NS Magazine 2010

NEW SCIENCENEW SCIENCELord Dowding Fund for Humane Research

ADVANCED TECHNIQUES: News from the world of research without animals

Progress in cancer study � Implementation of alternativesInterview: Thomas Hartung � Cataract lens replacement � Animal testing directive

ISSN: 2042-9576

New Science l 2010 Lord Dowding Fund for Humane Research2

EDITORIAL

The Lord Dowding Fund

for Humane ResearchThe mission of the Lord Dowding Fund isto support, sponsor and fund advancedmethods of scientific and medicalresearch, which replace the use ofanimals; to fund non-animal medicalresearch (fundamental and applied) inorder to promote the adoption of non-animal research methodology; to fund,promote and assist learning and trainingfor the purpose of replacing animals ineducation and training.

Founded in 1973, the name of the Fundhonours Britain’s Air Chief Marshal theLord Dowding who, during his lifetime,was a President of our foundingorganisation, the National Anti-VivisectionSociety.

The Fund awards grants totalling anaverage of £300,000 a year to researchersworking in a wide range of fields.

Our projects have included cancer (breast,lung, metatastic); brain tumour, braindamage; brain toxicity; cataracts;ultrasound healing for cartilage; pain;vision; culture of tumours; infectionsincluding MODS and MRSA; computer-aided drug design; biotechnology;computer-assisted science learning;microsurgery techniques; toxicity testing ofdental fillings; safety testing; Parkinson’s;schizophrenia; cot death and kidneydialysis amongst others.

The Lord Dowding Fund is a department of the National Anti-Vivisection Society and Animal Defenders International.

New ScienceISSN: 2042-9576published by The Lord Dowding Fund for Humane Research

LDF UK: Millbank Tower, Millbank, LONDON, SW1P 4QP, UKTel: +44 (0)20 7630 3340Fax: +44 (0)20 7828 2179e-mail: [email protected] web: www.ldf.org.uk

LDF contact for USA:Animal Defenders International, 953 Mission Street, Suite 200,SAN FRANCISCO, CA 94103, USATel: +1 415 543 2344Toll free: 800 978 ADII (2344) (inside US)Fax: +1 415 543 2343e-mail: [email protected]: www.ad-international.org

South America: Apartado Postal 359888 BOGOTÁ, Colombiae-mail: [email protected]: www.ad-international.org

Editors: Creamer/PhillipsDesign: Creamer/Phillips/ElsonContributors: Jan Creamer; Tim Phillips; HelderConstantino; Christina Dodkin; Jessamy Korotoga;Amanda Gent.Cover image: LDF

©2010 LDF. All rights reserved. No part of this publicationmay be reproduced for commercial purposes by any meanswhatsoever without the written permission of LDF.

Welcome to the latest edition of New Science, which brings you the latest news from

the world of research without animals, as well as the project we support through the

Lord Dowding Fund.

People often ask “what’s THE alternative to animal experiments?”, as if research is

based on a single methodology. In reality, animal research is a backwater of the

world of scientific and medical research; the majority of research is conducted

without animals, using modern and superior technological and scientific methods.

Whilst animal experimentation is demonstrably limited, the options for non-animal

replacements are almost limitless.

Animal experimenters like to perpetuate the myth that there needs to be just one

alternative, with statements like “you can’t measure blood pressure in a computer”,

when they know full well that it is the use of a range of techniques in combination

which provides the replacement to animal use.

The regulators and politicians are comfortable with this simplistic one-model-

delivers-all notion. It fits easily into bureaucratic check-lists. As a result, the

implementation of replacements is restricted and slow. The current revision of the

European Directive on animal experiments has failed to properly address the issue of

replacements, notably in fundamental (basic) research.

This ignores the reality of research – that if we are to get the best scientific

endeavours, we need a raft of different techniques and we need to constantly develop

new technologies – more accurate and more relevant to people. Animal research is

the opposite of this; stagnant and scientifically unreliable. Tissue, organ and cell

cultures, sophisticated imaging techniques, computer analysis, simulations and

modelling such as QASARs, micro-dosing and accelerator mass spectrometry, as

well as toxicogenomics and others are all moving forward, leaving animal research

behind. It’s just that the regulators have not caught up, and in their anxiety about

new methods, they allow the animal experimenters to keep them in the dark ages.

The notion that experiments in a different species can deliver the information

required for the human situation is a fallacy. THE single alternative is a myth too.

We must embrace a multi-disciplinary approach that delivers the right answers, and

ensure that this is recognised and implemented by regulators.

That’s our approach with New Science: To showcase the best of new scientific

method and technology and to put this in the context of regulation and replacing

animal experiments.

Should people really accept substandard science and animal suffering merely because

regulators find implementation of the replacements too complex? We believe people,

animals, and science deserve better.

Jan Creamer

Chief Executive

Help Finance an EvolutionWill you leave a lasting legacy of compassion,

and help scientific advancement?

LDF finances scientific and medical research without the use ofanimals. The projects featured in this publication have only beenpossible thanks to the generous donations and bequests of our

supporters. Please consider a donation or legacy in your WILL to helpour work. Your legacy could advance medical research and help toend unnecessary animal suffering. Help finance an evolution and

leave a lasting legacy with a bequest or donation today, to the Lord Dowding Fund for Humane Research© Lord Dowding Fund

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Chief Executive, Jan Creamer, being interviewed at Aston Universityfor The OneShow.

New Science l 2010Lord Dowding Fund for Humane Research 3

NEWS

In the last issue of New Science we highlighted the innovative research by Dr. VehidSalih and Dr. Jamie Harle into a human, in vitro model of cartilage for testing thehealing properties of ultrasound.

Currently, Drs Salih and Herle are working on modifications to the ultrasound exposurerig to expand the range of acoustic outputs available. The best scaffold compositionwill be used in the ‘exposure’ work for the next stage of the project. This will enableexposure of pulsed ultrasound signals, rather than the continuous wave form used inthe work so far.

During the next phase of the work, researchers aim to investigate the biologicalresponses of hMSC (human mesenchymal stem cells) for both diagnostic andtherapeutic ultrasound exposure parameters. The exposure rig will be modified furtherallowing ultrasound signals to be amplified so that varying intensities of ultrasound canbe delivered to the hMSC-alginate constructs.

The model will then be used to look for ultrasound-induced effects which are intensity-,exposure time-, or pulsing regime-dependent.

As the use of donor grafts to solve orthopaedic problems is not ideal, producing naturaltissues artificially, with the use of such a scaffold to guide regeneration, is a goodalternative. The regeneration of bone tissue can be greatly improved by regulartreatment with therapeutic ultrasound. Therefore, the current research has greatpotential to develop these methods to greatly benefit patients with cartilage injuries.

