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					Number 12 - March 2002
NCA newsletter           Contents
Number 12 - March 2002   " EDITORIAL MARCH 2002                                                               1
                         " HUMAN ENDPOINTS REPLACE LETHAL PARAMETERS IN VACCINE QUALITY CONTROL               1
                         " ANTIGEN PRESENTING CELLS (APCS) GENERATED IN VITRO BY TRANSFECTING THE MHC
                            CLASS   II TRANSACTIVATOR – EVALUATED AS REPLACEMENT FOR APC ISOLATED FROM
                            THYMUS OR SPLEEN                                                                  5
                         " INVITOX 2002 - 12TH INTERNATIONAL WORKSHOP ON IN VITRO TOXICOLOGY                  7
                            FOURTH WORLD CONGRESS ON ALTERNATIVES AND ANIMAL USE IN THE LIFE SCIENCES         7
                         " MEETING REPORT
                            ADVANCING SCIENCE AND ELIMINATION OF THE USE OF LABORATORY ANIMALS FOR
                            DEVELOPMENT AND CONTROL OF VACCINES AND HORMONES.                                8
                         " OECD STOCKHOLM CONFERENCE REACHES AGREEMENT ON PRINCIPLES AND CRITERIA FOR
                            THE INTERNATIONAL REGULATORY USE OF TESTING METHODS                              11
                         " RUSSELL AND BURCH AWARD                                                           12
                         " NEW AALAS AWARD - THE BENNET J. COHEN ANIMAL STEWARDSHIP AWARD                    12
                         " ANNY ECK-HIEFF PRIZE                                                              13
                         " MEETING REPORT - ECVAM/ESTIV TEACHING WORKSHOP ON VALIDATION 13
                         " BOOK REVIEW: PAIN MANAGEMENT IN ANIMALS                                           14
                         " THE EUROPEAN RESOURCE CENTRE FOR ALTERNATIVES TO ANIMAL USE IN HIGHER
                            EDUCATION   (EURCA)                                                              15
                         " NEWSFLASHES                                                                       16
                         " DEVELOPMENT OF AN IN VITRO TEST BATTERY FOR THE ESTIMATION OF ACUTE HUMAN
                            SYSTEMIC TOXICITY;   OUTLINES OF THE EDIT PROJECT                                18
                         " THE DUTCH PROGRAMME ON ALTERNATIVES TO ANIMAL TESTING “DIERPROEVEN
                            BEGRENSD”                                                                        19
                         " OECD EXPERT PANEL RECOMMENDS REPLACING ANIMAL TESTS BY CELL AND TISSUE TESTS
                            FOR THE EVALUATION OF PHOTOTOXIC AND CORROSIVE PROPERTIES OF CHEMICALS   (BGVV
                            PRESS RELEASE)                                                                   19
                         " DE STICHTING BOUWSTENEN VOOR DIERENBESCHERMING ROEPT ONDERZOEKERS OP MEE
                            TE DINGEN NAAR DE    HUGO VAN POELGEEST-PRIJS ALTERNATIEVEN VOOR DIERPROEVEN 20
                         " NCA-THEMAMIDDAG ALTERNATIEVENDESKUNDIGEN                                       20
                         " FORTHCOMING EVENTS                                                                21
                         " COLOPHON                                                                          22
  Editorial March 2002                                                             Consequently, scientists have now
                                                                                   to struggle with the new term, used
In our today world, we have modern techniques available                            as they were to the former one.
to contact more people than ever before. Easy transporta-                          Also, when the animal welfare rep-
tion, phone, fax, e-mail and Internet have made it possible                        resentatives and scientist used the
to frequently exchange information with people from                                word replacement, it was clearly
different cultures and with different native languages.                            used in the context of the Three Rs.
                                                                                   When the legislators used the word,

T    o make these contacts success-
     ful we have to communicate.
One of the definitions of communi-
                                           tion problem became painfully
                                           obvious. At this meeting, scientists,
                                           regulators and animal welfare rep-
                                                                                   it was meant as replacing one test
                                                                                   by another, whether animal or in
                                                                                   vitro. Only after heavy discussions,
cation is ‘to give successful infor-       resentatives, from all over the         it became clear that different defini-
mation to others’. Here, successful        world, struggled to ‘communicate’       tions were applied.
is the key part of the definition. In      about the subject of the conference:    One of the benefits of meetings like
science, English has become the            “Develop, and achieve consensus         this is that in the end, when the
main language to exchange infor-           on, practical guidance on principles    meaning of most of the terms was
mation. Not always can we speak of         and processes for the validation and    agreed upon, the actual communi-
communication. English is not the          acceptance of animal and non-ani-       cation started. It also demonstrates
mother tongue of the authors of this       mal test methods for regulatory haz-    again, that the different back-
newsletter nor of some of the con-         ard assessment purposes”. To give       grounds of regulators and scientists
tributors. This may sometimes lead         some examples: The word “should”        hinder successful communication
to wrong communication, because            appeared to have slightly different     and as a result are a major hurdle in
either we use the wrong words to           meanings dependent on whether           the acceptation and implementation
inform you, or the words we use            the speaker was English or              of alternative methods in legisla-
may be correct in English, but might       American. In the UK, “should”           tion. More meetings like this would
have a slightly different meaning in       means “must”, in the US it means “it    improve the mutual understanding
US-English.                                would be better if”. A well-estab-      as well as the acceptation process
During the OECD conference on              lished term as “prediction model”       of alternative test methods.
“Validation and regulatory accept-         will now be named “data interpre-
ance of new and updated methods            tation procedure” since for some        Jan van der Valk
in hazard assessment” (Stockholm,          people not used to the earlier term,
6-8 March, 2002) the communica-            this covers the procedure best.



  Humane Endpoints Replace Lethal Para-
  meters in Vaccine Quality Control
The use of vaccines is one of the most effective preventive meas-
ures in fighting infectious diseases in human and veterinary med-
icine. During the development of vaccines but also of other bio-
logicals, a battery of tests has to be done. Besides analytical tests to
guarantee a consistent, validated production process, the safety
and potency of products have to be shown.


T    he production of immunologi-
     cal medicines is a biological
process and, therefore, is inherently
                                           cine is checked for quality by a
                                           panel of tests, most of them requir-
                                           ing the use of animals. The test bat-
variable. All the production               teries are legally stipulated by the
processes have to be strictly con-         European Pharmacopoeia (Ph.             mals are necessary for such tests.
trolled to ensure a safe, reliably effi-   Eur.) and by the relevant EU-,          For example, more than 120 mice
cacious and consistent vaccine.            WHO- and O.I.E.-guidelines.             are needed for potency tests for
Consequently, each batch of vac-           In practice large numbers of ani-       each batch of a diphtheria, tetanus,

                                                                                                          NCA - Newsletter - 1
pertussis, rabies and erysipelas vac-    tor of death. Furthermore, it should    • To organise conferences.
cine. Today, it is a legal require-      result in the greatest reduction in     • To prepare scientific papers and
ment in the EU that animals are          animal suffering, in both intensity       to communicate results e.g. by
treated with respect and only used       and duration.                             producing videos and CDs.
in research when it is absolutely        Humane endpoints should be iden-        • To act as a group of experts.
necessary. If animals have to be         tified and described when an
used, then two further criteria have     experiment is being planned, and        For the studies that will be reported
to be met. First, the minimum num-       be incorporated into the experi-        below, we decided to focus on
ber of animals should be used to         mental protocol.                        body weight, body temperature and
ensure statistical validity. Secondly,   Since challenge tests will continue     other clinical signs. These variables
the suffering of animals should be       to be used to estimate the potency      are often described in relation to
reduced to a minimum whilst still        of some vaccines for human and          models of infection and are practi-
meeting the scientific objectives.       veterinary applications, a Working      cable in routine potency tests.
This Three Rs approach –                 Group on Humane Endpoints for
Replacement,        Reduction     and    Lethal Parameters (HELP) was            Potency testing of whole cell
Refinement of RUSSELL & BURCH            founded with the support of             pertussis vaccine (Kendrick test).
(1959) – also applies to vaccine         ECVAM. HELP takes the task to           This test is routinely performed in
quality control.                         evaluate and validate the use of        the RIVM with an average annual
However, according to the Dutch          humane endpoints as an alternative      use of about 15.000 mice with 140
statistics on the use of laboratory      to severe clinical and lethal end-      animals per test. Animals are immu-
animals, about 16% of all animals        points in vaccine potency tests. In     nised and 14 days thereafter intra-
used in the production and quality       addition, HELP promotes the imple-      cerebrally challenged with virulent
control of biologicals, suffered         mentation and use of humane end-        Bordetella pertussis. Animals are
severe pain or distress due to the       points in guidelines for laboratory     observed for a period of 14 days.
test procedures. Examples of tests       staff carrying out vaccine quality      Data on surviving animals per vac-
that induce high levels of pain and      control tests.                          cine dilution group are used to cal-
suffering are the potency tests          The working practices of HELP are       culate potency by probit analysis.
based on a challenge procedure.          as follows:                             The body temperature was telemet-
One particular approach to reduce        • To perform an enquiry at control      rically determined with a subcuta-
animal pain and suffering is the             laboratories in order to gain an    neously implanted temperature-sen-
introduction of humane endpoints             insight into the endpoints that     sitive microchip transponder. In
as an alternative – one of                   can be used and to obtain infor-    addition, animal’s body weight was
Refinement. A humane endpoint                mation on studies using humane      recorded daily. Based on these
can be defined as the earliest indi-         endpoints.                          parameters, clinical score sheets
cator in an animal of pain, distress,    • To evaluate the use of non-lethal     were prepared for individual ani-
suffering, or impending death. This          endpoints in the potency test on    mals.
can then be used to terminate expo-          a limited number of products.       First, clinical signs of the disease in
sure and possibly start therapy, thus    • To identify and validate poten-       mice were defined in four different
allowing the animal to be humane-            tial humane endpoints.              stages. Stage 1 is characterised by
ly killed or to recover as soon as the   • To make recommendations for           less alert animals, their fur becomes
scientific objective has been                codes of best practice in chal-     dull and pilo-erection, especially
achieved (CCAC). Ideally, the end-           lenge tests (frequency of obser-    around the neck can be seen. The
point should be easy to monitor, be          vations, humane endpoints,          animals show a hunched back pos-
reproducible, and be a valid predic-         humane killing etc.)                ture and pilo-erection over the

Figure 1. Disease stages of pertussis infection in mice




              Stage 1                    Stage 2                       Stage 3                       Stage 4

