Modified Xenopus laevis approach (R-FETAX) as an alternative test for the evaluation of foetal valproate spectrum disorder

Battistoni, Maria; Bacchetta, Renato; Renzo, Francesca Di; Metruccio, Francesca; Moretto, Angelo; Menegola, Elena

doi:10.1016/j.reprotox.2021.12.005

Cited by 11 publications

(7 citation statements)

References 69 publications

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“…In parallel, to evaluate embryotoxicity, the effects of extracts were tested in Xenopus laevis embryos. Xenopus laevis embryos were exposed during the whole R-FETAX test period (from midblastula to tadpole, according to [61]) to the soluble extracted fraction diluted 1:10 in maintaining solution (see details in Supplementary Materials). At the end of the test, tadpoles were morphologically observed under a dissecting microscope (Leica).…”

Section: Filter-based Toxicological Assaysmentioning

confidence: 99%

On the Redox-Activity and Health-Effects of Atmospheric Primary and Secondary Aerosol: Phenomenology

et al. 2022

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The RHAPS (Redox-Activity And Health-Effects Of Atmospheric Primary And Secondary Aerosol) project was launched in 2019 with the major objective of identifying specific properties of the fine atmospheric aerosol from combustion sources that are responsible for toxicological effects and can be used as new metrics for health-related outdoor pollution studies. In this paper, we present the overall methodology of RHAPS and introduce the phenomenology and the first data observed. A comprehensive physico-chemical aerosol characterization has been achieved by means of high-time resolution measurements (e.g., number size distributions, refractory chemical components, elemental composition) and low-time resolution analyses (e.g., oxidative potential, toxicological assays, chemical composition). Preliminary results indicate that, at the real atmospheric conditions observed (i.e., daily PM1 from less than 4 to more than 50 μg m−3), high/low mass concentrations of PM1, as well as black carbon (BC) and water soluble Oxidative Potential (WSOP,) do not necessarily translate into high/low toxicity. Notably, these findings were observed during a variety of atmospheric conditions and aerosol properties and with different toxicological assessments. Findings suggest a higher complexity in the relations observed between atmospheric aerosol and toxicological endpoints that go beyond the currently used PM1 metrics. Finally, we provide an outlook to companion papers where data will be analyzed in more detail, with the focus on source apportionment of PM1 and the role of source emissions on aerosol toxicity, the OP as a predictive variable for PM1 toxicity, and the related role of SOA possessing redox-active capacity, exposure-response relationships for PM1, and air quality models to forecast PM1 toxicity.

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Section: Filter-based Toxicological Assaysmentioning

confidence: 99%

On the Redox-Activity and Health-Effects of Atmospheric Primary and Secondary Aerosol: Phenomenology

et al. 2022

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“…Amphibian X. laevis adults (Nasco, USA) were maintained in an automatic breeding system (Tecnoplus, Techniplast, Italy) under controlled water conditions (T = 20 ± 2 °C; pH = 7.5 ± 0.5; conductivity = 1000 ± 100 μS), 12-h light/ dark cycle (light from 7:00 AM to 7:00 PM) and fed with a semisynthetic diet twice a week (XE40 by Mucedola; Settimo Milanese, Italy). Embryos were obtained as described by Battistoni et al (2022) without adult human chorionic gonadotropin injection. The collected embryos were cleaned by gentle swirling in a 2.25% L-cysteine solution with an arranged pH of 8.0 and rinsed several times in FETAX solution, whose composition was 625 mg/L NaCl, 96 mg/L NaHCO 3 , 30 mg/L KCl, 15 mg/L CaCl 2 , 60 mg/L CaSO 4 •2H 2 O, and 70 mg/L MgSO 4 .…”

Section: R-fetaxmentioning

confidence: 99%

“…At day 6, the deglutition test was performed as previously described by Battistoni et al (2022), with some modifications: larvae were maintained for 2 h at 23 ± 0.5 °C in FETAX solution containing 25 μg/mL red polystyrene microparticles (1 µm diameter, Sigma). Larvae, overdosed at 4 °C with anesthetic (0.02% MS222, dissolved in FETAX solution), were evaluated under a dissecting microscope (Leica) for gross morphology and presence of microplastics in the intestine and photographed.…”

