As a secondary aim, we investigated whether

obesity parameters and the liver were affected by fructose feeding alone, using water-fed rats selleckchem as a control group. Bisphenol A (BPA), (80-05-7, (CH3)2C(C6H4OH)2, ≥99% purity), fructose (C6H12O6, ≥99% purity), Griess modified reagent, ZnSO4, and VCl3 were purchased from Sigma–Aldrich, St. Louis, MO. NaNO3 was purchased from Merck chemicals, Darmstadt, Germany. The animal study was approved by the Uppsala Animal Ethical Committee and followed the guidelines laid down by the Swedish Legislation on Animal Experimentation (Animal Welfare Act SFS1998:56) and European Union Legislation (Convention ETS123 and Directive 86/609/EEC). Sixty female F 344 rats at 3 weeks of age were purchased from Charles River International, Salzfeld, Germany, and housed 3 rats/cage at Uppsala University Hospital animal facility in a temperature-controlled and humidity-controlled room with a 12-h light/dark cycle. To minimize background BPA exposure Polysulfone IV cages (Eurostandard IV) and glass water bottles were used. The rats were fed a standard pellet RM1 diet (ad lib.) from NOVA-SCB, Sollentuna, Sweden. RM1 is a natural ingredient diet with a low level of phytoestrogens (100–200 μg/g)

(Jensen and Ritskes-Hoitinga, 2007 and Odum et al., 2001). During the two-week acclimatization period preceding the ten-week intervention all animals were given water to drink and during the intervention water or 5% fructose solution (see Section 2.3). At 5 weeks of age the rats were assigned to five groups (12 rats/group); water control (W), fructose control PD98059 in vivo (F), low dose BPA (0.025 mg/L), medium dose BPA (0.25 mg/L) or high dose BPA (2.5 mg/L). To avoid unnecessary stress no cage-mates were separated, but the cages were allocated to the different groups to achieve equality in weights in all groups. Food and liquid consumption in each cage and individual weight of the rats were determined once a week. Before

MRI exam, the rats were anesthetized with Ketalar 90 mg/kg bw (Pfizer, New York, NY) and Rompun 10 mg/kg bw (Bayer, Leverkusen, Germany). Immediately after the scanning they were killed by exsanguinations from the abdominal aorta while still under anesthesia. To prepare BPA exposure solutions (0.025, 0.25 and 2.5 mg/L), three stock solutions of BPA in 1% ethanol Isotretinoin (2.5 mg/L, 25 mg/L and 250 mg/L) were diluted 1:100 in 5% fructose solution. The low dose was chosen to be well below the recommended TDI, the medium dose corresponding to TDI (50 μg/kg and day), while the highest dose was ten times this level. The BPA was analyzed by liquid chromatography–tandem mass spectrometry by the Division of Occupational and Environmental Medicine in Lund, Sweden. The division is a European reference laboratory in the DEMOCOPHES EU project (www.eu-hbm.info/democophes) for analysis of BPA. The BPA concentrations in analyzed samples of the solutions were: water control – 0.