bluuebunny/crossref_metadata_2025 huggingface dataset:
Toward Preclinical Predictive Drug Testing for Metabolism and Hepatotoxicity by Using In Vitro Models Derived from Human Embryonic Stem Cells and Human Cell Lines — A Report on the Vitrocellomics EU-project
Authors: Mandenius Carl-Fredrik , Andersson Tommy B. , Alves Paula M. , Batzl-Hartmann Christine , Björquist Petter , Carrondo Manuel J.T. , Chesne Christophe , Coecke Sandra , Edsbagge Josefina , Fredriksson J. Magnus , Gerlach Jörg C. , Heinzle Elmar , Ingelman-Sundberg Magnus , Johansson Inger , Küppers-Munther Barbara , Müller-Vieira Ursula , Noor Fozia , Zeilinger Katrin
Published: April 2019

Drug-induced liver injury is a common reason for drug attrition in late clinical phases, and even for post-launch withdrawals. As a consequence, there is a broad consensus in the pharmaceutical industry, and within regulatory authorities, that a significant improvement of the current in vitro test methodologies for accurate assessment and prediction of such adverse effects is needed. For this purpose, appropriate in vivo-like hepatic in vitro models are necessary, in addition to novel sources of human hepatocytes. In this report, we describe recent and ongoing research toward the use of human embryonic stem cell (hESC)-derived hepatic cells, in conjunction with new and improved test methods, for evaluating drug metabolism and hepatotoxicity. Recent progress on the directed differentiation of human embryonic stem cells to the functional hepatic phenotype is reported, as well as the development and adaptation of bioreactors and toxicity assay technologies for the testing of hepatic cells. The aim of achieving a testing platform for metabolism and hepatotoxicity assessment, based on hESC-derived hepatic cells, has advanced markedly in the last 2–3 years. However, great challenges still remain, before such new test systems could be routinely used by the industry. In particular, we give an overview of results from the Vitrocellomics project (EU Framework 6) and discuss these in relation to the current state-of-the-art and the remaining difficulties, with suggestions on how to proceed before such in vitro systems can be implemented in industrial discovery and development settings and in regulatory acceptance.
Assessing the Search for Information on Three Rs Methods, and their Subsequent Implementation: A National Survey among Scientists in the Netherlands
Authors: van Luijk Judith , Cuijpers Yvonne , van der Vaart Lilian , Leenaars Marlies , Ritskes-Hoitinga Merel
Published: April 2019

A local survey conducted among scientists into the current practice of searching for information on Three Rs (i.e. Replacement, Reduction and Refinement) methods has highlighted the gap between the statutory requirement to apply Three Rs methods and the lack of criteria to search for them. To verify these findings on a national level, we conducted a survey among scientists throughout The Netherlands. Due to the low response rate, the results give an impression of opinions, rather than being representative of The Netherlands as a whole. The findings of both surveys complement each other, and indicate that there is room for improvement. Scientists perceive searching the literature for information on Three Rs methods to be a difficult task, and specific Three Rs search skills and knowledge of Three Rs databases are limited. Rather than using a literature search, many researchers obtain information on these methods through personal communication, which means that published information on possible Three Rs methods often remains unfound and unused. A solution might be to move beyond the direct search for information on Three Rs methods and choose another approach. One approach that seems rather appropriate is that of systematic review. This provides insight into the necessity for any new animal studies, as well as optimal implementation of available data and the prevention of unnecessary animal use in the future.
An Alternative Gas-phase In Vitro Exposure System for Toxicity Testing: The Interaction Between Nitrous Oxide and A549 Cells
Authors: Stellavato Antonietta , Cammarota Marcella , Miraglia Nadia , Simonelli Angela , Giuliano Mariateresa
Published: April 2019

An original in vitro approach was adopted to expose cells to volatile agents. The anaesthetic nitrous oxide (N2O) was chosen as the model agent, and type II pneumocyte-like cells (A549 cells) were used as the target to represent the lungs. A time-lapse microscopy station was equipped with a manual gas mixer that allowed the generation of a mixture of N2O/air/CO2 in the gas phase, to provide a uniform distribution of the volatile agent. The dissolution of N2O in the culture medium was monitored by gas chro-matography–electron capture detection. Biochemical alterations, in terms of homocysteine accumulation, demonstrated that intracellular methionine synthase had been inactivated by N2O absorbed by the cells, a process that also occurs in vivo. Toll-like receptors, which are key molecules in inflammatory lung diseases, were also investigated at the molecular level. Our experiments indicated that biochemical and molecular alterations occurred in the cells, even under conditions where neither morphologic changes nor consistent alterations in cell proliferation were evident. This in vitro exposure system can be efficiently adopted for looking at the repeat-dose effects of volatile agents on respiratory tissues. Moreover, it could be of further benefit for identifying the wide range of specific cell targets, and for monitoring relevant endpoints in the cellular and molecular processes that occur during exposure to volatile compounds.
A Critical Evaluation of the 2011 ECHA Reports on Compliance with the REACH and CLP Regulations and on the Use of Alternatives to Testing on Animals for Compliance with the REACH Regulation
Authors: Spielmann Horst , Sauer Ursula G. , Mekenyan Ovanes
Published: April 2019

