Comparison of the Cancer Risk of Methylene Chloride Predicted from Animal Bioassay Data with the Epidemiologic Evidence

Methylene chloride has been shown to be a lung and liver carcinogen in the mouse; yet, the current epidemiologic data show no adverse health effects associated with chronic exposure to this compound. Hearne et al. have compared the results of a large mortality study on occupational exposure to methylene chloride to the human risk predictions based on the rodent bioassay to point out the inconsistency between the animal toxicologic and human epidemiologic data. The maximum number of lung and liver cancers predicted due to methylene chloride exposure based on the rodent bioassay data was 24 compared to 14 deaths from these cancers actually observed in the Hearne et al. epidemiology study. We assess the minimum risk detectable by the human study in order to calculate the upperbound potency of methylene chloride and compare it to the potency derived from the bioassay data. Results from the epidemiology study imply an upperbound potency of 1.5 x 10(-2) per ppm, compared to 1.4 x 10(-2) per ppm calculated using the most conservative analysis of the animal data. We conclude that the negative epidemiology study of Hearne et al. is not sufficiently powerful to show that the risk is inconsistent with the human risk estimated by modeling the rodent bioassay data. Specifically, the doses to which the workers were exposed, the population studied, and the latency period were not adequate to determine that the risks are outside the bounds of the risk estimates predicted by low-dose modeling of the animal data.     

Experimental Design of Bioassays for Screening and Low Dose Extrapolation

Relatively high doses of chemicals generally are employed in animal bioassays to detect potential carcinogens with relatively small numbers of animals. The problem investigated here is the development of experimental designs which are effective for high to low dose extrapolation for tumor incidence as well as for screening (detecting) carcinogens. Several experimental designs are compared over a wide range of different dose response curves. Linear extrapolation is used below the experimental data range to establish an upper bound on carcinogenic risk at low doses. The goal is to find experimental designs which minimize the upper bound on low dose risk estimates (i.e., maximize the allowable dose for a given level of risk). The maximum tolerated dose (MTD) is employed for screening purposes. Among the designs investigated, experiments with doses at the MTD, 1/2 MTD, 1/4 MTD, and controls generally provide relatively good data for low dose extrapolation with relatively good power for detecting carcinogens. For this design, equal numbers of animals per dose level perform as well as unequal allocations.     

Bayesian incidence analysis of animal tumorigenicity data

Statistical inference about tumorigenesis should focus on the tumour incidence rate. Unfortunately, in most animal carcinogenicity experiments, tumours are not observable in live animals and censoring of the tumour onset times is informative. In this paper, we propose a Bayesian method for analysing data from such studies. Our approach focuses on the incidence of tumours and accommodates occult tumours and censored onset times without restricting tumour lethality, relying on cause-of-death data, or requiring interim sacrifices. We represent the underlying state of nature by a multistate stochastic process and assume general probit models for the time-specific transition rates. These models allow the incorporation of covariates, historical control data and subjective prior information. The inherent flexibility of this approach facilitates the interpretation of results, particularly when the sample size is small or the data are sparse. We use a Gibbs sampler to estimate the relevant posterior distributions. The methods proposed are applied to data from a US National Toxicology Program carcinogenicity study.     

Attribution of tumour lethality and estimation of the time to onset of occult tumours in the absence of cause-of-death Information

A new statistical approach is developed for estimating the carcinogenic potential of drugs and other chemical substances used by humans. Improved statistical methods are developed for rodent tumorigenicity assays that have interval sacrifices but not cause-of-death data. For such experiments, this paper proposes a nonparametric maximum likelihood estimation method for estimating the distributions of the time to onset of and the time to death from the tumour. The log-likelihood function is optimized using a constrained direct search procedure. Using the maximum likelihood estimators, the number of fatal tumours in an experiment can be imputed. By applying the procedure proposed to a real data set, the effect of calorie restriction is investigated. In this study, we found that calorie restriction delays the tumour onset time significantly for pituitary tumours. The present method can result in substantial economic savings by relieving the need for a case-by-case assignment of the cause of death or context of observation by pathologists. The ultimate goal of the method proposed is to use the imputed number of fatal tumours to modify Peto's International Agency for Research on Cancer test for application to tumorigenicity assays that lack cause-of-death data.     