NCA - Newsletter - 2
entire body during Stage 2. In addi-
tion, they become inactive and
socially isolated, and have little or
no food and water intake, resulting
in dehydration and emaciation.
The cardinal clinical signs in Stage 3
are the same as for Stage 2, but ani-
mals also show disturbed locomotor
activity. Animals fall easily after
being pushed, but are still able to
get up again. In Stage 4 the disease
climaxes in tonic-clonic convul-
sions, the animals become coma-
tose and finally die.
Conclusions from the study: A body
temperature decrease below 34,5 °C       Figure 2
and the clinical signs described in      * normalised to day two after infection
stage 3 can be used as surrogate         G: bodyweight; L: body temperature;       : score of clinical signs; shaded area: weight loss
endpoints to replace lethality. Body     > 15%
weight could decrease significantly
during the disease but was not a         voked. By this time, there also is a            humane endpoints reduces each
good indicator for predicting            marked loss of body weight. In                  animal’s suffering by about 3 – 5
impending death. Introducing sur-        Stage 4, lameness, and paresis, usu-            days.
rogate humane endpoints reduces          ally of the hind legs, soon go on to
each animal suffering by about 0.5       complete paralysis and animals                  Efficacy testing of swine erysi-
– 2 days.                                become clearly dehydrated. In                   pelas vaccines
                                         Stage 5 animals become moribund                 During licensing procedures, the
Batch potency tests of rabies            and are prostrate and recumbent                 efficacy of vaccines has to be
vaccines (NIH-test)                      and so they obviously do not feed               shown in the target species, which
Mice are vaccinated intraperitoneal-     or drink and yet they may still sur-            is the pig for erysipelas vaccines.
ly with several vaccine dilutions.       vive for one or two days. All mice              The test conditions and the chal-
Two weeks later, they are chal-          that showed Stage 3 never recov-                lenge cultures are standardised and
lenged with an intracerebral injec-      ered and went on to die some days               have been evaluated in several
tion of rabies virus under anaesthe-     later.                                          laboratories. Thus, no additional
sia. Overall, the rabies infection in    Body weight loss is a very early                tests should be necessary to estab-
mice is characterised by a slow          indicator of an infection, being                lish humane endpoints. Serovars 1
onset of disease usually beginning       noted even before other clinical                and 2 are available as European
between 4 and 6 days after infec-        signs were evident. Weight loss pro-            Biological Reference Preparations,
tion.                                    gressed continuously and could                  which have been tested in several
Typical clinical signs of rabies in      reach between 30 – 50 % shortly                 laboratories. This showed that it is
mice showed up in five stages with       before death. A loss of body weight             possible to test protection against
healthy animals in Stage 0. Ruffled      of > 20 % alone was also a suitable             several serovars simultaneously in
fur and a hunched back in Stage 1        humane endpoint.                                the same pig, thereby reducing the
reflect that the animal’s welfare is     Conclusions from the study: A                   number of animals required.
already compromised. During Stage        decrease in body weight or the                  The test requires 15 pigs of the
2, animals lose their alertness, and     appearance of typical signs of neu-             same origin more than 12 weeks
walk more slowly than usual. They        rological disorder allows terminat-             old and free from erysipelas anti-
may show circling movements              ing the experiment without any loss             bodies. Ten pigs are vaccinated
mainly in one direction. In Stage 3      of scientific data. This saves the ani-         with the vaccine under test and five
the neurological signs become            mals a slow progressive death.                  are used as unvaccinated controls.
increasingly obvious with trembling      Combining these two points allows               Three weeks after vaccination, the
and shaky movements, and convul-         an experiment to be terminated at               pigs are challenged with virulent
sions appear and can often be pro-       an even earlier phase. Introducing              strains of Serovars 1 and 2 of

                                                                                                                    NCA - Newsletter - 3
Erysipelothrix rhusiopathiae. At            The infection may kill the pigs if       • Treatment should begin as soon
least 4 out of the 5 control animals        they do not receive treatment. The         as possible after typical clinical
must show typical signs of infection        bacterium is very sensitive to antibi-     signs have developed to ensure
with erysipelas, and at least 9 out of      otic and antiserum treatment.              that unnecessary suffering is
10 vaccinated animals have to be            Therefore, therapeutic intervention        avoided. Effectiveness of treat-
protected and must not show any             at an early stage can avoid unnec-         ment will be seen as the animals
signs of disease.                           essary suffering and death. The clin-      showing normal behaviour and
A challenge is performed by an              ical signs of erysipelas can be easi-      feeding the next day, but it may
intra-dermal injection of 0.1 ml of a       ly monitored using score sheets that       take several days for the skin
bacterial suspension. This method           document the clinical signs of skin        lesions to heal and disappear.
induces typical pathognomic signs           lesions and fever.                         Using this scheme and treat-
to develop at the inoculation site.         Treatment or humane killing should         ment, no death has been report-
Thus, the development of infection          be initiated in the following circum-      ed so far.
can easily be monitored and con-            stances.
trolled. During the following days,         • Reaction at the skin injection site    We hope that many institutions and
the lesions develop in unprotected              together with high body temper-      companies will come to use such
animals showing a typical brick- or             ature (> 2°C above baseline, or      humane endpoints instead of lethal-
diamond-shaped appearance (“dia-                > 41.0 °C).                          ity. Our films and CDs may help to
mond skin disease”).                        • Very high fever reaction (> 41.0       show that it is possible, and impor-
Monitoring body temperature                     °C), anorexia and recumbency         tant to practise humane science,
should start a few days before vac-             without skin reaction. In these      thus preventing avoidable animal
cination to determine their baseline            cases, blood samples should be       suffering.
temperature and for the animals to              taken for culture to demonstrate
become used to this procedure.                  bacteraemia, followed by treat-      The general idea of HELP’s activity,
Monitoring should continue for at               ment of the animals.                 the application of humane end-
least 8 days after challenge and be         • Antibiotic treatment should also       points, has been considered by the
measured twice daily. All unvacci-              be given (e.g. long-acting peni-     European Pharmacopoeia. The
nated pigs challenged with these                cillin for a minimum of five         revised monograph “Vaccines for
strains developed typical signs of              days) to all unprotected animals     human use” (Ph.Eur. 4th ed, Suppl.
acute erysipelas (high fever > 41.0             at the end of the test observation   2002) and the draft revision propos-
°C with and without local skin reac-            period     to     avoid   chronic    al of the general monograph for
tion). Protected animals show no                erysipelas lesions such as endo-     veterinary vaccines now includes a
significant increase in temperature.            carditis or arthritis.               new section on Humane Endpoints
                                                                                     which reads:
Figure 3                                                                             “Animal tests. In accordance with
Score sheet for Erysipelas Efficacy tests                                            the provisions of the European
                                                                                     Convention for the Protection of
                                                                                     Vertebrate Animals Used for
                                                                                     Experimental and Other Scientific
                                                                                     Purposes, tests must be carried out
                                                                                     in such a way as to use the mini-
                                                                                     mum number of animals and to
                                                                                     cause the least pain, suffering, dis-
                                                                                     tress or lasting harm. The criteria
                                                                                     for judging tests in monographs
                                                                                     must be applied in the light of this.
                                                                                     For example, as soon as there is suf-
                                                                                     ficient indication of a positive result
                                                                                     e.g. typical clinical signs or pending
                                                                                     death, then an animal should be
                                                                                     either humanely killed or given suit-
                                                                                     able treatment to prevent avoidable
                                                                                     suffering”.

NCA - Newsletter - 4
More information about these stud-       (http://www.lal.org.uk/cont.htm)         P.O. Box 1, 3720 BA Bilthoven,
ies and also on the use of humane        or from the secretariat of HELP:         The Netherlands,
endpoints in other areas can be          c/o Björn Steen,                         Phone: +31 / 30 / 27 42 377,
found in the Proceedings from the        National Institute of Public Health      Fax: +31 / 30 / 27 44 408,
international conference “Humane         and the Environment                      e-mail: Bjorn.Steen@rivm.nl.
Endpoints in Animal Experiments          (RIVM, Central Animal Labora-
for Biomedical Research 1998” in         tories),
Zeist



  Antigen presenting cells (APCs) generated in vitro by
  transfecting the MHC class II transactivator – evaluated
  as replacement for APC isolated from thymus or spleen

Antigen-specific T cell lines have become an important object and tool for immunological
research. They have to be continuously propagated in vitro. This is done in cycles of
restimulation with antigen presenting cells (APCs) displaying the respective antigen to T
cells via the MHC molecule to which the T cells are restricted.


T    his procedure requires the iso-
     lation of APCs from thymus or
spleen originating from animals of
                                         cessfully transfected with a vector
                                         (EBO-76PL; Peijnenburg et al. 1997)
                                         containing the human CIITA gene.
                                                                                  cell-subpopulation was noticed.
                                                                                  Therefore, MHC class II expression
                                                                                  was induced by rat IFN-γ. As CIITA
the same syngeneic strain as the T       A subpopulation of Rat-2 cells trans-    is the key-element of the MHC class
cells. This need for animals as APC-     fected with the CIITA vector dis-        II induction pathway which is
source could be overcome if con-         played cell surface expression of        induced by IFN-γ, this procedure
tinuously growing cell lines with        MHC class II molecules, when ana-        mimicked the maximal class II
APC-quality were available. Such         lyzed with anti-MHC II antibodies        expression that was to be expected
cell lines do not exist for most         by FACS. Thus it could be demon-         by CIITA-transfected cells. Under
strains of laboratory animals. Our       strated for the first time, that the     these conditions an overall low
proposal encompassed the genera-         human CIITA, most likely due its         class II expression was observed.
tion of in vitro cultured rat APC-       interspecies conservation, was able      The same low MHC class II expres-
lines by introducing the gene for        to induce MHC class II expression        sion was also observed after IFN-γ
the transcription-factor MHC class II    in rat cells.                            stimulation of another MHC II
trans-activator (CIITA) to cell lines.   To ensure that the vector was not        expressing fibroblast-like rat cell
This was developed in collabora-         lost during cell proliferation, trans-   line, the astrocyte line F10 (a kind
tion with Peter van den Elsen            fected cells were selected by            gift of Prof. Dr. Wekerle, University
(Leiden University) who provided         Hygromycin B, an antibiotic for          Würzburg, Germany).
us with the vector containing the        eukaryotic cells for which a resis-
human CIITA-cDNA (Steimle et al.,        tance gene was present on the            We conclude from these experi-
1994). This transcription-factor         CIITA-vector. Cultures of CIITA          ments that, although MHC class II
induces the expression of all iso-       transfectants were sorted by FACS        was inducible by CIITA, levels of
types of MHC class II molecules and      according to high MHC class II           expression by the rat cell lines
some other molecules necessary for       expression. The sorted cells, still in   available, in comparison to ex vivo
antigen presentation (Chang and          the presence of selection medium,        APCs like macrophages and den-
Flavell, 1995).                          were expanded but lost their MHC         dritic cells, are dependent on other
To obtain functional APC the MHC         class II expression after a view         (genetic) factors, probably associ-
class II negative rat fibroblast cell    cycles of proliferation for unknown      ated with their differentiation as
line Rat-2 (Topp, 1981; provided by      reasons.                                 fibroblast-like cells.
the European Collection of Cell          A general low expression of MHC
Cultures in Salisbury, UK) was suc-      class II molecules by the transfected    The crucial question was whether

                                                                                                        NCA - Newsletter - 5
                                                                                       the cell transforming T antigens of
                                                                                       SV40 (simian virus).

