined b\ Dr. \'. Ti ml i r el l (M 1! . ( ` I' neimioconiosis Research l ' n i t . Dlaiidoueh Hospital. I 'enarth. ( l l a n i o r o i n ) \ p p r n \ u i i a t o l v AU` > of the fibres were between 11-2 and 1-2 p ill length. and the rema mi ne fiO*',, between 1-2 and 2-0 f t , less than .V',, exceeded 7 t in length, ' f he tihres were suspended in pli\ sioloiiical saline before injection. Mire One hundred and seventy OliA/' Dae female ni ne were used, afil'd (>--S weeks and weifrhiim about 2 a e. T h e y were mai nt ained on a cubed diet (Diet No. N(i, Messrs. Dixon Ltd.. War e , Herts. ) and wa t e r ail l i b i t u m . and were kept in metal cnees. 10 mice in each. ! 'o m i ne t o f ext eri t to i/t 'flic mice were divi ded into -I groups. Detail - ol treatment are shown in T a bl e I. Most ol the animals were killed at i Ih m 'ii intervals between 2 and 12 days after injection. Some mice were allowed then lull life span and survived lor periods ol up to Is dav s (( roup I ) and (>2a dav s (( roup 11). T a h i .e I.,\u nf ( lI 1)1111 miir L 10 11 ;>/> 11 1 .'0 l\ ! >t t a i l s o f T r e a t m e n t i n T i n t a n t i C t t n l r o l G r o u p s '[ Mitili 111 :tn me i 1. i<1f>i111' 0 1 ml |i i\ '_r 11mi 1111* i 111, i 11A 11111tut ni'l v 11 ' t 1 llllvs iill sllfCI'sslVI tIji\ 1e hi , 1 i li hi c 1* 1 ml |>1\ 1' !< A1' 11 -,0ll|i' I li |, , I d !1"`HfHtin Rid , 111 " il 11.i 1Lk " 111\ . " M 1 (1:t\ . 'M i n i ( h l"l<Mt< fl -illlllr ri1\ . l'i\ n ,i m <2 mu|> 1 il 1 ml | ft 1>1<a'lftt 1-it ll Ik nip . _i\ <'ii ti-s in j.i-1ji11>11. Auh v ! Wice in en ,ii| 1 tinti 11i i n e ,1,-m I 11"111 ,t -mul< 1,11< ) 1 t,111 m ;t <1 11 tm i l it n i ) 1" 1 h e w M IICS IT! C l o l l ] - 1 a n d 1 \ i ............. 1 IV < ' 1 r m i t - ` j .11 .i t v' dal < li. al-" .1I l i H M tft I i iindttinK hr c n n i|i2 \ C'MUCC in " d i r w t t - l l - c i l lil ; II tit |C( 1M ill s \ n i m a l s wer e k i l l e d by e t h e r v a p o u r o r bv c e r v i c a l d i s l o c a t i o n . S t a n d a r d necropsies were pe rfo im e d . Inje c tio n sites were e x a m in e d for evidence of ' )S K K \ .\ \ / . \ \ \ \ M s I . 1 : 1 1: 1:1 ( K . l;. I c \ I, I M : \ M I K .1. r . !U i h i n:n i \ erl ci 11 m | c i l i u i i ni ,i I m 111' i n i n t h e 11 [, ,t, i , i i ,i ! mi i I m r . i c i c i . i v i t i e s . C . i i v W.1.-. I <1k('l I I II .1 VI (III I I I" ` |'l I II.I I IM|. ' .1 M i j I I I - M i l l I I I ' -I II - dlM'lll'J .! lit M| M\ . T i l l ' Ic>llo\\ mil: tissues u r n 11*111m \ i 11 H i m ..... A !m i \ ,, i>m:m ! mi i axillapv n m n in .il MIC! I 1,1''I 11III I I Mill I Ml ,1III I Mil ,1 `I I ! Cl IC I \ !II | ill I IMl 11 , | !| \ III I I ' I I I I IlC, I ,1III I I I PSopll.le IIs in i ii ic I ili ii'k 111 ,i i'l ,iiii I ! n 11 111 i m i 1 111mi k 11\ ci 1>h i ii !j i 11ic\ ,s o \ ,u ics . hr; i in . east i'i mi l (Ml m , 11 t r a c t t h e 111m i ,11 ir w a l l . mi l .i I n l o i :, 11 i.i I w a l l . I m t h nl ;m<l a w a y I IMII I I IlC 1I jeel II III s|| r ( s , M r l h m l s //.vi r/ / hi ili l< cl i mi H'-ht s/ic I' l ' i i > m lh' lis.sin < U i / h l hi irni.