Poor Translatability of Biomedical Research Using Animals - A Narrative Review.

The failure rate for the translation of drugs from animal testing to human treatments remains at over 92%, where it has been for the past few decades. The majority of these failures are due to unexpected toxicity - that is, safety issues revealed in human trials that were not apparent in animal tests - or lack of efficacy. However, the use of more innovative tools, such as organs-on-chips, in the preclinical pipeline for drug testing, has revealed that these tools are more able to predict unexpected safety events prior to clinical trials and so can be used for this, as well as for efficacy testing. Here, we review several disease areas, and consider how the use of animal models has failed to offer effective new treatments. We also make some suggestions as to how the more human-relevant new approach methodologies might be applied to address this.

Severity classification of surgical procedures and application of health monitoring strategies in animal research proposals: A retrospective review.

Animal experimentation has been one of the most controversial areas of animal use, mainly due to the intentional harms inflicted upon the animals used. In an effort to reduce these harms, research on refinement has increased significantly over the past 20 years. However, the extent to which these efforts have helped to reduce the severity of the research procedures, and thus animal suffering, is uncertain. To provide an indication of the awareness and implementation of refinement methods, we reviewed the experimental techniques for 684 surgical interventions described in 506 animal research applications that had been sent to the German competent authorities for approval in 2010. In this paper, we describe and discuss the severity categorisation of the proposed surgeries and the planned health monitoring strategies. We found that the researchers frequently underestimated the levels of pain, suffering, distress and lasting harm that were to be inflicted on the animals. Furthermore, the planned health monitoring strategies were generally flawed. To ensure responsible treatment of animals and high-quality science, adequate training of research workers in recognising and alleviating animal suffering is essential.

The application of humane endpoints and humane killing methods in animal research proposals: A retrospective review.

Refinement refers to the use of methods that help to minimise animal suffering in the laboratory. Research in this area has increased significantly over the past two decades. However, the extent to which refinements are applied in practice is uncertain. To provide an indication of the implementation and awareness of refinements, we reviewed the experimental techniques for 684 surgical interventions described in 506 animal research applications sent to the German competent authorities for approval in 2010. In this paper, we describe and discuss the appropriateness of the proposed humane endpoints and killing methods. We found that, when the investigators included humane endpoints in their application, these were often lacking in detail and/or were to be implemented at a late stage of suffering. In addition, the choice of method to kill the animals could be improved in the majority of the applications. We provide recommendations for future improvements, based on the recent literature. To ensure scientific rigour, avoid needless animal suffering and enable an accurate harm-benefit analysis, animal researchers have to be knowledgeable about refinement methods and apply them effectively. To assess compliance and ensure that only those studies in which potential benefits outweigh the harms are carried out, reviews such as ours - as well as retrospective assessments of actual harms and benefits - should be conducted widely and regularly, and the findings should be published.

Limitations of Particle-Based Spasers.

We present a semiclassical analytic model for spherical core-shell surface plasmon lasers. Within this model, we drop the widely used one-mode approximations in favor of fully electromagnetic Mie theory. This allows for incorporation of realistic gain relaxation rates that so far are massively underestimated. Especially, higher order modes can undermine and even reverse the beneficial effects of the strong Purcell effect in such systems. Our model gives a clear view on gain and resonator requirements, as well as on the output characteristics that will help experimenters to design more efficient particle-based spasers.

Complementary and alternative medicine (CAM) as part of primary health care in Germany-comparison of patients consulting general practitioners and CAM practitioners: a cross-sectional study.

BACKGROUND: In Germany, complementary and alternative medicine (CAM) in primary health care is offered by general practitioners (GPs) and by natural health practitioners, so called 'Heilpraktiker' (HPs). Considering the steadily growing number of unregulated HPs, the aim of the study was to assess characteristics of patients consulting HPs in comparison to patients consulting GPs. METHODS: In a cross-sectional study, patients of randomly selected GPs and HPs were asked to complete a questionnaire about their health care status, health care behavior, and symptoms rated on the Measure Yourself Medical Outcome Profile (MYMOP-D). Patient groups were compared based on health care provider (HP, GP with high use of CAM (CAM-GP), and GP with no/little use of CAM (nCAM-GP)) using Kruskal-Wallis tests and analyses of variance (ANOVA). RESULTS: Altogether, 567 patients (91 of 11 HPs, 223 of 15 CAM-GPs, 253 of 19 nCAM-GPs) filled in the questionnaire. Patients of HPs had a higher education level and were more often female. The most common reason for encounter among all three groups were musculoskeletal problems (30.2-31.1 %). Patients seeing HPs reported more psychological (4.4 % vs. 17.8 %), but less respiratory problems (19.9 % vs. 7.8 %), and longer symptom duration (>5 years: 21.1 % vs. 40.7 %), than patients of nCAM-GPs. CONCLUSIONS: The high percentage of patients with psychological illness and chronic health problems consulting HPs demonstrates the urgent need for action with regard to CAM therapy in primary care and regulation of natural health practitioners. Appropriate measures with regard to quality and patient safety should be taken given the growing numbers of HPs and the absence of a regulatory body.