                                                                                       Conclusions
                                                                                       The human CIITA is functional in
                                                                                       rat cells and CIITA-induced MHC
                                                                                       class II expression is cell (line)
                                                                                       dependent. The CIITA-transfected
                                                                                       fibroblast-like cell lines available
                                                                                       are able to present peptide-antigen
                                                                                       to T cells, their function as APC is,
                                                                                       however, not optimal. This should
                                                                                       be solved by using continuously
                                                                                       growing cell lines of professional
Fig.1:                                                                                 APC-background with constitutive
F10 cells were stimulated with 500 U/ml rat IFN-γ for 48h before they were tested.     MHC class II-expression induced by
Stimulation of the A2b-hybridoma was measured by IL-2 dependent proliferation of the   CIITA transfection.
CTTL-16 cell line.
                                                                                       References
the MHC class II positive cells were        fibroblast-like type, are not suited to    Chang, Flavell, R.A. 1995. Class II trans-
                                                                                          activator regulates the expression of
able to function as T cell stimulating      optimally stimulate T cells even              multiple genes involved in antigen
APCs. IFN-γ-stimulated cell lines           when rendered MHC class II posi-              presentation. J.Exp. Med. 181:765.
(Rat-2 and F10) were thus tested for        tive, most probably due to a low
                                                                                       Fukomoto, I., McMaster, W.R., Williams,
their antigen presentation capacity         level of MHC class II expression. In          A.F. 1985. Mouse monoclonal anti-
towards a well defined T cell clone         other words, target cell lines for            bodies against rat major histocom-
                                                                                          patibility antigens. Two Ia antigens
(A2b) for which the antigen (hsp60)         transfection were not capable of
                                                                                          and expression of Ia and class I
was known and could be added in             inducing extensive T cell stimula-            antigens in rat thymus. Eur. J.
the form of a synthetic peptide. In         tion. Thus, these transfected cells           Immunol. 12:237.
other words presentation of this            would not be suitable for the opti-
                                                                                       Peijnenburg, A., Gobin, S.J.P., Van
peptide to T cells would not require        mal in vitro expansion of CD4+ T               Eggermond, M.C.J.A., Godthelp,
extensive      antigen     processing       cells. It would therefore be neces-            B.C., Van Graafeiland, N. Van den
                                                                                           Elsen, P.J. 1997. Introduction of
events. Despite this, the cells             sary to obtain or generate continu-
                                                                                           exogenous class II trans-activator in
proved to be unable, in contrast to         ously growing cell lines of profes-            MHC class II-deficient ABI fibrob-
thymic or splenic APC, to stimulate         sional APC like macrophages or                 lasts results in incomplete rescue of
                                                                                           MHC class II antigen expression. J.
a proliferative response of the T cell      dendritic cells. Cell lines of
                                                                                           Immunol. 159:2720-2727.
clone. As the level of co-stimula-          macrophages, which are not consti-
tion, provided by the cell lines            tutively class II positive would then      Steimle, V., Siegrist, C.A., Mottet, A.,
                                                                                           Lisowska-Grospierre, B. Mach, B.
could be responsible for this, a co-        be rendered class II positive by               1994. Regulation of MHC class II
stimulation-independent T cell              transfection with CIITA. As profes-            expression by Interferon-g mediated
hybridoma (A2b-hybridoma) was               sional APC represent a final stage of          by the transactivator gene CIITA.
                                                                                           Science 265:106.
tested for stimulation (IL-2 produc-        cell differentiation, they are either
tion) in response to antigen-pulsed         unable to proliferate (dendritic           Topp, W.C. 1981. Normal rat cell lines
IFN-γ-stimulated Rat-2 and F10              cells), or do so only for a few cell          deficient in nuclear thymidine
                                                                                          kinase. Virology 113:408.
cells. Now positive T cell stimula-         cycles (macrophages) upon isola-
tions were obtained. However, in            tion and in vitro culture. This can        U. Wendling and W. van Eden
comparison to ex vivo isolated APC,         be overcome by immortalisation             Department of Immunology
the IFN-stimulated cell lines stimu-        and transformation of such cells           Institute of Infectious Diseases and
lated the hybridoma to a 100-fold           starting from primary cell cultures.       Immunology
lower degree (see Fig. 1).                  This might be accomplished by set-         Faculty of Veterinary Medicine
                                            ting up a transfection system for cell     Utrecht University
The findings described above lead           transformation, for example by             Yalelaan 1
us to the conclusion, that the rat cell     employing a vector (pSV3-neo;              3584 CL Utrecht
lines available, which are of a             ATCC code: 37150) that comprises           The Netherlands

NCA - Newsletter - 6
                 INVITOX 2002
  12   th
            International Workshop on In
                Vitro Toxicology

   Golfo di Gaeta Centro CONI – Formia, Italy
               16-19 October 2002

INVITOX is an international workshop held in Europe
every two years since 1980. It is organised by ESTIV,
the European Society of Toxicology In Vitro, with the
aim to promote exchange of views, information and
experience among scientists involved in the develop-      Fourth World Congress on
ment and use of in vitro methods in the area of toxi-
cology.                                                  Alternatives and Animal Use
Scientific Program                                            in the Life Sciences
The program will consist of eight main sessions
including invited lectures, oral communications
(selected from the submitted abstracts) and poster
presentations:
                                                         New Orleans, Louisiana, USA,
• Biomarkers of exposure and damage: environ-
  mental pollutants.                                                    11-15 August
• Advanced models for toxicity studies on differen-
  tiating cells.
• Interindividual variability in pharmaco-genetics.
                                                         The Congress will be organized around five themes:
• Inflammation mechanisms and signal transduc-
  tion in in vitro studies.                                  A. Replacement and Reduction Alternatives
                                                                 Chairs or Organizing Committee: Dr. Jose
• Proteomics and genomics and High-throughput
                                                                 Castell (SPAIN) and Dr. June Bradlaw (USA)
  screens in in vitro toxicology.                            B. Policy and Ethics
• Role of in vitro systems in food safety: tradition-            Chair or Organizing Committee: Dr. Jon
  al and novel food or food ingredients.                         Richmond (UK)
• Human cells in vitro: sources, safety and ethical          C. Refinement and Reduction Alternatives
                                                                 Chairs or Organizing Committee: Dr. David
  issues.
                                                                 Morton (UK)
• Regulatory issues: National Platforms, Cosmetic            D. Education and Information Resources
  testing. Integrated testing strategy.                          Chairs or Organizing Committee: Dr. Barbara
                                                                 Grune (GERMANY) and Nicole Duffee (USA)
Website                                                      E. Test Development, Validation and
Bookmark the INVITOX 2002 website and keep up to                 Implementation
date with all the latest developments. Direct e-mail             Chairs or Organizing Committee: Dr. William
                                                                 Stokes (USA) and Dr. Julia Fentem (UK)
links to the conference organisers will ensure quick
and easy submission of abstracts and registration
(www.xs4all.nl/~shorbach/INVITOX)                        For more information visit the congress website at:
                                                         http://www.worldcongress.net/
Registration
                                                         Or contact the organisation:
The number of participants is restricted to 150. The
                                                         The Alternatives Congress Trust
registration fee will be in the range of 700-800 euros   700 Professional Drive
and will include attendance at the scientific and        Gaithersburg, MD 20879
social events, abstract booklet, all meals, accommo-     Tel: (301) 548-7771
dation (4 nights), workshop proceedings (to be           Fax: (301) 548-7726
published in a special issue of Toxicology In Vitro),    E-mail: fourthworldcongress@starpower.net
and taxes. Discount rates will be applied for ESTIV
members and students. A limited number of bursaries
will be available for young scientists. Details can be
obtained at the e-mail address:
invitox@iss.it

                                                                                               NCA - Newsletter - 7
  MEETING REPORT
  Advancing Science and Elimination of the Use of Laboratory
  Animals for Development and Control of Vaccines and Hormones.
  Jaarbeurs Congres Centre, Utrecht, The Netherlands, 12-14
  November 2001


U     nder the auspices of the
      International Association for
Biologicals (IABs), the National
                                          monitoring Three Rs progress and
                                          giving guidance to future activities.
                                          The opening speech was given by
                                                                                  Commission, the OMCLs network
                                                                                  and the relation with the licensing
                                                                                  authorities. Dr.J. Milstien (WHO,
Institute of Public Health and the        Prof. Michael Balls (ECVAM). He         CH) addressed the change in the
Environment (RIVM, NL), the Paul-         addressed the progress in applying      role of the National Control labora-
Ehrlich-Institut (PEI, D) and the         the Three Rs of Russell & Burch to      tory, giving stronger dependence to
National Institute for Biological         the testing of biological products.     consistency in production, to GMP
Standards and Control (NIBSC, UK)         As a point of reference, Prof. Balls    and to experience in the clinic. The
a symposium on “Advancing                 used the ECVAM workshop on              role of the animal tests, with their
Science and Elimination of the Use        immunobiologicals that was held in      intrinsic variability and failure in
of     Laboratory     Animals       for   1994. He concluded that progress        some cases to provide an appropri-
Development and Control of                has been impressive, but also           ate model system for the human
Vaccines and Hormones” was held           addressed some points of concern,       response, should be limited to con-
at the Jaarbeurs Congres Centre,          such as the re-testing policy by        firmation of consistency. The first
Utrecht (NL), from 12-14 November         Official    Medicinal       Control     scientific session was on “The Three
2001. The objectives of                                                                        Rs and Safety Testing:
the symposium were to                                                                          current activities”. In his
present an overview of                                                                         key-note lecture K.
Three Rs progress being                                                                        Cussler (PEI, D) present-
made in the field of the                                                                       ed an overview of
production and quality                                                                         opportunities for animal
control of human and                                                                           replacement and reduc-
veterinary       biological                                                                    tion of toxicity tests. In
products and to offer a                                                                        particular, he addressed
forum for an exchange of                                                                       the discussions about
information.        Special                                                                    general safety tests, such
attention was given to                                                                         as the target animal safe-
validation procedures; one of the         Laboratories             (OMCLs).       ty test for veterinary vaccines and
conference sessions focussed on           Representatives of the European         the abnormal toxicity test for
progress reports of ongoing valida-       Department for the Quality of           human vaccines. He stressed the
tion studies.                             Medicines (EDQM), the United            need for harmonisation of require-
The meeting was attended by more          States Department of Agriculture,       ments. Interesting presentations
than 150 participants from 34 differ-     Center for Veterinary Biologicals       were given by O. Ottiger (IVI, CH)
ent countries. The programme              (CVB) and of the World Health           and B. Kegel (PEI, D) on the PCR
included 9 sessions with a total of       Organization (WHO) discussed the        technology for the detection of
44 presentations, 7 parallel break-       Three Rs in the regulatory process      extraneous agents in avian viral
out groups and 22 posters on dis-         context. A. Artiges, Director of        vaccines and on quantification of
play. Prof. van der Zeijst, director of   EDQM, related the Three Rs activi-      proteolytic activity of tetanus toxin
the vaccine division of RIVM wel-         ties to the various components of       as a replacement alternative for spe-
comed the participants. He empha-         EDQM; the biological standardisa-       cific toxicity of tetanus vaccines,
sised the positive contribution that      tion programme, the policy of the       respectively. The session on
these types of conferences make to        European            Pharmacopoeia       “Potency Testing and the Three Rs:

NCA - Newsletter - 8
current activities” was introduced       Evaluation of 14 batches of EPO           several organisations to the imple-
by M. Schwanig (PEI, D). He sum-         showed that the N-Glycan charge           mentation of Three Rs alternatives
marised our efforts in the past years    assay is a potential alternative to the   was the theme of session 6. From
by making a division in two kind of      exhypotic polycythemic mouse              an industry perspective (M. Chapek,
approaches: a) replacement of the        bioassay. The evaluation of the           MVP lab, USA) it seems that many
challenge procedure in potency           capillary zone electrophoresis for        of the major obstacles and their
testing by a serological determina-      potency testing of FSH revealed a         potential solutions involve govern-
tion and b) a reduction of the num-      good correlation with the in vivo         mental requirements. Although in
bers of animals by going from a          ovarian weight augmentation test.         vitro assays are theoretically more
multi- to a single dilution assay.       Session 4 discussed the group of          cost effective and timely than in
Several issues were discussed in the     “Other Biologicals”, in particular        vivo test, regulations may eliminate
following session presentations: the     therapeutic toxins and allergens. In      these advantages, making current
reduction of the numbers of ani-         her key-note lecture, D. Sesardic         technology less attractive to manu-
mals in potency testing by applica-      (NIBSC, UK) addressed the in vitro        facturers of biologicals. Some key
tion of a non-linear mixed effect        biochemical assays for clostridial        issues and problematics encoun-
(NLME) model for parallel line           neurotoxins. K. Kaul (PEI, D) pre-        tered from the view of the regulator
analysis (K. Haslov, SSI, D); an in      sented data of replacement of             were addressed by J-M. Spieser
vitro potency test for tetanus vac-      murine sera by allergan-specific          (EDQM, F). J.L. Di Fabio (PAHO,
cines based on antigen quantifica-       monoclonal IgE antibodies.                USA) summarised the possibilities
tion by ELISA detection of specific      The state-of-the art of seven ongo-       and barriers of Three Rs alternatives
Hc fragment epitopes (S. Prieur,         ing validation studies were sum-          implementation in developing
AFSSPS, F); a combined test for          marised in Session 5. These includ-       countries. For example, the possible
assessment of potency and specific       ed: the serological methods for           higher costs of alternative tech-
toxicity of whole cell pertussis vac-    potency testing of tetanus toxoid         niques have been suggested as a
cine (v. Straaten, RIVM, NL) and an      vaccines (C. Hendriksen, RIVM, NL)        major drawback for their implemen-
enzymatic-HPLC to measure the            and diphtheria toxoid vaccines            tation. The ECVAM guidelines on
enzymatic ribosylation activity of       (Winsnes, NMA, N); the prevalida-         prevalidation and validation were
active pertussis toxin as a replace-     tion of physicochemical methods           presented by M. Halder (ECVAM, I).
ment for the current in vivo hista-      for FSH potency (Storring, NIBSC,         These guidelines were illustrated
mine sensitisation test (HIST) (D.       UK); batch potency testing of inac-       with lessons learned from previous
Xing, NIBSC, UK). The third session      tivated erysipelas vaccines by ELISA      ECVAM validation studies. The last
focussed on Testing of Hormones.         (Rosskopf, PEI, D); an in vitro anti-     lecture of session 6 was given by G.
Substantial progress has been made       gen ELISA for inactivated Newcastle       Calver      (BBR,    Canada).     He
in this field. In his key-note lecture   Disease vaccines (Claassen, ID-           addressed some of the activities of
A. Bristow (NIBSC, UK) summarised        Lelystad, NL); the establishment of       the Bureau of Biologicals, such as
three approaches: adoption of bio-       European Pharmacopoeia biologi-           the evaluation of humane end-
identity tests using fewer animals,      cal reference preparations for            points. Session 7 highlighted the
replacement of in vivo assays with       potency testing of clostridial vac-       use of serum free media. The key-
in vitro assays, and replacement of      cines (Esposito-Farese, EDQM, F)          note speaker, O-W. Merten
bioassays with physico-chemical          and in vitro pyrogen tests (Hartung,      (Genethon III, F) discussed the
assays. However, for one class of        STZInPuT, D). Key-words coming            drawbacks associated with the use
therapeutic hormones, the glyco-         up from these presentations are:          of serum or animal derived sub-
proteins, this process has proved to     high commitment, involvement of           stances such as the potential intro-
be problematic, due to heterogenic-      regulatory authorities, substantial       duction of adventitious agents into
ity at the molecular level and the       need for human and financial              the final product. However, he indi-
presence of the terminal sialic acid     resources and promising results           cated that serum free media,
which has in vivo activity but little    coming from these studies.                although more defined, still might
receptor binding and signal trans-       However, it also became clear that        be a risk for the introduction of
duction in vitro. Some practical         the need for human and financial          viruses as serum free media are
examples of Three Rs approaches          resources is substantial, that there is   generally supplemented with puri-
were presented by P. Hermentin           only slow progress and that lack of       fied proteins, peptones, etc. mainly
(Aventis Behring, D) and J. Mulders      harmonisation is an obstacle that         from animal or human origin. In
(Organon,        NL),    respectively.   shouldn’t be ignored. The view of         this context, the use of substances

                                                                                                         NCA - Newsletter - 9
of non-animal origin (e.g. plant ori-   Further new quality control tech-        Three Rs methods in 3RD World
gin) might be of interest. Further      niques were presented in Session 9.      Countries. In the closing ceremony,
presentations in this session were      A particular important aspect is the     J. Löwer (PEI, D) stressed the need
on the standardisation and charac-      application of biochemical and           for an open dialogue, high commit-
terisation of animal cell lines (G.     physicochemical tests, such as NMR       ment to the Three Rs and common
Stacey, NIBSC, UK) and on               technology, to characterise vaccine      sense in the acceptance of Three Rs
advanced process monitoring (R.         antigen and formulations. In his         alternatives. In that regard, the fol-
Neeleman, LUW, NL). The full title      key-note lecture, R. Sitrin (Merck,      lowing statement of J. Di Fabio
of the last presentation was “the       USA) illustrated this approach for       (PAHO, USA) should be cited:
ultimate alternative”. According to     polyvalent pneumococcal vaccine.         “Examination of the way in which
the speaker’s opinion, in process       The use of in vitro functional tests,    animal-use intensive procedures
monitoring using in vitro methods       such as stimulation of cytokine-pro-     became ‘gold standards’, reveal lim-
provides the best assurance of con-     ducing cells, or in vitro antibody       ited validation in the modern sense.
sistency and quality and, therefore,    production were illustrated by K.        Therefore we propose that common
offers the opportunity to complete-     McCullough (IVI, CH). He empha-          sense and application of the best
ly eliminate the use of animals for     sised the need for intercellular sig-    science available, and not tradition
batch release and regulatory            nals by co-culture cells of the          alone, should be the basis for
approval. New approaches in vac-        immune systems with endothelial          deciding on the use of alternative
cine development and quality con-       cells. The immune response is            assays”.
trol were addressed in session 8. In    based on a very complex process.         Abstracts and most of the confer-
his key-note lecture D. Wood            For the time being, the application      ence power-point presentations are
(WHO, CH) discussed new vaccine         of in vitro methods will have two        available on the meeting web-site:
technologies, based on new science      forms. Firstly, to improve the design    www.rivm.nl/iabssymp2000/
(genomics;      proteomics),    new     and efficacy of subsequent in vivo
approaches (new combination vac-        experiments, and secondly, obser-        C.F.M. Hendriksen
cines; new delivery options) and        vations from in vivo experimenta-        National Institute of Public Health
new needs (changes to substrates        tion can be studied further by in        and the Environment (RIVM)
or formulations) and the effects on     vitro means. In the last three pre-      P.O.Box 1
the use of animals. His conclusion      sentations, some examples of novel       3720 BA Bilthoven
was rather ambiguous. The extent        technologies were given. B. Hallis       The Netherlands
of usage may vary depending on          (CAMR, UK) discussed the use of in
whether a new vaccine is based on       vitro functional assays for the deter-   Netherlands Centre on Alternatives
an entirely new concept or drawn        mination of anthrax toxin compo-         to Animal Use (NCA)
on prior experience with a particu-     nents, T. Ranheim (Merck, USA)           Utrecht University
lar antigen. J. Kreeftenberg (RIVM,     presented the RT-QPCR technology         Yalelaan 17
NL) clarified the WHO proposal to       for potency testing of live vaccines     3584 CL Utrecht
harmonise current test methods for      and T. Leung (NIBSC, UK)                 The Netherlands
quality control of toxoid vaccines      addressed the application of an in
into simplified consistency tests.      vitro endopeptidase assay for detec-     K. Cussler
Essential in this approach is the       tion of residual tetanus toxin activi-   Paul-Ehrlich-Institut
demonstration of batch-to-batch         ty in tetanus toxoid.                    Paul-Ehrlich-Strasse 51-59
consistency in biochemical and          On the second day of the confer-         63225 Langen
immunological characteristics. The      ence, seven parallel breakout group      Germany
WHO proposal is to use vaccine lot      sessions were held, given the
that has been shown to be clinical-     opportunity for each participant to
ly safe and effective. Practical        attend two of these workshops.
examples of in vitro procedures         Breakout group topics included:
with the potential to replace the in    new vaccine production technolo-
vivo potency test were given by T.      gies, the consistency approach, ani-
Schofield (Merck, USA) for recom-       mal care best practices, validation
binant hepatitis B vaccine and by R.    strategies, safety testing, the inter-
Maas (ID-Lelystad, NL) for inactivat-   play between regulator, licensing
ed Newcastle disease vaccines.          authority and manufacturer and

NCA - Newsletter - 10
  OECD Stockholm Conference reaches agreement on principles and
  criteria for the international regulatory use of testing methods

Experts from OECD coun-                  ance on the description of a new        3 R’s principles (Replacement,
tries, including govern-                 benchmark for the quality of testing    Reduction and Refinement of ani-
ment, academia, industry                 methods.       This so-called Test      mal tests) in method development.
and animal welfare repre-                Prediction Description defines the
sentatives, have agreed on               steps to be taken to convert results    A detailed Report of the Conference
fundamental       scientific             from the test method into a predic-     recommendations will be made
principles and practical                 tion of the hazard that is useful for   available in May/June this year.
criteria for the regulatory              making regulatory decisions. For        Subsequently,      a      Guidance
acceptance of test methods               example: for each cell culture (non-    Document for the regulatory
used for safety assessment               animal) test a scientific description   acceptance of test methods for haz-
of chemical substances and               is needed to explain what an effect     ard assessment will be finalised,
products.                                observed in the cell culture would      taking into account the agreements
                                         predict in terms of human health if     reached at the Conference. The