On 30 June 2011, the European Chemicals Agency published two reports, one on the functioning of the REACH system, the other on the use of alternatives to animal testing in compliance with that system. The data presented are based on information gained during the first registration period under the REACH system, which included high production volume chemicals and substances of very high concern, which have the most extensive information requirements. A total of 25,460 registration dossiers were received, covering 3,400 existing, so-called ‘phase-in’, substances, and 900 new, so-called ‘non-phase-in’, substances. Data sharing and the joint submission of data are reported to have worked successfully. In the registration dossiers for these substances, results from new animal tests were included for less than 1% of all the endpoints; testing proposals (required for ‘higher-tier’ information requirements) were submitted for 711 in vivo tests involving vertebrate animals. The registrants mainly used old, existing experimental data, or options for the adaptation (waiving) of information requirements, before collecting new information. For predicting substance toxicity, ‘read-across’ was the second most-used approach, followed by ‘weight-of-evidence’. In vitro toxicity tests played a minor role, and were only used when the respective test methods had gained the status of regulatory acceptance. All in all, a successful start to the REACH programme was reported, particularly since, in contrast to most predictions, it did not contribute to a significant increase in toxicity testing in animals.
Testing the Biocompatibility of a Glutathione-containing Intraocular Irrigation Solution by Using an Isolated Perfused Bovine Retina Organ Culture Model — an Alternative to Animal Testing
Authors: Januschowski Kai , Zhour Ahmad , Lee Albert , Maddani Ramin , Mueller Sebastian , Spitzer Martin S. , Schnichels Sven , Schultheiss Maximilian , Doycheva Deshka , Bartz-Schmidt Karl-Ulrich , Szurman Peter
Published: April 2019

The effects of a glutathione-containing intraocular irrigation solution, BSS Plus®, on retinal function and on the survival of ganglion cells in whole-mount retinal explants were studied. Evidence is provided that the perfused ex vivo bovine retina can serve as an alternative to in vivo animal testing. Isolated bovine retinas were prepared and perfused with an oxygen-saturated standard irrigation solution, and an electroretinogram was recorded to assess retinal function. After stable b-waves were detected, the isolated retinas were perfused with BSS Plus for 45 minutes. To investigate the effects of BSS Plus on photoreceptor function, 1mM aspartate was added to the irrigation solution in order to obtain a-waves, and the ERG trace was monitored for 75 minutes. For histological analysis, isolated whole retinal mounts were stored for 24 hours at 4°C, in the dark. The percentages of cell death in the retinal ganglion cell layer and in the outer and inner nuclear layers were estimated by using an ethidium homodimer-1 stain and the TUNEL assay. General swelling of the retina was examined with high-resolution optical coherence tomography. During perfusion with BSS Plus, no significant changes in a-wave and b-wave amplitudes were recorded. Retinas stored for 24 hours in BSS Plus showed a statistically significant smaller percentage (52.6%, standard deviation [SD] = 16.1%) of cell death in the retinal ganglion cell layer compared to the control group (69.6%, SD = 3.9, p = 0.0031). BSS Plus did not seem to affect short-term retinal function, and had a beneficial effect on the survival of retinal ganglion cells. This method for analysing the isolated perfused retina represents a valuable alternative for testing substances for their retinal biocompatibility and toxicity.
Validation Study of the In Vitro Skin Irritation Test with the LabCyte EPI-MODEL24
Authors: Kojima Hajime , Ando Yoko , Idehara Kenji , Katoh Masakazu , Kosaka Tadashi , Miyaoka Etsuyoshi , Shinoda Shinsuke , Suzuki Tamie , Yamaguchi Yoshihiro , Yoshimura Isao , Yuasa Atsuko , Watanabe Yukihiko , Omori Takashi
Published: April 2019

A validation study on an in vitro skin irritation assay was performed with the reconstructed human epidermis (RhE) LabCyte EPI-MODEL24, developed by Japan Tissue Engineering Co. Ltd (Gamagori, Japan). The protocol that was followed in the current study was an optimised version of the EpiSkin protocol (LabCyte assay). According to the United Nations Globally Harmonised System (UN GHS) of classification for assessing the skin irritation potential of a chemical, 12 irritants and 13 non-irritants were validated by a minimum of six laboratories from the Japanese Society for Alternatives to Animal Experiments (JSAAE) skin irritation assay validation study management team (VMT). The 25 chemicals were listed in the European Centre for the Validation of Alternative Methods (ECVAM) performance standards. The reconstructed tissues were exposed to the chemicals for 15 minutes and incubated for 42 hours in fresh culture medium. Subsequently, the level of interleukin-1 alpha (IL-1α) present in the conditioned medium was measured, and tissue viability was assessed by using the MTT assay. The results of the MTT assay obtained with the LabCyte EPI-MODEL24 (LabCyte MTT assay) demonstrated high within-laboratory and between-laboratory reproducibility, as well as high accuracy for use as a stand-alone assay to distinguish skin irritants from non-irritants. In addition, the IL-1α release measurements in the LabCyte assay were clearly unnecessary for the success of this model in the classification of chemicals for skin irritation potential.
A Review of Autopsy Reports on Chimpanzees in or from US Laboratories
Authors: Capaldo Theodora , Peppercorn Marge
Published: April 2019