Tumour incidence, prevalence and lethality estimation in the absence of cause-of-death information

Summary.  A Bayesian intensity model is presented for studying a bioassay problem involving interval-censored tumour onset times, and without discretization of times of death. Both tumour lethality and base-line hazard rates are estimated in the absence of cause-of-death information. Markov chain Monte Carlo methods are used in the numerical estimation, and sophisticated group updating algorithms are applied to achieve reasonable convergence properties. This method was tried on the rat tumorigenicity data that have previously been analysed by Ahn, Moon and Kodell, and our results seem to be more realistic.     

Designs for symmetrical parallel line assays obtainable through group divisible designs

A wide class of block designs for symmetrical parallel line assays with even number of doses, obtainable through group divisible designs is considered. Several new designs can thus be obtained using group divisibledesigns. Group divisible designs are also shown to provide a unification of many exisiting designs which follow as special cases.     

A semiparametric nonlinear mixed model approach to phase I profile monitoring

When process data follow a particular curve in quality control, profile monitoring is suitable and appropriate for assessing process stability. Previous research in profile monitoring focusing on nonlinear parametric (P) modeling, involving both fixed and random-effects, was made under the assumption of an accurate nonlinear model specification. Lately, nonparametric (NP) methods have been used in the profile monitoring context in the absence of an obvious linear P model. This study introduces a novel technique in profile monitoring for any nonlinear and auto-correlated data. Referred to as the nonlinear mixed robust profile monitoring (NMRPM) method, it proposes a semiparametric (SP) approach that combines nonlinear P and NP profile fits for scenarios in which a nonlinear P model is adequate over part of the data but inadequate of the rest. These three methods (P, NP, and NMRPM) account for the auto-correlation within profiles and treats the collection of profiles as a random sample with a common population. During Phase I analysis, a version of Hotelling’s T² statistic is proposed for each approach to identify abnormal profiles based on the estimated random effects and obtain the corresponding control limits. The performance of the NMRPM method is then evaluated using a real data set. Results reveal that the NMRPM method is robust to model misspecification and performs adequately against a correctly specified nonlinear P model. Control charts with the NMRPM method have excellent capability of detecting changes in Phase I data with control limits that are easily computable.     

Linear models analysis of small samples of categorized ordinal response data

A study of the small sample properties of linear models analyses of contingency tables with ordered response categories is presented. Analytical and empirical study of the linear models approach of Williams and Grizzle suggests that modification would improve its performance in small sample situations. Subsequent simulation studies of the modified procedure show that for small samples it has acceptable performance with respect to Type I errors and is robust with respect to response distribution form and inequality among the sample sizes in the contingency table.     

Cost-cautious designs for confirmatory bioassay

Confirmatory bioassay experiments take place in late stages of the drug discovery process when a small number of compounds have to be compared with respect to their properties. As the cost of the observations may differ considerably, the design problem is well specified by the cost of compound used rather than by the number of observations. We show that cost-efficient designs can be constructed using useful properties of the minimum support designs. These designs are particularly suited for studies where the parameters of the model to be estimated are known with high accuracy prior to the experiment, although they prove to be robust against typical inaccuracies of these values. When the parameters of the model can only be specified with ranges of values or by a probability distribution, we use a Bayesian criterion of optimality to construct the required designs. Typically, the number of their support points depends on the prior knowledge for the model parameters. In all cases we recommend identifying a set of designs with good statistical properties but different potential costs to choose from.     

Reducing Conservatism in Risk Estimation for Mixtures of Carcinogens

The excess cancer risk that might result from exposure to a mixture of chemical carcinogens usually must be estimated using data from experiments conducted with individual chemicals. In estimating such risk, it is commonly assumed that the total risk due to the mixture is the sum of the risks of the individual components, provided that the risks associated with individual chemicals at levels present in the mixture are low. This assumption, while itself not necessarily conservative, has led to the conservative practice of summing individual upper-bound risk estimates in order to obtain an upper bound on the total excess cancer risk for a mixture. Less conservative procedures are described here and are illustrated for the case of a mixture of four carcinogens.