Ch. Bieglmayer

Treatment of skin ageing symptoms in perimenopausal females with estrogen compounds. A pilot study

A wide range of somatic symptoms of the perimenopausal female is due to the decrease of estrogen at that age. Minor attention has been paid hitherto to the involvement of estrogens in female skin ageing symptoms. In our study, the ageing skin of the face of perimenopausal females was treated with a 0.3% estriol cream (8 patients) or with a 0.01% estradiol cream (10 patients) for 6 months. Dermatologic follow-up was performed monthly. At each follow-up venous blood for radioimmuno assay determination of prolactin (PRL), follicle stimulating hormone (FSH) and estradiol (E2) was sampled. In addition, prior to and after 3 and 6 months of treatment, gynecological examinations for climacteric symptoms, mammary and colposcopic investigations and vaginal smears for cytology were performed. Both treatment groups showed improvement of the various skin ageing symptoms at the end of treatment. The effects of the group treated with topical estriol were slightly superior with regard to their extent and onset. No hormonal side effects were noted either clinically or by hormone monitoring. According to these preliminary results, local estrogen treatment appears to be a promising new approach for the treatment of skin ageing in perimenopausal females. However, for minimizing the risk of systemic hormonal side effects, concentrations and size of application field should be limited.

Relationship between the steroid and prolactin concentration in follicular fluid and the maturation and fertilizaton of human oocytes

Seventyeight follicles and their follicular fluid were aspirated from 46 women undergoing in vitro fertilization (IVF) procedures after stimulation of the ovaries with a low-dose human menopausal gonadotropin/human chorionic gonadotropin stimulation regimen. The concentrations of estradiol (E2), progesterone (P), testosterone (T), and prolactin (PRL) were measured in follicular fluid and related to the maturation of the oocyte-corona-cumulus complex (OCCC) and the fertilization of oocytes. Follicles containing mature oocytes had significantly higher follicular fluid E2 and P levels than follicles with intermediate and immature oocytes. A constant decrease in PRL and T values with advancing follicular maturation was observed. Similar results were obtained when the fertilizing ability of the oocytes was examined. The gradual decline in follicular fluid PRL and T levels during follicular development was connected with increasing E2 and P biosynthesis and therefore seems to be an important precondition for normal follicular and oocyte maturation.

OVULATION INHIBITION WITH DIFFERENT DOSES OF LEVONORGESTREL* AND OTHER PROGESTOGENS: CLINICAL AND EXPERIMENTAL INVESTIGATIONS

Abstract. Model systems were evaluated, which can be used to determine biological activities of progestogens at various cell levels of the anterior pituitary gland. in addition, various progestogens at different dose levels were tested in females to evaluate the ovulation inhibition dose. the association of progestogens with the cytosol receptor of the adenohypophysis of the rat was studied in vitro. Sucrose gradient centrifugation analysis revealed material sedimenting in the 4‐5 S and 6‐7 S region, resp., and both binding proteins were found to specifically bind progestogens. a small displacement by cortisol was observed in the 4‐5 S area. Relative affinity constants were estimated for various progestogens by incubation of cytosol samples with radioactive labeled progestins, and a close correlation between affinity and biological activity was recorded. Adult female rats seven days after castration were used to study the influence of various progestogens on LH‐RH stimulated gonadotropin release. Levonorgestrel was found to augment LH‐RH provoked LH release in vivo, whereas both basal as well as LH‐RH stimulated FSH serum levels were noted to be impaired at higher doses of the steroid. Similarly, δ15‐levonorgestrel was observed to increase the response of LH to LH‐RH. on the other hand, δ15‐levonorgestrel greatly augmented basal as well as LH‐RH induced FSH serum levels up to a dose of 10 μg. At 100 μg of δ15‐levonorgestrel FSH serum levels were suppressed again. But, only 1/3 of the levonorgestrel dose was necessary to produce these effects, i. e. δ15‐levonorgestrel was found ca. 3‐times more potent than the native progestogen. Experiments performed in a cell culture system of anterior pituitary cells revealed an impairment of LH‐RH stimulated gonadotropin release. LH and FSH release was suppressed to 40 and 50%, resp., at ED50 of LH‐RH in the presence of 3×10−8M levonorgestrel. LH and FSH release was reduced to 50 and 70%, resp., when anterior pituitary cells were incubated with LH‐RH in the presence of 3×10−8M δ15‐levonorgestrel. Thus, cell culture experiments also suggested that the potency of the δ15‐derivative of levonorgestrel was ca. 3‐times more potent. Analysis of patterns of cervical scores and karyopyknotic index as well as of serum levels of 17β‐estradiol, progesterone, LH and FSH revealed a borderline ovulation inhibition dose of levonorgestrel and δ15‐levonorgestrel at 50 and 30 μg, respectively. Cyproterone acetate, which is used in an oral contraceptive to treat acne and hirsutism was found to inhibit ovulation at a borderline dose of 1 mg. the present data combine to suggest that all model system are efficient to evaluate new progestogens, and that δ15‐levonorgestrel exerts a some 3‐times greater biological activity than levonorgestrel itself. Thus, a dose reduction of the progestogen component in oral contraceptives seems to be possible.

