F/EPA United States Environmental Protection Agency- Office of the Administrator Science Advisory Board Washington DC 20460 SAS-EC-88-040D September 1DR8 Finn.' Report REVISED 24, 1988 Appendix D: Strategies for Health Effects Research Report of the Subcommittee on Health Effects Research Strategies Committee ------- SOT ICE s town sorrittan as a part of activities of tfieScienca Advisory Board, a public advisory providing extramural scientific information advic* to Administrator officials of Agency. The is structured to provide a of scientific matters r«iat«d to facing tht* A<.j<*ney. This for 'by Agency; henc«i contents of this do necessarily poiiciaa of Enviromi«nt«i Prot«ction or of Any of or do or use. ------- U.S. Environmental Protection Agency Science Advisory Board Research Strategies Committee Health Effects Group Chair Dr. David Rail Director National Institute of Environmental Health Sciences 111 Alexander Drive, Bldg. 101 Research Triangle Park, NC 27709 Member Dr. Ejla Bingham Department o£ Environmental Health University of Cincinnati Medical College Kettering Laboratory 3223 Eden Avenue Cincinnati, Ohio 45267 Dr. Bernard Goldstein Chairman, Department of Environmental Community Medicine UMDMJ-Robert Wood Johnson Medical 675 Lane Piscataway, Jersey 08854-5635 Dr. David Hoel Director, Division of Biometry and Risk. National Institute of Environmental Health Sciences Research Triangle Park, North Carolina 27709 Dr. Jerry Hook Vice President, Preclinical Smith, Kline and French Laboratory 709 Swedland King of Prussia, PA 19406 Dr. Philip Landrigan Director, Division of Environmental Occupational Medicine Mt. Sinai of Medicine 1 Levy Place New New York 10029 Dr. Donald Director, Division of Human Risk National Center for Toxicological Jefferson, 72079 ------- Dr.- Frederica Perera School of Pjblic Health Division of Environmental Sciences Columbia University 60 Haven Avenue New York, New York 10032 Dr. E.Len Silberqeld Chief, Toxics Program Environmental Defense Fjnd 1616 P Street, N. W. Room 150 Washington, D. C. 20036 Dr. Arthur Upton Director, Institute o£ Environmental Medicine New York University Medical Center 550 First Avenue New York, New York 10016 Science Advisory Board StafC Dr. C. Richard Cothern Executive Secretary Environmental Protection Agency Science Advisory Board 401 M Street, S. W. Washington, D. C. 20460 (AlOl) Ms. Renee1 Butler StaCC Secretary Environmental Protection Agency Science Advisory Board 401 M Street, S. W. Washington, D. C.. 10460 (AlOl) Ms. Mary Winston Secretary Environmental Protection Agency Advisory Board 401 M Street, S. W. Washington, D. C. 20460 (AlOl) ------- OF THE AL AIM Abstract Chapter 1 Chapter 2 Chapter 3 Chapter 4 Title Environmental Factors and Health Kinds of long-Term Research Research Advances in the Toxicology of Lead Baslc/long-Tera Research with Application to Environmental Health Problems Chapter 5 Population Rtsk/Risk Authgir Arthyr Upton James Pouts Kathryn Mahaffey Marshall Frederlca Pereri Wtlcox David Michael Ho§an Chapter 6 ------- ABSTRACT This to the {both basic and applied) of EPA 1 pro¥ides a historical , the and of of and disease, on the underlying of toxldty with Implications for risk and prevention, and indicates of the Is clearly Inadequate. 2 a distinction the and applitd long-term health effects of EPA by providing specific Illustrate the for "generic" Issues as well as various activities application to specific problems and specific sittings but which be carried on a of years, An has to explain how EPA on baste of the by pirticylarly as it relates to the regulatory Mission of the In 3 the 1s as the to Illustrate the piice of and necessity for long-sustained, basic activity 1n the of a foundation for constructive action 1n in health. Contlnyed Investigations on toxldty are at one and the the justifiable and yet of in the field of A of leadl activities with potential application to environmental are 1n 4, It to highlight activities the in this of of the "new biology* field, as the of and the and yse of to and and for relating to Other In toxicology, and are An of and validation. is as a new find yseful applications in of the Internal stryctures, states, and of biological Finally, In 5 the of of risks 1s pirtlcylarly as if relates to the role of 1n the quantification of possible risks. inclyde of or extrapolation prlmiry or Indirect of ind 1n and of biologically effectlte 1n and analysis are also the ------- -2- of on and within any ire tnd 1s as If the EPA 1s to * policy in the possible. ------- -3- 1 AND Arthur The cerstyfy has the of which caused the morbidity and mortality in previous generitlons. In the of the world, the life hts doubled, now the biblical of and (Figures 1 and 2). This mlriculoys to our great grandfathers, has rtsylted 1n our of the relationship and the broadly These advances, the resulting 1n igricylture, nutrition, sanitation, public health, and all but 1nfectioy$ and parasitic as of in the Industrialized world, afflictions as of 1n the industrialized world tre 1n and chronic and (Figure 3). until largely as or of aging, ire now Increasingly to Qyr 1s to the and to 14). AMD OF OF AN The "environment", broa-iy, all that act on the ind ,-Jdy. of the ire or by himself. They and physical in tir, food, the and the workplace. The "eiwlfoment" 1s and changing. Inevitably, ft a of 1n wd mitt pie the of 1n the of any given may the effect of and the conditions of Air of pollytion, such'as listed in Table 1, to in and mortality. The of ire and may vary, on the in and in the (4), On at relatively in the a variety of ire to Include (e.g., ¥loyl chloride. ------- ,4- radon), metals (e.g., lead, nttrcury, arsenic, nickel) and dusts (e.g., silica, fibers, coal) (5). Also well are the effects of chronic to cigarette The Incidence of lung canctr has risen precipitously, In pinllel with the Increase in cigarette consumption (Figure 4). In smokers, furthermore, there 1s a systematic relationship the of inhaled and mortality lyng cancer {Ftgyrt 5), other cancers, heurt disease, and respiratory dlseasts. • Lesser effects tentatively attributed to passive Inhalation of cigarette in chronically nonsnokers, The ultimate effects of chronic low-level exposyre to otttr widely prevalent and their derlfitlwes (such as sulfur dioxide, nitrogen dioxide, tlpyrene, and various participates) are less well Although the air pollution produced as a result of conbustlon 1s a direct cause of respiratory fatalities, 1s no of their number; however, seven! of the of fatalities ittrlbutable to the combustion of in electricity (where 701 of coil occurs), (81, for S and §00 fatalities result per 1000 Mwe of electric yeir pollut1 on by fired plants, A by 1n this the and fatal 1t1«s per year per Mm of (9,10.1, On the of a value of 7 x 10 Mm of electric 1n the U. S, by the of coal, the Imply that yp to 700,000 fatalities per year nay resylt of in the U. S, Within the uncertainties of this 1t wtll with a Inference by Wilson "50,000 the 2 million who die in the may their lives by »1r pollution" (11). One may question, therefore, the to current i1r standards provide protection against the potential long-term health effects of coal prodycts, which cannot be specified with certainty on the of existing (12), It 1s 1n the pollution nay to health 1n the chronically espectally children. Of Increasing concern Is the to which elevation of the concentrations within and buildings, by weithtr~stripping and other heat-savlnf neasyrcs, the risk of lung 1n (13-15). air-borne pollytints with potential health Includt allergens of varfoys kinds. Although susceptibility to differs widely 1nd1f1dui1s» sliable populations ire it risk (4). The full significance of air-borne as of 1s far established and strongly merits (4). In the third world »1crob1il nation of drinking still ------- -5- s of of pollution no on i in tht of his in the two of 1n the U. S.: and (4,11). It 1s to be polluted in i of {Figure 6), to by metals, toxic agricultural and of chlori'natlon or ozonization. The of of 1n be precisely on the of existing is to the relationships and to clarify the may the (17). is to the "you are you eat". Overall 1s Influenced by the of 1n the diet, the of of (protein, fit, the of the are tht 1n of naturally OP and the of or (18). IB 1s the for and tht for life and In tht of for to the may tht of the 21; tht relative of any of to the of t of to toe (18). In It 1s nay art of or the of tht net of the may the the two of of the to the in the of cincer, and of In tht world, the of strongly As to and at levels his to Collectively, the of and of my by (5). art significant 1n to mty it of In a of is to To the ------- -6- of risk factors, of 1) and quantitative of and In the 2) surveillance and of wort-related and 3) of clinical and laboratory for prior to causing for Identifying high-risk and for at ire readily arrested, or (4). Love Canal and Times to two of (Tables 3 and 4), to the for of toxic it 1s clear deficient in many instances* the of- optimally and cost-effective will research, as will of the magnitude of the risks by prevailing levels of contailnjtlon around existing sites (21-23), The of on alone. This be to of Involving liboratory and be as 1n of the of toxfcoloflcal for 1n the 7). TH1$ will to the In of differences and the Interactive effects of the art characteristically it Of FOR RISK AW As of the of the of of way differently to the total. the of a or of vary, on the of as well as the In directly, act indirectly. the of or on the of {*}, of ttt« of it is difficult or to tht of i of Its of of the and of of i Is also essential in its for on the of Its In of the extrapolation bt tht of (25-26). to tht It not bt for of no art or ------- to exist. Include the «ytag«nic, carcinogenic, ind of the of Ionizing (14) and {4). 1n this connection 1s the evidence to life and eirly Infancy nay impairment In the of the brain, the relationship for which to hitherto considered nontoxlc conceivably no £2? I. In addition. It is not always feasible to el initiate a toxic from the environment, the practical for mitigating Its noxioys effects nay be to or In individuals. For this of the of effects may be crucial, as well as the ability to Identify Indlvldyals it early for effective protective Intervention. for monitoring populations, as well as for the environment, are Any of the of in not the influence of and Influtnces (28}, ts one of the It may affect 1f not all, inflyences, directly or Indirectly, Mortality of the of to inversely with ind (29). The who live In the In of nil nutrition, ind living conditions, to and air pollutants, and to conditions,. ind which on The of dally living in who are not economically al so exercise, of sleep» of smoking, and of of ire In {30), IE (31) and Day {32), who and Is in the at large. Also Is the of and (331, to the of in not to or to The of at ill levels to to the for to the The -- for 300,000 per in the U. S. ind {33) -- the of factors, f ind In the for or for ------- -a- AND OF As environmental an range of illnesses. They Include, for In chronically to acid air pollution, leukemia In to benzene, lung cancer and in Individuals to chronic kidney diseise neurologic impairment In persons to solvents, impairment of brain in children early 1n life to lead, disease in indl¥iduals to and impairment of reproductive function in men and to and certain pesticides. Such Illnesses afflict millions of in the United States. environmental arise from conditions, can be the elimination or of exposures at the source; i.e., prevention. They are also to prevention -- i.e., early detection in can still be controlled or cyred; this however, on efficiently and effectively Identifying populations at high risk. Finally, their nay be by tertiary prevention; i.e., prewention of complications or disability by application of diagnostic and at all requires the of specific and on the and 1n the two are to environmentally-Induced Include, for the Mr Act, the Safe- Drinking Act, the and Act, and the legislation. In of legislation, environmentally-Induced 1n society. Given Illnesses art and highly why do still persist! A of to situation. 