bluuebunny/crossref_metadata_2025 huggingface dataset:
In Silico Toxicology in Drug Discovery — Concepts Based on Three-dimensional Models
Authors: Vedani Angelo , Smiesko Martin
Published: April 2019

Animal testing is still compulsory worldwide, for the approval of drugs and chemicals produced in large quantities. Computer-assisted ( in silico) technologies are considered to be efficient alternatives to in vivo experiments, and are therefore endorsed by many regulatory agencies, e.g. for use in the European REACH initiative. Advantages of in silico methods include: the possible study of hypothetical compounds; their low cost; and the fact that such virtual experiments are typically based on human data, thus making the question of interspecies transferability obsolete. Since the mid-1990s, computer-based technologies have become an indispensable tool in drug discovery — used primarily to identify small molecules displaying a stereospecific and selective binding to a regulatory macromolecule. Since toxic effects are still responsible for some 20% of the late-stage failures, there is a continuing need for in silico concepts which can be used to estimate a compound's ADMET ( adsorption, distribution, metabolism, elimination, toxicity) properties — in particular, toxicity. The aim of this paper is to provide an insight into computational technologies that allow for the prediction of toxic effects triggered by pharmaceuticals. As most adverse and toxic effects are mediated by unwanted interactions with macromolecules involved in biological regulatory systems, we have focused on methodologies that are based on three-dimensional models of small molecules binding to such entities, and discuss the results at the molecular level.
The In Chemico–In Silico Interface: Challenges for Integrating Experimental and Computational Chemistry to Identify Toxicity
Authors: Cronin Mark T.D. , Bajot Fania , Enoch Steven J. , Madden Judith C. , Roberts David W. , Schwöbel Johannes
Published: April 2019

A number of toxic effects are brought about by the covalent interaction between the toxicant and biological macromolecules. In chemico assays are available that attempt to identify reactive compounds. These approaches have been developed independently for pharmaceuticals and for other non-pharmaceutical compounds. The assays vary widely in terms of the macromolecule (typically a peptide) and the analytical technique utilised. For both sets of methods, there are great opportunities to capture in chemico information by using in silico methods to provide computational tools for screening purposes. In order to use these in chemico and in silico methods, integrated testing strategies are required for individual toxicity endpoints. The potential for the use of these approaches is described, and a number of recommendations to improve this extremely useful technique, in terms of implementing the Three Rs in toxicity testing, are presented.
The Current Status of Animal Use and Alternatives in Korean Veterinary Medical Schools
Authors: Lee Gwi Hyang , Choe Byung In , Kim Jin Suk , Hart Lynette A. , Han Jin Soo
Published: April 2019

Two new Korean laws regulating animal welfare and the humane use of animals in science came into effect in 2008 and 2009. Both these laws impose ethical committee review prior to the performance of animal experiments in research, testing and education. This study briefly summarises the new Korean laws, and investigates the current status regarding the numbers of animals used, the alternatives to animals which are used, the curricula relating to the humane use of animals, and ethical review practices in Korean veterinary education. Approximately 4,845 animals, representing 20 different species, were used in veterinary medical education in Korea in 2007. Korea has begun to introduce formal courses on animal welfare for the humane treatment of animals used in experiments, and an ethical protocol review system prior to animal use in education. Korea is moving toward better animal welfare, by incorporating practices consistent with international standards. The information presented represents the first such data gathered in Korea, which should prove useful for monitoring the implementation of replacement, reduction, and refinement measures in animal use for education purposes.
Validating Animal Models for Preclinical Research: A Scientific and Ethical Discussion
Authors: Varga Orsolya E. , Hansen Axel K. , Sandøe Peter , Olsson I. Anna S.
Published: April 2019

The use of animals to model humans in biomedical research relies on the notion that basic processes are sufficiently similar across species to allow extrapolation. Animal model validity is discussed in terms of the similarity between the model and the human condition it is intended to model, but no formal validation of models is applied. There is a stark contrast here with the use of non-animal alternatives in toxicology and safety studies, for which an extensive validation is required. We discuss both the potential and the limitations of validating preclinical animal models for proof-of-concept studies, by using an approach similar to that applied to alternative non-animal methods in toxicology and safety testing. A major challenge in devising a validation system for animal models is the lack of a clear gold standard with which to compare results. While a complete adoption of the validation approach for alternative methods is probably inappropriate for research animal models, key features, such as making data available for external validation and defining a strategy to run experiments in a way that permits meaningful retrospective analysis, remain highly relevant.
E-SovTox: An Online Database of the Main Publicly-available Sources of Toxicity Data concerning REACH-relevant Chemicals Published in the Russian Language
Authors: Sihtmäe Mariliis , Blinova Irina , Aruoja Villem , Dubourguier Henri-Charles , Legrand Nicolas , Kahru Anne
Published: April 2019

