Cytosolic and nuclear progesterone receptors in normal and luteal-phase-defect endometrial cells.

Endometrial biopsies were taken from 93 infertility patients registered in the Reproductive Endocrinology Clinic at the Medical College of Ohio. These endometrial specimens were assayed for nuclear and cytoplasmic progesterone receptors. Only the data from 53 normoprolactinemic euthyroid and nonhirsute patients were considered. Eighteen patients were found to have luteal phase defect by two consecutive out-of-phase biopsies. The remaining 35 patients had in-phase biopsies and were used as controls. Both groups were similar in age and clinical characteristics. Although we noted that the luteal-phase-defect group had fewer nuclear and cytoplasmic progesterone receptors than did the control group, the difference was not statistically significant.

Pulse labeling of hydroxyproline and leucine in reproductive tissues: a model for assessment of degree of reactivity.

A prospective study evaluating tissue reactivity by light microscopic and pulse labeling-autoradiographic techniques is presented. One uterine horn of a Sprague-Dawley rat model was subjected to transection and reanastomosis using microsurgical techniques (control). The opposite horn of each was also cross-sectioned with the application of an absorbable topical hemostatic agent (Avitene) and then reanastomosed. After closure of the abdomen, the animals were randomly assigned to groups according to the planned day of sacrifice (days 3, 7, 14, 21, or 28). Autoradiographic techniques utilizing [3H]hydroxyproline or [3H]leucine as well as light microscopy were employed to determine tissue reactivity. The results demonstrated an increase in stromal (P less than or equal to .05) and smooth muscle (P less than or equal to .01) reactivity with [3H]leucine labeling, and an increase in smooth muscle (P less than or equal to 0.05) reactivity with [3H]hydroxyproline in the Avitene-treated groups. It is concluded that pulse labeling with tritiated leucine and hydroxyproline is an effective method of objective analysis of tissue reactivity in reproductive organs.

Bleeding and vulvovaginitis in the pediatric age group.

Vaginal bleeding and vulvovaginitis are common clinical problems in the pediatric patient. Careful attention to historical and physical findings usually can resolve the problem at the initial visit. Knowledge of the differential diagnoses--including bacterial, nonspecific origins, dermatologic, traumatic, foreign bodies, neoplastic, urologic, precocious pubertal, or exogenous steroid ingestion--allows the clinician to include all significant causes of vaginal bleeding in this age group.

Evolving concepts in the mechanism of steroid action: current developments.

The mechanisms of steroid action remain a poorly understood enigma. Although much effort has focused on the steroid receptor as a mediator of the steroid's effect in the cell, we are only beginning to understand the structure of steroid receptors. Development of monoclonal antibodies directed against both the steroid-binding "receptors" and receptor-associated proteins has allowed novel approaches to the problem. They were important in determining the nucleotide sequences of several receptor genes and subsequently the amino acid sequence of three receptors. Surprisingly, receptors contain amino acid sequences common to v-erb-A, a potentiator of oncogenic transformation. Two receptor-associated proteins have been found and their relationship to the receptors suggests the possibility of additional functions of receptors in addition to binding deoxyribonucleic acid. Thus the role of the receptor in the mechanism of steroid action is evolving from the "two-step mechanism" to one that includes the recently discovered receptor-associated proteins.

Triiodothyronine receptors in porcine granulosa cells.

Triiodothyronine has been found to enhance gonadotropin- and insulin-stimulated morphologic luteinization and progesterone production by porcine granulosa cells in culture. The role of triiodothyronine in the above events is not well defined. One possibility is that triiodothyronine effects are direct and are receptor mediated. To test this concept, we undertook this study to investigate the presence of triiodothyronine receptors in granulosa cells aspirated from large (6 to 12 mm), medium (3 to 5 mm), and small (1 to 2 mm) porcine ovarian follicles. Crude nuclei were isolated and tested for iodine 125-triiodothyronine specific binding. Binding was time and temperature dependent and maximal in cells from small follicles at pH 8. Scatchard analysis yielded a mean apparent dissociation constant of 5.5 X 10(-9) mol/L and a mean apparent total number of binding sites of 1.0 pmol/mg of deoxyribonucleic acid. Competition experiments revealed the following relative binding affinities: triiodothyronine greater than L-thyroxine greater than reverse triiodothyronine. Gonadotropins, prostaglandins, epidermal growth factor, and insulin did not compete with 125I-triiodothyronine for binding. We conclude that there are nuclear binding sites in porcine granulosa cells with characteristics expected of a triiodothyronine receptor which might mediate a direct action of triiodothyronine on granulosa cells.

A 59-kilodalton protein associated with progestin, estrogen, androgen, and glucocorticoid receptors.

Previous studies of the anti 8.5S progestin receptor monoclonal antibody KN 382/EC1 showed that it was specific for nontransformed progestin receptors. However, with different methods of tissue disruption and the use of protease inhibitors, we found that other nontransformed steroid receptors formed immune complexes with KN 382/EC1. Binding of the antibody to rabbit uterine estrogen, progestin, and androgen and liver glucocorticoid receptor systems was characterized by sucrose density gradient centrifugation, high-pressure liquid chromatography (HPLC), immunoadsorption, and immunoblotting. Immobilized KN 382/EC1 adsorbed both Mr 59,000 and Mr 92,000 proteins. The Mr 92,000 protein appeared to be bound to the antigenic Mr 59,000 protein, and the two proteins were present in apparently the same stoichiometric relationship in several tissues. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoadsorbed material revealed appreciable amounts of both proteins in testis, stomach, lung, liver, uterus, and kidney. Only trace amounts were found in skeletal or heart muscle, and none was found in blood serum. Cleveland digestion of isolated Mr 59,000 and 92,000 proteins revealed dissimilar peptide constituents. Immunoblots of material from uterus and liver resulted in staining of the Mr 59,000 protein but not the Mr 92,000 protein. We conclude that similar antigenic determinants reside in components of several nontransformed steroid receptors and they reside on an Mr 59,000 protein. It is likely, therefore, that there are common components present in nontransformed steroid receptors.

Endometriosis in association with uterine anomaly.

Endometriosis is frequently a chronic process, which may begin soon after menarche. The process may be enhanced by mechanical obstruction. Theories of retrograde menstruation and metaplasia still remain in vogue. Endometriosis is a cause of both acute and chronic pelvic pain in the adolescent. We present case reports of müllerian lateral wall fusion defects with surgical correction and evidence for resorption of endometriosis. Clinicians must be aware that patients with uterine anomalies may develop extensive endometriosis, which upon creation of an unobstructed outflow tract results in complete resorption. Furthermore, the mechanism of formation of endometriosis in association with an outflow tract obstruction may be very different from that associated with infertility. We recommend consideration of endometriosis and/or a reproductive tract abnormality in the adolescent with persistent pelvic pain.