Toward a biologically based dose-response model for developmental toxicity of 5-fluorouracil in the rat: acquisition of experimental data.

Biologically based dose-response (BBDR) models represent an emerging approach to improving the current practice of human health-risk assessment. The concept of BBDR modeling is to incorporate mechanistic information about a chemical that is relevant to the expression of its toxicity into descriptive mathematical terms, thereby providing a quantitative model that will enhance the ability for low-dose and cross-species extrapolation. Construction of a BBDR model for developmental toxicity is particularly complicated by the multitude of possible mechanisms. Thus, a few model assumptions were made. The current study illustrates the processes involved in selecting the relevant information for BBDR modeling, using an established developmental toxicant, 5-fluorouracil (5-FU), as a prototypic example. The primary BBDR model for 5-FU is based on inhibition of thymidylate synthetase (TS) and resultant changes in nucleotide pools, DNA synthesis, cell-cycle progression, and somatic growth. A single subcutaneous injection of 5-FU at doses ranging from 1 to 40 mg/kg was given to pregnant Sprague-Dawley rats at gestational day 14; controls received saline. 5-FU was absorbed rapidly into the maternal circulation, and AUC estimates were linear with administered doses. We found metabolites of 5-FU directly incorporated into embryonic nucleic acids, although the levels of incorporation were low and lacked correlation with administered doses. On the other hand, 5-FU produced dose-dependent inhibition of thymidylate synthetase in the whole embryo, and recovery from enzyme inhibition was also related to the administered dose. As a consequence of TS inhibition, embryonic dTTP and dGTP were markedly reduced, while dCTP was profoundly elevated, perhaps due to feedback regulation of intracellular nucleotide pools. The total contents of embryonic macromolecules (DNA and protein) were also reduced, most notably at the high doses. Correspondingly, dose-related reductions of fetal weight were seen as early as GD 15, and these deficits persisted for the remainder of gestation. These detailed dose-response parameters involved in the expression of 5-FU developmental toxicity were incorporated into mathematical terms for BBDR modeling. Such quantitative models should be instrumental to the improvement of high-to-low dose and cross-species extrapolation in health-risk assessment.

Toward a biologically based dose-response model for developmental toxicity of 5-fluorouracil in the rat: a mathematical construct.

Biologically based dose-response (BBDR) models comprise one way to incorporate mechanistic information into a dose-response assessment to be used for risk assessments. The chemotherapeutic drug 5-fluorouracil (5-FU) has been used as a prototypic compound for the construction of a BBDR model for developmental toxicity. Previous work has provided data and a general mechanistic framework for the developmental toxicity of 5-FU when it was administered to pregnant rats subcutaneously on gestation day 14. In this paper, a mathematical model relating maternally administered treatment with 5-FU to embryonal thymidylate synthetase inhibition and thymidylate synthetase inhibition to various measures of deoxyribonucleotide triphosphate (dNTP) pool perturbation is developed, and parameters are estimated using the data collected. The strategy used was to develop semi-empirical submodels for each of the intervening steps, and to estimate model parameters from previously described data. The models developed predict that there is no practical threshold for dNTP pool perturbation; that is, even minimal doses of 5-FU should result in some perturbation of dNTP pools. In particular, the relationship between dNTP pool perturbation and fetal weight deficit suggests that if there is a biological threshold for the effect of 5-FU on fetal weight, the responsible repair or compensation mechanism must be downstream of dNTP pool perturbation, and saturable at 5-FU doses lower than 10 mg/kg (the lowest dose examined for developmental effects in these studies).

Prioritization of NTP reproductive toxicants for field studies.

