Addressing Distress and Pain in Animal Research: The Veterinary, Research, Societal, Regulatory and Ethical Contexts for Moving Forward

While most people recognize that biomedical scientists are searching for knowledge that will improve the health of humans and animals, the image of someone deliberately causing harm to an animal in order to produce data that may lead to some future benefit has always prompted an uncomfortable reaction outside the laboratory. However, proponents of animal research have usually justified the practice by reference to greater benefits (new knowledge and medical treatments) over lesser costs (in animal suffering and death). Given that one of the costs of animal research is the suffering experienced by the animals, the goal of eliminating distress and pain in the animal laboratory wherever feasible, is one that few, if any, people (especially scientists) would argue against. Moreover, we contend that a laudable, long-range goal would be the elimination of all substantial distress and pain in the animal laboratory (Stephens and Conlee, 2004) although some might see this as an insurmountable technical challenge

The Minimization of Research Animal Distress and Pain: Conclusions and Recommendations

While the attention given to preventing, assessing, and alleviating pain in research animals has increased noticeably in recent decades, much remains to be done both in terms of implementing best practices and conducting studies to answer outstanding questions. In contrast, the attention to distress (particularly non-pain induced distress) has shown no comparable increase. There are many reasons for this discrepancy, including the conceptual untidiness of the distress concept, the paucity of pharmacological treatments for distress, and perceived lack of regulatory emphasis on distress. These are challenges that need to be addressed and overcome. This book is intended to help meet these and other challenges to effectively tackling distress and pain in research animals. The chapter, in particular, distills the various recommendations regarding recognition, assessment, measurement and alleviation of animal distress and pain throughout this book, in order to provide the reader with practical information in a succinct format

Addressing Distress and Pain in Animal Research: The Veterinary, Research, Societal, Regulatory and Ethical Contexts for Moving Forward

While most people recognize that biomedical scientists are searching for knowledge that will improve the health of humans and animals, the image of someone deliberately causing harm to an animal in order to produce data that may lead to some future benefit has always prompted an uncomfortable reaction outside the laboratory. However, proponents of animal research have usually justified the practice by reference to greater benefits (new knowledge and medical treatments) over lesser costs (in animal suffering and death). Given that one of the costs of animal research is the suffering experienced by the animals, the goal of eliminating distress and pain in the animal laboratory wherever feasible, is one that few, if any, people (especially scientists) would argue against. Moreover, we contend that a laudable, long-range goal would be the elimination of all substantial distress and pain in the animal laboratory (Stephens and Conlee, 2004) although some might see this as an insurmountable technical challenge

The First Forty Years of the Alternatives Approach: Refining, Reducing, and Replacing the Use of Laboratory Animals

The concept of the Three Rs— reduction, refinement, and replacement of animal use in biomedical experimentation—stems from a project launched in 1954 by a British organization, the Universities Federation for Animal Welfare (UFAW). UFAW commissioned William Russell and Rex Burch to analyze the status of humane experimental techniques involving animals. In 1959 these scientists published a book that set out the principles of the Three Rs, which came to be known as alternative methods. Initially, Russell and Burch’s book was largely ignored, but their ideas were gradually picked up by the animal protection community in the 1960s and early ’70s. In the ’80s, spurred by public pressure, the alternatives approach was incorporated into national legislation throughout the developed countries and embraced by industry in Europe and America. Government centers devoted to the validation and regulatory acceptance of alternative methods were established during the ’90s. By 2000 the use of animals in research had fallen by up to fifty percent from its high in the 1970s

Possibilities for Refinement and Reduction: Future Improvements Within Regulatory Testing

Approaches and challenges to refining and reducing animal use in regulatory testing are reviewed. Regulatory testing accounts for the majority of animals reported in the most painful and/or distressful categories in the United States and Canada. Refinements in testing, including the use of humane endpoints, are of increasing concern. Traditional approaches to reduction (e.g., improving experimental design) are being supplemented with complementary approaches, such as the use of tier testing to eliminate some chemicals prior to in vivo testing. Technological advances in telemetry and noninvasive techniques will help decrease either the demand for animals in testing or animal suffering. Further decreases in animal use will stem from international harmonization and coordination of testing programs. Progress in refinement and reduction faces a variety of broad challenges, including limited funding for research. In the specific area of refinement, a key challenge is the issue of distress (as distinct from pain). In the area of reduction, the practice of using unjustifiably high numbers of animals from small species (e.g., rodents) should be challenged. One case study of the use of carbon dioxide as a euthanasia agent illustrates the need for further analysis and research. Notwithstanding the complexities and challenges, the potential for refinement and reduction in regulatory testing is encouraging

Carbon Dioxide for Euthanasia: Concerns Regarding Pain and Distress, with Special Reference to Mice and Rats

