Assessment of Incremental Dosage Regimen of Combined Oestrogen-Progestogen Oral Contraceptive

Eighty-six women of proved fertility used an incremental dosage regimen of a combined oral contraceptive for a total of 570 cycles over one year. A daily tablet containing 50 μg of ethinyloestradiol and 50 μg D-norgestrel was taken for 11 days and a daily tablet containing 50 μg ethinyloestradiol and 125 μg D-norgestrel for the next 10 days. Withdrawal bleeding occurred during the tabletfree interval of seven days. The new preparation proved to be an efficient contraceptive, well tolerated, and with few side effects. Women who had gained weight while taking other oral contraceptives lost weight when they changed to the new preparation. The regimen allowed a significant reduction in the cycle dose of progestogen, and these results suggest that a further reduction in the cycle dose of both oestrogen and progestogen may be possible without losing contraceptive efficiency.     

Hormone treatment of the adult transsexual patient

Hormonal reassignment has two aims: (1) to reduce the hormonally induced secondary sex characteristics of the original sex and (2) to induce the secondary sex characteristics of the new sex. In Europe, cyproterone acetate is generally used to inhibit androgens in male-to-female transsexuals. Medroxyprogesterone acetate is an acceptable, though less effective, alternative. To induce feminization there is a wide range of oestrogens. Oral ethinyloestradiol is a potent and inexpensive oestrogen, but it may cause venous thrombosis. Oral 17beta-oestradiol valerate or transdermal 17beta-oestradiol is the treatment of choice. The goal of treatment in female-to-male transsexuals is to induce virilization, including a male pattern of sexual hair, a male voice and male physical contours, and to stop menses. The principal hormonal treatment is a testosterone preparation. Hormone-dependent tumours have been encountered and surveillance is necessary.     

Mifepristone for the prevention of breakthrough bleeding in new starters of depo-medroxyprogesterone acetate

Depo-medroxyprogesterone acetate (DMPA) is an effective injectable contraceptive with worldwide availability. However, it is associated with a high incidence of breakthrough bleeding (BTB) during the first 6 months of use which often leads to discontinuation. Mifepristone is a progesterone receptor antagonist that has been demonstrated to decrease BTB caused by the levonorgestrel subdermal implant (Norplant). The purpose of this study was to determine if mifepristone would decrease BTB in new starters of DMPA. Twenty regularly cycling women who were new starters of DMPA were randomized to receive 50 mg of mifepristone or placebo every 2 weeks for 24 weeks. Percent days of BTB and number of cycles with bleeding intervals > or =8 and > or =14 days were evaluated using daily bleeding diaries. Ovulation was determined by measuring thrice-weekly urinary metabolites of estrogen and progesterone. Endometrial concentrations of ER and PR were determined by immunohistochemistry. Mifepristone significantly decreased the percent days of BTB and the number of cycles with prolonged bleeding intervals when compared to placebo. No subject ovulated in either group. ER immunostaining increased and PR immunostaining decreased after mifepristone treatment. In conclusion, a 50 mg dose of mifepristone taken every 2 weeks decreases the incidence of BTB in new starters of DMPA. This effect may be due to modulation of endometrial estrogen and progesterone receptors.     

Characterization of cyclicity and hormonal profile with impending ovarian failure in a novel chemical-induced mouse model of perimenopause