Approximately 1000 chimpanzees are currently held in five federally owned, or supported, US laboratories. This study reviews 110 autopsy reports on chimpanzees who died from 2001–2011 in laboratories or in sanctuaries (but who were from laboratories), in order to glean information about their pre-morbid health and causes of death. The findings raise questions about the health status of the chimpanzees remaining in laboratories. Most of the chimpanzees currently held are not involved in active protocols. The Chimpanzee Health Improvement, Maintenance, and Protection (CHIMP) Act 2000 states that chimpanzees “not needed” for research “shall” be accepted into the federal sanctuary system, but criteria for when a chimpanzee is deemed “not needed” are not given. The assessment of “not needed” lies with the Secretary of Health and Human Services, who has left the decision to the discretion of the laboratories. This autopsy review revealed that the majority of the chimpanzees who died in laboratories had been suffering from significant chronic or incurable illnesses, and most often had multi-system diseases that should have made them ineligible for future research, on scientific, as well as ethical, grounds. The study's findings are significant in establishing the need for defined criteria for chimpanzee retirement to sanctuary.
Animal use in Veterinary Education — The Need for a Fourth R: Respect
Authors: Tiplady Catherine
Published: April 2019

Animals should not be treated with disrespect, merely because they are surplus to the requirements of others
Toward a Humanised Alternative to the Use of Laboratory Animals for Blood–Brain Barrier Research
Authors: Patabendige Adjanie
Published: April 2019

The development of a novel flow-based, three-dimensional human blood–brain barrier model for studying brain infections is now under way
Adverse Outcome Pathway-based Screening Strategies for an Animal-free Safety Assessment of Chemicals
Authors: Landesmann Brigitte , Mennecozzi Milena , Berggren Elisabet , Whelan Maurice
Published: April 2019

Currently, the assessment of risk to human health from exposure to manufactured chemicals is mainly based on experiments performed on living animals ( in vivo). Substantial efforts are being undertaken to develop alternative solutions to in vivo toxicity testing. This new paradigm, based on the Mode-of-Action (MoA) framework, postulates that any adverse human health effect caused by exposure to an exogenous substance can be described by a series of causally-linked biochemical or biological key events with measurable parameters. The elaboration of mechanistic knowledge through literature research is necessary for a MoA-driven design of integrated testing strategies using in vitro methods for in vivo predictions. The objective of our ongoing research is to demonstrate the feasibility of an integrated approach to predict human toxicity following the Adverse Outcome Pathway (AOP) framework. In our previous work on MoA with the HepaRG cell model, we developed a strategy to identify chemicals that were hepatotoxic. This pioneered an innovative way of using data from in vitro experiments to group chemicals based on their MoA, which is likely to be an important step in a toxicity testing strategy.
A Proposed Study on the Transplacental Transport of Parabens in the Human Placental Perfusion Model
Authors: Mathiesen Line , Zuri Giuseppina , Andersen Maria H. , Knudsen Lisbeth E.
Published: April 2019

Human exposure to parabens as a preservative used in personal care products is of increasing concern, as there is evidence from in vivo and in vitro studies of hormone disruption in association with exposure to parabens. Transport across the placenta could be critical for risk assessment, but the available data are sparse. The aim is to develop a method for estimating fetal exposure, via the placenta, to the most commonly-used parabens, by using a human placental perfusion model. The use of human tissue is vital for determining human fetal exposure, because animal studies are of little relevance, since the placenta exhibits significant interspecies variation. An HPLC model is currently being established to simultaneously quantify four different parabens, namely, methylparaben, ethylparaben, propylparaben and butylparaben, and their main metabolite, p-hydroxybenzoic acid. With this model, we aim to determine the transport kinetics of these parabens across the human placenta, and to investigate placental metabolism, including differences in transport due to molecular characteristics. This will facilitate assessment of the risks associated with the use of paraben-containing products during pregnancy.
Cryopreserved Human Whole Blood: A Human-based In Vitro Immunotoxicological System
Authors: Rivera-Mariani Félix E.
Published: April 2019

The airborne environment poses numerous human health risks, including potential exposure to particulate matter and to fungal bioaerosols with pro-inflammatory capabilities. The utility of animal models in the study of the human health effects of these exposures is often limited. This is due to the necessity for a high-throughput of samples and the high financial costs involved, as well as various associated animal welfare issues. In addition, animal models might not appropriately predict the responses of humans to pro-inflammatory stimuli. Here are discussed the recent applications of a human-based assay, the cryopreserved human whole blood system, to study the human health effects of exposures to particulate matter and to the less-studied fungal spores of higher fungi (phylum Basidiomycota or ‘true mushrooms’). Evidence is provided to support the utility of the human-based assay in the fields of environmental health and medical mycology, for the evaluation of common paradigms in environmental health studies, and to highlight the potential human health effects of spores of basidiomycete fungi.
8. UFAW and Major Charles Hume
Authors: Balls Michael
Published: April 2019