^riijii/ follow s : T h e t i ssues I i \ e <l in l i n i i i n s m i I i i I i mi i w e r e ill vest I ya I ("<I ns I I 'll ni flill s e c t i o n s | i r e | Mi ' ( ( l nt m ii w e r e s t , l i n e d eil In r w it Ii l i n e i n a t o \ y t i n a n d covi n o r l i \ I '<i l > i n e 11m n I l o r i ron a n d e x a m i n e d h \ c o n v e n t i o n a l 1i lli 11. in iero.-eo| i\ 2 . T h e l i a c m a t <> \ \ l i n a n d e o - m s t a i n e d s e c t i o n s w e r e al so e x a m i n e d u n d e r | i o l a r i / e d 11e 111 l oi t h e |>i'esenec ol l i u ' e l i i n m ill | >a I t a les III t h e tissues. A It ei dew a \ i lea w i t h x \ lo| a nd a l c o h o l ,n Id 11an ml v , l i m n w ere i n e i n e r a t ed at .Vlil ( ' . l o r I l i o n i 1 i n ,i S I ' W It m i n oi i a uea a I o r (< l a l l e n k a m | i ) . T h e y were I lieu d r \ m o u n t e d and e x a m in e d h\ |iha-e i n n t r . i ' t nueroseo| iy. I'lh rh m i i i i / o i i M i i y / Tissues w e i e lixed in e I u! a ra I d e h \ ( le in I , a M e a e o d v l a t e l u i li e r ( | d l 7-1) l or 2 hor ns p o s t h x e d w i t h I '',, o s m i u m t e t r o x i d e in \ e m n a I ai et a I e I u 111e r ( | d I 7 - I ) l or 2,0 m m ut i s del i \ d ra I ed \\ 11h m e t h a n o l t r eal ed with |>ro|>\lene oxide and then e m hedded in a ra ld ile I l l i a tl n n section (lion to w e r e ( i l l on a 1111 \ l e v - t v | ic nil ra m i e m t n i n e S e c t i o n s w e r e s t a i n e d w i t h K a r n o \ s k \ ' s alkaline lead stainin': m ed ium and e x am ined in a Siemens or a I ' ll ili I Is K M at III elei t m i l in i rl cm o p e Sa III pies o| r i m HIc.111e I i hres w er e ell) I ledcled in a i a Id it e in t h e sa I ne w a \ lot cmi it m l pu r poses Tis si n iinin i a h i m . T h e l es i du e ol each s p a m i n i was m i n c e d w i l l ) a line scalpel (a new M a d e l ienm used loi each s pe c i me n l o a v o i d I r a i i s l e r o f asbestos I ro m o n e t i ssue t o t h e n e x t |. a n d t h e n mac e i a t ed h \ a m m 111iea I ion o f I lie1 me t hod d e s e n li ed b \ h o l d ( I u<i7 i \ l i mi t m a c ol I he mmc i cl t m u e u as m i x e d w it h 10 ml . ol 10'h, acpleolls p o t a ss i u m h v d m x i d e a n d l i e i t c d on a w a t e i h a t h loi I 2 hours 1111111 t h e tissue had been l i ne U l i a a m e n l e d T i m i c s i i l t i m suspension was them ccut ri f ueeel w a s h e d w i t h l l i m e i b a i l e e s ol d i s t i l l e d w a t e r a n d a s m e a r w a s prepared Irom the deposit This was mounted drv lor ex a m ina tion liv pliase- eontrasl m lemseopv i; i > i i 1'^ Kar t 1. ( ' ( I H I ( H I 1 I s o i l d \ 1 I I I / d l l ' - l/c I'll tl I- l i n t - ' >II i t d T i - II" f h h i r l i i m i i ) . 1- si l l s i - i s T i n a pi (`' (' I I f e ol lib n - m 11( s i j n ||( n. Mi l s 1I-SMi's . n i l ic in |ec! ion si t e ill 'lest m ice i: i \ ('ll llsln `- I n s , i ud t im ,,l i - e m e ol sin h l i l a c - Ill t Ml I 1m l m i c e i n |(`el ed V n h sal 1IK* p m \ a h `i 1 t h e | >i r i \ < a ml m "aa 1i \ d ( l a 111 <ih 11 . ii \ l or assess me t h e ' a ! l i r <ol d i l l e t e lit met 1MMU In 1 ili I n l i l r j ,t | m*-1 O' i'l.M m \ . m o i l s l i ' s l l C s . ' hi m i n , ' ' i l l <i'I i scM a 1loi I - Hi m I n ! l nil ! 