A brief patient-reported outcome instrument for primary care: German translation and validation of the Measure Yourself Medical Outcome Profile (MYMOP).

BACKGROUND: Measure Yourself Medical Outcome Profile (MYMOP) is a patient-generated outcome instrument capable of measuring effects from a wide range of health care interventions. This paper reports the translation of this instrument into German (MYMOP-D) and the assessment of validity and sensitivity to change for the MYMOP-D. The instrument was piloted in a German primary care context. METHODS: The translation process was conducted according to international guidelines. Recruited patients of both general practitioners and non-medical Complementary and Alternative Medicine (CAM) practitioners ("Heilpraktiker") in the German state of Baden-Wuerttemberg completed a questionnaire comprised of the MYMOP-D and the EQ-5D. Responses were analysed to assess construct validity. For assessing the instrument's sensitivity to change, patients received the MYMOP-D again after four weeks at which point they were also asked for their subjective views on change of symptoms. Correlation between MYMOP-D and EQ-5D and sensitivity to change as gradient in score change and as standardized response mean (SRM) were calculated. RESULTS: 476 patients from general practices and 91 patients of CAM practitioners were included. Construct validity of the MYMOP-D was given with a correlation of r = .47 with the EQ-5D. Sensitivity to change for subjective change of symptoms could only be analysed for improvement or no change of symptoms, as only 12 patients reported deterioration of symptoms. Results showed the expected smooth gradient with 2.2, 1.3, and 0.5 points of change for large, little improvement and no change, respectively. SRM for MYMOP-D Profile Score was 0.88. CONCLUSIONS: The MYMOP-D shows excellent construct validity. It is able to detect changes when symptoms in patients improve or remain unchanged. Deterioration of symptoms could not be evaluated due to too few data. With its brevity and simplicity, it might be an important tool for enhancing patient-centred care in the German health care context.

Effects of pramlintide in patients with type 2 diabetes mellitus: an analysis using daily insulin dose tertiles.

OBJECTIVE: The purpose of the analysis was to investigate if the efficacy and tolerability of 6 months of pramlintide therapy in patients with type 2 diabetes mellitus (T2DM) differed with increasing levels of concomitant insulin doses, using data from 3 previously described clinical trials. METHODS: In this post hoc analysis, data from 2 pooled, placebo-controlled pivotal trials and 1 clinical practice trial were evaluated by baseline insulin use tertile in patients with T2DM. RESULTS: In the pivotal trials, both glycated hemoglobin (A1C) and body weight decreased similarly across tertiles with pramlintide. A1C decreased slightly and body weight remained relatively unchanged across tertiles with placebo. Similarly, in the clinical practice trial, pramlintide was associated with decreases in A1C, body weight, and total daily insulin use across the tertiles. Overall, the most common adverse events were gastrointestinal in nature, and the rate of severe hypoglycemia was low. CONCLUSION: These results suggest that pramlintide therapy was associated with improved A1C and decreased body weight, with a low rate of severe hypoglycemia, among patients with T2DM, regardless of baseline insulin use.

60 Years of the 3Rs symposium: Lessons learned and the road ahead.

When The Principles of Humane Experimental Technique was published in 1959, authors William Russell and Rex Burch had a modest goal: to make researchers think about what they were doing in the laboratory - and to do it more humanely. Sixty years later, their groundbreaking book was celebrated for inspiring a revolution in science and launching a new field: The 3Rs of alternatives to animal experimentation. On November 22, 2019, some pioneering and leading scientists and researchers in the field gathered at the Johns Hopkins Bloomberg School of Public Health in Bal-timore for the 60 Years of the 3Rs Symposium: Lessons Learned and the Road Ahead. The event was sponsored by the Johns Hopkins Center for Alternatives to Animal Testing (CAAT), the Foundation for Chemistry Research and Initiatives, the Alternative Research & Development Foundation (ARDF), the American Cleaning Institute (ACI), the International Fragrance Association (IFRA), the Institute for In Vitro Sciences (IIVS), John "Jack" R. Fowle III, and the Society of Toxicology (SoT). Fourteen pres-entations shared the history behind the groundbreaking publication, international efforts to achieve its aims, stumbling blocks to progress, as well as remarkable achievements. The day was a tribute to Russell and Burch, and a testament to what is possible when people from many walks of life - science, government, and industry - work toward a common goal.

