Non-monotonic Dose Response Research Articles

Aristolochic acid-induced carcinogenesis examined by ACB-PCR quantification of H-Ras and K-Ras mutant fraction

Aristolochic acid (AA) is a strong cytotoxic nephrotoxin and carcinogen associated with the development of urothelial cancer in humans. AA induces forestomach, kidney and urothelial tract tumours in rats and mice. This study was conducted to characterise AA's carcinogenic mechanism of action and compare allele-specific competitive blocker-polymerase chain reaction (ACB-PCR)-based early detection of carcinogenic effect using two different tumour-relevant endpoints. H-Ras codon 61 CAA→CTA mutation was analysed because it is found in rodent forestomach tumours and A:T→T:A transversion is the predominant mutational specificity induced by AA. K-Ras codon 12 GGT→GAT mutation was analysed because it is a common spontaneous mutation present in various rodent tissues and may be a useful generic biomarker for carcinogenic effect. DNA samples from Big Blue rats treated with 0, 0.1, 1.0 or 10.0 mg AA/kg body weight (bw) by gavage, 5 days/week for 12 weeks were used in ACB-PCR in order to examine the induction of the two specific mutations. A significant dose-dependent induction of H-Ras mutant fraction (MF) was observed in liver and kidney. Statistically significant correlations were observed between AA-induced DNA adduct levels or cII mutant frequencies (previously measured in the same rats) and H-Ras MF measurements. No correlation between AA dose and K-Ras MF was found in liver or kidney, although there was a significant induction of K-Ras mutation in kidneys exposed to 0.1 mg/kg bw AA relative to controls. Thus, the data establish a straightforward dose-related increase in H-Ras MF due to fixation of AA-induced DNA adducts, whereas the common spontaneous K-Ras mutation showed a non-monotonic dose-response, consistent with loss of non-targeted mutation at cytotoxic doses.

The fungicide chlorothalonil is nonlinearly associated with corticosterone levels, immunity, and mortality in amphibians.

Background: Contaminants have been implicated in declines of amphibians, a taxon with vital systems similar to those of humans. However, many chemicals have not been thoroughly tested on amphibians or do not directly kill them.Objective: Our goal in this study was to quantify amphibian responses to chlorothalonil, the most commonly used synthetic fungicide in the United States.Methods: We reared Rana sphenocephala (southern leopard frog) and Osteopilus septentrionalis (Cuban treefrog) in outdoor mesocosms with or without 1 time (1×) and 2 times (2×) the expected environmental concentration (EEC) of chlorothalonil (~ 164 μg/L). We also conducted two dose–response experiments on O. septentrionalis, Hyla squirella (squirrel treefrog), Hyla cinerea (green treefrog), and R. sphenocephala and evaluated the effects of chlorothalonil on the stress hormone corticosterone.Results: For both species in the mesocosm experiment, the 1× and 2× EEC treatments were associated with > 87% and 100% mortality, respectively. In the laboratory experiments, the approximate EEC caused 100% mortality of all species within 24 hr; 82 μg/L killed 100% of R. sphenocephala, and 0.0164 μg/L caused significant tadpole mortality of R. sphenocephala and H. cinerea. Three species showed a nonmonotonic dose response, with low and high concentrations causing significantly greater mortality than did intermediate concentrations or control treatments. For O. septentrionalis, corticosterone exhibited a similar nonmonotonic dose response and chlorothalonil concentration was inversely associated with liver tissue and immune cell densities (< 16.4 μg/L).Conclusions: Chlorothalonil killed nearly every amphibian at the approximate EEC; at concentrations to which humans are commonly exposed, it increased mortality and was associated with elevated corticosterone levels and changes in immune cells. Future studies should directly quantify the effects of chlorothalonil on amphibian populations and human health.

