Firyal S. Khan-Dawood

Depot leuprolide acetate versus danazol for treatment of pelvic endometriosis: changes in vertebral bone mass and serum estradiol and calcitonin *

OBJECTIVEnTo determine changes in trabecular vertebral bone mass, serum E2, and serum calcitonin during and after therapy of pelvic endometriosis with depot leuprolide acetate (LA) or danazol.nnnDESIGNnProspective, randomized, double-blind study.nnnSETTINGnAcademic university hospital and department of obstetrics and gynecology.nnnPATIENTSnTwelve women with symptomatic pelvic endometriosis diagnosed and staged by laparoscopy.nnnINTERVENTIONSnAll patients received blinded treatment with either 3.75 mg JM depot LA given every month and daily placebo tablets (n = 6) or 800 mg oral danazol daily with a monthly placebo injection (n = 6) for 24 weeks.nnnMAIN OUTCOME MEASURESnQuantitated computerized tomography of bone density of thoracic 12 to lumbar 4 vertebral bodies were determined before, at the end of 24 weeks of treatment, and 6 and 12 months after completing treatment. Gain or loss of bone mass was based against pretreatment levels. Serial serum levels of E2 and calcitonin before, throughout, and after therapy were compared with changes in bone mass.nnnRESULTSnBone loss with LA was 14.0% +/- 0.5% (mean +/- SEM), recovering to a deficit of 4.2% +/- 3.8% and 3.3%, 6 and 12 months after stopping therapy. Danazol increased bone by 5.4% +/- 2.2%, with a further gain to 8.2% +/- 3.5% and 7.5%, 6 and 12 months after stopping treatment. Serum E2 levels usually were < 25 pg/mL (conversion factor to SI unit, 3.671) with LA but > 47.3 pg/mL with danazol. Calcitonin levels did not change significantly with either treatment.nnnCONCLUSIONnDepot LA produced marked sustained hypoestrogenemia and significant bone loss with incomplete recovery 1 year after stopping treatment. Danazol maintained normoestrogenemia and increased bone mass with the gain maintained even 1 year after stopping therapy.

Clinical, endocrine, and metabolic effects of two doses of gestrinone in treatment of pelvic endometriosis.

OBJECTIVEnOur purpose was to determine and compare the efficacy and hormonal and metabolic effects of 1.25 mg with 2.5 mg of gestrinone given twice a week in the treatment of mild and moderate pelvic endometriosis.nnnSTUDY DESIGNnA phase II, prospective, randomized, double-blind study involving 11 patients given gestrinone 1.25 mg (five patients) or 2.5 mg (six patients) orally twice a week for 24 weeks was performed. Revised American Fertility Society scores were determined by laparoscopy before and at the end of treatment. Serum hormone (free thyroxine, free testosterone, estradiol, progesterone, follicle-stimulating hormone, luteinizing hormone), sex hormone binding globulin, and lipid concentrations were measured before, throughout, and for 6 months after treatment. Quantitated computerized tomography of thoracic 12 through lumbar 4 vertebral bodies were determined before, at the end of, and 6 months after treatment.nnnRESULTSnGestrinone 2.5 mg significantly reduced the endometriosis implant score from 10.3 +/- 2.8 to 3.8 +/- 0.8 (p = 0.05). Both doses significantly reduced serum progesterone and sex hormone binding globulin levels. Estradiol, free testosterone, free thyroxine, follicle-stimulating hormone, and luteinizing hormone levels were not significantly affected. Spinal bone increased significantly by 7.1% with 2.5 mg but lost significantly by 7.1% with 1.25 mg gestrinone; these changes had not reversed completely 6 months after stopping treatment.nnnCONCLUSIONSnIn mild to moderate pelvic endometriosis 2.5 mg of gestrinone twice a week was more effective and had a more positive effect on bone mass than did 1.25 mg of gestrinone.

Oxytocin receptor and its messenger ribonucleic acid in human leiomyoma and myometrium.