A new open-access online database, E-SovTox, is presented. E-SovTox provides toxicological data for substances relevant to the EU Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) system, from publicly-available Russian language data sources. The database contains information selected mainly from scientific journals published during the Soviet Union era. The main information source for this database — the journal, Gigiena Truda i Professional'nye Zabolevania [ Industrial Hygiene and Occupational Diseases], published between 1957 and 1992 — features acute, but also chronic, toxicity data for numerous industrial chemicals, e.g. for rats, mice, guinea-pigs and rabbits. The main goal of the above-mentioned toxicity studies was to derive the maximum allowable concentration limits for industrial chemicals in the occupational health settings of the former Soviet Union. Thus, articles featured in the database include mostly data on LD50 values, skin and eye irritation, skin sensitisation and cumulative properties. Currently, the E-SovTox database contains toxicity data selected from more than 500 papers covering more than 600 chemicals. The user is provided with the main toxicity information, as well as abstracts of these papers in Russian and in English (given as provided in the original publication). The search engine allows cross-searching of the database by the name or CAS number of the compound, and the author of the paper. The E-SovTox database can be used as a decision-support tool by researchers and regulators for the hazard assessment of chemical substances.
Replacing Animal Use in Physiology and Pharmacology Teaching in Selected Universities in Eastern Europe — Charting a Way Forward
Authors: Dewhurst David G. , Kojic Zvezdana Z.
Published: April 2019

The aims of this study were to explore the use of animals in teaching and the implementation of innovative technology-based teaching practices across a small sample of universities in Eastern Europe. The research methods used were a questionnaire circulated four weeks before a workshop took place (in October 2009, in Belgrade, Serbia), as well as focused, face-to-face group discussions, led by one of the authors during the workshop. Twenty-two faculty (physiologists and pharmacologists), from 13 Eastern European countries, attended the meeting. Fourteen of the eighteen schools represented at the workshop were making use of animals, in some instances in quite large numbers, for their teaching. For example, a single department at a Romanian university used over 250 animals per annum, and at least 1130 animals were used, per annum, across all of the institutions. The species used in largest numbers were the rat (34%), frog/toad (29%), mouse (22%), rabbit (10%), guinea-pig (4%) and dog (1%). None of the universities sampled had implemented institution-wide virtual learning environments (VLEs), although there were isolated instances of local use of VLEs. There was relatively little current use of technology-based teaching and learning resources, but there was considerable enthusiasm to modernise teaching and to introduce innovative learning and teaching methods. The major perceived barrier to the introduction of replacement alternatives was the lack of versions in local languages. There was a consensus view that developing local language exemplars and evaluating their usefulness was likely to have the greatest impact on animal use, at least in the short-term.
The Cell Transformation Assay: Toward a Statistical Classification of Mixed and Intermediate Foci Images
Authors: Procaccianti Claudio , Stefanini Federico M. , Urani Chiara
Published: April 2019

The human carcinogenicity evaluation of chemicals has a great impact on public health. In vitro methods, such as the cell transformation assay (CTA), allow for a fast and reliable assessment of the carcinogenic potential of a chemical compound in comparison with the standard two-year bioassay. The scoring and classification of foci in selected cell lines is performed, after staining, by light microscopy. Foci can be separated into three classes: type I, which are scored as non-transformed, and types II and III that are considered to include fully transformed foci. However, in a number of cases, even an expert is uncertain about the attribution of a focus to a given class, due to its mixed or intermediate nature. Here, we suggest a simple approach to classifying mixed or intermediate foci by exploiting the quantitative information available from images, which is captured by statistical descriptors. A quantitative index is proposed, to describe the degree of dissimilarity of mixed and intermediate images to the three well-distinguished classes.
Implementation of the Three Rs in the Human Hazard Assessment of Brazilian Medicinal Plants: An Evaluation of the Cytotoxic and Genotoxic Potentials of Dipteryx alata Vogel
Authors: Esteves-Pedro Natália M. , Rodas Andrea C.D. , Dal Belo Cháriston A. , Oshima-Franco Yoko , Santos Márcio G. dos , Lopes Patricia S.
Published: April 2019

In Brazil, medicinal plants are widely used by the indigenous people, which leads to a constant requirement for toxicity tests to be performed on the plant extracts. Although the current Brazilian Directive 90/2004 on the preclinical toxicity testing of phytotherapeutics recommends only in vivo tests, some Brazilian researchers would like to change this situation by implementing the Three Rs in the toxicological testing of medicinal plants. The present study evaluated the cytotoxic and genotoxic potentials of bark extracts from Dipteryx alata Vogel, a medicinal plant of the Brazilian cerrado, by using CHO-K1 (Chinese hamster ovary) cells. An IC50 value was obtained, which corresponded to 0.16mg/ml of plant extract, and from this the equivalent LD50 was determined as 705mg/kg. In order to determine the genotoxic potential of the sample, the frequency of micronucleus formation was assessed. CHO-K1 cells were exposed, during targeted mitosis, to different concentrations of plant extract and cytochalasin B, in the presence and absence of an appropriate metabolic activation system (an S9 mix). The results obtained indicated that it might be possible to implement the Three Rs in assessing the potential human hazard of medicinal plants. The publication of such data can increase awareness of the Three Rs by showing how to optimise the management of animal use, if in vivo toxicological experiments are required.
Veterinary Science Student Preferences for the Source of Dog Cadavers Used in Anatomy Teaching
Authors: Tiplady Catherine , Lloyd Shan , Morton John
Published: April 2019

