Document 3e7J9JXgZq7EJ1vwydwode8BE

IF] EXHIBIT 373 Chromosome Studies in Workers Esposed t o BenzeDe or Toluene or Both I/wandra Forni. JID;Emma Pcsifico.ScD; ond Antonio Llrnonta. M D .Jfilnn. Ilaly ;nosome studies were carried out on .-ral blood lymphocytes of 34 workers of a :.ure plant and in 34 controls matched I md age. Ten of the workers were exposed ..:me before 1953 and then to toluene. and been exposed only to toluene after 1953. -:.;portions of unstable and stable chromo- some aberrations were significantly hisher sta:istically in the benzene group compared with the controts, and in Ihe benzene group in comparison with the toluene group No significant differences were tound between thr frequencies of chromosome changes in the toluene ud control groups. .;(]SIinChalation of benzene vapors'is uused by the toxic agent, provided the expo- . :o be able to induce toxic eirecrs on suze to other chemical or physical agents .:arrow, resulting in pancytopenias, capable of inducing chromosome changes is ..ineventiidly evolve into leuke&.1.= d e d out. rytotoxic e5& oi benzene however III 1952 to 19% an epidemic of benzene- :.I( seem to be restricted to bone poisoning occurred in a rotogravure plant. .L cclIs, but also involves other cell where benzene was largely wed as an ink &era1 recent reports have appeared solvent and diluent. Many workers suftered ,.h chromosome aberrations were in- fmm severe benzene hemopathy, many i ti! occxpatiod e-rposure it0 benzene showed slight heaatologic bnges. and only wt only in tone marrow celis.3-5 but a few showed no sign of to~iary.9Since then 1 [tic lymphocytes of peripheral blood. toluene (containing traces of XyIene) was rcsed rate of chromosome changes in substituted completely for benzene. :..:llpilweS cultured in vitro was dem- The-present investigation, part of a wider not'ody in subjects with benzene study on chromosomal damage from benzene. dhy:.' but in subjects recovered from was carried out to study the frequencies of hemopathy,d and in subjects ex- c h m ~ x n eaberrations among workers of :.:,I.1 ...aI-mme withut overt signs of blood the Same rotogravure plant, some of whom had had known exposure to benzene beiore ilimmoSOme aberrations observed are 1953 and to toluene after that time, but most -I.ific, similar to those induced by x- of whom have been exposed only to toluene. . i d a n persist even several years The mtes were mmpared with those o b . I -ition of exposure. They may, there- served in a p u p oi matched controls. -n p r c x n t a sign of cellular damage i t 4 for publicanon March 31. 1970 ac. Materialsand rMethd iiml 16. ilr clinica dol Lavoro r U i g i Dwota" I\ of hlilaa, S t i h . Iuly. :it rqiesm to Ciiica del Lavom "Luigi Viu S Burubr 8. a122 Mi& ItAy (Dr. The subjects studied in this surwy were 39 male workem, who had been occupationally exposed to benzene, or ita homologues (toluene with traces of xylene), or both in a rotogravure Arch Enuiron Heald-Vol 22. March 1971 .- 374 CHROJ1OSOJ:E STUDIES- -FORXI .ET AL Table 1.- - Wori \ 3a OS 45 54 41 36 05 50 * ?4umoersin oarcntt t a b k Z-EXC Y Exc -IC Age 1 29 - -30 32 41 32 u_* .. .. 1 -.. - . *J - t -- I - -- I .. 42 35 39 47 30 4a 38 33 34 60 45 43 55 58 TaDie l.