11 1he - i \ i,n 1hoi I s Ms ( d a i d v i i m m . i r i / r d mi T a b l e I I . ( ' c r t a m Leami al j n mt - .11 (' \\ Ml tl i - I n - ms d II / i u n i [Ii ii s f c l K i l l h m i r H i " i i i ' i ' , m i l l <t ) s / ,, 1l i / h l i s m >d < i d f u f . The ranee o k asl ><^l< IS III Ill's M "( "It i> o l | \ l ( l l | s | \ h m it ed i \ l :h e i r size . 11m* I m<*1 hod i s i i n s m t i d i l e I 1111 K. k \ . \ \ Z \ \ V \ . M S. ('. l i l l i l '. K C k . 1!. I.. ( ` VltTKI! A M I F .1. ('. KOK l o r l i b r as nf less I h a n a hi m l 2 u m l e nc t h. 1l a e m o s i d c r i u ar a m i l l ' M i l l cn a n n m u l al e ill cl< isc f( 1;11j<>11 t n asl a ,-l i is l i l u e s a n d p r o v i d e ,1 Useful i n u i k e r in l e r t n i n t i ssues s u c h as I\ 1111111 nodes. T i m ,i -`m l a t i m i n m i l l o n n d m ^ u l i d | i n r i m i h \ i n . i t o i l s <m f a n s a n d t h e d i `l m l m m ul s m a l l l i br as m l i a c i n a 111 \ \ tin a n d c i m i i s t a i n e d malarial from such liaMim isdillnaill a / / /ill r i i j j i li . s i t i u n i s - 1`i r l s s h u n , i n j h l in irrn.sri i/ii/. \ 1111<)n LT11 a s h m l t i s (liras ar a i if 1mi a s s o c i a t e d w i t h h a a t n m i d a i in. a n d a r c t h e m s e l v e s a o a l a d w i t h i r o n c o n t a i n i n y p r o t e i n s . s l a n t s l o r i r o n a i a no t | >art ian la i I v 11a 111In 1. A d i s l i n a l m n r a i l ha d r a w n hat wean Ilia d a r k e r s t a i m n y a s h e s l m Idiras and paler s t a m i n a l i a e m o s i d e r h i , l i n t in o n r a \ | i a r i a n a a I l i a ma t h o d is m s a i i s i l i v a as lass I l i a n a l l 1' ,, o f eroeidolite lihras stained positively lor iron. a // / m i n(in si l i m n s m tr u u n n m i ul k m ; ji/iiisi k 'di i I ni si in i rrnsro/ n/. T h i s mat h o d appears to ha m ore s a m i t i v a th a n the e x a m i n a t i o n of h a e m a t o x y l m and aosin st a i n a i l m a t a r i a I . It is p a r t i<n la rl \ 11-af'l 11 f o r s o l i d 1issues snai l as li \ c r a n d l i r a in. K n i t l i c r m n r e . it is st ill possible t o locali ze li hras w i t h i n i n t a c t t issue sections. K r a y m e n t e d c o l l a y e n m s v . how e\ <t yi v a rise t o false posp i \ a r e s u l t s a n d t he r e is a i isk ol contamin .it ion w ith extraneous asbestos p a r lic n lirlv Irom the linin y o f the m ia roin cin aral or oven itsidl. ( 'a re fill f o c ii 'i n y m a v ladp to disl mulish contam in atin u asbestos lihias if the\ lie ill a more siiparlicial plane from the main specimen. M n c i i n l i i i n . /i/iu.st - n n i t m. s l in iri ' i i sr n/ n/ . T h i s p i o i c d n r c is m o r e s e n s i t i v e t h a n h a e m a to x y lm and eosm stainiiiy. lmt it carries a serious risk o f false-positive r esult s, par t i c u la i lx in r e l a t i o n t o s m a l l fibres. T h e r esul ts are i m p r o v e d if t h e macerated tissues are nncroincinerated before examination. u ii in i n [fi n .si r l i n n s . I nn i i i nh i.ri/l i it m i l l t o s m / i i i h u r . i i i Hi /hl . - A l t