Section: Deglutition Test and Gross Morphological Evaluationmentioning

confidence: 99%

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Predictive assays for craniofacial malformations: evaluation in Xenopus laevis embryos exposed to triadimefon

et al. 2022

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Craniofacial defects are one of the most frequent abnormalities at birth, but their experimental evaluation in animal models requires complex procedures. The aim of the present work is the comparison of different methodologies to identify dose- and stage-related craniofacial malformations in Xenopuslaevis assay (R-FETAX, where the full cartilage evaluation, including flat mount technique, is the gold standard for skeletal defect detection). Different methods (external morphological evaluation of fresh samples, deglutition test, whole mount cartilage evaluation and Meckel–palatoquadrate angle measurements) were applied. Triadimefon (FON) was selected as the causative molecule as it is known to induce craniofacial defects in different animal models, including the amphibian X.laevis.FON exposure (0–31.25 μM) was scheduled to cover the whole 6-day test (from gastrula to free swimming tadpole stage) or each crucial developmental phases: gastrula, neurula, early morphogenesis, late morphogenesis, tadpole. Dose-dependent effects (fusions among craniofacial cartilages) were evident for groups exposed during the morphogenetic periods (neurula, early morphogenesis, late morphogenesis); gastrula was insensitive to the tested concentrations, tadpole group showed malformations only at 31.25 μM. The overall NOAEL was set at 3.9 μM. Results were evaluated applying benchmark dose (BMD) approach. The comparison of relative potencies from different methods showed deglutition as the only assay comparable with the gold standard (cartilage full evaluation).In conclusion, we suggest deglutition test as a reliable method for a rapid screening of craniofacial abnormalities in the alternative model X.laevis. This is a rapid, inexpensive and vital test allowing to preserve samples for the application of further morphological or molecular investigations.

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“…The concamerated heart is visible ventrally ( =). A'' Unexposed control embryo showing the same morphological characteristics described in (A-A' Maele-Fabry et al 1995), VPA 0-31.25-62.5-125-250-375-500-750 µM (Metruccio et al 2020;Battistoni et al 2022). Mixture effect evaluation was performed following the fix and moving design.…”

Section: Selection Of Compound Concentrationsmentioning

confidence: 99%

Moderate alcohol consumption during pregnancy increases potency of two different drugs (the antifungal fluconazole and the antiepileptic valproate) in inducing craniofacial defects: prediction by the in vitro rat whole embryo culture

et al. 2022

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The prenatal exposure to ethanol (Eth), fluconazole (FLUCO) and sodium valproate (VPA) is related to effects on development, producing characteristic syndromic pictures. Among embryotoxic effects described for the three molecules, the alteration on craniofacial morphogenesis is a common feature in humans and animal models, including rodent embryos developed in vitro. The aim of the present work is to evaluate the developmental effects of low Eth serum concentration (17 mM, corresponding to the legal limit to drive in UK, USA, Canada, and many other countries) in mixture with increasing realistic concentrations of the antifungal drug FLUCO (62.5–500 µM) or with increasing realistic concentrations of the antiepileptic drug VPA (31.25–250 µM). Groups exposed to Eth alone (17–127.5 mM), FLUCO alone (62.5–500 µM) or VPA alone (31.25–750 µM) were also included. The chosen alternative animal model was the post-implantation rat whole embryo culture (WEC). E9.5 embryos were exposed in vitro to the test molecules during the whole test period (48 h, corresponding to the developmental stages characteristics of any vertebrate, for human embryos post-fertilization days 23–31). Data were statistically analyzed and processed for modelling applying the benchmark dose (BMD) and relative potency factor (RPF) approaches. Concentration-related effects on facial outcomes were observed in all experimental groups, with a significant enhancement in the groups co-exposed with Eth in comparison to the single exposures. Data obtained by the present work suggest an additional alert for the assumption of even low levels of alcohol in pregnant women during FLUCO or VPA therapy.

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Modified Xenopus laevis approach (R-FETAX) as an alternative test for the evaluation of foetal valproate spectrum disorder

Cited by 11 publications

References 69 publications

On the Redox-Activity and Health-Effects of Atmospheric Primary and Secondary Aerosol: Phenomenology

On the Redox-Activity and Health-Effects of Atmospheric Primary and Secondary Aerosol: Phenomenology

Predictive assays for craniofacial malformations: evaluation in Xenopus laevis embryos exposed to triadimefon

Moderate alcohol consumption during pregnancy increases potency of two different drugs (the antifungal fluconazole and the antiepileptic valproate) in inducing craniofacial defects: prediction by the in vitro rat whole embryo culture

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