--Exposure Data. Chromosome Studies, and Hcmacological Notes Of Tan Worners Exzosrd to Ben:ene detore 1953 and to Toluene After 7953 ..,+ Years 01 Exoosrire to Total . Ceilr &je Benzene Toluene Sturidd 38 <1 1 4 I00 --% C.llr C. c s 1 ... ...15 2 1.00 2 2 ...45 3 14 64 .*. ...54 3 14 100 .I. 54 3 14 t00 ... ... ......i l 3 I00 2 47 >3 1 4 I00 3 36 5 14 100 45)' 45 7 14 100 2 50 22 12 100 45)' cu'.mocrs in parentheses ina.i-ca!a the calculated brranr in cells. 1 ... 1 ... Hematolo@ical%acts Shgnt anemia in 1953 ana 1557 Severe ocnztne nemaoatny in i 3 5 3 Sii@htanemia in 1 9 5 3 . 1 w . i g s a . :96a Slignt anernla ana ieuuaoen~ain i953 Sbgnt anemia in 1955 .Severe benzene ncmoparny In 1953 Slight anamca in 1953 Siignt anemia in 1954 ... ... Table &--Exposure Dam, C.?romosome Studies. and Hematologicd Notes of 24 Workars Exposed Oniy to foluena Lutsr 7953 - Aar 29 20 32 41 32 35 32 31 * 35 37 42 35 39 47 30 48 38 - 33 34 -. 60 .. 45 43 - 55 58 Years of Exoosure KO tolucm 3 7 7 7 7 7 7 7 8 :0 10 10 10 10 1: 11 11 ?1 I1 12 13 14 If 15 Total Cells Studied 100 100 100 I00 100 100 too 100 100 100 100 100 100 100 I00 100 100 100 100 I00 100 100 100 100 % c4u. C, c; .1.. ... ......... ... ... ... 1 ............ 2 ... 3 ... ... ... 1 1 ............ .2.. .I.. ... ... ... ... 1 ... 2(3P 1.. 3 ... ... ... 1 ... ... ... 11 * '.mumr tn parantneser inaiuter the calculated Dfeaus in Cu a h Wemato1og)cal Notes . ...M ~ e r a I etnromoocytooenia tn 1565 ...Slignt anemu in i96i. 1963.:566. 6967 ... ......... ... ............ ..* ...... ... Slight anemia in 1961. 1963 Siignt anemia in 1957. L460. :962 ... ...Sbgnt anemia in 1558 Slignt andmi. in 1955. 1966 Shgnt anemia since :956 Mooerare I e u ~ o ~ e n tina 1960 Arch Enuiron Health-Vol 22, .kfarth I971 6.3 :?5? . - -:95a . C~ C J :?i3 223 56 ... 189 234 200 2:s 226 -. 1361 1L O -. i 962 239 186 277 1963 150 213 1964 200 239 :965 !957 ...2:3 239 156 Mean values o fmany dsrerminauont. 513 306 322 e29 43 : a24 338 345 372 265 in their nzztked mntroLs are repoced in Tab:es 1, 2, and 4. A11 &ta ye summarized in Table 5. D3ta were d y s e d statistically by the x' !est and, when possib4 by the Wilmxon rmtdied-pairs test. Benzene Group Versus Controls-The incidexz of unstable c'xomosome changes 11 12 !3 25 28 32 1 100 ... 2 100 ... 3 100 ... tC3) (1.665)and of ulcrrlated breaks in 14 41 4 100 2 C,,ceils !l.S%) were higher in the benzene 15 32 5 100 ... ,-up compxed with controls (0.61 and 0.675, respectively); the difference \vas highly si,w5unt (2= 8.45, P < 0.01 for C3cells; 12 = 9.96, P < 0.01 for calculated breaks). The same was true for the kgren- (e-cy of stable chromosome changes 7.51, P < 0.01). If one exdudes from the benzene group t!e two subjects (cases 2 and 6) who had 16 17 18 19 20 21 22 35 33 29 35 37 4.4 35 6 7 8 9 IO 11 12 1GO ... .1cc) : :CO -r&.9-. ,. 100 ... 100 1 100 1 100 1 s i i e r e d h r n sewre benzene hemoixthy in 23 36 13 100 l(3: 1953 /.see Tabie 11, the frequency of C, 24 46 14 100 I ...ceh..w3s l S % ; that of calculated breaks in 25 29 15 100 C, cells, 2.09%; and that of C, cells, 0.26%. - -Such ~roportionsare significantly higher than those for controls for C, oeb and d- cula+d breaks (9 9.53. P < 0.01 for C, cells: x' 11.87, P < 0.01 for calculated breaks in C, cells). The rate of stable c h - 26 27 28 29 30 46 38 32 34 60 16 ,100 17 100 18 10C 19 35 20 100 2 ... ... ... ... -macome changes, though higher, did not difler significantly from that of the control group (,? 