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ZusammenfassungÜberzählige Versuchstiere lassen sich auch bei bester Versuchsplanung und sorgfältigstem Zuchtmanagement nicht vermeiden, da bei der Zucht von Versuchstieren regelmäßig Tiere entstehen, die aufgrund verschiedenster Kriterien im geplanten Tierversuch, für den sie ursprünglich gezüchtet wurden, nicht eingesetzt werden können. Insbesondere bei genetisch veränderten Tieren scheidet eine Vermittlung oder Weiterverwendung für andere Versuchszwecke oder als Futtertier regelmäßig aus. Viele Einrichtungen können diese überzähligen Tiere aufgrund fehlender Haltungskapazitäten auch nicht bis an ihr natürliches Lebensende halten. In der Praxis stellt sich daher die ethisch und rechtlich viel diskutierte Frage, ob überzählige Versuchstiere unter Annahme eines vernünftigen Grundes nach 1 S. 2 TierSchG getötet werden dürfen. Im Kern geht es dabei um die im Einzelfall stets vorzunehmende Abwägung des Tierschutzes gem. Art. 20a GG und der Wissenschaftsfreiheit gem. Art. 5 Abs. 3 GG. Der vorliegende Beitrag möchte insbesondere untersuchen, ob eine Verfütterung von überzähligen Versuchstieren und die Unmöglichkeit einer artgerechten Haltung unter Einbeziehung aktueller Rechtsprechung für sich genommen ausreichen können, um eine Tötung von überzähligen Versuchstieren aus vernünftigem Grund zu rechtfertigen.

ZusammenfassungDas Ziel der EU-Versuchstierrichtlinie 2010/63/EU ist es, einheitliche rechtliche Rahmenbedingungen für Tierversuche in allen EU-Mitgliedstaaten zu schaffen. Aufgrund eigenständiger mitgliedstaatlicher Umsetzung der EU-Versuchstierrichtlinie in das jeweilige nationale Versuchstierrecht können nationale Regelungen durchaus ein unterschiedlich hohes Tierschutzniveau aufweisen. Deutlich zeigt sich dies bei der Tötung von überzähligen Versuchstieren, die regelmäßig bei der notwendigen Zucht von Versuchstieren entstehen. Gem. 1 S. 2 TierSchG muss nach deutschem Tierschutzrecht bei jeder Tötung eines Tieres ein vernünftiger Grund vorliegen. Dieses Erfordernis bezieht sich uneingeschränkt auch auf Versuchstiere und zeigt sich in seiner konkreten Ausgestaltung als nationale Besonderheit des deutschen Versuchstierrechts im Vergleich zu anderen Mitgliedstaaten. Fehlt ein vernünftiger Grund im Rahmen der Tötung eines Tieres, kann dies auch strafrechtliche Konsequenzen gem. 17 Nr. 1 TierSchG nach sich ziehen. Mangels gefestigter Rechtsprechung im Versuchstierbereich muss bei der rechtlich gebotenen Auslegung zur Bestimmung des unbestimmten Rechtsbegriffs des vernünftigen Grundes die Rechtsprechung aus anderen Bereiches des Tierschutzrechts herangezogen und auf das Versuchstierrecht übertragen werden. Unter Heranziehung der aktuellsten Rechtsprechung des Bundesverwaltungsgerichts zur Tötung von männlichen Küken zeigt sich, wie schwierig sich eine solche Übertragung in der rechtswissenschaftlichen Praxis gestaltet. Eine mögliche Übertragung von Wertungen aus dem Nutztierbereich muss dabei aufgrund der Komplexität der versuchstierrechtlichen Materie stets den konkreten Einzelfall berücksichtigen und kann keinesfalls pauschal erfolgen.

Chapter 37 was previously published non-open access. It has now been changed to open access under a CC BY 4.0 license and the copyright holder updated to ‘The Author(s)’. The book has also been updated with this change.

Recent publications describe the development of in vitro models of human development, for which applications in developmental toxicity testing can be envisaged. To date, these regulatory assessments have exclusively been performed in animal studies, the relevance of which to adverse reactions in humans may be questioned. Recently developed cell culture-based models of embryo-fetal development, however, do not yet exhibit sufficient levels of standardisation and reproducibility. Here, the advantages and shortcomings of both in vivo and in vitro developmental toxicity testing are addressed, as well as the possibility of integrated testing strategies as a viable option in the near future.