Lead Doesn’t Spare the Rod: Low-Level Exposure Supercharges Retinal Cell Production in Mice

Low-level gestational lead exposure has been shown to increase the electrical response of the rod signaling pathway in the retinas of children, monkeys, and rats, which could in turn contribute to retinal disease. Now researchers demonstrate the phenomenon underlying this effect: increased proliferation of retinal progenitor cells, which give rise to functionally differentiated retinal cells that sense and transmit visual information [EHP 119(1):71–77; Giddabasappa et al.]. Using a previously described mouse model of low-level gestational lead exposure, the researchers set out to test the hypothesis that such exposure selectively increases rod photoreceptors and bipolar cells in the rod signaling pathway. (The rod signaling pathway detects gradations of light, as opposed to the cone signaling pathways, which detect colors.) Female mice were given water containing varying concentrations of lead: 0 ppm (control), 27 ppm (“low” dose), 55 ppm (“moderate” dose), or 109 ppm (“high” dose). The exposures were administered for 2 weeks before mating, during pregnancy, and through postnatal day 10—a model for the human gestation period. On postnatal day 10, unspiked water replaced the water–lead mixtures for all groups. The adult mammalian retina consists of six types of neurons and a Muller glial cell. These cell types develop in one of two distinct phases: primarily in utero (“early-born”) or primarily after birth (“late-born”). In examining controls and exposed mice at postnatal day 60, the researchers found that late-born rod photoreceptors and rod and cone bipolar cells increased by 16–30% in exposed offspring, whereas Muller glial cells (also classified as late-born retinal cells) did not increase. Low and moderate lead doses showed the greatest effects. Gestational lead exposure also increased and prolonged retinal progenitor cell proliferation but did not alter developmental apoptosis (programmed cell death), indicating that the higher numbers of rods and bipolar cells were due to increased production, not decreased apoptosis. These results demonstrate that gestational lead exposure resulting in blood lead levels of 10 μg/dL alters retinal development by selectively promoting the development of rod photoreceptor cells and bipolar cells. The authors speculate that the increased number of rods and bipolar cells in the lead-exposed animals could accelerate age-related retinal degeneration. These nonmonotonic dose–response results raise complex issues for neurotoxicology, risk assessment, public health, and children’s health.

Estrogens from the Outside In: Alkylphenols, BPA Disrupt ERK Signaling in Vitro

Vol. 119, No. 1 NewsOpen AccessEstrogens from the Outside In: Alkylphenols, BPA Disrupt ERK Signaling in Vitro Julia R. Barrett Julia R. Barrett Search for more papers by this author Published:1 January 2011https://doi.org/10.1289/ehp.119-a34bCited by:1AboutSectionsPDF ToolsDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InReddit The body produces estrogens—including estrone (E1), estradiol (E2), and estriol (E3)—that direct reproductive system processes and contribute to the normal function of tissues including the brain, bone, and cardiovascular system. Certain xenoestrogens (estrogenic compounds introduced from outside the body) are suspected of disrupting these activities. In a new study, xenoestrogenic alkylphenols and bisphenol A (BPA) interfered with normal estrogenic signaling in vitro, which suggests they could disrupt normal physiologic function at critical life stages [EHP119(1):104–112; Jeng and Watson].Different estrogen receptors control different functions: receptors in the cell nucleus direct gene transcription, whereas receptors in the cell membrane direct signaling pathways via extracellular signal–regulated kinases (ERKs). ERK-controlled pathways respond to many biochemical stimuli and integrate these signals to direct a cell toward division, differentiation, death, or malignant transformation. The structurally related alkylphenols and BPA interact weakly with nuclear estrogen receptors, but they can have pronounced effects on signaling pathways mediated by estrogen receptors in the cell membrane.In the current study, a rat pituitary cancer cell line was used to study the effect of alkylphenols and BPA on ERK1 and ERK2 activation (measured as phosphorylation), both alone and in combination with each physiologic estrogen. After treatment with each physiologic and environmental estrogen, the researchers measured time-dependent surges in ERK activation. In most cases, E1 and E2 prompted early, intermediate, and late surges in ERK activation at 5, 10–30, and >30 min, respectively; alkylphenols and E3 typically triggered early and late surges. Interestingly, a very low concentration of BPA (10−14 M) yielded a similar two-peak response, but a higher concentration (1 nM) induced a three-peak response like that of E1 and E2. Both BPA concentrations were typical of environmental exposures and, along with ineffective midrange doses, also illustrated the nonmonotonic dose–response relationship characteristic of many estrogenic compounds.When physiologic estrogens and xenoestrogens were combined, the response pattern generally shifted to a single major peak at an intermediate time. Xenoestrogens that caused a strong response when administered alone at a particular point in time or concentration tended to inhibit ERK activation in response to a physiologic estrogen. But at other times or concentrations, the same xenoestrogen might cause a weak response on its own, in which case it would tend to enhance ERK phosphorylation in response to physiologic estrogens.There were exceptions to these general patterns, however, which highlights the need to study effects of individual xenoestrogens at different points in time, at varying concentrations, and in different tissues. The effect of shifts in the patterns of ERK activation are only just beginning to be explored, although it is known that these patterns constitute an important component of information flow within a cell. The correct flow of information is likely to be especially critical during windows of vulnerability that are based in part on life stage.FiguresReferencesRelatedDetailsCited by Park M, Hwang K and Choi K (2011) Diverse animal models to examine potential role(s) and mechanism of endocrine disrupting chemicals on the tumor progression and prevention: Do they have tumorigenic or anti-tumorigenic property?, Laboratory Animal Research, 10.5625/lar.2011.27.4.265, 27:4, (265), . Vol. 119, No. 1 January 2011Metrics About Article Metrics Publication History Originally published1 January 2011Published in print1 January 2011 Financial disclosuresPDF download License information EHP is an open-access journal published with support from the National Institute of Environmental Health Sciences, National Institutes of Health. All content is public domain unless otherwise noted. Note to readers with disabilities EHP strives to ensure that all journal content is accessible to all readers. However, some figures and Supplemental Material published in EHP articles may not conform to 508 standards due to the complexity of the information being presented. If you need assistance accessing journal content, please contact [email protected]. Our staff will work with you to assess and meet your accessibility needs within 3 working days.