h o u g h us e f ul f o r d e t e c t i na l a r ee l i h r a s m o r e t h a n 1U / / i n lei eat h t hi s I ee hn i c p i e is u n s at i s f a e t o r v l o r t h i n Ii liras in w h i c h hi re I n i l nance is t o o w e a k t o lie det eel ed. hhhrhiiii niirriisrii/ii/. T his procedure is not suita ble lor serceniny for the presence o f asbestos lihras hut it is in \ .tillable lo r con Iin n m y t lit' presence of, a n d i d e n ti fy i n g , libras fou nd b\ o th e r m e tho ds ( Ida I). It is assent ial for detect iny \ c r y s m i l 11 I i l i ras a n d pa rt i all's .is s m a l l as fid 2.1 A h a v e been idel it tied w i t h t h e elect ron microscope in the present work. Klectron mieroscopv is also required for th(> p r ec i s e l o c a l i / a t i o n o f as b es t os w i t h i n cel ls ( K i y . I). I t u r i n y c ut t i n y . t h e m ic ro to m e k n ife often draas asbestos lihras t h ro u a h the tissues. y i\ iny risi' to e h a r a e t er i st ie t ear s w h i c h a re so met l i nes hel pi ul in In, at in a a sliest os in a s p e c i m e n . 'flic distinction b c l v c u i asbestos libras and iesidii.il bodies in phayosomes or y r a i l tiles o f h a e m , i si dei in oi mel .i m u . is us i i a l l v ~t i a i a l d o r w ar i l . It was con clu ded that the most e l le c tiv e mi I leal lot screenina tissues for a si lest os is a c o m I ana 1i on o I h a e m a l ox v 11n a n d eosi n s| a m m y a m l m i c r o i n c i n c r a t i on. ' f i l l ' ef li ciencv ol the.se m e t h o d s is i l l u s t r a t e d in ' f a b l e I I I . w h i c h r ecor ds t h e d i s t r i b u t i o n of a s b e s t os l i h r a s m 10.1 0 1 1 .1! a n d m n i e d i s t a n t I v m p h no de s 2 t o 12 davs a l t e r subcutaneo us ni|c c ti on ol asbestos in to the Hank, yrou ps I and II are t est a n i m a l s 1n |ect ed w it h a.sliest os ai m i p s I I I a n d l \ a 1 con t rol a 111 m a l s in |ect ed w It h sa l i ne (see ' I a I )|c I 1. d h rcc poi 11 ( s e m c i ; r . I '1 ed n t a III v . I he h in best v laid o f posi t iv e 1 cst ilt s was o b t a i n e d 111 y r o u p 1 in 11111 e w l i r h rceei v ed t h e lai y e r t o t a l d i s c of asbestos In bot h test y r o u p s the p r o p o r t i o n ol po s it iv e results declined in t he m o r e d i s t a n t Iv m p l i n i d t i s s u e. T h e t e n d e n e v I 01 m u r o i n c u i e r a t i on t o p r o d u c e l u l s c - p o si t i v e i c s u l t s is i l l u s t r a t e d bv t h e oc c a si o n a l f m d i u y s in a x i l l a r v nodes f r o m c ont rol ill ice ( y r o u ps I I I a n d I \ ) i nject ed w i t h sal i ne on I \ . T \ i i 1. i: 1 1 1 . / h I r c i i m i n i C i 1u r i ! I n l t l t . 1f i n - h i . ' : 1 l i / , i ) l l t / i l l u n i 7 ' / aai n-. v. ( ' o i / r n r i h n i f f l i . ) f*!f(1/ I h . i n I I - I n / U n < w n h i . i i / l i n n m / F i 1X1 I I S l m u I I K J < / / . <( n in i .1/i n o - / U fi in i n i n n i { M I ) T r -! . m i n i . i l - f II " " I 1 1 :h 11r.r a n i , , l i t ' 0 1 m i -aliti - r lilla leali llankun !