Many widely used chemicals result in ubiquitous human exposure from multiple sources, including diet. Legislation mainly deals with the toxicological evaluation of single substances owing to a methodological and conceptual lack of alternatives, and does so within defined silos subject to over 40 distinct regulations in the EU alone. Furthermore, much of the research and many of the initiatives concerned with the assessment and evaluation of chemical mixtures and their potential effects on human health rely on retrospective analysis. Here we propose an approach for the prospective identification, assessment and regulation of mixtures relevant to human health. We address two distinct aspects of toxicology—which chemicals actually do occur together, and how potential mixture-related health hazards can be predicted—with an adapted concept of the exposome and large-scale hazard screens. The proactive use of the likelihood of co-exposure, together with the new approach of methods-based testing, may be a timely and feasible way of identifying those substances and mixtures where hazards may have been overlooked and regulatory action is needed. Ideally, we would generate co-exposure patterns for specific consumer groups, depending on lifestyle and dietary habits, to assess the specific risk of identified mixtures. Continuous and chronic exposure to mixtures of multiple chemicals, particularly from food, medicine and cleaning products, are of concern. A proactive combinatorial approach to identify, assess and regulate the likelihood of exposure and potential health effects is presented.

Preregistration of studies is a recognized tool in clinical research to improve the quality and reporting of all gained results. In preclinical research, preregistration could boost the translation of published results into clinical breakthroughs. When studies rely on animal testing or form the basis of clinical trials, maximizing the validity and reliability of research outcomes becomes in addition an ethical obligation. Nevertheless, the implementation of preregistration in animal research is still slow. However, research institutions, funders, and publishers start valuing preregistration, and thereby level the way for its broader acceptance in the future. A total of 3 public registries, the OSF registry, preclinicaltrials.eu, and animalstudyregistry.org already encourage the preregistration of research involving animals. Here, they jointly declare common standards to make preregistration a valuable tool for better science. Registries should meet the following criteria: public accessibility, transparency in their financial sources, tracking of changes, and warranty and sustainability of data. Furthermore, registration templates should cover a minimum set of mandatory information and studies have to be uniquely identifiable. Finally, preregistered studies should be linked to any published outcome. To ensure that preregistration becomes a powerful instrument, publishers, funders, and institutions should refer to registries that fulfill these minimum standards.

Attempts to replicate published studies have too often failed, and promising preclinical results could not be translated into clinical breakthroughs. Major causes for this lack of reproducibility are practical reasons such as selective reporting, poor study design, or insufficient method description. However, a rigid incentive system shaped by publishers, institutions, and funders intensifies the problem by favoring exciting results over robust ones. Poor quality of biomedical research not only slows down the scientific progress, it also represents an ethical issue. When animal experiments do not contribute to any knowledge gain, the justification for these experiments is not given, and animal lives are wasted. Preregistration of animal research, i.e., registration of the study protocol before the experiment has started, supports scientists in designing an excellent research study, prevents questionable scientific practices, and improves the reporting of all outcomes. With the online platform animalstudyregistry.org, scientists worldwide can now easily register their animal study and show their commitment to open science. To enable a broad uptake of preregistration into biomedical research, funders, publishers, and institutions need to acknowledge preregistration as an engagement for transparent and robust science.