Bisphenol A and the non-monotonic dose response: Why environmentally relevant doses pose more danger than pharmacological doses

Bisphenol A and the non-monotonic dose response: Why environmentally relevant doses pose more danger than pharmacological doses

Developmental neurotoxicity study of dietary bisphenol A in Sprague-Dawley rats.

This study was conducted to determine the potential of bisphenol A (BPA) to induce functional and/or morphological effects to the nervous system of F1 offspring from dietary exposure during gestation and lactation according to the Organization for Economic Cooperation and Development and U.S. Environmental Protection Agency guidelines for the study of developmental neurotoxicity. BPA was offered to female Sprague-Dawley Crl:CD (SD) rats (24 per dose group) and their litters at dietary concentrations of 0 (control), 0.15, 1.5, 75, 750, and 2250 ppm daily from gestation day 0 through lactation day 21. F1 offspring were evaluated using the following tests: detailed clinical observations (postnatal days [PNDs] 4, 11, 21, 35, 45, and 60), auditory startle (PNDs 20 and 60), motor activity (PNDs 13, 17, 21, and 61), learning and memory using the Biel water maze (PNDs 22 and 62), and brain and nervous system neuropathology and brain morphometry (PNDs 21 and 72). For F1 offspring, there were no treatment-related neurobehavioral effects, nor was there evidence of neuropathology or effects on brain morphometry. Based on maternal and offspring body weight reductions, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was 75 ppm (5.85 and 13.1 mg/kg/day during gestation and lactation, respectively), with no treatment-related effects at lower doses or nonmonotonic dose responses observed for any parameter. There was no evidence that BPA is a developmental neurotoxicant in rats, and the NOAEL for developmental neurotoxicity was 2250 ppm, the highest dose tested (164 and 410 mg/kg/day during gestation and lactation, respectively).

Pubertal Administration of DEHP Delays Puberty, Suppresses Testosterone Production, and Inhibits Reproductive Tract Development in Male Sprague-Dawley and Long-Evans Rats

Although is clear that exposure to high dosage levels of some phthalates delays the onset of puberty in the male rat, it has been hypothesized that low levels of di(2-ethylhexyl) phthalate (DEHP) accelerate puberty by enhancing testicular androgen synthesis. The current study was designed to determine if the dose response to DEHP was nonmonotonic, as hypothesized. Pubertal administration of DEHP delayed the onset of puberty and reduced androgen-dependent tissue weights in both Long-Evans (LE) and Sprague-Dawley (SD) male rats 300 and 900 mg DEHP/kg/day. These effects were generally of greater magnitude in LE than SD rats. By contrast, alterations in testis histopathology (300 and 900 mg/kg/day) were more severe in SD than in LE rats. Taken together, these results suggest that DEHP may be acting on the pubertal male rat testis via two modes of action; one via the Leydig cells and the other via the Sertoli cells. Treatment with DEHP generally reduced serum testosterone and increased serum luteinizing hormone (LH) levels, demonstrating that the reduction in testosterone was due to the effect of DEHP on the testis and not via an inhibition of LH from hypothalamic-pituitary axis. Testosterone production ex vivo (with and without human chorionic gonadotropin stimulation) was consistently reduced in males at the time of puberty and shortly thereafter. DEHP treatment did not accelerate the age at puberty or enhance testosterone levels at 10 or 100 mg/kg/day in either LE or SD rats, as some have hypothesized. Taken together, these results do not provide any evidence of a nonmonotonic dose response to DEHP during puberty.

