The OECD validation program of the H295R steroidogenesis assay: Phase 3. Final inter-laboratory validation study

Hecker, Markus; Hollert, Henner; Cooper, Ralph L.; Vinggaard, Anne Marie; Akahori, Yumi; Murphy, Margaret B.; Nellemann, Christine; Higley, Eric; Newsted, John L.; Laskey, John W.; Buckalew, Angela R.; Grund, Stefanie; Maletz, Sibylle; Giesy, John P.; Timm, Gary

doi:10.1007/s11356-010-0396-x

Cited by 83 publications

(63 citation statements)

References 42 publications

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“…PBO was tested for (anti-)estrogenic and (anti-)androgenic effect in the OECD-standardized H295R steroidogenesis in vitro assay (Hecker et al 2011). PBO tested negative for E2 production with 100 µM as the highest tested concentration and the LOEC for testosterone production was 10 mg/l (30 µM).…”

Section: In Vitro Studiesmentioning

confidence: 99%

“…In vitro studies have demonstrated that PBO affects steroidogenesis (Hecker et al 2011). Non EATS-modalities were also demonstrated in vitro as PBO showed immunotoxicity (Battaglia et al 2010) and a negative effect on the release of corticotropin (Luini and Axelrod 1985).…”

Section: Piperonyl Butoxidementioning

confidence: 99%

“…Hecker et al (2011) used the H295R Steroidogenesis Assay to demonstrate that PBO has a negative effect on the production of testosterone but seemingly no effect on the production of 17beta-estradiol. This method is equal to the OECD TG 456.…”

Section: Piperonyl Butoxidementioning

confidence: 99%

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Suspected endocrine disrupting substances

Publicering¹

2013

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Section: In Vitro Studiesmentioning

confidence: 99%

Section: Piperonyl Butoxidementioning

confidence: 99%

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Suspected endocrine disrupting substances

Publicering¹

2013

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“…The effects of the single pesticides and the two pesticide mixtures on the production of progesterone, testosterone, and estradiol were tested in the NCI-H295R human adrenocortical carcinoma cell line (ATCC no. CRL-2128, LGC Standards, Boras, Sweden) as previously described (Hecker et al, 2011). In brief cells were seeded in 24-well culture plates (Costar3524,Corning,NY, USA) with DMEM/F12 medium (Gibco, Paisley, UK) supplemented with 2.0% Nu-serum (BD Sciences Denmark) along with 1% ITS+ premix (containing6.25 μg/ml insulin, 6.25 μg/ml transferrin,6.25 ng/ml selenium,1.25 mg/ml BSA and 5.35 μg/ml linoleic acid; BD Sciences Denmark) and incubated for 24 h at37°C in a humidified atmosphere of 5% CO2/air.…”

Section: H295r Assaymentioning

confidence: 99%

In vitro–in vivo correlations for endocrine activity of a mixture of 5 currently used pesticides

et al. 2013

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Two pesticide mixtures were investigated for potential endocrine activity. Mix 3 consisted of bitertanol, propiconazole, and cypermethrin, and Mix 5 included malathion and terbuthylazine in addition to the three pesticides in Mix 3. All five single pesticides and the two mixtures were investigated for their ability to affect steroidogenesis in vitro in H295R cells. The pesticides alone and both mixtures affected steroidogenesis with both mixtures causing increase in progesterone and decrease in testosterone. For Mix 5 an increase in estradiol was seen as well, indicating increased aromatase activity. The two mixtures were also investigated in pregnant rats dosed from gestational day 7 to 21, followed by examination of dams and fetuses. Decreased estradiol and reduced placental testosterone were seen in dams exposed to Mix 5. Also a significant increase in aromatase mRNA-levels in female adrenal glands was found for Mix5. However, either of the two mixtures showed any effects on fetal hormone levels in plasma or testis, or on anogenital distance. Overall, potential aromatase induction was found for Mix 5 both in vitro and in vivo, but not for Mix 3, an effect likely owed to terbuthylazine in Mix 5. However, the hormonal responses in vitro were only partly reflected in vivo, probably due to some toxicokinetic issues, as the pesticide levels in the amniotic fluid also were found to be negatively affected by the number of compounds present in the mixtures. Nonetheless, the H295R assay gives hints on conceivable interference with steroidogenesis, thus generating hypotheses on in vivo effects.

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“…These assays serve as first-tier screens to identify the potential EDC activity and suggested MOA of man-made chemicals detected in the environment (Hecker et al, 2011). Although the recombinant yeast assay (using hAR) has not been included in the battery of first-tier tests suggested by the USEPA, the YAS has been recognised as an assay that it is a relatively inexpensive screening tool with a standardised protocol (Sohoni and Sumpter, 1998;De Jager et al, 2011;Kolle et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

The potential anti-androgenic effect of agricultural pesticides used in the Western Cape: In vitro investigation of mixture effects

Archer¹,

Wyk²

2015

WSA

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Although it is known that environmental chemicals can affect the oestrogenic system, far less attention has been paid to chemicals interacting with the androgen receptor (AR). Pesticides, particularly fungicides, have been shown to competitively bind or affect expression of the AR in an inhibiting manner. Few studies have addressed anti-androgenic effects of agrochemical use in South Africa. The aim of this study was to screen for the ability of commonly-used pesticides (mostly fungicides) in Western Cape agricultural areas to alter the binding of an androgen (DHT) to the human AR (hAR) using a recombinant yeast androgen screen (YAS), and also to test the additivity mixture interaction hypothesis when commonly-used pesticides with similar modes of action (MOAs) are exposed in mixture. Fungicides vinclozolin, folpet, procymidone, dimethomorph, fenarimol, mancozeb, and the insecticide chlorpyrifos, all independently antagonised the binding of the androgen dihydrotestosterone (DHT) to the AR in a dose-dependent manner. The fungicide mancozeb was found to be the most potent anti-androgen in the assay. Binary, equimolar mixtures of the pesticides also antagonised the binding of DHT to the AR, but at lower IC 50 concentration potencies relative to their individual counterparts. The mixtures of the majority of the selected pesticides did not conform to the expected additive mixture interaction. Only the mixture between dimethomorph and mancozeb showed an additive mixture response at IC 50 concentrations, and, therefore, revealed a more severe AR antagonistic effect compared to their individual counterparts. This study confirmed that pesticides regularly used in agriculture inhibit the binding of androgens to the AR, but when in mixture do not always conform to the predictive addition mixture response model. Also, high relative potencies of individual chemicals in the assay were suppressed when combined with less potent chemicals, showing that the potent chemicals may not be granted access to bind with the AR when exposed in mixture.

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The OECD validation program of the H295R steroidogenesis assay: Phase 3. Final inter-laboratory validation study

Abstract: Based on the results obtained during this validation study and the accordingly revised test protocols, an OECD draft test guideline was developed and submitted to the OECD working group of the national coordinators of the test guidelines program (WNT) for comments in December 2009.

Cited by 83 publications

References 42 publications

Suspected endocrine disrupting substances

Suspected endocrine disrupting substances

In vitro–in vivo correlations for endocrine activity of a mixture of 5 currently used pesticides

The potential anti-androgenic effect of agricultural pesticides used in the Western Cape: In vitro investigation of mixture effects

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