1. Despite it two of regyl'atory and scientific ind effort, relatively little 1s the of syr^hetlc chemicals. tnd on a of relatively as and lead, and their Virtually no information 1s available on tht toxldty of 80 of the 48S000 In (Figure ?!, for of which are closely and which most 1s ~ -- reasonably Information on 1s available for only a of (Flgyrt 7), twaluatiort of new 1s inadequate, Z. Physicians art not to tht as i of do not of for to identify in origin of are 10 of (34K In of origin are assigned to is age or and for or ------- are lost. This of Inaccyracy in diagnosis is by the of origin are typically not clinically or pathologically different by lifestyle and factors. 3. Physiciins do not training 1n medicine. Very little is In to physicians in training to the of toxins, or to the associations and disease The medical receives only foyr hoyrs of training in environmental and health dyring the four years of school {341. 4. Persons are typically to one In the environment and do not realize at all. Further, the of environmental conditions only years after onset of during this long latency (Incubation), may may be to i variety of environmental exposures, nay syffer various exposures, and finally my hid All of the difficulty and scientists In to the of Illness. 5. The U. S, and art to and conditions; Unltatlons the of to Inspection and ictlons. 6, unreliable and for environmentally significant of the of of environmentilly Illntss In our society. As a result, the picture not an of the of disease. In a lack of on the toxlclty of the majority of chefflfcals, Insufficient and of physiciam, and surveillance ill efforts to the of 1rt the A plan to the 1 lance, and of 1s fir the detection, and of 1n is Mew York, Mew Jerstj, and California as for an {3SJ. OF the It Is of the of Illness 1n the y, S. 1s wholly or in to Thus, if the billion In the U. S., a 1s on ire or ------- -10- to (36) and are preventable. the of be It is enormous. 1n the light of the of illness to tht U. S. popylition, the on to Illness are relatively snail. In for only of the 15,121,55? RIO by HIH specifically to on (37). This sum to 0.251 of the of In the U. S, year {371. The sun for the by ill was fir (37}, in of EPA's to the U. S. It 1s clear the and for the to be The In life resylt 1n the Influence of Defined broadly,, §11 act on the and Inflyences, are and in «1r, the and the of are by man to his at levels, nay it levels, t In tht §n tht of of In or ifi as of the of any one In tht of a be By the 1t 1s difficult to tht to may a at any pirticulir level. In of of is 1s and be on in or To our of tht of in priority be to on the following: 1) and of the 21 of and rates, to the of specific to be to 3) of for of to and for identifying 4} of for tht activity of and it low and In 5) 1n for particular to and and 6) of the of is for 1n and In the and of In ------- -11- »1goroys should be the application of existing 1) public and professional education, 2) standards-setting, 3) of new and existing legislation, 4! law and 5) research to evaluate the efficacy of measures. In pursuit of Its Mission EPA in coordination with other agencies and institutions a long-range strategy addressing of the ------- -12- REFERENCES 1. Jones, H.B. A Consideration of the Aging and Life Unfvi"rsT% of (TTTf^nTa~,"TTad1at1orr[.aboratory,~~~ ~» 1955. Fries, J.F. Aging, Mitural Death, and the of Mortality. He* Engl and_J .Jted . 303 : 1 30- 1 35 , Oonabedian, A. » Axelrod, S.J. , Swearlngen, C. , and J. Mt£i_£aT_ Care _ Chart . 5th Edition. of Public Economics, U n T ¥ e rs T ty o f Hi c h i ga n , Ann Arbor, Michigan, 1972. Second Task Force for PI inning in Science. Huma n__ Heaj_t h a n d _th e_ _En_v i roniB_en t - re h U. S. Depafliiffr¥rWiTWrTa5c¥€Ton7^ni~W NIH 77-127, MiShlngton, 0. C., 19??. Levy, B.S. and O.H. {Editors! _Qcc u pa 1 1 o n a 1 He a 1 1 h : andPrev en t in g^ wo rtt - Re Ute d D 1 je a s e . I 1983. ~~~ ~" 6. Cairns, J- The Problen. ScU^Amer. ^33:64-78, 1975, ?, Doll, R. An Ep1de«lQlG§1ca! of the of 8. H. of EnyP^gctlon, . ~~~' 9. Morgan, M.6. , Norrls, S.C. , Htnrlon, M. , , D.A.L. » tnd Rlsh, W.R. Technical 1n Policy - A Sulfur Air Pollution ^:201-Zli, 10. Morgan, M.G. , Henri on, H. , Morris, S.C. , and , D.A.L. uncertainty in Risk Enyr1 ron ._^ Sc 1 . _ Tech no 1 . 19: , 1985, 11. to the Editor. Public 238:10-11, 12. Ttsk on and and Cancer and and Lung P*_seasg,__3rdi to C~TivTroSienlaT^ Igency7~~ [[[ ' ............................... ..... ~ ....... ~ D. C. , 13. on Pollutants. Indoor .PoVlutants^. of o. c. , iwn 14. on the Blologlctl of ------- -13- 15. Council on and Evaluation of Occupational and Environmental to and Rtdort"™^ on~Ra^TIfTonnjro^eTtfoirTf»3~Heasupements , Washington. 0. C, , 1§84. 18, Dlckson, 0, Toxic on Clean Up.. H:46-5°i 1982* 17. on Drinking Drinking and Vol. 4. National of Sciences, ~~~~~^ 18, on Diet, Nutrition, and Cancer, Dl^et,- Nutrition, and Cancer. National of OTTT 19, Doll, R, The Epidemiology of Cancer, In: 1n Cancer jyT^^ Philadelphia, 1979, pp. 103-121. 20. Wiiss, 8. and Clarkson, T, Toxic and the Implications of for Technology Transfer. jjjTban 6£:216-24Q, 21. Office of Tectin§1ogy U, S. CoigressTwil^ 22. Materials of TJasRTrTgton, 0. C. , Ii83. 23. §n to of Mul Contaml of 24. Nation*! of - Council, Tox1c1ty NaWoniT" 25, on ~™^^ 26. i.S. to Teratogentsfs, Effects. j._^er.CoTJj_Tox1_coV._ ^;113-123S 27. Bellinger, 0. , Levlton, A., C. » H. , and H. of and and Early N, Engl . J. 316:1037-1043, ------- -14- 28. H. §fw™!!t^^ 29. Mortality: and Wales, 1970-1972. 1978. 30. Belloc, N.B. Relationship of Practices and Mortality, Prev. Med. 2_:67-81( 1973. ~^ 31. Lyon, J.L. J.W., and West, D.U, in and non-Momons In During 1967-1975. J-Nat-Caricer Inist. 6:10SS-]Qi2, — __ _ 32, Phillips, R.L. , Garfinkel, L, » J.M. , M.L., Lotz, T, , and Brin, B, Mortality California for Sites. J. Mat1.Cjncerlnst. 65_:1097-1108, 33, General, ^okjjiqajidjjealth. of Health, Education, ind ............ IT. ........ T. ....... , ........... TfTf . 34. Levy, 8.S. The of Health 1n United fl¥e-Year Follow-Up of An Initial Survey. :79-80' ~™~ 35. S.B. and P.J. 1n Mw of °~~~~~~~~ " 36. of ___ . 37. national Institutes of Health. U, S. Printing Office, Washington, 0. C. , ------- -15- 1. Atr Pollution of P1ace Valley, 63 Oonora, Pennsylvania 20 London, Mew York, Mew 200 London, England 700 ------- -IS- 2. In May of 1, By providing of §r -- of -- of or or -- Products of -- of residues) 2. By affecting of — Pro¥lsf§n of for of (secondary amines, nitrates, nitrites) Inhibition of of as tn (Vitamin C) -- Alteration of of bile and ( — Alteration of of by fat, In -- of -- Affect on of (fits, 3. By of — (alcohol, -- on In (fiber) Inhibition of A, 191 ------- -17- 3. Pollution P8B« St. Louis, 1973 Lead 1976 Italy 1976 California 1977 Love Cinal, Mew York 1978 Oioxin Oioxin New aPC8 as P88 a$ bfphenyls, as and it 1 ------- •18- Table 4. Examples of Outbreaks of Mass Human Poisoning From Toxic Chemicals Date Location 1930 1934 1952 1952 1952 !955 1956 1958 1959 1960 1964 1967 1§68 1971 1976 1981 1984 U.S.A. Detroit London Japan Morloga (Japan) (Japan) Turkey Kerala ( India) Morocco Iraq (Japan) Qatar Japan Iraq Pakistan Spain BNopal TrJorthocresyl Lead A1r pollutants Parathlon Arsenic Methylmercury Hexachl Parathlon Tr1 orthocresyl Ethyl mercury Methyl mercury Endffn Polychlorlnated bfpttinyls Malathlon Toxic oil 01 Mo_._A£fecte_d 18,000 4,000 4,000 1 ,800 12,159° 1,000 4,000 828 10,000 1,022 646 691 1.665 50,000 12.SM 2»000C of Thest tht of It was that and 131 died. C0eaths« Ttte full of lingering and unknown. (Froii 20) ------- -19- m 40 so OF ao figyre 1 1n and 1). ------- -20- 100 2 The Survival In the U.S. 80 of the the 1900 and the hid by 1$ n§w the of in 11ft. 2). ------- -21- fff all 1 flf fttHJ 221 AN Of P"^™^^ * Mrty C of c — MIIUU> ^^ 3 a a - 3 a 3 of 1rt the !S67» as 3). ------- -22- in ind 1n ind 61. ------- -23- for «§• mm 2M tm t t t I » at ^^^^^^^^S iBUPii • Sit 1 4t Figyre 5 of In In to of Per Day. 71. ------- ,24- = pollution - may not be but the 1s not 0 metals, is zinc, and i development * tnd and indystrlal Figure 6 (As by U, S. 161 ------- -25- :=>V t»f «MI M A II I* 1 M tl I U 4 J* 14 U II u 1 * = line » toxlclty » ) - No of on of Different for 241. ------- -26- Chipter 2 Of LONG-TERM Foots LONG-TERM HEALTH EFFECTS SUPPORTIVE OF EPA I. Basic Research Bask research in EPA programs nay or nay not be directed spedficalif at support of certain applied research programs. Such basic research miy seek, only to understand detptr levels of the general universe of problems attacked in the specific, discrete long-tern, applied researches {such as described just below). The general basic research philosophy 1s that undtrstandlng the chtuticali disease can lead to earlier detection or better for health effects (and better designs of epidemiology studies), analytical » etc. All of this can and often lead to for regulation and, thus, regulation. Section 111 below) of this basic research cm be it using of the "new biology" to our ability to or to identify and quantify specific effects of tor had actors In) nlxturts of chenricals occurring "naturally". Ov«n11 the " distinguishing of b«s1c 1s 1t "generic" Issues, ind 1t not necessarily be any on« specific nor "qylcfc* A$ 1t be for seven! years to be effective and to glvt tht kinds of findings will be uscfyl to "applied" It 1s, true that often the useful ftcts and new in resolving tny environmental turning to laboratories basic research. There art of the kind of (and 1$ risky) any of the of night be; A. New to ind quantify dioxlns lisle his Identified characterized in intracellular "receptor" for 41oxin$ tn^ related curled out rtsylted 1n tht partial purification of this receptor, ami of the ind specificity of severil of ttw b1§1o§1ci1 of d1on1ns. Recently, the "ntw biolofy* ttchn1fu«»s this "dloxin* rectptor his clontd and now be available to (and In his to one posslblt application of use of this to tnd of it lt«tt d1ox1n»Uke 1n wlxtures—ptrtlcularly of the dloxlns 1n soils, Ttii on tht *i1ox1n* 1s tut kind of effort «iy now to "fruition" (e.g.. 1n tt« nw ------- -27- for dloEfns In mixtures). 3ut it his over years, and to the practical/applied of Gbjectlrts, hts not of to of the EPA B, New for detecting to toxic chenical s The are a of steroid. 