UFAW and its founder, Major Charles W. Hume, played a vital role in providing an opportunity for William Russell and Rex Burch to develop the concepts which are now widely appreciated as The Principles of Humane Experimental Technique
Chlorotriazines Do Not Activate the Aryl Hydrocarbon Receptor, the Oestrogen Receptor or the Thyroid Receptor in In Vitro Assays
Authors: de la Casa-Resino Irene , Navas José M. , Fernández-Cruz María L.
Published: April 2019

Atrazine, prometryn, propazine and simazine are chlorotriazines that are commonly employed as herbicides. However, their use is a major cause of concern, due to their reported endocrine disrupting effects in different taxa. Data from studies on the molecular and cellular processes underlying the hormonal action of these substances are contradictory. The ability of these chlorotriazines and the atrazine metabolites, desethyl-s-chlorotriazine and desisopropyl-s-chlorotriazine, to trigger responses mediated by the oestrogen receptor (ER), aryl hydrocarbon receptor (AhR) and thyroid receptor (TR), was studied by using in vitro approaches. Transcriptional activation assays were applied to observe the activation of ER and TR. The induction of ethoxyresorufin- O-deethylase (EROD) activity in the RTG-2 cell line served as an indicator of AhR activation. No responses were found in any of the assays, with any of the six chlorotriazines tested. Our observations indicate that the chlorotriazines tested are unlikely to cause their endocrine effects via these receptors.
Mechanistically-based QSARs to Describe Metabolic Constants in Mammals
Authors: Pirovano Alessandra , Huijbregts Mark A.J. , Ragas Ad M.J. , Veltman Karin , Hendriks A. Jan
Published: April 2019

Biotransformation is one of the processes which influence the bioaccumulation of chemicals. The enzymatic action of metabolism involves two processes, i.e. the binding of the substrate to the enzyme followed by a catalytic reaction, which are described by the Michaelis–Menten constant (Km) and the maximum rate (Vmax). Here, we developed Quantitative Structure–Activity Relationships (QSARs) for Log(1/Km) and LogVmax for substrates of four enzyme classes. We focused on oxidations catalysed by alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), flavin-containing monooxygenase (FMO) and cytochrome P450 (CYP) in mammals. The chemicals investigated were xenobiotics, including alcohols, aldehydes, pesticides and drugs. We applied general linear models for this purpose, employing descriptors related to partitioning, geometric characteristics, and electronic properties of the substrates, which can be interpreted mechanistically. The explained variance of the QSARs varied between 20% and 70%, and it was larger for Log(1/Km) than for LogVmax. The increase of 1/Km with compound logP and size suggests that weak interactions are important, e.g. by substrate binding via desolvation processes. The importance of electronic factors for 1/Km was described in relation to the catalytic mechanism of the enzymes. Vmax was particularly influenced by electronic properties, such as dipole moment and energy of the lowest unoccupied molecular orbital. This can be explained by the nature of the catalysis, characterised by the cleavage and formation of covalent or ionic bonds (strong interactions). The present study may be helpful to understand the underlying principles of the chemical specific activity of four important oxidising enzymes.
Ethoxyresorufin-O-deethylase (EROD) Activity Modulation of 2,3,7,8-Tetrachlorodibenzo-p-dioxin and 3,3′,4,4′,5-Pentachlorobiphenyl (PCB 126) in the Presence of Aqueous Suspensions of Nano-C60
Authors: Rathore Rajesh , Schramm Karl-Werner
Published: April 2019

The increase in commercial production and inevitable release of fullerenes into the environment accelerates concerns about their potential toxicity. Furthermore, the concomitant release of xenobiotics poses a health hazard to humans, and might present potential long-term risks to human health. In the present study, we found that an aqueous suspension of buckminsterfullerene (aqu-nC60) does not result in the induction of ethoxyresorufin- O-deethylase (EROD) activity in H4IIE rat liver cells in vitro. The simultaneous and sequential exposure of aqu-nC60 and the dioxin TCDD induces EROD activity to the same extent as TCDD alone (i.e. in the absence of fullerene), in spite of the high affinity of C60 for TCDD. However, the co-exposure of aqu-nC60 and PCB 126 induces elevated EROD activity, and sequential exposure increases responses 2-fold compared to the control samples. Our in vitro observations suggest a potential source of drug–drug type interaction of fullerene with xenobiotics, particularly after a sequential exposure.
The Virtual Pharmacology Lab — A Repository of Free Educational Resources to Support Animal-free Pharmacology Teaching
Authors: Dewhurst David , Ward Ross
Published: April 2019

The new repository will support pharmacology practical teaching by promoting the sharing and re-use of existing pharmacology resources
Student Perspectives on the Use of Alternative Methods for Teaching in Veterinary Faculties
Authors: Sachana Magda , Theodoridis Alexandros , Cortinovis Cristina , Pizzo Fabiola , Kehagias Evaggelos , Albonico Marco , Caloni Francesca
Published: April 2019

