1. Estrogenic agents in pubertal female monkeys Mari Golub msgolub@ucdavis.edu

2. Systems that mature during in adolescence under the influence of estrogen Reproductive tract – Tissue growth, cell differentiation – Ovarian cyclicity, sex-differentiated morphometry Bone – growth and epiphyseal closure – mineralization Immune system – thymic involution – reproductive tract immunity – sex-differentiated immune function Brain – “formal operations” – risk taking – pruning of synapses, myelination of cortical connection – altered sleep patterns

3. Dosing for endocrine disruption study 12 months – 6 months before estimated age at menarche – 6 months after estimated age at menarche Comparison groups – Vehicle control – 25 mg/kg methoxychlor (MXC) – 50 mg/kg methoxychlor (MXC) – 0.5 mg/kg DES Route; feeding syringe, baby food vehicle

4. Estrogenicity of Treatments as determined in serum with in vitro ER  reporter assay 0 20 40 60 80 100 120 140 160 180 p g / m L DES MXC50 MXC25 CONTROL 0 20 40 60 total exogenous endogenous Estrogenicity 2 1224

5. Body weights Kilograms

6. Height increase during treatment 0 20 40 60 80 100 120 DES MXC50MXC25CONTROL * * Height (cm) Height (=crown rump + femur +tibia)

7. Number of Menstrual Periods 0 20 40 60 80 100 120 MXC50 MXC25 DES CONTROL Cumulative # of Periods Treatment Months

8. Nipple size 0 25 50 75 100 125 150 175 200 225 DES MXC50 MXC25 CONTROL 0 50 100 150 200 250 300 * * DES MXC50 MXC25 CONTROL * * -8 months after treatment - during treatment start treatment end treatment Volume (m 3 ) menarche

9. RBC hemoglobin content

10. Long term effects on leukocyte populations THS END DOSE CD56+/3- Time From Start of Dosing at 24 Months of Age Mean±SEM are shown *p<.03 * * n u m b e r o f c e l l s n u m b e r o f c e l l s

11. Ovarian cycles Ovarian ultrasound ControlMXC25MXC50DES Enlarged ovaries Ovarian mass/cyst 1 4 1 3 1 1 1 2

12. Conclusions Exogenous estrogen can influence growth and sexual maturation of adolescents It is difficult to characterize the effects on as a delay or acceleration of puberty Some effects persist six months after discontinuation of treatment These changes could be relevant for women’s reproductive function, immune function and disease risk

13. Collaborators Eric Gershwin Andy Hendrickx Carl Keen Lucille Hurley Bo Lonnerdal Dennis Styne Chuck Fuller Bill Lasley Chris Jerome SRAs Wil Saito Peter Takeuchi Laurie Brignolo Margaret Kaekuahiwi Hedi Arbabzedeh Stacey Germann Casey Hogrefe

14. Uterine ultrasound Ovarian ultrasound ControlMXC25MXC50DES Enlarged ovaries Ovarian mass/cyst 1 4 1 3 1 1 1 2 CRPRC: Uterus in anovulatory; Uterus /bw CRPRC: Uterus in anovulatory; Uterus /bw