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Animals used in biomedical research may experience pain or distress as part of the experimental protocol. By implementing humane endpoints, pain or distress can be prevented or alleviated whilst still meeting scientific aims and objectives. We invited experts from Arizona State University (Samantha Sullivan) and Loma Linda University (Obed Rutebuka), along with USDA and OLAW representatives (Louis DiVincenti and Axel Wolff), to respond to a scenario about the importance of defining humane endpoints in research protocols.