Live animals and cadavers are integral to veterinary education. In the year of this survey (2008), and in at least the five preceding years, cadavers obtained by euthanasia of healthy pound dogs and ex-racing greyhounds were dissected by students, during their veterinary anatomy classes at the University of Queensland School of Veterinary Science. Students may have ethical concerns about this. An alternative approach was to use donated dog cadavers. These are owned pet dogs that have died of natural causes or have been euthanised for medical reasons, and have been donated by their owners for the purposes of veterinary education. Veterinary students at the School were surveyed in 2008, in order to determine their preferences for cadaver source. Data from 406 questionnaires were analysed. Third-year and fifth-year veterinary students were more likely than first-year students to prefer pound-dog/greyhound cadavers over donated cadavers for anatomy dissection ( p ≤ 0.002). Between 32% and 45% of the students had no preference for either source of cadaver. These findings are consistent with the hypothesis that veterinary students become more accepting of the euthanasia of unwanted healthy animals for education as they progress through the veterinary programme, in contexts such as the current study. This could occur due to increased acceptance of the euthanasia of healthy animals generally, a decline in moral development, desensitisation, and/or the belief that healthy animal cadavers offer a superior learning experience.
The In Vitro Use of the Hair Follicle Closure Technique to Study the Follicular and Percutaneous Permeation of Topically Applied Drugs
Authors: Stahl Jessica , Niedorf Frank , Wohlert Mareike , Kietzmann Manfred
Published: April 2019

Recent studies on follicular permeation emphasise the importance of hair follicles as diffusion pathways, but only a limited amount of data are available about the follicular permeation of topically applied drugs. This study examines the use of a hair follicle closure technique in vitro, to determine the participation of hair follicles in transdermal drug penetration. Various substances, with different lipophilicities, were tested: caffeine, diclofenac, flufenamic acid, ibuprofen, paracetamol, salicylic acid and testosterone. Diffusion experiments were conducted with porcine skin, the most common replacement material for human skin, in Franz-type diffusion cells over 28 hours. Different experimental settings allowed the differentiation between interfollicular and follicular permeation after topical application of the test compounds. A comparison of the apparent permeability coefficients of the drugs demonstrates that the percutaneous permeations of caffeine and flufenamic acid were significantly higher along the hair follicles. In the cases of paracetamol and testosterone, the follicular pathway appears to be of importance, while no difference was found between interfollicular and follicular permeation for diclofenac, ibuprofen and salicylic acid. Thus, the hair follicle closure technique represents an adequate in vitro method for gaining information about follicular or percutaneous permeation, and can replace in vivo testing in animals or humans.
On Challenging the Prevailing Dogma: Nobel Prizes and Pink Diamonds
Authors: Balls Michael
Published: April 2019

Ones superiors in the hierarchy are not always right
Automated Homecage Behavioural Analysis and the Implementation of the Three Rs in Research Involving Mice
Authors: Richardson Claire A.
Published: April 2019

Automated homecage behavioural analysis systems have the potential to both refine scientific procedures and reduce the numbers of mice used
A Vision Becoming Reality
Authors: Langley Gill
Published: April 2019

Non-animal science in toxicology and health research has been progressing for decades, but only now is it being seen widely as advanced science. The emergence of novel human biology-based tools and models, combined with legislative and regulatory change, a 21st century concept for toxicology, continuing failures in the drug pipeline, and systematic critiques of animal models, have created a pivotal moment of change. The leading edge is starting to become the norm. Humans and other animals are likely to benefit as a result.
Humane Society International's Global Campaign to End Animal Testing
Authors: Seidle Troy
Published: April 2019

The Research & Toxicology Department of Humane Society International (HSI) operates a multifaceted and science-driven global programme aimed at ending the use of animals in toxicity testing and research. The key strategic objectives include: a) ending cosmetics animal testing worldwide, via the multinational Be Cruelty-Free campaign; b) achieving near-term reductions in animal testing requirements through revision of product sector regulations; and c) advancing humane science by exposing failing animal models of human disease and shifting science funding toward human biology-based research and testing tools fit for the 21st century. HSI was instrumental in ensuring the implementation of the March 2013 European sales ban for newly animal-tested cosmetics, in achieving the June 2013 cosmetics animal testing ban in India as well as major cosmetics regulatory policy shifts in China and South Korea, and in securing precedent-setting reductions in in vivo data requirements for pesticides in the EU through the revision of biocides and plant protection product regulations, among others. HSI is currently working to export these life-saving measures to more than a dozen industrial and emerging economies.
Evaluation of the Cytotoxic Effects of Humid Lightweight Coal Ash derived from the Disposal of Waste on Normal Human Keratinocyte and Endothelial Cell Lines in 2-D and 3-D Culture
Authors: Scanarotti Chiara , Vernazza Stefania , Brignone Massimiliano , Danailova Jenia , Pronzato Maria A. , Bassi Anna M.
Published: April 2019

