Abstract
Exposure to mixtures of chemicals is an important and relevant environmental issue. Of particular interest is the detection and characterization of departure of biological effects from additivity. Methodology based on the assumption of additivity is used in fitting single-chemicaldata. Interactionsare determined and characterized by making comparisons between the observed and predicted responses at mixtures along a fixed ratio ray of the component substances. Two simultaneous tests are developed for testing for any departure from additivity. Multiple comparisons procedures are used to compare observed responses to that predicted under additivity. A simultaneous confidence band on the predicted responses along the mixture ray is also developed. The methods are illustrated with cytotoxicity data that arise when human epidermal keratinocytes are exposed to a mixture of arsenic, chromium, cadmium, and lead. Synergistic, antagonistic, and additive cytotoxicities were observed at different dose levels of the four-metal mixture.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
ATSDR. (1997), 1997 CERCLA Priority List of Hazardous Substances That Will Be the Subjects of Toxicological Profiles and Support Document, Washington, DC: Agency for Toxic Substances and Disease Registry, U.S. Department of Health and Human Services.
Berenbaum, M. C. (1985), “The Expected Effect of a Combination of Agents: The General Solution,” Journal of Theoretical Biology, 114, 413–431.
Berenbaum, M. C. (1989), “What Is Synergy?,” Pharmacological Reviews, 41, 93–141.
Calabrese, E. J. (1997), “Hormesis Revisited: New Insights Concerning the Biological Effects of Low-Dose Exposures to Toxins,” Environmental Law Reporter, 27, 10526–19532.
Calabrese, E. J., and Baldwin, L. A. (1997a), “The Dose Determines the Stimulation (and Poison): Development of a Chemical Hormesis Database.” International Journal of Toxicology, 16, 545–559.
— (1997b), “A Quantitatively-Based Methodology for the Evaluation of Chemical Hormesis,” Human Ecological Risk Assessment, 3, 545–554.
Choi, E. J., Toscano, D. G., Ryan J. A., Riedel, N., and Toscano, W. A. (1991), “Dioxin Induces Transforming Growth Factor-α in Human Keratinocytes,” Journal of Biological Chemistry, 266, 9591–9597.
Cohen, M. D., Kargacin, C. B., Klein, C. B., and Costa, M. (1993), “Mechanisms of Chronium Carcinogenicity and Toxicity,” Critical Reviews in Toxicology, 23, 255–281.
De Rosa, C. T., Johnson, B. L., Fay, M., Hansen, H., and Mumtaz, M. M. (1996), “Public Health Implications of Hazardous Waste Sites: Findings, Assessment and Research,” Food and Chemical Toxicology, 34, 1131–1138.
Fay, R. M., and Mumtaz, M. M. (1996), “Development of a Priority List of Chemical Mixtures Occurring at 1188 Hazardous Waste Sites, Using the Haz Dat Database,” Food and Chemical Toxicology, 34, 1163–1165.
Gaido, K. W., Maness, S. C., Leonard, L. S., and Greenlee, W. F. (1992), “2,3,7,8-Tetrachlorodibenzo-p-Dioxin-Dependent Regulation of Transforming Growth Factors-α and β2 Expression in a Human Keratinocyte Cell Line Involves Both Transcriptional and Posttranscriptional Control,” Journal of Biological Chemistry, 267, 245591–24595.
Gennings, C., and Carter, W. H., Jr., (1995), “Utilizing Concentration—Response Data From Individual Components to Detect Statistically Significant Departures From Additivity in Chemical Mixtures,” Biometrics, 51, 1264–1277.
Gennings, C., Schwartz, P., Carter, W. H., Jr., and Simmons, J. E. (1997), “Detection of Departures From Additivity in Mixtures of Many Chemicals With a Threshold Model,” Journal of Agricultural, Biological, and Environmental Statistics, 2, 198–211.
Germolec, D. R., Spalding, J., Boorman, G. A., Wilmer, J. L., Yoshida, T., Simeonova, P. P., Bruccoleri, A., Kayama, F., Gaido, K., Tennant, R., Burleson, F., Dong, W., Lang, R. W., and Luster, M. I. (1997), “Arsenic Can Mediate Skin Neoplasia by Chronic Stimulation of Keratinocyte-Derived Growth Factors,” Mutation Research, 386, 209–218.
