LATEST ISSUE

Volume 42, Issue 4 – August 2014

PCR amplification of CTGF

 

Editorial: Openness on Animal Research: The Gauntlet has been Thrown Down

Michael Balls

The delivery of the UK Government’s and Concordat’s commitments to greater openness on animal research is eagerly awaited. Meanwhile, the questions raised by two studies on the use of animal tests to predict the toxic effects of drugs in humans should be answered. Procedures applied to protected laboratory animals, which may cause them pain, suffering, distress and or lasting harm, are only morally acceptable, and should only be legally permissible, if they are scientifically justifiable.
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News & Views

ATLA Staff Writer

Nanoparticle Use in Imaging
Effects of Alcohol on the Brain
An In Vitro BBB Model
Acute Kidney Injury Studied In Vitro
Gut Microbes Affected by Gender–Diet Interactions
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CAAT News & Views

CAAT Writer

CAAT Receives Award for Raising Standards for Animal Welfare
CAAT Agency Partners Give Input for Future CAAT Programmes
Green Toxicology: Application of Predictive Toxicology
Job Opening: CEO of CAAT–Europe
Call for Proposals: 2015 Science-based Refinement Awards
Recent Publications
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Corrigendum ATLA 42, 31–51, 2014

Matteo Cassotti, Davide Ballabio, Viviana Consonni, Andrea Mauri, Igor V. Tetko and Roberto
Todeschini

Prediction of Acute Aquatic Toxicity Toward Daphnia magna by using the GA-kNN Method
On page 37, the depiction of experimental versus predicted responses for the training set shown in Figure 2a is inaccurate. The error was noticed after the data were re-analysed and a miscalculation in the indexing of the molecules was found.

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Student Perspectives on the Use of Alternative Methods for Teaching in Veterinary Faculties

Magda Sachana, Alexandros Theodoridis, Cristina Cortinovis, Fabiola Pizzo, Evaggelos Kehagias, Marco Albonico and Francesca Caloni

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.

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Lung Fibrosis: Drug Screening and Disease Biomarker Identification with a Lung Slice Culture Model and Subtracted cDNA Library

Tong Guo, Ka Yee Lok, Changhe Yu and Zhuo Li

Pulmonary fibrosis is a progressive and irreversible disorder with no appropriate cure. A
practical and effective experimental model that recapitulates the disease will greatly benefit the research community and, ultimately, patients. In this study, we tested the lung slice culture (LSC) system for its potential use in drug screening and disease biomarker identification. Fibrosis was induced by treating rat lung slices with 1ng/ml TGF-β1 and 2.5μM CdCl2, quantified by measuring the content of hydroxyproline, and confirmed by detecting the expression of collagen type III alpha 1 (Col3α1) and connective tissue growth factor (CTGF) genes. The anti-fibrotic effects of pirfenidone, spironolactone and eplerenone were assessed by their capability to reduce hydroxyproline content. A subtractive hybridisation technique was used to create two cDNA libraries (subtracted and unsubtracted) from lung slices. The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was employed to assess the subtraction efficiency of the subtracted cDNA library. Clones from the two libraries were sequenced and the genes were identified by performing a BLAST search on the NCBI GenBank database. Furthermore, the relevance of the genes to fibrosis formation was verified. The results presented here show that fibrosis was effectively induced in cultured lung slices, which exhibited significantly elevated levels of hydroxyproline and Col3α1/CTGF gene expression. Several inhibitors have demonstrated their anti-fibrotic effects by significantly reducing hydroxyproline content. The subtracted cDNA library, which was enriched for differentially expressed genes, was used to successfully identify genes associated with fibrosis. Collectively, the results indicate that our LSC system is an effective model for the screening of drug candidates and for disease biomarker identification.

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The Three Rs — Opportunities for Improving Animal Welfare and the Quality of Scientific Research

Robert D. Combes and Michael Balls

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.

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Statistics of Scientific Procedures on Living Animals 2013:Experimentation Continues to Rise

Michelle Hudson-Shore

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.

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A ‘Road Map’ Toward Ending Severe Suffering of Animals Used in Research and Testing

Elliot Lilley, Penny Hawkins and Maggy Jennings

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.
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FRAME is the Fund for the Replacement of Animals in Medical Experiments. It publishes ATLA. It also promotes the replacement of animals in laboratories through better science. Visit FRAME website
PiLAS is Perspectives in Laboratory Animal Science. It improves the quality of discussion about animal experimentation by giving scientists in all fields a place to exchange views. Read PiLAS