The presence of waste in the environment has frequently been indicated as a significant risk to human health. Therefore, landfill sites and the disposal of urban solid and non-hazardous waste by incineration are subject to much environmental monitoring, in addition to the regulations already in place. However, little action has been taken, and consequently no specific legislation exists, in relation to the assessment of the real biological risk of various substances, including chemical mixtures and ashes, derived from the incineration processes. This study assessed the cytotoxic potential of humid lightweight coal ash (LA) derived from incineration processes and waste management, on two cell lines: NCTC 2544 normal human keratinocytes and HECV endothelial cells. To reach this goal and to assess more-realistic methods for animal replacement, we employed different in vitro experimental approaches: acute and longer exposure to LA, by direct and indirect contact (0–2mg/ml and 16mg, respectively), both in 2-D and 3-D cultures. In 2-D HECV cultures, we observed a decrease in the viability index, but only during direct contact with LA doses higher than 0.1mg/ml. Moreover, some striking differences in cytotoxicity were observed between the 2-D and 3-D models. Taken together, these observations indicate that, for studying pollutant toxicity during longer exposure times, 3-D cultures in direct contact with the pollutant seem to offer a more suitable approach — they mimic the in vivo behaviour of cells more realistically and under strictly controlled conditions. Thus, in readiness for possible forthcoming European regulations, we believe that the proposed study, even in its preliminary phase, can provide new advice on the assessment of the toxic and biological potential of particular chemical compounds derived from waste management processes.
Alternatives to Animal Bioassays for Prions
Authors: Beekes Michael , Wagenführ Katja
Published: April 2019

Combined PMCA and cell assays are being optimised for use as an effective, or even superior, alternative to quantitative prion bioassays in laboratory rodents
The QSPR-THESAURUS: The Online Platform of the CADASTER Project
Authors: Brandmaier Stefan , Peijnenburg Willie , Durjava Mojca K. , Kolar Boris , Gramatica Paola , Papa Ester , Bhhatarai Barun , Kovarich Simona , Cassani Stefano , Roy Partha Pratim , Rahmberg Magnus , Öberg Tomas , Jeliazkova Nina , Golsteijn Laura , Comber Mike , Charochkina Larisa , Novotarskyi Sergii , Sushko Iurii , Abdelaziz Ahmed , D'Onofrio Elisa , Kunwar Prakash , Ruggiu Fiorella , Tetko Igor V.
Published: April 2019

The aim of the CADASTER project (CAse Studies on the Development and Application of in Silico Techniques for Environmental Hazard and Risk Assessment) was to exemplify REACH-related hazard assessments for four classes of chemical compound, namely, polybrominated diphenylethers, per and polyfluorinated compounds, (benzo)triazoles, and musks and fragrances. The QSPR-THESAURUS website ( http://qspr-thesaurus.eu ) was established as the project's online platform to upload, store, apply, and also create, models within the project. We overview the main features of the website, such as model upload, experimental design and hazard assessment to support risk assessment, and integration with other web tools, all of which are essential parts of the QSPR-THESAURUS.
Estimation of the Toxicity of Silver Nanoparticles by Using Planarian Flatworms
Authors: Kustov Leonid , Tiras Kharlampii , Al-Abed Souhail , Golovina Natalia , Ananyan Mikhail
Published: April 2019

The regeneration of planarian flatworms — specifically, changes to the area of the regeneration bud (blastema) after surgical dissection — was proposed for use as a robust tool for estimating the toxicity of silver nanoparticles. The use of Planaria species, due to their unique regenerative capacity, could result in a reduction in the use of more-traditional laboratory animals for toxicity testing. With our novel approach, silver nanoparticles were found to be moderately toxic to the planarian, Girardia tigrina.
Quantitative Structure–Sorption Relationships of Pesticides Used in the Sugarcane Industry in the Northern Coastal Area of Paraíba State, Brazil
Authors: Soares Gabriela C. da S. , Silva Luana de M. e , Farias Carlos H. de A. , Scotti Luciana , Scotti Marcus T.
Published: April 2019

Sorption coefficients (Koc) are useful in the prediction of whether a pesticide will remain dissolved in solution or will become adsorbed onto soil particles after its application. Measuring this process experimentally is difficult, expensive and time-consuming. Hence, much effort has been directed toward estimating Koc through statistical modelling. In this study, we investigated the physicochemical properties of pesticides employed by a local sugarcane company, in the northern coastal plain of Paraíba state in Brazil, by using several molecular descriptors, among them, GRid INdependent Descriptors (GRIND). Quantitative assessment of the structure–property relationship (QSPR) model indicated that size, shape, octanol–water coefficient, solubility and the balance between hydrophilic and lipophilic regions, are all relevant to Koc values.
9. The Toxicity Testing Problem
Authors: Balls Michael
Published: April 2019

The field of toxicity testing is an urgent humanitarian problem, for it regularly involves a finite and large incidence of distress which is often considerable and sometimes acute
The Three Rs — Opportunities for Improving Animal Welfare and the Quality of Scientific Research
Authors: Combes Robert D. , Balls Michael
Published: April 2019

