What happens when your hypothesis is…wrong? Listen as host Dr. Dan Tyrrell (University of Michigan Medical School) interviews lead author Dr. Julie Freed (Medical College of Wisconsin) and content expert Dr. Chi Fung Lee (Oklahoma Medical Research Foundation) about the new Rapid Report by SenthilKumar et al., which investigated the function of estradiol on human microvessels. In contrast to their hypothesis, Freed and co-authors found that exogenous estradiol treatment of arterioles isolated from both young and older women promoted endothelial dysfunction. In addition, the authors found that estradiol treatment of microvessels isolated from men led to endothelial and smooth muscle dysfunction. The timing of this article is key, given the new AJP-Heart and Circ requirements launched in January 2023 for considering sex as a biological variable. Freed and collaborators originally hypothesized that estrogen may activate the enzyme sphingosine kinase, and therefore mediate cardioprotective effects in the vasculature. However, that was not the case. Freed summed up their surprising results: “There is something going on here. Do we have all the answers yet? No. But we’re going to figure this out.” Listen as we discuss the complexities of human tissue banking, and the grit and flexibility necessary for all researchers to follow the science where it leads.   Gopika SenthilKumar, Boran Katunaric, Henry Bordas-Murphy, Micaela Young, Erin L. Doren, Mary E. Schulz, Michael E. Widlansky, and Julie K. Freed 17β-Estradiol Promotes Sex-Specific Dysfunction in Isolated Human Arterioles Am J Physiol Heart Circ Physiol, published January 6, 2023. DOI: 10.1152/ajpheart.00708.2022