Quantitative structure-activity relationships of selected phenols with non-monotonic dose-response curves

Particular non-monotonic dose-response curves of many endocrine disrupting chemicals (EDCs) suggest the existence of diverse toxicity mechanisms at different dose levels. As a result, the biological activities of EDCs cannot be simply exhibited by unique EC50/LD50 values, and the quantitative structure-activity relationship (QSAR) analysis for non-monotonic dose-response relationship becomes an unknown field in the environmental science. In this paper, nine phenols with inverted U-shaped dose-response curves in lymphocyte proliferation test of Carassius auratus were selected. The binding interactions between the phenols and several typical EDCs-related receptors were then explored in a molecular simulation study. The estrogen receptor (ER), androgen receptor (AR), thyroid hormone receptor (TR), bacterial O2 sensing FixL protein (FixL), aryl hydrocarbon receptor (AhR), and the peroxisome proliferator-activated receptor (PPAR) were the target receptors in the study. Linear regression QSAR models for the low and high exposure levels of the compounds were developed separately. The results indicated that the lymphocyte proliferation in the low-dose range might involve ER-mediated process, while the proliferation inhibition in the high dose range was dominated by the acute toxicity of phenols due to receptor occupancy and cell damage.

Influence of Auditory Stimulation Rates on Evoked Potentials during General Anesthesia

The auditory middle-latency response (transient) and the 40-Hz auditory steady state response (ASSR) are modulated by anesthetics. However, the quantitative relation between these evoked responses is difficult to obtain because of technical limitations of the recording methods used to obtain transients at high stimulation rates. This study uses continuous-loop averaging deconvolution to fill this technical gap and to study the relation between the transient and ASSR waveform during general anesthesia. The authors recorded 5- and 40-Hz transients and 40-Hz ASSRs in 13 subjects during general anesthesia. The 5- and 40-Hz transients were used to predict the 40-Hz ASSR by linearly superimposing the transient waveforms. The predicted and recorded ASSRs were analyzed and compared using phasor and Hotelling T(2) analyses. Grand-averaged recordings revealed differences in the early middle-latency peaks between 5- and 40-Hz transients, e.g., the peak P(x) was present only in 5-Hz transient. Only the predicted 40-Hz ASSR derived from the 40-Hz transient matched the actual ASSR. Phasor analysis showed that the early peaks contribute significantly to the steady state waveform, and this explains why 5-Hz transient does not predict the 40-Hz ASSR. Oscillations in both the 5- and 40-Hz transients were observed during anesthesia. The 40-Hz ASSR represents a composite waveform and arises when transient waveforms elicited with a 40-Hz stimulation rate are overlapped and superimposed. During general anesthesia, the morphology of the transient is dependent on the rate of stimulus presentation. The composite nature of the ASSR may explain nonmonotonic anesthetic dose-response relations observed by others.

Membrane estrogen receptor-α-mediated nongenomic actions of phytoestrogens in GH&lt;sub&gt;3&lt;/sub&gt;/B&lt;sub&gt;6&lt;/sub&gt;/F&lt;sub&gt;10&lt;/sub&gt; pituitary tumor cells