Influence of laparoscopic follicular aspiration under general anaesthesia on corpus luteum progesterone secretion in normal and clomiphene-stimulated cycles

Summary. In 32 patients with unstimulated normal cycles and 24 with cycles stimulated with clomiphene and human chorionic gonadotrophin (hCG) all visible follicles were punctured laparoscopically under general anaesthesia for the purpose of in vitro fertilization. In unstimulated cycles the time of surgery was between 24 and 32 h after the first luteinizing hormone (LH) increase in the urine; in the cycles stimulated with hCG (5000 i.u.) laparoscopy was between 35 and 37 h after injection. Blood samples for progesterone determination were taken about 7 days later. Progesterone levels were compared with those in a control group not subjected to surgery, in which the progesterone levels were determined 7 days after the LH increase. There was no statistically significant difference in the progesterone levels in the unstimulated subjects after laparoscopy compared with those in the control subjects but progesterone levels in the stimulated subjects were significantly higher (P<0·01). Durations of the luteal phases showed no significant differences thus laparoscopy under general anaesthesia does not impair luteal function.

Squamous Cell Carcinoma (SCC) Antigen in Patients with Invasive Cervical Carcinoma during Primary Irradiation

In a prospective study, serum concentrations of squamous cell carcinoma (SCC) antigen were determined by radioimmunoassay from 74 healthy volunteers and 54 patients with cervical carcinoma who underwent irradiation therapy. 5.4% of the controls had SCC levels greater than 3.0 ng/ml, which was considered as upper limit of the normal range. 31/54 (57.4%) patients and 60% of the patients with SCC had elevated pretreatment levels. In all patients with pretreatment serum levels above 3.0 ng/ml, SCC serum levels decreased during irradiation therapy. 4/5 patients with posttreatment levels greater than 0.5 ng/ml developed recurrence or persistence of tumor, 1 patient could not be followed up. Good conformity was found between SCC antigen serum levels and therapy response. SCC antigen determinations during and after therapy provide a useful tool in detecting progression and persistence of tumor.

Serum squamous cell carcinoma antigen levels in women with neoplasms of the lower genital tract and in healthy controls

SummarySquamous cell carcinoma antigen levels in 74 healthy volunteers, 57 patients with CIN and 91 patients with cervical carcinoma were determined by radioimmunoassay. 5.4% of healthy volunteers were above and all patients with CIN were below 3.0 ng/ml. 63.1% of 65 patients with primary squamous cell carcinoma, 1 out of 7 adenocarcinomas and 68.4% of 19 patients with recurrence of squamous cell carcinoma of the cervix had elevated SCC antigen levels. Elevated posttreatment levels carried a high risk factor of tumor persistence. Increases in SCC antigen levels during follow up usually signified recurrent carcinoma.

