Kerry L. Cheesman

Alterations in progesterone metabolism and luteal function in infertile women with endometriosis

The concentrations of pregnanediol-3-glucuronide (PGD) and pregnanolone (PN) were measured in daily morning urine specimens from 66 infertile women (40 with varying degrees of endometriosis and 26 control subjects) and correlated with daily changes in basal body temperature (BBT) and with midluteal levels of serum progesterone (P). PN and BBT rose at midcycle in women with endometriosis, as expected, indicating secretion of some P at that time. However, PGD, the major endpoint of P metabolism, was delayed in its excretion. Endometrial biopsies were similarly delayed (out of phase) in women with endometriosis, and a significantly higher incidence of follicular luteinization was seen. It appears that while P secretion begins at midcycle, the bulk of P secretion is delayed, perhaps because of the process of follicular luteinization, and that a shortened functional luteal phase thus exists in women with endometriosis.

Relationship of luteinizing hormone, pregnanediol-3-glucuronide, and estriol-16-glucuronide in urine of infertile women with endometriosis

The concentration of luteinizing hormone (LH), pregnanediol-3-glucuronide (PGD), and estriol-16-Glucuronide (E3G) were measured in daily morning urine specimens from 53 infertile women. In 26 of 29 women with various degrees of proven endometriosis, two distinct midcycle peaks of LH were found 2 or 3 days apart. Patients with LH peaks separated by 3 days had significantly more severe endometriosis than those with a single peak. Maximum concentrations of E3G were found to be delayed until after the first LH peak in these patients, and PGD concentrations did not rise until the time of the second LH peak, making actual luteal function of shorter duration than normal. From the data on LH, it appears that an inappropriate hormonal feedback mechanism is operative in endometriosis.

Radioimmunoassay of pregnanediol concentrations in early morning urine specimens for assessment of luteal function in women

A radioimmunoassay has been developed that employs a readily available radioligand 3H-20alpha-hydroxy-4-pregnen-3-one for the determination of pregnanediol glucuronide in urine. The unextracted steroid is assayed directly after dilution of urine with the use of an antiserum produced to a thyroglobulin conjugate of pregnanediol glucuronide. The assay has been validated, and the ability to assess luteal function by measurement of the concentration of pregnanediol glucuronide in early morning urine specimens has been demonstrated.

Entrapment of a highly specific antiprogesterone antiserum using polysiloxane copolymers.

Antiprogesterone antiserum was entrapped within a polysiloxane copolymer prepared from a 3:1 mixture of tetraethoxysilane and 3-aminopropyltriethoxysilane monomers. 400 microliters of this monomer mixture entrapped 70 mg of the 140 mg of immunoglobulins which were added, and the protein could not be washed from the highly stable copolymer which formed. Approximately half of the entrapped antiprogesterone antiserum was found to retain progesterone binding capacity with an apparent Ka equal to that of free antiserum in solution and was insensitive to effects of pH between 3 and 7. These preliminary observations and the unique chemistry of polysiloxane polymer formation suggest that such polymers may be useful in the entrapment of proteins for a variety of applications.

Reanalysis of antisera specificities and binding characteristics of rat pituitary hormone assays

Rat pituitary hormone radioimmunoassays (RIAs) are widely used in reproductive research, yet data on specificity and binding characteristics of many of the antisera are not widely available. This report characterizes one set of rat antisera supplied by the National Institutes of Health (USA). Rat follicle-stimulating hormone (FSH) and thyrotropin-stimulating hormone (TSH) antisera appear specific, but TSH exhibited significant competition in the rat luteinizing hormone (LH) assay. In addition, statistically significant nonparallelism was demonstrable in all three assay systems. This creates further problems in characterizing antisera cross-reactivity and may make potency estimates for pituitary standards inaccurate.

Hormonal Responses to Exercise in Non-Athletic Women

Women who had a history of regular menstrual cycles and who had not participated in a regular exercise or sports program for at least six months were recruited for a study of the effects of initiation of an exercise program on ovulation. After a control menstrual cycle during which basal hormone levels and the occurrence of ovulation were assessed, exercise on a treadmill was begun on the first day of the second menstrual cycle; exercise was continued at 85% of maximum heart rate for three sessions per week of from 15 to 30 minutes. The effect of this program on ovulation, luteal function, and on levels of Cortisol, testosterone, prolactin and growth hormone measured 10 minutes after cessation of exercise was studied during the month of exercise and at the same times of day in pre- and postexercise control months. Initiation of the exercise program resulted in a shortening of the follicular phase of the menstrual cycle with no effect on luteal phase length or on progesterone levels or pregnanediol excretion during the luteal phase. Testosterone and growth hormone were significantly elevated after exercise by comparison with values obtained during the control months, but Cortisol and prolactin were not. Cortisol levels may have been measured too soon after exercise began to detect an elevation. However, the complete absence of a prolactin elevation may be the reason for the failure of the brief exposure to exercise of this intensity to affect ovulation.

Relationships between the Amount of Sleep, Stress, and Ovarian Function in Women

The incidence of ovulation was studied in a group of 33 normal young women by means of changes in basal body temperature (BBT) and by the pattern of pregnanediol glucuronide and LH concentrations in morning urine specimens. Blood was also drawn for progesterone and LH assays on 10 consecutive days of one cycle in each of 15 of these women. Perceived stress and hours of sleep were recorded daily. Ovulation occurred by hormonal criteria in only 22 of 33 cycles studied, and only 14 of the ovulatory cycles had a detectable midcycle rise in BBT. Anovulatory subjects had significantly less sleep than subjects in the ovulatory groups. The frequency of ovulation was not influenced by the stress of blood withdrawal, nor was it related to the level of perceived stress. However, the perception of stress was significantly reduced during the luteal phases of ovulatory cycles in subjects having a clear rise in BBT.

Ability of implants of polymer-entrapped antiprogesterone antiserum to absorb and deplete progesterone from serum of the pregnant rat**Presented in part at the Thirty-Seventh Annual Meeting of The American Fertility Society, March 14 to 18, 1981, Atlanta, Georgia; and supported by subcontract 203 from the Program for Applied Research in Fertility Regulation.

A highly specific antiprogesterone antiserum (APA) produced by immunization of sheep with an 11 alpha-hydroxyprogesterone hemisuccinate-thyroglobulin conjugate was purified, and the IgG fraction was entrapped within a polysiloxane matrix. The matrix immobilized APA but allowed penetration and binding of progesterone (P) to the APA. In this entrapped form APA implanted intraperitoneally in rats on the tenth day of pregnancy resulted in a decline in serum P from 50 to 12 ng/ml within 12 hours and to less than 2 ng/ml within 36 hours. Free serum P measured by equilibrium dialysis fell to less than 0.2 ng/ml at 36 hours. Concomitant with the decline in serum P was a rise in both serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) and eventual fetal resorption.