In 2013, an undercover investigation by the BUAV raised serious concerns about the use, treatment and care of laboratory animals involved in regulated procedures at Imperial College, London. This led to an inquiry, set up by the college, which found deficiencies in the local ethical review process and a general lack of focus on the implementation of the Three Rs ( Replacement, Refinement and Reduction). The Three Rs concept is the foundation of UK and EU legislation, but surveys of the published literature show that lack of its adoption is widespread. In spite of numerous guidelines, publications and publicity material extolling the benefits of the Three Rs to both animals and science, as well as substantial advances in the development, validation, and deployment of mechanistically-based non-animal methods, many scientists prefer to use traditional animal-based approaches. In addition, such scientists tend to pay less attention than they should to strategic planning, experimental design and the choice of appropriate statistical procedures. They are often unaware of the existence of replacement test methods to address all or some of their objectives, and are reluctant to develop and use new replacement methods. We explore some possible reasons for these shortcomings. We summarise the welfare and scientific effects of each of the Three Rs, and argue that: a) there is an urgent need for evidence to be made readily accessible to prospective licensees, which directly demonstrates the beneficial effects on animal welfare of the implementation of the Three Rs, separately and in combination, and the direct link this has with the quality of the scientific data obtained; b) a detailed systematic review of this evidence should be undertaken to augment the inadequate content of the prescribed Module 5 licensee training offered currently in the UK; c) such training (including that suggested in new EU-wide proposals) should be much more comprehensive, with stronger emphasis on the Three Rs, all parts of the syllabus should be fully examined, and there should be no exemptions from Module 5 training; and d) as the responsible Government department in the UK, the Home Office should take measures to tighten up its guidance for local ethical review, and its system of inspection of designated establishments, to obviate the justification for future undercover investigations.
Personal Reflections on Veterinary Science Training and the Three Rs
Authors: Elliott Rosemary
Published: April 2019

Students against the thoughtless use of animals in veterinary education need support to be true to their values, to use science to uphold them, and to never give up on advocating for the highest standards of animal welfare
Lost in Translation: The Need for Better Tools
Authors: Penco Susanna , Venco Elena , Lio Alfredo
Published: April 2019

Although for most pharmaceutical compounds the final aim is improving human health, almost all the methods used to identify and pursue therapeutic targets and to obtain new potential drugs have traditionally focused on animal models
2013 Lush Science Prize
Authors: Harrison Rob
Published: April 2019

The annual Lush Science Prize is designed to reward outstanding contributions to 21st Century Toxicology Research. A Background Paper is prepared each year prior to the judging process, in order to provide the judging panel with a brief overview of current developments in the field of Replacement alternatives, particularly those relevant to the concept of toxicity pathways. The Background Paper includes information on some key institutional developments in the area — such as the OECD's Adverse Outcome Pathway Project, the Hamner Institute's work, and the Human Toxome Project, and on the phenomenon of collaborative computer systems relevant to the field. From the literature review that was also performed as part of the background research, the two papers receiving the highest score were recommended for consideration by the judges for the 2013 Science Prize.
The EpiOcular™ Eye Irritation Test is the Method of Choice for the In Vitro Eye Irritation Testing of Agrochemical Formulations: Correlation Analysis of EpiOcular Eye Irritation Test and BCOP Test Data According to the UN GHS, US EPA and Brazil ANVISA Classification Schemes
Authors: Kolle Susanne N. , Moreno Maria Cecilia Rey , Mayer Winfried , van Cott Andrew , van Ravenzwaay Bennard , Landsiedel Robert
Published: April 2019

The Bovine Corneal Opacity and Permeability (BCOP) test is commonly used for the identification of severe ocular irritants (GHS Category 1), but it is not recommended for the identification of ocular irritants (GHS Category 2). The incorporation of human reconstructed tissue model-based tests into a tiered test strategy to identify ocular non-irritants and replace the Draize rabbit eye irritation test has been suggested (OECD TG 405). The value of the EpiOcular™ Eye Irritation Test (EIT) for the prediction of ocular non-irritants (GHS No Category) has been demonstrated, and an OECD Test Guideline (TG) was drafted in 2014. The purpose of this study was to evaluate whether the BCOP test, in conjunction with corneal histopathology (as suggested for the evaluation of the depth of the injury) and/or the EpiOcular-EIT, could be used to predict the eye irritation potential of agrochemical formulations according to the UN GHS, US EPA and Brazil ANVISA classification schemes. We have assessed opacity, permeability and histopathology in the BCOP assay, and relative tissue viability in the EpiOcular-EIT, for 97 agrochemical formulations with available in vivo eye irritation data. By using the OECD TG 437 protocol for liquids, the BCOP test did not result in sufficient correct predictions of severe ocular irritants for any of the three classification schemes. The lack of sensitivity could be improved somewhat by the inclusion of corneal histopathology, but the relative viability in the EpiOcular-EIT clearly outperformed the BCOP test for all three classification schemes. The predictive capacity of the EpiOcular-EIT for ocular non-irritants (UN GHS No Category) for the 97 agrochemical formulations tested (91% sensitivity, 72% specificity and 82% accuracy for UN GHS classification) was comparable to that obtained in the formal validation exercise underlying the OECD draft TG. We therefore conclude that the EpiOcular-EIT is currently the best in vitro method for the prediction of the eye irritation potential of liquid agrochemical formulations.
17. The Choice of Procedures
Authors: Balls Michael
Published: April 2019