The presentation was well received at the European Society for Biomaterials’ AnnualCongress in September 2009, promoting the use of HA(hydroxyapatite)/alginate as ananimal-free scaffold for an in vitro model of cartilage. The European Society forBiomaterials publishes a monthly journal and holds annual conferences to encourageprogress in the field of biomaterials.

NEWS IN BRIEFCosmetic industrymatches Commission infunding alternativesIn July 2009, the European Commissioncalled for proposals on the developmentof a strategy for alternatives to animaluse in repeat dose systemic toxicitytesting. The European cosmeticsindustry decided to match the researchfunds made available by theCommission, bringing the total availablefunds up to €50million.

The Commission will select up to sevenprojects to receive funding in areasincluding computational modelling,organ-simulating cellular devices andfunctional differentiation of human-basedtarget cells in vitro.http://www.nc3rs.org.uk/news.asp?id=1127

AltTox – promotingalternative methodsonlineAltTox (alttox.org) is a website whichaims to advance non-animal methods oftoxicity testing. The rationale for the siteis to provide better protection for humanhealth, animals and the environment. Italso aims to reduce the number ofanimals being used in toxicity testing andthe suffering that they undergo.

AltTox is composed of three differentparts:

Online forums – For posting news,information and for encouragingfeedback. The forums are split into fourcategories: ‘Toxicity Endpoints & Tests’,‘Emerging Technologies’, ‘Programs &policies’ and ‘Overarching challenges &opportunities’. Each forum’s moderator isan internationally recognised expert onthe subject.

Information on toxicity testing – TheTTRC (Toxicity Testing Resource Centre)is a comprehensive source of informationon non-animal methods of toxicitytesting. The topics are cross-referencedto the forums.

Invited commentaries – These areopinion pieces, written by experts, toaccelerate progress in the developmentof non-animal methods.

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Dr. Vehid Salih (left) and Dr. Jamie Harle and their ultrasound exposure apparatus (inset).

LDF Project Report: Cartilageresearch showcased atinternational conference

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As reported in the last issue of NewScience, LDF is funding exciting workcarried out by Professor Mike Coleman(pictured above) at Aston University. Thisresearch studies the reactions of braincells to toxicity, using a human-basedmodel to study neural function.

Most other cellular models use rapidlydividing cells, which does not occur inhuman brains where the cells are post-mitotic (they have stopped growing.)Professor Coleman’s model contains themost complete basic unit of the brain,the neurone/astrocyte combination. Thisis an improvement on many othermodels, which just look at neurones.

Ultimately, it should provide a better,more logical basis for re-creating theconditions of the neuronal/astrocytepartnership development. The modelmay also be used to understandneurodegeneration in the hope that thiswill lead to easier repair. As ProfessorColeman states “we hope that ourtechnique will provide scientists with anew and highly relevant humanexperimental model to help usunderstand the brain better and developnew drugs and treatments to tackleneurodegenerative disease.” This workrecently received extensive mediacoverage including BBC and Sky News.


NEWS IN BRIEFNew ECVAM advisory structure The European Centre for the Validationof Alternative Methods’ (ECVAM)Scientific Advisory Committee (ESAC) isto be renewed. ESAC, which is knownfor publishing opinions on the validity ofalternative test methods, will now gainsupport from the ECVAM Expert Pool(EPP), which is soon to be formed. AnECVAM Stakeholder Forum (ESTAF) willalso be created in order to maintain anactive dialogue with variousstakeholders, including those withspecific vested interests, includingvoicing societal concerns.

The new ESAC, comprising of senior,generalist scientists, will continue toissue opinions on ECVAM evaluatedalternative test methods. The EPP willconsist of a pool of highly specialisedscientists who will cover a range of areasnecessary for the validation of alternativetests, which would serve a wide range ofapplications.

ESAC working groups, dedicated to peerreview submissions received orcoordinated by ECVAM, will be chairedby ESAC full members with the groupcontaining mainly scientists from theEPP register. These working groups willprepare draft opinions for final adoptionthrough ESAC.

Coronary stent fails to live up to its promiseA new drug releasing coronary stent(tubular support placed inside a bloodvessel) was investigated by scientists ina ‘first-in-human’ study. ‘MAHOROBA®’was developed to treat coronary arterydisease.

The study used 47 patients sufferingstable and unstable angina and othercardiac problems. At six months, twopatients had suffered heart attacks and11 required repeated revascularisationbecause of ischemia (blood restriction)

One of the features of the tested stentled to “positive results” in pigs, but thisstudy “failed to establish theeffectiveness of the MAHOROBA®stent”. A report on the study concludesthat the “translation of results fromanimals to humans remains unclear” http://www.nature.com/nrcardio/journal/v6/n8/pdf/nrcardio.2009.117.pdf

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A neural network with astrocytes (shown in red).

LDF Project Report:Neurotoxicity researchgoes from strength to strength

NEWS


It has been stated that the “developmentof therapeutical [sic] monoclonalantibodies has resulted in an increasedneed for primates in reproductivetoxicology since the immune system inmacaques is much more similar tohumans compared with rodents.”1

However, as seen in previous disastroushuman drug trials such as TGN1412, theimmune systems of non-human primatesare not sufficiently similar to those ofhumans to detect human immunereactions. The importance of testinghuman immune reactions to manymodern biopharmaceuticals hasemerged due to the very high degree ofspecies specificity of drugs.

The lymphatic system is involved inhuman immunity2. Lymphatic fluidtransports bacteria and other foreignagents along the lymph vessels to thelymph nodes. Lymphatic fluid ispresented to antigens held in the nodesas it is filtered through them. As fewervessels leave the node than enter, flowthrough the nodes is slow, allowing timefor foreign agents to be destroyed by theantigens3. Lymphocytes, found in thelymph node, are antigens which areprimarily responsible for specificimmunological responses4.

As animals possess an often significantlydifferent immune system, it is insufficientto carry out tests with any animalspecies to assess the efficacy and safety

of drugs in humans. The creation of afully human Artificial Lymph Node (ALN)circumvents “the limitations associatedwith the use of animal models”5.

The ALN model uses a bioreactor whichis able to control the environment of thetissues being grown. The model is alsoreproducible which is important for itspossible role in drug development andscreening. The ALN mimics thephysiology of the lymph node in thehuman body by integrating mobile andimmobile cell phases to ensure the cellsinteract effectively6.