Population studies that evaluate human reproductive impairment are time consuming, expensive, logistically difficult, and with limited resources must be prioritized to effectively prevent the adverse health effects in humans. Interactions among health scientists, unions, and industry can serve to identify populations exposed to potential hazards and develop strategies to evaluate and apply appropriate controls. This report describes a systematic method for prioritizing chemicals that may need human reproductive health field studies. Rodent reproductive toxicants identified from the National Toxicology Program (NTP) Reproductive Assessment by Continuous Breeding (RACB) protocol were prioritized on the basis of potency of toxic effect and population at risk. This model for prioritization links NTP findings with data from the National Occupational Exposure Survey (NOES) and the Hazardous Substance Data Base (HSDB) or the High Production Volume Chemical Database (HPVC) to prioritize chemicals for their potential impact on worker populations. The chemicals with the highest priority for field study were: dibutyl phthalate, boric acid, tricresyl phosphate, and N, N-dimethylformamide.

Evaluation of biologically based dose-response modeling for developmental toxicity: a workshop report.

Biologically based dose-response (BBDR) modeling represents a novel approach for quantitative assessment of health risk by incorporating pharmacokinetic and pharmacodynamic characteristics of a chemical and by relating the immediate cellular responses to a cascade of aberrant biological actions that leads to detectable adverse outcomes. The quantitative relationship of each of the intervening events can be described in mathematical forms that are amenable for adjustment and extrapolation over a range of doses and across species. A team of investigators at the Reproductive Toxicology Division of the U.S. Environmental Protection Agency has explored the feasibility of BBDR modeling by examining the developmental toxicity of a known teratogen, 5-fluorouracil. A panel of researchers from academic and industrial laboratories, biomathematical modelers, and risk assessment scientists was convened in a workshop to evaluate the approaches undertaken by the EPA team and to discuss the future prospects of BBDR modeling. This report summarizes the lessons learned from one approach to BBDR modeling and comments from the panelists: while it is possible to incorporate mechanistic information into quantitative dose-response models for the assessment of health risks, the process is enormously data-intensive and costly; in addition, the confidence of the model is directly proportional to our current understanding of basic biology and can be enhanced only through the ongoing novel discoveries. More importantly, the extent of "uncertainty" (inherent with the default assumptions associated with the NOAEL or benchmark approach) reducible by BBDR modeling requires further scrutiny and comparison.

Endocrine-disrupting chemicals: prepubertal exposures and effects on sexual maturation and thyroid activity in the female rat. A focus on the EDSTAC recommendations.

In 1996, the US Environmental Protection Agency was given a mandate by Congress to develop a screening program that would evaluate whether variously identified compounds could affect human health by mimicking or interfering with normal endocrine regulatory functions. Toward this end, the Agency chartered the Endocrine Disruptor Screening and Testing Advisory Committee in October of that year that would serve to recommend a series of in vitro and in vivo protocols designed to provide a comprehensive assessment of a chemical's potential endocrine-disrupting activity. A number of these protocols have undergone subsequent modification by EPA, and this review focuses specifically on the revised in vivo screening procedure recommended under the title Research Protocol for Assessment of Pubertal Development and Thyroid Function in Juvenile Female Rats. Background literature has been provided that summarizes what is currently known about pubertal development in the female rat and the influence of various forms of pharmaceutical and toxicological insult on this process and on thyroid activity. Finally, a section is included that discusses technical issues that should be considered if the specified pubertal endpoints are to be measured and successfully evaluated.

Overview of endocrine disruptor research activity in the United States.

