Document Nzn6Nd2Ky40ZaD4nYKV5YnRy

R&S 109769 BIO-MEDICAL RESEARCH DOCUMENT DESCRIPTION FORM 63 68 69 76 Duplicate in all cards: --> i^S-o 03 0,3/25 6 year as-1961- File number [Right justify [Numeric only] Author(s), as Last Name FS (No Punctuation) and coden for journal as JAMA preceeded by one blank space fec/tt'L' 20 21 H') .5 fi-l 40 41 77 78 Sub-Index Code 60 61 62 11 12 13 Title of Report; end with space-hyphen-hyphen-space. Follow with Index Terms, separated from each other with comma-space. Avoid other punctuation; do not abbreviate. 61 62 21 22 23 24 Source (Journal, Vol., Number, Pages, Date) 12 (0 h t.Q i c ct m1 cm Ih ft r ma-'H / r n<? n fn lozhcni^ Alp. 3 . pp P</- 5?, if^LilU -hh , )cno 61 62 31 32 Brief Summary 12 10 SUMMARY: 61 62 61 62 63 64 256 #11 INVESTIGATION OF THE BLASTOMOGENIC ACTION OF POLYVINYL CHLORIDE RESIN USED IN THE FOOD INDUSTRY. M. Radeva and S. Dinoeva Translation of: "Prouchvane Blastomogennoto deystviye na Polivinikhloridpa plastmasa, izpoluzuvana v Khranitednata Promishlenost," Onkologiia Informatisionen Biuletin, Prilozhenie, No. 3, 1970, pp. 24-28. R&S 109770 ifXI. INVESTIGATION' OF THE BLASTOMOGENIC ACTION OF POLYVINYL CHLORIDE RESIN USED IN THE FOOD INDUSTRY M. Radcva and S. Dinoeva Translation of: "Prouchvane Blastomogennoto deystviye na PoXivinikhloridpa plastmasa, izpolzuvana v Khranitednata PromishlenostOnkologiia Informatisioncn Biulctin, Prilozhenie, No. 3, 1970, pp. 24-28 A number of papers have been published in recent years on the development of tumors in animals on subcutaneous implantation of polymer plastics. The occasion for research in this direction was provided by the malignant tumors obtained by P. Turner in subcutaneous introduction of bakelite discs. .The blastomogenic activity of synthetic polymers was subsequently confirmed in experiments by Oppenheimer, Druckrey, Hueper, Benerjee, Bering, Laskin, Nothdurft, N., Kogan, Shabad, and others. The essential nature of the blastomogenic action of plastics has not been clarified. Among the various hypotheses advanced regarding the mechanism of such action we shall deal with the hypothesis of the chemical mechanism of the blastomogenic action of polymers, since it provided us with the occasion for the present study. According to this hypothesis a blastomogenic effect is exerted by the active radicals separated from the implanted plastic or by the active chemical centers formed after the separation. It is assumed that these radicals, in penetrating the cells, cause.depolymerization of the nucleic acids and modify the activity of certain enzymes. A blastomogenic effect is also exerted by the depolymerization products separated from the macromolecular component of the plastic. The hypothesis of a chemical mechanism of "polymer" blastomogenesis is favored by the experiments of Oppenheimer with subcutaneous implantation of labeled 14C polystyrene-, polyethylene, and polymethylmethacrylate, in which radioactive substances indicating depolymerization of the implanted material involving separation of low molecular fractions appeared in the urine of animals during the 21st, 26th, and S4th week after introduction of these materials. It is known that depolymerization processes may occur in plastics in the course of time as the result of "aging" and when they are used in the food industry. In this connection the question arose of the possibility of blasto mogenic activity by dcpolyraerization products when introduced into the organism per os. Study of this possibility, along with study of plastic and polymer articles intended for the food industry from the toxicologic standpoint, is necessary for the sake of more thorough characterization of such plastics. The purpose of the present project was stqdy of tlie action of polyvinyl chloride plastic on the organism of white rats after subcutaneous implantation and administration per os in the form of an aqueous extract. Materials and Procedure The