4-Vinylcyclohexene diepoxide (VCD) causes early, gradual ovarian failure in mice because it specifically targets small pre-antral ovarian follicles. The period between loss of these follicles and ovarian failure is analogous to perimenopause in women. We sought to characterize the period of onset of ovarian failure in VCD-treated mice in regard to estrous cycle length and hormonal changes. Female C57Bl/6 mice (age, 28 days) were dosed daily for 15 days with VCD (160 mg/kg intraperitoneally) to cause early ovarian failure or with vehicle only (control animals). Cycle length was monitored by vaginal cytology. Plasma levels of 17beta-estradiol (E2), progesterone (P4), and follicle-stimulating hormone (FSH) in control and VCD-treated animals were measured during proestrus of cycles 1 through 12. Cycle length (mean, 5.8 days) did not differ between groups for cycles 1 through 4. In contrast, cycle length during cycles 5 through 12 was increased (mean length, 10.9 days; P < 0.05 versus control) in VCD-treated animals, which also showed an apparent increase in plasma FSH levels. Plasma E2 and P4 at proestrus did not differ between groups during any cycle. Ovarian failure in VCD-treated mice was confirmed by histological evaluation on day 156 after onset of dosing, whereas control animals were still cycling. Therefore, despite compromised cycle length in VCD-treated mice, peak ovarian steroid production in preovulatory follicles at proestrus is adequate. These results demonstrate that the VCD-treated mouse can serve as an appropriate model to mimic hormonal changes during the perimenopausal transition in women.     

Ovulation-inhibiting properties of Org OD 14

Org OD 14 has recently been shown to be an interesting new steroid for the treatment of menopausal women. In view of the importance of treatment of perimenopausal women, in whom ovulation might occur, the aim of the present study was to assess whether or not Org OD 14, administered orally in a daily dose of 2.5 mg for 21 days, inhibits ovulation. Sixteen healthy female volunteers, aged 20-34 years and with established ovulatory cycles, were studied during a control cycle and a treatment cycle. Daily measurements of the plasma levels of FSH, LH, E2, P and PRL were made. Endometrial specimens were obtained from nine of the volunteers between 23rd and 25th day of both cycles. The criteria for an ovulatory cycle were: (1) mid-cycle FSH, LH and E2 peaks; (2) criteria (1) followed by a rise in the P levels of greater than 10 ng/ml; (3) a luteal phase of at least 12 days; (4) biphasic behaviour of E2; and, (5) secretory endometrium on days 23-25 of the cycle. All control cycles were ovulatory. During the treatment the mid-cycle FSH, LH and E2 peaks disappeared, and P levels remained very low. PRL levels showed an occasional moderate rise in some of the volunteers. Endometrial specimens showed a secretory pattern during the control cycle, and different degrees of proliferation during the treatment cycle in all nine volunteers. It was concluded that Org OD 14 inhibited ovulation in all 16 volunteers.     

Hormonal synchronization of the menstrual cycles of pigtail macaques to facilitate biomedical research including modeling HIV susceptibility

Background Menstrual cycle synchronization of female pigtail macaques could prove an invaluable resource in studies of the reproductive tract, associated infections, and other potential research fields. We tested whether use of an oral progesterone and estradiol combination tablet could synchronize menstrual cycles following treatment discontinuation. Methods Daily desogestrel 0.075 mg and ethinyl estradiol 0.01 mg were administered orally to three pigtail macaques at visual onset of perineal sex swelling and were continued until all animals had received it for at least 45 days. The hormones were discontinued, and these three macaques and three controls were observed for menstruation and had blood progesterone and estrogen measured over an additional 2‐month period. Results All treatment animals showed spontaneous menstrual cycle synchronization for 2 months after menstrual cycling resumed. Conclusion Progesterone and estradiol combination therapy can be used in pigtail macaques to induce synchronized cycling that persists in the absence of on‐going hormone treatments.     