The issue of whether environmental contaminants are inducing adverse health effects in humans and wildlife via interaction with endocrine systems has gained increasing interest during the 1990s. Endocrine disruption is one of the highest priority research topics for the US EPA, and a detailed research strategy has been developed to guide the placement of resources over the next several years. To address the deficiency of testing guidelines in detecting and characterizing damage mediated by interaction with the endocrine system, EPA has issued new multi generation testing guidelines. The new endpoints for monitoring pubertal development, semen quality, and estrous cyclicity will better enable determination of the affected sex, target organ, and life stage following exposure throughout the life cycle. Another major area of effort within EPA is the development of an endocrine disruptor screening program in response to passage of the Food Quality Protection Act of 1996. The current status of these efforts is described. On the federal level, endocrine disruption is one of the five priority research areas for the Committee on the Environment and Natural Resources (CENR) within the Executive Office of President. The CENR has developed a framework to assess research needs for endocrine disruptors, inventoried existing efforts of the federal government (nearly 400 projects were identified as active in FY96), and prioritized additional research needs based upon the needs and gaps in current efforts. It is clear that a great deal of research is underway to clarify the validity of the endocrine disruptor hypothesis and to determine the breadth of chemicals that pose a risk to the endocrine system. The degree of forward research planning and coordination across many organizations should ensure that sufficient data will be available within the next few years to allow a rigorous weight of evidence evaluation that is needed to bring together diverse types of information to make informed decisions regarding risks to humans and wildlife.

Quantitative mechanistically based dose-response modeling with endocrine-active compounds.

A wide range of toxicity test methods is used or is being developed for assessing the impact of endocrine-active compounds (EACs) on human health. Interpretation of these data and their quantitative use in human and ecologic risk assessment will be enhanced by the availability of mechanistically based dose-response (MBDR) models to assist low-dose, interspecies, and (italic)in vitro(/italic) to (italic)in vivo(/italic) extrapolations. A quantitative dose-response modeling work group examined the state of the art for developing MBDR models for EACs and the near-term needs to develop, validate, and apply these models for risk assessments. Major aspects of this report relate to current status of these models, the objectives/goals in MBDR model development for EACs, low-dose extrapolation issues, regulatory inertia impeding acceptance of these approaches, and resource/data needs to accelerate model development and model acceptance by the research and the regulatory community.

The U.S. federal framework for research on endocrine disruptors and an analysis of research programs supported during fiscal year 1996.

The potential health and ecological effects of endocrine disrupting chemicals has become a high visibility environmental issue. The 1990s have witnessed a growing concern, both on the part of the scientific community and the public, that environmental chemicals may be causing widespread effects in humans and in a variety of fish and wildlife species. This growing concern led the Committee on the Environment and Natural Resources (CENR) of the National Science and Technology Council to identify the endocrine disruptor issue as a major research initiative in early 1995 and subsequently establish an ad hoc Working Group on Endocrine Disruptors. The objectives of the working group are to 1) develop a planning framework for federal research related to human and ecological health effects of endocrine disrupting chemicals; 2) conduct an inventory of ongoing federal research programs; and 3) identify research gaps and develop a coordinated interagency plan to address priority research needs. This communication summarizes the activities of the federal government in defining a common framework for planning an endocrine disruptor research program and in assessing the status of the current effort. After developing the research framework and compiling an inventory of active research projects supported by the federal government in fiscal year 1996, the CENR working group evaluated the current federal effort by comparing the ongoing activities with the research needs identified in the framework. The analysis showed that the federal government supports considerable research on human health effects, ecological effects, and exposure assessment, with a predominance of activity occurring under human health effects. The analysis also indicates that studies on reproductive development and carcinogenesis are more prevalent than studies on neurotoxicity and immunotoxicity, that mammals (mostly laboratory animals) are the main species under study, and that chlorinated dibenzodioxins and polychlorinated biphenyls are the most commonly studied chemical classes. Comparison of the inventory with the research needs should allow identification of underrepresented research areas in need of attention.

Effects of boric acid on axial skeletal development in rats.

