Effect of steroid hormones and antihormones on hypothalamic beta-endorphin concentrations in intact and castrated female rats.

The aim of the present study was to evaluate the effects of estrogens and androgens on hypothalamic beta-endorphin (beta-EP) concentrations. Intact or castrated female rats were chronically (2 weeks) treated with estrogen (estradiol benzoate) and/or antiestrogens (clomiphene, cyclophenil or epimestrol), and with androgens (dihydrotestosterone or dehydroepiandrosterone sulphate) and/or antiandrogen (cyproterone acetate). A group of rats treated with vehicle were studied as comparison. The beta-EP concentrations were measured by radioimmunoassay on acidic extracts of rat hypothalami. The administration of clomiphene and cyclophenil significantly reduced hypothalamic beta-EP concentrations in intact rats, while both drugs or estradiol benzoate increased the peptide concentration in castrated rats. Both intact and castrated rats treated with epimestrol showed hypothalamic beta-EP concentrations higher than vehicle treated rats. The estradiol-induced increase of beta-EP was not changed by the concomitant administration of antiestrogens. The administration of dihydrotestosterone significantly decreased beta-EP concentrations in both intact and castrated female rats, while the treatment with dehydroepiandrosterone sulphate only slightly decreased beta-EP levels in intact female rats. The cyproterone acetate-chronically treated rats showed higher beta-EP concentrations than vehicle-treated rats and these changes were reversed by the concomitant addition of dihydrotestosterone or dehydroepiandrosterone sulphate. These results showed that estrogens play a positive role while androgens negatively influence the hypothalamic beta-EP concentrations in female rats, supporting the view that central beta-EP might be a target of gonadal steroid feedback signals.

[Clinico-echographic study of the ovulatory phase, spontaneous and induced, conceptional and non conceptional cycles]. / Estudio clínico-ecográfico de la fase ovulatoria, ciclos espontáneos e inducidos, concepcionales y no concepcionales.

We present the results obtained on 110 menstrual cycles of 87 patients; echography study of ovulation, immunologic LH determination, basal body register card and serial cervical mucus score, since -6 to 0 day. Echography study determined follicular diameter, endometrial bulk and the presence of liquid in the cul-de-sac. We considered only those patients in whom the disappearance of the follicle was observed 24 hours before; being this fact, the indicator of ovulation, related to all other variables. These patients presented spontaneous (47.3%), epimestrol (28.2%) or clomiphene induced (24.5%) cycles, and they turned out to be pregnant or not in that cycle, (34.6%, 25.8% and 37.0% respectively). It was 36 (32.7%) pregnant women in all the The three follicular diameter curves for pregnant cycles, were similar being the preovulatory follicular range 21.5 mm for spontaneous cycles, 18.9 mm for epimestrol induced cycles and 20.7 mm for clomiphene induced cycles. Conceptional range was from 15 to 22 mms, 15 to 22 mm, and 14 to 27 mms, respectively. We didn't observe free fluid in cul-de-sac in 5 pregnant cycles (13.9%). Most of positive LH, are about -2 and -1 (21% and 50%, respectively). There were one pregnant patient and 2 no pregnant patient with repetitive, negative LH. In 50% of patients who became pregnant, the Nadir was on -1 day. In spontaneous conceptional group, bad cervical score was not observed. The good cervical score period is longer on conceptional group than in those with epimestrol. On conceptional group, dissociation between best score day and the day of ovulations was not observed, fact that we observed in non-conceptional group.

Estudio clínico-ecográfico de la fase ovulatoria, ciclos espontáneos e inducidos, concepcionales y no concepcionales / Clinico-ecographic study of the ovulatory phase, spontaneous and induced cycles, conceptional and non conceptional

Se presentan los resultados de 110 ciclos menstruales de 87 pacientes que fueron estudiadas con seguimiento ecográfico de la ovulación, determinación inmunológica de LH, curva de temperatura basal y estudio seriado del score cervical, desde el día -6 al día 0; el seguimiento ecográfico comprendió la determinación del diámetro folicular, el grosor endometrial y la presencia de líquido libre en el fondo del saco de Douglas. Se consideran sólo aquellas pacientes en que la última observación del folículo fue hecha hasta 24 horas de su desaparición, siendo éste el hecho que indica la ovulación, relacionándose las demás veriables con ello. Estas pacientes presentan ciclos espontáneos (47,3%), inducidos con epimestrol (28,2%) o clomifeno (24,5%) y resultaron embarazadas o no en este ciclo, 34,6%; 25,8% y 37,0% respectivamente. Fueron 36 casos con embarazo (32,7%). Las tres curvas de diámetro folicular para ciclos con embarazo resultaron notablemente similares, siendo el tamaño folicular preovulatorio de 21,5 mm. para ciclos espontáneos, 18,9 mm para ciclos inducidos con epimetrol y 20,7 mm para ciclos inducidos con clomifeno. El rango en los ciclos concepcionales fue de 15 a 27 mm, 15 a 22 mm y 14 a 27 mm, respectivamente. En 5 ciclos con embarazo (13,9%) no se observó líquido libre en el Douglas. La mayoría de los LH (+) se agrupan en los días -2 y -1 (21% y 50% respectivamente). Hubo una paciente que se embarazó y 2 que no se embarazaron que tuvieron LH iterativamente negativa. En el 50% de las pacientes que se embarazaron el Nadir térmico fue el día -1. En el grupo espontáneo concepcional no se observaron malos scores cervicales. El período de buen score es más largo en los grupos concepcionales espontáneos y con epimestrol. En el grupo de ciclos con embarazo no se observó disociación entre el día del mejor score y la ovulación, hecho que sí se observa en los ciclos no concepcionales

