A 4-nitrophenol (PNP) isolated from diesel exhaust particles (DEP) has been identified as a vasodilator. PNP is also a known degradation product of the insecticide parathion. We used uterotrophic and Hershberger assays to study the estrogenic and anti-androgenic activities of PNP in-vivo. In ovariectomized immature female rats injected subcutaneously with 1, 10, or 100 mg/kg PNP daily for 7 days, significant (P<0.05) increases in uterine weight were seen in only those receiving 10 or 100 mg/kg PNP. Furthermore, in castrated immature male rats implanted with a silastic tube (length, 5 mm) containing crystalline testosterone and injected subcutaneously with 0.01, 0.1, or 1 mg/kg PNP daily for 5 days, those receiving the doses of 0.1 mg/kg showed significant (P<0.05) weight decreases in seminal vesicles, ventral prostate, levator ani plus bulbocavernosus muscles, and glans penis. Plasma FSH and LH levels did not change in female rats but were significantly (P<0.05) increased in male rats treated with 0.1 mg/kg PNP. These results clearly demonstrated that PNP has estrogenic and anti-androgenic activities in-vivo. Our results therefore suggest that diesel exhaust emissions and the degradation of parathion can lead to accumulation of PNP in air, water, and soil and thus could have serious deleterious effects on wildlife and human health.