A     Conference was held from 6-8
      March in Stockholm, Sweden
to agree on practical guidance for
                                         human health would be the test’s
                                         target. But a similar justification
                                         would be required for each animal
                                                                                 Guidance Document, once formally
                                                                                 adopted by OECD’s Member coun-
                                                                                 tries, will be used as the interna-
assessing the reliability and rele-      test to relate changes observed in      tional consensus document for the
vance of animal and non-animal           animals after chemical exposure to      validation and regulatory accep-
testing methods for regulatory haz-      a prediction of what may be expect-     tance of new methods in hazard
ard assessment purposes.         The     ed in man after similar exposures.      assessment.
Conference was a follow-up to the                                                The OECD website on the Test
1996 OECD Workshop of Solna,             The Conference further agreed on        Guidelines Programme
Sweden where agreement was               the need for and the details of an      (http://www.oecd.org/ehs/test) will
reached on the principles for vali-      independent review process to con-      provide further details of the
dation of new testing methods. At        firm the quality of each validation     Conference and its outcome.
that time it was already recognised      study and facilitate the regulatory     Inquiries can also be made to
that in addition to these principles,    use of new methods in hazard            Herman B.W.M.Koëter, Principal
policies and guidance should be          assessment. Although independent        Administrator of the OECD Test
developed to facilitate their applica-   peer reviews are common practice        Guidelines Programme
tion.                                    for acceptance of publications in       (herman.koeter@oecd.org).
                                         the scientific literature, structured
Approximately 100 experts from 15        and transparent independent peer
Member countries, the European           reviews are currently only marginal-
Commission, industry and animal          ly applied in the acceptance
welfare groups participated in the       process of new test methods for
Conference discussions. In addi-         hazard assessment.
tion, the Conference was addressed
by a number of high level officials,     Intensive discussions of animal wel-
including the State Secretary for        fare concerns has led to consensus
Agriculture, Food and Fisheries of       on issues such as the provisional,
Sweden, the Director-General of the      rather than definitive, acceptance of
Swedish      National   Chemicals        methods for human health assess-
Inspectorate, and the Deputy             ment in those cases where no
Assistant Administrator of the US        human reference data is available to
EPA.                                     confirm the relevance of the new
                                         method.        In addition, the
The agreements reached in                Conference agreed on practical
Stockholm include detailed guid-         guidance for the application of the

                                                                                                     NCA - Newsletter - 11
  Russell and Burch Award
The Humane Society of the United States (HSUS) pre-           the important role that scientists themselves can and do
sents the Russell and Burch Award to scientists who           play in advancing the cause of animal protection in
have made outstanding contributions toward the                laboratories. Candidates for the award are judged on the
advancement of alternative methods in the areas of bio-       scientific merit of their contribution to the alternatives
medical research, testing, or higher education.               field and on their professional commitment to this field.
Alternative methods are those that can replace or reduce      Applicants should have a history of laboratory work that
the use of animals in specific procedures, or refine pro-     is above reproach on humane grounds.
cedures so that animals experience less pain or suffer-       Send nominations by May 15, 2002 to: Russell & Burch
ing. The award, which carries a $5,000 prize, is named        Award, Animal Research Issues Section, The HSUS, 2100
in honor of William M. Russell and Rex L. Burch, the sci-     L street, NW, Washington, DC 20037, USA (tel.: 301-258-
entists who formulated the Three Rs approach of               3041, fax: 301-258-7760, e-mail: ari@hsus.org. No special
replacement, reduction, and refinement. Prior to 1999,        forms are necessary. Persons nominating themselves
The HSUS presented the award on an annual basis. It is        should submit a cover letter explaining their suitability
now bestowed every three years at the triennial World         for the award (see preceding paragraph), a curriculum
Congresses on Alternatives and Animal Use in the Life         vitae, and representative published articles. Persons
Sciences.                                                     nominating others should submit a letter explaining the
The next World Congress will be held August 2002 in           nominee’s suitability for the award and arrange to have
New Orleans, Louisiana. For more information, go to:          supporting documents forwarded. Winners are selected
http://www.worldcongress.net/                                 with the aid of an advisory panel.
The Russell and Burch Award is a means of recognizing




  NEW AALAS AWARD
  The Bennet J. Cohen Animal Stewardship Award
Dr. Ben Cohen, a founding father of AALAS, dedicated          Our professional community is richly endowed with
his professional life to improving the care offered to ani-   those who hold to the principles of proper and humane
mals used in teaching, testing in research. Dr. Cohen         animal care and use. I encourage you to identify some-
believed that good science and humane animal care not         one or some organization/institution who is deserving
only could co-exist, but must co-exist and that there is      of this distinguished honor and to submit a nomination
synergy between them. A leader within AALAS (includ-          on their behalf.
ing Past-President) and many other related professional       To nominate, one should submit a letter of nomination,
organizations, Ben passed away in 1991; yet, his many         additional supporting letters of nomination (no fewer
efforts and contributions form the basis today for            than four, nor more than ten) to the Awards Selection
humane animal care and use.                                   Committee, c/o the National AALAS office 9190
                                                              Crestwyn Hills Drive, Memphis, TN 38125-8538) no later
The AALAS Board of Trustees has voted to approve cre-         than May 1, 2002. Nomination letters should describe in
ation of the Bennet J. Cohen Animal Stewardship Award         detail the specific contributions and how these efforts
to recognize an individual or individuals, organization       have elevated the comfort and well being of animals or
or institution who have made an extraordinary effort          how the contributions have resulted in reduced num-
toward advancement of methods that replace or reduce          bers of animals used. If individuals are being nominat-
the number of animals used in research, teaching or           ed, the curriculum vitae of such individuals should also
testing, or for development of programs or procedures         be submitted at the same time as the nomination and
that elevate the comfort and well-being of such animals.      supporting letters.




NCA - Newsletter - 12
  Anny Eck-Hieff Prize
The “Fondation Internationale pour la Substitution de          Etats-Unis 35 L – 1477 LUXEMBOURG
l’Expérimentation Animale” (F.I.S.E.A.) founded in             The deadline for application is 31 May 2002.
Luxembourg on 13 December 1986 will award the                  The candidates have to introduce:
Annual European Prize for the Substitution of Animal           A summary of their research work (15 pages double-
experimentation “Anny Eck-Hieff Prize” in November             spaced, A4 format) in threefold. (A discussion is made
2002. The prize will be awarded to a researcher or a           of the value and the benefits of the proposed new alter-
team of researchers working on the key aspects of new          native method in relation to the experimental technique
alternative methods that eliminate or reduce the use of        using animals. The proposed technique has to be of real
animals or are less stressful to animals. The total fund-      and proven application); certificates, academic titles; a
ing of the Prize will be ¤ 7400 and can be divided in two      list of publications; and other relevant documents.
equal parts in the case there are different disciplines.
The prize is open to any researcher, medical doctor,           A board of international scientists and animal protec-
physicist, bio-mathematician, bio-engineer, etc.               tionists will designate the laureates. More information
Proposals have to be introduced to: F.I.S.E.A. Rue des         (in German) on: http://www.fisea.lu



  Meeting report - ECVAM/ESTIV TEACHING WORKSHOP ON VALIDATION

A workshop on validation, organ-         first day, finishing with a tour of the   last presentation, Manfred Liebsch
ised jointly by The European Centre      laboratory facilities at ECVAM.           discussed the regulatory acceptance
for the Validation of Alternative        Rodger Curren (Institute for In Vitro     of new alternative tests. Each pres-
Methods       (ECVAM)    and     the     Sciences, USA) introduced the con-        entation was followed by a thirty-
European Society of Toxicology In        cept of validation and explained          minute discussion period, and there
Vitro (ESTIV), was held at ECVAM         why it is necessary. This was fol-        was a general discussion at the end,
from 26 – 27 November 2001.              lowed by a talk given by Michael          during which several important
Some 20 participants attended the        Balls on selecting tests for valida-      issues emerged. One of the most
workshop from 12 different coun-         tion and organising validation stud-      important of these issues concerned
tries, with 8 speakers. The work-        ies. Andrew Worth (ECVAM) then            ways for improving communication
shop was organised as part of the        discussed the importance of predic-       between those involved in valida-
ongoing programme of validation          tion models. This presentation was        tion studies and scientists who are
workshops being organised by             followed by the first of two talks on     developing new test methods. Also,
ESTIV for its members, and also as       how to manage a validation study          it was agreed that there need to be
part of the ECVAM initiative for         and the lessons learnt so far, given      improvements in the way in which
PECO countries, those in the             by Horst Spielmann (ZEBET), with          general access to information on
process of joining the European          reference to the photo toxicity and       alternatives is available. It is the
Union (EU). The principal objective      embryo toxicity studies. The next         intention that the participants and
of the workshop was to act as a          day of the meeting started with the       the lecturers at the workshop will
forum for individuals from industry,     second of the talks on managing a         form a network to help with these
academia and regulatory bodies to        validation study, when Manfred            issues and to improve communica-
be brought up to date with the cur-      Liebsch (ZEBET) discussed the skin        tions and the provision of informa-
rent state of the art on validation,     corrosivity and skin irritation stud-     tion. The workshop is the first in a
and to hear international experts,       ies. This was then followed by two        series of similar meetings that are
some of whom have managed suc-           presentations on specialised topics.      being planned to take place over
cessful validation studies, present      The first of these concerned the          the next few years.
and debate this important issue.         validation of expert systems, given       Bob Combes
Michael Balls (ECVAM) introduced         by      Martin      Barratt    (Marlin    ESTIV
the first day, and Robert Combes         Consultancy, UK), and then Ludwig         FRAME
(ESTIV and FRAME) chaired the            Hothorn gave a critical account of        Russel &Burch House
second day. The workshop com-            ways to statistically analyse the         96-98 North Sherwood Street,
prised four presentations on the         results of validation studies. In the     Nottingham NG1 4EE - UK

