1. Hormesis: What it Means for Toxicology, the Environment and Public Health Edward J. Calabrese, Ph.D Environmental Health Sciences School of Public Health University of Massachusetts

2. Overview How I Became Involved with Hormesis Hormesis:Toxicological Foundations Examples of Hormetic Responses Comparison with Threshold Model Hormesis and Risk Assessment

3. Hormesis Definition: Dose response phenomenon characterized by a low dose stimulation and a high dose inhibition. Generally similar quantitative features with respect to amplitude and range of the stimulatory response. May be directly induced or the result of compensatory biological processes following an initial disruption in homeostasis.

4. HORMESIS Interpretation: Issue of beneficial/harmful effects should not be part of the definition of hormesis. This assessment should be reserved for a subsequent evaluation of the biological and ecological context of the response.

5. Response Dose A B (A)The most common form of the hormetic dose-response curve depicting low-dose stimulatory and high-dose inhibitory responses, the  - or inverted U-shaped curve. (B)The hormetic dose-response curve depicting low-dose reduction and high-dose enhancement of adverse effects, the J- or U-shaped curve.

6. Hormesis and Evaluative Criteria Assessing the Dose-Response Continuum: LOAEL-defining the toxic phase of the dose response NOAEL (or BMD)-defining the approximate threshold Below NOAEL (or BMD) doses-number and range Concurrent Control

7. Hormesis and Assessment Criteria Dose Response Patterns Statistical Significance Replication of Findings

8. Evidence of Hormesis General Summary: Hormesis databases: thousands of dose responses indicative of hormesis Hormesis is a very general phenomenon: independent of model, endpoint and agent Frequency of hormesis: far more frequent than threshold model in fair head-to-head comparisons

9. Dose Response Features Stimulation Amplitude: Modest 30-60% Greater Than Control Usually Not More Than 100% Greater Than The Control

10. Stimulatory Range ~75 % - Within 20-Fold of NOAEL ~20% - >20<1000-Fold of NOAEL ~ 1000-Fold of NOAEL

11. Maximum response (averages 130-160% of control) Distance to NOAEL (averages 5-fold) Hormetic Zone (averages 10- to 20-fold) NOAEL Control Dose-response curve depicting the quantitative features of hormesis Increasing Dose

12. Hormetic Mechanisms Many studies have provided mechanistic explanations to account for observed hormesis responses; Each mechanism is unique to the model, tissue, endpoint and agent Some general examples: Often existence of opposing receptors

13. Methanol and Fruit Fly Longevity

14. Gamma Rays and Mouse Lung Adenomas

15. Transforming Growth Factor-Beta and Human Lung Fibroblasts

16. Effects of Acute Ethanol on Overall Social Activity of Adolescent Rats Tested on Postnatal Day 30

17. Effect of X-rays on the Root Length of Carnation Cuttings * * * * * * * *

19. Effect on Growth of Salt Marsh Grass * *

20. Comparative Dose Response Relationships for the Pain Threshold for Vocalization

21. Effect of Different Doses of Morphine on PTZ-induced Seizure Threshold * * * * * * *

22. * Alcohol and Rat Serum Levels * *

23. * MCPA + OAT SHOOT GROWTH * *

24. Effects of Metals on Phagocytosis in the Clam, Mya arenaria, hemocytes

26. Effect of Sodium Arsenate on PHA-treated Bovine Lymphocytes

28. Effect of Gamma Rays on the Life Span of Female House Crickets * * * * * **

29. Effect of Acridine on the Number of Broods per Daphnid ** * * * *

30. Effect of Mistletoe Lectin on Human Tumors in Culture

31. Effects of Ten Estradiol A-ring Metabolites on Endothelial Cells from Human Umbilical Veins

32. Effect of Plumbagin on Human Granulocyte Phagocytosis

33. Effect of Tin (II) on MTT Conversion in C6 Glioma Cells

34. Number of Open Arm Entries in the Elevated Plus Maze in Male C57BL/6 Mice Treated with DHEA * * * * * *

35. The Effects of Allixin on the Survival of Primary Cultured Hippocampal Neurons from Embryonic (E18) Wistar Rats * * * *

36. The Effects of Methyl Mercury on Viability as Measured by Mitochondrial Dehydrogenase Activity in the D407 Cell Line * * * * * *

37. Effects of the Disinfectant Byproduct MX on the Occurrence of DNA Damage in the Comet Assay Using Rat Liver Epithelial Cell Line WB-F344

38. Effects of n-Hexane on DNA Damage in Human Lymphocytes in the Comet Assay

39. Effects of As 2 O 5 on Total Chromosomal Aberrations in Human Leukocytes

40. Effects of X-rays on Chromosomal Aberrations (i.e., Dicentrics) in Human Lymphocytes (pooled results of four donors and six laboratories)

41. Effect of DDT on Liver Foci Formation in Male F344 Rats

42. Bladder Tumor Incidence Adjusted for Time in ED01 Megamouse Study

43. Hormetic or Threshold Which Dose Response Is More Common?

44. The Threshold Model Prediction: Random Bounce Below the Threshold as Practically Defined by the NOA(E)L or BMD

45. The Hormesis Model Predicts that responses to doses in the below toxic threshold zone should be non-randomly distributed The non-randomness should be reflected in the frequency of responses above and below the control value and in the magnitude of the deviation from the control