effect of the polyvinyl chloride pia Stic employed in the experiment was studied with lie white r.n:. of no specifi c strain havir g an initial v.c.e.ht of 120-130 g and divided into 5 groups. -1- R&S 109771 One group of 30 animals was treated daily through the drinking vessel, with an aqueous extract in the amount of 10 ml per rat. A second group of 30 animals, which served as the control, was given only water. The treatment of the animals began with an extract prepared in advance with a plastic-water ratio of 10 cm2/ 1 cm3, and the-contact was continued for a period of 8 months. This ratio was maintained until the end of the experiment, by the daily addition of water in an amount equaling the amount of extract consumed in treatment of the animals. Extraction of the plastic was carried out under [Translator's Note: illegible word] conditions to accelerate the "aging". The weight, the blood picture, and the catalase activity of the blood according to the Bach-Zubkova method were investigated in the course of the experiments as indices of the action of the substances extracted from the plastic on the organism of the animal. The blastomogenic action of polyvinyl,chloride plastics after subcutaneous implantation was studied with a group of 30 rats. The polyvinyl chloride discs, having dimensions of 1.5/2 and a thickness of 1 mm, were introduced subcutaneously on the right side of the back, under sterile conditions. The control was represented by 30 rats in which glass discs had been implanted in the same manner and another group of 20 in which no discs were implanted. The animals were observed for a period of 10 months. All the animals which died during the experiment and those killed after the experiment had been completed were analyzed from the pathomorphological standpoint. Results and Discussion Our experiments confirm the data in the literature regarding the blasto mogenic activity of polyvinyl chloride when introduced subcutaneously. The first tumor appeared during the 11th month after implantation, the number of tumors increasing to 5 by the 14th month (see table). The percentage of blastomogenesis as calculated on the basis of the number of animals remaining alive after the sixth month following implantation (which was adopted as the minimum latency period of tumor formation) is 21.Vi. The tumors ranged in size from that of a walnut to that of a goose egg, had rough, surfaces, adhered to the skin and the underlying tissues, and were of solid consistency. In the case of two rats the skin covering the tumors was broken (2 tumors of a size of 0.5-1 cm, bleeding, with uneven edges). Incision of the tumors revealed the cavities in which the polyvinyl chloride discs had been situated. Microscopic examination of the histological preparation revealed 3 fibroblastic sarcomata, one polymorphous sarcoma, sind one adenocarcinoma. No tumors were observed in the control animals. The group of animals treated with an aqueous extract of polyvinyl chloride resin gained less weight from the 3rd month to the end of the experiment than did the control group (see Figure 1). Investigation of the catalase activity of the blood revealed lower values in the experimental group than in the control group, the lowest values being reached in the third and 13th month after the beginning of treatment (see Figure 2). This inhibition of catalase indicates suppression of the oxidizing processes in the organism in consequence of introduction into it of substances extracted from the resin. No tumors or changes in the internal organs were observed in pnthoinorphologic.il examination of the animals treated with the extract rer os. R&S 109772 C/.2.60V ** 1 * / 1 TClT no r^ncTiiuKtt CTiKACUa TABl.F. 23 itq -T DG CV'.'i i:*4- to 10 20 IS 4 ,10.1 ma 5 >f'.os;C7- l ;i:;j ;iocr Tj'JOpil 5 27,7 rp. 120 20 - - Key: 1, Type of implantation; 2, number of animals in experiment; 3, animals alive 6 months after implantation; 4, number of animals with malignant tumors; 5, percentage of blastomogenesis; 6, PVC discs; 7, blast discs; 8, control group. 