Dose addition and the isobole method as approaches for predicting the cumulative effect of non-interacting chemicals: a critical evaluation.

The prediction of the effect of cumulative exposure to similarly acting chemicals is commonly done by dose addition, such as in the relative potency factor approach. This can only be done under the assumption of zero interaction between the chemicals. The related, but not equivalent, isobole method is the most common criterion to judge whether interactions between similarly acting chemicals have taken place in a mixture experiment. Many who apply this latter method assume that it is applicable to any combination of substances, regardless of the shape of the dose-response curves of the individual substances or their underlying mechanism of action. Proponents commonly refer to the work of Berenbaum, who claimed to have proven the general applicability of the isobole method based on zero interaction. In this article, we argue that his argumentation is not generally valid. We further demonstrate that the isobole method, just like dose addition, has limited applicability. Using a physiologically based mathematical model, we provide a theoretical example of a combination of chemicals with zero interaction where the isobole method would result in the decision that they do interact. We discuss the implications for research focusing on detecting or defining interactions, and for the prediction of effects from combined exposures assuming zero interaction

Dose addition and the isobole method as approaches for predicting the cumulative effect of non-interacting chemicals: a critical evaluation.

The prediction of the effect of cumulative exposure to similarly acting chemicals is commonly done by dose addition, such as in the relative potency factor approach. This can only be done under the assumption of zero interaction between the chemicals. The related, but not equivalent, isobole method is the most common criterion to judge whether interactions between similarly acting chemicals have taken place in a mixture experiment. Many who apply this latter method assume that it is applicable to any combination of substances, regardless of the shape of the dose-response curves of the individual substances or their underlying mechanism of action. Proponents commonly refer to the work of Berenbaum, who claimed to have proven the general applicability of the isobole method based on zero interaction. In this article, we argue that his argumentation is not generally valid. We further demonstrate that the isobole method, just like dose addition, has limited applicability. Using a physiologically based mathematical model, we provide a theoretical example of a combination of chemicals with zero interaction where the isobole method would result in the decision that they do interact. We discuss the implications for research focusing on detecting or defining interactions, and for the prediction of effects from combined exposures assuming zero interaction

The use of sandwich-cultured rat hepatocytes to determine the intrinsic clearance of compounds with different extraction ratios: 7-ethoxycoumarin and warfarin.

Item does not contain fulltextThe application of sandwich-cultured rat hepatocytes for the identification of the hepatic intrinsic clearance of compounds with widely varying extraction ratios was investigated. We previously showed the applicability of this in vitro system, in combination with a model describing molecular diffusion, hepatocyte/medium partition, and nonsaturated metabolism, which resulted in a successful identification of this parameter for tolbutamide. This approach is further validated using the compounds 7-ethoxycoumarin and warfarin, covering a 100-fold range of extraction ratios. Clearance of these two substrates could be reliably determined, but only if the depletion of the parent compound in medium as well as in the hepatocyte sandwich was measured. Sensitivity analyses showed that the time course of depletion of the parent compound in medium, especially for warfarin, is insensitive to the partition and diffusion parameter values, whereas depletion in the hepatocyte sandwich was far more sensitive. When varying the volumes of collagen in the sandwich culture, it appears that the most reliable kinetic parameters could be obtained by fitting the data with the smaller collagen volume and that these parameters obtained from fitting to data of the larger volumes generally cannot be verified satisfactorily with the data of the smaller volumes. The values of hepatic clearance that were obtained after extrapolation of the intrinsic clearance to the hepatic clearance from blood were comparable within a factor of 2 to hepatic clearance data in the literature. This indicates that this sandwich culture and modeling system can be applied for the identification of the hepatic intrinsic clearance rate of the total range from low to high clearance compounds

Toxicodynamic analysis of the inhibition of isolated human acetylcholinesterase by combinations of methamidophos and methomyl in vitro.

The applicability of dose addition to combinations of OP-esters and carbamates has been questioned based on theoretical considerations, but these have not been well supported by experimental findings. In the present study, the inhibition of AChE by combinations of methamidophos (an OP-ester) and methomyl (a carbamate) was examined in vitro. AChE inhibition was measured by the Ellman assay. We addressed the question of interaction between the OP-ester and carbamate by a toxicodynamic (TD) model reflecting the mechanism of action of the individual chemicals, without incorporating any interactions between them. The model was extended by including the experimental actions in the Ellman assay to correct for the difference in reactivation rates between phosphorylated and carbamylated AChE, which caused a bias in the observations from the assay. This zero-interactive TD model described the observations well, indicating that the OP-ester and carbamate did not interact. The applicability of dose addition was further explored by applying dose addition to the predicted inhibition by the TD model. Despite the differences in dynamics between methamidophos and methomyl, their dose-response curves were close to parallel, and dose addition gave a reasonably accurate prediction of the combined effects

Facilitated transport of polychlorinated biphenyls and polybrominated diphenyl ethers by dissolved organic matter.