Hormonal contraception

The combination hormonal contraceptive pill consisting of a fixed ratio of a progestational compound to an estrogenic compound administered from the 5th to 25th day of the cycle approaches 100% in efficacy. Cycle control is excellent and is related to the mechanism of action. The progestogen plus estrogen of the combination pill inhibit the hypothalamic-anterior pituitary-ovarian axis, thus insuring almost complete inhibition of ovulation and endogenous steroid hormone biosynthesis. The lack of endogeneous hormone is not critical since the pill replaces the lost hormones particularly as to uterine endometrial stimulation so that the tissue is prepared for a bleed on treatment withdrawal. The carefully timed hormonal replacement ensures excellent cyclicity. Although suppression of ovulation may be continued for years, cessation of treatment is followed by a return to normal hormonal function and fertility.Variants of the combination treatment include a once-a-month pill and a once-a-month injectable preparation. Both formulations are based on combinations of a progestogen plus estrogen.A high degree of efficacy approaching that seen for the combination pill has been achieved with a sequential regimen. In this procedure an estrogen alone is administered for 5 to 15 days while a mixture of estrogen plus a progestational agent is administered for the balance of the 20 days of treatment. Withdrawal of treatment brings on a bleed in 2 to 5 days. The mechanism of of antifertility is similar to that of the combination pill.Contraceptive efficiency of a high order may be attained with a small continuous dose of a progestational agent. The pregnancy rate and cyclicity are acceptable but not as good as that of the combination or sequential regimens. The pure progestogen treatment has been adapted to a pseudo post-coital therapy where one sexual contact requires one pill usually within 1 to 3 hours of the event. Thus far a reasonable efficacy has been established but as much as 33% of the cycles may be less than 20 days in length.A Single injection of a progestogen can produce effective antifertility for 90 days. This treatment is efficacious but the early treatment periods may involve considerable irregular bleeding and after repeated use ammenorrhea may be a problem.A true post-coital treatment has not been established. However, for emergency use 4 to 6 days of treatment with high doses of estrogen is highly effective in preventing pregnancy. The mechanism may involve speeding of ova transport thus preventing implantation.     

Comparative trial of nafoxidine and ethinyloestradiol in advanced breast cancer: an E.O.R.T.C. study.

A randomized clinical trial of nafoxidine, a non-steroidal oestrogen antagonist, and ethinyloestradiol in postmenopausal patients with advanced breast cancer produced objective remissions in 31% of 49 women receiving nafoxidine and in 14% of 49 receiving ethinyloestradiol. The differences in remission rates was almost significant (0.05 less than P less than 0.10). Life-threatening complications were more frequent with ethinyloestradiol than with nafoxidine but the latter produced specific toxic reactions on skin and hair that may limit its practical usefulness. Synthetic oestrogen antagonists may occupy a privileged place in the treatment of breast cancer, and other representatives of this new class of compounds should be accurately assessed in randomized clinical trials.     

Effect of Oestrogen Therapy on Plasma and Urinary Levels of Uric Acid

Uric acid clearance studies were carried out on a low-purine diet in 22 trans-sexual men before and during oestrogen therapy for this condition (stilboestrol in 21 cases, ethinyloestradiol in one). Plasma uric acid fell in 15 of the subjects and urinary uric acid rose in 17 of 20 subjects in whom satisfactory collections were obtained. These changes are significant and it is suggested that hormonal influences are responsible for the known age and sex differences in plasma uric acid.     

Artificial induction of lactation in dry cows

Oestradiol, progesterone, cortisol and reserpine were utilized in treatment regimes for hormonal induction of lactation in dry nonpregnant cows with previous reproductive failures. Lactations were successfully induced in 44 out of 55 cows. The udders of cows in which lactation had been induced successfully were visibly enlarged after 7 days of oestradiol-progesterone treatment on experimental day 8-14 and hence before the administration of cortisol (day 18, 19 and 20). It was concluded that addition of cortisol to oestradiol-progesterone treatment does not improve the success rate of hormonal induction of lactation. The addition of reserpine to oestradiol-progesterone treatment improved the success rate of hormonal induction of lactation. The reduction of oestradiol-17 beta dosage from 60 mg to 30-40 mg day-1 with progesterone at 150 or 200 mg day-1 for the initial 7 days increased both milk yield and the success rate of the hormonal induction of lactation and eliminated pelvic fractures and excessive oestrous activity in treated cows.     

Induction of luteolysis in the mare with a prostaglandin analogue

Five mares were administered 0.5 to 2.0 mg of a prostaglandin analogue, RS 9390 (Syntex), during nine estrous cycles in February and March. Luteolysis as measured by peripheral plasma progesterone occurred in four cycles, transitory luteolysis following 0.5 mg RS 9390 in two cycles, while functional corpora lutea were not present in three cycles. In 8 out of 9 of these cycles the mares returned to estrus 1.5 to 5 days following treatment. It appears that RS 9390 can be used as a regulator of cycle length in mares.     

