A comparison between humans and rabbits was performed based on stimulation of whole blood with well-known pyrogens from Gram-negative and Gram-positive bacteria, such as lipopolysaccharide (LPS) and lipoteichoic acid (LTA), respectively. The reactivity was measured as release of IL-1ß and IL-8 by ELISA. The reactivity of the two species towards LPS was found to be similar, whereas their reactivity towards LTA differed considerably. Differences between the levels of IL-1ß and IL-8 release were observed in both species. This finding suggests that the In vitro Pyrogen Test (IPT) which uses human blood to detect contaminations, e.g. of injectable drugs, might predict the human reaction to the contamination better than the "gold standard" rabbit pyrogen test.

Body temperature is a clinical parameter in vaccine quality control to detect systemic side-effects or to monitor progression of infectious diseases. Moreover, changes in body temperature are used as clinical parameters to define humane endpoints in animal experiments. However, measuring body temperature via the rectal route can be troublesome and distressing to the animal. Non-invasive measurement methods were developed in recent years. The aim of this investigation was to study and to compare rectally measured body temperature with data obtained with implanted temperature-sensitive transponders (TST) in mice, guinea pigs, rabbits and pigs under the controlled conditions of regulatory testing.

The endothelial cell layer is a multifunctional barrier between the blood stream and the vascular wall. Reactive oxygen species can damage endothelial cells and may so potentiate atherosclerotic lesion formation. Therefore, we established an in vitro model for the qualitative and quantitative investigation of oxidative DNA damage and repair in human coronary arterial endothelial cells (CAEC). Oxidative DNA damage was induced by standardised treatment with 50-400 µM hydrogen peroxide (H2O2). The amount of DNA damage was quantified by determination of DNA single strand breaks (SSB) and alkali-labile sites in individual cells, using the alkaline single-cell gel electrophoresis assay - "comet assay". Significant DNA damage could be induced reproducibly in CAEC cells after exposure to 50 µM H2O2 for 10 min. Additional treatment with catalase prevented DNA damage by H2O2. The time needed for DNA repair depended on the initial grade of damage. After 30 min post-incubation at 37°C, DNA damage was completely repaired in cells treated with 50 µM H2O2. Cell samples initially damaged with H2O2 concentrations between 100 µM and 400 µM were repaired after 60 min. This endothelial cell culture model allows experiments on oxidative DNA damage, alteration, and repair directly on the relevant target cells. Animals are used neither as direct objects of such experiments nor as cell or tissue donors.

Genotoxicity tests available today have several shortcomings. The widely applied Ames assay measures mutations in bacteria, thereby disregarding the physiological particularities of the human cell and organism.
We provide first evidence for a new concept of genotoxicity detection in living human cell cultures. The data were obtained by use of a newly developed assay, which is based on the quantification of fluorescent signals, i.e. counting of the relative number of fluorescent cells in the sample. It is characterised by a short reaction time and fulfills the requirements for automated performance. The new system monitors chromosomal rearrangements and, therefore, is predicted to detect a broad spectrum of genotoxic substances. Indeed, we demonstrate the genotoxic effect of ionising radiation, of an Ames assay positive compound, and of two compounds which are poorly mutagenic in the Ames assay. The new assay will be optimised further and adapted to the requirements for routine analysis in order to help to further reduce animal experimentation in genotoxicity testing.

The main objective of our institution is to establish a virtual laboratory on the Internet to allow for a reliable in silico estimation of harmful effects triggered by drugs, chemicals and their metabolites. In the past two years, we have compiled a pilot system including the 3D models of five receptors known to mediate adverse effects (the Ah, 5HT2A, cannabinoid, GABAA, and estrogen receptor, respectively) and tested them against 280 compounds (drugs, chemicals, toxins). Within this set-up we could demonstrate that our concept is able to both recognise toxic compounds substantially different from those used in the training set as well as to classify harmless compounds clearly as being non-toxic at low-level doses. This suggests that our approach can be used for the prediction of adverse effects of drug molecules and chemicals. It is the aim to provide free access to this 3D data base, particularly to universities, hospitals and regulatory bodies as it bears a significant potential to recognise hazardous compounds early in the development process and withdraw them from the evaluation pipeline. Hence, for substances recognised as hazardous in silico, subsequent toxicity tests involving animal models become obsolete.

NEWS
EU: Richtlinie 2003/15/EWG zur Änderung der Kosmetik-Richtlinie verabschiedet
EU: Dritter Report der EU zu Versuchstierzahlen liegt vor
CH: FFVFF vergibt zwei ALTEX-Preise 2002: Nina Schweigert et al. und Heinrich W. Grosse
EU/USA: InterNICHE vergibt einen "Humane Education Award 2003" für Indien
D: Professur für Ersatz- und Ergänzungsmethoden an der TiHo Hannover
D: DFG bewilligt Graduiertenkolleg Bioethik in Tübingen
D: RNA-Interferenz - gezieltes Abschalten von Genen in Zellkulturen?
D: Biochips für funktionelle Proteomik und Biomonitoring
D: BMBF fördert Verträglichkeitsstudie ohne Tierversuche
EU/USA: From guinea pig to computer mouse - Neuauflage
NL: Rückgang der Tierversuchszahlen 2001
USA: Alkohol erhöht Testosteronspiegel bei Ratten
UK: Parkinson-Forschung mit intelligenten Robotern
USA: ELAD® statt Xenotransplantation?
D: Stiftungsprofessur für Alternativen zum Tierversuch

CORNERS
MEGAT
ECVAM

CONFERENCE REPORT
London: Predictive toxicology: In silico modelling and expert systems
Bad Boll: Tierschutz in den Medien - Skandale, Tierschutz, Verantwortung
Edinburgh: Science in the service of animal welfare - UFAW Symposium 2003
Utrecht: Towards better in vitro methods: the replacement of fetal bovine serum

BOOK REVIEWS
Toni Lindl: Zell- und Gewebekultur (5. Auflage)
InterNICHE: From guinea pig to computer mouse (second edition)

COMMENTS
Martina Kuhtz, Brigitte Jenner, Gerald Hübner und Dieter Bütefisch: Weil wir keine
Ratten sind ...
Ursula G. Sauer, Horst Spielmann und Brigitte Rusche: Dritter EU-Bericht über die Anzahl verwendeter Versuchstiere - Tendenzen, Probleme, Schlussfolgerungen
Rainer Nobiling: Zur deutschen Versuchstierstatistik für biomedizinische Forschung

CALENDAR