Erik Ropstad

Science and policy on endocrine disrupters must not be mixed: a reply to a “common sense” intervention by toxicology journal editors

The “common sense” intervention by toxicology journal editors regarding proposed European Union endocrine disrupter regulations ignores scientific evidence and well-established principles of chemical risk assessment. In this commentary, endocrine disrupter experts express their concerns about a recently published, and is in our considered opinion inaccurate and factually incorrect, editorial that has appeared in several journals in toxicology. Some of the shortcomings of the editorial are discussed in detail. We call for a better founded scientific debate which may help to overcome a polarisation of views detrimental to reaching a consensus about scientific foundations for endocrine disrupter regulation in the EU.

Reproductive and Developmental Toxicity of Phthalates

The purposes of this review are to (1) evaluate human and experimental evidence for adverse effects on reproduction and development in humans, produced by exposure to phthalates, and (2) identify knowledge gaps as for future studies. The widespread use of phthalates in consumer products leads to ubiquitous and constant exposure of humans to these chemicals. Phthalates were postulated to produce endocrine-disrupting effects in rodents, where fetal exposure to these compounds was found to induce developmental and reproductive toxicity. The adverse effects observed in rodent models raised concerns as to whether exposure to phthalates represents a potential health risk to humans. At present, di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP), and butyl benzyl phthalate (BBP) have been demonstrated to produce reproductive and developmental toxicity; thus, this review focuses on these chemicals. For the general population, DEHP exposure is predominantly via food. The average concentrations of phthalates are highest in children and decrease with age. At present, DEHP exposures in the general population appear to be close to the tolerable daily intake (TDI), suggesting that at least some individuals exceed the TDI. In addition, specific high-risk groups exist with internal levels that are several orders of magnitude above average. Urinary metabolites used as biomarkers for the internal levels provide additional means to determine more specifically phthalate exposure levels in both general and high-risk populations. However, exposure data are not consistent and there are indications that secondary metabolites may be more accurate indicators of the internal exposure compared to primary metabolites. The present human toxicity data are not sufficient for evaluating the occurrence of reproductive effects following phthalate exposure in humans, based on existing relevant animal data. This is especially the case for data on female reproductive toxicity, which are scarce. Therefore, future research needs to focus on developmental and reproductive endpoints in humans. It should be noted that phthalates occur in mixtures but most toxicological information is based on single compounds. Thus, it is concluded that it is important to improve the knowledge of toxic interactions among the different chemicals and to develop measures for combined exposure to various groups of phthalates.

The role of parasites in the dynamics of a reindeer population

Even though theoretical models show that parasites may regulate host population densities, few empirical studies have given support to this hypothesis. We present experimental and observational evidence for a host–parasite interaction where the parasite has sufficient impact on host population dynamics for regulation to occur. During a six year study of the Svalbard reindeer and its parasitic gastrointestinal nematode Ostertagia gruehneri we found that anthelminthic treatment in April–May increased the probability of a reindeer having a calf in the next year, compared with untreated controls. However, treatment did not influence the over–winter survival of the reindeer. The annual variation in the degree to which parasites depressed fecundity was positively related to the abundance of O. gruehneri infection the previous October, which in turn was related to host density two years earlier. In addition to the treatment effect, there was a strong negative effect of winter precipitation on the probability of female reindeer having a calf. A simple matrix model was parameterized using estimates from our experimental and observational data. This model shows that the parasite–mediated effect on fecundity was sufficient to regulate reindeer densities around observed host densities.

Organochlorines Affect the Major Androgenic Hormone, Testosterone, in Male Polar Bears (Ursus Maritimus) at Svalbard