11p1d, and xtnobiotic mftabolizing 1n a wiriety of living (from yeast to . over at least 35 his led to of the diversity and of in The "new biology" his given y$ new tools for qyantlfylng and Identifying of It is now possible to "fingerprint" the an^ of of P-4SO in of (Including and plants, research his described 1n detail of P-450s to various '(including all long-continuing, bislc be giving us for looking it the of plants, animals, and to chemicals--e.g., the and of P-450s to reflect to things like Fyrthtr, basic ! ly 1n nay give ys a for and (of to for flc P-45Qs. II. irt of activity to and but be on a of can be 3 U ind pirts/step$--whert one usually be be initiated/planned, 2J a but pirts of can be on 3) and validation. A. 1n This 1s a of of in projects--either In their §r ire but the key 1n Is 1s i and up the 1, The layer and Is i his for Tht ind of ire of tht 1n U¥ in^ on nay be ------- -28- 1 1fe- threaten! ng, direct. in this overall i. The first at the Issue— Is any evidence we ire actually losing The to this {data supporting this) 1s still and (at in quarters), but the first indications that to a loss; therefore., 2 b. The to be: be this loss of Is any contribution (e.g., chetical) can destroy and is likely to get to the layer! about the chemistry md Interactions of light, *nd had to be first. arc still out at this c. The was to gather the of materials/chemical s (e.g., 1n the liyers of the New for collection of etc. to be developed, validated and d. real-life of had to be and for their contributions to the problem. i, as to bt in the of it the had to be Thus, of here-— chemistry, sociological/psychological , §nd political. the to be In the overall of dealing on the of just and on of preceding, 2, He and of rain A of but ill or of pollutlon has the effects, and really r§1n 1s to be primarily Industrial art ind of One In Is the to flora, and 1n 1s due to and or 1s by cars/traffic, A of has and art continuing. It fs of the the Is "yes" to effects/hunm rain and car This i of and out One of the on all f Including of of 1s the In if at an it ------- -29- 1n 1987. The of this will be in |nv1rogient_i1_ HtiUh Perspectives 1n This effort In bo theology""and ' TuMarTheTrtrriFfeTts of rain his on for years/decades, and ire only now barely visible. B. Long-term studies with concyrrent arc Just a time—the are the study just can't be in short "pyrely" epidemiology fall hire—where the effects of low level, chronic or to resolting only years after exposure or In populations that "age" to effects. Most studies on possible of cancer or on carcinogenic of chemicals art So are i¥a1uit1ons of the of other slowly developing effects/diseases (e.g., kidney failures, liver and CVS or CMS effects). evaluations. involve multiple at the but continuing for a long on the populations. Chronic toxldty In animals Are a of this kind of There are i. The "Six dttes of of air pollution—comparing various indices of 1n H¥in§ in 6 cities of widely ¥iry1ng of pollution. This has on for now, part of the Increasing clarity In this resylts from now 10 years, but 1s the of new and analysis to the for effects. The point 1s this of the populations/regions years to and to clearly the In air pollution fwhlch the of the in 6 "regions". The principal effects now are on the (lyng fynctlon but ft.g. „ kidney, C¥S) my be to be as studies continue. 2. The effects of polychlorinated biphenyl (PC8) on childhood This with in the U.S. and (e.g., oil 1n and Japan ind Hke the of oil with PCBs along highways, and the of living near, or walking along highways). From short- and 1t was of PCBs not acytely but in and sybtle. Therefore, to follow {for 1n of and especially in any children The of levels of to on childhood ire now In only dentally-exposed populations and a large of "Itss-txposed* and "nomaF/unexposed and children for 3. The of Again, an accident—the of and the of this and of the 1n are still ------- for effects, again, are and subtle. C. and validation of In the for detecting and quantifying new environmental toxins/problem do not it the "problem" ire first discovered. This set of "long-tern" activities Is vital in any to and affect environmental hazards, are but only a few be given; 1. Dioxins fPCODs) and (PCDFs) for ind quantifying "families" of did not the first "poisoning" in The of materials 1n Is small, and yet» in animals, chemicals toxic effects at low concentrations. We are only now the to quantify and selectively identify and the of chemicals found 1n real life of the In analytical to this of The of and analytical to Identify the dlbenzofyrans is of the and dloxln and also as contributors to of the toxfcologlcal This his it and 1s not yet. Validation of all 1s still occurring. Z. Lead W1th/1n we of the toxic 1n "deep" tlssyes. at without painful or the yse of 1$ a If the of 1nfomat1on *e are to be generated—particularly for long-term stydies, or for chronic {although this infomitlon miy be vital for metals, to only briefly in tissues and flyids. of and In relatively like (or fat, etc,), to of are Non>1rwas1ve §re especially useful/attractive for for In the of lead be now X-ray flioroscopy. Validation of this 1s now taking place—total to use will be ------- -31- ten years 1 f all well — a long-tem effort typical of several others. III. How EPA on by Federal Agencies Health within the EPA Is ultimately directed the regulatory mission of the Agency, While 1s often of an "applied" and/or "immediate" which specific the with in an manner, or fundamental research is the only of Improving the scientific rationale underlying regulatory decisions. It is vital the EPA scientific staff maintain current of re1e¥tnt by performing research within the Health Effects Laboratory and by closely following the latest 1n toxicologies! The effectively Its mission without scientists 1n of tools of basic research. Without this and health scientists within the Agency would be unablt to effectiwely translate the findings of fundamental into the sypportive of the Agency's regulatory mission. by EPA represents only a fraction of which 1s to its regulatory mission, the rely heavily on information by Federal particularly by the various Institutes of the of Health and Services, These organizations for of the scientific In biology, and research that have of for monitoring, dosimetry, to*1colog1cal testing, and Basic at the national Institutes of Health has substantially the Agency's regulatory and policies. on the of mutation, xenobiotlc metabolism, pnarmacoklnetlcs, and moltcular performed at the Institute of Environmental has applications it EPA In and metabolic activation for Iji vitro for ng, for biochemical epidemlilogj. by the Institute on of cardnogenesls and survelllanct has contributed directly to the of toxtcologlcal and gyidelints for by the EPA Office of Health and EPA 1s directly widtly and in the of neurotoxlcology. The discovery of cell within the their differential vulnerability! is Itadlng to in of of neyrotoxiclty and for the of neyrotoxicologic will to future guldtllnes for neurotoxiclty testing. In addition to the yse which the of basic information by indirect ------- -32- (infomation 1n the literature and discussed at scientific forums), the also active collaborations which of findings and/or expertise. EPft scientists frequently in collaborative studies with scientists in as well as their colleagyes in who nay be by agencies. These research efforts often of expertise in new technologies and new findings that may applications to the regulatory mission of the Agency. As an example, research on mechanisms involved in the successful fertilization of the oocyte has led to int.^ragency collaborative to improve for the evaluation of maie fertility. Other efforts delineating the fyndamental factors involved in absorption led to joint interagency projects on the of methodologies for the of the kinetics of exposure. Clearly, it woyld be possible to this list of relevant since mych of the scientific information utilized by the for regulatory decision-making guidelines fomylatfon rests on a foundation of basic research. ------- -33- Chapter 3 IN THE OF LEAD Kathryn PREAMBLE The place of and for long-systained basic research activity In the of a foundation for constructive action in important problems in environmental health coyld be Illustrated by reference to any of several current He the and its dangers or toxlclty to this purpose. as § public health problem has for Iff not centuries). Yet howt what, and to do Its effects and treating not have only recently, and only as a result of long-continuing research. For one thing, only long-range, nwltidisciplinary, continuing basic has giwen us the varied tools we to of the subtle (yet important! effects of lead. We counting to worrying things like behavior and tn children—but only we now for of lead. This 1s the story of an health only slow-n»v1ng to explain] and by far-sighted who long-ringe was and would to be cost-benefit Background Understanding the of by exposure has 1n this centyry. Into the toxic effects of a his the discipline of clinical and toxicology for the five Fundamental mul ^disciplinary laboratory 1n as biochemistry ana'physiology his a najor key to this progress, has long to be acutely toxic at exposure. In addition, *e OB 1n tht 1970*s and two of of on it levels ont of the of the on tht Is in of cularly In children, to lead, as 1ngest1on of paint, his to a characterized by convulsions, and 1n In with to lead, a "wrist and drop," and art the conseqyences. ------- -34- The challenge has tc ".arstand that the of health by lead was e-.rns1ve the clinically-obvious has this challengi esisdally difficult Is lead pollution has at lewels, an byrden of 1n a sizable portion of the popylatlon. During the 1970's In metropolitan areas, young children frequently Nad concentrations greater than 40 g/dl; a concentration associated with several neuropsychologlcal The Is to the etiology and severity of health are so are considerad "normal." Irs the of public and medicine mortality iftd morbidity (i.e., reports) to understanding the This reflects and has been possible only of lonf-ranfe support of enflronnentil research. exciting recent findings with to understtndlng of the toxicology of is the realization 1s of producing toxic effects in adults and children at relatively low levels of I.e., levels that ire insufficient clinical Only a igo of "sife*. 1s now to a of • toxldty. h«$ of Is a Involving Th« red cells, the and -the are the In In early IfOO's so routinely for (1935) the of to poisoning for the and 1133 wts in of 3400. The of children* nho art sysceptible to the of In the 194.0's the 1%0's of of the the Prior to the of hi4 « rate of §51 of poisoning 1s the predominant, :*For and (1143) and clinicians of poisoning sensory deficits, and bthavloral dysfunctions. Ptrlsteln and (1966) 1n 371 of who poisoning withoyt of cncephiloptthy, Through to identify and to of to clinical of toxicity a control; not clinical of intoxication, identified «h!ch ire it ------- -35- hlgh-dose exposures. The limited reversibility or 1r reversibility his documented fn many of the clinically-reported, neurologic effects. Using these clinical studies as a gylde, long-range, multi disciplinary reseirch has extended the understanding of lead toxicity .to the cyrrent emphasis on btomarkers of exposure, dose- response relationships for specific effects, and identification of sysceptible subgroups for these effects, ResearchF ind i n gThe9 7 0*sinM9B(rs The general picture of adylt and pediatrlc lead poisoning has changed in recent decades. The overall pattern is Identification of significant adverse health effects at progressively lower exposures. These can be arbitrarily separated Into neyrobehifloril , hematopoletlc, renil /endocrine, and reproductive effects. As a part of this effort, differential sensitivity of various sybpopylatlons has revealed. Identification of effects occurring at environmental exposyres considered "normal1* has coincided with redycing environmental exposures 'to lead. Only through reduced exposures can the resylts given by toxicology and epidemiology research be evaluated in general hyraan populations. I. Neyrobehavloral Effects Recognition that neurobehavloral effects in children are produced by lead exposures considered "normal" 1n eirlitr (e.g., blood lead concentrations of 20-50 g/dl ) has tht significant findings in the 1970' s and s. Long1tud1nil studies during the 10-15 years built ypon early and cross-sectional studies. The longitudinal prospective designs hate permitted gathering liproved information on exposure histories. Information on exposyre levels and patterns is clearly important 1n assessing effects of a cynulitlve toxicant on endpoints such as neyrobehavlonl function that nay reflect changes induced at far