                                                                                                        NCA - Newsletter - 13
  Book review: Pain management in animals
Paul Flecknell and Avril Waterman-      ketamine, nitrous oxide and others.      The different pharmacological
Pearson (Editors), W.B. Saunders,       Upon discussing all of these agents,     options are described as an integral
2000 (256 pages, ISBN 0702017671,       primary characteristics and side         part of a complete anesthesia proto-
€ 35,-)                                 effects of the class od drugs are        col, and are illustrated by a prob-
                                        covered next to focusing on the          lem-oriented approach in the differ-
Recently, the publishing house W.B.     specific possibilities for application   ent animal species [dog, cat, horse
Saunders presented the book Pain        of these different drugs in analgesic    and bovine]. Also, the specifically
management in Animals edited by         treatment of numerous animal             suited methods for local or regional
dr Paul Flecknell, certainly no         species. The primary animal species      anesthesia techniques are described
unknown name in the laboratory          covered in the book are the com-         and clearly illustrated with respect
animal world, and his colleague the     panion animals and the horse, but        to the application of local anesthet-
veterinary anesthesiologist dr Avril    in a more compact way, relevant          ics or other locally applied anal-
Waterman Pearson.                       information is provided on other         gesic such as opioids.
In the well-structured book the dif-    animal species.                          Clear tables support the reader in
ferent aspects of the problem of        Since a proper clinical evaluation of    finding not only the right [type of]
Pain in Animals are fully covered.      the analgesic effect is crucial for      drug, as such depending on the
In the first chapter, one of the edi-   determining the drug’s efficacy,         level of severity of the pain, but
tors [PF] outlines the relevant         chapter 4 eloquently covers many         also in choosing the right dose and
aspects of the discussion on the        aspects of pain behavior and pain        dosing interval in the different
topic of pain animals, carefully han-   assessment in animals [both qualita-     species. In conclusion, by reading
dling questions such as ‘Does an        tively as well as quantitatively].       and using this book, the reader is
animal feel pain?’ and if so, ‘When     On the one hand there are the            supported to not only differentiate
does this pain occur?’ and ‘How do      research data originating from anal-     between the different types of pain
animals demonstrate pain?’.             gesiometric studies, in which the        but also at the same time be able to
The next chapter focuses on the         stimulus is characterized precisely      design a well-balanced analgesic
pathophysiology of pain emphasiz-       thus allowing for an uniform             protocol for a specific pain problem
ing the different background            response in the animals tested. On       in a specific animal.
mechanisms and processes that           the other hand there is the descrip-     By choosing the right drug and
play a major role in the develop-       tion of clinical parameters used to      using a correct dose and dosing
ment of pain. Concepts and phe-         evaluate clinical pain, both acute       interval for the different species,
nomena such as [peripheral and          and chronic.                             achieving an adequate pain control
central] sensitization are covered in   Next to a more general description,      is greatly supported.
adequate detail. An in-depth under-     the different species-dependent          Despite the fact that this book pri-
standing of these phenomena is          characteristics of pain behavior are     marily aims for the veterinary prac-
essential in fully appreciating the     described, with a primary focus on       titioner, other professionals that are
necessity of an adequate but also       species such as dog, cat, and the        confronted with the problem of
timely analgesic therapy.               horse and to a lesser extent species     pain in animals can easily benefit
A comprehensive overview of the         such as ruminants, swine, rabbits        from the information in this book,
pharmacology of the different           and rodents. The authors of this         and thereby more adequately
painkillers is presented by dr.         chapter are to be commended for          design and implement pain control
Andrea Nolan [professor of pharma-      the effort they put into this chapter    strategies in the animal population
cology in Glasgow] in chapter 3.        by providing available information       in their care.
One by one, the different relevant      on non-mammalian species such as
pharmacological aspects of opiate-      birds, fish, amphibians and reptiles.    L.J. Hellebrekers
analgesics, the NSAID’s and the         In the following chapters the differ-    Professor of Veterinary
local analgesics are covered in-        ent aspects from the earlier chapters    Anesthesiology
depth. Next to the topics mentioned     are combined to construct a series       Utrecht University
above, there is a discussion on rele-   of practical pain control strategies     PO Box 80154
vant characteristics of alpha-2         aimed at both acute or post-opera-       3508 TD Utrecht
adrenergic agents and drugs such as     tive pain as well as chronic pain        The Netherlands

NCA - Newsletter - 14
  The European Resource Centre for Alternatives
  to animal use in higher education (EURCA)
The European Resource Centre for Alternatives to animal use in higher education
(EURCA) actively promotes the use of animal free models in education through outreach
activities (e.g. taking the Resource Centre to relevant scientific meetings) and through a
website (http://www.eurca.org) containing a content-rich database.


T    arget groups of the Centre are
     lecturers at institutes of higher
education, college and university
                                         2.
                                         3.
                                         4.
                                             Frog Heart
                                             Langendorff Heart
                                             Muscle Physiology
                                                                                   Additional educational resour-
                                                                                   ces – possibility to win an AWARD
                                                                                   EURCA is particularly interested in
students in the biomedical (pharma-      5.  Experiments in Human Neuro-           examples of teaching materials
cology, physiology, anatomy) and             Muscular Physiology                   which have been produced to sup-
veterinary sciences, and laboratory      6. Mouse       Watch      PVC   Rat       port the use of alternatives. These
technicians. Through its activities,         Simulations of Pharmacological        additional resources may be teach-
EURCA aims to support teachers to            Experiments on Rabbit Jejunum         ing notes that you have developed
be more creative in their approach       7. Simulations of Pharmacological         to support the use of specific alter-
to teaching and learning, to foster          Experiments on the Guinea Pig         native, workbooks, self-assessment
high quality training for science stu-       Ileum                                 questions, etc. If you have any spe-
dents and to significantly reduce the    8. Sniffy - the virtual rat               cific support material for one or
numbers of animals used unneces-         9. Vertebrate Dissection Guide -          more of the alternatives in our
sarily in teaching. An Advisory              the Dogfish                           database (or perhaps for alterna-
Board and a Dutch Project Team           10. Nerve Physiology                      tives that are not yet described in
reflect and advice on EURCA’s            Reviews will also soon be available       our database) and you would like
activities. The EURCA project is a       for:                                      to share these with other users,
result of the recommendations in         • SimHeart                                please submit them to EURCA
the ECVAM workshop report 33:            • Cellular respiration                    before 29 April 2002.
Alternatives to the use of animals in    • SimMuscle                               THE BEST IDEA FOR ADDITIONAL
higher education (ATLA 27:39-52,         • Anaesthesia of rats                     RESOURCES WILL WIN AN AWARD
1999).                                   • Vertebrate dissection guide - the rat   OF €100,-
                                         • The rat, a practical dissection guide   Criteria that will be sued to judge
Website                                  EURCA is actively requesting              the ideas include:
The EURCA website (http://www.           ‘experts’ in the field to review alter-   • The generic usefulness of the
eurca.org) offers extensive product      natives in the database and would             support materials in teaching
descriptions, features reviews, addi-    be keen to enrol additional review-       • The availability of the support
tional educational resources, addi-      ers to write these.                           materials to other users;
tional descriptions and user com-                                                  • The contribution the support
ments. The website also holds            Future website development:                   materials make to one of the
background information about ani-        discussion list                               three R’s
mals in education, an advisors list, a   One of the ways EURCA would like          • Any evaluative data which
discussion list (soon to be devel-       to make the website more interac-             demonstrate the effectiveness of
oped), links to other interesting        tive is to add a users discussion list        the teaching session
websites, sponsor information, col-      where comments and questions on           • Evidence that the use of the
laborating organisations and contact     educational alternatives could be             alternative(s) has(ve) reduced
details.                                 posted. Topics could include:                 animal use
                                         development of alternatives, experi-
Reviews                                  ences of using alternatives in teach-     Resource Centre outreach
The EURCA database currently             ing, technical issues, or specific        Representatives from EURCA (Dr.
holds reviews (most of which have        subject-related discussions               David Dewhurst, Dr. Jan van der
been specially commissioned) for:        Statements or questions may initiate      Valk or Jasmijn de Boo) attend con-
1. Blood Physiology                      the discussion.                           gresses and symposia. Presentations

                                                                                                        NCA - Newsletter - 15
are supported with demonstrations                    organised by ECVAM in light of             For more information on EURCA’s
of computer-based educational                        the PECO program (EU pre-                  activities, contact:
resources and 3D models.                             association countries).
                                                   • 8th FELASA Symposium 2002, 17-             Jasmijn de Boo
Events                                               20 June, Aachen, Germany                   EURCA
In 2002, EURCA will attend or par-                 • XIVth World Congress of Pharma-            NCA
ticipate in the following congresses                 cology IUPHAR 2002, 7-12 July,             Yalelaan 17
and symposia:                                        San Fransisco, CA, USA                     3584 CL Utrecht
• Workshop “Alternatives to the                    • Fourth World Congress on                   The Netherlands
    use of animals in higher educa-                  Alternatives and Animal Use in             E-mail: j.deboo@vet.uu.nl
    tion”, 5-6 april 2002, Piran,                    the Life Sciences, 11-15 August,
    Slovenia. This workshop will be                  New Orleans, Louisiana, USA



                                                     Newsflashes
New URL NCA website                                to present basic facts and principles that   Course coordinator, Department of
The website of the NCA has now a new               are essential both for the humane use        Laboratory Animal Science, Faculty of
identifiable URL:                                  and care of animals and for the quality      Veterinary Medicine, P.O. Box 80.155,
http://www.nca-nl.org                              of research.                                 3508 TD Utrecht, The Netherlands.
Both the new and the old URL will                  The course will consist of lectures,         Tel. +31 – 30 – 253 20 33.
direct you to the NCA web pages.                   hands-on practicals, workshops, and          Fax: +31 – 30 – 253 79 97.
                                                   demonstrations. It will cover a wide         E-mail: pdk@las.vet.uu.nl.
Anny-Heck-Hieff Award 2001                         range of topics, including recognition of
On November 10 2001, the 15 Anny-
                        th           th
                                                   pain and distress, anaesthesia and anal-     Rats mice and birds final rule delayed
Heck-Hieff Award (formerly the FISEA               gesia, euthanasia, humane endpoints,         another year in the USA
Award) was distributed in Luxembourg               husbandry, diseases, nutrition, trans-       A provision that prohibits the U.S.
by the Minister of Research, Erna                  genic animals, ethical aspects, and alter-   Department of Agriculture (USDA) from
Hennicot-Schoepges. The price was                  natives to animal experiments. A pre-        issuing a final rule on coverage of rats,
awarded to Birgid Neumeister and to                liminary program description is given        mice, and birds under the Animal
Maria Wartenburg and Heinrich Sauer.               below.                                       Welfare Act (AWA) regulations for
Birgid     Neumeister        (University     of    The course will run from May 27 – June       another year was included in the final
Tübingen,       Germany)      received      the    7, 2002. In addition, several optional       House-Senate conference report on the
award for her work to replace mice and             two-day modules, scheduled directly          agriculture appropriations bill for FY
guinea pigs in her work on legionella              after the course (June 10-11), will offer    2002.
infections with in vitro methods (Appl.            in-depth information on the following        The conference report language is as
and Env. Microbiology 66, 914, 2000).              topics: anaesthesiology, microsurgery,       follows: ‘None of the funds appropriat-
Maria Wartenburg and Heinrich Sauer                nutrition, genetic monitoring, and pri-      ed or made available by the Act shall be
(University of Köln, Germany) received             matology.                                    used to issue a proposed rule for which
the award for their work on tumor-                 The contents of the course are in line       the comment period would close prior
induced angiogenesis in confrontation              with     recommendations        of    the    to September 30, 2002, final, or interim
cultures of multicellular tumor sphe-              Federation of European Laboratory            final rule pursuant to notice and com-
roids and embryoid bodies grown from               Animal Science Associations (FELASA)         ment rulemaking in relation to any
pluripotent       embryonic       stem     cells   regarding the training of the young sci-     change or modification of the definition
(FASEB journal 15, 995, 2001).                     entist whose research involves the use       of ‘animal’ in existing regulations pur-
                                                   of vertebrate animals. The course may        suant to the Animal Welfare Act.”
Two-week        Course       to   Focus     on     also be useful for those interested in       Effectively, the language allows USDA
Humane Care and Use of Lab Animals                 setting up similar programs elsewhere.       to begin internal drafting of a proposed
The University of Utrecht in the                   Sessions will be conducted in English.       rule; however, no proposal could be
Netherlands is offering a two-week                 Applications are due by April 1, 2002.       finalized before October 1, 2002.
intensive course on laboratory animal              For information and application forms,
science. The objective of this course is           contact Stephan van Meulebrouck,

NCA - Newsletter - 16
  Development of an in vitro test battery for the estimation of
  acute human systemic toxicity; Outlines of the EDIT project