Normal sexual development and subsequent reproductive function are dependent on appropriate testosterone production and action. The regulation of steroid hormones, including androgens, can be influenced by both biological and environmental factors, including environmental chemicals. Concentrations of organochlorines are considerably greater in Svalbard polar bears than in polar bears from other regions. Between 1995 and 1998, samples were collected from 121 male polar bears (Ursus maritimus) from the Svalbard area. In this study, testosterone concentration variations were described for male polar bears during different seasons and for all age groups. To study possible relationships between plasma testosterone concentrations and biological factors, such as age, axial girth, and extractable plasma fat, and organochlorine contaminants including hexachlorocyclohexanes, hexachlorobenzene, chlordanes, p,p′–DDE, and 16 individual polychlorinated biphenyl (PCB) congeners, identical statistical analyses were performed on the total population and a subsample of reproductively active adults. Of the biological factors, axial girth showed a significant positive relationship and percentage extractable fat and a significant negative relationship with the testosterone concentrations. Both the Σpesticides and ΣPCBs made significant negative contributions to the variation of the plasma testosterone concentration. The continuous presence of high concentrations of organochlorines in male polar bears throughout their life could possibly aggravate any reproductive toxicity that may have occurred during fetal and early postnatal development.

Endocrine disrupting effects of zearalenone, alpha- and beta-zearalenol at the level of nuclear receptor binding and steroidogenesis.

The mycotoxin zearalenone (ZEN) is a secondary metabolite of fungi which is produced by certain species of the genus Fusarium and can occur in cereals and other plant products. Reporter gene assays incorporating natural steroid receptors and the H295R steroidogenesis assay have been implemented to assess the endocrine disrupting activity of ZEN and its metabolites α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL). α-ZOL exhibited the strongest estrogenic potency (EC(50) 0.022±0.001 nM), slightly less potent than 17-β estradiol (EC(50) 0.015±0.002 nM). ZEN was ~70 times less potent than α-ZOL and twice as potent as β-ZOL. Binding of progesterone to the progestagen receptor was shown to be synergistically increased in the presence of ZEN, α-ZOL or β-ZOL. ZEN, α-ZOL or β-ZOL increased production of progesterone, estradiol, testosterone and cortisol hormones in the H295R steroidogenesis assay, with peak productions at 10 μM. At 100 μM, cell viability decreased and levels of hormones were significantly reduced except for progesterone. β-ZOL increased estradiol concentrations more than α-ZOL or ZEN, with a maximum effect at 10 μM, with β-ZOL (562±59 pg/ml)>α-ZOL (494±60 pg/ml)>ZEN (375±43 pg/ml). The results indicate that ZEN and its metabolites can act as potential endocrine disruptors at the level of nuclear receptor signalling and by altering hormone production.

Organochlorines Affect the Steroid Hormone Cortisol in Free-Ranging Polar Bears (Ursus maritimus) at Svalbard, Norway

Since the polar bear (Ursus maritimus) is among the most highly organochlorine-contaminated species of the Arctic mammals, there is growing concern that in addition to the natural stressors in the polar bears environment, several organochlorines (OCs) may be able to change basic endocrine pathways. Alterations in the hypothalamic–pituitary–adrenal (HPA) axis may affect plasma cortisol concentrations and inhibit physiological processes involved in the maintenance of homeostasis in a way that may endanger the animals health. Between 1995 and 1998, samples were collected from 121 male and 130 female free-ranging polar bears from the Svalbard area. The aim of the study was to investigate relationships between plasma cortisol concentrations, biological factors, and OCs. The variation in plasma cortisol concentrations was determined for the total sample. Axillary girth and body mass together with their interactions explained more than 50% of the variation in the plasma cortisol concentration. The sum of pesticides (Σpesticides) combined with the sum of polychlorinated biphenyls (ΣPCBs) and their interactions explained over 25% of the variation in the cortisol concentration. Although Σpesticides contributed negatively and ΣPCBs contributed positively to the variation in the plasma cortisol, the over-all contribution of the OCs to the plasma cortisol variation was negative. Despite the complexity on stress responses and the interactions with environmental factors, this study demonstrated that high concentrations of OCs in polar bears might alter plasma cortisol concentrations.