earlier, but critical, periods. The most consistent finding of the prospective studies is that an association exists between low- level lead exposures during developmental periods (especially prenatal ly) ind later deficits 1n neurobehavloral ptrformance. This latter 1s reflected by Indices is the Bay ley Mental Development Index, a well-standardized for Infant Intelligence, Blood Itad concentrations of 10- IS g/dl constitute a level of concern for effects (EPA, 1986). In addition, Impaired neyrophyslologlcal function has issociated with Increasing blood concentritions children. These functional deficits Include changes 1n the auditory brainstem evoked potentials and evidence of lead-related hearing acuity {Robinson et al.t 1985, 1987), 'These subcHnlcal toxic effects of on the central nervous system are generally considered to be peminent and Irreversible, and they are associated with permanent loss of Intelligence ind irreversible alteration 1n patterns of behavior. Belli et il (1987) significantly on tht of tn uppcr-«1ddlt class Itad Itvels 1n the of 10-25 g/dl, adult It veil 10 ind 12 g/dl on ttit 11 ------- -36- for ttw tt i1f It be art one of the of the In tht II The is by Typically, adults ire likely to nervous effects. In the early 1980's investigators to call to "subcllnlcal" by in velocity in lead not neurological involvement et al., 1165). In the 197Q*s et al. 11972, 1975} the slowing of the velocity of the and ylnar md 1n 70 g/dl. Investigations of the of an threshold, coordination, and physiological and psychological 1n with blood below' 80 g/dl et al., 19751, II, his i of clinical poisoning in children and adults. of 1s now to In children at of 30 g/dl. of and the of it i of In the red cells the to of 1n children. The for In 1s with a of 15-18 tt al., 1n to The accumulation of IX as or as protoporphfrirt in Is not only in of but signals Injury, The final of In the injyry to the mitochondria can a of subcellylar and Implications of include; of to all cellylar of calcium metabolism; In control; detoxification of and of (e.g., of 111. 1n children a with glucosurfa, and to of the to in has tnd in adults. adults, in for all ill (specifically, of the liver, and lungsj, nephritis, ind (I.e., dye to not ------- -37- in a longitydlnal of in battery plants tnd lead (Cooper, 1985}. A statistically-significant his In systolic tnd and in old, the 11 populitlon (Plrkle et *1, Impairment of the endocrine functions of the kidney reported to occur at of effects required development of several of research; A. Understanding the metabolic activation of Vitamin D to 1,25-dihydroxyv1tam1n 0. This metabolite is critical to regulation of calcium Ism. B. Recognition that"lead Impairs various 1n biosynthesis and function of 1,25-diHydroxy¥ttam1n 0. Currently, the studied at is the tubule of the kidney. 25-hydfQKjf¥itimin 0, 1n liver 0, a hydroxylation Is by the 1, ,2S-hydroxy¥lt«ft1n D hydroxylase. in vitro (following In vivo of to 1 iTEsTfhi activity of this Findings a clinical Investigation children 1 D lt¥els in to to of Itad, 1n of 1,25-d1hydroxy¥itaffl1n 0 yp to levels similar to 1n children serving as controls et al., 19801. has 1,25-d1hydroxy¥lta«1n 0 with i of concentritions, 12 to 120 et al., 1982b). IV. Early 1n the a of of on with included still-birth rite, and a higher, and early childhood mortality children of high, I§ng1tyd1nal, to of lead, important on effects at the of levels. et al. (1986) the of deliveries the 37th of significantly to at delivery, excluded, the of the The of deli¥ery a* levels In of 14 j/dl or MIS 8.7 the it yp to 8 g/dl. 1n age at by et §1, (1986), et al. I1984J, et §1. (1982), and tt al. I1987a, ------- -38- b). The from Indicate for 10 g/dl 1n 58 and 601 on the age of the The of et al. in in miscarriages and still births In the high-lead In contrast, this concentration was lower for still births for 11 we births. to an 1n the of it low 1s of for the fields of and public health. Until recently, of 25 g/dl and and only five ago the for (CDC) 25 a level of in children. on the basis of it 1s In children at levels this guideline. Thus, the toxlclty of lead it low is to § re-evalyatlon of for the of to The of the tnat in the the and Survty In 9.11 of all children In the - 1,5 million - hid of 25 or tt al., 1982a). children the of (high wi$ findings on the of (high blood In the or» subcllnlcal toxlclty, a of chill ing significance. findings 9% of ill in nation, and 251 of minority children, may be suffering irreversible neurologic, Intellectyal , and ts the result of chronic, to lead. The impl Nations of for public and environmental ire This has a one of the and It 1s t eontinyes the findings ind implications. It will only 1f the of us to are ind research Invtstigitlons on toxlclty art it one and the the justifiable and of activities In the entire field of So has 1n In no it of all the his tet His 1n has off and Is still It Is "apology" for we can for the field of health science, may be the specific of of for any one ------- -39- REFERENCES 1. Bellinger D.C,, Neddlewan H.L, Leviton A., Waternayx C., iablnowltz H.B., Nichols M.L. (1984). Early Sensory-Motor Development and Prenatal Exposure to Lead. Neurobehav Toxlcol Teritol 6:387-402. 2, Bellinger D.C., Leviton A., Witernaux C,, Neddleman H.L., Rabinowltz M.8. (19875, Longitudinal Analyses of Prenatal and Postnatal Lead Exposure and Early Cognitive Development. New England Journal of Medicine 316:1037-1043. 3. Bornschein R.L., Syccop P.A., Dietrich K.N. , Krafft K., Sroti J., Mitchell T. , Berger 0., P. B. (1987a). Prenatal-Lead Exposure and Pregnancy Outcomes in the Cincinnati Lead Study. In: Linberg S.E., Hutchinson T.C., eds. Internationil Conference: Heavy Metals in the Environment, V 1: September; New Origins, LA: Edinburgh, United Kingdom: CEP Consultants, Ltd., pp. 156-158. 4, BornscNein R.L., Grote J, , Mitchell T.. Succop L., Shukli R. (1987b). Effects of Prenatal and Postnatal Lead Exposure -on Fetal Maturation and Postnatal Growth. In:'Smith M., Grant L.D., A., eds. Lead Exposure and Child Development; An International Lancaster, United Kingdom: KTP Press. 5, Byers R.K., Lord E.E. (1943). Effects of Poisoning on • Mental Development. Am J 01s Child 66:471-483.' 6. Cooper W.C., Hong 0., Khelfets L. 119851. Mortality 1n of Lead Battery Plants and Lead Prodyctlon Plants, 1947-1980, Sctnd J Environ Health 11:331-345. 7. Dietrich K.M., Krafft K.M., Bier M., P.A., Berger 0., Bornschein R.L. (1986). Early Effects of Fetil Exposure: Neurobehavioral Findings at 6 'Months. Int J. Biosoc. Res. 8:151-168, 8. U. S. Environmental Protection (1986). A1r Quality Criteria for Leid. Research "Mingle Park, NC: Office of Health and Environmental As$issa«nt,t Envlronnental Criteria ind Office: EPA No, EPA-iOO/8-83/02iif-df, 4», Availible from: MTIS, Springfield, VA: PB87-142378, 9. Hoffman F.L. (1935). Liad Poisoning Statistics for 1933, Am J Public Health 25(2; 1190-100. 10, Mahaffty K.R.» Annest J.L. , J., Murphy R.S, (1982a). «§t1ona1 Estlmetes of Levels. United States, 1978-1980. New Engl J Med 307:573-579. 11. Mahaffey K.R. , J.F. , R.W., Peeler J.T., c.M., H.F. U992b). Association-Between Concentration and 1,25-d1hydroxycholeco1c1ferol Ltvels 1n Children, to J CHn Nytr 35:1327-1331. ------- -40- 12. A,J., Vlipani G.V., E.F,» P.A., Clark P.O. 11986). The Port Plrle Cohort and J Epidemiology 40:18-25, 13. M.R. , A.t S.J. , A,, I.M.. Maconesplc H., R. , Low A, {19821, of and Infant Exposure In Glasgow. Med J 27:113-122, 14. National Council (1972). Airborne in Perspective. Washington, O.C. on Medical and Biological Effects of Atmospheric Pollutants, 15. Perlstfiln M.A., i, (1966). of In Children. Clin 16. PlomelH S., C.» Zyllow D,, Curnn A.. B. {1982}. Threshold for to Synthesis 1n Children, Proc Natl Sci USA 79;3335-333i. 17. fHrkle J.L., J.t Landis J.i., and W.R. Thi Relationship Ltvels and and Its Cirdiovascyltr Risk Implications. Aw Jr of 121:246-258, 18. J.D., B.B., Nicholson J. (1975). of to y. S. of Health, and Welfare. for and 0, C. 19. G. , S., D., C.t S. i., P.» O.A. (1985). Effects of Low to on Auditory Potentials 1n Children, WHO Office for pp. 177-182. 3). 20. G.S., R.H. , R.L., O.A, (1987). of Environmental on the as by tnt and In Children, In: S.E., T.C., in the Environment, VI: New Orleans, LA: CEP Ltd., pp. 223-225. 21. J.F., R.y., A., H.F.. K.R. (1980). In 1,25-dlhdroxyvltatftlri D In Children Lead New Enfl J Wed 302:1128-1131. 22. A.M., S. (1972). for Sybcl1r»1cal ir J Ind ted 23. A.M., Tola S.» S., S, (1975). Subcllnlcal it "Sift" of Environ 30:180-183. ------- -41- 24. T., Ferrari E., Colucci d'A.C. (1965). Velocita de Conduzione Net Satyrnlni. Folia Med. (Napoli) ------- -42- 4 WITH TO PREAMBLE In this a of of the basic/long-term applications to environmental (especially 1n humans). This not the of will or be of to It Is, , however, in at carefyl choices of studies which for the of this their applications could on health. As then, this is a look at the and of has 1n the ------- -43- Of BY INTRODUCTION are cellular are and initially as the transduced of trinsfoming retrovirysts (1). studies can be to causing by of retrovlril involvenent (2-4). Include or DMA as or amplifications. The actlvition of by 1n levels of expression of protein product, or 1n or levels of of in protein, Of The activation of In and and 1n has 1n the In for Imply of are by and this is in fri Induction (5-6K -Alternatively, of and is 1n (7-9), The of bt 1n the of 1s at The low of at two r«s and «re for the of celTTTji (10), in to the of the of specific is be t 1n (ID, The of 1n win bt 1n of of to The of rt$ to one 1n the nwltistep ~of~carc1nogenes1s for a variety of and (5,61, The activation of by point 1s an early 1n and may be the "Initiation" 1n Thus, a by of ras_ cm potentially §ne of the 1n to the cNeufcal. Is this to classify the as a rajj In {6}. The of !n the of Is uncltir at is will as the non-jras 1n can be by or radiat1on~T6K ------- -44- are classified as potentially hazardous to on the basis of long-term caret s stydies In rodents. While rodent tarcinogenesis stydies are often designed to tninic the route of exposure in environment or workplace, the of a given chemical is usually higher than that which actually occurs in human exposure. Coupled. with the appearance of species- and strain-specific spontaneously occurring in vehicle-treated rodents, this complicates the extrapolation of rodent carcinogenic to risk. analysis of tymors from spontaneous origin and from long-term carcinogenesis studies should help the mechanisms of formation at a molecular level. For instance, the finding of actiwating mutations in different of the H-ras_ in furan-induced liver versus finding activating mutations in only one of the H-ras in Hver that the chemical Itself activated the H-ras proto-oncogine by a genotoxic event (12). In general, comparison of patterns of activation in spontaneous versus chemically-induced rodent tumor?, with cytotoxic information, should be helpful In determining the chemical in question is nutageniCj cytotoxic, has a receptor of promotion, or combination of (and of action. This type of anilysls might be of particular Importance for such as furan furfural 112,13) which negative for rnytagenlcity in short-term bioassays. APPLICATION TO OF Another which should be helpful 1n species-to-species extrapolation of risk from carcinogenic is to examine oncogens activation and expression in tymors from different by the agent. For example, K-ras with the activating lesion In 12 were In boWrat §nd Induced by titranttromethane {14}. Even little 1s the ONA properties of this , this is acting in the to 1rt nts and mice. The role of chemicals and radiation in the activation of by amplification, translocatlon, and other sis which can alter expression, is currently Investigated by groups. Also, as life increases, it to chemical-Induced of the progression of to malignant These and similar to explore the by which in may of the uncertainty In risk analysis of carcinogenic ------- -45- REFEREMCES 1. JM. Viral Cell; 42:23-38. 