1. Study consisted of 139 boys with hypospadias & 267 controls. We found no significant difference in weight (13.6±6.1 vs 14.8±6.8 kg, p=0.080), height (94.1±20.1 vs 93.5±19.9 cm, p=0.776) and age (45.12±35.53 vs 46.79±33.05 months, p=0.637) ASD was found to be significantly shorter in boys with hypospadias (40.00±9.68 vs 45.60±9.35, p<0.001). No significant difference was observed in AGD1 (78.91±12.33 cm vs 80.2±14.10 cm, p=0.367) and AGD 2 (69.90±13.06 cm vs 71.64±13.81 cm, p=0.273) (Fig 2) Age, weight and length adjusted regression model showed significantly shorter ASD (p <0.001) but not AGD1 (p=0.509) or AGD2 (p=0.149) in boys with hypospadias. We saw a negative correlation (r= -0.297, -0.267, -0.327, p<0.001) of severity of hypospadias with all three measures of AGD. The incidence of hypospadias has doubled over the past three decades. While exact etiology remains unknown, it appears to be a combination of genetic susceptibility and prenatal exposure to endocrine disruptors. Anogenital distance (AGD) is classically used to assign gender to animals and has been identified as one of the end-points in US Environmental Protection Agency guidelines for reproductive toxicity. Introduction Hypothesis Results Conclusions AGD is significantly shorter in boys with hypospadias and decreases with severity of hypospadias. This study strengthens the utility of AGD as a non-invasive predictor of male reproductive health outcomes. References Welsh, M., Saunders, P. T., Fisken, M. et al.: Identification in rats of a programming window for reproductive tract masculinization, disruption of which leads to hypospadias and cryptorchidism. J Clin Invest, 118: 1479, 2008 Toppari J, Virtanen HE, Main KM, Skakkebaek NE. Cryptorchidism and hypospadias as a sign of testicular dysgenesis syndrome (TDS): environmental connection. Birth Defects Res A Clin Mol Teratol 2010; Jain, V. G., Singal, A. K.: Shorter anogenital distance correlates with undescended testis: a detailed genital anthropometric analysis in human newborns. Hum Reprod, 28: 2343, 2013 Thankamony, A., Ong, K. K., Dunger, D. B. et al.: Anogenital distance from birth to 2 years: a population study. Environ Health Perspect, 117: 1786, 2009 88:910–919. Animal model of androgen disruption during early gestation shows correlation of subnormal AGD with hypospadias. Do boys with hypospadias have shorter AGD and does it correlate with severity? Animal studies have reported a critical time period, termed the ‘male programming window’ (MPW), during which genital development is programmed, likely to be 8–14 weeks of gestation in humans Once AGD gets programmed in the MPW by androgen action, it gets “fixed” and then grows in proportion to body growth independent of androgen levels Disruption of androgen action during this MPW has been reported to result in hypospadias. Subnormal AGD correlates with severity of hypospadias AGD may be a useful marker of androgen action during MPW and can provide life long readout of prenatal hormonal exposure Materials and Methods Prospective descriptive study involving all boys presenting with hypospadias The control group of healthy boys with normal genitalia was randomly selected from the adjoining well child check clinic. We measured AGD from the midpoint of anus to (1) ASD - bottom of scrotum, (2) AGD1 –penopubic junction (3) AGD2 – penoscrotal junction; in all boys. (Fig 1) Exclusion Criteria: Boys with buried penis, chordee without hypospadias, cryptorchidism, imperforate anus or any know syndrome were excluded from the study.