Prenatal exposure to elevated levels of boric acid (BA) causes reduced incidences of supernumerary ribs and shortening/absence of the 13th rib in multiple laboratory species. To explore this further, Sprague-Dawley rats received 500 mg/kg b.i.d. on gestation days (gd) 5-9, 6-9, 6-10, or on single days between gd 6 and 11 (plug day = gd 0); gd-21 fetuses were stained for skeletal examination. Following multiday exposures, malformations of the axial skeleton involved the head, sternum, ribs, and vertebrae. Shortening/absence of the 13th rib was seen particularly in the gd 5-9 and 6-10 exposure groups. Although most groups exposed on single days were generally unaffected, about 90% of the gd-9 exposed fetuses had only six cervical vertebrae; the deficient region was usually C3-C5. In contrast, gd-10 treatment caused agenesis of a thoracic/lumbar vertebra in over 60% of the fetuses; the deficient region was usually T11. For 13-ribbed fetuses, the length of rib 13 was shortened compared to controls. Postnatal assessment suggested increased mortality for gd-10 exposed pups. Embryos in culture showed reduced development when exposed to BA for 48 h. These findings demonstrate the critical periods for axial development in the rat and provide an experimental model for the study of homeotic shifts.

Effect of dosing vehicle on the developmental toxicity of bromodichloromethane and carbon tetrachloride in rats.

Several halocarbons have been shown to cause full-litter resorption (FLR) in Fischer-344 rats when administered orally in corn oil. Since halocarbons often occur as contaminants of drinking water, we sought to determine the influence of the vehicle, aqueous versus lipid, on the developmental toxicity of two of these agents. In separate assays, bromodichloromethane (BDCM) and carbon tetrachloride (CCl4) were administered by gavage to Fischer-344 rats on gestation days (GD) 6-15 at 0, 25, 50, or 75 mg/kg/day in either corn oil or an aqueous vehicle containing 10% Emulphor EL-620. Dams were allowed to deliver and the litters were examined postnatally. Uteri of females that did not deliver were stained with 10% ammonium sulfide to detect FLR. Effects of both agents on maternal weight gain were slightly more pronounced in the aqueous vehicle at lower doses, but at the highest dose, CCl4 was more maternally toxic in corn oil. Developmentally, both agents caused FLR at 50 and 75 mg/kg in both vehicles. At 75 mg/kg, dams receiving corn oil had significantly higher rates of FLR (83% for BDCM, 67% for CCl4) compared to their aqueous-vehicle counterparts (21% for BDCM, 8% for CCl4). Blood concentrations of BDCM following GD-6 gavage revealed a shorter elimination half-life in the aqueous dosing vehicle (2.7 h) compared to the oil vehicle (3.6 h). Benchmark doses of CCl4 were similar for the oil (18.9 mg/kg) and aqueous (14.0 mg/kg) vehicles. For BDCM, the corn oil vehicle yielded a less conservative (i.e., higher) value (39.3 mg/kg) than the aqueous vehicle (11.3 mg/kg), reflecting different confidence intervals around the estimated 5%-effect dose levels.

Critical period of carbon tetrachloride-induced pregnancy loss in Fischer-344 rats, with insights into the detection of resorption sites by ammonium sulfide staining.

Several low-molecular weight halocarbons have been shown to cause full-litter resorption (FLR), i.e., pregnancy loss, in Fischer-344 rats treated during organogenesis. To determine periods of gestation sensitive to acute exposure, a single dose of 150 mg carbon tetrachloride (CCl4)/kg was administered on gestation day (GD) 6, 7, 8, 10, or 12. Fetuses were delivered by cesarean section on GD 20. Non-gravid uteri were examined for resorption sites, placed in 10% ammonium sulfide, and re-examined for stained resorption sites approximately 1 and 4.5 hr later. FLR was seen in 4% (1/27) of control dams and 36% (4/11), 54% (7/13), 72% (18/25), 54% (7/13), and 0% (0/12) of dams treated on GD 6, 7, 8, 10, and 12, respectively. Ammonium sulfide staining clearly yielded a more accurate account of the incidence of FLR. The technique was most effective when the staining period was extended to 4.5 hr, as two cases of FLR were revealed that had been undetected after 1 hr of staining. For dams with FLR, staining was required to detect resorption sites in all dams treated on GD 6 or 7, most dams treated on GD 8, and one dam treated on GD 10. Fewer implantation sites were detected in the dams treated on GD 6, and the size of the stained resorption sites increased as the day of treatment was delayed. These findings demonstrate a relationship between the time of toxicant exposure and the size and detectability of resorption sites near term, suggesting that the size of the resorption site may reliably reflect the time of embryonic death. Treatment on GD 8 caused the highest incidence of FLR and will be used in subsequent mechanistic research.