A perfusion of cellsand media flow froman outer culture spaceinto a central culturespace, which is a fixedmatrix containingimmobile cells. Thisallows cells flowingthrough the system tobe presented to thefixed cells. Thisemulates the actionswhich occur in thelymph node when thelymphatic fluid flowsinto the node, passingby the lymphocyteswhich carry out animmune response. Asthis system creates anenvironment which is

similar to that of the lymph node, it willbe useful for looking at the effects ofdrugs and cell therapies7.

Dr. Uwe Marx, one of the key scientistsin the development of the ALN model,purports that in order to develop andadopt such models across the EU, a“human in vitro organ” program shouldbe designed, specifically “for predictivereliable substance testing” incorporatingdisciplines such as tissue engineering,bioreactors, and computational models8.

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Above: Wild, freemacaque.

1. http://ec.europa.eu/health/ph_risk/committees/04_scher/docs/scher_o_110.pdf 2. http://uhaweb.hartford.edu/BUGL/immune.htm#top 3. Marieb, E.N (1992) Human Anatomy and Physiology 2nd Edition,The Benjamin/Cummings Publishing Company, Inc. California 4. Miller, J.F.A.P, Lymphocytes, In: Dulbecco, R (Ed) (1991) Encyclopedia of Human Biology, Vol. V, Go-Me, Academic Press, Inc USA 5.http://www.probiogen.net/english/news/view_newsfile.php?id=134 6. Pörtner, R and Giese, C, An Overview on Bioreactor Design, Prototyping and Process Control for Reproducible Three-Dimensional TissueCulture. In Marx, U and Sandig, S (2007) Drug Testing In Vitro: Breakthroughs and Trends in Cell Culture Technology, WILEY-VCH, Germany 7. Hitchcock, T and Niklason, L (2008) Lymphatic TissueEngineering Progress and Prospects, Annals of the New York Academy of Science Vol. 1131, pp: 44-49 8. Pers. Comm. Dr Uwe Marx 14/04/08

Artificial Lymph Nodemedia exchange cycled cell suspension

immobilecell-network

porous membrane

gas, media,substrates

matrix

mobile cells

HFM

PCS

CCS

Concept of mobile and immobile cell phases interacting in the artificial lymphnode bioreactor. The suspended lymphocytes pass the CCS and come intoclose contact with the immobile dendritic cell network in the embeddingmatrix of the CCS. (HFM = hollow-fibre membrane. PCS = peripheralculture space. CCS = central culture space.)

Artificial Lymph Node:Testing humanspecific immunereactions

Artificial Lymph Node:Testing humanspecific immunereactions

LDF PROJECT: ARTIFICIAL LYMPH NODE


EXPERT OPINION

New Science (NS): What does yournew role at CAAT involve? How doesthis differ from your previous role atECVAM (European Centre for theValidation of alternative Methods)?

I would characterise it as two sides ofthe same coin: both institutions havebeen promoting alternative methods formany years. ECVAM is giving scientificpolicy support, especially to theEuropean Commission, and isresponsible for the formal validation ofmethods; CAAT has, for 28 years, beena key information hub sponsored byprivate foundations and industry. CAAT isnot working primarily on validation. Aspart of Johns Hopkins, one of the leadinguniversities worldwide, it is primarilybound to sound science and morebroadly to humane science approachesand the paradigm shift in toxicology. Iaim to further develop CAAT towardbecoming a think tank of toxicology. Lastbut not least, education of students is akey role of an academic institution andwe try to promote humane science invarious curricula.

NS: What motivates you to beinvolved in the development ofalternatives to animal testing?

I want to move the science of riskassessment forward and make it morehumane, so it is about safety of patientsand consumers AND animal welfare. Ibelieve that alternative methods are theforerunner for the revolution of riskassessment sciences ahead of us.Alternatives are the pilots for a newtoxicology and for quality assurance inthe life sciences in general. There is noother field, where the same effort isspent to ascertain the predictive value ofmodels and to quality control them.That’s where science has to move – weare publishing far too much, which is notreproducible, is irrelevant or simplyartefact.

NS: What barriers, do you think,prevent researchers in academia andworking in pharmaceutical companiesfrom using alternatives as opposed toanimals?

There are a few barriers: traditions,regulatory requirements, lack ofinformation on opportunities. However,on a positive note, there is also pressurefor change such as the biotech andinformatics revolution, which providednew tools, companies commercialisingthese, legislative pressures such as theEuropean animal welfare legislation,REACH and the 7th amendment of theCosmetics Directive [total ban on animaltesting regardless of availablealternatives]. Furthermore, needs forhigher testing throughput and increasingawareness of the shortcomings ofcurrent tests help a lot.

NS: What are the advantages of usingnon-animal methods in toxicology?

Simply, to overcome the limitations ofanimal testing. These are not only ethicalones: animal tests are costly, have lowthroughput and do often give wrongresults. Some were developed 40-80years ago, especially in regulatorytoxicology. In the meantime a body ofknowledge has been built onmechanisms, molecular targets andmodes of action. These must be fullyexploited before targeted, hypothesis-driven testing in animals starts, if stillnecessary.

NS: Do you think there will be a timewhen animal testing will not be a legalrequirement in the regulatory testingof drugs?

Yes – we already see that we areharming the drug development processwith wrong decisions. Killing valuablesubstances under development also killspeople. Furthermore, more than half ofthe new drugs are human proteins orantibodies to human proteins making

classical approaches useless. If we canshow that we can handle these properlywithout animal testing, why shouldn’t wedo so for small molecules?

NS: Do you believe that the use ofprimates in experiments can bereplaced and, if yes, how?

In the end by moving to human testing,therefore bypassing non-humanprimates, e.g. microdosing. This certainlyrequires another level of certainty in thenon-animal methods and non-primatemodels. The trend to use humanproteins is certainly prompting a lot ofprimate use. However, the tragedy of theTegenero antibody [TGN1412], whichhad no effect at 500 times higher dosesin primates and almost killed instantly thefirst six volunteers treated, has shownthat also this testing has questionableresults.

NS: Do you regard the new Directiveon animal testing, as drafted by theEuropean Commission, as a good textto regulate animal testing andpromote alternatives?

Europe has, since 1986, had the mostprogressive legislation for animal welfareworldwide. The new draft is moving thiseven further – exactly how far will bedetermined by the political decisionprocess. It will certainly raise standardsfurther but also leave room forimprovement for the next revision…

NS: Would you agree with thestatement that the use of animals inexperiments is a scientific dogma thatmust be challenged?

I do not consider it a dogma – somethingbelieved to be true without question. Iconsider it a tool with increasingly knownlimitations. The more we make theselimitations obvious and develop othertools, the less useful, and less used, theanimal tools will become.