William Russell and Rex Burch published their book The Principles of Humane Experimental Technique in 1959. The book encouraged researchers to replace animal experiments where it was possible, to refine experiments with animals in order to reduce their suffering, and to reduce the number of animals that had to be used for experiments to the minimum. Sixty years later, a group of pioneering and leading scientists and researchers in the field gathered to share how the publi­cation came about and how the vision inspired international collaborations and successes on many different levels including new laws. The paper includes an overview of important milestones in the history of alternatives to animal experimentation.

First Organoid Intelligence (OI) workshop to form an OI community.

The brain is arguably the most powerful computation system known. It is extremely efficient in processing large amounts of information and can discern signals from noise, adapt, and filter faulty information all while running on only 20 watts of power. The human brain's processing efficiency, progressive learning, and plasticity are unmatched by any computer system. Recent advances in stem cell technology have elevated the field of cell culture to higher levels of complexity, such as the development of three-dimensional (3D) brain organoids that recapitulate human brain functionality better than traditional monolayer cell systems. Organoid Intelligence (OI) aims to harness the innate biological capabilities of brain organoids for biocomputing and synthetic intelligence by interfacing them with computer technology. With the latest strides in stem cell technology, bioengineering, and machine learning, we can explore the ability of brain organoids to compute, and store given information (input), execute a task (output), and study how this affects the structural and functional connections in the organoids themselves. Furthermore, understanding how learning generates and changes patterns of connectivity in organoids can shed light on the early stages of cognition in the human brain. Investigating and understanding these concepts is an enormous, multidisciplinary endeavor that necessitates the engagement of both the scientific community and the public. Thus, on Feb 22-24 of 2022, the Johns Hopkins University held the first Organoid Intelligence Workshop to form an OI Community and to lay out the groundwork for the establishment of OI as a new scientific discipline. The potential of OI to revolutionize computing, neurological research, and drug development was discussed, along with a vision and roadmap for its development over the coming decade.

Engagement of scientists with the public and policymakers to promote alternative methods.

Scientists are usually good at teaching, sometimes even to lay audiences. But communicating with journalists, activists, or policymakers can be a different story - hesitancy to make mistakes as well as the temptation to disproportionally promote one's own case come into play. The multitude of social media and other web-based outlets has diversified and accelerated the communication of science. Real-time reactions, sharing of data, tools and results, increasing invitation of personal opinion, demand for transparency, political correctness, and loss of trust in experts are challenges to researchers in general. The field of alternatives to animal testing is more political and important to lay audiences than many others, so its scientists must be especially aware of these challenges. Public engagement offers the opportunity to form community and create wide support for non-animal research and its implementation. This requires scientists to step out of the ivory tower of higher education and engage with diverse interest groups by outreach activities, interviews, and press releases, etc. by employing tailored communication.

The future of Parkinson's disease research: A new paradigm of human-specific investigation is necessary and possible.

Parkinson's disease (PD) is a complex neurodegenerative condition with a multifactorial origin. To date, approaches to drug discovery for PD have resulted in symptomatic therapies for the motor manifestations and signs associated with neurodegeneration but have failed to identify preventive or curative therapies. This failure mainly originates from the persistence of major gaps in our understanding of the specific molecular basis of PD initiation and progression. New approach methodologies (NAMs) hold the potential to advance PD research while facilitating a move away from ani-mal-based research. We report a workshop involving NAM experts in the field of PD and neurodegenerative diseases, who discussed and identified a scientific strategy for successful, human-specific PD research. We outline some of the most important human-specific NAMs, along with their main potentials and limitations, and suggest possible ways to overcome the latter. Key recommendations to advance PD research include integrating NAMs while accounting for multiple levels of complexity, from molecular to population level; learning from recent advances in Alzheimer's disease research; increasing the sharing of data; promoting innovative pilot studies on disease pathogenesis; and accessing philanthropic funding to enable studies using novel approaches. Collaborative efforts between different stakeholders, including researchers, clinicians, and funding agencies, are urgently needed to create a scientific roadmap and support a paradigm change towards effective, human-specific research for neurodegenerative diseases without animals, as is already happening in the field of toxicology.

New European Union statistics on laboratory animal use - what really counts!