33 9 03 Findings Polyvinyl chloride resin introduced subcutaneously into white rats in the form of discs having the dimensions of 1.5/2.0 resulted in the formation of malignant tumors in 27.7% of the experimental animals. An aqueous extract of the same resin introduced per os over a period of IS months had no blasto mogenic effect, but the toxic action on the experimental animals is to be emphasized; it was expressed in suppression of the activity of the blood catalase and reduction in weight gain. / V R&S 109774 BIO-MEDICAL RESEARCH DOCUMENT DESCRIPTION FORM 68 69 76 Duplicate in all cards:--> ! cl!fO year as-1961- File number [Right justify [Numeric only] Author(s), as Last Name FS (No Punctuation) and coden for journal as JAMA preceeded by one blank space 1 20 21 /CC/chlm rVT 1 40 41 77 78 VC Sub-Index Code 60 61 62 11 12 13 Title of Report; end with space-hyphen-hyphen-space. Follow with Index Terms, separated from each other with comma-space. Avoid other punctuation; do not abbreviate. 12 r _rH; lV_ i u 1p irV n-f- (< id } !; r,) n i~ 61 62 21 Tra e Cv h ne e f7 ilc T i 5 ^ < c 22 (vipScdt h Arc ((,*\cf i 'v>l p fn /rrf-F-~ 23 ____frf-- Pi a 4 tic___ r i a 1------ 24 Source (Journal, Vol., Number, Pages,Date ) 12 \/epr- cntd. l/-(. "7. /J.-' c(. op. is-/i, ---------- tllsl----------------------------------------------------------- !---------------------------------------- 61 62 31 32 Brief Summary 12 10 SUMMARY: 61 62 61 62 63 64 R&S 109775 Vopr. onkol. 7 No 9, 13-17, 1961 Or THE DEVELOPMENT AND MALIGNANT TRANSFORMATION OF CONNECTIVE TISSUE CAPSULES FORMING AROUND IMPLANTS OF PLASTIC MATERIAL* N. T. Ratkhun and A. Kh. Kogan Lk-.-aidry of Pcrhomorpholocy of the Institute of Experimental AnJ Clinical Oncology of the Academy of Medical Sciences, and Department of Pathological Physioloy.y of the Firjt Medical Institute, Moscow (/Zece/Yed 27 1962) ** a<pect of great interest In recent years in the sphere of cancer research "u k-'en the study of the blastomogenic action of a number of plastic *CfJJ`i (cellophane, polyethylene, polyvinyl, chloride, teflon, polymethvlII|*t1Cry!a;tf, polvstvrene, nvlon, pcrlon, caprone, dacron, silastic, jarane, V^0-', pliofilm etc.). Mute the main attention of researchers [1-5. 7-H] has been focussed on w) of the properties of plastic materials (chemical structure, physical torm) Possible for the causation of tumours, the features and patterns of the Piufes ot cbnucctise tissue developing around the implanted plastics and the onLoL 7* No. t, 13-17, I9*K > 12S8 N. T. JtAir.miN jml A. Ki(. Ktir.AH processes of their malignant transformation hov/cvcr have remained a!niolt entirely without inv>-.ii;;.nion. M.tU ri.tl jml trt /.i/'/hi*. I.xr,-rir,,cm- were performed on white itockbrcd frmite fJ wri;;li;i>r. IC3-123 K- A tomprc'.'ivc capsule of polyvinyl chloride waj affixed aro-..,j tj|* kiilnvj'. Tin- rus were .iinfi*.vd it 3, I 3, 15, 33, '*3, IV5, 28j, 330 and 383 dtyt, imp!.int.u|.in. with 7-13 r.*;s in each of the ^roup*. A total of 83 animals were uicd. j:M.^ wive fiscd in CarnovS fluid and inoontcrl in paraffin. Sections were stained with haentat;,!t-': rosin and picrofuch.in aceorjini; to Van Girson, and impre:;natcd by Gomori't Poljiaccharidet (p.lyeo.v/n, "lycoprotiins and muenprotcins) were demonstrated by s.h:,v, reapent after treatment of the sections with periodic acii with.control treatment 0f secs'- with amylase, "and acid polyaecharidcs (mnioprotcids) were stained with toluidine b!jj aleian blue, f or their further differentiation before staining sections were treated wish t vie-' * and bacterial (streptococcal) hyaluronidase, auJ alto with toiuidinc blue at various pH VJj Collapen.nc was used for identification of scleroprotcin fprocollajen). ll In addition, Tceljen's reaction seas performed for desoxyribonucleic acid (DX.Vi. ice ribonueleie acid (RNA) was 'stained by Crash's method with pyronine-methyl green, yy..'' was accompanied by control treatment of sections with desoxyribonucicase and ribosadtiv Results of the investigation. The jniri.il reaction around the implant it w aseptic inflammation. Phenomena of proliferation- rapidly start to prcdomi.