For a full version of this interview, pleasevisit www.ldf.org.uk

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Dr. Thomas Hartung, Director of the JohnsHopkins Centre for Alternatives to AnimalTesting and former Head of ECVAM, inconversation with New ScienceDr. Thomas Hartung is a leader in the field of toxicology who was, in 2009, appointedDirector of the Johns Hopkins Centre for Alternatives to Animal Testing (CAAT). As anadvocate for alternatives to animal testing he has assisted in the development of severaltest strategies, revolutionising the way in which chemical safety assessments are carriedout in Europe. In this exclusive interview, Dr. Hartung reveals his motivations and hopesfor the future of advanced methods to replace animal testing.

strategies can help toovercome some ofthese obstacles.Networking betweenstakeholders andinvolving regulators inthe validation processand research programshave also beensuggested as ways ofalleviating some ofthese problems3.

Another ECVAMworkshop pointed out the difficulties ofusing traditional test methods, such as invivo, as reference against which tocompare new alternatives. It is pointedout that assumptions about the toxicity oftest compounds are based on these‘traditional methods’ which may or maynot accurately reflect human data. Thereport of the workshoppoints out that “themajority of thesetraditional teststhemselves were nevervalidated” and that,although traditional testmethods are broadlyaccepted, they do notnecessarily qualify asreference testmethods4.

Some useful methodsare being used by the EuropeanCommission to monitor/track theprogress of alternative tests, such as theTracking System for Alternative TestMethods Review (TSAR). This systemtracks methods from the point when theyare purely scientific protocols submittedfor pre-validation, to their active use in aregulatory context5.

Many scientists areresearching methodswhich are able, or havethe potential, toprovide alternatives.However, the reality isthat theirimplementationdepends on more thanjust development.

Many scientists, such as those funded bythe LDF, are working to develop methodswhich could and should be used in placeof tests on animals. However, the actualregulatory acceptance, implementationand dissemination of these methods is along and sometimes difficult process.

The validation process

The European Centre for the Validationof Alternative Methods (ECVAM) wasestablished in 1991 as part of theEuropean Commission’s Joint ResearchUnit. The duties of ECVAM include: thevalidation of alternatives; being a focalpoint for the exchange of information ondevelopment of alternatives; to promotedialogue between legislators, industry,scientists, consumer organisations andanimal welfare groups on the issue ofalternative methods1.

The ECVAM validation process involvesestablishing the reliability and relevanceof an alternative procedure for a specificpurpose in five stages2:

1. Test development – The test must bedescribed and its scientific purposedefined. A protocol must be producedwhich is comprehensive enough thatthe test may be carried out by anotherlaboratory (interlaboratorytransferability).

2. Prevalidation – This stage optimisesand standardises the protocol. It isimportant for ensuring that methodsentering a formal validation study havea greater likelihood of fulfillingexpectations of scientists andregulatory bodies.

3. Validation – An interlaboratory blindtrial is carried out at this stage, with‘preliminary’ and ‘definitive’ stageswhere coded chemicals are tested.Finally, data is analysed and theoutcome of the studies evaluated.

4. Independent assessment – Thepublished results of the validationstudies are considered by one or moreindependent assessment panels.These panels need to be independentof the validation study in question andrepresentative of the scientific,toxicological, industrial, regulatory and


animal welfare communities. Amongother aims, the panel should considerwhether the purpose and objectives ofthe validation study have been met.

5. Progression toward regulatoryacceptance – The validated methodsneed to receive widespreadinternational recognition. For examplethrough the Mutual Acceptance ofData Agreement, the OECD(Organisation for Economic Co-Operation and Development)agreement sets out that membercountries agree to accept data fromtests which are performed to OECDtest guidelines. These guidelines areused in Europe, Japan and NorthAmerica.

Validation and acceptance problems

Recent publications have focused on theneed to ensure that once methods havepassed the validation process they areaccepted and implemented by regulatoryauthorities. An ECVAM workshop on theoptimisation of the post-validationprocess highlighted some obstacles toregulatory acceptance of validatedalternative methods:

� Reluctance to share data by some

companies as it may harm theircompetitive position. Data sharingwould, however, avoid duplication.

� Insufficient communication, for

example, between scientists andregulators.

� The tendency of regulators to favour

refinement and reduction overreplacement, due to a familiarity withanimal procedures and the ease ofmodifying these instead of replacingmethods altogether with alternatives.

� Scientists lack knowledge about the

regulations and the opportunitieswhich could influence thoseregulations.

There are ways to overcome theseobstacles. By taking advantage of jointinitiatives such as the EuropeanPartnership for Alternative Approaches toAnimal Testing (EPAA) the mapping ofexisting research and development of

Guide to validation andimplementation of alternatives

VALIDATION AND IMPLEMENTATION

1. http://ecvam.jrc.it/page_pdf.cfm?voce=m&idvoce=1 Acessed 29/09/09 2. Balls, M et al (1995) Practical Aspects of the Validation of Toxicity Test Procedures: The Report and Recommendations of ECVAMWorkshop 5, IATLA 23, 129-147 3. Bottini, A. A et al (2008) Optimisation of the Post-validation Process: The Report and Recommendations of the ECVAM Workshop 67ª, ATLA, 36, 353 - 366 4. Hoffman,S et al (2008) Points of Reference in the Validation Process: The Report and Recommendations of ECVAM Workshop 66ª, ATLA 36, 343 - 352 5. http://tsar.jrc.ec.europa.eu/ Accessed 12/10/09

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Above and below:Non animal testingmethods in use forthe neurotoxicityresearch project atAston University, asdescribed on p4.

Below: AcceleratorMass Spectrometry(AMS) equipment inuse at GlasgowUniversity. AMSoffers preciseanalysis of samplesfrom volunteerswho have receivedmicro-doses of testcompounds.


the BBB a highly effective biologicalbarrier so these results indicate thatastrocytes improve the TJ formation.

Currently, the addition of pericytes tocreate a tri-culture has not improved tightjunction formation. However, this ispossibly due to the pericytes blocking theendothelial cells, preventing them fromproducing a monolayer. The team intendto repeat the experiment, with thepericytes at the bottom of the Transwell®so that they are not in direct contact withthe endothelial cells.

It is hoped the model will identify thepathways malignant cells take to crossthe BBB and, once fully developed, couldbe used to study metastases. Inaddition, because of the importance ofthe BBB in conditions of the CentralNervous System, such as stroke andbrain trauma, this model has thepotential to improve the treatment ofthese conditions. Excitingly the modelcould also be used as a tool by drugcompanies to discover if, and how,therapeutic agents pass through theBBB.