Background Exposure to environmental chemicals that interfere with normal estrogen function can lead to adverse health effects, including cancer. High-throughput screening (HTS) approaches facilitate the efficient identification and characterization of such substances. Objectives We recently described the development of the E-Morph Assay, which measures changes at adherens junctions as a clinically-relevant phenotypic readout for estrogen receptor (ER) alpha signaling activity. Here, we describe its further development and application for automated robotic HTS. Methods Using the advanced E-Morph Screening Assay, we screened a substance library comprising 430 toxicologically-relevant industrial chemicals, biocides, and plant protection products to identify novel substances with estrogenic activities. Based on the primary screening data and the publicly available ToxCast dataset, we performed an in silico similarity search to identify further substances with potential estrogenic activity for follow-up hit expansion screening, and built seven in silico ER models using the conformal prediction (CP) framework to evaluate the HTS results. Results The primary and hit confirmation screens identified 27 ‘known’ estrogenic substances with potencies correlating very well with the published ToxCast ER Agonist Score (r = +0.95). We additionally detected potential ‘novel’ estrogenic activities for 10 primary hit substances and for another nine out of 20 structurally similar substances from in silico predictions and follow-up hit expansion screening. The concordance of the E-Morph Screening Assay with the ToxCast ER reference data and the generated CP ER models was 71% and 73%, respectively, with a high predictivity for ER active substances of up to 87%, which is particularly important for regulatory purposes. Discussion These data provide a proof-of-concept for the combination of in vitro HTS approaches with in silico methods (similarity search, CP models) for efficient analysis of large substance libraries in order to prioritize substances with potential estrogenic activity for subsequent testing against higher tier human endpoints.

Non‐technical summaries of research projects allow tracking the numbers and purpose of animal experiments related to SARS‐CoV2 research so as to provide greater transparency on animal use.

A key challenge of mixture toxicity testing is that a multitude of substances with even more combinations need to be tested in a broad dose range. Consequently testing in rodent bioassays, the current gold standard of toxicity testing, is hardly feasible. High-throughput compatible cell culture systems, however, suffer from limitations with respect to toxicokinetics, tissue interactions, and compensatory mechanisms. Therefore, simple organisms like the nematode Caenorhabditis elegans, combining relevant advantages of complex in vivo and fast in vitro assays might prove highly valuable within a testing strategy for mixtures. To investigate the comparability between results obtained with C. elegans and traditional rodent assays, we used five azole fungicides as well investigated model substances. Our findings suggest that azoles act additively in C. elegans which is in line with previous results in rats. Additionally, we show that toxicokinetics are one important factor for the differences in the relative toxicity of the azoles in both species. Importantly, we also demonstrate that in contrast to most rodent in vivo studies, C. elegans assays provide well-defined concentration-response relationships which are a very good basis for the prediction of mixture effects. We conclude that C. elegans may be an appropriate model for mixture toxicity testing at least within a first step to identify and prioritize relevant mixtures for further testing.

Bei dem Einsatz von pharmakologisch wirksamen Substanzen treffen das Tierversuchsrecht und das Arzneimittelrecht aufeinander. Dieser Beitrag zeigt die in der Tierversuchspraxis in diesem Kontext auftretenden Probleme auf und schlagt praxistaugliche Losungsansatze vor.

25 years after the first Berlin Workshop on Developmental Toxicity this 10th Berlin Workshop aimed to bring together international experts from authorities, academia and industry to consider scientific, methodologic and regulatory aspects in risk assessment of developmental toxicity and to debate alternative strategies in testing developmental effects in the future. Proposals for improvement of the categorization of developmental effects were discussed as well as the update of the DevTox database as valuable tool for harmonization. The development of adverse outcome pathways relevant to developmental neurotoxicity (DNT) was debated as a fundamental improvement to guide the screening and testing for DNT using alternatives to animal methods. A further focus was the implementation of an in vitro mechanism-based battery, which can support various regulatory applications associated with the assessment of chemicals and mixtures. More interdisciplinary and translation research should be initiated to accelerate the development of new technologies to test developmental toxicity. Technologies in the pipeline are (i) high throughput imaging techniques, (ii) models for DNT screening tests, (iii) use of computer tomography for assessment of thoracolumbar supernumerary ribs in animal models, and (iv) 3D biofabrication of bone development and regeneration tissue models. In addition, increased collaboration with the medical community was suggested to improve the relevance of test results to humans and identify more clinically relevant endpoints. Finally, the participants agreed that this conference facilitated better understanding innovative approaches that can be useful for the identification of developmental health risks due to exposure to chemical substances.