The aim of the multicenter programme EDIT (Evaluation-                            The results from the MEIC pro-
guided development of in vitro test batteries), is to estab-                      gramme indicated that the predic-
lish and validate new in vitro tests on kinetics and organ                        tion of human lethal concentrations
specific toxicity to be incorporated into optimal batteries                       increased considerably if human
for estimation of human acute systemic toxicity.                                  kinetic data (passage across the
                                                                                  blood-brain barrier and timing of
The scientific basis of the EDIT pro-    will be carried through by a com-        the lethal action) was used together
gramme is the good prediction            mon step-wise procedure: The             with the EC50-values. Therefore,
(R2=0.77) of human acute toxicity        Scientific committee of the EDIT         the establishment and validation of
by three 24-hour human cell line         programme will identify the need         computer-modelled simulations as
assays      (cell     type/endpoint:     for specific sets of toxicity or toxi-   well as in vitro models for key
HepG2/protein content, HL-60/ATP         cokinetic data. Laboratories are         kinetic events such as passage
content, and Chang liver cells/mor-      then invited to perform the defined      across the BBB, biotransformation,
phology and pH changes), found in        tests in order to provide the “miss-     and distribution are priority areas
the MEIC (Multicenter Evaluation of      ing” data for the 20 EDIT reference      within the EDIT kinetic programme.
In vitro Cytotoxicity tests) pro-        chemicals (a sub-set of the 50 MEIC
gramme. The MEIC study was initi-        reference chemicals). Expertrådet        Laboratories that are interested in
ated and directed by Björn Ekwall        and the EDIT Scientific Committee        participating in the EDIT pro-
and the aim was to evaluate the          will be responsible for proper           gramme are welcome to contact
relevance of in vitro cytotoxicity       evaluation of all submitted in vitro     Cecilia Clemedson for more infor-
tests for predicting human acute         results within the EDIT programme.       mation.
systemic toxicity.                       The submitted results will be evalu-
The results from the MEIC study          ated against the MEMO database           Cecilia Clemedson
also indicated that at least two types   (the MEIC Monograph programme),          Expertrådet AB
of in vitro tests ought to be added      i.e. the human acute systemic lethal     Högklintavägen 7
to the existing test battery in order    and toxicity data compiled within        172 64 Sundbyberg
to improve the prediction of human       the MEIC programme.                      Sweden
acute systemic toxicity. These tests                                              cecilia.clemedson@expertradet.se
should be able to determine key          EDIT models for target organ tox-
kinetic events, and crucial organ-       icity and kinetics
specific mechanisms not covered by       In the MEIC study the correlation
the tests in the MEIC battery. The       between in vitro EC50 values and
EDIT programme was initiated in          human lethal blood concentrations
order to develop and validate these      for acute toxicity was good for the
types of tests. Björn Ekwall was the     majority of the 50 reference chemi-
co-ordinator of the EDIT project         cals. However, for some chemicals
until his death in August 2000. The      the acute lethal blood concentra-
Scandinavian Society for Cell            tions in humans were under-pre-
Toxicology (SSCT) has recently           dicted by the in vitro methods. This
decided to continue the project. A       was explained by the target-interac-
Scientific Committee that will organ-    tion with specific receptors or
ise the study has been elected and       mechanisms of these chemicals.
the new co-ordinator of the project      Since several of the under-predicted
is Cecilia Clemedson at Expertrådet      chemicals have a neurotoxic poten-
AB, Sundbyberg, Sweden.                  tial the development within the
The EDIT programme will be a             EDIT target organ programme ini-
case-by-case study, but the estab-       tially should be focused on the
lishment and validation of new tests     nervous system.

                                                                                                       NCA - Newsletter - 17
   The Dutch Programme on Alternatives to
   Animal Testing “Dierproeven Begrensd”

T     he ZonMw1 Programme Committee
      on        Alternatives
Experiments has published its research
                                       to      Animal
                                                                  ment and reduction of animal experi-
                                                                  ments. It is awaited that the results of
                                                                  these projects will have a substantial
                                                                                                                               On behalf of sponsors of the pro-
                                                                                                                               gramme researchers are invited to sub-
                                                                                                                               mit projects dealing with ecotoxicology
programme on the 3R-research in the                               influence in the implementation of                           and     defence     related     research.
Netherlands. The programme is titled                              alternatives in the daily practice.                          Furthermore projects developing a
“Dierproeven begrensd”, which means                               The second area of consideration is                          monitorsystem, which identifies the
“animals experiments delimited”. The                              “Guideline Testing”. This area compris-                      relations between the implementation
programme is sponsored by the Dutch                               es animal experiments that are per-                          of alternative methods and the number
government, industry, health care foun-                           formed for regulatory purposes, legal                        of animal experiments are requested.
dations and animal welfare organisa-                              rules and obligations. Due to the inter-                     The last area of consideration is called
tions.                                                            national dimensions of the legislation                       “Dissemination”. Implementation of
The programme succeeds the sponsor-                               development and validation of alterna-                       alternative methods in the daily practice
ing      activities      of    the      Forum        for          tive methods might induce a consider-                        depends also on an adequate dissemi-
Alternatives to Animal Experiments,                               able reduction in animal experiments                         nation of project and programme
which supported the development of                                both in the Netherlands and abroad.                          results. It is the opinion of the pro-
alternative methods in the period 1987-                           Another area of consideration is the                         gramme committee that dissemination
1999. Although the new programme                                  area dealing with the development of                         should not only be limited to the scien-
supports the development and valida-                              “Research Models”.                                           tific community. Projects dealing with
tion of alternatives in general, much                             Approximately half of the animal exper-                      the broad dissemination of outcome of
more emphasis will be given to the                                iments in the Netherlands is carried out                     projects are welcomed.
implementation of alternative methods.                            in science projects. An important part of                    The programme committee will launch
Based on evaluations of funding policy                            this research induces moderate to                            three rounds of grant applications. For
and quick-scan inventories six areas of                           severe pain and stress in experimental                       each round approximately € 900.000
consideration have been identified.                               animals. It is also recognised that ani-                     will be available. In 2001 the pro-
Especially projects aiming the develop-                           mal models have limitations for the                          gramme      committee     received     43
ment and implementation of alterna-                               research to complex disorders. For that                      requests for funding. Based on the lim-
tives in these areas will be supported.                           reason there is a need for non-animal                        ited budget, scientific evaluation, social
Furthermore,            projects         developing               research methods, for alternative mod-                       relevance and the change of implemen-
“humane endpoints” will have special                              els to study human diseases and for                          tation, the committee granted 8 projects.
interest of the Programme Committee.                              models          using         human            tissues.      One project concerns education, anoth-
The first area of consideration is “sera                          Furthermore research on pain control                         er project deals with the development
and vaccines”. An important part of the                           and relief in experimental animals is                        of humane endpoints, a third project
animal experiments in the Netherlands                             needed as well as research on the                            concerns the development of alterna-
is performed for the development, pro-                            effects of application of new technolo-                      tives in the area of sera and vaccines,
duction and control research of sera,                             gies in experimental animals, e.g. func-                     and five other projects deal with the
vaccines and other biological products                            tional genomics, proteonomics and                            development of new research models.
for man and animals. A lot of these                               other biotech methods.                                       The next round will start in the first
experiments affect animal welfare in a                            The programme committee welcomes                             quarter of 2002.
severe manner. Animal experiments in                              also projects from the area “Education”.
this area are often very complex.                                 Although the absolute numbers of                             Information:
Although support will be given to pro-                            experimental animals used for educa-                         Dr. Arthur van Iersel
jects aiming on the replacement of                                tion is limited, it is believed that the                     Programme Committee on Alternatives
these animal experiments, priority will                           introduction and use of alternative                          to Animal Experiments
be given to projects that develop and                             methods will have a major impact on                          ZonMW
validate methods resulting in refine-                             the attitude of future researchers.                          Postbus 93245
                                                                                                                               2509 AE Den Haag
1 The organisations ZorgOnderzoek Nederland (Health Care and Development Council) and NWO-Medische Wetenschappen
(NWO- Medical Sciences) are associated in the new organisation ZonMw since June 2001. ZonMw conducts programmes cov-           The Netherlands
ering the broad area of health and diseases, prevention and health care. The principal sponsors of ZonMw are the Ministry of
Public Health, Welfare and Sport and NWO                                                                                       E-mail: iersel@zonmw.nl

NCA - Newsletter - 18
  OECD expert panel recommends replacing animal tests by
  cell and tissue tests for the evaluation of phototoxic and
  corrosive properties of chemicals (BgVV Press release)

From 30 October – 2 November            ommendation of the OECD expert           for phototoxic potential has to be
2001, two Nominated OECD                panel at their next national co-ordi-    conducted in particular for drugs
(Organisation for Economic Co-          nators meeting next spring, OECD         and UV-filter chemicals, which are
operation and Development) Expert       members states, the major produc-        used as sunblockers in cosmetic
Commissions have met at the BgVV        ers of industrial chemicals, will fol-   products.
(Federal Institute for Health           low the advice of the experts and        The two other TGs allow using tis-
Protection of Consumers and             officially accept the new cell and       sue tests to measure skin corrosion
Veterinary Medicine) in Berlin to       tissue tests to the OECD TGs for the     - the production of irreversible
evaluate two draft Test Guidelines      testing of chemicals. Acceptance of      damage in the skin following the
(TG) submitted by the European          the new non-animal tests by the          application of a test material. In the
Union to replace internationally        OECD is important, since results         human skin model test, a test mate-
accepted animal tests for the assess-   obtained by testing according to         rial is applied topically for up to
ment of hazardous chemicals by          OECD TGs are accepted world-             four hours to a three-dimensional
non-animal tests employing cells        wide by regulatory authorities. So       human skin model, which is grown
and tissues. The tests have to be       far, results obtained by the three       from human skin cells and can be
conducted for regulatory purposes       non-animal tests were only accept-       commercially obtained. In the sec-
to evaluate the phototoxic and cor-     ed by the regulatory authorities of      ond test, the rat skin transcutaneous
rosive potential of industrial chemi-   member states of the European            electrical resistance (TER) assay, the
cals. Already in the year 2000 the      Union. As a result of the consensus      test material is applied for up to 24
European Union has accepted non-        reached, animal tests will not be        hours to the epidermal surfaces of
animal cell and tissue tests, which     permitted to assess the phototoxici-     skin samples taken from the pelts of
had experimentally been evaluated       ty and skin corrosivity even for         humanely killed young rats. Both
in experienced laboratories in          chemicals, which are exported to         tests are able to discriminate reli-
Europe for their reproducibility and    countries outside the European           ably between known corrosive and
relevance in the hazard assessment      Union.                                   non-corrosive chemicals.
of chemicals (BgVV press release        Phototoxicity refers to a toxic          Corrosivity testing has to be con-
5/2000).                                response that occurs when the skin       ducted to allow the safe transporta-
Scientists of the OECD expert com-      is exposed to chemicals and then         tion of bulk chemicals by ship, air-
mittee from Europe, Korea, Japan        subsequently exposed to UV or visi-      plane, train or truck.
and the USA have for the first time     ble light. The newly approved 3T3        The approval of the new cell and
recommended to the OECD mem-            NRU phototoxicity test, allows to        tissue tests by the OECD expert
ber countries to replace the estab-     correctly predicting the phototoxic      panel represents a major step for-
lished animal tests by experimental-    potential of chemicals to the human      ward for those who have worked
ly validated non-animal test, since     skin. The new test was developed         long and hard towards the valida-
they provide the same level of          and experimentally validated under       tion and acceptance of new meth-
information for the assessment of       the leadership of ZEBET, the             ods. Scientists at the BgVV have in
toxic properties of hazardous           national German centre for docu-         particular contributed to the devel-
chemicals. This progress in replac-     mentation and evaluation of alter-       opment and validation of two of the
ing animal tests by non-animal          natives to testing in animals, at the    OECD approved methods, the in
methods proves that the protection      BgVV. The experimental validation        vitro phototoxicity test and the
of consumers and workers, who are       of the new test was conducted in a       human skin model based corrosivi-
exposed to chemicals, is not            joint study by ECVAM, the EU             ty test.
reduced, when stressful animal test     Centre for the Validation of
are replaced by validated cell and      Alternative Methods, and COLIPA,         More information may be obtained
tissue tests.                           the European Cosmetic, Toiletry          from the OECD at:
As a result of the unanimous rec-       and Perfumery Association. Testing       http://www.oecd.org/ehs/testguid