The scientifically-justifiable choice of procedure is a crucial issue in animal experimentation, in the interests of both humanity and efficiency
Barriers to the Uptake of Human-based Test Methods, and how to Overcome Them
Authors: Archibald Kathy , Drake Tamara , Coleman Robert
Published: April 2019

Although there is growing concern as to the questionable value of animal-based methods for determining the safety and efficacy of new medicines, which has in turn led to many groups developing innovative human-based methods, there are many barriers to their adoption for regulatory submissions. The reasons for this are various, and include a lack of confidence that the available human-based methods, be they in vivo, in silico or in vitro, can be sufficiently predictive of clinical outcomes. However, this is not the only problem: the issue of validation presents a serious impediment to progress, a particularly frustrating situation, in view of the fact that the existing animal-based methods have never themselves been formally validated. Superimposed upon this is the issue of regulatory requirements, where, although regulators may be willing to accept non-animal approaches in place of particular animal tests, nowhere is this explicitly stated in their guidelines. Such problems are far from trivial, and represent major hurdles to be overcome. In addition, there are a range of other barriers, real or self-imposed, that are hindering a more-predictive approach to establishing a new drug's clinical safety and efficacy profiles. Some of these barriers are identified, and ways forward are suggested.
Toward the Replacement of Animal Experiments through the Bioinformatics-driven Analysis of ‘Omics’ Data from Human Cell Cultures
Authors: Grafström Roland C. , Nymark Penny , Hongisto Vesa , Spjuth Ola , Ceder Rebecca , Willighagen Egon , Hardy Barry , Kaski Samuel , Kohonen Pekka
Published: April 2019

This paper outlines the work for which Roland Grafström and Pekka Kohonen were awarded the 2014 Lush Science Prize. The research activities of the Grafström laboratory have, for many years, covered cancer biology studies, as well as the development and application of toxicity-predictive in vitro models to determine chemical safety. Through the integration of in silico analyses of diverse types of genomics data (transcriptomic and proteomic), their efforts have proved to fit well into the recently-developed Adverse Outcome Pathway paradigm. Genomics analysis within state-of-the-art cancer biology research and Toxicology in the 21st Century concepts share many technological tools. A key category within the Three Rs paradigm is the Replacement of animals in toxicity testing with alternative methods, such as bioinformatics-driven analyses of data obtained from human cell cultures exposed to diverse toxicants. This work was recently expanded within the pan-European SEURAT-1 project (Safety Evaluation Ultimately Replacing Animal Testing), to replace repeat-dose toxicity testing with data-rich analyses of sophisticated cell culture models. The aims and objectives of the SEURAT project have been to guide the application, analysis, interpretation and storage of ‘omics’ technology-derived data within the service-oriented sub-project, ToxBank. Particularly addressing the Lush Science Prize focus on the relevance of toxicity pathways, a ‘data warehouse’ that is under continuous expansion, coupled with the development of novel data storage and management methods for toxicology, serve to address data integration across multiple ‘omics’ technologies. The prize winners’ guiding principles and concepts for modern knowledge management of toxicological data are summarised. The translation of basic discovery results ranged from chemical-testing and material-testing data, to information relevant to human health and environmental safety.
The Use of a Decision Tree Based on the Rabies Diagnosis Scenario, to Assist the Implementation of Alternatives to Laboratory Animals
Authors: Bones Vanessa Carli , Molento Carla Forte Maiolino
Published: April 2019

Brazilian federal legislation makes the use of alternatives mandatory, when there are validated methods to replace the use of laboratory animals. The objective of this paper is to introduce a novel decision tree (DT)-based approach, which can be used to assist the replacement of laboratory animal procedures in Brazil. This project is based on a previous analysis of the rabies diagnosis scenario, in which we identified certain barriers that hinder replacement, such as: a) the perceived higher costs of alternative methods; b) the availability of staff qualified in these methods; c) resistance to change by laboratory staff; d) regulatory obstacles, including incompatibilities between the Federal Environmental Crimes Act and specific norms and working practices relating to the use of laboratory animals; and e) the lack of government incentives. The DT represents a highly promising means to overcome these reported barriers to the replacement of laboratory animal use in Brazil. It provides guidance to address the main obstacles, and, followed step-by-step, would lead to the implementation of validated alternative methods (VAMs), or their development when such alternatives do not exist. The DT appears suitable for application to laboratory animal use scenarios where alternative methods already exist, such as in the case of rabies diagnosis, and could contribute to increase compliance with the Three Rs principles in science and with the current legal requirements in Brazil.
Transparency and Public Involvement in Animal Research
Authors: Pound Pandora , Blaug Ricardo
Published: April 2019