Nucleoside-mediated mitigation of 5-fluorouracil-induced toxicity in synchronized murine erythroleukemic cells.

5-Fluorouracil (5-FU) is a chemotherapeutic agent known to retard embryonic growth and induce cleft palate and limb deformities. The predominant mechanism underlying its toxic action is thought to be inhibition of thymidylate synthetase (TS), and hence thymidine triphosphate (dTTP) synthesis, resulting in alteration of the balance of deoxynucleotide (dNTP) pools and disruption of DNA synthesis. Indeed, previously we demonstrated retarded cell-cycle progression concurrent with a 60% decrease in TS activity in rat whole embryos following maternal exposure to 40 mg/kg 5-FU on Gestational Day 14 and in the murine erythroleukemic cell (MELC) suspension culture following exposure to 5-25 microM 5-FU for 2 hr. In the study described herein, we used high-performance liquid chromatography (HPLC) to demonstrate in both of these model systems that 5-FU exposure results in similar patterns of dNTP perturbations: a prolonged decrease in dTTP and dGTP levels and an increase in dCTP and dATP. In addition, we used centrifugal elutriation to synchronize MELC in the phases of the cell cycle (G0/G1 and early S) most sensitive to 5-FU to investigate the ability of nucleoside supplementation to mitigate 5-FU-induced toxicity. Our data indicate that following a 2-hr exposure to 5-25 microM 5-FU, supplementation with 1-10 microM thymidine (TdR) for 24 hr partially reverses 5-FU-induced toxicity as evidenced by increased cellular proliferation and cell-cycle progression and amelioration of 5-FU-induced perturbations of protein synthesis and cellular membrane permeability compared to unsupplemented 5-FU-exposed cells. However, TdR concentrations >/=100 microM inhibited growth or were cytotoxic. In comparison, supplementation with 10 microM-10 mM of deoxycytidine (CdR) was not toxic, but effected a dose-dependent recovery from 5-FU-induced toxicity. At 1-100 microM, neither deoxyadenosine nor deoxyguanosine supplementation reduced 5-FU-induced toxicity; at higher concentrations, both purine nucleotides inhibited cell growth. Although these results support the hypothesis that 5-FU disrupts the MELC cell cycle by depleting dTTP (a perturbation that is reversible by TdR supplementation), they also indicate that CdR supplementation offers an additional recovery pathway.

Stereoselective dysmorphogenicity of the enantiomers of the valproic acid analogue 2-N-propyl-4-pentynoic acid (4-yn-VPA): cross-species evaluation in whole embryo culture.

We previously reported the in vitro differential stereoselective dysmorphogenic potential of the R(+) and S(-) enantiomers of 2n-propyl-4-pentynoic acid (4-yn-VPA) in mice. To determine whether this stereoselectivity is species specific, we evaluated the dysmorphogenic potential of these isomers as well as valproic acid (VPA) to gestational day 9 rat embryos using whole embryo culture (WEC). Aqueous solutions of the sodium salts of R-4-yn-VPA, S-4-yn-VPA, 50%R/ 50%S-4-yn-VPA or VPA were added to the culture medium to give 0, 0.075, 0.15, 0.3, 0.6, or 1.2 mmol/L and embryos were evaluated 48 hr later. The S-4-yn-VPA enantiomer gave clear concentration-dependent dysmorphology as well as effects on developmental score, somite number, crown rump length, and head length. Effects on rotation and defects of the neural tube, somites and heart were observed. Embryolethality was observed only at 1.2 mmol/L concentration. The R-4-yn-VPA enantiomer was neither embryo toxic nor dysmorphogenic at any concentration. VPA significantly reduced all parameters and was dysmorphogenic at the highest concentration but was not embryo lethal. The 50/50 mixture of R- and S-isomers appeared to elicit a degree of embryolethality and dysmorphology similar to VPA. The potency order for the four chemicals was S(-) > S(-)/R(+) = VPA > > > R(+), comparable to that observed in mice by either in vivo or in vitro exposure. These data demonstrate that the stereoselective dysmorphology for these enantiomers can be observed across species and is not related to maternal metabolism.