BackgroundEstradiol (E2) mediates various intracellular signaling cascades from the plasma membrane via several estrogen receptors (ERs). The pituitary is an estrogen-responsive tissue, and we have previously reported that E2 can activate mitogen-activated protein kinases (MAPKs) such as ERK1/2 and JNK1/2/3 in the membrane ERα (mERα)-enriched GH3/B6/F10 rat pituitary tumor cell line. Phytoestrogens are compounds found in plants and foods such as soybeans, alfalfa sprouts, and red grapes. They are structurally similar to E2 and share a similar mechanism of action through their binding to ERs. Phytoestrogens bind to nuclear ERs with a much lower affinity and therefore are less potent in mediating genomic responses. However, little is known about their ability to act via mERs to mediate nongenomic effects.MethodsTo investigate the activation of different nongenomic pathways, and determine the involvement of mERα, we measured prolactin (PRL) release by radio-immunoassay, MAPK activations (ERK1/2 and JNK1/2/3) via a quantitative plate immunoassay, and intracellular [Ca2+] by Fura-2 fluorescence imaging in cells treated with E2 or four different phytoestrogens (coumestrol, daidzein, genistein, and trans-resveratrol).ResultsCoumesterol and daidzein increased PRL release similar to E2 in GH3/B6/F10 cells, while genistein and trans-resveratrol had no effect. All of these compounds except genistein activated ERK1/2 signaling at 1–10 picomolar concentrations; JNK 1/2/3 was activated by all compounds at a 100 nanomolar concentration. All compounds also caused rapid Ca2+ uptake, though in unique dose-dependent Ca2+ response patterns for several aspects of this response. A subclone of GH3 cells expressing low levels of mERα (GH3/B6/D9) did not respond to any phytoestrogen treatments for any of these responses, suggesting that these nongenomic effects were mediated via mERα.ConclusionPhytoestrogens were much more potent in mediating these nongenomic responses (activation of MAPKs, PRL release, and increased intracellular [Ca2+]) via mERα than was previously reported for genomic responses. The unique non-monotonic dose responses and variant signaling patterns caused by E2 and all tested phytoestrogens suggest that complex and multiple signaling pathways or binding partners could be involved. By activating these different nongenomic signaling pathways, phytoestrogens could have significant physiological consequences for pituitary cell functions.

Bisphenol-A and the great divide: a review of controversies in the field of endocrine disruption.

In 1991, a group of 21 scientists gathered at the Wingspread Conference Center to discuss evidence of developmental alterations observed in wildlife populations after chemical exposures. There, the term "endocrine disruptor" was agreed upon to describe a class of chemicals including those that act as agonists and antagonists of the estrogen receptors (ERs), androgen receptor, thyroid hormone receptor, and others. This definition has since evolved, and the field has grown to encompass hundreds of chemicals. Despite significant advances in the study of endocrine disruptors, several controversies have sprung up and continue, including the debate over the existence of nonmonotonic dose response curves, the mechanisms of low-dose effects, and the importance of considering critical periods of exposure in experimental design. One chemical found ubiquitously in our environment, bisphenol-A (BPA), has received a tremendous amount of attention from research scientists, government panels, and the popular press. In this review, we have covered the above-mentioned controversies plus six additional issues that have divided scientists in the field of BPA research, namely: 1) mechanisms of BPA action; 2) levels of human exposure; 3) routes of human exposure; 4) pharmacokinetic models of BPA metabolism; 5) effects of BPA on exposed animals; and 6) links between BPA and cancer. Understanding these topics is essential for educating the public and medical professionals about potential risks associated with developmental exposure to BPA and other endocrine disruptors, the design of rigorously researched programs using both epidemiological and animal studies, and ultimately the development of a sound public health policy.

Comparative susceptibility to atrazine of three developmental stages of Rhinella arenarum and influence on metamorphosis: Non-monotonous acceleration of the time to climax and delayed tail resorption

Acute and subchronic toxicity of atrazine was evaluated in embryos (stage 4) and in premetamorphosis (stage 25) and prometamorphosis (stage 38–39) larvae of the common South American toad Rhinella arenarum (Anura: bufonidae). The influence of atrazine on the last stages of metamorphosis was also examined by exposing prometamorphosis larvae until completion of metamorphosis. Results obtained revealed that larvae in premetamorphosis are more sensitive than larvae in prometamorphosis and that these are, in turn, more sensitive than embryonic stages. Indeed, concentrations of atrazine as high as 30 mg/L had little effects on embryonic stages, the embryos surviving and developing in a similar manner as controls. LC50s of premetamorphosis larvae equaled 27.16, 7.03 and 2.32 mg/L of atrazine after 4, 14 and 21 days of exposure, respectively, compared to LC50s values of 18.27 and 14.43 mg/L after 14 and 21 days of exposure for larvae in prometamorphosis. In experiments with premetamorphosis larvae, the range of tested concentrations was extended to very low concentrations (down to 0.0001 mg/L) to examine whether recent findings of greater mortality at lower doses than at higher doses were also observed in R. arenarum but no such pattern was found. Exposure of prometamorphosis larvae to concentrations of atrazine of 10 mg/L and above widely prevented completion of metamorphosis and caused important mortality. Alternatively, whereas all animals eventually completed metamorphosis when exposed to concentrations of atrazine between 0.1 and 5 mg/L, the timings of metamorphosis were altered starting from 0.1 mg/L, the lowest concentration tested. Indeed, a significant decrease in the time needed for 50% of the larvae to reach the metamorphic climax (stage 42) was observed within this range of atrazine concentrations, the response presenting a U-shaped non-monotonic dose–response curve. Larvae exposed to these concentrations of atrazine also needed significantly more time for completing tail resorption, this effect being equivalent at all concentrations. Overall, the combination of these two different facets of atrazine influence on metamorphosis resulted in a significant acceleration of metamorphosis at 1 mg/L and a significant increase in the duration of metamorphosis at 5 mg/L, whereas no significant difference was observed with 0.1 mg/L.