The exchange rate of hydrophobic organic chemicals between the aqueous phase and a sorbent (e.g., soil, organism, passive sampler) is relevant for distribution processes between environmental compartments, including organisms. Dissolved phases such as humic acids, proteins, and surfactants can affect the transfer of such chemicals between the aqueous and sorbent phases by sorption and desorption processes. In this study, the desorption of polychlorinated biphenyls and polybrominated diphenyl ethers from a polymer phase to an aqueous medium was monitored at different humic acid concentrations. The rate of release of the chemical by the polymer phase demonstrates thatthe chemical sorbed to dissolved humic acid contributed significantly to the total mass transfer when the affinity for the humic acid was sufficiently high. This illustrates that environmentally relevant humic acid concentrations can facilitate transport of hydrophobic organic chemicals. The consequences of these facilitated transport mechanisms for uptake into passive samplers are discussed, in particular in situations where equilibration is very slow or when exposure varies in time or space

Toxicodynamic analysis of the combined cholinesterase inhibition by paraoxon and methamidophos in human whole blood.

Theoretical work has shown that the isobole method is not generally valid as a method for testing the absence or presence of interaction (in the biochemical sense) between chemicals. The present study illustrates how interaction can be tested by fitting a toxicodynamic model to the results of a mixture experiment. The inhibition of cholinesterases (ChE) in human whole blood by various dose combinations of paraoxon and methamidophos was measured in vitro. A toxicodynamic model describing the processes related to both OPs in inhibiting AChE activity was developed, and fit to the observed activities. This model, not containing any interaction between the two OPs, described the results from the mixture experiment well, and it was concluded that the OPs did not interact in the whole blood samples. While this approach of toxicodynamic modeling is the most appropriate method for predicting combined effects, it is not rapidly applicable. Therefore, we illustrate how toxicodynamic modeling can be used to explore under which conditions dose addition would give an acceptable approximation of the combined effects from various chemicals. In the specific case of paraoxon and methamidophos in whole blood samples, it was found that dose addition gave a reasonably accurate prediction of the combined effects, despite considerable difference in some of their rate constants, and mildly non-parallel dose-response curves. Other possibilities of validating dose-addition using toxicodynamic modeling are briefly discussed

Clearance and clearance inhibition of the HIV-1 protease inhibitors ritonavir and saquinavir in sandwich-cultured rat hepatocytes and rat microsomes.

The metabolism and active transport of ritonavir and saquinavir were studied using sandwich-cultured rat hepatoyctes and rat liver microsomes. For ritonavir four comparable metabolites were observed in the sandwich-culture and in microsomes. For saquinavir eight metabolites were observed in sandwich-culture and 14 different metabolites in microsomes. Ketoconazole did not affect the metabolism of ritonavir in sandwich-culture or microsomes and slightly inhibited the metabolism of saquinavir in sandwich-culture. This inhibition resulted in a different metabolite profile for saquinavir in microsomes. Ritonavir had a pronounced inhibiting effect on the metabolism of saquinavir and affected the hydroxylation of 6beta-testosterone negatively. In the active transport studies, cyclosporin A and PSC833 enhanced the metabolism of ritonavir, suggesting that ritonavir is normally excreted into the bile canaliculi. Verapamil, showed no effect on the metabolism of ritonavir. The intrinsic clearance was estimated at 1.65 and 67.5 microl/min/1 x 10(6) cells and the hepatic metabolism clearance at 0.017 and 6.83ml/min/SRW for ritonavir and saquinavir respectively. In conclusion, for saquinavir the metabolism rate and the amount of metabolites produced was higher than for ritonavir. Ritonavir had a strong inhibitory effect on the metabolism of saquinavir and seemed to be excreted into the bile

Disposition of smoked cannabis with high [Delta]9-tetrahydrocannabinol content: A kinetic model.

Introduction No model exists to describe the disposition and kinetics of inhaled cannabis containing a high THC dose. We aimed to develop a kinetic model providing estimates of the THC serum concentrations after smoking cannabis cigarettes containing high THC doses (up to 69 mg THC).Methods Twenty-four male non-daily cannabis users smoked cannabis cigarettes containing 29.3 mg, 49.1 mg, and 69.4 mg THC. Blood samples were collected over a period of 0-8 h and serum THC concentrations were measured. A two-compartment open model was fitted on the individual observed data.Results Large inter-individual variability was observed in the pharmacokinetic parameters. The median pharmacokinetic parameters generated by the model were Cmax = 175 ng/mL, Tmax = 14 min, and AUC0-8h = 8150 ng × min/mL for the 69.4 mg THC dose. Median model results show an almost linear dose response relation for Cmax/Dose = 2.8 × 10- 6/mL and AUC0-8h/Dose = 136 × 10- 6 min/mL. However, for increasing dose level, there was a clear decreasing trend: Cmax/Dose = 3.4, 2.6 and 2.5 × 10- 6/mL and AUC0-8h/Dose = 157, 133 and 117 × 10- 6 min/mL for the 29.3, 49.1 and 69.4 mg dose, respectively. Within the restriction of 8 h of observation, the apparent terminal half life of THC was 150 min.Conclusion The model offers insight into the pharmacokinetics of THC in recreational cannabis users smoking cannabis containing high doses of THC mixed with tobacco. The model is an objective method for providing serum THC concentrations up to 8 h after smoking cannabis with a high THC content (up to 23%)