Carolyn J. Kubik

Effects of exogenous estrogen on uterine leukocyte recruitment

OBJECTIVE To determine the role of estrogen in leukocyte recruitment to the human endometrium. DESIGN Prospective, controlled in vivo study. SETTING Academic research laboratory. PATIENT(S) Ten patients presenting for donor oocytes. INTERVENTION(S) Endometrial biopsies for the evaluation of leukocyte populations were collected from perimenopausal women in two consecutive regulated cycles who were given two different regimens of estrogen with identical progesterone treatment. MAIN OUTCOME MEASURE(S) Immunohistochemical identification of endometrial leukocyte populations and relative levels of expression of three chemokine genes. RESULT(S) The total uterine leukocyte population increased significantly when the women received oral estrogen, which resulted in higher serum estrogen levels. This rise in leukocytes was due to a significant increase in both the uterine natural killer cells and the macrophage populations. T-cell numbers did not change relative to circulating estrogen levels. The relative abundance of mRNA from three chemokines was also determined. No changes were found in the expression of M-CSF or MCP-1. Interleukin 8 decreased in glands relative to estrogen levels. CONCLUSION(S) These data demonstrate that changes in circulating levels of estrogen can regulate the recruitment of bone marrow-derived cells to the uterine endometrium; however, the mechanism whereby that occurs remains elusive.

Randomized, prospective trial of leuprolide acetate and conventional superovulation in first cycles of in vitro fertilization and gamete intrafallopian transfer.

Ovarian stimulation after pituitary suppression with gonadotropin-releasing hormone agonists (GnRH-a) has been effective in women who have exhibited a poor response to conventional superovulation strategies. Their effectiveness in unselected women undergoing their first cycle of in vitro fertilization or gamete intrafallopian transfer, however, remains to be established. To address this question, we randomized 114 women to one of two treatment protocols. Protocol 1 consisted of 100 mg of clomiphene citrate on days 5 to 9, followed by 150 IU human menopausal gonadotropin (hMG) beginning on day 9. Protocol 2 consisted of daily GnRH-a beginning in the midluteal phase. Stimulation with 150 IU hMG commenced after pituitary down regulation and ovarian suppression were achieved. Human menopausal gonadotropin was continued in both protocols until adequate follicular development and serum estradiol concentrations were obtained. Protocol 2 patients reached egg retrieval significantly more often (87%) than Protocol 1 patients (61%), but the mean number of mature eggs retrieved and the pregnancy rate per retrieval were not significantly different between the two groups.

Endometriosis impairs the efficacy of gamete intrafallopian transfer: results of a case-control study*

OBJECTIVE To determine whether pelvic endometriosis impairs the efficacy of GIFT. DESIGN Matched follow-up study. SETTING University-based assisted reproduction program. PARTICIPANTS Patients undergoing GIFT between 1987 and 1991. Cases had a primary diagnosis of endometriosis. Controls had no endometriosis and were matched with cases according to age, number of mature eggs transferred, and sperm grade. INTERVENTION Gamete intrafallopian transfer was performed in all patients in an identical manner independent of their underlying diagnosis. MAIN OUTCOME MEASURES Pregnancy and delivery rates. RESULTS Of 114 laparoscopic egg retrievals performed in the endometriosis group, there were 37 pregnancies (32.5%) and 27 deliveries (23.7%). Of the 214 retrievals in the control group, there were 101 pregnancies (47.2%) and 76 deliveries (35.5%). Mantel-Haenszel estimates of relative risk indicated that endometriosis significantly impaired pregnancy and delivery rates. There was no statistically significant difference in pregnancy rates according to severity of disease among endometriosis cases. There was no statistically significant difference in pregnancy rates according to severity of disease among endometriosis cases. CONCLUSIONS Our finding that GIFT pregnancy rates were lower in women with a primary diagnosis of endometriosis than in matched controls suggests that endometriosis is associated with reduced efficacy of GIFT.

Ureteral obstruction as a complication of the Burch colposuspension procedure: Case report☆

Major complications of the Burch procedure for stress urinary incontinence are rare. Inadvertent kinking of the ureter during this procedure has been described only once previously in the literature. We present a second such case, hoping to draw attention to this rare but significant complication.

Establishment of pregnancies in humans after transcervical transfer of gametes immediately after oocyte retrieval

This study, like that of Veersema et al. demonstrates the feasibility of direct intrauterine transfer of gametes in initiating pregnancy. Whether the success of this procedure can be further improved with modification (preincubation, in vitro insemination) remains the subject of continued investigation. Notwithstanding this uncertainty, we feel that transcervical transfer of gametes may be an alternative to IVF for patients with ethical concerns regarding IVF.

Physiological range of human chorionic gonadotropin for support of early human pregnancy

OBJECTIVE To determine the physiological range of hCG in early pregnancy. DESIGN Retrospective study of patient charts. SETTING Magee-Womens Hospital IVF clinic, Monroeville, Pennsylvania. PATIENT(S) Sixty patients with successful, singleton birth outcomes. INTERVENTION(S) Serum hCG measurements on days 12-16 post-oocyte retrieval (OR). MAIN OUTCOME MEASURE(S) Lowest values, highest values, mean values, quartile mean values, and 48-hour doubling times for days 12-16 post-OR. RESULT(S) The average production of hCG in successful pregnancies is roughly 4-fold greater than the lowest amount observed in successful pregnancies, suggesting that a considerable excess of hCG is normally produced. Additionally, the average doubling time is almost 2-fold greater than the slowest doubling rate. CONCLUSION(S) The data from this study provide a set of values for the minimum and maximum threshold of hCG for days 12-16 post-OR that may be physiologically required, although not entirely predictive, for a successful IVF pregnancy outcome.

