In 1920, the male hormone turned boys into men, and the female hormone made women out of the girls. Feminists had won a major political victory in gaining the right to vote, and America had rid her shores of many foreign radicals. But out of this apparent calm, a new unrest soon broke loose. While feminism struggled to maintain its newfound identity, women’s roles contin­ued to change and sex hormones started to multiply.42

Three interrelated scientific questions took center-stage in the new re­search centers established in the 1920s. Which cells in the ovary or testis produced the substance or substances responsible for the sorts of effects Steinach, Moore, and others had observed? How could one chemically extract active hormones from these tissues? And finally, once one produced an active extract, could it be purified? In 1923, the biologists Edgar Allen and Edward A. Doisy, working at the Washington University Medical School in Saint Louis, announced the localization, extraction, and partial purification of an ovarian hormone.43 Just six years earlier Charles Stockard and George Papani­colaou (for whom the Pap smear is named) had developed an easy method to monitor the estrus cycle of the rodent.44 Allen and Doisy now used the tech­nique to assess the potency of extracts obtained from ovarian follicle fluid removed from hog ovaries.45 By injecting their extracts into spayed animals, they could try to induce changes in vaginal cells typical of rodents in estrus.

Purifying

figure 7.3 : Pregnant women’s urine has high concentrations of female hormone. (Source: Alyce Santoro, for the author)

First they showed that only substances from the fluid surrounding the oocyte in the ovary (called the follicular fluid) affected the estrus cycle. Not only did the spayed animals exhibit a change on the cellular level; they also changed behaviorally. Allen and Doisy noted that the animals displayed ‘‘typical mating instincts, the spayed females taking the initiative in courtship.’’ Having estab­lished a reliable method to test for hormone activity—called a bioassay, be­cause the test relies on the measurable response of a living organism—Allen and Doisy also tested extracts marketed by pharmaceutical companies. These turned out to have no bio-activity, justifying what they called a ‘‘well-founded skepticism concerning commercial preparations.’’46

Allen and Doisy had made a great start. They had a reliable bioassay. They had shown that the ovarian factor came from the liquid that filled the ovarian follicles (rather than, for example, the corpus luteum—another visible struc­ture in the ovary). But purification was another story. Progress was slow at first because the raw material was available in only limited quantities and at ‘‘staggering’’ costs. About 1,000 hog ovaries yielded 100 cubic centimeters (about a fifth of a pint) of follicular fluid, at the cost of approximately $1.00 per milligram of hormone.47 Then, in 1927, two German gynecologists dis­covered that urine from pregnant women has extremely high concentrations of the female hormone,48 and the race was on, first to gain access to enough of that suddenly valuable commodity (figure 7.3) and then to isolate and purify

Purifying

figure 7 .4: Men’s urine has high concentrations of male hormone. (Source: Alyce Santoro, for the author)

the hormone. By 1929, two groups (Doisy’s in St. Louis and Butenandt’s in Gottingen)49 had succeeded in crystallizing the urinary hormone and analyz­ing its chemical structure. But was it really the same as the hormone made in the ovaries? The final proof came in 1936, when Doisy and his colleagues used four tons of sow ovaries to produce a few crystallized milligrams of chemically identical molecules.50 The urinary hormone and the ovarian factor were one and the same.

The isolation of the male hormone followed a similar track. First, scien­tists developed a method of assaying an extract’s strength—the number of centimeters of regrowth over a specified time period of a cockscomb after castration (expressed in International Capon Units—ICU’s for short). Then they had to search for an inexpensive hormone source. Again, they found it in cheap and ubiquitous pee. In 1931, Butenandt isolated milligrams of male hormone from 2^,ooo liters of men’s urine collected from Berlin’s police bar­racks (figure 7.4).