Seven years after the last release, the European Commission has again collated and released data on laboratory animal use. The new report is the first to correspond to the requirements of the new Directive 2010/63/EU. Beside minor problems in reporting, the new reporting format is a major step forward, with additional new categories like severity allowing insight into animal use related questions that goes far beyond the previous reports. An in-depth analysis confirms a slight decrease in animal use from 2015 to 2017, but also compared to the 2005, 2008 and 2011 reports, though the new reporting scheme makes this comparison difficult. Notable success is evident for replacing rabbit pyrogen testing but, in general, the implementation of accepted alternative methods lags behind expec-tations. Beside the roughly 10 million animals per year covered in the report, about 8 million animals were identified that fall under the Directive but are not included in this number. Their omission downplays the impact of REACH on animal use. The report, second to none in its detail internationally, represents an important instrument for benchmarking and strategi-cally focusing activities in the 3Rs.

Diabetes mellitus and gastric emptying: questions and issues in clinical practice.

It is long known that both type 1 and type 2 diabetes can be associated with changes in gastric emptying; a number of publications have linked diabetes to delayed gastric emptying of variable severity and often with poor relationship to gastrointestinal symptomatology. In contrast, more recent studies have reported accelerated gastric emptying when adjusted for glucose concentration in patients with diabetes, indicating a reciprocal relationship between gastric emptying and ambient glucose concentrations. This review proposes that gastroparesis or gastroparesis diabeticorum, a severe condition characterized by a significant impairment of gastric emptying accompanied by severe nausea, vomiting, and malnutrition, is often overdiagnosed and not well contrasted with delays in gastric emptying. The article offers a clinically relevant definition of gastroparesis that should help differentiate this rare condition from (often asymptomatic) delays in gastric emptying. The fact that delayed gastric emptying can also be observed in non-diabetic individuals under experimental conditions in which hyperglycaemia is artificially induced suggests that a delay in gastric emptying rate when blood glucose concentrations are high is actually an appropriate physiological response to hyperglycaemia, slowing further increases in blood glucose. The article discusses the strengths and weaknesses of various methodologies for assessing gastric emptying, especially with respect to the diabetes population, and reviews newer diabetes therapies that decelerate the rate of gastric emptying. These therapies may be a beneficial tool in managing postprandial hyperglycaemia because they attenuate rapid surges in glucose concentrations by slowing the delivery of meal-derived glucose.

Retrospective review of anesthetic and analgesic regimens used in animal research proposals.

Pain has a profound effect on an animal's wellbeing. In Germany, researchers using animals have been legally required since 1972 to reduce any possible pain, suffering, distress or lasting harm to an absolute minimum. To evaluate how these provisions have been implemented in practice, an assessment of refinements to experimental techniques was conducted by retrospectively reviewing 684 surgical interventions described in 506 animal research applications that were sent to the German competent authorities for approval in 2010. This paper focuses on the efficacy of proposed anesthesia and peri- and postoperative analgesia. Postoperative analgesia was not proposed for 30 % of surgeries. Following 10 % of procedures, animals were to be given pain relieving medication if the investigators decided this was necessary; however, structured assessments to detect pain were absent. Consequences of unalleviated pain and omission of pain assessment techniques are discussed, and some recommendations to improve anesthesia and analgesia are given. The findings of this review highlight the need for improvement, both to fulfil legal requirements and to improve animal welfare. To monitor compliance with animal welfare regulations, and ensure good veterinary and scientific practices, education and training needs to be intensified. Adherence to the items listed in the PREPARE and ARRIVE guidelines and the Gold Standard Publication checklist (GSPC) should become legally binding.

Beyond the 3Rs: Expanding the use of human-relevant replacement methods in biomedical research.

This year marks the 60th anniversary of Russell and Burch's pioneering book, The Principles of Humane Experimental Technique. Their 3Rs framework has helped to inspire humane and scientific progress in experimental technique. However, it is time to update its strategic application. The 21st century has already seen the development of promising, high-tech non-animal models, such as organs-on-a-chip and computational approaches that, in our view, will replace animals as the default option in biomedical experimentation. How fast this transition will take place will depend on the pace at which these new models are optimized to reflect the biology of humans, rather than that of non-human animals. While the new methods are likely to reshape all areas in which animals are currently used in science, we particularly encourage their application in biomedical research, which accounts for the bulk of animals used. We call for the pursuit of a three-prong strategy that focuses on (1) advancing non-animal methods as replacements of animal experiments, (2) applying them to biomedical research, and (3) improving their relevance to human biology. As academics and scientists, we feel that educational efforts targeted at young scientists in training will be an effective and sustainable way to advance this vision. Our strategy may not promise an imminent end to the use of animals in science, but it will bring us closer to an era in which the 3Rs are increasingly perceived as a solution to a receding problem. Russell and Burch themselves surely would have welcomed these positive changes.