0.39,P > 0.05). Toluene Gmup Versus ControLS.-In the 31 32 33 41 42 51 21 22 23 100 2 100 1 100 ... toluene group the proportion of C, cells 34 55 24 100 ~~ ... (0.3%) and of ulcuiated breaks (033%) was sornewnat higher than in the control group (0.61% and 0.67'5,respectively), but the differences were not statistiuily signif- Cases 1-10in "Watcnca Wlth" coiumn y m a as most in Table 1. t Case 1-24 In "Matched With" column I . . s-~a m e as t h o s e in Table 2- t Numbers in Parcntiesas indicate the ca breaks in Cu CCIIS. Arch Enuiron Health-Vol 22. March 1971 Table 5.-Chrc - 2 for three pairs.). I*.iliicb 6 presents 3 s m ' tlw chromosoze coun :IIW The proportion oi Ar .4L - Total ::-ea Cells vltn Stuaied --- ":. - I . . C, ..e+ :CIS e x p-o.s e d lo oenrciit- h ! 1 :co ... 2 I00 ... 3 I00 ... 4 62 ... 5 65 ... 0 100 1 7 100 ... 8 100 2 9 ICJ 1 10 100 2 iects exposed onry 10 t o l u v ~ l r ' . ...:to 2 100 ... 3 xco ... 4 :GO 2 S 100 ... 6 100 ... 7 :23 1 8 1GO u2): 9 100 ... ....10 100 Ico 1 A 12 I GO 1 13 100 l(21: 14 100 1 ...15 100 16 100 2 17 100 ... ...18 100 19 35 ... 20 100 .I 2: 100 2 22 I00 1 23 100 ... ...24 100 3 "Haccnoo With'. :dole 1. i "Hatched With'. :able 2. coiurnn .I?column .I#- ' i 1972 Table 5 . 4 r e Q u e n t v ot UnstaDte and ; : d e C3romorome Changes in Sub,ec:i E x p x e a to Benzene or Toluene or Som. d n a in 34 Canfrors Eroorure Sublects No. of Cases Age Ran3e Total Ceris - % Cells 0 - .' c u C. Srnrcne (f toluene) Toluene Cantroi suapxts 10 24 34 36-54 29.a3 i5.m 56A 2.403 3.262 1.6txi.a7~ 0.80(0.83)' O.Si(0.67). Numaers in parencneses snow percent of caicuiaced breams in C, c e i k c 62 c.ca 9.;4 fable 6.-Cbromotome Cauntt in Su6jec:s Exeosed to Benzene or Toluene, or Both. and in 34 Csntrols Exposure Sublccts ~ No. of Cases Total Calrs Csunted % Calli With Chromosome Nomoer <46 46 >46(PO8yp1OiC) (+Berueno toluone) 10 964 13.1 86.0 G.g(O.52) Tduono 24 2.400 14.3 85.4 0.3(0.29) Control suniects 34 3.262 10.2 89.5 O.j(O.3) -: cxz 0.43, P > 0.05 for C,, c e k was similar in the three groups. X siigixly I K ? ~ ,P > 0.05 for alculated breaks). higiner rate of cells with more Clan 46 cbro- nte of C, cells was very similar in the ~ O S O w~as~ oSbserved in the knzene group. x group r0.08%) and in the controls This fad seemed m d y due to a higher mte *I";), and the differencew8s not sgnii- of poiyploid cells, but the dXerence was Got >: 4 12 = 0.11,P 0.05). !:.rumGroup Versus Toluene Gmup.- -statistidy signiiiont (p=O.a, P > 0.03 frequencies of chromosome changes in - -! + m e group were higher than in th? Comment ..w group, and the diierenceswere sta- .9-nilysigmimnt (x' 4.22, P < 0.05 for The increased proponion oi ciuomsorne w~L;; f 5.68. P < 0.05 for d a b t e d changes found in the subjects e m s e d to Ls; = 6.30. P < 0.05 for C. cells). benzene in the past is in agreemzt xith C-2 data obtained in the two mups of other reporis.4.' Cornpared to *hdata oi *-w d subjects and in their rritched con- Tough and Court Brown,' cur series showed '- (* Tabla 1, 2!, and 4) W e e a lower incidence of stable &mmosorne l i d by the Wilcoxon matched-pain changes, both in those exposed and in *&e - The frequent o c c u ~ ~ ~ofc peairs with controls. This may have been due to dii- . srme score greatly redurd the number ferences in methods used to asses stabie ' - . t i 6 useful for the analysis. However, the aberrations; in our study much of 'hwork --nrtions of unstable and stable chromo- was done d e r direct obsemahn, so t.ht 2 changes in the benzene p u p were minor chromosomal ohnges. espe3ally in .