[Effects of different methods of ovarian stimulation in adult rats]. / Efectos de diferentes métodos de estimulación ovárica en ratas adultas.

The in vitro fecundation programs normally use different ovarian stimulation agents. The purpose of the present study was to compare the effects of some of these agents on indices of ovulation and serum levels of estradiol, progesterone, and FSH. Treatments studied consisted of clomiphene citrate, PMSG alone and in combination with clomiphene citrate, pure human FSH and epimestrol. The data obtained show PMSG alone and PMSG plus clomiphene citrate to be the most effective treatments, in terms of number of oocytes harvested. No differences were noted between serum levels of oestradiol, progesterone and FSH.

Quantitative structure-activity relationships of estrogenic steroids substituted at C14, C15.

The uterotropic activity of thirty 3-methoxyestradiol derivatives is measured and discussed on the basis of X-ray crystallographic results and quantitative structure-activity relationship analyses involving hydrophobic substituent constants pi and f as well as steric parameters Pr and L. In addition, estrogenicity is compared to data of interceptive activity and receptor binding affinity. All the biological data exhibit a high degree of intercorrelation. 17 beta-Hydroxysteroids having 14 alpha configuration reveal a generally better capability of high-affinity binding than those being 14 beta configurated. Between the uterotropic activity and the hydrophobicity of C14, C15 substituents, statistically significant correlations are found which suggest a close contact between the steroidal D-ring subsite and the receptor protein (e.g. for 14 alpha steroids: log UDD = -0.996 pi -0.392; n = 9, r = -0.943, s = 0.235, t = -7.5, alpha less than 0.001). The hydrophobic nature of both 14 alpha and 14 beta medium-sized substituents employed is shown by QSAR regressions to exert a stronger influence than steric effects. Furthermore, there are indications to additional hydrogen bonding and steric repulsion phenomena. As to the receptor-binding models discussed in the literature, it is concluded that the receptor protein has a high conformational flexibility to accommodate very different drug structures all having the common phenolic ring A. But, if an appropriate spacing of steroidal key atoms is recognized by the receptor and, consequently, the steroid-receptor complex is formed, the binding is complemented by hydrophobic interactions also in the D-ring region.

Radio-immunoassay of salivary progesterone for monitoring ovarian function in female infertility.

Fifty-two women, aged from 25 to 41 years, with infertility due to chronic anovulation were admitted to the study together with 36 age-matched controls with proven ovulatory cycles. Paired plasma (3 ml) and whole unstimulated saliva (10 ml) samples were collected over a 30 day period, starting from the first day of a menstrual bleeding, in patients, and throughout the menstrual cycle, in controls. Salivary progesterone levels, measured in women with infertility, ranged from undetectable values to 16 pmol/l during the first, and from 36 to 98 pmol/l during the second half of the monitoring period. In eugonadal women the steroid levels ranged from 34 to 46 pmol/l and from 96 to 780 pmol/l during the follicular and luteal phases, respectively. The saliva/plasma progesterone ratio ranged from 0.58 to 2.71 p. cent and a good correlation between salivary and plasma levels was found at each time of monitoring. Many (86 p. cent) of patients, which were randomly allocated to a low- or high-dose epimestrol administration schedule, appeared to be sensitive to the drug, achieving, after therapy, salivary progesterone levels which were within the range of controls. Since correct assessment of luteal function in basal conditions and during therapy requires multiple steroid measurements, and since saliva can be obtained by non-invasive techniques, salivary assays represent an attractive alternative to plasma ones for monitoring ovarian activity, also during specific treatment.

Ultrasound evaluation of the ovarian hyperstimulation syndrome.