Animal Research, Accountability, Openness and Public Engagement: Report from an International Expert Forum.

In November 2013, a group of international experts in animal research policy (n = 11) gathered in Vancouver, Canada, to discuss openness and accountability in animal research. The primary objective was to bring together participants from various jurisdictions (United States, Sweden, Australia, New Zealand, Germany, Canada and the United Kingdom) to share practices regarding the governance of animals used in research, testing and education, with emphasis on the governance process followed, the methods of community engagement, and the balance of openness versus confidentiality. During the forum, participants came to a broad consensus on the need for: (a) evidence-based metrics to allow a "virtuous feedback" system for evaluation and quality assurance of animal research, (b) the need for increased public access to information, together with opportunities for stakeholder dialogue about animal research, (c) a greater diversity of views to be represented on decision-making committees to allow for greater balance and (d) a standardized and robust ethical decision-making process that incorporates some sort of societal input. These recommendations encourage aspirations beyond merely imparting information and towards a genuine dialogue that represents a shared agenda surrounding laboratory animal use.

Effect of adjunctive pramlintide treatment on treatment satisfaction in patients with type 1 diabetes.

OBJECTIVE: To assess the effect of adjunctive pramlintide treatment on treatment satisfaction in patients with type 1 diabetes treated with intensive insulin regimens. RESEARCH DESIGN AND METHODS: Intensively treated (multiple daily injection [MDI] or continuous subcutaneous insulin infusion [CSII] pump therapy) patients with type 1 diabetes completed a study-specific treatment satisfaction questionnaire following 29 weeks of either placebo (n = 136) or pramlintide (n = 130) treatment in a double-blind, noninferiority pramlintide dose titration trial. End points included patient reported outcomes, their relationship to insulin treatment regimen, A1C, weight, and insulin use. RESULTS: Pramlintide-treated patients reported greater treatment satisfaction in most questionnaire responses. Treatment satisfaction was similar for pramlintide-treated patients regardless of intensive insulin regimens (MDI versus CSII). Mean A1C was reduced to a similar degree in both pramlintide- (-0.39 +/- 0.07%) and placebo-treated (-0.45 +/- 0.07%) patients. However, pramlintide treatment was associated with reductions in mean body weight (-1.50 +/- 0.33 kg; P < 0.0001) and mealtime insulin use (-19.05 +/- 5.17%; P < 0.005) over 29 weeks, while placebo treatment resulted in weight gain (1.28 +/- 0.25 kg) and a smaller reduction in mealtime insulin use (-2.20 +/- 3.33%). CONCLUSIONS: Despite similar reductions in A1C, pramlintide treatment resulted in greater treatment satisfaction compared with placebo treatment. This was independent of insulin delivery method.

Towards a 21st-century roadmap for biomedical research and drug discovery: consensus report and recommendations.

Decades of costly failures in translating drug candidates from preclinical disease models to human therapeutic use warrant reconsideration of the priority placed on animal models in biomedical research. Following an international workshop attended by experts from academia, government institutions, research funding bodies, and the corporate and non-governmental organisation (NGO) sectors, in this consensus report, we analyse, as case studies, five disease areas with major unmet needs for new treatments. In view of the scientifically driven transition towards a human pathways-based paradigm in toxicology, a similar paradigm shift appears to be justified in biomedical research. There is a pressing need for an approach that strategically implements advanced, human biology-based models and tools to understand disease pathways at multiple biological scales. We present recommendations to help achieve this.

Role of endogenous amylin in glucagon secretion and gastric emptying in rats demonstrated with the selective antagonist, AC187.

Amylin is a 37-amino acid polypeptide co-secreted with insulin from the pancreatic beta-cells. It complements insulin's stimulation of the rate of glucose disappearance (Rd) by slowing the rate of glucose appearance (Ra) through several mechanisms, including an inhibition of mealtime glucagon secretion and a slowing of gastric emptying. To determine if endogenous amylin tonically inhibits these processes, we studied the effects of the amylin receptor blocker AC187 upon glucagon secretion during euglycemic, hyperinsulinemic clamps in Sprague-Dawley (HSD) rats, upon gastric emptying in HSD rats, and upon gastric emptying and plasma glucose profile in hyperamylinemic, and genetically obese, Lister Albany/NIH rats during a glucose challenge. Amylin blockade increased glucagon concentration, accelerated gastric emptying of liquids, and resulted in an exaggerated post-challenge glycemia. These data collectively indicate a physiologic role for amylin in glucose homeostasis via mechanisms that include regulation of glucagon secretion and gastric emptying.