                                                                                                       NCA - Newsletter - 19
   De stichting Bouwstenen voor Dierenbescherming roept onderzoekers op mee te dingen naar de

     Hugo van Poelgeest-prijs Alternatieven voor dierproeven

Groot 10.000 Euro (waarvan ten         • De ontwikkelde alternatieve          ouder dan 4 jaar, waarin het alter-
minste 3.300 Euro te besteden in de      procedure                            natief wordt beschreven en/of
privé-sfeer) voor een aantoonbare      • De concrete vermindering in          wordt toegepast.
bijdrage aan de vermindering van         diergebruik en de vermindering       Na     beoordeling       door    een
proefdiergebruik (en/of vervanging       in het lijden van het individuele    onafhankelijke wetenschappelijke
door lagere dieren of vermindering       dier.                                jury zal de prijsuitreiking door dhr.
van proefdierleed in dierexperi-       • De energie die gestoken is in de     drs.     B.     Van       den    Bos
menteel onderzoek met behoud             ontwikkeling van dit alternatief     (Europarlementariër en voorzitter
van de kwaliteit van de door het         (duur, betrokkenen, bijdrage         van     de   Programmacommissie
alternatief verkregen gegevens.          van de instelling of andere sub-     Alternatieven voor Dierproeven van
De inzenders dienen verbonden te         sidiegevers)                         NWO/MW) plaatsvinden op 7 juni
zijn aan (of werken in opdracht        • De           validatie         en    2002 op kasteel “Oud Poelgeest”.
van) een instelling waar dier-           toepassing/acceptatie van het
proeven worden uitgevoerd.               alternatief in en wellicht buiten    Inzenden voor 15 april 2002         in
In de aanmelding dienen de vol-          de onderzoeksgroep.                  zesvoud:    Centrum       Dier       &
gende zaken toegelicht te worden:                                             Maatschappij, t.a.v. prof.dr. Tj.   de
• De oorspronkelijke dierproef         De aanmelding dient vergezeld te       Cock Buning, Yalelaan 17, 3584      CL
    (doel, methode, aantal dieren      gaan van publicaties (of geac-         Utrecht.
    per jaar)                          cepteerde manuscripten), niet




  NCA-Themamiddag
  Alternatievendeskundigen
Datum: 23 mei 2002                     jaar een bijeenkomst voor DEC-         In de afweging die door een DEC
Lokatie: Faculteit Diergeneeskunde     leden en andere geïnteresseerden,      dient te worden gemaakt, speelt de
Utrecht                                maar met name gericht op de alter-     beschikbaarheid aan mogelijke
Organisatie: Nationaal Centrum         natievendeskundigen.           Deze    alternatieven voor dierproeven een
Alternatieven voor dierproeven         bijeenkomsten zijn bedoeld om de       belangrijke rol. Hiervoor kan het
(NCA)                                  meest recente informatie met           noodzakelijk zijn informatie te
                                       betrekking tot alternatieven door te   verzamelen uit de literatuur of
Binnen de huidige DEC’s zijn ver-      geven, actuele problemen (case-        andere bronnen. Het raadplegen
schillende expertises vertegenwo-      studies) te bespreken en te streven    van bestaande databestanden met
ordigd. Een van deze expertises is     naar een uniforme werkwijze            behulp van gerichte zoekwoorden
“alternatieven voor dierproeven”.      tussen DEC’s.                          is hierbij van groot belang. De
Het terrein alternatieven voor dier-    Naast de bespreking van een spec-     effectiviteit van deze zoekactie
proeven omvat een groot aantal         ifiek onderwerp, zal er op deze        wordt in belangrijke mate bepaald
deelgebieden met eigen specifieke      middag ruimte zijn om specifieke       door de omschrijving van de vraag-
problemen en ontwikkelingen.           onderwerpen of problemen in te         stelling. De themamiddag is er op
DEC-leden dienen een goede             brengen. Op 23 mei 2002 zal de         gericht informatie te verstrekken
afweging te kunnen maken bij de        tweede      bijeenkomst     worden     over de juiste zoekstrategieën in
beoordeling van aangemelde dier-       gehouden met als onderwerp:            diverse bronnen. Verder zullen
proeven. Het is daarom van belang                                             inleidingen worden gegeven over
optimaal geïnformeerd te zijn.         Zoekstrategieën in het kader van       specifieke databestanden met
Het NCA organiseert eenmaal per             literatuuronderzoek               betrekking tot alternatieven voor

NCA - Newsletter - 20
dierproeven. Tenslotte zal de          Tel. 030 2532186                     14.00 Ing.T.Bakker
mogelijkheid worden geboden om         e-mail: valk@las.vet.uu.nl                 (Hoofdafdeling
onder deskundige begeleiding een       Het inschrijvingsformulier voor            Proefdierkunde, UU):
en ander in de praktijk te brengen.    deze dag is te vinden op de NCA            Ontwikkelingen in zoek-
Vanwege de opzet van de middag is      website:                                   strategieën (onder voorbe-
het aantal plaatsen beperkt.           http://prex.las.vet.uu.nl/nca/             houd)
                                                                            14.30 Prof.dr.A.Goldberg (CAAT,
Informatie:                            Voorlopig programma:                       Johns Hopkins University,
Dr.J.v.d.Valk                          13.00 Ontvangst en koffie                  USA): ALT-web
Nationaal Centrum Alternatieven        13.30 Inleiding                      14.55 Koffie
voor        dierproeven  (NCA)         13.40 Drs.W.deLeeuw                  15.25 Dr.B.Grune-Wolff (ZEBET,
Universiteit Utrecht                         (Keuringsdienst van Waren):          BgVV, D): ZEBET database
Yalelaan 17                                  Actuele informatie van de      15.50 Practische oefeningen
3584 CL Utrecht                              KvW                            17.00 Einde




                                  Forthcoming events
Scand-LAS 2002. Uniting science        Research, Animals and Welfare A      Fourth world congress on alter-
and the three R’s                      training course for Ph.D. stu-       natives and animal use in the life
   18-21 April 2002                    dents from the Baltic, the Nordic    sciences
   Gardermoen, Norway                  countries, and North West                4-8 August 2002
   Information:                        Russia                                   New Orleans, USA
   http://oslovet.veths.no/scand-         May 27-June 5 2002                    Information: Donna Pease:
   las/2002/                              St. Petersburg, Russia                dpease@hsus.org, url:
                                          Information:                          http://www.worldcongress.net/
Ethics of Research with Human             http://www.uku.fi/~tnevalai/St.
Subjects                                  Pete/                             EUROTOX 2002 - the 40th
   May - July 2002                     Symposium: Doden van dieren          Congress of the European soci-
   Internet-based Seminar                 18 Juni 2002                      eties of toxicology
   Information: http://poynter.indi-      Garderen                              15-18 September 2002
   ana.edu/sas/                           Informatie:                           Budapest, Hungary.
                                          http://www.vet.uu.nl/congres/d        Information: http://www.euro-
Themamiddag Alternatieven                 odendieren                            tox2002.org/
deskundigen: Zoekstrategieën in                                             INVITOX 2002
het kader van literatuuronder-         Laboratory animal science - Basis        16-19 October 2002
zoek                                   and strategy for animal experi-          Formia, Italy
   23 May 2002                         mentation                                Information:
   Utrecht, The Netherlands            17-20 June 2002                          http://www.xs4all.nl/~shorbach
   Informatie:                            Aachen, Germany                       /INVITOX
   http://prex.las.vet.uu.nl/nca          Information: http://www.gv-
                                          solas.de/vera/ver1.html           FRAME reduction Committee sym-
Course on laboratory animal sci-                                            posium on reduction: current
ence                                                                        status and future prospects
   27 May-7 June, 2002                                                         16-17 October 2002
   Utrecht, The Netherlands                                                    London, UK
   Information: Mr. Stephen            2002 congress on in vitro biology       Information:
   Meulebrouck: pdk@las.vet.uu.nl         26-29 June 2002                      sylvia@frame.org.uk
                                          Coronado Springs, FL, USA
ECVAM status seminar 2002                 Information:                      2003 Congress on in vitro biology
   4-6 June 2001                          http://www.sivb.org/meeting.asp      31 May-5 June 2003
   Ispra, Italy                                                                Portland, OR, USA
   Information: Marlies Halder:        14th International congress of          Information:
   marlies.halder@jrc.it:              pharmacology                            http://www.sivb.org
                                          13-19 July 2002
                                          San Francisco, USA.               10th International congress of
                                          Information:                      toxicology
                                          http://www.iuphar2002.org/           11-16 July 2004
                                                                               Tampere, Finland
                                                                               Information: Ms Hanna Tähti:
                                                                               blhata@uta.fi , url:
                                                                               http://www.uta.fi/fst
                                                                                                NCA - Newsletter - 21
                          Colophon
                        NCA-Newsletter
                        (ISSN: 1387-1706)




                        Published by:
                        Netherlands Centre Alternatives to
                        Animal Use (NCA)
                        Dept. Animals & Society
                        Fac. Veterinary Sciences,
                        Utrecht University
                        Yalelaan 17
                        NL-3584 CL Utrecht
                        The Netherlands
                        Phone: +31 30 253 2186
                        Fax: +31 30 253 9227
                        E-mail: nca@pobox.uu.nl

                        Editors :
                        J. van der Valk
                        C. Hendriksen
                        I. Bos

                        Layout:
                        Jacqueline Zandvliet
                        JAB Design, Bilthoven

                        Printed by:
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                        Views and opinions from guest
                        authors may be different from
                        those of the editors.
                        For free subscription, contact the
                        editors.

                        Permission to reprint, please credit
                        author and NCA-Newsletter

                        The NCA-Newsletter is also avai-
                        lable at the website of the NCA:
                        http://prex.las.vet.uu.nl/nca




NCA - Newsletter - 22

				
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