OBJECTIVEnThe study determined the expression of oxytocin receptor and its gene in human uterine leiomyoma compared with the adjacent myometrium.nnnSTUDY DESIGNnPaired samples of leiomyoma and the adjacent myometrium from 20 women through the menstrual cycle, menopause, and various hormone treatments were studied. Oxytocin receptor was immunohistochemically localized with use of the specific antibody (2F8) to human oxytocin receptor. Oxytocin receptor protein was determined by Western blotting, whereas reverse transcription-polymerase chain reaction was used for oxytocin receptor messenger ribonucleic acid expression.nnnRESULTSnImmunohistochemistry showed positive staining in all tissues examined, relatively more intense in the myometrium than in the adjacent leiomyoma, and in tissues from the preovulatory than the postovulatory phase. Western blotting showed a single 70-kd band corresponding to the oxytocin receptor. The relative abundance of oxytocin receptor in both leiomyoma and myometrium was significantly higher during the preovulatory (n = 5) than the postovulatory (n = 5) phase (P = .034 and .05). In women receiving gonadotropin-releasing hormone agonist (n = 1) or oral contraceptives (n = 1), after the menopause (n = 2), and with irregular vaginal bleeding (n = 1), oxytocin receptor levels in leiomyoma and myometrium were unchanged but were reduced in anovulatory cycles (amenorrhea, n = 2). Reverse transcription-polymerase chain reaction showed messenger ribonucleic acid for oxytocin receptor as a 391-bp band in all leiomyomas and myometrium examined.nnnCONCLUSIONSnLeiomyoma and myometrium express the gene and protein for oxytocin receptor, which is probably partially regulated by ovarian sex steroids during the menstrual cycle.

Luteal insufficiency: Correlation between endometrial dating and integrated progesterone output in clomiphene citrate-induced cycles

Midluteal phase endometrium was histologically dated with midcycle luteinizing hormone surge time in 29 cycles from 10 parous women during untreated cycles (control) and treatment with clomiphene citrate 50 mg and 150 mg daily on days 5 through 9. Integrated progesterone output for 7 days after luteinizing hormone surge calculated from the daily plasma progesterone levels was 66.6 +/- 9.8 ng/ml in the control group compared with 117.5 +/- 18.6 ng/ml for clomiphene citrate 50 mg treatment and 152.1 +/- 11 ng/ml for clomiphene citrate 150 mg treatment (p less than or equal to 0.05). Only one cycle (clomiphene citrate 150 mg) had an out-of-phase endometrium and a significantly reduced integrated progesterone output of 28 ng/ml. All other cycles showed synchronous endometrial maturation. We conclude that luteal insufficiency as a result of clomiphene citrate treatment in ovulatory women is infrequent and is more likely to be a result of functional outcome of a relative lack of luteal phase progesterone output.

Insulin-like growth factor I receptors in human corpora lutea

OBJECTIVEnTo determine the presence and binding characteristics of insulin-like growth factor (IGF) receptors in corpora lutea (CL) of spontaneous and clomiphene citrate (CC)-induced cycles and to identify any relationship between IGF receptors and cytosol progesterone (P) and 17 alpha-hydroxyprogesterone (17 alpha-OHP) levels.nnnDESIGNnWomen undergoing bilateral tubal ligation were divided into two groups. One group received no medication (controls) and the other took 50 mg of CC. Midluteal phase CL were recovered at tubal ligation for hormone and receptor analysis.nnnSETTINGnPatients were recruited from a university hospital setting.nnnPATIENTSnEleven fertile women 26 to 37 years of age requesting bilateral tubal ligation were studied.nnnINTERVENTIONSnFour women were given 50 mg/d of CC from days 5 through 9 of study cycle. Seven women did not take any medication. Minilaparotomy bilateral tubal ligation and luteectomy were performed 7 to 9 days after midcycle urinary (LH) surge.nnnMAIN OUTCOME MEASURESnInsulin-like growth factor receptor concentrations and binding characteristics and cytosol P and 17 alpha-OHP levels in individual CL.nnnRESULTSnOptimal binding for 125I-IGF-I with membrane fractions of homogenized CL was obtained with incubation at 4 degrees C for 16 hours. Specific binding (mean +/- SEM) was significantly higher in CC-treated (53.6% +/- 4.8%) than in control cycles (25.9% +/- 5.5%, P less than 0.001). Receptor concentrations were also significantly higher in CL from CC-induced (145.6 +/- 21.8 pmol/mg protein) than from control cycles (74.8 +/- 15.2 pmol/mg protein, P less than 0.02). Insulin-like growth factor receptor levels correlated with neither serum nor cytosol P and 17 alpha-OHP in CL from either cycles.nnnCONCLUSIONnSpecific IGF-I receptors are present in human CL of the menstrual cycle with higher concentrations present in CC-induced cycles. Thus IGF may express its action on luteal function through its receptors in CL.