Recent advances in mathematical modeling of developmental abnormalities using mechanistic information.

During the last several years, significant changes in the risk assessment process for developmental toxicity of environmental contaminants have begun to emerge. The first of these changes is the development and beginning use of statistically based dose-response models [the benchmark dose (BMD) approach] that better utilize data derived from existing testing approaches. Accompanying this change is the greater emphasis placed on understanding and using mechanistic information to yield more accurate, reliable, and less uncertain risk assessments. The next stage in the evolution of risk assessment will be the use of biologically based dose-response (BBDR) models that begin to build into the statistically based models factors related to the underlying kinetic, biochemical, and/or physiologic processes perturbed by a toxicant. Such models are now emerging from several research laboratories. The introduction of quantitative models and the incorporation of biologic information into them has pointed to the need for even more sophisticated modifications for which we offer the term embryologically based dose-response (EBDR) models. Because these models would be based upon the understanding of normal morphogenesis, they represent a quantum leap in our thinking, but their complexity presents daunting challenges both to the developmental biologist and the developmental toxicologist. Implementation of these models will require extensive communication between developmental toxicologists, molecular embryologists, and biomathematicians. The remarkable progress in the understanding of mammalian embryonic development at the molecular level that has occurred over the last decade combined with advances in computing power and computational models should eventually enable these as yet hypothetical models to be brought into use.

Endocrine disruptors and reproductive development: a weight-of-evidence overview.

It is clear that the endocrine system presents a number of target sites for the induction of adverse effects by environmental agents (Fig. 1). There are numerous examples demonstrating that reproductive and developmental processes may be exquisitely sensitive to exposure and there are clear effects induced by presumed endocrine-disrupting chemicals in a variety of species. The concerns raised by studies of wildlife and humans place added significance on a better understanding of the myriad of effects attributed to endocrine disruptors. But there remains a large void between the study of relatively high exposure levels used in laboratory settings versus the relatively low levels found in the general environment. It is also equally clear that the term "endocrine disruption' has been applied to situations where the biological basis is far from conclusive. This may be a moot point in situations where populations are experiencing adverse effects on reproduction, but as scientists we must be vigilant of the appropriate use of descriptive terminology, particularly in cases where public awareness and concern are as great as this. Not only is there a need for better test procedures (both in vivo and in vitro) to characterize the potential of environmental agents to disrupt endocrine function in laboratory species, but there is also a need for a more comprehensive understanding of the normal physiological processes associated with reproduction and development in those wildlife species studied. At the same time, obtaining better information on the transport, fate and bioavailability of chemicals released into the environment remains an important but imposing task. The goals of risk characterization are to carefully delineate cause-and-effect relationships, define the dose-response relationships, and determine whether environmental exposures exceed acceptable levels. A concerted research effort is needed to fill the voids in our knowledge and reduce the large uncertainties that exist today. Only then can regulatory actions take place within the confines of legislative mandates, remediation strategies and considerations of international use and transport. Towards this end, the US Environmental Protection Agency (EPA) sponsored two workshops in 1995 (Ankley et al. 1996, Kavlock et al. 1996) at which groups of international scientists began the process of identifying research needs. Similar efforts also took place in several European countries at about the same time (Danish Environmental Protection Agency 1995, Medical Research Council 1995, Umweltbundesamt 1995). More recently, a Working Group on Endocrine Disruptors has been established within the Committee on the Environment and Natural Resources of the US Government's National Science and Technology Council. The objectives of this Working Group are to (1) formulate a framework for identifying research needs related to the health and ecological effects of endocrine-disrupting chemicals; (2) conduct an inventory of on-going federal research programs; and (3) identify research gaps and facilitate a co-ordinated research plan to address them. These efforts were largely completed in the Fall of 1996 and the information will be made available via the Internet (http;@www.cpa.gov/endocrine). The group also plans to work more broadly with other governments and private industry and public interest groups conducting research on this tissue to co-ordinate research and disseminate scientific information. Persons wishing to know more about this effort should contact the authors. The issue of endocrine disruption has raised the consciousness of many researchers, both within and outside the toxicology community, and has attracted considerable public and political interest. We now have the beginnings of international co-operation to identify the most important scientific uncertainties and to dedicate resources to address the critical gaps. (ABSTRACT TRUNCATED)