To be legitimate, research needs to be ethical, methodologically sound, of sufficient value to justify public expenditure and be transparent. Animal research has always been contested on ethical grounds, but there is now mounting evidence of poor scientific method, and growing doubts about its clinical value. So what of transparency? Here we examine the increasing focus on openness within animal research in the UK, analysing recent developments within the Home Office and within the main group representing the interests of the sector, Understanding Animal Research. We argue that, while important steps are being taken toward greater transparency, the legitimacy of animal research continues to be undermined by selective openness. We propose that openness could be increased through public involvement, and that this would bring about much needed improvements in animal research, as it has done in clinical research.
In Vitro Assessment of Skin Irritation Potential of Surfactant-based Formulations by Using a 3-D Skin Reconstructed Tissue Model and Cytokine Response
Authors: Walters Russel M. , Gandolfi Lisa , Mack M. Catherine , Fevola Michael , Martin Katharine , Hamilton Mathew T. , Hilberer Allison , Barnes Nicole , Wilt Nathan , Nash Jennifer R. , Raabe Hans A. , Costin Gertrude-Emilia
Published: April 2019

The personal care industry is focused on developing safe, more efficacious, and increasingly milder products, that are routinely undergoing preclinical and clinical testing before becoming available for consumer use on skin. In vitro systems based on skin reconstructed equivalents are now established for the preclinical assessment of product irritation potential and as alternative testing methods to the classic Draize rabbit skin irritation test. We have used the 3-D EpiDerm™ model system to evaluate tissue viability and primary cytokine interleukin-1α release as a way to evaluate the potential dermal irritation of 224 non-ionic, amphoteric and/or anionic surfactant-containing formulations, or individual raw materials. As part of our testing programme, two representative benchmark materials with known clinical skin irritation potential were qualified through repeated testing, for use as references for the skin irritation evaluation of formulations containing new surfactant ingredients. We have established a correlation between the in vitro screening approach and clinical testing, and are continually expanding our database to enhance this correlation. This testing programme integrates the efforts of global manufacturers of personal care products that focus on the development of increasingly milder formulations to be applied to the skin, without the use of animal testing.
The Use of Animals in Medical Research — A Historical Perspective
Authors: Zwolińska Jolanta
Published: April 2019

Even though not human, animals are not research equipment
A Preliminary Comparison of LiverBeads™ with a Conventional Rat Hepatocyte Culture Preparation: Some Aspects of Xenobiotic Metabolism and Related Toxicity
Authors: Hammond Alison H. , Garle Michael J. , Fry Jeffrey R.
Published: April 2019

LiverBeads™ are made of hepatocytes that are immobilised and cryopreserved in alginate gel. They have great potential as an easily transportable and easily handled source of hepatocytes for use in in vitro pharmacotoxicology. In this study, we compared the drug metabolising capacity of LiverBeads in culture with that of conventional cultures and of cultures derived from cryopreserved cells. Trypan blue exclusion, lactate dehydrogenase and DNA content were measured in LiverBead cultures. The levels were all similar to those of the conventional cultures, as were the toxicities of precocene II and allyl alcohol, although more variability was seen in the LiverBeads than in the conventional cultures. The cytochrome P450-dependent activity 3,4-dimethyl-7-ethoxycoumarin -O-dealkylase, was reduced in the LiverBeads when compared to the conventional cultures, although the pattern of conjugation was very similar. In addition, the inducibility of cytochrome P4504A was demonstrated in LiverBeads. Cultures from cryopreserved cells were more susceptible to the toxicants tested, and contained less lactate dehydrogenase and DNA than the conventional cultures. Overall, in terms of the aspects of drug metabolism measured, the cultures from LiverBeads appeared to be equivalent to conventional cultures, and superior to cultures from cryopreserved cells.
Human Bone Marrow Cell Culture: A Sensitive Method for Determination of the Biocompatibility of Implant Materials
Authors: Wilke Axel , Landgraff Maximilian , Orth Joachim , Poenitz Holger , Kienapfel Heino , Boelte Kerstin , Griss Peter , Franke Rolf P.
Published: April 2019

The objective of this study was to develop a test method for determining the cytotoxicity and biocompatibility of various biomaterials that are used in orthopaedic surgery. This method is based on the use of a human bone marrow cell culture and was developed as an alternative to animal experiments. Human bone marrow cell culture has certain advantages over other cell culture models, as its results show a greater conformity with animal experimental results and clinical studies. Primary cell adherence, cell number, cell proliferation, production of extracellular matrix, cell viability and cell differentiation were used as indicative parameters of biocompatibility. After 2 weeks in culture, differences could be observed between the biomaterials with respect to these parameters. Cell numbers were greatest on the hydroxyapatite ceramic specimens, but were decreased on the titanium alloy specimens. Extracellular matrix hydroxyapatite production was high for ceramics, but reduced for titanium specimens. The polymers allowed only a few cells to adhere, and there were no signs of extracellular matrix production. The influence of biomaterials on differentiation of large numbers of cells was analysed by using flow cytophotometry. There were similar populations of T cells and monocytes on all specimens. However, extended B cell and granulocyte populations were observed with titanium and polyethylene.
The use of Long-term Hepatocyte Cultures for Detecting Induction of Drug Metabolising Enzymes: The Current Status
Authors: Coecke Sandra , Rogiers Vera , Bayliss Martin , Castell José , Doehmer Johannes , Fabre Gérard , Fry Jeffrey , Kern Armin , Westmoreland Carl
Published: April 2019