Effect of exogenous luteinizing hormone-releasing hormone on hypothalamic pituitary function in women with ovarian failure*†

Pituitary luteinizing hormone (LH) pulse frequency and amplitude were assessed as an indirect indicator of hypothalamic luteinizing hormone-releasing hormone (LH-RH) release in women with evidence of ovarian failure. Exogenous LH-RH (20 micrograms) was administered intravenously every 2 hours for 48 hours to determine the effect on pituitary LH release and the hypothalamic pulse generator for LH-RH secretion. This study design was used to investigate the possibility of an ultrashort negative feedback of LH-RH upon the synthesis and release of endogenous LH-RH. A wide range of LH pulse frequencies (8 to 28 pulses/24 hours) was present in these women. There was no discernible inhibition of hypothalamic LH-RH pulse frequency during or following exogenous LH-RH administration. Mean peripheral LH levels were significantly increased during exogenous LH-RH administration (P = 0.0038), reflecting both an increased baseline and an augmented pituitary LH response to the exogenous LH-RH. There were no differences found in LH pulse amplitude before and after LH-RH treatment. These data indicate that pituitary LH secretion in women with ovarian failure can be further stimulated by exogenous LH-RH. However, there was no evidence for inhibition of either pituitary LH secretion or hypothalamic LH-RH release using this administration schedule of exogenous LH-RH.

Enzymatic removal of asparagine-linked carbohydrate chains from heterodimer human chorionic gonadotrophin and effect on bioactivity.

The native form of human chorionic gonadotropin (hCG) is a heterodimer protein with two asparagine (Asn)-linked carbohydrate chains on each subunit. Removal of the Asn-linked carbohydrate chains from hCG has resulted in hCG variants with consistent antagonistic properties on isolated murine cells. Specific and direct enzymatic removal of these carbohydrate chains from native hCG with resultant antagonistic properties has not been reported. An antagonist to the hCG/luteinising hormone (LH) receptor could be used as an anticancer therapy, emergency contraceptive or for therapeutic resolution of ectopic pregnancies. Therefore, our aim was to use enzymes to specifically remove Asn-linked carbohydrate chains from hCG in the heterodimer form and analyse the resultant bioactivity. Native hCG was treated with endoglycosidases, carbohydrate removal was analysed with electrophoresis and the hCG variants were tested for altered bioactivity with human and murine cells. Endoglycosidases were able to cleave most of the Asn-linked carbohydrate chains from the native hCG. The deglycosylated hCG demonstrated a 75% reduction in bioactivity on a murine Leydig cell line and a 65% reduction in bioactivity on human granulosa cells. These results exemplify a simple and efficient method for creating deglycosylated hCG and provide the most direct evidence for the importance of Asn-linked carbohydrate chains in maintaining hCG bioactivity.

Estrogen Induces Premature Luteal Regression in Rhesus Monkeys during Spontaneous Menstrual Cycles, but Not in Cycles Driven by Exogenous Gonadotropin-Releasing Hormone

Administration of exogenous estradiol during the mid- to late luteal phase of the menstrual cycle results in premature regression of the corpus luteum. The present study was initiated to identify the site of action of estrogen as well as to determine why administration of estrogen during the early luteal phase of the menstrual cycle does not result in luteolysis. Based upon extant literature, we hypothesized that estrogen and progesterone synergize to promote premature luteal regression. We tested this hypothesis in intact, spontaneously cycling rhesus monkeys by inserting estradiol, progesterone, or estrogen plus progesterone capsules on days 2 through 6 of the luteal phase. Insertion of estrogen or progesterone capsules alone did not advance luteolysis compared with the effect of control empty implants (n = 3). In contrast, insertion of estrogen plus progesterone implants on days 2 through 6 of the luteal phase resulted in a significant lowering of serum progesterone concentrations, and menses was advanced 5-6 days compared with control cycles. On the basis of these findings in spontaneously cycling monkeys, we speculated that estrogen treatment causes luteal regression only in the presence of a progesterone-mediated decrease in LH pulse frequency. To test this hypothesis, we used rhesus monkeys whose endogenous gonadotropin secretion was abolished by either placement of radiofrequency lesions in the mediobasal hypothalamus or transection of the hypothalamic-pituitary stalk. Ovulatory menstrual cycles were restored by pulsatile administration of exogenous synthetic GnRH. Insertion of estradiol capsules during the luteal phase into animals whose gonadotropin pulse frequency was set at either one pulse per h or one pulse per 8 h failed to cause premature luteal regression (n = 4). These findings indicate that whereas estrogen promotes luteal regression in intact, spontaneous cycling rhesus monkeys, it does not do so in animals whose gonadotropin secretion is controlled by exogenous GnRH. On the basis of these observations, we conclude that the hypothalamus is a major site of action of estrogen in the initiation of luteal regression in macaques.