Summary Estrogens play an important role in the development and progression of human cancers, particularly in breast cancer. Breast cancer progression depends on the malignant destabilization of adherens junctions (AJs) and disruption of tissue integrity. We found that estrogen receptor alpha (ERα) inhibition led to a striking spatial reorganization of AJs and microclustering of E-Cadherin (E-Cad) in the cell membrane of breast cancer cells. This resulted in increased stability of AJs and cell stiffness and a reduction of cell motility. These effects were actomyosin-dependent and reversible by estrogens. Detailed investigations showed that the ERα target gene and epidermal growth factor receptor (EGFR) ligand Amphiregulin (AREG) essentially regulates AJ reorganization and E-Cad microclustering. Our results not only describe a biological mechanism for the organization of AJs and the modulation of mechanical properties of cells but also provide a new perspective on how estrogens and anti-estrogens might influence the formation of breast tumors., Graphical Abstract, Highlights • ERα inhibition causes adherens junction (AJ) reorganization through AREG and EGFR • AJ reorganization coincides with microclustering of E-Cadherin at cell membranes • AJ reorganization and microclustering of E-Cadherin are actomyosin dependent • AJ reorganization correlates with increased cell stiffness and reduced motility, Cell Biology; Cancer

The Animal Study Registry offers scientists a range of benefits by preregistering their studies. Wider adoption could address the reproducibility problem in biomedical research and enhance animal welfare.

Reliability of data has become a major concern in the course of the reproducibility crisis. Especially when studying animal behavior, confounding factors such as novel test apparatus can lead to a wide variability of data. At worst, effects of novelty or stress related behavior can mask treatment effects and the behavioral data may be misinterpreted. Habituation to the test situation is a common practice to circumvent novelty induced increases in variance and to improve the reliability of the respective measurements. However, there is a lack of published empirical knowledge regarding reasonable habituation procedures and a method validation seems to be overdue.This study aimed at setting up a simple strategy to increase reliability of behavioral data. Therefore, exemplary data from mice tested in an Open Field (OF) arena were used to elucidate the potential of habituation and how reliability of measures can be confirmed by means of a repeatability analysis using the software R. On seven consecutive days, male C57BL/6J, BALB/cJ and 129S1/SvImJ mice were tested in an OF arena once daily and individual mouse behavior (distance travelled, average activity) was recorded. A repeatability analysis was conducted in order to estimate the reliability of measured animal behavior with regard to repeated trials of habituation.Our data analysis revealed that monitoring animal behavior during habituation is important to determine when individual differences of the measurements are stable. Here, the mixed effect model framework proved to be a powerful tool for estimating repeatability values. Repeatability values from distance travelled and average activity increased over the habituation period, revealing that around 60 % of the variance of the data can be explained by individual differences between mice. The first day of habituation was statistically significantly different from the following 6 days in terms of distance travelled and average activity. A habituation period of three days appeared to be sufficient in this study. Overall these results emphasize the importance of habituation and in depth analysis of habituation data to define the correct starting point of the experiment for improving the reliability and reproducibility of experimental data.

Nach der Novellierung des Tierschutzrechts durch den Erlass der Tierschutz-Versuchstierverordnung haben sich einige Unklarheiten bezuglich der Befugnis zur Durchfuhrung der Betaubung von Versuchstieren ergeben. Einige Berufsgruppen, die vor der Novellierung diese Tatigkeiten zumindest unter Aufsicht ausuben durften, konnten nunmehr ausgeschlossen sein. Dies fuhrt nicht nur zu einer grosen Verunsicherung in Kreisen der Betroffenen, sondern konnte auch eine Wettbewerbsverzerrung auf EU-Ebene zu Folge haben. Die Problematik bedarf daher einer eingehenden rechtlichen Prufung und Losungsfindung.

Die Umsetzung der Tierversuchsrichtlinie 2010/63/EU ins deutsche Recht brachte einige Neuerungen mit sich. In den nunmehr drei Jahren seit dem Inkrafttreten des novellierten Tierschutzgesetzes haben sich einige drangende Probleme in der Anwendungspraxis herauskristallisiert, die in diesem Beitrag naher erlautert und rechtlich bewertet werden.