With 25% of cancers found in the bodyspreading to the brain and worseningpatient prognosis, it is vital to understandthe mechanisms by which cancerouscells metastasise (move) into the brain,thereby spreading the cancer. In vitroconstructions generally use animaltissues, such as rat and mice cells, tomodel the blood-brain barrier (BBB), thelayer which covers and protects thebrain. However, these animal models donot reflect the situation in humans.Those models which do use human cellsare often grown in media which includefoetal calf serum, a product which isethically unsound.

Exciting new progress has been madehowever, in the development of an all-human tissue model of the BBB atPortsmouth University, where ProfessorGeoffrey Pilkington and his team havebeen testing various permutations of cellcultures in the Transwell® model. Theresearchers are using astrocytes(supporting cells) from two differentareas of the brain; endothelial cells,which line the blood vessels, andpericytes. Pericytes are a less wellknown component of the BBBand are thought to regulateproliferation and differentiation ofendothelial cells. Many otherresearchers do not includepericytes in in vitro models dueto the difficulties involved byintroducing them into the BBBmodel. However, the team atPortsmouth has persevered withthis element as they believe theirinclusion is needed to produce atrue reflection of the in vivosituation.

The Transwell® model is set upby adding the human astrocytes

and endothelial cells to either side of amembrane filter. This membrane iscoated with extracellular matrices (ECM)and the whole model is grown in humanserum supplement.

The initial stages of the study wereperformed to analyse the components ofthe BBB model, to aid the developmentof the Transwell® model. In these firststages, different cancer cells were addedto various cultures of brain cells. Resultsfound lung cancer cells, which wereobserved over 48 hours, to movethrough an endothelial cell mono-culture,demonstrating the cancer’s highmetastatic potential.

Aspects of the cells were measured andobserved using state of the arttechniques such as ECIS™, which useselectrical currents to measure theresistance of cells and cell behaviour,treatment with immunoflorescence andtime lapse microscopy.

Recent preliminary results have revealedthat co-cultures of astrocytes andendothelial cells have shown betterformation of ‘tight junctions’ (TJs) thanendothelial cell mono-cultures. TJs make

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Progress in braintumour projectProgress in braintumour project

insert

Astrocytes

Endothelial cells

Endothelial cell growth medium

Astrocyte growth medium

Membrane filter (white)ECM components (blue)

Professor Pilkington’s Blood-Brain Barrier

LDF PROJECT: BLOOD BRAIN BARRIER

Above: A slideshowing stainedendothelial cells,which line the bloodvessels.

Cataracts are cloudy patches in the lensinside the eye and are the main cause ofimpaired vision in the world, with anestimated 30% of over 65 year oldshaving cataracts in one or both eyes.There are various risk factors for thecondition, which include a family historyof cataracts, smoking, poor diet andexposure to sunlight1.

According to an article in the BritishMedical Journal, “cataract surgery ispossibly the world’s oldest surgicalprocedure”, being brought to Europefrom India by the armies of Alexanderthe Great2.

The only cure for cataracts is to removethe lens from the eye, which is done byeither surgically removing the nucleus ofthe lens as a whole or by ultrasonicallyfragmenting the nucleus, which is thensucked out of the eye. The latterprocedure requires a smaller incision,causes less inflammation post-operatively and rehabilitation takes 1-3weeks, rather than 2-4 months for theformer process.

Once the affected lens has beenremoved, it is replaced with an intraocular lens (IOL). However, some lens’epithelial cells still remain after thesurgery. In approximately 25% ofpatients this leads to the development ofPosterior Capsule Opacification (PCO), acondition where the epithelial cellsspread across the visual axis of the eye.The design of IOLs can help preventPCO, but current research into IOLdesigns is largely conducted on rabbits.

LDF sponsored research by Dr. MichaelWormstone at the University of EastAnglia is using ‘eye bags’, from humandonors, to carry out such tests. Dr.Wormstone recently established theoptimum level of human serum in whichto maximize cell growth on the lens. Thiswas done by counting cell growth in

tissue culture at varying concentrationsof serum.

Once this was established, the first IOLwas implanted into an eye capsule. Aneye bag without an IOL was used as acontrol. The observations, to monitor anysign of PCO indicated by cell re-growth,were made using phase-microscopy. TheIOL did slow the development of PCO,but did not prevent it completely; cellsgrew in sufficient number to cover theposterior of the capsule. Wrinkling of the

capsule’s posterior also occurred, butless severely. In the human patient thiswould have resulted in impairment of thepatient’s sight.

The initial IOL had round edges and it isknown that IOL designs with a squareedge are better at preventing PCO.Therefore, the next lens to be tested willbe square-edged IOL which, if there isseen to be further retardation of cellgrowth, will provide further evidence forthe use of the current model.


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Cataract lensreplacement tested in fully human system

LDF PROJECT: CATARACTS

1. http://www.cks.nhs.uk/patient_information_leaflet/cataracts_age_related 2. Spalton, D.S & Koch, D (2000) “The Constant evolution of cataract surgery”, BMJ ,vol.321. pp. 1304

Below: A piece oflens cultured inhuman serum. Cellgrowth can be seenin purple.

Lord Dowding Fund for Humane Research

LDF PROJECT: EDUCATION

The latest figures from the 25 EUMember States, reporting animal use in2005 (2004 for France), state that198,994 animals were used for“education and training”. This isstaggering in its enormity and also thediversity of species used in this area.The report gives scant information on theexact use of these animals, whichinclude 86,597 mice, 50,048 rats, 3,856rabbits, 5,854 pigs, 1,391 cattle, 15,666amphibians and 23,796 fish.

The figure fell since the precedingreporting period, 2002 (2001 for France)which reported that 341,967 animalswere used. The latest report states thatthe “decrease of animals used foreducation and training can be attributedto both an uptake of alternativetechniques and the re-use of animals.”However, we believe that many moreanimals are being used in dissections orother procedures where statistics are notpublished.

LDF is funding an exciting new project toclarify exactly what these animals arebeing used for, to gain valuableinformation regarding their use and alsoto target the implementation ofalternatives to certain subject orgeographical ‘hot-spots’.

This is the first LDF funded survey, and itwill be on a massive scale! The aim ofthe survey is to find out exactly whereanimals are being used in highereducation, for which subject and inwhat numbers. In addition, theavailability and

implementation of alternatives willbe investigated. The results of theresearch, which is being overseenby Professor David Dewhurst at theUniversity of Edinburgh, will be aunique tool for targetedreplacement of animals ineducation. In addition, it willdemonstrate the feasibility ofgathering quantitative data in orderto assess possibilities for theimplementation of advanced techniquesin other areas of research where animalsare currently used.

Questionnaires will be sent touniversities and professional bodies inthe target member states, to gatherinformation on the use of alternatives toanimals and specific data regardinganimals, such as where they are usedand why. The researcher conducting thesurvey will follow up the questionnaireswith phone calls and emails to maximisethe number of responses, thereby givingLDF a more detailed picture of thesituation.