-si*. the young fibroblasts being rich in ribonucleic acid, by the third day op.* Ci. already observe in the capsule forming around the implant the appciriaee of fine argvrophillc Schick-positive fibres. The reaction of merachroma*-! it extremely weak during this period. At the 10-15th day there is a capsule rich in fibroblasts and consistin' of collagen fibres which stain intensely with picrofuchsin, arc weak Schkl. positive and do not give mctachromasia at pH = 2*0, pH = d-C j-: pH -- 6*0. At this stage the connective tissue capsule has-a non-homogeaeoj structure in various areas. In some places it consists entirely of collagen aligned along the implant (Fig. 1), the cell units being fairly regularly disposed and consisting of both young and mature fibroblasts. In other places ti: selfsame capsule has a more complex structure and consists of no less the; three layers: external, middle and internal (Fig. 2). The external layer is n.: in connective tissue cells. The middle layer consists of both thin and :h:;i collagen fibres running in various directions, sometimes having the appjjrrrw of dissociated bundles. These fibres stain irregularly with picrofuchsin, i are sometimes markedly and sometimes feebly Schick-positive, without star of mctachromasia at different pH values. Sometimes one finds amor; u; fibres in the middle layer focal accumulations of Schick-positive no.i-nu'j chromatic matcriai. In the middle layer of cells, the vessels which as a rai are not numerous arc arranged irregularly. The inner layer consists of eell ;;: 1 fibres aligned along the implant, feebly Schick-positive, and staining red <-- : picrofuchsin; it contains mature fibroblasts and flbroeytes located length*-* along the fibres; there arc almost no vessels in this layer. The thicknessei layer can be exceedingly variable, but the middle layer is generally ths-n-"- .- By the 3Cih day the capsule consists of fairly mature tissue. f R&S 109776 Devclupmcnt and malignant transformation of conmctive tissue capsul'.-s 12-tV /f:tr three months tlic collagen fibres in some parts of the capsule acquire ir.aticJly ir.cta-ltromatic reaction, more pronounced at pH = 2 and staining ptalili with alcian blue. The metachromasia is abolished by tcsticuiar and ettr.aited by streptococcal hyaluronidasc. In parts of the capsule with ir.-tpiar structure some of the fibres gi\*c a positive reaction of rnctachromasia, neliihed by testicular hyaluronidasc and unevenly attenuated by streptococcal ** connective tissue capsule consists of `^tolhgca fibres. Van Giesoo. Magn. X DC. Fig. 2. Capsule of complex structure, consisting of three layers: internal, middle and external. Haematoxylin-eosin. Magn. X DO. ^utonidase,-while some arc not mcrachromatic. In addition to the fibres, "f ,ctlchromasia sometimes occurs in the form of irregular patches in l^C rc^at`oa to hyaluronidasc tn these sites is highly variable, the ':hromaiia being sometimes abolished and sometimes attenuated. The ^res and the amorphous areas likewise stain irregularly with Schitt's *1' ' ia^ somc them arc Schick-negative. One discerns a variable rela4 s^c collagen fibres in areas with ordinary and with complex structure " caiyme collagenase. Collagenase markedlv attenuates the staining with ` :j* * lU!a 0t C0'-3Scn fibres in areas with complex structure, especially in aj** haver, and hardly affects staining with picrofuchsin of collagen tfcin *tcas with o---rdin.a.rjy structure.' *~2eJCr r^rce months one can observe in the capsule, in addition to hya- -.j 0{lrai f'oor >a cell elements, scattered areas of accumulation of young n,3turc fibroblasts, as well as small cells with round hypercltromlc t*j snd mast cells. The fibroblasts contain a moderate amount of UNA, ij a'ct!me their cytoplasm contains Schick-positive granules, abolished Jsc. Metachroinasia is extremelv variable in the areas of proliferation R&S 109777 R&S 109778 1250 N. T. I'.Air.im-/ and A. Ku. Kut.A.s'-. at this stage. J'r'ilifcration of ;}i; connective ti.v.i-.