In Vitro Assay Shows That PCB Metabolites Completely Saturate Thyroid Hormone Transport Capacity in Blood of Wild Polar Bears (Ursus maritimus)

Persistent chemicals accumulate in the arctic environment due to their chemical reactivity and physicochemical properties and polychlorinated biphenyls (PCBs) are the most concentrated pollutant class in polar bears (Ursus maritimus). Metabolism of PCB and polybrominated biphenyl ether (PBDE) flame-retardants alter their toxicological properties and these metabolites are known to interfere with the binding of thyroid hormone (TH) to transthyretin (TTR) in rodents and humans. In polar bear plasma samples no binding of [125I]-T(4) to TTR was observed after incubation and PAGE separation. Incubation of the plasma samples with [14C]-4-OH-CB107, a compound with a higher binding affinity to TTR than the endogenous ligand T(4) resulted in competitive binding as proven by the appearance of a radio labeled TTR peak in the gel. Plasma incubation with T(4) up to 1 mM, a concentration that is not physiologically relevant anymore did not result in any visible competition. These results give evidence that the binding sites on TTR for T(4) in wild living polar bears are completely saturated. Such saturation of binding sites can explain observed lowered levels of THs and could lead to contaminant transport into the developing fetus.

Valproate inhibits the conversion of testosterone to estradiol and acts as an apoptotic agent in growing porcine ovarian follicular cells.

Summary:  Purpose: Long‐term valproate (VPA) treatment has been associated with hyperandrogenism and polycystic ovaries in women with epilepsy. The exact mechanisms of action of the drug on sex steroid hormone function are still unsettled. The aim of the present study was to investigate the action of VPA on basal and gonadotropin‐stimulated steroid secretion in porcine ovarian follicular cells and to measure the conversion of testosterone to estradiol. Second, the action of VPA on proliferation and apoptosis of follicular cells was investigated.

Congruent responses to weather variability in high arctic herbivores

Assessing the role of weather in the dynamics of wildlife populations is a pressing task in the face of rapid environmental change. Rodents and ruminants are abundant herbivore species in most Arctic ecosystems, many of which are experiencing particularly rapid climate change. Their different life-history characteristics, with the exception of their trophic position, suggest that they should show different responses to environmental variation. Here we show that the only mammalian herbivores on the Arctic islands of Svalbard, reindeer (Rangifer tarandus) and sibling voles (Microtus levis), exhibit strong synchrony in population parameters. This synchrony is due to rain-on-snow events that cause ground ice and demonstrates that climate impacts can be similarly integrated and expressed in species with highly contrasting life histories. The finding suggests that responses of wildlife populations to climate variability and change might be more consistent in Polar regions than elsewhere owing to the strength of the climate impact and the simplicity of the ecosystem.

Organohalogen contaminants and reproductive hormones in incubating glaucous gulls (Larus hyperboreus) from the Norwegian Arctic

Organohalogen contaminants detected globally in avian wildlife, including populations from the Arctic, have been related to various reproductive hormone potencies, and altered hormonal balance and functions. Besides legacy organochlorine (OC) substances, that is, polychlorinated biphenyls (PCBs) and OC pesticides and by-products, endocrine-disruptive properties have been defined for chemicals of new and emerging environmental concern, such as polybrominated diphenyl ethers (PBDEs) and metabolically derived products like methylsulfonyl (MeSO2)- and hydroxyl (OH)-PCBs. We investigated the relationships between plasma concentrations of selected legacy OCs, PBDEs, and MeSO2- and OH-PCB metabolites and the circulating reproductive hormones testosterone (T), 17beta-estradiol (E2), and progesterone (P4) in incubating male and female glaucous gulls (Larus hyperboreus) from the Norwegian Arctic. Principal component and regression analyses demonstrated that P4 levels in male glaucous gulls were associated positively with variations of sum (Y) PCB, dichloro-diphenyl-trichloroethane (sigmaDDT), chlordane (sigmaCHL), and sigmaPBDE concentrations, which were the most recalcitrant organohalogens determined in glaucous gulls. No such relationship was found for female glaucous gulls as well as between concentrations of any of the selected organohalogens and levels of T for both sexes. The E2 was not detected in any plasma samples. Present results were highly suggestive that exposure to high organohalogen concentrations in glaucous gulls, particularly the most persistent compound classes, may have the potential to interfere with steroidogenesis and impinge on circulating P4 homeostasis. Because significant effects were found in males exclusively, it cannot be completely ruled out that male glaucous gulls are more sensitive than females to organohalogen-mediated alteration of P4 synthesis and breakdown.