Carbon dioxide (CO2) is the most commonly used agent for euthanasia of laboratory rodents, used on an estimated tens of millions of laboratory rodents per year worldwide, yet there is a growing body of evidence indicating that exposure to CO2 causes more than momentary pain and distress in these and other animals. We reviewed the available literature on the use of CO2 for euthanasia (as well as anaesthesia) and also informally canvassed laboratory animal personnel for their opinions regarding this topic. Our review addresses key issues such as CO2 flow rate and final concentration, presence of oxygen, and prefilled chambers (the animal is added to the chamber once a predetermined concentration and flow rate have been reached) versus gradual induction (the animal is put into an empty chamber and the gas agent(s) is gradually introduced at a fixed rate). Internationally, animal research standards specify that any procedure that would cause pain or distress in humans should be assumed to do so in nonhuman animals as well (Public Health Service 1986, US Department of Agriculture 1997, Organization for Economic Cooperation and Development 2000). European Union guidelines, however, specify a certain threshold of pain or distress, such as ‘skilled insertion of a hypodermic needle’, as the starting point at which regulation of the use of animals in experimental or other scientific procedures begins (Biotechnology Regulatory Atlas n.d.). There is clear evidence in the human literature that CO2 exposure is painful and distressful, while the non-human literature is equivocal. However, the fact that a number of studies do conclude that CO2 causes pain and distress in animals indicates a need for careful reconsideration of its use. Finally, this review offers recommendations for alternatives to the use of CO2 as a euthanasia agent

Combination of hydrogel nanoparticles and proteomics to reveal secreted proteins associated with decidualization of human uterine stromal cells

<p>Abstract</p> <p>Background</p> <p>Identification of secreted proteins of low abundance is often limited by abundant and high molecular weight (MW) proteins. We have optimised a procedure to overcome this limitation.</p> <p>Results</p> <p>Low MW proteins in the conditioned media of cultured cells were first captured using dual-size exclusion/affinity hydrogel nanoparticles and their identities were then revealed by proteomics.</p> <p>Conclusions</p> <p>This technique enables the analysis of secreted proteins of cultured cells low MW and low abundance.</p

Targeting Leader Cells in Ovarian Cancer as an Effective Therapeutic Option

Majority of ovarian cancers are diagnosed at advanced stages with intra-peritoneal spread as the most common mode of disease metastasis. The formation of cancer spheroids is essential for the collective migration process, where shed tumour cells from the primary tumour form aggregates rather than disseminating as individual cells and seed within the peritoneal cavity. These cancer spheroids consist of leader cells (LC) and follower cells (FC), with the LC subset as key drivers of cellular movement and invasion. LCs have stem cell-like properties and are highly chemo-resistant with a specific survival addiction to several cell signalling pathways, such as the PI3K/AKT/mTOR pathway. We explore in this book chapter, the evidence supporting the role of LC in OC metastasis and the suppression of LC as an attractive therapeutic option for the treatment of advanced OC

Resolving Animal Distress and Pain: Principles and Examples of Good Practice in Various Fields of Research

Pain and distress are central topics in legislation, regulations, and standards regarding the use of animals in research. However, in practice, pain has received greatly increased attention in recent years, while attention to distress has lagged far behind, especially for distress that is not induced by pain. A contributing factor is that there is less information readily available on distress, including practical information on its recognition, assessment and alleviation. This chapter attempts to help fill that void by reversing the usual pattern and giving greater attention to distress than to pain. In addition, we also bypass the pain versus distress dichotomy by adopting a holistic treatment of adverse effects, i.e., not parsing distress and pain, by providing guidance on how to assess deviations from normality through tools such as score sheets. Our aim is to provide practical information to IACUCs, scientists, technicians and animal care personnel. We organize the chapter according to specific research areas and case studies. However, the principles and approaches are readily generalized to other research areas. CONTENTS Effect of surgical technical skill on pain and distress in animals - Alicia Karas, DVM Carbon dioxide euthanasia: example of aversion techniques - Matthew C. Leach, PhD The Refinement of Infectious Disease Research - Karl A. Andrutis DVM, MS, DACLAM Polyclonal antibody production - Kathleen Conlee, BS, MPA Animal models of human psychopathology: anxiety - John P. Gluck PhD Refinement In Toxicology Testing: A Workshop to Promote Current Advances and Disseminate Best Practices - Andrew N. Rowan, Martin L. Stephens, and Kathleen M. Conle

Current Status of Endovascular Training for Cardiothoracic Surgery Residents in the United States

Background Endovascular interventions for cardiovascular pathology are becoming increasingly relevant to cardiothoracic surgery. This study assessed the perceived prevalence and efficacy of endovascular skills training and identified differences among training paradigms. Methods Trainee responses to questions in the 2016 In-Service Training Examination survey regarding endovascular training were analyzed based on the four different cardiothoracic surgery training pathways: traditional 2- and 3-year thoracic, integrated 6-year, and combined 4+3 general and thoracic residency programs. Results The duration of endovascular training was substantially different among programs, at a median of 17 weeks for integrated 6-year, 8.5 weeks for 3-year, 6 weeks for 4+3, and 4 weeks for 2-year residency (p < 0.0001). After adjusting for year of training and program type, the duration of endovascular rotations was significantly associated with self-assessed comfort with catheter-based skills (p < 0.0001). Eighty-two percent of residents rotated with trainees from other specialties, and 58% experienced competition for cases. Residents reported greater exposure to transcatheter aortic valve replacement than to thoracic endovascular aortic repair, cardiac catheterization, percutaneous closure of atrial septal defect, and transcatheter mitral valve surgery (p < 0.0001). A significant proportion of responders reported feeling uncomfortable performing key steps of transcatheter aortic valve replacement (52%) or thoracic endovascular aortic repair (49%). Conclusions Considerable heterogeneity exists in endovascular training among cardiothoracic surgery training pathways, with a significant number of residents having minimal to no exposure to these emerging techniques. These findings highlight the need for a standardized curriculum to improve endovascular exposure and training