The workshop on endocrine disrupter research needs: a report.

On April 10-13, 1995 the US EPA sponsored a workshop to develop research needs for endocrine disrupters. Participants were assigned to discussion groups for health effects issues and risk assessment methodologies. The neurotoxicology workgroup identified several chemicals including the PCBs and dioxins that affect nervous system function possibly by acting on the endocrine system during development. The study of endocrine disrupter is confounded by a number of uncertainties, including the presence of chemical mixtures in the environment unclear exposure parameters, poorly understood mechanisms of action, poor dose-response characterization, and uncertain animal-to-human extrapolation. The working group proposed a research strategy to address these uncertainties, which includes initial identification of effects of concern to human and/or wildlife populations and determining whether those effects can be associated with exposure to specific chemicals in the environment and neuroendocrine disruption. Once a problem chemical has been identified and the exposure conditions established, hypothesis-driven research to determine mechanism of action could proceed.

A perspective on the risk assessment process for endocrine-disruptive effects on wildlife and human health.

The topic of EDCs presents significant issues to the risk assessment process. In Table I, we have summarized many of the issues raised above. We have a working definition of an EDC, that provides a starting point for considering what chemicals are of concern. We also have an understanding of the important biological endpoints. Significantly, there are indications that large scale impacts might be occurring in both human and wildlife populations. Should these effects be confirmed and the causative agents identified, we will face difficult risk management decisions to minimize or mitigate the risks. Our present knowledge base, however, suggests that the traditional approach to assessing both noncancer and cancer endpoints will be suitable for toxicity mediated through disruption of endocrine systems, although there is controversy here as well. The fact that many EDCs share a common MOA should, in fact, assist in developing more scientifically defensible risk assessments, given that we must often extrapolate across doses and species in the absence of a mechanistic basis for noncancer effects. As more research focuses on the key uncertainties, we should acquire a better vision of the relative risks that EDCs pose to humans and wildlife.

Research needs for the risk assessment of health and environmental effects of endocrine disruptors: a report of the U.S. EPA-sponsored workshop.

The hypothesis has been put forward that humans and wildlife species adverse suffered adverse health effects after exposure to endocrine-disrupting chemicals. Reported adverse effects include declines in populations, increases in cancers, and reduced reproductive function. The U.S. Environmental Protection Agency sponsored a workshop in April 1995 to bring together interested parties in an effort to identify research gaps related to this hypothesis and to establish priorities for future research activities. Approximately 90 invited participants were organized into work groups developed around the principal reported health effects-carcinogenesis, reproductive toxicity, neurotoxicity, and immunotoxicity-as well as along the risk assessment paradigm-hazard identification, dose-response assessment, exposure assessment, and risk characterization. Attention focused on both ecological and human health effects. In general, group felt that the hypothesis warranted a concerted research effort to evaluate its validity and that research should focus primarily on effects on development of reproductive capability, on improved exposure assessment, and on the effects of mixtures. This report summarizes the discussions of the work groups and details the recommendations for additional research.