The use of alternative methods for teaching purposes is gradually increasing in higher education. In order to evaluate the usefulness of non-animal based practical classes in veterinary science, and to inform on possible benefits and limitations of these teaching tools, a questionnaire was designed and distributed to students. Although there was no complete agreement among the student responses, it was apparent that the majority of the students would like traditional training methods to be paired with alternative approaches, and expressed their desire to be exposed to as many humane modes of learning as possible. In addition, the students agreed that alternative teaching methods for training in veterinary science can reinforce existing knowledge that is required at the clinical stage, and that they can be effective supplements to traditional training methods. It was also concluded from the study that the use of new alternative approaches is very much appreciated by the students, whereas the validity and effectiveness of these methods are debatable, suggesting that further optimisation, proper application and evaluation of these alternative methods is required.
Statistics of Scientific Procedures on Living Animals 2013: Experimentation Continues to Rise — The Reliance on Genetically-altered Animals Must be Addressed
Authors: Hudson-Shore Michelle
Published: April 2019

The 2013 Statistics of Scientific Procedures on Living Animals reveal that the level of animal experimentation in Great Britain continues to rise, with 4.12 million procedures being conducted. The figures indicate that this is almost exclusively a result of the breeding and use of genetically-altered (GA) animals (i.e. genetically-modified animals, plus those with harmful genetic defects). The breeding of GA animals increased to over half (51%) of all the procedures, and GA animals were involved in 61% of all the procedures. Indeed, if these animals were removed from the statistics, the number of procedures would actually have declined by 4%. It is argued that the Coalition Government has failed to address this issue, and, as a consequence, will not be able to deliver its pledge to reduce animal use in science. Recent publications supporting the need to reassess the dominance of genetic alteration are also discussed, as well as the need to move away from the use of dogs as the default second species in safety testing. The general trends in the species used, and the numbers and types of procedures, are also reviewed. Finally, forthcoming changes to the statistics are discussed.
A ‘Road Map’ toward Ending Severe Suffering of Animals Used in Research and Testing
Authors: Lilley Elliot , Hawkins Penny , Jennings Maggy
Published: April 2019

Ending severe suffering is a desirable goal for both ethical and scientific reasons. The RSPCA has pledged to work toward the end of such suffering for laboratory animals, and in this article we outline a practical approach that establishments can follow to achieve this aim
A Normal and Biotransforming Model of the Human Bronchial Epithelium for the Toxicity Testing of Aerosols and Solubilised Substances
Authors: Prytherch Zoë C. , BéruBé Kelly A.
Published: April 2019

In this article, we provide an overview of the experimental workflow by the Lung and Particle Research Group at Cardiff University, that led to the development of the two in vitro lung models — the normal human bronchial epithelium (NHBE) model and the lung–liver model, Metabo-Lung™. This work was jointly awarded the 2013 Lush Science Prize. The NHBE model is a three-dimensional, in vitro, human tissue-based model of the normal human bronchial epithelium, and Metabo-Lung involves the co-culture of the NHBE model with primary human hepatocytes, thus permitting the biotransformation of inhaled toxicants in an in vivo-like manner. Both models can be used as alternative test systems that could replace the use of animals in research and development for safety and toxicity testing in a variety of industries (e.g. the pharmaceutical, environmental, cosmetics, and food industries). Metabo-Lung itself is a unique tool for the in vitro detection of toxins produced by reactive metabolites. This 21st century animal replacement model could yield representative in vitro predictions for in vivo toxicity. This advancement in in vitro toxicology relies on filter-well technology that will enable a wide-spectrum of researchers to create viable and economic alternatives for respiratory safety assessment and disease-focused research.
Animal Use in Education in India: Confusion Continues
Authors: Badyal Dinesh K.
Published: April 2019

In India, there is an urgent need to get a single directive on the appropriate and relevant use of animals in education and research which is equally applicable in all institutions
14. On the Proper Application of Appropriate Statistical Methods
Authors: Balls Michael
Published: April 2019

The proper application of appropriate statistical methods is crucial to relevant and reliable biomedical research and testing
The European Citizens’ Stop Vivisection Initiative
Authors: Balls Michael
Published: April 2019

The European Commission's considerate response to the controversial European Citizens’ Stop Vivisection Initiative promises to accelerate progress toward eliminating the need for animal experimentation in research and testing
The Ex Vivo Eye Irritation Test as an Alternative Test Method for Serious Eye Damage/Eye Irritation
Authors: Spöler Felix , Kray Oya , Kray Stefan , Panfil Claudia , Schrage Norbert F.
Published: April 2019