- ceils or :ur n <n a;:. of the capsule with irregular structure and especially in the 'vcr. : By 6i/-i month*, the cap uic retains a non-homogenec Afrl with ordinary st'-ecturc are markedly hya!ini/eJ, and as a ml? jr in i; ly well ditferreac. tho.e ilci.'ribtj three months. At the sixth month one can obscr n the capsule ti.. 'development of focal proliferates (Fig. 3), consisting of po!> morphous nective tissue celts rich in RNA, and a few mast and lymphoid cells. Or.? it: discern in the structure of theta areas a marked m-eiachromasia, r pronounced at pH = 2 and completely abolished by testicular and attenua:;t by streptococcal hyaluronidase. Alcian blue stains these sites a green color which likewise is abolished by testicular and is attenuated by streptococci hyaluronidase. Collagenase in"somc places completely abolishes the s:a:r.iti with picrofuchsin and in other places attenuates it. The Schick-reaction in it* stroma of the focal proliferates is unequally pronounced. On impregnat.*'1* one finds an argyrophylic framework between the cells. Some focal prolife.-:'.;- are clearly demarcated from the surrounding tissue, while others hive it indefinite boundary. At the 9-12th months some capsules show over their entire extent a pronounced hyalinosis and occasional fibrocytes in the field of view. In so"-* ' Fig. 3. Formation of a focal proliferate in t!ic selcroted C.ipjulo. I lai-in-itoxylin-rotin. Maiu X no. l:tc. S, |`iltCknaionia CkMOotins out attpical cull.. I laciuatox) oi'-o'-,n- - X5CJ. ! Jjcvcfopfncftt and m-ili^nant transformation of confU'%(ivi* ti*;oe C4j`.u*cs 1251 R&S 109779 ;;'jlcs however one still detect*, signs of proliferation of connective :i*-,ue !s artti the formation of focal proliferates. In six out of 16 animal, which .rvived for 9*/i-121/! months sarcomas were discovered (l it;. 4). ' Discussion. As is seen from our data, in the period preceding the appearance f the tumour in the connective tissue capsule, one observes profound hinges with proliferation of cell elements. For example, b>* the third month bre is a marked change in the polysaccharide metabolism of the connective bee capsule, and one observes a marked increase in mccachromasia. The bding of most pronounced mctachromasia at pH13 2 and the least at pH: -6, srr.plete abolition of the reaction by testicular hyaluronidasc and its attenua:oa by streptococcal hyaluronidasc, and the results of staining with nictan dee enable one to state that in this period there occurs a rather disorderly ^cumulation of acid polysaccharides (mucoproteids) both sulphuretted and -*ose containing carboxyl groups. The irregularity of accumulation of ac:d aucopolysaccharides continues right into the late stages. The reaction of "ttachromasia is especially intensive in focal (presarcomatous) proliferates, .lathe tumours the mucoproteids accumulate mainly a: the periphery. It is fOw.ble that the abundant accumulation of mucoproteids is due to their "ptrfcct elaboration and utilization. The irregular and temporally unusual ^cumulation of mucoproteids points to a deranged collagen formation in the connective tissue capsules forming around' the implant of plastic material. ! i Also pointing to an abnormal collagen formation is the action of coilagen*'* *'Hich abolishes in some areas of the capsule the staining with picrofuchsin '?ific for precollagen, which suggests that these collagen fibres are of chnormal structure and different from normal fibres. i! . Stress must be laid on the non-hcmogcncous structure of the connective - f `;'VJC capsules forming following