Scientists had found male hormones in testes and men’s urine, and female hormones in ovaries and the urine of pregnant women. So far so good; every­thing seemed to be where it belonged. But at the same time, other research was threatening to unravel Steinach’s (and Lillie’s) formulation that each hor­mone belonged to and acted in its respective sex, defining it biologically and psychologically. To begin with, it turned out that the male and female hor­

mones came in several molecular varieties. There wasn’t a single substance, but a family of chemically related compounds with similar, but not identical, biological properties. The two hormones became many.51 Even more bewil­dering, there were scattered reports of female sex hormones isolated from males. In 1928, nine such reports appeared. The gynecologist Robert Frank wrote that he found this news ‘‘disconcerting’’ and ‘‘anomalous,’’52 while an editorial in the Journal of the American Medical Association called the report of female hormone bioactivity in ‘‘the testes and urine of normal men’’ ‘‘some­what disquieting.’’53 So convinced was the editorial writer of the unlikelihood of such a finding that he (I presume the pronoun is correct) questioned the validity of vaginal smear tests, which had become the standard of measure­ment in most of the laboratories working in female hormone purification.54

But the shock of finding female hormone in the testes and urine of ‘‘normal men’’ paled in comparison to another finding, published in 1934. In an article variously described by other scientists as ‘‘surprising,’’ ‘‘anomalous,’’ ‘‘curi­ous,’’ ‘‘unexpected,’’ and ‘‘paradoxical,’’55 the German scientist Bernhard Zondek described his discovery of the ‘‘mass excretion of oestrogenic hor­mone in the urine of the stallion’’—that cherished mythic symbol of virility.56 In short order, others found female hormones where they ought not to be. In 1933, thirty-five such scientific reports appeared, followed the next year by another forty-four. The first report of male hormones in females appeared in 1931, and by 1939 had been confirmed by at least fourteen additional publi-

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cations.

Actually, the first report of cross-sex hormone action had appeared as early as 1921, when Zellner reported that testes transplanted into castrated female rabbits could cause uterine growth. But the full import of such work became apparent only when the hormones of one sex turned up in the bodies of the other. Not only did contrary sex hormones appear unexpectedly in the wrong sex: they also seemed able to affect tissue development in their opposite num­ber! By the mid-i93os it was clear that male hormones could affect female development and vice versa. The anatomists Warren Nelson and Charles Mer – ckel, for example, noted the ‘‘amazing effect’’ of an androgen in females. Ad­ministration of this ‘‘male’’ hormone stimulated mammary growth, enlarge­ment of the uterus, ‘‘a striking enlargement of the clitoris,’’ and ‘‘periods of prolonged estrus.’’58

At first, scientists tried to fit their findings into the old dualistic scheme. For a while they referred to the cross-sex hormones as heterosexual hor­mones. What did heterosexual hormones do? Nothing, some suggested. They’re just nutritional by-products with no connection to the gonads. (So suggested Robert T. Frank, who claimed that ‘‘all ordinary foodstuffs contain female sex hormone. An average-sized potato contains at least 2 M. U. [mouse units].’’)59 The further discovery that the adrenal glands could make hetero­sexual hormones provided brief relief for those who found their existence anxiety-provoking. At least the gonads themselves still functioned along strict gender lines, since cross-sex hormones did not originate with them!60 As an alternative to the nutritional hypothesis, Frank found the presence of female hormone in the bile ‘‘of great theoretical interest and is of importance in explaining the occurrence of [sic] female sex hormone reaction in the blood of males and in the urines [sic] of males.”61

Finally, some argued that the heterosexual hormones indicated a diseased state. Although the men from whom estrogen was extracted appeared to be normal, they might, perhaps, be ‘‘latent hermaphrodites.’’62 But given how widespread the findings were, that position was hard to maintain. All of which led to a crisis of definition: if hormones could not be defined as male and female by virtue of their unique presence in either a male or a female body, then how could scientists define them in a manner that would prove translat­able among different research laboratories as well as the pharmaceutical companies that wished to develop new medicines from these powerful bio­chemicals?