Ocular irritation testing is a common requirement for the classification, labelling and packaging of chemicals (substances and mixtures). The in vivo Draize rabbit eye test (OECD Test Guideline 405) is considered to be the regulatory reference method for the classification of chemicals according to their potential to induce eye injury. In the Draize test, chemicals are applied to rabbit eyes in vivo, and changes are monitored over time. If no damage is observed, the chemical is not categorised. Otherwise, the classification depends on the severity and reversibility of the damage. Alternative test methods have to be designed to match the classifications from the in vivo reference method. However, observation of damage reversibility is usually not possible in vitro. Within the present study, a new organotypic method based on rabbit corneas obtained from food production is demonstrated to close this gap. The Ex Vivo Eye Irritation Test (EVEIT) retains the full biochemical activity of the corneal epithelium, epithelial stem cells and endothelium. This permits the in-depth analysis of ocular chemical trauma beyond that achievable by using established in vitro methods. In particular, the EVEIT is the first test to permit the direct monitoring of recovery of all corneal layers after damage. To develop a prediction model for the EVEIT that is comparable to the GHS system, 37 reference chemicals were analysed. The experimental data were used to derive a three-level potency ranking of eye irritation and corrosion that best fits the GHS categorisation. In vivo data available in the literature were used for comparison. When compared with GHS classification predictions, the overall accuracy of the three-level potency ranking was 78%. The classification of chemicals as irritating versus non-irritating resulted in 96% sensitivity, 91% specificity and 95% accuracy.
Non-animal Replacements for Acute Toxicity Testing
Authors: Barker-Treasure Carol , Coll Kevin , Belot Nathalie , Longmore Chris , Bygrave Karl , Avey Suzanne , Clothier Richard
Published: April 2019

Current approaches to predicting adverse effects in humans from acute toxic exposure to cosmetic ingredients still heavily necessitate the use of animals under EU legislation, particularly in the context of the REACH system, when cosmetic ingredients are also destined for use in other industries. These include the LD50 test, the Up-and-Down Procedure and the Fixed Dose Procedure, which are regarded as having notable scientific deficiencies and low transferability to humans. By expanding on previous in vitro tests, such as the animal cell-based 3T3 Neutral Red Uptake (NRU) assay, this project aims to develop a truly animal-free predictive test for the acute toxicity of cosmetic ingredients in humans, by using human-derived cells and a prediction model that does not rely on animal data. The project, funded by Innovate UK, will incorporate the NRU assay with human dermal fibroblasts in animal product-free culture, to generate an in vitro protocol that can be validated as an accepted replacement for the currently available in vivo tests. To date, the project has successfully completed an assessment of the robustness and reproducibility of the method, by using sodium lauryl sulphate (SLS) as a positive control, and displaying analogous results to those of the original studies with mouse 3T3 cells. Currently, the testing of five known ingredients from key groups (a surfactant, a preservative, a fragrance, a colour and an emulsifier) is under way. The testing consists of initial range-finding runs followed by three valid runs of a main experiment with the appropriate concentration ranges, to generate IC50 values. Expanded blind trials of 20 ingredients will follow. Early results indicate that this human cell-based test holds the potential to replace aspects of in vivo animal acute toxicity testing, particularly with reference to cosmetic ingredients.
Single Housing of Primates in US Laboratories: A Growing Problem with Shrinking Transparency
Authors: Chandna Alka , Niebo Michael , Lopresti-Goodman Stacy , Goodman Justin
Published: April 2019

Thirty years ago, the United States took steps to enhance the psychological well-being of primates in laboratories, including the introduction of social housing requirements. Now, in an apparent response to questions about the effectiveness of these measures, federal authorities are completely shutting down public access to information on the implementation of social housing
In Vitro Models to Mimic the Endothelial Barrier
Authors: Barbe Laurent , Alini Mauro , Herrmann Marietta , Verrier Sophie
Published: April 2019

Microfluidic technologies permit the replication in vitro of geometrical features essential for the homeostasis of all vascularised tissues in vivo, including the contribution of pericytes to the endothelial barrier
The 2014 Lush Prize Awards
Authors: BéruBé Kelly A. , Redmond Craig
Published: April 2019

The third year of the Lush Prize was perhaps the most successful yet
The ‘Genomic Revolution’ and its Impact on Medical Research
Authors: Chandaria Rehma
Published: April 2019

The sequencing of the human genome held great promise for better disease treatment and a concomitant reduction in animal experiments. Neither promise has been adequately fulfilled
Stakeholder Views on the Creation and Use of Genetically-Engineered Animals in Research
Authors: Ormandy Elisabeth H.
Published: April 2019

This interview-based study examined the diversity of views relating to the creation and use of genetically-engineered (GE) animals in biomedical science. Twenty Canadian participants (eight researchers, five research technicians and seven members of the public) took part in the interviews, in which four main themes were discussed: a) how participants felt about the genetic engineering of animals as a practice; b) governance of the creation and use of GE animals in research, and whether current guidelines are sufficient; c) the Three Rs ( Replacement, Reduction, Refinement) and how they are applied during the creation and use of GE animals in research; and d) whether public opinion should play a greater role in the creation and use of GE animals. Most of the participants felt that the creation and use of GE animals for biomedical research purposes (as opposed to food purposes) is acceptable, provided that tangible human health benefits are gained. However, obstacles to Three Rs implementation were identified, and the participants agreed that more effort should be placed on engaging the public on the use of GE animals in research.
The Perception of Animal Experimentation Ethics among Indian Teenage School Pupils
Authors: Kim Justin Namuk , Choi Eun Hee , Kim Soo-Ki
Published: April 2019