At the end of the survey it is hoped thata comprehensive picture of the use ofanimals across the EU will have beengained, along with information on howand where LDF can push alternatives inorder to reduce the number of animalsused from 200,000 to ZERO.

We will report on the progress ofthe survey, and its outcomes infuture editions of NewScience and in our sister

magazine Animal Defender.

EU survey of animal usein education

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New Science l 201010

LDF and Professor David Dewhurst – 24 years of Progress

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Left: Dewhurstcomputer, 1988.

Below: Earlycomputersimulation, 1993.

Below: Prof. Dewhurstreceiving the Queen’sAnniversary Prize forHigher and FurtherEducation, 2005.

Right: Workshop in India, 2006.

Below: Prof. Dewhurst receives theDoerenkamp-Zbinden FoundationInternational Prize for Animal FreeResearch, 2006

Right: The physiology andpharmacology simulation,having been subjected tothe RECAL process, 2009.


LDF PROJECT: EDUCATION

1986 LDF’s funding of Dr. Dewhurst began in order to strengthen the NAVS Violence Free

Science campaign. Nerve physiology simulation for BBC microcomputer released

1987-9 Muscle physiology, Frog Heart, Neuromuscular Pharmacology and ‘Rat stack’ (ratdissection) simulations released

1989 New projects to develop computer-based alternatives in pharmacology and rat dissection(interactive video-disk) commenced. Conversion of BBC microcomputer versions to IBM(DOS) versions

1991 Replacement of all frog, rat, rabbit and guinea pig practicals with computer simulations atLiverpool and Leeds Polytechnics. Several new simulations released including Guinea PigIleum, Cat Nictitating Membrane, Frog Skin, Intestinal Absorption in Rat

1993 Evaluations of educational effectiveness of the simulations carried out in small number ofUK universities – all supported the view that students’ learning using simulations was atleast comparable to that in animal laboratory classes

1996 One of the simulations, Rat Blood Pressure wins the 1996 Multimedia Festival held at the2nd World Congress on Alternatives and Animal use in the Life Sciences, Utrecht, TheNetherlands

1997 Megan Quentin-Baxter, who developed the interactive video-disk of a rat dissection, wasawarded her PhD which was largely based around this work

1998 Work commenced on rewriting simulations for the MS Windows environment alongside thedevelopment of new simulations

1999 New project supported by LDF to continue redevelopment of simulations for MS Windowsbegan at the University of Edinburgh

2001 The European Resource Centre for Alternatives in Higher Education (EURCA) is set up byDavid Dewhurst (University of Edinburgh) and Jan van der Valk (University of Utrecht) withthe help of LDF and the Doerenkamp Zbinden Foundation

2003 The funding of RECAL began, which should eliminate the need for rewriting computerprograms to keep pace with technological advances and enable teachers to assemblepersonalised teaching and learning materials for replacing animals in higher education

2004 A lecture tour of Brazilian Universities was carried out by Professor Dewhurst to promotecomputer-based teaching packages to replace animals

2005 Professor Dewhurst’s team awarded the Queen’s Anniversary Prize for Higher and FurtherEducation, for their work in using e-learning to support medical and veterinary education inEdinburgh

2006 Workshops held in China and India to promote computer alternatives to higher educationprofessionals. A workshop was also held for teachers of physiology and pharmacology atBalkan Universities. Professor Dewhurst awarded the 2006 Doerenkamp-ZbindenFoundation International Prize for Animal Free Research at a ceremony in Linz

2007 With seventeen computer programmes having been successfully taken apart andsubjected to the RECAL process, further funding was granted for the project’s continuation

2010 LDF continues to fund Professor Dewhurst’s projects at the University of Edinburgh

LDF and Professor David Dewhurst – 24 years of Progress©

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Below: Prof. Dewhurst atBalkan University, 2006.

New Science l 201012

The new Directive’s regulations willrepresent a significant step forwardfor animal welfare compared to thoseof 86/609/EEC, defined nearly aquarter of a century ago. However,whether the Directive will have apositive impact on those who arepromoting, developing and using non-animal research methods remains tobe seen.

The European Commission’s proposalon the use of animal testing, issued inNovember 2008, has entered the finalstage of the EU legislative process. TheCouncil of Ministers, chaired by theSwedish Presidency, and the Rapporteurof the European Parliament ElizabethJeggle MEP (Germany, EPP) reached acompromise in December 2009.However, the political groups of theEuropean Parliament have yet toapprove the Council’s amended text. Infact, the Greens/EFA Group hasannounced that it will reject the currenttext, with one of the major issues atstake being the weakening of therequirement to use alternative methods.Yet, their promotion was originally one ofthe main aims of the new Directive.

In Recital 11 of the version presented bythe European Commission in 2008, theDirective states that “the principles ofreplacement, reduction and refinementshould be implemented through a strict

hierarchy of the requirement to usealternative methods.” Thus, the textstarts with a very positive signal thatreplacement is the highest goal of theDirective.

The ‘3Rs’ principle is implemented inArticle 4: “where a method of testing notinvolving the use of animals exists andmay be used in place of a procedure,Member States shall ensure that thealternative method is used.”

In paragraph 2, the principle of reductionis laid out, followed by refinement inparagraph 3; both apply as soon asanimals are used, including in breeding,accommodation and care. However, thereplacement principle in the newDirective is not new, it was included inArticle 7 of the 86/609/EEC Directive, inequally strong terms: “an experimentshall not be performed if anotherscientifically satisfactory method ofobtaining the result sought, not entailingthe use of an animal, is reasonably andpracticably available.”

The real innovation of the Commissionlies elsewhere. In order to accelerate thedevelopment and validation ofalternatives, the Commission createdNational Reference Laboratories (NRLs)in Article 46: “each Member State shall(…) designate a national referencelaboratory for the validation of alternativemethods replacing, reducing and refiningthe use of animals.” The article detailsspecific requirements and objectives toNRLs. It states they must be staffed withpersonnel qualified in alternatives andhave appropriate infrastructures and

equipments. In addition their function isto participate, in coordination with theCommission, to: validate alternativemethods; provide scientific assistanceand information to authorities; andprovide training in alternative methods.

LDF welcomed this proactive approachwhich could multiply the number ofvalidated alternatives and contribute toalleviate the bottleneck at the EuropeanCentre for the Validation of Alternatives(ECVAM). ECVAM was created by theCommission to support EU objectivescontained in Directive 86/609/EEC. Dueto lack of funding ECVAM has onlyvalidated 27 methods since its creationin 1991, since it could only review asmall proportion of the alternativemethods submitted. This bottleneckconsiderably slows down ethical andscientific progress in Europe.