The Animal Study Registry (ASR; www.animalstudyregistry.org) was launched in January 2019 for preregistration of animal studies in order to increase transparency and reproducibility of bioscience research and to promote animal welfare. The registry is free of charge and is designed for exploratory and confirmatory studies within applied science as well as basic and preclinical research. The registration form helps scientists plan their study thoroughly by asking detailed questions concerning study design, methods, and statistics. With registration, the study automatically receives a digital object identifier (DOI) that marks it as intellectual property of the researcher. To accommodate the researchers concerns about theft of ideas, users can restrict the visibility of their registered studies for up to 5 years. The full content of the study becomes publicly accessible at the end of the embargo period. Because the platform is embedded in the infrastructure of the German Federal Government, continuity and data security are provided. By registering a study in the ASR, researchers can show their commitment to transparency and data quality to reviewers and editors, to third-party donors, and to the general public., This Community Page article presents the Animal Study Registry (www.animalstudyregistry.org), which was launched in January 2019 for pre-registration of animal studies in order to increase transparency and reproducibility of bioscience research and to promote animal welfare.

Buprenorphine is a commonly used opioid to treat moderate to severe pain in mice. Although strain differences regarding basal pain sensitivity and the analgesic effect of other opioids have been described for mice, the data for buprenorphine is incomplete. Hence, we investigated basal pain sensitivity and the analgesic effect of buprenorphine (0.42, 4.0 mg·kg−1) in male C57BL/6J, Balb/cJ and 129S1/SvImJ mice using the incremental hot plate. Additionally, we verified single nucleotide polymorphisms in Cytochrome P450 3a (Cyp3a) genes, which encode for enzymes that are relevant for buprenorphine metabolism, and analyzed serum and brain concentrations of buprenorphine and its metabolites. Finally, in a pilot survey we determined μ-opioid receptor (MOR) protein expression in whole brain lysates. Basal pain sensitivity differed significantly between the mouse strains (Balb/cJ > C57BL/6J > 129S1/SvImJ). Additionally, buprenorphine showed a dose- and strain-dependent effect: at a higher dose it led to increased antinociception in C57BL/6J and Balb/cJ mice, whereas in 129S1/SvImJ mice this effect was diminished. Serum and brain concentrations of buprenorphine and its metabolites dose-dependently increased and differed slightly between the strains at the high dose. However, these slight strain differences did not correlate with pain behavior. Furthermore, serum buprenorphine metabolic ratio and distribution of buprenorphine and its metabolites between brain and blood showed no dose- and only some strain-dependent differences independent from nociceptive behavior. Western blot analysis revealed no strain difference in the basal MOR protein expression in brain lysates. Our results indicate that buprenorphine dosing should be determined in a pilot study for the respective mouse strain to optimize pain treatment and to avoid unwanted side effects. The present pharmacokinetic data and the coarse determination of MOR expression do not explain the strain differences in the analgesic effect of buprenorphine. However, follow-up studies focusing on more specific pharmacodynamic factors could further elucidate the reasons.

Background: The growing requirement of hazard and risk assessment of environmental chemicals and the efforts to minimize animal testing, increases the demand for innovative and predictive in vitro test systems in toxicology, reflecting the physiological conditions of human nature. Here, an elemental factor regulating a variety of physiological processes is the day-night rhythm. This circadian rhythm, describing a biological oscillation with a 24-h period is hardly acknowledged in toxicology and test method development. Whilst, in animals or humans the entire organism exhibits a rigorous cellular circadian synchrony, in conventional in vitro systems each cell follows its own rhythm, due to the absence of appropriate synchronizing signals. Objective: Here we investigated whether circadian synchronization of human cells in an in vitro system improves the cellular response and, thus, increases the sensitivity of the test system. Since the circadian regulation of metabolism is particularly well understood, and dioxin and dioxin-like compounds are of major concern for environmental health we focused on the ubiquitous drug metabolizing detoxification system mediated by the aryl hydrocarbon receptor (AHR). Methods: To this end, we applied various prototypical AHR activators onto different human cell lines under non-synchronized or circadian synchronized conditions and determined the dose response on representative endogenous target genes. Results: Remarkably, the cellular response dynamic upon chemical treatment was substantially enhanced in circadian synchronized cells and followed a rhythmic expression pattern. This broader dynamic range was associated with a strikingly higher induction of AHR target genes and the corresponding enzymatic activity, thereby rather mimicking the in vivo situation. Conclusion: Our findings indicate that a synchronized circadian rhythm in a cell culture based test system can improve the physiological relevance of an appropriate in vitro method by reflecting the biological in vivo situation more closely. Accordingly, it is a promising tool to facilitate the wide acceptance of in vitro methods in the field of regulatory toxicology and to further optimize the toxicological assessment of environmental chemicals. Keywords: Circadian rhythm, In vitro, CYP1A1, Environmental chemicals, TCDD, Xenobiotic