-:fimtly higher than those in ~e con- at the signiiicance level of 0.0% ic,, -'.. t 2 for seven pairs; C, oells: t = 0 - !our pairs). &rornosorne m"roops B. C, and E might not have been detected. The group of subjecs exposed to to1uer.c did show a somewhat higher rate oi -able iI.,icrenees in the proportions of C., and chromosome dunces snd oi c a I c u I a d d l s becween the subjects e m d to breaks compared to the controls. but the MC and the controls were not s i a c 3 n t differencewas not statistiuIly si,nni;iunt. To c&: t = 36.5 for 13 pki; c, d ~ o:ur knowledge, no data on the d~romosome for three pairs). status of workers exposed to toluene are ;'hlc6 presents a summary of the results available in the literature. s :hc chromasome counts in the three In rhe groups oi subjects expos.& to h- The proportion of aneuploid cells =ne, or toluene. or tnth the values were Arch Environ Health-Vol 22, .Ifarch 1971 ___ - -_ 37s I CHRO.'.iOSO.'.:E STYDIES-FORSI ET .4L Re!erences 1. Cron;d!e EP: Evidence [or mbation ana chcm- icah IS kukcmoqenic agents. Arch Encimn Hrocth 3:297203. :?5L 2 V i g l i n i EC. slih C : Benzene and laub.c mi& xexl Yorh J *Med 271m576. LW. 3. PolGni G. Colombi R: I1 d o ~ m o s o m i c o midollare nciI'ancmia aplasuea bentolin .Wed Lcl- unm S.i::'41-253. 1964. 4. I ' o m i A: Chromollorne chances due to chronic exposure to benzene. in Prwrrdurer ai rhe i5th Intcmaiiono! Conqrrrr on Oceumtwnd HaitA Vienna. 0c:ober 1966. Vienna. Yicner Mlcdjzi~~ircire AJmdernie. 1%. vol2. part 1. pp 4.33749. 5. Fomi A. Morn, L: Cftoeentcic rw&a 51 a a= of benzene Ieukacmis. EUMPJ Cancrr 3251- 253,1357. 6. ?o!:ki G. Colombi R ?! danno c r o . ~ c x i c o dei linfmiti nell'cmopatia h e n i c a . Mea Lorvm 55:M 1-63.!W. -.% ' J- L .. ..,. ._ 1 -,. ., ~ . ..~ . . I-- .'*? cllectr of nitrogen c system of the rat J studies were mad ' )I? directly into the ".c. The rates of flaw o : 3. 3.0, 2.5, and 2 0 lite d: that there was incrE -J ' 3 exposures of the !Is: A! of NC, introduced : was lowered, there wz - w e e between expos - -7s. Exposure at flows 9 had an insignificant ' #Icy. \ I.IIOiASE is an int 1 !i is found usually in cyoplasm.* In n o m :6 ) 3 smail amount'o ....:171in the serum ot : I!tioIoc+ai condition-. is elevated.24 When ! !i?rfact LSat certain .hly lead 'to patholog. -t.nsibly suspect alte ..it? due to air pollutar i i nn air pollutant u d ! r nnoke and in urt - p l i ~ r ~asnd which h: zttlon for several ye: Iiurn" considered the 6 rrlmon tissue folIowing -:wwure of live guinea F Jnwtigations. there wa 1.1-v activity due to .el. was also increased . which could be due m z the release of the ti: -. ' . l , l l W i . y. . ~ t ~ t i t e tllor publiration S i n :I. '1% ilic Department of P h IN.I**nirlciahDiekinJon L altrt rqwsts to D c p r t m -..;,it Dcntiotrv. Fairleiqh Di 2 , .UJ 07666 (Dr. Ramam Arc