Caroline Lucas MEP (UK, Green Party)criticised it in October 2008 in aparliamentary question which said that“the Commission has repeatedly statedthat it would not allow validation tobecome a ‘bottleneck’ in the process ofbringing new non-animal test methodsinto use, and yet it appears that this isexactly what has happened.”

NRLs could be seen as theCommission’s response to criticism: astep to diversify centres for validation. Itwas, however, an incomplete responseas the first casualty of NRLs wasECVAM itself. ECVAM has a fragile legalexistence: it is based on acommunication from the EuropeanCommission. The Directive was anexcellent opportunity to strengthen it, but

the Commission failed to mentionit in its draft. Moreover, the fundingenvisaged for NRLs in theCommission’s Impact Assessmentof the Directive was extremely low:approximately €100,000 per MemberState. How can such a tiny budgetovercome ECVAM’s shortcomingswhen the latter, already verybacklogged, receives about €2millionper year? When the draft Directive went

EU DIRECTIVE ON ANIMAL EXPERIMENTS

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Opposite, from mainpicture, clockwise:Jan Creameraddresses theProgress WithoutPain conference atthe EuropeanParliament.Alongside (from left)are CommissionerDimas, Neil Parish MEP,and LDF grantholder ProfessorPaul Furlong.

The EuropeanCommission.

Amongst ADI’ssupporters in theEuropean Parliamentwere Jens HolmMEP, Caroline LucasMEP, Mojca DrcarMurko MEP.

ADI CampaignsDirector Tim Phillipsand Head ofParliamentary AffairsHelder Constantinoleave Parliamentafter giving oralevidence to theHouse of LordsCommittee reviewingthe new EU rules forlab animals.

Below: Alternativeresearch methods inprogress at AstonUniversity; ADIproduced manyscientific and legalTechnical Briefingsin up to 12languages.

New dawn orbureaucratic impasse?New dawn orbureaucratic impasse?

Lord Dowding Fund for Humane Research

New Science l 2010Lord Dowding Fund for Humane Research

to the European Parliament foramendments, LDF successfully lobbiedto improve the Commission’s text.ECVAM was firmly anchored to the newDirective with a new Article 45a, aimedat coordinating NRLs, but also continuingto validate alternatives and research intotheir development etc. In that framework,NRLs would not be created at theexpense of ECVAM but would worktogether.

An amendment was voted to strengthenthe duty for Member States to contributefinancially to alternatives. Thismechanism was completed by twoamendments creating regular thematicreviews. This key proposal from AnimalDefenders International, on behalf ofLDF, aims at keeping replacement ofanimal testing on the political agenda,instead of waiting another 25 years fornew legislation.

A new Article 53a was created, whichgives the Commission the duty to“conduct a thematic review of the use ofanimals in procedures every two yearscommencing two years after the entryinto force of this Directive. The reviewshall examine the impact ofdevelopments in technological, scientificand animal welfare knowledge, and settargets for the implementation ofvalidated replacement methods.” Theconcept of targets to replace animaltesting was therefore introduced inlegislation for the first time, which was agroundbreaking achievement. A verysimilar amendment to Article 8 onprimates was passed to have reviewsand targets specific to these species.

Unfortunately, this progress is now injeopardy in the Council of Ministers, whocan amend the Directive, following thevote in first reading in the EuropeanParliament (which took place in May2009). The Swedish Presidency cameunder strong pressure from key MemberStates, themselves lobbied by the pro-

vivisection industry, to undermine therequirements to use alternatives.

Firstly, the very cornerstone of the dutyto use available alternatives wasdamaged. A new Article 13 was added tointerpret the principle of replacement asthe duty to ensure “that a procedure isnot carried out if another method ortesting strategy for obtaining the resultsought, not entailing the use of a liveanimal, is recognised by Communitylegislation.” Originally, the principle wasto use any existing alternatives. Thisdrastic narrowing of the scope of theduty to use alternatives is a huge stepback, not only from the Commission’sdraft, but from the old Directive86/609/EEC; both provided a muchbroader obligation.

The consequence is that only the handfulof methods validated by ECVAMconcerning cosmetics and toxicity testingwill be strictly compulsory.

ECVAM disappeared and was replacedby a ‘Community Reference Laboratory’,responsible for “coordinating thevalidation of alternatives” and “exchangeinformation”. Although it can still validatemethods, it lost its remit with regards toconducting and commissioning researchinto new alternatives, as well asimplementing strategies to put the 3Rsinto action.

The NRLs also disappeared, and werereplaced by a general duty for MemberStates to contribute to the validation ofalternatives and “assist the Commissionin identifying and nominating suitablespecialised and qualified laboratories tocarry out those validation studies.” Thesetherefore have a very similar profile toNRLs, but again with fewercommitments. For instance, NRLs werecreated to validate but also providetraining and technical assistance: thiscrucial aspect has been removedfollowing the Council’s amendments. Thethematic reviews survived but are, in the

Council’s version, to belaunched whenever theCommission wishes,instead of biannually.

These amendmentscombined may put thedevelopment ofalternatives in Europe ina bureaucratic impasseand will allow someindustries anduniversities to continueusing animals whereadvanced methods areavailable, simplybecause there will beinsufficient means tovalidate them.

This situation is due tothe lack of vision fromsome key MemberStates, who areunwilling to fund theeffort into alternatives,despite potentially hugebenefits for science andresearch in the future.

The Directive isdescribed in Recital 8as “an important steptowards achieving thefinal goal of fullreplacement ofprocedures on liveanimals”. However, ifthe situation is notimproved in the laststage of thenegotiations, these willremain empty words.There is still room fornegotiations and therewill be opportunities totable amendments inthe EuropeanParliament. LDF will notfail to continue to lobby,for the sake of scienceand animals.

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According to the latest EU statistics,over 12 million animals were used inexperiments in one year1. It is vital, forthe clarity of research and publicaccountability, for every EU citizen tobe able to see, preferably at a glance,exactly what level of pain, sufferingand stress these animals endure.Disappointingly, the proposedrevision of the EU Directive on animalexperiments and the amendments byMEPs lack any solid, scientificallybased categorisation of the pain andsuffering that animals experienceeach year in the EU Member States.