Two-Generation Reproductive Toxicity Study of Dietary Bisphenol A in CD-1 (Swiss) Mice

Dietary bisphenol A (BPA) was evaluated in a mouse two-generation study at 0, 0.018, 0.18, 1.8, 30, 300, or 3500 ppm (0, 0.003, 0.03, 0.3, 5, 50, or 600 mg BPA/kg/day, 28 per sex per group). A concurrent positive control group of dietary 17beta-estradiol (0.5 ppm; 28 per sex) confirmed the sensitivity of CD-1 mice to an endogenous estrogen. There were no BPA-related effects on adult mating, fertility or gestational indices, ovarian primordial follicle counts, estrous cyclicity, precoital interval, offspring sex ratios or postnatal survival, sperm parameters or reproductive organ weights or histopathology (including the testes and prostate). Adult systemic effects: at 300 ppm, only centrilobular hepatocyte hypertrophy; at 3500 ppm, reduced body weight, increased kidney and liver weights, centrilobular hepatocyte hypertrophy, and renal nephropathy in males. At 3500 ppm, BPA also reduced F1/F2 weanling body weight, reduced weanling spleen and testes weights (with seminiferous tubule hypoplasia), slightly delayed preputial separation (PPS), and apparently increased the incidence of treatment-related, undescended testes only in weanlings, which did not result in adverse effects on adult reproductive structures or functions; this last finding is considered a developmental delay in the normal process of testes descent. It is likely that these transient effects were secondary to (and caused by) systemic toxicity. Gestational length was increased by 0.3 days in F1/F2 generations; the toxicological significance, if any, of this marginal difference is unknown. At lower doses (0.018-30 ppm), there were no treatment-related effects and no evidence of nonmonotonic dose-response curves for any parameter. The systemic no observable effect level (NOEL) was 30 ppm BPA (approximately 5 mg/kg/day); the reproductive/developmental NOEL was 300 ppm (approximately 50 mg/kg/day). Therefore, BPA is not considered a selective reproductive or developmental toxicant in mice.

Prenatal Lead Exposure in Mice: Age-Related, Sex-Specific Effects Observed

Children with low-level prenatal lead exposure may suffer reduced cognitive function, impaired motor ability, and visual and auditory processing problems. Other effects, such as accelerated age-related functional decline or delayed neurotoxicity, may become apparent in adulthood, though few studies have examined the long-term consequences of exposure. A novel animal model now reveals age-related, male-specific, and nonmonotonic dose–response effects associated with low-level prenatal lead exposure [EHP 116:355–361; Leasure et al.]. To model gestational lead exposure (GLE), one group of dams received tap water or drinking solutions containing low (27 ppm), moderate (55 ppm), or high (109 ppm) concentrations of lead beginning 2 weeks before mating and continuing until postnatal day 10. To measure postnatal lead exposure (PLE), another group of dams received tap water or water that contained low or moderate levels of lead from birth to weaning. The offspring of both groups were measured at birth and several times throughout the following year for weight and blood lead concentrations. Blood lead levels ranged from 10 μg/dL or less in low-exposure GLE offspring to 42 μg/dL in the high-exposure GLE group at post-natal day 10, and from 10 μg/dL in low-exposure PLE offspring to 27 μg/dL in the high-exposure PLE group at postnatal day 21. By postnatal day 30 for GLE offspring and postnatal day 60 for PLE offspring, blood lead levels were no different than in controls. At 1 year of age, male and female offspring were assessed for exploratory activity and interlimb balance and coordination. Because human developmental studies have demonstrated that males are at higher risk of deficits related to early lead exposure, additional testing of male offspring included measuring fore-brain and striatal levels of dopamine and its major metabolite, 3,4-dihydroxyphenylacetic acid. Lead exposure did not affect body weight of any PLE mice or of female GLE mice, but there was a significant inverse relationship between lead exposure and body weight for male low-exposure and high-exposure GLE mice (26% and 13% heavier than controls, respectively). Male GLE mice also exhibited significantly less exploratory activity, with a greater effect again seen in the low-exposure group. This group also had significantly poorer balance and coordination, lower forebrain levels of dopamine, and higher forebrain and striatal levels of 3,4-dihydroxyphenylacetic acid. Multiple endocrine or metabolic mechanisms that occurred during gestational development could explain the late-onset obesity observed at 1 year of age. The diminished exploratory activity may reflect an altered stress response arising from lead-related changes in the hypothalamic–pituitary–adrenal axis and dopaminergic systems. Changes in the forebrain may also have implications for attention deficit/hyperactivity disorder, as abnormalities in the prefrontal cortex appear to underlie the disorder. Multidisciplinary behavioral, biochemical, and molecular studies are needed to test and explain the reported novel findings, which highlight the need for realistic lifetime dose–response assessment of toxicants.

