Fifty seven normotensive male Sprague-Dawley rats (300 400 g), purchased from Harlan (Horst, The Netherlands), were maintained at a 12/12-h 12/12-hark cycle (with light beginning at 7 a.m).
Yeast strain R. toruloides (CBS 5490, Central Bureau voor Schimmelcultures, Utrecht, The Netherlands) were maintained on agar plates containing (per liter) 20 g of agar (Sigma, St . LouisMO, USA), 10 g of yeast extract (Biobasic Canada Inc ., 20 g of peptone (Biobasic Canada Inc). and 20 g of d-dextrose (Sigma, St . LouisMO, USA).
Eight to ten week-old C57Bl/6 male mice (Charles River, Maastricht, The Netherlands) were maintained at the animal care facility of the Academic Medical Centre, Amsterdam, The Netherlands according to institutional guidelines.
Eight-week-old male athymic nu/nu mice (Harlan, the Netherlands) were maintained under controlled pathogen-free environmental conditions 20-21°C 20-21°C% relative humidity and a 12-hour lighting cycle).
Young (1 to 3 months old) and old (20 to 24 months) male WKY rats (Harlan Netherlands) were maintained on a 12-h light-dark cycle and fed standard chow diet and tap water according to the local guidelines of animal experimentation.
AKN-1 (a gift from Dr A Nussler, University of Ulm, Germany), CC-LP-1 and CC-SW-1 (a gift from Dr P Bosma, AMC Liver Centre, University of Amsterdam, Netherlands) were maintained in DMEM with 10% FCS, L-glutamine (2 mM), penicillin (100 U/ml) and streptomycin (100 µg/ml) on uncoated flasks.
The HPV16 (SiHa), HPV18 (HeLa) positive cervical carcinoma and the mdm2 positive (SA-1) cell lines (from Institute of Human Genetics, Academic Medical Centre, Amsterdam, The Netherlands) were maintained in Dulbecco's Modified Eagle Medium (DMEM; Sigma, USA) supplemented with 10% fetal bovine serum (Sigma, USA) and used as controls.
The Netherlands were struggling in the conditions.
The Netherlands were partially metricated when they were French, in the years 1810 1813.
The Netherlands were populated by different groups.