Germolec, D. R., Yoshida, T., Gaido, K., Wilmer, J. L., Simeonova, P. P., Kayama, F., Burleson, F., Dong, W., Lange, R. W., and Luster, M. I. (1996), “Arsenic Induces Overexpression of Growth Factors in Human Keratinocytes,” Toxicology Applied Pharmacology, 141, 308–318.
Glick, A. B., Lee, M. M., Darwiche, N., Kulkarni, A. B., Karlsson, S., and Yuspa, S. H. (1994), “Targeted Deletion of the TGFβ1 Gene Causes Rapid Progression to Squamous Cell Carcinoma,” Genes Development, 8, 2429–2440.
Glick, A. B., Sporn, M. B., and Yuspa, S. H. (1991), “TGF-β1 and TGF-α in Primary Keratinocytes and Papillomas Expressin v-Ha-ras,” Molecular Carcinogenicity, 4, 210–219.
Hochberg, Y. (1990), “A Sharper Bonferroni Procedure for Multiple Tests of Significance,” Biometrika, 75, 800–802.
Kachinskas, D. J., Qin, Q., Phillips, M. A., and Rice, R. H. (1997), “Arsenic Suppression of Human Keratinocyte Programming,” Mutation Research, 386, 253–261.
Kelly C., and Rice, J. (1990), “Monotone Smoothing With Application to Dose—Response Curves and the Assessment of Synergism,” Biometrics, 46, 1071–1085.
Loewe, S. (1953), “The Problem of Synergism and Antagonism of Combined Drugs,” Arzneimittle Forshung, 3, 285–290.
Loewe, S., and Muischnek, H. (1926), “Uber Kombinationswirkunger. I. Mitteilung: Hiltsmittel der gragstellung. Naunyn-Schmiedebergs,” Archives of Pharmacology, 114, 313–326.
Miller, R. J., Jr. (1981), Simultaneous Statistical Inference (2nd ed.), New York: McGraw Hill.
Mosmann, T. (1983), “Rapid Colorimetric Assay for Cellular Growth and Survivals: Application to Proliferation and Cytotoxicity Assays,” Journal of Immunological Methods, 65, 55–63.
Pietenpol, J. A., Holt, J. T., Stein, R. W., and Moses, H. L. (1990), “Transforming Growth Factor β1-Suppression of c-myc Gene Transcription: Role in Inhibition of Keratinocyte Proliferation,” Proceedings of the National Academy of Sciences, USA, 87, 53–65.
Punnonen, K., Denning, M. F., Rhee, S. G., and Yuspa, S. H. (1994), “Differences in the Regulation of Phosphatidylinositol-Specific Phospholipase C in Normal and Neoplastic Keratinocytes,” Molecular Carcinogenicity, 10, 216–225.
Seber, G. A., and Wild, C. J. (1989), Nonlinear Regression, New York: Wiley.
Stebbing, A. R. D. (1982), “Hormesis—The Stimulation of Growth by Low Levels of Inhibitors.” Science of the Total Environment. 22, 213–234.
— (1997), “A Theory for Growth Hormesis,” BELLE Newsletter, 6, 1–11.
Ye, J., Zhang, X., Young, H. A., Mao, Y., and Shi, X. (1995), “Chromium(V1)-Induced Nuclear Factor-κB Activation in Intact Cells Via Free Radical Reactions,” Carcinogenesis, 16, 2401–2405.
Yen, H.-T., Chiang, L.-C., Wen, K.-H., Chang, S.-F., Tsai, C.-C., Yu, C.-C., Yu, C.-L., and Yu, H.-S. (1996), “Arsenic Induces Interleuk in-8 Expression in Cultured Keratinocytes,” Archives Dermatological Research, 288, 716–717.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Gennings, C., Carter, W.H., Campain, J.A. et al. Statistical analysis of interactive cytotoxicity in human epidermal keratinocytes following exposure to a mixture of four metals. JABES 7, 58–73 (2002). https://doi.org/10.1198/108571102317475062
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1198/108571102317475062