On the basis of the EuropeanParliament’s amendments, the EuropeanCommission convened an expert workinggroup to address this issue (ADI did notparticipate). The group met for two daysin July 2009 and came to a consensuson the classification of variousprocedures and also a minimum level ofsuffering which needed to becategorised, but did not stipulate amaximum suffering level for animals. Theprinciple of a maximum level of sufferingis used in New Zealand. It states in theAnimal Use Statistics that “it may bedecided that the impact of a procedure

on the animal is so great that itshould not proceed, no matter whatthe potential benefit is.”2

Whilst we oppose all animalexperimentation as unnecessary,unreliable and unscientific, untilanimal experiments are a thing ofthe past the reality of life inlaboratories should be clear for all tosee. A transparent and trulyreflective system, which realisticallyportrays the suffering of laboratoryanimals, is one step towardsensuring this.

ADI, unconvinced of the robustnessof the classification system whichwas formulated, wanted to providepolicy makers and other interestedparties with a comprehensivealternative which accurately reflectsthe situation for millions oflaboratory animals. So we publishedthe ‘EU Severity ClassificationSystem’, an outline of which is givenhere.

The ADI classification system hastwo elements: severity classes andcomponents of suffering, which arederived from systems currently inuse in New Zealand, Switzerland,Canada and the UK.

The titles of the severity categorieshave been chosen to be reflective ofthe actual situation the animal isexperiencing without being emotive.It is vital that any system ofcategorisation is easily understoodand provides an accurate portrayalof suffering, even to the generalpublic.

The proposed categories of severityare: mild, consequential, significant,severe (transient), severe andprolonged or non-transient (which willnot be authorised). We believe thatthese are more reflective of thesituation than mild, moderate andsevere.

Classification is based on the durationand intensity of pain, suffering, distressand enduring harm. It also depends uponthe frequency and number ofinterventions, the shortfall in the animal’sbehavioural needs and whether or notanaesthesia and analgesia are used.The severity categories represent themaximum impact / maximum severitythat will be suffered by a single animalthroughout the procedure. Repeatedinterventions or those in combination canhave the effect of increasing the project’sseverity. Actual suffering rather thanintended suffering should be identified,reported and made public.

The components of suffering are theaspects of an animal’s life which must besatisfied in order to maximise itswellbeing; any diversion from the norm inthese areas can affect the animal, itswellbeing and the data gathered from it.

The components of suffering are:

� Mental state and dignity – animals

should not suffer fear, stress orundignified treatment. Mental sufferingshould be avoided

� Food and water – animals should be

free from thirst and hunger withaccess to fresh water and a dietenabling the maintenance of full health

� Environmental challenges – the

animal’s environment should notcause it discomfort

� Injury / Functional impairment /Disease – these should be kept to aminimum by ensuring rapid diagnosisand treatment. The degree of induceddisease should be kept to a minimum.All measures to minimise andameliorate pain and suffering shouldbe implemented

� Behaviour – the animal’s behaviour

should reflect its natural behaviouralrepertoire as closely as possible.

ADI publishes EU SeverityClassification System Briefing

© Louise Wallis / Animal Defenders International

Right: These ratsare sufferingexcruciating pain;their legs areswollen andinflamed withartificially-inducedarthritis. It is vitalthat the EuropeanSeverityClassificationSystem introducesan upper limit onlevels of pain whichcan be inflicted.

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1. http://ec.europa.eu/environment/chemicals/lab_animals/pdf/staff_work_doc_sec1455.pdf

2. http://www.biosecurity.govt.nz/files/regs/animal-welfare/pubs/naeac/animal-use-statistics.pdf

Above: ADI’sTechnical Briefingon the EU SeverityClassificationSystem.


PAIN RELIEF IN EXPERIMENTS

A paper in the journal Laboratory Animals recently cast ashadow over the use of analgesia in laboratory situations andits appropriate use in experimental animals.

The research assessed how frequently analgesia was used,which agents were used and determined if this use wasrelated to the severity of the procedure. The papers includedin the review covered thoracotomies, orthopaedic procedures,skin incisions, craniotomy, laparotomy and burn studies. Intotal, 149 papers were reviewed, 74 from 2000-2001 and 75from 2005-2006. The articles came from 61 different journals.

The paper highlighted worrying laxity inreporting procedures, which included:

� “dose was often given in mL/animal or

mg/animal rather than mg/kg”

� “In both time periods, duration of

systemic analgesia administration wasnot specified in 46% of papers thatreported the use of a systemicanalgesic”

� “When the administration of a systemic

analgesic was specified, the frequencyof systemic analgesic administrationwas not specified in the majority ofcases (68% of papers in both timeperiods)”

� “There was a wide reported dose

range of analgesics for manyspecies” – the authors explain how, inrabbits, with one compound, the dose“varied by 30-fold”, which “suggestsuncertainty about dose”

� Duration of analgesic administration

did not always increase with theseverity of the procedure. The authorsconcluded that “duration of analgesicadministration may often beinappropriate”

� Eight papers stated analgesia was

given “as necessary”, but nonedescribed how pain was assessed orthe signs that were used as indicatorsof pain.

In the discussion, the authors state that “given the sensitivity surrounding the use ofanimals in experiments, particularly when they involve non-human primates andcompanion species such as dogs, it is surprising that some peer-reviewed papers stilldo not report the administration of any form of analgesic.”

The paper states how “overall, rabbits, pigs, sheep, dogs and non-human primateswere more likely to receive analgesics following potentially painful experimentalprocedures than has been reported in laboratory rodents but analgesic administrationto ‘large’ laboratory species is still not optimal.”Coulter, C.A. et al (2009) “Reported analgesic administration to rabbits, pigs, sheep, dogs and non-human primates undergoingexperimental surgical procedures”, Laboratory Animals, vol.43, pp.232-238

Questions over painrelief in experiments

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Lord Dowding Fund for Humane Research www.ldf.org.ukUK: Millbank Tower, Millbank, LONDON, SW1P 4QP, UK. Tel: +44 (0)20 7630 3340

US: 953 Mission St., Suite 200, SAN FRANCISCO, CA 94103, USA. Tel: +1 415 543-2344

South America: Apartado Postal 359888 BOGOTÁ, Colombia.

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� Yes! I would like to support Lord Dowding Fund for Humane Research (LDF), and support medical and scientific research that helps people AND animals.

Please complete this form in BLOCK CAPITALS, using a ball point pen, and return by post to Lord Dowding Fund for Humane Research, Millbank Tower, Millbank, LONDON, SW1P 4QP, UK.

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Our research includes

The difference is ...

... we NEVER fund animal experiments

cancer

Alzheimer’s

vision

hearing

healingtoxicity testing

computer assisted learning

pain

Our research includes

The difference is ...

... we NEVER fund animal experimentsInvesting in the future with Good Science, Saving Animals and a Clear Conscience

cancer

Alzheimer’s

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hearing

healingtoxicity testing

computer assisted learning

pain

NS Magazine 2010

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