Ontogeny of 17β-estradiol-binding protein in the female rat hypothalamus and anterior pituitary

A line of evidence suggests that estrogen exerts a feedback on the hypothalamus-pituitary-axis. Experiments in the human indicate that two sets of estrogen sensitive neurones exist in the hypothalamus [ 11, of which one set is stimulated by estrogen to secrete LH-RH, whereas the other set of neurones secreting LH-RH is postulated to be sensitive to estrogen de& ciency. Similar mechanisms have been postulated for the rat [2-41. Previous observations [5,6] led to suggest that estrogen was an important factor involved in the maturation process of the brain-pituitary unit and in the regulation of the onset of puberty. Binding of 17fl-estradiol to the pituitary and brain was studied by autoradiographic techniques [7] and a specific, limited-capacity 17/I-estradiol-binding protein of the cytoplasm of the rat hypothalamus was described recently [8-131. Maximal responses to LH-RH were reported during pre-pubertal ages in rats [ 14,151 and changes were found in the in vitro responsiveness to LH-RH of pituitaries of rats at different ages [ 16,171. Furthermore, it was shown that 17/3-estradiol and other steroid hormones modulated LH-RH stimulated gonadotropin release [18,19]. The interaction of 17/3-estradiol with intracellular receptor proteins is an acknowledged primary event, which presumably mediates physiological activities in estrogen-responsive tissues. This process has been exten-

PROGESTERONE RECEPTOR IN THE RAT ANTERIOR PITUITARY Transformation and nuclear translocation

Cytoplasmic estrogen [ 1-3], glucocorticoid [3,4] and androgen receptors [3,5,6] have been detected in the rat pituitary. But, evidence for specific progesterone receptor has been conflicting [3,7]. Recently, progestin binding sites were detected in the rat anterior pituitary [8,9]. Previously, it was shown that progestins modulate according to their biological potency LH-RHstimulated gonadotropin release in vivo [ 10]. In addition, progesterone was observed to influence LH-RHstimulated LH and FSH release in vitro [11,12] and suppression of gonadotropin release is a well-acknowledged mechanism of action of hormonal contraceptives. These observations combine to suggest that progesterone acts at least partially at the pituitary level in regulating gonadotropin release. Whether or not specific progesterone receptors are present in the hypophysis is essential for the understanding of the mechanism of action of progesterone on the brain. Since estrogen was reported to induce cytoplasmic progesterone receptors in the animal uterus [ 13,14] it is likely that a progesterone receptor is detectable in neural tissues containing estrogen receptors. Ontogeny of the rat anterior pituitary estrogen-binding protein was described [15] and nuclear translocation of the estrogen receptor was reported [16]. The aim of the present investigation was to study the association of a progestin with the cytosol receptor in the rat anterior hypophysis and the transformation of the receptor complex in vitro. In addition, experiments were designed to investigate the nuclear translocation of the cytosol receptor complex in vitro. We were prompted to report these data since they add further evidence that receptor translocation is a primary step in the molecular events of progesterone action.

CA19-9, CA125 and CEA in Endometrial Carcinoma Tissue and Its Relation to Hormone Receptor Content and Histological Grading -

SummarySamples of 40 endometrial carcinomas were examined by immunohistochemical methods for CA19-9, CA125 and CEA. CA19-9 was detected in 93%, CA125 in 65% and CEA in 58%. CA19-9 was detected in more than 50% of tumor cells in 14 cases and the same was true for CA125 in six cases. In no tumor was CEA found in more than half the cells. The distribution of CA125 and CEA was markedly more heterogenous than that of CA19-9. There was no statistically significant correlation between immunohistochemical markers on the one hand, and estrogen and progesterone receptor content on the other. A correlation between histological grading and marker detection was only found for CA19-9. CA19-9 was detected in almost all endometrial carcinoma samples, and was the most homogenously distributed. This makes CA19-9 a possibly useful tumor marker for endometrial carcinoma.

Hormonal parameters in follicular fluid and the fertilization rate of in vitro cultured oocytes

SummaryOocytes and matched samples of follicular fluid were obtained from 52 preovulatory follicles aspirated laparoscopically for in vitro fertilization (IVF). Follicular fluid concentrations of estradiol (E2), progesterone (P), testosterone (T), prolactin (HPRL), prostaglandin F2α (PGF2α), prostaglandin E (PGE), protein content, and collagenolytic activity were measured and related to the fertilization rate of oocytes cultured in vitro. High concentrations of P and low levels of T and HPRL were associated with mature, fertilizable oocytes. Levels of PGF2α, PGE, and follicular fluid protein concentrations were similar in both groups. Mean collagenolytic activity was increased in the fertilized oocytes, although no significant difference could be observed. Our data demonstrate a close association between follicular fluid steroid and HPRL concentrations and successful fertilization of oocytes.