Representatives of applied science (e.g. governmental organizations, academia, and industry) met to discuss the progress towards a harmonized human health risk assessment in developmental toxicology of plant protection products, biocidal products, and other environmental chemicals at the 9(th) Berlin Workshop on Developmental Toxicity held in September 2018. Within the focus of the scientific discussion were the future of in-vitro methods for developmental and reproductive toxicology, the potential relevance of alternative species in testing of developmental effects, and risk and hazard assessment of developmental and endocrine effects. Furthermore, the need for a harmonized terminology for classification of anomalies in laboratory animals in developmental toxicity studies aiming for human health risk assessment was determined. Here, the DevTox database was identified as an extremely valuable tool. Overall, the participants agreed that still one of the biggest challenges for testing developmental toxicity in the 21(st) century is the development of animal-free test strategies and alternatives to animal testing that could provide human-relevant information in a rapid, efficient, and mechanistically informative manner.

Summary Growth and offspring count are two commonly determined toxicological endpoints for chemical- or gene-induced developmental and reproductive effects in Caenorhabditis elegans. Here, we present a protocol for a 96 h, medium-throughput assay, assessing both endpoints quantitatively within an automated framework using open-source software. The assay utilizes whole 96-well fluorescence images taken with a high-content screening system. Alternatively, conventional fluorescence images can also be utilized with only a few adjustments. For complete details on the use and execution of this protocol, please refer to Wittkowski et al. (2019)., Graphical Abstract, Highlights • A medium-throughput assay to determine C. elegans growth and offspring count • Applicable to high-content screening and conventional fluorescence microscopy • Automated data acquisition and processing with open-source software, Growth and offspring count are two commonly determined toxicological endpoints for chemical- or gene-induced developmental and reproductive effects in Caenorhabditis elegans. Here, we present a protocol for a 96 h, medium-throughput assay, assessing both endpoints quantitatively within an automated framework using open-source software. The assay utilizes whole 96-well fluorescence images taken with a high-content screening system. Alternatively, conventional fluorescence images can also be utilized with only a few adjustments.

This article is a report of the 8th Berlin Workshop on Developmental Toxicity held in May 2014. The main aim of the workshop was the continuing harmonization of terminology and innovations for methodologies used in the assessment of embryo- and fetotoxic findings. The following main topics were discussed: harmonized categorization of external, skeletal, visceral and materno-fetal findings into malformations, variations and grey zone anomalies, aspects of developmental anomalies in humans and laboratory animals, and innovations for new methodologies in developmental toxicology. The application of Version 2 terminology in the DevTox database was considered as a useful improvement in the categorization of developmental anomalies. Participants concluded that initiation of a project for comparative assessments of developmental anomalies in humans and laboratory animals could support regulatory risk assessment and university-based training. Improvement of new methodological approaches for alternatives to animal testing should be triggered for a better understanding of developmental outcomes.

The cytochrome P450 (CYP) enzyme superfamily is the most important enzyme system for phase I biotransformation. For toxico- and pharmacokinetic studies, use of liver-based microsomes, including those of mice, is state-of-the-art to study CYP-dependent metabolism. However, reproducibility and interpretation of these data is still very variable, partly because current testing guidelines do not cover details on organ sampling and potential liver lobe differences. Hence, we analyzed CYP activity, CYP protein content, mRNA expression of CYP1A, CYP2C, CYP2D and CYP3A isozymes, and cytochrome P450 reductase (CPR) activity of the four different liver lobes and processus papillaris of male C57BL/6J mice in comparison to whole liver. Additionally, we used whole liver of Balb/cJ and 129S1/SvImJ for strain comparison. Our data show significant differences in CYP activity, being most prominent in lobus sinister lateralis and lobus medialis, and lowest in processus papillaris. These differences were not caused by varying Cyp gene expression or CYP protein level, but partly correspond with lobe specific CPR activities. We also observed significant strain differences in CYP mRNA expression and activities with overall high activities in 129S1/SvImJ mice and low activities in Balb/cJ mice compared to C57BL/6J mice. In addition, strain specific differences in CYP2C and CYP2D activity seem to be reflected in strain dependent differences in CPR activity. In summary, our results indicate that in mice CYP activity and gene expression are strain dependent and may vary highly between liver lobes. To ensure reproducibility and comparability of different probes and studies, this should be taken into account when liver samples are collected for the analysis of CYP-dependent metabolism.