282 cycles in 106 women going to be artificially inseminated at the AIED Center in Rome have been studied by means of ultrasound monitoring in order to evaluate the incidence of the ovarian hyperstimulation syndrome. In none of the patients, either in pharmacologically stimulated cycles (82 in 36 women) or in non stimulated cycles (200 in 70 women) moderate or severe hyperstimulation syndrome was reported. Only few mild cases occurred. In the non stimulated group only five cases with mild hyperstimulation occurred (2.5%). 31 pregnancies have been obtained (15.5% of all studied cycles). In the stimulated group 11 cases with signs of mild hyperstimulation occurred (13.4%) and 19 pregnancies were obtained (23.1%) of the stimulated cycles). No twin pregnancy was observed. The only molar pregnancy reported occurred in this group. All signs of hyperstimulation--either in stimulated cycles or in non stimulated patients--regressed spontaneously in the same cycle and anyhow they were not detected by ultrasound monitoring in the subsequent cycle.

[Treatment of hypothalamically-induced primary amenorrhea with epimestrol]. / Zur Behandlung der hypothalamisch bedingten primären Amenorrhö mit Epimestrol.

Eleven patients with hypothalamic primary amenorrhoea, aged between 17 and 23 years, were examined. Each patient was given 10 mg epimestrol (E) orally over a period of 10 days followed by a 20-day pause. This regimen was repeated 6 times. Gonadotropin secretory patterns were investigated between 9 a.m. and 2 p.m. before, during (on the 5th day of treatment in the 4th cycle) and 3 weeks after termination of treatment. In addition LHRH double stimulation tests were carried out before and after treatment to evaluate the acute releasable pool of gonadotropins as well as the pituitary synthesis capacity. In 3 patients with low baseline gonadotropin levels no effect of E on the release of gonadotropins could be found. These patients also showed non-pulsatile secretion with low baseline gonadotropin levels and no response in the LHRH stimulation tests. Similar results could be observed in one patient with low baseline LH levels but FSH levels within the normal range. Variable results were found in 7 patients with both LH and FSH in the normal range: 3 women experienced menstrual bleeding during and/or after E treatment; 1 woman stated that spotting had occurred twice. These 4 patients all showed regular pulsatile LH secretion after cessation of E treatment; the duration of the LH pulses was 60 to 120 minutes. Some hypothalamic activity seems to be essential for a positive response to E in patients with primary amenorrhoea. This activity is characterized by a positive gestagen test as well as baseline gonadotropin levels in the normal range.(ABSTRACT TRUNCATED AT 250 WORDS)

[Rate of pregnancy in the treatment of sterility during spontaneous and therapy cycles]. / Schwangerschaftsrate bei Sterilitätsbehandlung in Spontan- und Therapiezyklen.

Serum hCG was determined in 231 infertility patients in the late luteal phase of spontaneous and therapeutic cycles. The aim was to establish whether the discrepancy between the high number of induced ovulation cycles and the far lower cycle-related pregnancy rate can be explained by subclinical abortion. In 16.7% of spontaneous cycles hCG was positive; in 16.8% of these cases clinical pregnancy subsequently occurred. The rate of biochemically detectable pregnancies was no higher under endocrine therapy. However, under treatment with clomiphene, epimestrol, prolactin inhibitors and glucocorticoids the rate of clinical pregnancies was almost twice as high, and after hMG/hCG therapy clinically demonstrable pregnancies developed from biochemically identified ones in 76% of the cycles. The investigations show that a stimulation therapy leading to ovulatory cycles after multifollicular development increases the clinical pregnancy rate without resulting in a higher rate of very early abortions.

[Induction of ovulation in 1985]. / L'induction de l'ovulation en 1985.

There are many methods that can be used to induce ovulation when there is a fault in ovulation in patients who have normal prolactin levels. These are: Bringing the weight to a normal level. Giving Clomiphene. Giving Tamoxifen. Giving cyclofenil and bromocriptine, which really have no more effect than giving a placebo. Giving gonadotrophins in a classical way. This is very useful where there is hypogonadic amenorrhoea but much less useful when the failure of ovulation occurs with normal gonadic function. It is accompanied by a risk of multiple pregnancies and of hyperstimulation, which should be monitored by ultrasound very strictly so that it cannot become too serious. The use of purified FSH which theoretically should be more adequate, at least in cases where the gonadic function is normal in spite of failure of ovulation. Pulsatile administration of LHRH, which in cases of hypothalamic amenorrhoea carries less total risk than giving gonadotrophins. Finally, wedge resection of the ovaries which is reversed for polycystic ovaries that are larger than normal in size, and allied methods. The first choice for hypogonadic hypothalamic amenorrhoea would seem to be the LHRH pump; and for failure of ovulation with normal gonadic function Clomiphene or Tamoxifen. When anti-oestrogens fail to correct these latter cases one can choose according to the case between gonadotrophins, choosing if possible pure FSH, and/or wedge resection. In the last resort in these cases the LHRH pump can be used. The frequent failure of these methods show that perhaps it is possible to create a hypogonadotrophic hypogonadism by giving agonists for a long time or antagonists to LHRH in such a way that a second attempt can be made to induce ovulation using gonadotrophins in better conditions of efficacy and safety.