2. Varmus HE. 1984. The Molecular Genetics of Cellular Annual Rev _18:553-612. 3, Weinberg RA, 1985, The Action of in the Cytoplasm and Nucleus. Science; 23Q;?7Q-77i, 4. Bishop JM. 1987. The Moleuclar Genetics of Cancer, Science; 235:305-311. 5, Baroacid M. 1987, las Ann Rev of 56, in press. 6, M, Reynolds S. Activation of by Chemical Carcinogens in: The Pathology of Neoplasia. A Sirica, ed, Plenum Press, N.Y., N.Y. {In 1988J. 7. GM, 1C, M, HE, JM. Amplification of i-myc in Correlated with Science 224:1121-1124. 8. RC, GM, Sather H, Dal ton A, SE, KY, D, 1985, Association of Multiple of the With Progression of Neuroblasts, The New Journal of 313:1111-1116. 9. OJ» Clark GM, SG» Levin WJ, Ullrich A, HL. 198?, Breast Cancer; Correlition of Relapse and Survival With Amplification of the 235:117-182. 10. Land H, Parada IF, Weinberg RA. 1983. Tuntorigenie Conversion of Primary Embryoflbroblasts Reqyires at Two Cooperating Nature (London) 304:596-602. 11. Barrett JC, M, Koi M, 198?. Role of and Tymor Sypressant 1n a Multistep of Cardnogenesls, In: Syraposiyin on fundanental Cancer Research. Volume 38 (F. Becker, ed.,), in press. 12. SH, Stovers SJ, Patterson R, RR, SA, MM. 1987, in B6C371 Liver Tymors: Implications for Risk 237:1309-1316. 13. Tennant RW, Margolin BH, Shelby MD, Zeiger E, JK, Spalding J, Caspary W, Resnick M, Stasiewica S, B, Minor R. 1987. Prediction of Carcinogenicity in fran ln_ Vjj£0 Toxicity Assays, 236:933-941. 14. SJ, Glover PL, LR, RR, SH, MW. 1987. Activation of the K-ris 1n Kit and Lung Tynors Inductd by Chronic to Tetranitrotftethane, Res 47:3212-3219. ------- -46- AMD Fredertct P. INTRODUCTION in in molecylir biology and biochemistry the of to of to carcinogens. highly techniques can and the internal of carcinogens (the of the carcinogen or Its ieTaSoWtFTFF and fluids) or the b1_oT£g1cainy_ effect 1ve_ {the has with eel TlTTaTiSaTn^TeTilTCrTucTrTs ON A, IN* or protein) in or a This of could be especially valuable in of etiology by providing a mechanistically relevant link on the one and clinical on the molecular in and the potential to of risk in to of a carcinogenic to applications of "adduces research" could directly at EPA for of and risk to Various are available to Ch«i1c4l-8pec1f1c lesions (such as for ONA and as well as specific biologic alterations (such is or cell mutations). -Table 1 jives of currently available for the biologically of As c§n readily be all pertain to genetic toxlclty. Moreover, all available on readily es for the itself, limitations, biological significant potential In etiology and risk ------- -47- Tabl« 1. of Biologic Methods End Point F1jd Biologically affective ing, fluor- HBC Spectrottetry (protein) RBC analysis, HPLC, gas chroma togriphy Excised HPLC, UDS Oil cylture, • WBC SCE WBC Mcronyclil WEC rations cell Autorid1o§riphyf light MBC Somatic, cell RBC tglycophorin A) quality Analyses of count, motlHty See 1 fas cells; cells; DMA HPLC»Hlgh ------- -48- and the of 1n the 5 and Illustrate a of and limitations to biological 1n (2,3). The biologic rationale for DMA is lesions, if unrepaired, can a mutation. Thtre 1s consfderabli In cells "Initiates" the of (4,5); but 1t nay result in of to the (6,7). resulting In may 1n (8,9,10). Protein as can, 1n theory, act is a readily available for DMA. Proportional Ity protein and DMA binding has for a of (11,12,13). Addycts are generally monitored 1n peripheral blood cells rather than tissue, for only § few {e.g., and els platinum) is ind/or lewels ire at {14,151, METHODS Techniques to carclnogen-ONA Indyde ysing adduct-speciflc polyclonal or monoclonal and P-post1abell1n§, Carcinogen-protein nay be antibodies and gas The sensitivity of the DNA to 1s in the of out per 10 -10 nucleotldes. Those it carcinogen-protein quantification also to sensitivity for (16). Identification of particular DNA at low levels Is difficult with analytical cross-reactivity of as the BPPE-I-DNA antibody nhich closely related polycycllc in definitive characteriiatlon of (17). AND Experiment*! Involving chronic to the relationship and macromolecular binding Is genenlly linear with few (12,2,18,3). With to carcinogen-DMA and -protein Investigated in populations with as cigarette PAHs, and nut, iflatoxln and i-nltrosamines, c1s platinyn, psoralen, 4-a«1nobiphenyl, propylene oxldt, vinyl chlor1de°^"nd (3). ------- -49- far the feasibility and sensitivity of the 1n of are by ability 1n the of controls, and illustrate is significant variability In the of and -protein Individuals with or 115,19-25). 1n the involving Is levels of ire 1n so-called controls" (19-20,26-29). of for risk stm 1n the to DMA and 1n and potential in a of Include: identification, of in¥0l¥ed in risk extrapolation and improving the and tlweliness of {19,26,30-321. 1s 1n the following A. Inter!aboratory validation of as ftas recently for (331. B. on the stabllHty of 1n C. of Inter-Individual In levels. 0. on the of In cells and tissues. E. of levels 1n QUA as well as 1n for a of different of F, of critical or "hot on DMA to the of G. of DMA and formation (e.g., and with and chronic to the compound!s)}, H. and on the relationship formation, mutation, and activation. 1. Longitudinal (experimental and on the relationship levels and risk. would be molecylar epidemiologies! In populitlons (sych as to high §nd who experience a high of secondary cancer, or htawily-exposed ------- -50- groups). Biologic could be drawn at the outset and for future analysis, J. to as archives of blood, urine, and tissye for retrospective analysis. ------- -51- KEFERENCES 1. F. 1987, Epidemiology: A New Tool In Prevention, J Cancer Inst, 78, 2. GN, NJ. 1985. and of to Chew-Heals, Environ 62, 5-18. 3. F. The Significance of DMA and in Bi§monitQrin§ Studies. Hut Res (1n pressj. 4. IB, Gattonl-CelH S, iClrschiteler P, M. W, J, Jeffrey A. Cardnogenesls Involves Hyltiple and Multiple cells 1. The Laboratory, Nw York, pp. i. Harris CC» Future Directions In the Use of DMA as for to and Carcinogens. Environ §2, 185-191. 6, Hennings H, R, ML, EF, R, SH Malignant Conversion of Skin Is by Tynor Initiators and by (London), 304, 67-69. 7. E. 1i84. 1n 738, 219-236. 8. FA, Kidlubar FF. 1985. Forntitlon and of Arylaiini DNA In Vivo, Environ 62 , i. CO, KH, Phillips DH, Activation of c-Ha-ras-1 Proto by In _Vfjro Modification with the Cheiical Carcinogen, 01 o*PfipoxTle , {London}, 310, ID. K, R, R, K. DMA in J. Res CUn , 112,181-188. 11. L, E, Osteman-Golkirt G. 1983, of tenotoxlc and Relationships of Their Effects. Res 123, 12. HG. and Relationships, 1n: Berlin A, M, K, Ysalnio H (Eds.), Monitoring to and -I ARC Sci, Pybl No 59, Ly§n» pp. 115-128. 13. HG. of as a for Alitylatlng md Arylitlnf Toxlcol, 56, 1-6. ------- -52- 14. SJ, MW. and of S2P 15. E, SH. Zwelllng LA, RF, Polrter MP. Quantitatlon of II (ds platln) -DNA-tntrastrand 1n Testlcylar and J Pin 77, 16. SR, PL. Biological to the Evalyitlon of Risk; of 1n lexical, 4, S367-S370. 17. RM. of New for of to Population Monitoring. its (1n press). 18. Polrler MC» FA, 1987, of 1n and Exp 31, 1-10. 19. F, Rt HK, AR, M» 0, H, Van J, DMA and In and J Inst. 20. f, K, TL, D, Kelly G, RM. 1987b, of Polycycllc In Cells of 21. K, RR, CC. of 1n Cells. 45, 22. A, G, C, K» Trfvtrs GE» HJ, Harris. CC. of Polycycllc 1n the Urine, in and to the in to of Polycycllc In the Rts 46, 4178-4183. 23- MS, PL, SR, M. 1987. of in and Cancer Res 47, 24. IP, HF. 32p Of DMA 1n Oral Cells. 7» 111-5-1120. 25. Phillips DH, A, PI. DNA 1n Human and Perlphtral Blood Carcinogenests 7, 2071-2075. 26. BA. 1980. An to the of the Risk to Han DNA Toxicol, 3:271-281. ------- -53- 27. Wright AS. 1983. Molecylar Dosimetry Techniques 1n Risk An Industrial Perspective, in: AW, Schnell RC, Miya TS {Eds.). in the Science and Practice of Toxicology. Elsevier, pp. 311-318, 28. Tornqvist M, Os Golkar S, Kaytiainen A, Jensen A, Farmer PB» Ehrenberg L. 1986, Ttssye of Ethylene Oxide in Cigarette Determined Adduct Levels in Hemoglobin. Carcinogenesis, 7, 1519-1521. 29. RB, RM, Santella RM, Cefalo RC, Avltts TA, R 1986. Detection of Smolcing Related Covalent ONA Adducts in Placenta, Science 231, 54-57, 30, BA, BE, IB. Banbury 1982. Indicators of Genotoxic Exposyre; Report No. 13. Cold Spring Harbor . Lab, Cold Spring Harbor, NY. 31. MAS Briefing Panel. 1983. Report on Effects of Hazardous Chemical Exposures. National Sci, Washington, DC. 32. Sobsel FH, 1982. The Parallelogram; An Indirect for the of Genetic Risks from Chemical Mutagens. In; Progress in Mutation Research (Bora KC» Douglas GR-, ER. eds.). Elsevier, pp. 323-327, 33, Santella Rm» A, F, et al. 1987, Intelaboratory Comparison on Antibodies and Immunoassays for Benzo[a]pyrene Diol Epoxide-i Modified DMA. (Submitted). ------- -54- Ep1demiolog1cal 1n long-term to can which, on the and of can of §nd to and personality to and irreversible to be an early of neurotoxldty. led the international to call for for Identifying and 1n and 1n clinical OF AND An of to applications to this 1s the of §nd The of this field 1s to how function is out by the as well as how various neuropithological conditions, is Alihelier's and Korsakoff's cognitive 1n of Is By fully i of all in the basic is with the neyroblology of leirnlng ind It 1s not surprising 1n this 1s occurring it a This will briefly highlight specific in this field i on neyrotoxicological Analysis of the neyrob1o1o§y of leirnlng his (1) key brain I.e., brain are essinttil for different of (2) "circuits" in the brain, I.e., the of information results 1n the of and {3} 1.t., the of the during learning, and the and cellulir which under!1ne The first of has hid, as one of Its concerns, the of how to from animal of cognitive dysfunction to The latter two primarily with analyzing the at molecular levels, In the 5-7 years, discoveries in ill of in the of extrapolation two forms. One has to develop behavioral in animals which are to which are to cognitive function 1n The form, and the one which we will here, has to apply ------- -55- behavloral to which are to which ire will behivforally and neuroblologically, in For It has recently delafed-non-mitching-to-saiiple, i which is i indicator of with limbic and frontal cortical in and prinates, 1s also a sensitive indicator of with similar in . clinical is the use of conditioning to learning deficits, and Alzheimer's which by of eyeblink conditioning 1n rabbits. 