Automatic extraction of semantic relations from text can support finding relevant information from scientific publications. We describe our participation in Task 7 of SemEval-2018 for which we experimented with two relations extraction tools - jSRE and TEES - for the extraction and classification of six relation types. The results we obtained with TEES were significantly superior than those with jSRE (33.4% vs. 30.09% and 20.3% vs. 16%). Additionally, we utilized the model trained with TEES for extracting semantic relations from biomedical abstracts, for which we present a preliminary evaluation.

We developed a new probabilistic model to assess the impact of recommendations rectifying the reproducibility crisis (by publishing both positive and 'negative' results and increasing statistical power) on competing objectives, such as discovering causal relationships, avoiding publishing false positive results, and reducing resource consumption. In contrast to recent publications our model quantifies the impact of each single suggestion not only for an individual study but especially their relation and consequences for the overall scientific process. We can prove that higher-powered experiments can save resources in the overall research process without generating excess false positives. The better the quality of the pre-study information and its exploitation, the more likely this beneficial effect is to occur. Additionally, we quantify the adverse effects of both neglecting good practices in the design and conduct of hypotheses-based research, and the omission of the publication of 'negative' findings. Our contribution is a plea for adherence to or reinforcement of the good scientific practice and publication of 'negative' findings.

The xenoestrogens Bisphenol-A (BPA) and Nonylphenol (NP) are endocrine disruptors used in the plastic polymer industry to manufacture different products for human use. Previous studies have suggested a role of these compounds in the shedding of signaling molecules, such as tumor necrosis factor α (TNF-α). The aim of this work was to evaluate the effect of BPA and NP on the sheddase ADAM17 and its newly discovered regulators iRhom1 and iRhom2 in the release of EGFR-ligands. We report that BPA and NP can stimulate the release of the ADAM17-substrates HB-EGF and TGF-α. In cells lacking ADAM17 (Adam17(−/−) mEFs) BPA-stimulated release of HB-EGF, but not TGF-α, was strongly reduced, whereas NP-stimulated shedding of HB-EGF and TGF-α was completely abolished. Inactivation of both ADAM17 and the related ADAM10 (Adam10/17(−/−) mEFs) completely prevented the release of these substrates. In the absence of iRhom1, BPA- or NP-stimulated release of HB-EGF or TGF-α was comparable to wild type control mEFs, conversely the BPA-induced release of HB-EGF was abolished in iRhom2(−/−) mEFs. The defect in shedding of HB-EGF in iRhom2(−/−) mEF cells could be rescued by overexpressing iRhom2. Interestingly, the NP-stimulated release of HB-EGF was not affected by the absence of iRhom2, suggesting that NP could potentially activate both ADAM10 and ADAM17. We tested this hypothesis using betacellulin (BTC), an EGFR-ligand that is a substrate for ADAM10. We found that NP, but not BPA stimulated the release of BTC in Adam17(−/−), iRhom2(−/−) or iRhom1/2(−/−), but not in Adam10/17(−/−) cells. Taken together, our results suggest that BPA and NP stimulate the release of EGFR-ligands by differentially activating ADAM17 or ADAM10. The identification of specific effects of these endocrine disruptors on ADAM10 and ADAM17 will help to provide a better understanding of their roles in cell signaling and proinflammatory processes, and provide new potential targets for treatment of reproductive or inflammatory diseases such as asthma or breast cancer that are promoted by xenoestrogens.