implantation of the plastic material: more f r,mp!e. reminiscent of the structure of the capsules around foreign bodies f 'n ^roe areas and complex irregular structure in others. Thcie atypical areas characterized by a complex structure uneven and unusual in time and ^ount, by accumulation of sulphuretted and non-sulphure:ted acid poly'-'char:Jcs (mucoproteids), by the characteristic relation of the fully formed '^gen fibres to the enzyme eollagcnase, and by the development of unaligncd v0,bgen fibres and of extensive fields of hyalinosis. Analysis of the different ^gts of the development of the capsule shows that the focal proliferates ('fIop most intensively in those very areas ol the capsule which have an J^usua] and irregular structure. This fact emphasises the importance of the ' aium surrounding the cell for the onset of malignant trauslormauon. . As seen from the results of. the investigation, the development and maturaof rlic connective tissue'capsule around the implant and the origination jj lhc tumour go through a series of successive stages which arc indissolubly to each other. Those stages' are similar to the stages oi development of tuntours which Shabad [0] singles out on the basis of conclusions drawn 1252 K. T. J'/.ir.nuv ci.'l A. Kh. Ko';a/: from a large experimental material on the reproduction of tumour, via ,1 agency of various carcinogens. Pour .stage* are mapped out. The first .(up to J5-30 day;) is characterised hy the formation of granulation tj..u which in some areas matures in the usual way and in other area; lead; to abnormal collagen formation and the development of irregularly la(j complexly constructed areas of the capsule. The second stage fup to 3.^ months) is characterized by the formation of diffusely scattered accumulation of connective tissue cells and a pronounced wave of irregular accumulation of mucoproteids containing sulphate and carboxyl groups. The third ;;j. (from 6Ve months onward) is the formation of focal proliferates (prcsarcomijl with increase of the irregular accumulation of acid mucopolysaccharides (mucoproteids). The fourth stage (from 97* months onward) Is the fo.-mniioa of malignant tumours, namely sarcomas. It must be stressed that our-findings of morphological and histoehemica' investigation do not point to any extinction of cellular activity in a latent period at the 5-6th month, as was found by Oppenheimer et al. [14]. R&S 109780 ' SUMMARY , 1. The connective tissue capsule forming around the implant has a noahomogeneous structure. 2. Histochcmical investigation indicates an abnormal course of the pro cesses of collagen formation in the developing capsules. 3. The development around the implant of a connective tissue capsuh its further maturation and the formation of a tumour pass through a serin of stages closely interlinked with each other. 71tnlined by L. Summcmiuo REFERENCES 1. KOGAN, A. Kh.`, Arkh. patol., 2, 63, 1952 2. KOGAN, A. Kh., CHECH CUN, A. S. and ALIEV, M. A.. ArUi. patcI, 2, 55, 1953 3. KOGAN, A. Kh. anJ CHECHCL1N, A. 5., Pat. fiiioL t ektoer. ter., 3, 39, 1957 4. KOGAN, A. Kh,, CHECHCLIN, A. S. c: .7.. Arkh. patol., 1, 44, 1958 5. KOGAN, A. Kh., Pat. thiol, t ektpcr. ter., 2, 74, 1959 6. SHABAD, L. M,, Vojtnik khirurjii ini. t. 1. Grekova, t. 74, p. 6, 1954 7. DANISHEFSKY, L, OPPENHEIMER, E,, V1LLHITE, M., STOUT, A. and F1SHMAS, M,, Cancer Re'., 19, 11, 1234, 1959 S. DRUCKREY. H. ar.J SCHMAHL. D,, Zuchr. Naturforseh., 1152, 76, 353. 9. NOTHDCRFT. H-. Arch. Ge<chwul>tfortch. 9. 101. 1955 10. NOTHOl.'RFT. 1L Strahhntherapic, 1C3. 192, 1956 . ; i .11. OPPENHEIMER. B., OrPENHEIMER, E. and STOUT, A., Science, 113. 305, 1953 ' 12. OPPENHEIMER, B.. OPPENHEIMER. E., DANISHEFSKY, T. end other*. Cancer R*. 15.333,1955 13. OPPENHEIMER, B.. OPPENHEIMER, E. and STOUT, A...Proc. Soc. Esr- ^ 1 a. Med., 67, 33. 194S t4. OPPF.NHF.IMF.R, B., OPPENHEIMER, E., STOUT. A,, ^TLLHlTE, M. and DAM- ; SHEFSKY. I,, Cancer, 11, 1, 204, 1958 i