{ - a i r c r - - i \ < i l n \ - I I A l i ', ,ii ( 'l 1u- 1 \ MI \ m II.ii \ lai. I J' - M al 1HI -1 1II al ......I r - Umili Mr IH Hir - 1n a u in a ! huiir,- li II , Ili i:t 1 11 It Ij n 1f O l oup i r HI Il l Lf r r o n n l o l i t r /li 1 m i . - a l n i ' r * r . m t n ol ii ' f l a n U <ai 1 li v <m l \ i *t >ut rul1 u n i t i m i - ; Or III 0 1\ m i s i i m i 1 I MI lls III J.rl'i MI | > . I I . & l { ai / '\ Dia { I I A l > (III r \ 11iI I.- . i \ MI .MI Hi ;ts : t. U it ti j ;s . : JS . is .t 1 (I . o i:t . s u .0 a .n ti \i i 0 11 u n il, nlip IV I l A- l ML o- 1 ini - t i l t l i r priviimi n- m r i ' >U|> . II t '( 111 / \ ti 11 0 H _\s j i IH n Dia. { ti <i u ti `i ti <) 0 | )f r - r l i r e nf 1 - l i r - f o-, r i m f i m i r i 1; | UV>< a i r i 1 o f i r a h r - t u - n< it r u l l i l i li t r i 1 Di'- (11-(m<lani ; - < -uh< ut nriruii- T< il al e Miri mi lanr<` ( ) ) l u t a i i i i i i i i I hm ]<M) :un :tt!i ` Min s7-l, ('un. ri M ir u r i l a ti 1 ; P a r i I I . D i m i i mi uni n f . I s/n ^l<)\ F i l m a l o l l n i n mj Si i b i ' l i l i i i ut ni n I ii)ii Inni mio I/o Flniilm T h e I s m k 's in u h i r h a - b e a t o - l i br es m a y I * I n c a l i / r d u i l l h e e o n s i d e r e d u n d e r t i m e r i na i l i h c a d i n u a I he l \ 111( >h<nd s \ a l e n i me s o t li cl i al I i ~ 1 1 a n d ut b e r o r y a n s . it m ust. h o \ \ e \ r r he atiraard Iha t asbesto- nui\ tic depnsited in ad vert en t ly ili d i s t a i l i t issile-, as a r e - ul t n f t a u l t \ i i i | e c t i n n t ec l i n i i | i i e a p r o b l e m u h i r h is p a l i i<111a id \ I i k e I \ t <> nei n r u l i en l e p c a t e d i n j r e t ii m - i >1 l a r e c v o l u m e s oh m a t e r i a ] ai'e II sei I. I ) e - p i t e t h i a re I a kei i t u a \ ( a d pel l et rat i m r t I le tul i t li i ekness <>f t he l i n d \ u a l l . a e e i d e n t . d n i t r a p e i i t c m e . i l n i |eet a >n n| a - b c s l n s nei i i r r e d in a l e u n i s t . me e s . l \ li leal ee ni pene) rat i mi m i n u m i m e Hi m i n 11 <au <j re ai p I w luci n e r e ki II ed he I n r e 111<` e I si 11 d a \ a m i l i 1 .~> ni n r 11 o m r i m i p I I k i l l ed I !( ire t h e ( i -. ' l i d d a m D e , inai t s n f a -i ie.-t na u r i e ari a i aa 1,1li e 11 u l u l e i mi 111Ics n r p i a ( p i r a oli t li(` sei nani su i faci ' in I I a - e c iaes. m eli i i i ne mn a l i n m mi ai I \ 111 11 ic s i i h p h r e n ie spaees, I 1111 pn r t a la | m t i - . i m i .n ni i m I t he a pi cei i d i i me 11i i n ai m i I li f a m i i - u ere anni et i nies In i m i t ] i ri 1ill I mi i t I l a a 1mk an m a l i a \ il \ Si n i a n i m a l a | a \ e he c u e \ c l ud e 1 I r ol l i i li ri h e r di t a ileiI n a i- ii k i aI a ai in 1he aeri ii m - 1li.it I n i I n u . \ n U I H ' \ p e i t e d e h a i m e a u r i e a m i ,U ( h e - uhi u/n m an.-, i n j i f t n m s i l i * , a p p e a r a m ea u er e ai m i l,i i in in me I n i ni r m u p - I a n d I I i are T a 1ile I ) X il ni e r m la a al test i is I l ia ca u r e e p re -e nt in t h aulirli! meni la t i - m c a il all staees ut t h e x p e r m i e i i t . T h e p r n p n r t i mi n f l i l i i ' es l y i i i y i nal i l e t h p h a y n a o n i e a n f i n a i a n p h . i p e a i n i a eased u it li K in r Uni SIM : It.STK i.