Hormesis [Biological Effects of Low Level Exposures (Belle)] and Dermatology

Hormesis, or biological effects of low level exposures (BELLE), is characterized by nonmonotonic dose response which is biphasic, displaying opposite effects at low and high dose. Its occurrence has been documented across a broad range of biological models and diverse type of exposure. Since hormesis appears to be a relatively common phenomenon in many areas, the objective of this review is to explore its occurrence related to dermatology and its public health and risk assessment implication. Hormesis appears to be a common phenomenon in in-vitro skin biology. However, in vivo data are lacking and the clinical relevance of hormesis has yet to be determined. Better understanding of this phenomenon will likely lead to different strategies for risk assessment process employed in the fields of dermatologic toxicology and pharmacology. We believe that hormesis is a common phenomenon and should be given detailed consideration to its concept and its risk assessment implications, and how these may be incorporated into the experimental and regulatory processes in dermatology. The skin, with its unique characteristics, its accessibility, and the availability of non-invasive bioengineering and DNA microarray technology, will be a good candidate to extend the biology of hormesis.

The importance of hormesis to public health

Hormesis is a specific type of nonmonotonic dose response whose occurrence has been documented across a broad range of biological models and diverse types of exposure. The effects that occur at various points along this curve can be interpreted as beneficial or detrimental, depending on the biological or ecologic context in which they occur. Because hormesis appears to be a relatively common phenomenon that has not yet been incorporated into regulatory practice, the objective of this commentary is to explore some of its more obvious public health and risk assessment implications, with particular reference to issues raised recently within this journal by other authors. Hormesis appears to be more common than doseresponse curves that are currently used in the risk assessment process. Although a number of mechanisms have been identified that explain many hormetic doseresponse relationships, better understanding of this phenomenon will likely lead to different strategies not only for the prevention and treatment of disease but also for the promotion of improved public health as it relates to both specific and more holistic health outcomes. We believe that ignoring hormesis is poor policy because it ignores knowledge that could be used to improve public health.

Non-monotonic dose–response relationship in steroid hormone receptor-mediated gene expression

Steroid hormone receptors are the targets of many environmental endocrine active chemicals (EACs) and synthetic drugs used in hormone therapy. While most of these chemical compounds have a unidirectional and monotonic effect, certain EACs can display non-monotonic dose-response behaviors and some synthetic drugs are selective endocrine modulators. Mechanisms underlying these complex endocrine behaviors have not been fully understood. By formulating an ordinary differential equation-based computational model, we investigated in this study the steady-state dose-response behavior of exogenous steroid ligands in an endogenous hormonal background under various parameter conditions. Our simulation revealed that non-monotonic dose-responses in gene expression can arise within the classical genomic framework of steroid signaling. Specifically, when the exogenous ligand is an agonist, a U-shaped dose-response appears as a result of the inherently nonlinear process of receptor homodimerization. This U-shaped dose-response curve can be further modulated by mixed-ligand heterodimers formed between endogenous ligand-bound and exogenous ligand-bound receptor monomers. When the heterodimer is transcriptionally inactive or repressive, the magnitude of U-shape increases; conversely, when the heterodimer is transcriptionally active, the magnitude of U-shape decreases. Additionally, we found that an inverted U-shaped dose-response can arise when the heterodimer is a strong transcription activator regardless of whether the exogenous ligand is an agonist or antagonist. Our work provides a novel mechanism for non-monotonic, particularly U-shaped, dose-response behaviors observed with certain steroid mimics, and may help not only understand how selective steroid receptor modulators work but also improve risk assessment for EACs.