1n establish a link the analysis of cognitive 1n and its to research, at validating the appH of new to the of risk will substantially "on the of how can be to risk to populations, following to solvents and pollutants. The important the sophistication of neyrotoxlcological 1s the Identification of neural circylts sybserving learning. The of this 1s the neurobiologlcal of conditioning. This Pawlovlan conditioning has for neurotoxlcologlcal including: (a) the of of Its properties, which It to associative to In a single experimental preparation; (b) the ability to directly qyantltative of and behawlor, on-line and in real time; (c) the ability to directly the of conditioning In §nfrails'and (d) the of electrophysiologlcil loci for, In the brain EEG activity scilp Important offered by this 1s the of we now Its essential circuitry in the brain and ye also a the effect of on this of and this freatly oyr ability to the of neurotoxicologlcil If an a behavioral effect, we i of to look for Its and effects, and ylt1»ate1y its s) of action. Conversly, 1f a an effect on a or we functional to for in of the of or cognitive which be impaired. Investigators to use Pavlovian of this kind as in the neurotoxicologlcal just this (1987), the has applied to the study of with aluminum toxicity. One final which is mentioning is the use of the i_n vitro brain slice technique to study neural plasticity, ITicTrophysiological studies of hlppocampal slices i long potentlation (LTP), has very influential as an for studying the synaptic ------- -56- of learning. In LTP 1s, In effect, in In efficacy *1th of the cellular ind of LTP a of i (the or of the nay the of LTP, ind may disrupt cognitive function 1n rats. 1s of may be of leg., chemicals). It is likely in this may be as a of for their ability to and of characterizing the of any are foynd. SUMMARY In of 1n i for the analysis of 1s at a Importantly, a level. It 1$ possible to «i scientific subdlsciplines, including cell biology, , neyrophyslology, and ind 1n i and way to (a) identify the ri»k to n§rm¥Fc¥pTt1 ve function or (b) the of of as the wKTcF^reTiown to loss, and ------- -57- USE OF IN of which his for application to a variety of 2 for others). This will new applications in neyrotoxlcity. to a foreign elicits an by production of are proteins that with are of inducing antibody formation). anttgenic can viruses, bacteria, proteins, or like envlronnental chemicals. Ant1gen-antibody reactions ire highly specific, the specific to biology. It 1s this specificity of the intigen/antibody that has exploited by the scientist with applications ranging curing Polio to understanding the molecular of catalysis. Antibodies are in the by B lymphocytes (B-cells), each of which its own antibody. In theory, as as 10 million antibodies cm bt by a 1n to a single antigen. with a an an^ contains one B-cell can only one but are B-cells epitope, this fs to as a polyclonal (many cells) antibody. The fusion of Kohler and Milstdn, for which the Prize, was to the 1 Imitations with the use of polyclonal antibodies (e.g., con nation, heterogeneity, limited supply). The antibodies by Kohler and Mil stein referred to as monoclonal by a single {mono} B-cell line {clone}, antibodies including: 1) speclfldy clone only one specific antibody}; 2) unlimited sypply largi of antibody and can be Indefinitely); and 31 purified antigens are not for the production of antibodies by definition only a single antlgenlc determinant). ftonoclonals to define, localize, purify, quantify, and modify antigens. The distinction the yse of monoclonils, is to polyclonal antibodies, is confer fir precision and accuracy and are available as essentially , off the shelf reagents. Thus, It 1s now possible to define with § greater of certainty This tnit of monoclonals has it far to identify rare in vivo and ir» vitro (e.g., tissue cell and typing 1n celTTiTltureTT """One" example of the application of monoclonals that is relevant to EPA is the usi of specific monoclonals to identify dioxin In contaminated soils. Trye pyrficition of antigens from (e.g., strum, tissue) also is now possible with aonoclonals. Thus, rare factors or hormones, as interferon, can now be eisily in ------- -58- Likewise, of in H to polyclonil an chorlonfc for 8y specific modification of toxtdty or also be realized. are of dlfoxln (with antibody to dlgoxin), and with linked to to cell antigens. of Monoconal sto The of on nonoclonal followed and Milstein's original in 1975 are now beginning to revolutionize ntyrobioloay by providing the tools with to the cellulir and subcellular organization of the Thus, the Impact of antibody technology on has the identification of cell 1n the monoclonal s now which identify previously of and g11i (the cell of tissue! which would not to be different yslng classical of light or electron microscopy. Monoclonals also crucial for the Identification and of and to differences within the nolecule. For example, monoclonal antibodies that reveal phosphate-containing nonphosphate-contalnlng neuro filaments, the major strycturil {filament) of ill neurons, The significance of this i.e., the or of a single 1s this may be to i ¥ariety of neurological Including Alzheimer's and also may i to Injury of the In neyrotoxlcology » It 1s toxicant-induced Injyry to the or 1s by alterations In the of specific regions. Furthermore, within an region, the to injury nay cell distinguish the cell comprising the by monoclonal antibodies, cm be to detect, localize and ce'-jlar to This can be by a as are as specific for microscopically localizing specific within animals. Quantitative are with the antibodies by using monoclonal-bised Thus, through the yse of monoclonal in /blochenlcal cvalyatlon of neurotoxicity may eventyally be The possibility exists the sensitivity and specificity of monoclonal antibodies can be applied to the and of into the cerebrospinal fluid and blood is a of neurotonic Theoretically , it would possible to 1nexpert$l¥e monoclonal -anti body for neurotoxlcity In the population. In summary, it is clear that cyrrent in neurosciences will continue to re¥ea1 the extensive cellylir and subcellular heterogeneity of the on the yse of monoclonal antibodies. The EPA, by ------- -59- in will by the to mS ------- -60- A problem In environmental health 1s the non-Invasive of small or at early applications of hold for just In the few since liuterbyr1$ fij was published, (MR) his rapidly into an clinical and, also, a of potential* with high field, (1.5 to 4,7 Tesla) are available ind ire for and laboratory animals the of density, relaxation tines, and the acquisition of 3-dlraensfonal types, of and allow the of or perfyslon (2-5). for of and Intensity, the (proton) Is for the production of.orictically all MR Images, The abilities,to nyclei (e.g. Ma] and chemically shifted nude! (e.g. H In fat} ind the versatility and of the Present day MR of and laboratory animals detail that, in applications {biologic and small animals), microscopic levels. In publications (6,7), of frog and plant with {voxels) of 0.2 and 12.0 L, respectively. are it Un1«r$ity In chemically as as 100 I in 1n fits. The ability to in live Imaging (is long as 6 hours!, fields and gradients, pylse and little or no relttlwe motion. respiratory 1s the to the liver, the list {no 1s by the , y$ing a anesthetic, and synchronizing signal acquisition to respiratory (8,9). of the of MR are to of techniques, and are unique. Similar to {CT} scans, MR imaging 1s non-Invasive ind may be multiple on the anlia! or patient. In toxicology experiments, for example, Incorporation of MR Imaging of a of animals could provide important Information concerning organs* to lesion {e.g., tumor) and to or modified (e.g., or of lesions). MR animals per with conventional for similar information. While Imaging on Ionizing radiation are well established, rapidly (a distinct to MR Imaging ------- -61- it its of development), and excellent for demonstrating structures or (e.g., lesions containing calcium deposits, MR has distinct and Important advantages. With eurrtnt and anticipated fields, gradients, and RF signals, and with the proper precautions MR imaging 1s considered for patients and technicians (10). Additionally, the MR signil, unlike the penetrating of Ionizing radiation, contains Information 1n addition to of tissue (in this case, density. The signal 1s by the at which relax 1n relationship,to the nolecylar lattice (Tl, spin-lattice, longitudinal relaxation! and to other (T2, spin-spin, transverse relaxation). are Influenced by the composition of the tissue {probably by the and motional of molecults}, the resyltlng can permit distinction of are similar In density but differ In relaxation Although not a consistent finding, malignant frequently Tl and 12 relaxation greater of benign or ndmil tissue, concerning the Inability of MR Imaging (relaxation to distinguish pathologic entitles (111. This nay be partially related to the acqyisltlon of the signal from tissue si lets that, of slice thickness, Include and normal within and adjacent to the lesion of Interest. In animal it University, this possibility is being explored by excising very thin (only 1,26 mn thick) tissue slices 1n rats. While signals thin slices art and Imaging sessions are relatively long, the thin sections with high resolution greatly Improve the selectivity, and, hopefully,, the discriminating ability of the AND In clinical medicine, MR Imaging cowpllmtnts and frequently the performance of other Imaging methods. MR imaging excel!s In demonstrating ntoplastlc, dentyellnating, and degenerative of the central system, of the sy$cetib1Hty of the thyroid md parathyroid glands to Ionizing radiation, MR imaging is a preferred for nation of tissues. Respiratory and cardiac gating to prodyce excellent diagnostic of the heart,, thoracic blood vessels, and lungs. MR of liver, kidney, reproductive organs, and pelvis routinely a variety of neoplastlc and non-neap!astic processes. Current and futyre will incorporate the use of faster scanning sequences, 3-dlraenslonil Imaging, of perfyslon and flow, contrast Imaging with jjn vitro spectroscopy of different nyclei (e.g., 1PS C, Nas FJ» chem1caT~sh1 ft Imaging (e.g., permitting proton of H. 1n fat), and, possibly, myltinuclear Imaging (e.g., P, N). These developments, in addition to Improving the sensitivity of detecting lesions, will allow Imaging to be with j_n vivo metabolic studits can characterize biochenical activities 1n a "FegToi~of interest. In toxicologic experiments, techntqyes that allow prolonged anesthetization of rats (as long is 6 hours) associated with respiratory and cardiac synchronization for thoracic and abdominal ------- -62- imaginf. High field (300 MHz, 7 Tesla) are being developed and that a theoretical resolution of 10 M. of active include the of RF coil designs, and the use of stronger field gradients, surface and implanted coils, and contrast agents. Within a few years, increases in resolution should permit, for example, the visualization of renal glomeryli, preneoplastic hepatocellular foci, and nyclei in the br§in. With developaents, Lauttrbur's closing in his 1973 paper would remarkably prophetic, "Zeugmatographic techniques should find useful applications in studies of the internal structures, states, and compositions of microscopic objects," ------- -63- REFERENCES I. Layterbuf PC. Formation by Local Interactions: Employing Nuclear Resonance, 1973; 242:190-1, 2. CJ» WR. The Evolution of Nuclear Magnetic Resonance. In: Introduction to Magnetic Resonance Imaging, Denver; Multi-Media Publishing, Inc. 1984:1-12. 3. Andrew ER. A Historical Review of NMR and Its Clinical Applications. Br Ned Rev 1984;40;115-9, 4. Damadian R. Tumor Detection by Nuclear Magnetic Resonance, Science 5. Lauterbur PC. Cancer Detection by Nuclear Magnetic Resonance Zeygmatographic Imaging. Cancer 1986 ;57: 1899-1904. 6. Aguayo JB, Blackband SJ, Schoeniger J, Mattingly MA, Hinterraann M, Nuclear Magnetic Imaging of a Single Cell. Nature 1986 ;322: 190-1. ?, Johnson GA, Brown J. , .Kramer PJ. Magnetic Resonance Microscopy of Changes in Content in of Transpiring Plants. Proc Acad Sci USA 1987 ;84:2752-5. 8. Hedlynd L, Oietz J, R, Herfkens R, et al . A Ventilator for Magnetic Imaging, Invest Radio! 1986;21:18-23. 9. Hedlund L, Johnson GA, Mills GI. Magnetic Microscopy of the Rat Thorax and Invest Radio! 1986;21:843-6. 10. Saunders RD» Smith H. Safety of NMR Clinical Imaging. Br Med Bull 1984;40;i48-54. 11. Johnston DLS Liu P, Wisner GL, Bi, et al . Magnetic Imaging; Present and Future Applications. Can Med Assoc J 1985;132:765-77, ------- -64- Luster In a broad irmunotoxlcology can be as the study of (inadvertent) effects of envlronnental chemicals, therapeutics or biologicals on the The of effects that nay occur include {i.e., or hypersensltlvlty (allergy) in Instances, autolnmunlty. A large of Information has over the 10 years that to certain chemicals or can dysfunction and alter resistance in animals following and sybchronic exposyre. Examples of are listed 1n the attached table, The extensively of environmental Is the aromatic .