\ (IF SSI' .F.STos Fl I; K l v I (IT) I l l u s i o n 1)1 t i l l ' I I I 11 I 11i r k 1l("ss (11 ti l l - b, id S Wa l l . W i l l i Ml ! I-I i | III 'll I I l '| I 1r ( l \ All a/..I W,1 . mi | ii i M i -nI mm I r \ | )< i m i c n l --1 \ n . m b r - i o , i | i | i ` ` i i i i | in I im 111 ,i mill'.' . i \ i l l , u \ l \ m | i l i 111H11 u n i 11 I I n I 11 11 1 1.1 \ <i 111 i 111 |ri I n i i i I I x i l,i s i i l i 111l" 1! \ i ,ni( I it w ,is cli',-ir l l i . i t .i 1.1 ri c | ii 111 ii ii t ii in nl 111111 ' 11.111 i n - n <11\ i 11 11<I c,i i i ! i l l s i n l lie 11 in m n. 11 I \ m pli nodes. \ s b r s t o s lilui'-. I,iter cii usi i ! the m n l Inn- ii"l . i r o b - r i \ rd hi I lie i hi I i';i la thi ,i I i n n u m a l a m i I m n l u i n o d e s s i t e s u l n i ! : a n - m. l m \ o l \ H ( l m i n t a c t a 11 i mi l la \ lew l i b i r s ri *. i <-h d i s t a n t nun I \ n i j >11<>111 u r n . i n - a n d ' i - - i n ' s j u i >I >a I >1\ in f I k " M i ii id T l ic | ii >i lit ( i j ( 1111 \ nl I! IM i s i n 111 t In \ a s< 11 lai s11 hi ' ' in k in >w n , i I It HU lie n t l ie sn I lent a nci n a | is m e s a I 11ic sii c i ifm |cct u a i 11m i n i c1r. i in mu I \ in | ill n od e s w h e r e t h e r e a r c a n m n l i e i nl | i u t c n l i a l I \ in | ilia t h 11\ c m m - i ni ncct a nis ( P r e s s m a n a n d S i m n n . I '.Hi I . M n it 11 Ihli.h) n r / in t lie t h i i i . i c i c d u e l It is | i rnlia I ile t hat llinst. aslic.stns fibres t r . u e l i nside i i i .k rn|ilnnjes t l i oi i nh m i i i i h l a m e r lilires mas lie free ill t h e I \ in | ill n r b l o n d . t e n In rite ti l ue s w ei e net a s i n n a 11\ seen in t he ( issues w h i c h e n n i d mi l r n n r c i s . i b l s l i . u e been l a r r i e d t l i e i e m - i d e ma i 11 i ph a ne s W i t h o n e e \ e ( | i t i n n . no e \ i dence lias r n i r m r d to ,-iUi'jisl t ha t . isbe. lns a cell initial es selec- 1l \ e ' \ m I he s nbser i is.i I t i ssues t h r obs Kills | a i n r l s l n n I n n s I n r mi aj i ci l t w h i c h im l l a-es mesnt 11el m m a t a T h e si lie le e \ r e | it ii ill m i n 11\ i de d 11\ t h e | il ei n a I " m i I k s s p o t s " in w inch small n u m b e rs nl asbestos fibres w u r ioimd d m m p the later stares nl t he e \ p i n men I T h e ' m i l k s s p o t s ' <oiU.nn in ms ni.u ru phiircs mid a i e perl used b \ ela In iral el \ coi l ed c a p t l l a i i e s ( L a m 1 Iibies become flapped ill t h e s e si tes a n d it a possi bl e t h a t p l e u r a l m i l k s s p o t , l oa d e d w i t h asbe s t os , inas pros i de 11ic n id ns I rni ii s\ h ich lx 111i p l e u r a I plat pi es a i id p l e m ,d mesi it hel i o m a t a all b e d p i e nt IS lies e l o p T h e t i ssue M e n t i o n s t o i n|ee1c<l a s be s t os l i l i res w e r e 11 n i e m a r k a ble. but t w o p o i n t s ma s be stressed I'hrsl t h e m t e l i s i t s ol t h e o- p a n - e a p p e a r e d t o lie I >i oa (11s re la I ed t o I he a m o u n t ol a s I ies( os pr e s e nt The a \ 11!a rs I \ ill pli n od es , l or e x a m p l e con tai ne d i.ithei l a m e a m o u n t s