Histopathological alterations, EROD activity, CYP1A protein and biliary metabolites in gilthead seabream Sparus aurata exposed to benzo(a)pyrene.

This study compared for seabream, Sparus aurata exposed to benzo(a)pyrene-B(a)P-, the response of molecular cytochrome P450 1A (CYP1A) and cellular histopathology biomarkers. Male gilthead seabream, Sparus aurata specimens were exposed for 20 days via water to a series of high B(a)P concentrations. CYP1A was assessed by measuring enzymatic activity (EROD) and CYP1A protein content, and cellular responses were evaluated by routine histopathological methods. In addition, biliary metabolites were measured in order to verify that B(a)P was absorbed and metabolised. Histological lesions, both in liver and gills, increased in parallel to B(a)P concentrations, with the majority of changes representing rather non-specific alterations. Hepatic EROD and CYP1A proteins data showed a concentration-dependent induction, while in the gills, EROD activity but not CYP1A proteins showed a non-monotonous dose response, with a maximum induction level at 200 microg B(a)P.L-1 and decreasing levels thereafter. The findings provide evidence that short-term, high dose exposure of fish can result in significant uptake and metabolism of the lipophilic B(a)P, and in pronounced pathological damage of absorptive epithelia and internal organs.

Effects of 17α‐ethinylestradiol and bisphenol a on vertebral development in the fathead minnow (Pimephales Promelas)

Growth, reproductive ability, and metabolic functions may be impaired by disruption of early endocrine patterning. Natural and synthetic estrogens detected in surface waters have been linked to reproductive endocrine signaling disruption in several species. The present study characterizes the nonreproductive morphological endpoint of vertebral anomalies in fish exposed to environmental estrogens. Estrogen is a proliferation-inducing compound in osteoblasts, regulating cartilage and bone deposition during development in vertebrates. The hypothesis for the present work is that xenobiotics with estrogenic activity adversely impact vertebral bone formation. Fathead minnows (Pimephales promelas) were exposed to 0.1 to 100 microg/L 17alpha-ethinylestradiol (EE2) and 0.1 to 1,000 microg/L bisphenol A (BPA) from egg stage (24 h postfertilization) to 25 to 26 d posthatch. Fish were measured for length and analyzed microscopically to determine degree of skeletal development (developmental score) and the occurrence of spinal abnormalities, including vertebral compression, bone fusion, and spinal curvatures. Fish length and developmental score were inversely related to vertebral malformations in exposed fish. Skeletal developmental was affected significantly in EE2-exposed fish: Vertebral malformations were observed in up to 62% of fish in a nonmonotonic dose-response. However, BPA did not significantly impair skeletal development or induce vertebral malformations. The bioassay results suggest vertebral bone development is a potential endpoint of endocrine disruption from potent estrogenic compounds in surface waters.

Hormesis and Its Place in Nonmonotonic Dose–Response Relationships: Some Scientific Reality Checks

ObjectiveThis analysis is a critical assessment of current hormesis literature. I discuss definitions, characterization, generalizability, mechanisms, absence of empirical data specific for hormesis hypothesis testing, and arguments that hormesis be the “default assumption” in risk assessment.Data sourcesHormesis, a biological phenomenon typically described as low-dose stimulation from substances producing higher-dose inhibition, has recently garnered interest in several quarters. The principal sources of published materials for this analysis are the writings of certain proponents of hormesis. Surprisingly few systematic critiques of current hormesis literature exist. Limits to the phenomenon’s appropriate role in risk assessment and health policy have been published.Data synthesisSerious gaps in scientific understanding remain: a stable definition; generalizability, especially for humans; a clear mechanistic basis; limitations in the presence of multiple toxic end points, target organs, and mechanisms. Absence of both arms-length, consensus-driven, scientific evaluations and empirical data from studies specifically designed for hormesis testing have limited its acceptance.ConclusionsDefinition, characterization, occurrence, and mechanistic rationale for hormesis will remain speculative, absent rigorous studies done specifically for hormesis testing. Any role for hormesis in current risk assessment and regulatory policies for toxics remains to be determined.