(PHAs), Including polychlorlnated biphenyls, polybromlnited biphenyls, chlorinated dlbeniofurans §nd the of this class, chlorinated d1benzo-p-d1ox1ns. the variability the toxic of 1n laboratory animals during or adylt life with and, 1n pirtlcylar, dibenzo-p-dloxins Indicated that the 1s one of the sensitive for toxicity. These effects are characterized by thymlc and and persistent of cell-media ted (T. cell) and featyres of neonatal Laboratory further Indicated the cell for by 1s the epithelium which 1s for T cell only a United of reports Indicate dysfynctlon following to PHAs, the effects to be remirtedly similar to which in animals. For example, sypprisslon of a hypersertsltiv1 ty and susceptibility to respiratory Infections 1n who accidentally polychlorinated blphenyl/dibinzofyran-continlned rice §11. of this deregulation by PHAs his 1n Michigan residents inadvertently polybromlnated blphenyls. ThiSi Individuals also persistent of cell-mediated Irani unity with of null cells, reflecting the of Gills. Althoygh long-term of polybrominated biphenyls re«i1n to be determined in hynans, early data Indicate a correlation alterations and incidence, Thus, 1t early laboratory In a of the Iranunological dysfynctlon is in following Inadvertent exposyre to these compounds. ------- -65- or 11 MB Class v (Mr Laboratory * PCB + FB0 + 1CT + * * + + + * BftP * + * * * + + T: : SQ2 + ;;" ' + t M.S. . M.S M.S. M.S. M.S, U.S. N.S, N.S. N.S. + N.S. 4- M.S. U.S. «„!. • + ------- -66- and WUcox BACKGROUND Public scientists long the of pollution. One ind troubling has are of on the earHtst of pregnancy. This can Include environmentally- early abortions/fetal If way to the earliest of pregnancy, of , , or drug be easily and It is 151 of clinically-recognized end in loss {spontaneous abortion). The risk of loss has to b6 in populations with occupational. environmental, etc,, clinical don't tell the story; clinically-recognized only a portion of all losses. There are at as earlier as Thus, a which could pregnancy very eirly define its ending precisely could help pinpoint chemical or other environmtntal exposyres night Involved In ending. The application of new with chorlonlc offers possibilities. METHOD Detemination of early loss reqyfres sensitive and specific for identifying The of to one of the subunlt of HCG has vastly the of HCG to early pregnancy, HCG 1s by the starting at the diy fertilization. HCG 1s quickly in the mother's urine and is by 1mmunometr1e For thH reason, HCG are the of studies of early Tni3 1s to the of the HCG molecyle. The is up to one sensitive than any previoysly available This sensitivity has to be Important up to of early losses a 1e¥el of HCG secretion could by IMPLICATIONS Eifly loss way be one of the earliest signs of to or toxins that reproduction. It should be to streamline this of study, collecting yrlnes only on early loss is likely to be This could be to high-risk groups of 1n occupational or other settings toxic effects on reproduction are suspected. are now able to HCG in yrint to the levels occyr in non-pregnant are just now beginning to be ------- -67- 1n for the of loss. This 1s in exciting new applied in Is the of 1n biology. This may be a for and and clinical are 1f we ire to 1n vulnerability to exposure. Further and is in this -- as a high priority -- of are can the of clinical If may Inclydi of the and the microelectronics Industry. ------- -68- 8 OF Hoel/Mlchiel AND RISK Sinct relevant ep1deffl1o1o§1c and clinical Information are lacking on the potential health with to a specified or chemical, laboratory animal usually the for qualitative and risk The majority of inlmal-based, Is qualitative 1n nature. That 1$, laboratory or Identification of a as a potential 1s sufficient, 1n and of itself, to control or future of the public to the or in question, and no of the of the risk involved In the may be (*.g., regulation of potentially the 1t 1s the of In the qyant1ficat1on of health risks that 1s of scientific and Art1nal-ba$ed, two or be extrapolation, by the high levels typically 1n laboratory animal and, of the ultimate Is with the risk to the single 1n 1s the of the specific or to be 1n determining the risk or the level for the consideration. In cardnogenes1sf may as far ts 1s or further Insights into the underlying the carcinogenic Certainly the for on the meaningfyl Incorporation of molecular and risk, 1s well and It offers in opportunity to in the quantlflcation of potential risk on For noncarcinogenlc or Is definitely i to the "safety factor" to which has the the mid-501s, 1n Instances, to the of quantitative similar to in ctrci nogenesis. Regardless of the toxicologic of Interest, howe¥tr, it 1s clear that, increasingly, will be on the or extrapolation closely reflect the underlying biological mechanisms. For example, 1n cardnogenesls the question of "primary" "secondary" or "indirect" of action and their on the risk 1s with who the litter traditional low-dose extrapolation (1), On the who out of ------- -69- or on the for levels for may to the biological ynderlie its giving particyTat to the of thresholds* For 1f one a threshold is the i (biological),"no effect" or a or level the are minimal"? It to the population as a or Individual to Individual? (In the latter the nay be one for which no threshold That 1s, 1f 1 evils vary indlvidya1s9 the "population" level to the for the stnsittve 1ndi¥ldyi1 tn population, which, for all be indistinguishable t level.! of 1s 1s a (is to traditional) for the of any to be an In of levels (2). The of may well as scientific as the of the Certainly, the utility of the for Identifying risks 1s broadly within the scientific [e.g., see the (3) the interpretation of to carcinogenic risk or clinical are not available]. Is no unlversilly of the results 1n to 1s usually is to and risk is 1n of the Yet, e.g., on can by as as 40-fold (4) on are in of an daily or a ng (divided] by weight. Furthermore, one to rely on a as the for extrapolating risk an 1s only an for the variety of can to In (e.g., In llfespan, size, profile^ genetic etc.). In the quantitative of taxied ogle will on the use of nolecylar and For the use of or nolecylar dosinetry, scientifically feasible, to the "biologically could significantly the extrapolation of toxlcologlc for quantitative risk estimation and are Increasingly to epidemic!ogle as well as to laboratory animal resylts, particularly In the of cardnogenesls. of the Inclyde fitting of the to DoTTs 16}, Day and Brown's y$e of the to a of risk as ------- -70- and radiition early, or early and late of the 17]» Ill's (6) use of and to the of site-specific syrvivors, and their use of linear, linear-quadratic and to predict risk with Ionizing radiation. While the use of obviously elimlnatfs the for extrapolation, nay not be sufficiently sens1tl¥t to allow one to or, in instances, to if any risk to be low or levels of A of may be to increase the sensitivity of the available epidemlologlc For Instance, Initial it risk Identification and could be on sens1tl¥e within the population as the old or young, indivldyals with insufficient Individuals suffering concurrent or Inherited deficiencies, and individuals also to risk factors for the toxlcologlc endpoint or effect of Interest. Recently, a new speciality his In the field of epidemiology, which 1$ as nolecylir or biochemical epidemiology. One of the primary of molecylar 1s to laboratory for the identification and characterization of to epidcmlologlc field studies, so as to clarify the of underlying relationships, I.e., relationships and or toxicploglc (8). Specifically, may provide quantitative of (e.g., blood levels}» specific (e.g., ONA formation), biologic and early or frank to replace the subjective and qyalitative 1n ep1dem1olog1c Investigations (e.g., histories and classifying as tlther "exposed" or "unexposed".) While Interest 1n ind and application of biochemical 1s increasing rapidly, validation of their use for epidemiology 1s currently a major and 1t 1s likely to to be so in the future. [Among the be In any validation exercise the determination of marker sensitivity, specificity, predict!¥ity, of nomil or baseline values, whether marker 1s reflecting cyrrcnt or cumulative exposyres, or and cumulative or noncymulative biological (8),] POPULATIOM The last in the qyantitatl¥e risk 1s the of the overall risk for the popylatton of or, alternatively, the of an level for population. of the uncertainties Involved 1n using experimental animal or epidemic!ogle 1n Identification particularly, 1n dose-response modeling and low-dose risk estimation enumerated. If a can be for the of a ------- -71- tnreshold and a factor approach Is elected, 1t 1s Important to that failure to adequately for the ynknown, underlying threshold can result In a proportion of the population having their Individual threshold values falling the estimated level In Instincts (2). Another significant factor that must be 1n developing population risk is the or estimation of levels within the population yrtder evaluation. There art a of potential or yncertalntlis typically Involved 1n the estimation of population exposyri levels. Exposures vary considerably Individuals or for a single Individual across time, so that the use of exposure levels may not be very representative of the histories of Individual population While use of worst-cast exposures providt in on the actual levels of exposure encountered, 1t can also lead to an overestimate of the popylatlon's health risks and certainly a of uncertainty estimates. The uncertainty 1s or worst-cast estimates are multiplied by the risk per unit to obtain an overall estimate of population risk. For example, though worst-case may the experience of much or possibly all of the popylatton of Interest, "iveragf" risk per unit estimates may significantly the risks of the susceptible of population. the uncertainties Involved In quantitative risk estimation and concern for the health of the popylatlon often led to the overuse of worst-case or upperboynd 1n quantitative risk est1mat1on--assumpt1ons that result 1n is undyly conser¥itive estimates of the popylatlon risks. However, are other investigators (9) who national the of health risks has to be alnostly exclusively on cancer risk, and as a result, other (perhips less quantifiable) of human disease or dysfunction may have received Insyfficient attention: (See Appendix), If this 1s the cise, then, 1n any specific situation the estimated "acceptable", "virtually safe" or "minimal risk" for cardnogenesls may still entail an unreasonable le«l of risk of health the estimation has on conservative assytnptlons. The OSTP (10) tnd other science policy reports the for qualitative and quant1tatl¥e characterization of the uncertainties of specific risk (e.g., consideration of the Impact of model selection, the use of one set of laboratory over another, the choice of a particular species as being most representative of humans, etc.). Also Important are considerations and specification of the assumptions ynderlylng a particular risk assessment (e.g., the construct of an estimated lifetime dally so animals continuously at a constant throyghoyt their lifetimes might be to estimate the risk in hunans who miy received Intermittent at varying for only a portion of their Hfespan). The continued attention to/stress on descriptions of specific uncertainties and assumptions involved 1n any given risk assessment and to their potential impact on the estimation of risks has helpful to ------- -72- with regulatory responsibilities for rational and decisions the fate of the consideration. ------- -73- REFEREI4CES 3. 