ol a-bi'stn- which s t i m u l a t e d the dcs eh i pn l ent o f I s pi ca I 111u m i s e i a 11 nli unat a w i t h ir ia n t t e l l - - i n 111 i r t o ( t lions; h on a s m a l l e r si al e t h a n j t h e i c a e t i s e l esi ons l o n n d at t h e m i e i t i o n sites. I n m o r e d i s t a l Is m p h n od es asbest os t i br es w e r e l e w i r i n d e s o k e d l i t t l e o r no t i ssue r e s p on s e. S e e o n d l s a s b e - l o s bod i e s w e r e l a r e l t seen It i- w 1 II r c e o t mi / . e d t h a t t h e i n c i d e n e e i >1 l l i r s r s t r n e l n i e s \ a i tes m d l l t e i ent pm ie- ol ex peri in ent a I a n i m a l s ( W a n n e r I t l l e i l t he s a m i c a d i l s p i o d n e e d in " i i i n e a pi and ill hamsters, lint t i l es l i a s e not been e n e o i i i it el ed ill l e t s ( H o l t M i l l s a n d h on m j . I '.Hi a . ( t r a s s a n d d e l i es i lie I tMi7 ) a in I I i .i s e on I s a r e a - l o n a II s b e e n - r i n in i i m r ( k ' a r d / , I !M>7 J. It is d i Hie u It t o aeei mi l t I m t h is d i II e r en ee as t In a e n n m i e, i si i in es i d e n c e t h at t he I n r m a t i o n ol a s be s t os b o d i e - is ( s s e n t i . i l ! " an 1111I ,u el 111la r p i o o s s l o l l o w m n t h e ill |("i I ii n i ( il a Ii I ii e 11sa n la ri npl nn.. e i I ) i " I 1 11M7 I I ol 1 a n d \ i i l l l i r . 11M>7 I tot ha in a nd I I oil I ' a is so i / u k i 11 el 1 h u m . I hi >' i , oh w a \ l o - n ol a s b e s t o s bs I i ssue m a i rn p| i a n o s n nis o I m m i Ins- ei teet i s e in i O ' a n d n i n e I hai l in p u n lea l " M a n d h n n - t ei s 1 h i r j ii -- i iti Il ( s w k lei i e, ili not !IM ,M i hul e ! ! 1^ 1hat asl iest os 1)0(1 II s a r e 1oi n led III 1Olt i n i ! - .Ili io pi oli! pi i - d i s t i t\ rtj l i 111lili d lie b o r n e m 111 mi l t h a t t h e s p er l 1II i! \ M| ,1 sbe !los l u i d l e - 1- 1il 11i<j nn l e l ' III li i \ ( | u e s t i n n e d (( ton nli I'll i t , , .SS dl ' r i c \ 11I r < hai ics a n d I t e s F m;,s ( . aei i sl e r a u d A d d n mt on 1nun ] < 1'lie (list u r l i n e : le 11li re 1)i t ti r p ' ( "III ex pel 1m e n 1 '\ i - t h e e 1 ein r e n e e ol w hat a p p n a red t ( , he ,.s i m`m| , i- 11iM I T III t l ' - UH' 1i on i sin ne o : i it.- lint m 1a 1ed emi t m l m i c e s mi l .a r ex pi nell' ( li.l \ ' In*'ri ! i ("i ii a t ed bs o i l i e r ms rst \il.a! o s i( H o b l i n i . 1`.Hifi ) ) O 0 K. KANAZANVA. M. S. ('. l!l U ISKl M. I. I.. CAI TKI! A \ 1> I'. .1. C. l ilil A llh o m d i eotiaidei al)l`` (.u c \\is tak e n wili all prneedures nvolvine asbestos librea 0 sriiii.^ li k e h ( l i d anmc n m I a m i n a t ion ni (he e nn tio l nnim.ils n i r u m ' i l . ( ' n i i l a n i i n a t ion l i a - a U n b e e n n b a e i x c d in \ w l d a n m a l a I n i n r iii t h e \ 11 1111\ ol asbestos imiK-i in S n u l h Af ilen ( \ \ clister. 