In this report, metabolically competent in vitro systems have been reviewed, in the context of drug metabolising enzyme induction. Based on the experience of the scientists involved, a thorough survey of the literature on metabolically competent long-term culture models was performed. Following this, a prevalidation proposal for the use of the collagen gel sandwich hepatocyte culture system for drug metabolising enzyme induction was designed, focusing on the induction of the cytochrome P450 enzymes as the principal enzymes of interest. The ultimate goal of this prevalidation proposal is to provide industry and academia with a metabolically competent in vitro alternative for long-term studies. In an initial phase, the prevalidation study will be limited to the investigation of induction. However, proposals for other long-term applications of these systems should be forwarded to the European Centre for the Validation of Alternative Methods for consideration. The prevalidation proposal deals with several issues, including: a) species; b) practical prevalidation methodology; c) enzyme inducers; and d) advantages of working with independent expert laboratories. Since it is preferable to include other alternative tests for drug metabolising enzyme induction, when such tests arise, it is recommended that they meet the same level of development as for the collagen gel sandwich long-term hepatocyte system. Those tests which do so should begin the prevalidation and validation process.
An Interlaboratory Validation Study of the Improved Transformation Assay Employing Balb/c 3T3 Cells: Results of a Collaborative Study on the Two-stage Cell Transformation Assay by the Non-genotoxic Carcinogen Study Group
Authors: Tsuchiya Toshiyuki , Umeda Makoto , Nishiyama Hiroshi , Yoshimura Isao , Ajimi Shozo , Asakura Masumi , Baba Hiroshi , Dewa Yasuaki , Ebe Youji , Fushiwaki Yuichi , Hamada Shuichi , Hamamura Tetsuo , Hayashi Makoto , Iwase Yumiko , Kajiwara Yoshitsugu , Kasahara Yasushi , Kawabata Masayoshi , Kitada Emiko , Kubo Kinya , Mashiko Kaori , Miura Daisaku , Mizuhashi Fukutaro , Mizuno Fumio , Nakajima Madoka , Nakamura Yoshiyuki , Nobe Naoko , Oishi Hidetoshi , Ota Erina , Sakai Ayako , Sato Miho , Shimada Sawako , Sugiyama Toshie , Takahashi Chitose , Takeda Yuko , Tanaka Noriho , Toyoizumi Chikako , Tsutsui Takeki , Wakuri Shinobu , Yajima Satoshi , Yajima Nobuhiro
Published: April 2019

The Non-genotoxic Carcinogen Study Group of the Environmental Mutagen Society of Japan organised the first step of an interlaboratory validation study on an improved cell transformation assay employing Balb/c 3T3 A31-1-1 cells. Nineteen laboratories participated in this study. The modified transformation assay was evaluated for its responsiveness, its inter-laboratory reproducibility and its transferability. In this study, a mixture of Dulbecco's modified Eagle's medium and nutrient mixture F12, supplemented with insulin–transferrin– ethanolamine–sodium selenite and 2% fetal bovine serum (FBS) was used during the period of expression of transformed foci, intead of the usual minimum essential medium with 10% FBS. 20-Methylcholanthrene (MCA) and 12 -O-tetradecanoylphorbol-13-acetate (TPA) were selected as a prototype initiator and a tumour promoter, respectively. Two series of experiments were conducted. In the first series, the transformation activity of MCA was examined at various concentrations. In the absence of the promoting treatment with TPA, exposure to MCA only weakly induced transformed foci. In the presence of 0.1μg/ml TPA, all laboratories observed significant dose-dependent increases in the number of transformed foci with increasing MCA concentrations. In the second series of experiments, various concentrations of TPA were tested. In the absence of initiating treatment with MCA, exposure to TPA weakly induced transformed foci in about half of the laboratories. In the presence of 0.2μg/ml MCA, all the laboratories observed significant dose-dependent increases in the number of transformed foci with increasing TPA concentrations. The results from this study support the usefulness of this modified two-stage transformation assay with Balb/c 3T3 cells.
Managing health records in the Bulawayo and Matabeleland South provinces hospitals, Zimbabwe
Authors: Masuku Mehluli , Ngulube Patrick
Published: April 2019

The purpose of this study was to investigate the management of health records in the Bulawayo and Matabeleland South provinces in Zimbabwe. The objectives of the study were to understand how health records were being managed in the two hospitals, to establish the availability of, and analyse standard procedures and guidelines that informed both the National Archives of Zimbabwe (NAZ) and hospitals in managing health records. The study also sought to establish the level of professional training for health records management staff in hospitals under study. A qualitative case study design was employed and data was gathered through questionnaires, interviews, observation and document analysis. Data was analysed thematically based on the objectives of the study. The study revealed that the strategies for the management of health records in hospitals were inadequate. There were no documented health records management standards to guide the management of these records in hospitals. It was also established that the majority of health records personnel in hospitals did not possess records management qualifications. As a recommendation, the hospitals and NAZ should draft and implement health records management standards to provide guidance on the management of health records. Hospitals should employ staff with the requisite records management qualifications and offer them continuous training.