To promote awareness of animal experimentation ethics among teenagers, we created an educational pamphlet and an accompanying questionnaire. One hundred Indian teenage school pupils were given the pamphlet and subsequently surveyed with the questionnaire, to evaluate: a) their perception of animal experimentation ethics; and b) their opinion on the effectiveness of the pamphlet, according to gender and school grade/age. There was a significant correlation between grade/age and support for animal experimentation, i.e. senior students were more inclined to show support for animal experimentation. There was also a significant correlation between gender and perception of the need to learn about animal experimentation ethics, with girls more likely to feel the need to learn about ethics than boys. In addition, the four questions relating to the usefulness of the pamphlet, and student satisfaction with its content, received positive responses from the majority of the students. Even though the pamphlet was concise, it was apparent that three quarters of the students were satisfied with its content. Gender and age did not influence this level of satisfaction. Overall, our study shows that there is a significant correlation between a pupil's school grade/age and their support for animal experimentation, and that there is also a significant correlation between gender and the perceived need to learn about animal experimentation ethics. This pilot scheme involving an educational pamphlet and questionnaire could be beneficial in helping to formulate basic strategies for educating teenage school pupils about animal ethics.
Launch of the Alliance for Human Relevant Science
Authors: Ram Rebecca
Published: April 2019

The Alliance is an exciting new collaboration, founded to address an urgent need to drive research and development, policymaking, awareness, outreach, and education into human-based methods of safety testing and biomedical research
Brazil Starts to Ban Animal Use in Higher Education: A Positive and Progressive Development
Authors: de Cássia Maria Garcia Rita , Jukes Nick , Bones Vanessa , Gebara Rosangela , de Almeida Souza Mariângela Freitas , Ruiz Valeska Regina Reque , Alonso Luciano , Tréz Thales , Oliveira Simone Tostes , Rocha Alexandro Aluisio , Alves Gutemberg , Paixão Rita Leal , Menezes Rita de Cassia Alves Alcântara , Dias Claudia , Andersen Monica Levy , Gasparetto Débora , Capilé Karynn , Matera Júlia Maria , Bachinski Róber
Published: April 2019

The Brazilian government has published a resolution that bans animal use in some practical classes within undergraduate and high school technical education from April 2019. Resolution No. 38/2018, issued by the National Council for the Control of Animal Experimentation (CONCEA), bans the killing of animals for dissection purposes and animal experiments in practical classes that do not involve the acquisition of new skills. The initial call for the ban was by the Brazilian Network for Humane Education (RedEH), an independent body comprising Brazilian professors and international collaborators dedicated to the implementation of replacement alternatives in education. The call was supported by InterNICHE, and many professors and other international organisations. The Brazilian Council of Veterinary Medicine, which is responsible for regulating the veterinary profession in Brazil, also stated its support for humane education and for the ban. The call was the first formal request, and it eventually led to the first legal resolution for the replacement of animal use in education in Brazil. This represents an important historic landmark in the advancement of science education.
A Proposal for a New Ethical Scheme Addressing the Use of Laboratory Animals for Biomedical Purposes
Authors: Delpire Véronique C. , Mepham T. Ben , Balls Michael
Published: April 2019

Current European Union legislation relating to the use of laboratory animals, such as Directive 86/609/EEC, was implemented before the full implications of transgenesis were recognised. An analysis of selected existing cost-benefit guidelines has shown that they do not provide for an adequate evaluation of the use of transgenic animals. In this paper, a proposal is made for a new ethical scheme, comprising a specific cluster of questions dealing with transgenic animals and the wider implications of their use. It is suggested that this scheme could be used at the European level in reviewing project proposals involving animal experimentation.
Transgenic Models for Prion Disease: Have They Outlived Their Useful Purpose?
Authors: Jenkins Elizabeth S. , Combes Robert D.
Published: April 2019

Prions are a recently identified class of proteinaceous pathological agents. Prion diseases are fatal neurological disorders, the importance of which is exemplified by the recent emergence of a novel variant of Creutzfeldt-Jacob disease (CJD) in humans. During pathogenesis, prion proteins undergo a conformational change, which converts the normal isoform to a pathogenic isoform. Several approaches are available for studying prion disease. The predominant approach has involved in vivo studies, especially involving transgenic mice. In vitro alternatives available for studying prion disease include a cell-free conversion assay, cell culture systems, and an immunoassay for the pathogenic form of the prion protein. Prion-like proteins have been identified in yeast, and therefore this constitutes another non-animal approach. Four main areas of prion research are discussed in this paper, to illustrate the potential applications and limitations of the in vivo and alternative systems. From this study, we conclude that, while current in vitro approaches can be used initially, in vivo studies are still needed to confirm data obtained in vitro. Priority should be given to the non-animal alternatives, as well as to developing new methods, and these should be given primary consideration at the outset of a project.
Bioethical Issues in the Generation and Use of Transgenic Farm Animals
Authors: Mepham T. Ben , Crilly Robert E.
Published: April 2019