4. 5. 8. 9. Hoel , D.G., Baseman, O.K., Hogin, M.D., Huff, J., and McConnell , E.E. The Impact of Toxicity on Carcinogenicity Studies: Implications for Risk Assessment. (Sybmitted for Publication) Portier, C., and Hogan, M, (1987). An Evaluation of the Safety Factor Approach in Risk Assessment. In: Mclachlan, J.A. , Pratt, R.M. , and Markert, C.I. eds, Banbury Report 26; Developmental Toxicology: Mechanisms and Risk. Cold Spring Harbor Laboratory, New York, International Agency for Research on Cancer (1985). Preamble {p, ZO). In; Volume 35: Polynuclear Aromatic Compounds, Part 4, Bitumens, Coal-tars and Derived Products, Shale-oils and Soots, IARC, lyon, France. Office of Technology Assessment 11981), Assessment of Technology for Determining Cancer Risks from the Environment. Washington, D.C.; Government Printing Office. Doll, R., and Peto, R. (1978), Cigarette Smoking and Bronchial Carcinoma: Dose and Time Relationships Regular Smokers and Life-Long Non-Smokers, J. Epid, Conro. Health 32: 303-313, Day, N.E., and Brown, C, C. (1980), Multistage Models and Primary Prevention of Cancer. JNCI 64: 977-989. National Academy of Sciences, Committee on the Biological Effects of Ionizing Radiations (1980). The Effects of Populations of Exposure to Low Levels of Ionizing Radiation; 1980, Washington, D. C.: National Academy Press. Schulte, P.A. (1987). Hethodologic Issues in the Use of Biologic Markers in Epidemiologic Research, Ant. J, Epid. 126; 1006-1016. Silbergeld, 1399. E.K. (1987). Letters: Risk Assessment, Science 237 10, U. S, Interagency Staff Group on Carcinogens (1986), Cl nogens: A Review of the Science and Its Associted inlPt. FHP fi7 • ?nU7ft?_ Circinogens; Principles, 1986), Chemical EHP 67: 201-282. ------- -74- Of AND Chernoff and The of basic on cancer and non-cancer within any orginizitlon 1s a variety of as Congressional the given organization"s operational policy, public perceptions and concern, ongoing Identification of potential §nd to extent, the of disciplines by the organization's scientific staff. All of an 1mp°act on scientific in their designing and a basic to their organization's now and In the future. Generally, 1n legislative authority EPA to evalyiti a of potential health effects associated with to environmental chemicals ind Insults, Rartly legislation specific health to be irtdpolnts. It 1s, therefore, EPA's policy which dlrtcts to specific of for environmental a public Institution, EPA Is influenced by the and concerns of the public and Industrial of chemicals, Consequently, EPA Its administratfve and regulatory policy to For years the primary envlroontntal health concern, as by the public, the possibility of chemically-Induced cancer. The for this concern Include the and Irreversibllity of the Its for debilitation and lethality, and the chaiicals to which is prevalent cm in laboratory animals. As a result of concerns the body of that has years, EPA's (as well as regulatory regulatory policy has been largely driven by as the htalth endpolnt of greatest severity. the however, it has Increasingly art other health endpolnts which nay be and by to environmental agents. The methyl mercury- of birth in Japan, the incidents of delayed neuropathy in the Middle East, and the occurrence of sterility in occupattonally to chemicals 1n the USA all to alert the public the potential risk of to may require consideration of health endpolnts. The of this realization has a broadening of the of concern and a simultaneous of to additional Along with this has an increasing to consider during the formulation of regulatory policy. Tox1co1og1sts in the public and private also identified other and susceptible populations that are at potential risk from to environmental These realizations lead to considerable for research 1n as ------- -75- immunotoxlcology, herltible ind and geriatric Additionally, and the public increasingly the of lifetime to relatively of § of to i of the of is as the to risk. Finally, are two of the and the scientific of The translation of Into risk 1s an difficult While a is desirable, the regulation md of faulty letd to finclydlng a in the of life of and is the Indicates, now and in the future will be to the scientific and of risk The of a of the for and non-cancer 1s yltimttely the rational «ay in which to foraylate policy. This to i continuing for The allocations the for simpler, and of testing. Thi of and of 1s fir in of ability to for by by the the use of animals in i to the of alternative The EPA efforts 1n greatly the last for the listed has led to i and non-cancer 1s impossible to since art so factors go into the of this Ctrtalnly, § allocation to and h« the to utilize a of health in the fenny lit! on of regulatory policy. Long-term Into both and 1s recognized is 1f the Is to a policy In the possible, consider and in the will multiple tox1co1og1ci1 the of and se¥er1ty of effects will be attention. Efforts will be to ind lexicological in a fashion. win the of structure-activity and modelling, design, §nd of as well as endpoints. ------- -76- 6 This century has the of abnormalities 1n early growth and development, chronic and cancer as the major of morbidity and mortality 1n the Industrialized nations of the world. Initially, often as the result of heredity or the natural of the aging process. recently, has a rtcognttion frequently important or risk factors 1n their etiology. of environmental risk are either directly by or subject to their manipulation. They include chemical and physical in the air, water, supply, drygs, products, and workplace. While of the of risk are difficult to or verify, 1t his ¥arioy$ly i significant of the two million Individuals who die year in the United may had their lives to by the effects of air pollution; that pollutants In oyr drinking water nay play a role 1n the of and disease, which are the two leading of 1n this country; and the collectl¥e of and raiy now be a level of typically with Therefore. and regulators are Increasingly to Identify risk factors and §r eliminate their effects, Attention has on of the cin be one to the Impact of environmental on health. Often, relevant epldwrfologlc and clinical Information on the potential health with to § specific or will not be available. is available, 1t may not be sufficiently sensitive or specific to allow an to that to the unknown, underlying relationship and In the available may not permit one to 11 any health risk to be with low or Itvels of As a result, laboratory anintal will constitute the primary basis for both qualitative (I.e., Identification) and quantitative risk estination, of the high {often maximally tolerated) typically 1n laboratory quantitative risk estimation on Involves two be low-dose extripolatlon and extrapolation. In instances (e.g., when the of Is i carcinogen or mutagen) mathematical modeling will be to low-dose risk estimates, and choice of a particular nay a significant impact on the magnitude of the risk. In a threshold may ------- -77- be ind a to levels. This clearly a of and Ideally, the of be by Into all of the species-specific coyld contribute to differences in to the of the to Is to and the In mm are on in scale. Unfortunately, of the be justified on biological and significant differences can resylt In the on the As our on risk to and 1,s to the to the uncertainties with risk and to uncertainties by luprovlng the biological which the risk 1s In to to the Itself,'Which ind on §yr efforts to the risks by are I of our to the of environmentally-related are the of on the majority of the the Insufficient and training of our nation's physicians with to and the surveillance of populations or potentially to Priority be to 1n a of 1f we ire to and oyr of the role of en¥i factors in For to be to the and of {particularly non-Invasive for low levels of to Similarly, we to a of the biological environmentally-related to the of health ind the primary/secondary of environmentally-related A of of long-term, activities 1n or nay direct application to the of EPA and regulatory on an basis art In this Comparison of of (I.e., cellular during and activation 1n and chemically-induced may Insight Into the of formation at the raoltcylar level, In addition, of the uncertainty invoked 1n of carcinogenic risk eientuilly be by comparisons of activation and expression. ------- -78- Recent advances in biochemistry and molecular biolofy have led to the of highly sensitive which may the quantification of the Internal of carcinogens or 1n the biologically effective in tissues. This ability to external exposure or adwilnl levels on a biologically-relevant basis should e¥cntya11y lead to a clearer understand!ng of the relationships and or toxicologic effect for health in the environment. Recognition of the potential usefulness of biochemical has led to the of a new field of epidemiology, is molecular or biochemical epidemiology, has as one of its major goals the adaptation of laboratory into eplderafologic field studies. In the fields of neyrotoxlcologj and Immunotoxicology new methodologies promise to lexicologists to greatly Improve our ability to central and deficits. The utilization of novel techniques in molecular biology {e.g., monoclonal antibodies to specific critical chemical components of systems) promises to allow .improved evaluations of potential disfunctions. In the of reproduction one of the important questions involves the potential of environmental to affect pre-1mplantat1on loss. Researchers recently Identified an antibody to a sybuntt of the hormone human chorionic gonadotropin. This advanct enables the identification of spontaneous abortions at an earlier and with accyracy was previously possible and may significantly improve oyr monitoring capabilities. In addition to identifying specific of long-term activities that either §re generating or may results directly applicible to the environmental health issues that EPA from a regulatory viewpoint, this also to describe the relationship long-term and short-tern (or Immediate) "problem-solvlng" research to put 1t 1n perspective. For example, It is noted that the general philosophy undtrlying basic health research is that ynderstanding more about the biologic by which environmental hazards as toxic chemicals Induce effects will lead, ultimately, to earlier of effects, sensitive analytical for fully characterizing their potential Impact on health, and a understand!ng of how to eliminate or, at least, that impact. The dlstlngyishing characteristic of this basic research 1s that 1t typically "generic" scientific and 1s not focused on a specific or concern. Fyrtherroore, it usually be for a period of several 1t results may a direct application to regulatory or problems. The environmental health with the toxic lead 1s to illustrate the necessity of and role for long-tem research activities In the of a scientific foundation for constryctive actions dealing with public health While toxicity resulting "h1§h" level has long recognized as an important public health concern, ongoing, long-term basic has only recently given us the ttchnlcal tools to of the subtle yet extremely effects of low-level lead exposure. ------- -79- Is on a or it nay to be for i of any or cm be is new of the on the the AT 1f the is and the of clearly any 1s i of and may be the Is Certainly, 1s the In ind to a with It clear, of the {or EPA an If not the and 1n the only to will be to on the and insights long-ttm This certainly has the in with to The will us to in 1s to stable, consistent for a With and EPA can issyes It 1s particularly and its own findings and of and institutions to the of critical ------- |