Plasma insulin-like growth factor-I, CA-125, estrogen, and progesterone in women with leiomyomas*

OBJECTIVEnTo determine plasma levels of insulin-like growth factor-I (IGF-I), CA-125, estrone (E1), E2, and P in women with uterine leiomyomas compared with normal women.nnnDESIGNnWomen with leiomyomas were compared with normal women (control).nnnSETTINGnUniversity Department of Obstetrics and Gynecology.nnnPATIENTSnFifty-one premenopausal women with uterine myomas > 14 weeks gestation and 30 normal fertile women (controls) were studied. Peripheral blood samples were obtained before myomectomy or hysterectomy and during the nonmenstruating phase in the controls.nnnMAIN OUTCOME MEASURESnPlasma levels of E1, E2, P, CA-125, and IGF-I were determined by specific and sensitive RIAs and immunoradiometric assays.nnnRESULTSnPlasma IGF-I levels were 2,006 +/- 185 mU/mL (mean +/- SEM, n = 35) and 2,335 +/- 287 mU/mL (n = 16) in women with leiomyomas during the follicular and luteal phases, respectively, whereas the corresponding values for normal women were 1,702 +/- 120 (n = 30) and 1,774 +/- 239 mU/mL (n = 30). Similarly, plasma CA-125 levels were unchanged in women with leiomyomas (myomas: 18.8 +/- 2.4, 21.5 +/- 3.7 U/mL; normal: 15.9 +/- 1.5, 15.8 +/- 1.3 U/mL during follicular and luteal phases, respectively). Women with leiomyomas had plasma E1, E2, and P levels during the follicular phase (91.9 +/- 11.5 pg/mL; conversion factor to SI unit, 3.699; 94.6 +/- 19.0 pg/mL; conversion factor to SI unit, 3.671; and 1.5 +/- 0.4 ng/mL; conversion factor to SI unit, 3.180, respectively) and the luteal phase (105.8 +/- 11.2 pg/mL; conversion factor to SI unit, 3.699; 128.7 +/- 24.8 pg/mL; conversion factor to SI unit, 3.671; and 9.6 +/- 1.6 ng/mL; conversion factor to SI unit, 3.180) similar to normal women.nnnCONCLUSIONnPlasma levels of IGF-I, CA-125, E1, E2, and P are normal in women with leiomyomas.

E-cadherin and its messenger ribonucleic acid in periimplantation phase human endometrium in normal and clomiphene-treated cycles

OBJECTIVEnThe purpose of this investigation was to determine whether treatment with clomiphene citrate, which is estrogenic and antiestrogenic, affects the expression of the cell adhesion molecule E-cadherin in human periimplantation phase endometrium.nnnSTUDY DESIGNnFive healthy women were studied for two cycles each, a control and a treated (clomiphene 50 mg daily, days 5 through 9) cycle. A biopsy specimen of endometrial tissue was studied (8 to 10 days post luteinizing hormone surge) for immunohistochemical localization, Western analysis of E-cadherin with use of a highly specific monoclonal antibody to human E-cadherin, and determination of messenger ribonucleic acid for E-cadherin by reverse transcription-polymerase chain reaction by use of oligonucleotide primers specific to E-cadherin and amplifying a 432 bp fragment.nnnRESULTSnLuteal phase plasma progesterone levels were significantly higher in clomiphene cycles. E-cadherin was immunocytochemically present in endometrium of control and treated cycles with no apparent difference in staining intensity. Western blots revealed the presence of E-cadherin. It was relatively more abundant in clomiphene-treated than control cycles but not significantly different. The message for E-cadherin gene is expressed in endometrium of control (n = 5) and clomiphene cycles (n = 4).nnnCONCLUSIONSnE-cadherin and its gene transcripts are expressed in periimplantation phase endometrium and are not significantly affected by clomiphene treatment.

