Between 1994 and 1995, 1 day nocturnal infusion of 5-fluorouracil (5-FU) was performed prior to surgery in 13 primary breast cancer patients; 300 mg/m2 of 5-FU was infused constantly from 2100 h to 0700 h via peripheral vein with a volumetric pump. 5-FU concentration in tissues was measured within surgical specimens by HPLC. The concentrations of 5-FU in tumor tissues ranged from 6 to 49 ng/g (average +/- SEM 25.0 +/- 4.1 ng/g), while in normal breast tissues and adipose tissues 5-FU was below the detection limit (<3 ng/g). The 5-FU concentration was lower in estrogen-receptor-positive tumors (14.4 +/- 4.5 ng/g) than in estrogen-receptor-negative tumors (31.8 +/- 5.0 ng/g). Typical FAA was observed in the tumor tissues of three patients. In these three cases, AgNORs were aggregated to one large spheroidal figure in more than 39% of tumor cells. Appearance of FAA could not be predicted by other clinical features. Nocturnal 5-FU infusion caused FAA changes in certain types of primary breast cancer.