The use of transgenic farm animals presents many challenges to the bioethicist, not least how to analyse the ethical issues within a framework that does not implicitly assume adherence to a normative ethical theory. One possible solution is to use a series of prima-facie principles applied to the interest groups affected by transgenesis. One such scheme is based on the four prima-facie principles of respect for non-maleficence, beneficence, autonomy and justice. This paper illustrates these with respect to transgenic farm animals. The aim of this is a systematic analysis, which includes positive and negative aspects, and which can therefore serve as a starting point for debate in which a range of views exist. One possible interpretation of this analysis is based on Three Rs concept. The use of transgenic farm animals appears to contradict this concept, because, although there is the potential for a reduction in animal numbers, at present, transgenesis is a rapidly expanding field, reversing the recent modest reductions in other areas of laboratory animal use. Moreover, transgenesis permits novel uses of farm animals, such as the production of proteins for human medicine where they were previously obtained from human blood (the opposite of replacement). The technique of transgenesis also misses the point as far as refinement is concerned, by refining the animal to suit the production or experimental protocol, instead of vice versa.
Welfare Assessment of Transgenic Animals: Behavioural Responses and Morphological Development of Newborn Mice
Authors: van der Meer Miriam , Costa Patrizia , Baumans Vera , Olivier Berend , van Zutphen Bert
Published: April 2019

Four groups of mice of the same inbred strain, but with different transgenic backgrounds (no treatment; integration of a functional corticotrophin-releasing factor [CRF] gene construct; integration of a non-functional CRF gene construct; transgenic technique without integration of a DNA construct) were compared, in order to identify and quantify indicators of discomfort in transgenic animals. This approach enables us to differentiate between the effects of the technique of transgenesis and the effects caused by the expression of the transgene. This paper emphasises the search for differences in the early post-natal development of the animals. To this end, newborn mice have been subjected to various behavioural tests; moreover, their growth and morphological characteristics were measured from birth up to the age of 3 weeks. The results indicate that the presence of the microinjected DNA-construct influences the survival rate during the first 2–3 days after birth. The average loss of pups was about 10%, in contrast to the groups without the DNA construct, in which none of the pups died. The increase in the relative body weight of pups with a functional CRF construct was significantly lower than in the other groups, but only during the first 11 days. No significant differences in morphological characteristics or behavioural development were observed between the four groups. This approach was found to be adequate for detecting a broad variety of behavioural and morphological characteristics. Before general conclusions about the extent to which the technique of transgenesis affects animal welfare can be drawn, more transgenic lines should be studied in this way.
Differential hypoxemia during venoarterial extracorporeal membrane oxygenation
Authors: Falk Lars , Sallisalmi Marko , Lindholm Jonas Andersson , Lindfors Mattias , Frenckner Björn , Broomé Michael , Broman Lars Mikael
Published: April 2019

Venoarterial extracorporeal membrane oxygenation, indicated for severe cardio-respiratory failure, may result in anatomic regional differences in oxygen saturation. This depends on cannulation, hemodynamic state, and severity of respiratory failure. Differential hypoxemia, often discrete, may cause clinical problems in peripheral femoro-femoral venoarterial extracorporeal membrane oxygenation, when the upper body is perfused with low saturated blood from the heart and the lower body with well-oxygenated extracorporeal membrane oxygenation blood. The key is to diagnose and manage fulminant differential hypoxemia, that is, a state that may develop where the upper body is deprived of oxygen. We summarize physiology, assessment of diagnosis, and management of fulminant differential hypoxemia during venoarterial extracorporeal membrane oxygenation. A possible solution is implantation of an additional jugular venous return cannula. In this article, we propose an even better solution, to drain the venous blood from the superior vena cava. Drainage from the superior vena cava provides superiority to venovenoarterial configuration in terms of physiological rationale, efficiency, safety, and simplicity in clinical circuit design.
Pressure and flow properties of cannulae for extracorporeal membrane oxygenation II: drainage (venous) cannulae
Authors: Broman Lars Mikael , Prahl Wittberg Lisa , Westlund C Jerker , Gilbers Martijn , Perry da Câmara Luisa , Westin Jan , Taccone Fabio Silvio , Malfertheiner Maximilian Valentin , Di Nardo Matteo , Swol Justyna , Vercaemst Leen , Barrett Nicholas A , Pappalardo Federico , Belohlavek Jan , Müller Thomas , Belliato Mirko , Lorusso Roberto
Published: April 2019

The use of extracorporeal life support devices such as extracorporeal membrane oxygenation in adults requires cannulation of the patient’s vessels with comparatively large diameter cannulae to allow circulation of large volumes of blood (>5 L/min). The cannula diameter and length are the major determinants for extracorporeal membrane oxygenation flow. Manufacturing companies present pressure-flow charts for the cannulae; however, these tests are performed with water. Aims of this study were 1. to investigate the specified pressure-flow charts obtained when using human blood as the circulating medium and 2. to support extracorporeal membrane oxygenation providers with pressure-flow data for correct choice of the cannula to reach an optimal flow with optimal hydrodynamic performance. Eighteen extracorporeal membrane oxygenation drainage cannulae, donated by the manufacturers (n = 6), were studied in a centrifugal pump driven mock loop. Pressure-flow properties and cannula features were described. The results showed that when blood with a hematocrit of 27% was used, the drainage pressure was consistently higher for a given flow (range 10%-350%) than when water was used (data from each respective manufacturer’s product information). It is concluded that the information provided by manufacturers in line with regulatory guidelines does not correspond to clinical performance and therefore may not provide the best guidance for clinicians.