9 Paracrine regulation of luteal function

Summary The mechanisms controlling luteal function may involve factors that are produced both within the corpus luteum and outside the ovary. The process of luteal control appears to involve a series of molecular species, proteins, peptides, steroids and prostaglandins. Each of these factors may act independently or in concert modifying the actions of one another. The effect of GnRH on luteal function has not been completely examined and thus its significance is unclear. The neurohypophyseal peptides, oxytocin and arginine vasopressin, in combination with LH, prolactin, oestrogens and prostaglandins may play an important regulatory role on the corpus luteum.

Localization and expression of oxytocin receptor and its messenger ribonucleic acid in peri-implantation phase human endometrium during control and clomiphene-treated cycles☆☆☆

OBJECTIVESnThis study was undertaken to determine expression levels of oxytocin receptor and its gene in peri-implantation phase human endometrium during clomiphene-treated cycles compared with control cycles.nnnSTUDY DESIGNnOxytocin receptor and its messenger ribonucleic acid in peri-implantation phase endometrium during control and clomiphene-treated (50 mg days 5 to 9) cycles of 5 healthy fertile women were determined by immunohistochemical methods, Western blot analysis with monoclonal antibody against amino acids 20 through 40 of the extracellular N-terminal human oxytocin receptor, and reverse transcription-polymerase chain reaction with oligonucleotide primers to amplify the 391-base pair fragment of the oxytocin receptor gene.nnnRESULTSnOxytocin receptor and its messenger ribonucleic acid were expressed in human peri-implantation phase endometrial samples from both control and clomiphene-treated cycles. The receptor was localized predominantly in the epithelial cells and glands, with little or none detected in the stroma. Oxytocin receptor protein was separated out as a single 70-kd band by Western blot analysis; its relative abundance was significantly reduced during clomiphene-treated cycles. The messenger ribonucleic acid was detected in all endometrium during control and clomiphene-treated cycles, with greater expression during control cycles.nnnCONCLUSIONSnThe expressions of oxytocin receptor and its gene in luteal phase human endometrium suggest a functional relevance in modulation of biochemical changes for implantation.

In vitro microdialysis of baboon corpus luteum : effects of oxytocin on total and pulsatile progesterone secretion

Baboon corpora lutea (two each from the early, mid- and late luteal phases) were individually microretrodialyzed in vitro for 48 h, 12 h initial baseline, 12 h retrodialysis with OT (9 mU/h), 12 h without OT and 12 h with cAMP (5 mmol/h). Progesterone (P) was measured by a sensitive and specific radioimmunoassay in 10-min fractions of retrodialysates and analyzed for P peaks by PC-pulsar 3.0. Neither OT nor cAMP had any effect on the characteristics of P pulses. In early and late luteal phase CL, OT inhibited P secretion within 1 h of administration followed by increased P secretion late during OT perfusion. In midluteal phase, OT did not affect P secretion. In all CL, P secretion was sustained or further increased during the 12 h after stopping OT. cAMP also sustained baseline or stimulated P secretion. In contrast, OT either increased total P output/12 h (28 to 49% above baseline) with a further increase of 21% to 296% above baseline after stopping OT, or inhibited total P output by 4% to 13% percent with a further decline of 51% to 61% after stopping OT. Thus, while overall OT is luteotropic, its dual effect (initial inhibition followed by stimulation) suggests direct and indirect effects through paracrine-autocrine mechanisms.