46. Hypothesis Evaluation Dose-Response Evaluation Criteria Entry Criteria: Estimate a LO(A)EL Estimate a NO(A)EL or BMD One or more doses below NO(A)EL or BMD

47. Testing Threshold Model Predictions Three Separate Database Evaluations: Toxicological Literature - multiple models/endpoints - reviewed 21,000 articles with entry criteria to yield 800 dose responses Yeast Cell Strains - 13 strains/2,200-57,000 dose responses-cell proliferation E. coli – approximately 2,000 chemicals tested over 11 concentrations - cell proliferation

48. Percent Difference From Control Growth Cumulative Percent of Chemicals Mean Prediction Interval 95% Threshold Model Predicted Mean 10 20 30 40 50 60 70 80 90 010 20 3040506070-10 -20 100

49. -20-1001020304050607080 0 10 20 30 40 50 60 70 80 90 100 Cumulative Percent of Chemicals BMD 10.0 BMD 7.5 BMD 5.0 BMD 2.5 Percent Difference From Control Growth

50. Threshold Model Inconsistencies Below threshold responses do not provide evidence of random bounce Non-random responses clearly predominate The non-random responses discredit the Threshold Dose Response Model Findings are consistent with the Hormetic Dose Response Model

51. Why Has Toxicology Missed Hormesis? Modest Response - could be normal variation Emphasis on High Doses - need to define the NOAEL and LOAEL Use of only few doses

52. Why is Hormesis Important? It will change how toxicologists, pharmacologists, risk assessors, and physicians do their jobs It will change the risk communication message

53. Hypothesis Testing Expands Dose Response Spectrum Creates New Categories of Questions

54. Study Design Number of Doses/Concentrations Spacing of Doses/Concentrations Temporal Features –Key feature in recognizing the compensatory nature of the hormetic dose response

55. Implications of New Design Considerations Additional Costs For: Extra Doses Multiple Temporal Evaluations Enhanced Need for Replication

56. Possible Adjustments Less than lifetime studies/different endpoints Less expensive models: cell culture, invertebrates, fish, etc. –increases sample size for statistical power

57. Endpoint Selection Background Incidence: Low Background Disease Incidence Precludes Ability to Detect Possible Hormetic Response

58. Biomathematical Modeling Implications for Cancer Risk Assessment: Models: flexibility to fit observed data; Models: not constrained to always be linearly decreasing at low doses; Low Dose Risk Characterization: include likelihood of below background risks; Uncertainty Characterization: include both upper and lower bounds.

59. Environmental Re-Defining Hazard Assessment Re-Defining Dose Response Default Re-Evaluation of Risk Assessment Practices Harmonization: Cancer and Non- Cancer Cost-Benefit Re-Assessment

60. Therapeutics Cognitive Dysfunction Immune Stimulation Anti-Tumor Anti-Viral Anti-Bacterial Angiogenesis Cytokine/Hospital Infections Hair Growth Molecular Designs

61. Life Style Exercise Alcohol Consumption Stress

62. Perspective #1 The Threshold Dose Response Model fails to make accurate predictions in the below threshold zone

63. Perspective #2 The Threshold Dose Response Model has been significantly out- competed by the Hormetic Dose Response Model in multiple, independent comparisons

64. Perspective #3 There is little toxicological justification for the continued use of the threshold dose response to estimate below threshold responses

65. Perspective #4 Given Perspectives 1-3, there is no basis to use the threshold dose response model in risk assessment practices. This has significant implications for current standards based on the threshold model and future risk assessment practices

66. Perspective #5 HORMESIS: a concept with much supportive experimental evidence that is reproducible

67. Perspective #6 HORMESIS: Based on Perspective # 5 it should be considered as a real concept in the biological sciences

68. Perspective #7 HORMESIS is Generalizable Across Biological Models Across Endpoints Measured Across Chemical Class/Physical Agents

69. Perspective #8 Based on Perspective # 7, HORMESIS is evolutionarily based, with broad potential implications

70. Perspective #9 HORMESIS: very common in toxicological/pharmacological literature, making it a central concept

71. Perspective #10 HORMESIS: a normal component of the traditional dose response, being graphically contiguous with the NO(A)EL

72. Perspective #11 HORMESIS: readily definable quantitative features, that are broadly generalizable, making it reasonably predictable

73. Perspective #12 HORMESIS: far more common than the threshold dose response in fair, head-to-head comparisons; this would make the hormetic model the most dominant in toxicology

74. Perspective #13 The low dose hormetic stimulatory response is a manifestation of biological performance and estimates biological plasticity in the effected systems

75. Perspective #14 HORMESIS: no single specific hormetic mechanism; there appears to be a common biological strategy underlying such phenomena

76. Perspective #15 HORMESIS: important implications for toxicology, risk assessment, risk communication, cost-benefit assessments, clinical medicine, drug development and numerous other areas

77. Perspective #16 HORMESIS: Should Become the Default Model in Risk Assessment – Why? More Common By Far Than Other Models Can Be Validated or Discredited with Testing Generalizable by Biological Model, Endpoint and Chemical Class

78. Perspective #17 HORMESIS: should become the object of formal evaluation by leading advisory bodies such as the National Academy of Sciences