In the European Union (EU), animal welfare is seen as a matter of great importance. However, with respect to animal experimentation, European citizens feel quite uninformed. The European Directive 2010/63/EU for the protection of laboratory animals aims for greater transparency and requires that a comprehensible, nontechnical summary (NTS) of each authorised research project involving animals is published by the respective Member State. However, the NTSs remain sleeping beauties if their contents are not easily and systematically accessible. The German web-based NTS database AnimalTestInfo is a unique channel for scientists to communicate their work, and provides the opportunity for large-scale analyses of planned animal studies to inform researchers and the public. For an in-depth meta-analysis, we classified the duly completed NTSs submitted to AnimalTestInfo in 2014 and 2015 according to the International Classification of Diseases and Related Health Problems (ICD) system. Indexing the NTSs with ICD codes provided a fine-grained overview of the prospective uses of experimental animals. Using this approach, transparency, especially for highly controversial animal research involving, for example, nonhuman primates, is fostered, as it enables pinpointing the envisaged beneficiary down to the level of the addressed disease. Moreover, research areas with many planned projects involving animals can be specified in detail. The development of 3R (replacement, reduction, and refinement) measures in these research areas may be most efficient, as a large number of experimental animals would benefit from it. Indexing NTSs with ICD codes can support governments and funding agencies in advancing target-oriented funding of 3R research. Data drawn from NTSs can provide a basis for the development, validation, and implementation of directed 3R strategies as well as guidance for rethinking the role of animal research models., Author summary The AnimalTestInfo database was developed in Germany to make the nontechnical summaries (NTSs) of animal research studies available in a searchable and easily accessible web-based format. This database helps address requirements stipulated in a European Directive regarding the transparency and publication of animal experiments. In this meta-analysis, we analysed NTSs submitted in 2014 and 2015 to AnimalTestInfo and classified them according to the International Classification of Diseases and Related Health Problems (ICD) system. Each NTS was assigned an ICD-10 code based on the envisioned beneficiary of the research project. Based on these codes, we conducted various quantitative analyses. Our results provide a granular overview of the prospective uses of experimental animals and identify areas in need of alternative strategies to help replace, reduce, and refine animal research. We hope this data will inform governments and funding agencies in advancing the integrity and reporting of responsible animal research.

Assessing the impact of recommendations rectifying the reproducibility crisis (publishing both positive and “negative” results and increasing statistical power) on competing objectives, such as discovering causal relationships, avoiding publishing false positive results, and reducing resource consumption, we developed a new probabilistic model. Our model quantifies the impact of each single suggestion for an individual study and their relation and consequences for the entire research process. We can prove that higher-powered experiments can save resources in the overall research process without generating excess false positives. The better the quality of the pre-study information and its exploitation, the more likely this beneficial effect is to occur. Additionally, we quantify the adverse effects of neglecting good practices in the design and conduct of hypotheses-based research, and the responsibility to publish “negative” findings. Our contribution is a plea for adherence to or reinforcement of the good scientific practice and publication of “negative” findings.

Many European citizens see animal welfare a matter of great importance [1]. Initiatives such as Stop Vivisection , which petitions politicians to abandon all support for animal experimentation in biomedical and toxicological research (http://ec.europa.eu/citizens-initiative/public/initiatives/successful/details/2012/000007/en?lg=en), might suggest that a majority of Europeans want to see the use of all animals in testing and research banned. However, a recent Eurobarometer survey on Science and Technology implies a different attitude: Although public opinion is divided when it comes to the use of dogs and monkeys in animal testing, a vast majority accept the use of mice in research if it produces new information on human health problems [2]. > … further scientific knowledge is needed to reliably and efficiently evaluate phenotypes and to sufficiently apply refinement, which will guarantee a high standard of animal welfare These different perspectives on animal experimentation are reflected in the European Directive 2010/63/EU on the protection of animals used for scientific purposes [3]. On the one hand, the Directive clearly spells out a final goal of fully replacing all procedures that use live animals. On the other hand, it also recognizes that animal testing is still necessary for basic research and to protect human and animal health and the environment (Recital 10). Another key element of the Directive is the 3R principle—replace, reduce, refine—which seeks not only to ultimately replace all animal testing, but in the meantime to reduce the number of animals being used and to refine their use in experiments. > Together, all parties have to find a solution that assures a high level of legal security, which can be implemented easily by both scientists and competent authorities It was a well‐considered decision by the European Commission to enact a directive rather …