Validation Study Report: Performance assessment of the AR-CALUX® in vitro method: to support the development of an international test guideline for Androgen Receptor Transactivation Assays (ARTA) for the detection of compounds with (anti)androgenic potential

The JRC’s EU Reference Laboratory for alternatives to animal testing (EURL ECVAM) conducted a validation study of the AR-CALUX in vitro method. The method is applied to the detection of compounds with endocrine disrupting potential and more specifically (anti)androgen activity. The objectives of the study included assessing the reproducibility (within and between laboratories) and the relevance of the in vitro method. The participating laboratories included three test facilities from the European Union Network of Validation Laboratories for alternative methods (EU-NETVAL), being RISE, Covance, and Charles River, and, the test method submitter Biodetection Systems. The validation study report presents the results of the method performed on 46 test chemicals, evaluated for reproducibility within and between laboratories, variability within the measurements, and classification. A comparison of the classifications was made with publicly available ARTA classifications. It was concluded that this test method is reliable, has high reproducibility, low variability and merits proposal to OECD for the development of a test guideline. The statistical report and the final SOP are part of this report and can be found as separate files.JRC.F.3-Chemicals Safety and Alternative Method

The European Chemicals Bureau: an Overview of 15 Years Experience in EU Chemicals Legislation

From its creation in 1993, the European Chemicals Bureau (ECB) has played a vital role in the conception, development, implementation and monitoring of European Union (EU) legislation on chemicals and in contributing to the European Commission¿s participation in international chemicals programmes. The ECB has housed much of the European Commission¿s experience, capacity and historical memory in chemical risk assessment and safe chemical management. The contribution of ECB to the drafting, development and implementation of the REACH regulation has been an important one. The provision of scientific/technical expertise to the start-up phase of the newly born European Chemicals Agency (ECHA) has been essential for a swift and effective implementation of REACH. The ECB has contributed to that effort not only by selecting, recruiting and training ECHA staff but also by seconding part of its own key staff to the agency. And finally, during 2008 the ECB is completing the hand-over files and transmitting them to the ECHA, which is taking over responsibility for the operational implementation of EU legislation on chemicals.JRC.I-Institute for Health and Consumer Protection (Ispra

Entwicklung, Evaluation und Prävalidierung eines Immunotoxizitätstests

There are several ways how xenobiotics can influence the human immune system. Human whole blood was used to model the human immune system in vitro. The first publication of this thesis illustrates opportunities of a whole blood cytokine response based immunotoxicity test. Immunosuppressing compounds were shown to potently suppress cytokine responses to LPS in vitro, as compared to non-immunotoxic compounds. In the second publication this concept of a immunotoxicity test based on human whole blood cytokine response was optimised, standardised and prevalidated. Potency of compounds to modulate cytokine release by human blood monocytes and lymphocytes in vitro was determined by measuring the release of interleukin 1â and interleukin 4, respectively. It was shown that certain pharmaceutical agents exert an immunostimulating effect on monocytes, resulting in an increased release of IL-1â. SC4 (i.e. 4-fold stimulating concentration) values were determined to quantify the stimulating effect. The IC50 values of immunosuppressing compounds were determined and compared to in vivo peak plasma concentrations. Compounds that do not require metabolic activation and which are not cytotoxic, had IC50 values falling in the range of therapeutic plasma concentrations. Although most compounds were suppressing IL-4 release stronger, there were a few agents that suppressed IL-1â release more pronounced, indicating specificity for a certain immune function. The inter- and intra-laboratory reproducibility was good, and correlation with human and animal in vivo data showed that the results of the test were meaningful for the in vivo situation. A standard operating procedure (SOP) was developed, and can be found in Annex I of this thesis. The immunotoxicity test, as presented here, offers the possibility to screen compounds for immunotoxic properties against monocytes and Th2 lymphocytes of the immune system

Whole Blood Cytokine Response as a Measure of Immunotoxicity.

Abstract not availableJRC.I-Institute for Health and Consumer Protection (Ispra

Evaluation and Prevalidation of an Immunotoxicity Test Based on Human Whole-Blood Cytokine Release.

Abstract not availableJRC.I-Institute for Health and Consumer Protection (Ispra

A Generically Engineered Cell-Based System for Detecting Metabolism-Mediated Toxicity.

Abstract not availableJRC.I-Institute for Health and Consumer Protection (Ispra

Metabolism and Neurotoxicity: the Significance of Genetically Engineered Cell Lines and New Three-Dimensional Cell Cultures.

Abstract not availableJRC.I-Institute for Health and Consumer Protection (Ispra

Considerations in the development of in vitro toxicity testing methods intended for regulatory use

It has been demonstrated that the performance of in vitro toxicity testing methods can be seriously affected by variability in the essential components of the method, including the experimental protocol, the test items (chemicals), the test system (the biological models), and the analytical or measurement technique(s) used. Therefore these aspects need careful consideration during the development of in vitro methods if they are to be used in regulatory applications where compliance with quality systems is typically a prerequisite. Concerning the test system, implementation of the basic concepts of Good Cell Culture Practices (GCCP) is essential in the identification and characterisation of the biological model (test system part of the in vitro method). An adequate quality system, preferably Good Laboratory Practice (GLP) together with GCCP leads to overall Good In vitro Method Practices (GIVIM). Additional guidance for test developers will help to ensure that in vitro methods destined for regulatory use will prove to be robust, reliable and ultimately useful for supporting regulatory decisions.JRC.I.5-Systems Toxicolog

Promoting Reusable and Open Methods and Protocols (PRO-MaP): Draft recommendations to improve methodological clarity in life sciences publications

Detailed, accessible methods are essential for reproducibility, trust in science and scientific advancement; yet, many studies suggest that the reporting of methodological details in life sciences research publications is often incomplete. This may be due to a lack of incentives or reporting standards, or other cultural or educational factors. Promoting Reusable and Open Methods and Protocols (PRO-MaP) aims to increase and improve the reporting of detailed, reusable and open methods and reusable step-by-step protocols in the life sciences. This initiative began with a workshop convened by the EU Reference Laboratory for alternatives to animal testing (EURL ECVAM) at the European Commission’s Joint Research Centre, which included representatives from various stakeholder groups. Our draft recommendations outline actions that four stakeholder groups, researchers, research institutions and departments, publishers and editors, and funders, can take to achieve these goals. While some recommendations address study design and reporting guidelines, the primary focus is on capturing clear, accurate, methodological detail, e.g. with re-usable step-by-step protocols. We welcome feedback on these draft recommendations from members of each stakeholder group, including experts in improving methodological reporting. The final recommendations will be released after incorporating feedback received during consultation sessions. Once the recommendations are finalized, we hope that organizations and individuals from each stakeholder group will join us in collaboratively working to improve the reporting of detailed methods and reusable step-by-step protocols in the life sciences