1003) and (lie picsenl obserx al id u s . all Iuiiiu'Ii i i i \ u l \ i n " n nl\ a small proportion ni animal';, cmphasi/.c the e x t i e m e r a r c ti cees sai \ in ha n d lin a asbe s t os in t lia l a b o r a f u i x . T lm w ork reporled in lilis papor uas undcrtnkcn d iirin p the tenure (h\ k .K . ) nf` un Klcanor lioiiw a elt In ternational ('alicer Keilow ship ol (lie American ('alicer iSoeiety. awardcd liv the l :nio Internationalis ( 'nutra ( 'aiiermn. Snpport was also p r o v id c d Iiv eranta tn t he C h cstcr l ie a t t v Research I nal it lite. I n s t it u t e o l' ('a n co r Research' I!n \a i Cncer Hospital, rom tlie .Medical Research Couneil and the lr itis h K m p ire ( 'anecr ( 'a m p a i^ n f'or Research. R R I'KRRXClvS I i i KKU' A , ( ( . ' l i eun, .1.. S k m k o f i ', 1. a n o N m i t i i . \ \ ' . 17. - - ( I 00.7) . 1nn. X . A r a d . S c ., 132 IS R o t i i u i . S . K . a n d l l o j . T , 1' . R . -- ( 100S) 7 . I 'a l h . / A a O . 0 6 . 1 IR. ) v\ i s . - .M ( I 00.3) . 1mi. A' . . liv/7. iSri . 132 OS. (1007) /ir. 7. r . r /i ./ W /o , 48, 370. (! \ KNsi.Kit. 17 A. and \ i i Ii i m ;t i i n , \V. \ \ '. -- (1 !>(>!)) X>ar Eitt/I 7. Mn!., 280. 4!SS. ( i'ol . o, ( ' ( I!H)7) 7 . cl i n. I 'alli.. 2 0 . 07. C o r n i l . .1. (111(17) A u n . X . Y A r a d . Sr i . , 1 3 2 , 30X. Ci t oss. I ' , i m i o i . T i i a i i .i , i ;, R. T . 1'.-- ( 1 0 0 7 ) .1 /<7/s en r'u l i l i l . 15, 038. ( l i osa. |>., d i . T u i a i i .u :, R. T . 1` ., C k a u . k v , L. .) a m R u i s . .1. 31. < ! . - - ( RIOS) A r r h * l 'ath . 8 5 . .730 l l n i . T , I ' K , M i u . s . ,1. a m i V i i i ' m i , 1). K . -- (l!l.'>) J i m . .V. Y. A r a d . S r i . , 132, 87. l l o i . T . 1' . I1', a n o Y m .Mi, I). K . --( 1 0 0 7 ) 7 . I 'ath. I l a r t . 03. 000. I l o e in ii i m , D O ' R . - ( RI0.7) .1 mi . X. }'. A r a d . Si i . 132. 047. hvMi. !.- (1002) V.I Ufo, ,rh. i i i iYm.ik. A n a l . . 5 8 . -1S7. M i o t t i , R. ( I Ola i .1 <ia a na l . . 6 2 ts0. X i i M i m i sic. M, h. \ s n T h o m c s o n , I I . -- (100.7) . I un. X . Y . A r a d . Sr i . , 1 3 2 , 5 7 0 . I ' k u 'o c k , I ', l - (RIOS) L a i u i l . i. 1153. I ' k a c o c k , I ' l. a n o l ' K i m i k . A - ( I00f>) . | mi. X )'. . \niil. S r i , 132, all I ' h k s s m \ \ . .) .1. X M i S l M o s . M 1!.- ( 1 0 0 1 ) S m i j r / ' i / l l i / n r r O liArl.. 1 1 3 . 7) 37. R o n , I-' ). C . ( A i i t k u . R. L., W . u / m t s . M A a x d 11 u i n o t o n ', J . S - ( 10 0 7 ) / n i . 7 . < 'a u n r, 2. 02N. S k i .i k o i r. I. .1., O i i n u i , .1, a n d H n n m \ i i . 17. ('. (100.7) A' n r Kntjl. 7 . M r d . 2 7 2 , 7)00. S n i r i i . W 17.. M u , i .i ,i; h . . ( ' m i i n 3. w n Si n i k m r, I, .1. ( 100 . 7 ) 7. M l S i n a i I l osi . , 3 2 , l . S u m . \ V. 17, M u . i .i .ii k . I7i.s vssi.u, R 17 a n d I I i u r i t T , I), I ) . -- (100.7) A u n . X ) ' . .1(7/7 Sri 132. f7)0 S r n i i ' i i n s. J , \ x n M i . \ i n 1' l . -- ( RIOS) .1 un n m i p lli/i/., 11. 2S3. N i / , i k i . Y i m i ( ' m e o .1 ( 10 0 0 ) , 1ai 7 l ' a l l t . . 5 r i 70. U ' u . m . i : .1 ( ' ( I 0 02) A a l i i i i . I . aud 106 I m i - -(1003 l r. 7 . i nd. M r d . 2 0 , 1.-- (100.3) . l e / / A , 1 | ( i / 7 Sri . 132 7)7.7. I V u : \ i . i : . -I. O , S i . r c c s . O A.. w n M \ i : m \ x n. 1* ( I 0 0 0 ) / k 7 i nd M r d . , 2 1 , 200. U ' kk ' n. U. I ( 10 0 3 ) A a t u n . L a m . 107. .700