[Indications for hormone therapy of female secretory sterility]. / Indirizzi di terapia ormonale della sterilità secretoria femminile.

Female dysendocrine sterility has displayed a statistical incidence of 3.4% since 1967 in Milan's fertility and sterility centres. It is always marked by clear-cut clinical situations. Of these, particular interest is attached to anovulation (62.4% of cases), both with the cycle and with anovularity, ovarian micropolycystosis (2.7%), both as Stein ovary and as micropolycystic ovary, disturbances of ovary endocrine secretion: lutein deficiencies (21.2%) in the form of both brief and inadequate luteal phase. Treatment is aimed at possibly discontinuous reinstatement of ovulation. Clinical and pharmacological experiments over the last twenty years have put forward many "inducers". Mention is made of four personal approaches: --clinical employment of homologous gonadotropins (hMG + hCG), sequentially rather than paired, when poor gonadotropin secretion accompanied by insufficient endogenous oestrogenic activity is the main feature. Investigation from June 1964 to December 1981, coupled with monitorisation and personalisation of the treatment, initially through daily checks of total and fractionated oestrogenuria, and in recent years preferably through plasma 17-beta oestradiol or urinary enzyme determinations, has given a different slant to the reported disadvantages of gonadotropic management: hyperstimulation frequent multiple pregnancies, frequent multiple miscarriages; --employment of GnRH or its analogues (indications virtually those for paired gonadotropins). Some uncertainties however, exist with regard to the contraceptive action displayed by the agonist and antagonist analogues at certain doses, and with regard to the antigonadic action GnRH appears to have, both in the depression of oestrogen and progesterone production and in the arrest of follicular maturation an ovulation; --a preference for clomiphene among the antioestrogens in cases of primarily hypothalamic dysfunction and in ovarian micropolycystosis, provided endogenous oestrogenic activity is within normal limits; --a preference for hypoprolactinaemic drugs (bromoergocriptine, lysuride) in PRL-dependency, marked solely by an appreciable increase in serum LTH, screened as functional by means of selective tests; --experimentation of epimestrol, mainly in cases of sterility due to lutein deficiency.

A comparative study of clomiphene and epimestrol on plasma progesterone, CBG, TBG and SHBG, and salivary progesterone levels.

The plasma levels of CBG, TBG, SHBG and progesterone, and salivary progesterone levels were measured in eight young ovulatory volunteers. After the control cycle four subjects received 50 mg/day of Clomiphene from days 5 to 9 of the first treatment cycle, and 10 mg/day of Epimestrol from days 5 to 15 of the second treatment cycle. The other four subjects received the treatments in reverse order. Between the two treatments there was one cycle without medication as a "wash-out" period. Plasma and saliva samples were obtained on days +6, +8 and +10 (day of LH peak was denoted by 0), always between 08.00 and 09.00 h. Statistical evaluation was done by means of an analysis of variance (ANOVA), and correlation coefficients were also calculated. Evaluation of data on effects of Clomiphene and Epimestrol on the plasma levels of CBG and salivary progesterone showed that Clomiphene induced a highly significant rise (p less than 0.001) in the CBG levels and decrease (p less than 0.001) in salivary progesterone levels, while no changes were seen following administration of Epimestrol. Both Clomiphene and Epimestrol treatments led towards higher plasma progesterone levels, those following Clomiphene administration being higher. Neither treatment induced significant changes in TBG or SHBG levels. It is concluded that Clomiphene induces significant elevations of CBG and decrease in salivary progesterone, which is thought to reflect the free progesterone fraction and may have significance in relation to a discrepancy between the ovulation and pregnancy rates following Clomiphene therapy.

[Follow-up checks of children born from induced pregnancy (author's transl)]. / Nachuntersuchung von Kindern aus induzierten Schwangerschaften.

The following results were obtained by the authors from follow-up checks of 102 children, aged between 2.5 and eight years, who had been delivered from induced pregnancies. - Developmental abnormalities were recorded from seven cases, in the context of paediatric examinations. Minimal neurological deviations were exhibited by 14 children. Moderate retardation was established in psychological tests from something between 25 and 30 per cent of all cases. Positive responses to electroencephalography, however, without any clinical symptoms were observed in 20 per cent of the probands. Neurological symptoms and moderate retardation were recordable primarily from children who had originated from pathological pregnancies. The above findings are likely to support the recommendation to continue treatment of functional sterility by means of induced ovulation. Yet, treatment of that kind should be restricted to women unlikely to be afflicted with pathological pregnancy, which however, cannot be safely forecast in all cases.