Orchidectomy versus oestrogen for prostatic cancer: cardiovascular effects.

One hundred consecutive patients aged up to 75 with newly diagnosed cancer of the prostate suitable for hormonal treatment were included in a controlled study of the cardiovascular effects of oestrogen versus orchidectomy. In all cases pre-existing cardiovascular morbidity was excluded. Of the 100 patients, 91 were strictly randomised to receive either oestrogen (n = 47) or orchidectomy (n = 44) and 9 (6 given oestrogen, 3 orchidectomy) either chose their own treatment (five cases) or had it selected for them by the urologist (four). Oestrogen was given in the lowest recommended dosage in Sweden--namely, as 160 mg polyestradiol phosphate intramuscularly every month for the first three months, then 80 mg monthly, plus ethinyloestradiol 1 mg by mouth daily for the first two weeks, then 150 micrograms daily. At entry to the study the two treatment groups showed no difference in demographic characteristics or conventional risk factors for cardiovascular disease. During the first year, however, 13 (25%) of the patients given oestrogen suffered major cardiovascular events as compared with none of the patients after orchidectomy. Patients in the oestrogen treatment group who did not have minor signs of atherosclerosis at entry to the study suffered a similar incidence of cardiovascular complications to those who did have these signs at entry. The substantially increased risk of cardiovascular complications in patients given oestrogen for prostatic cancer warrants careful consideration when choosing treatment for this disorder.     

Effects of short-term treatment with a combined oestrogen-progestin oral contraceptive on biliary lipids and cholesterol saturation index in young women

The effect of daily ingestion for 7 days of ethinyloestradiol (30 micrograms) plus DL-norgestrel [0.5 mg] (Eugynon-30) on the lipid composition of duodenal bile in 8 healthy young women was investigated from the fifth day after onset of menstrual bleeding. This treatment did not significantly affect the concentrations of cholesterol, phospholipid and total bile acids expressed as mmol/l, nor the mean molar percentage of phospholipid. However, the treatment caused a significant increase in the mean molar percentage of cholesterol which was accompanied by a significant decrease in the mean molar percentage of total bile acids. The cholesterol saturation index of the bile of 7 subjects was elevated after treatment while both serum cholesterol and testosterone were significantly reduced. The results show that administration to healthy young women, not previously exposed to oral contraceptives, with a low oestrogen-progestin preparation for only 7 days produces a more lithogenic bile, accompanied by a decrease in serum cholesterol and plasma testosterone concentrations.     

Upregulation of cystic fibrosis transmembrane conductance regulator expression by oestrogen and Bak Foong Pill in mouse uteri

Although cystic fibrosis transmembrane conductance regulator (CFTR) has been shown to be expressed in the female reproductive tract, its functional role in the uterus is not fully understood. The present study investigated a possible physiological role of CFTR by comparing the effects of 17beta-oestradiol and Bak Foong Pill (BFP), an over-the-counter Chinese medicine used for centuries for the treatment of various gynaecological disorders, on uterus size and the expression of CFTR in the uterus of ovariectomised mice using RT-PCR. Treatment of ovariectomised mice with 17beta-oestradiol (0.2 mg/kg, p.o.) for 12 days caused a significant increase in uterine wet weight compared to vehicle. However, treatment with BFP (3 g/kg, p.o.) for the same period failed to increase uterine wet weight, indicating a lack of direct oestrogen-like activity of BFP. Analysis of CFTR mRNA expression in the harvested uteri using RT-PCR showed that both 17beta-oestradiol and BFP induced an increase in CFTR mRNA expression in mouse uteri compared to levels observed in vehicle-treated animals. These results suggest that CFTR can be upregulated by oestrogen and BFP, however, the effect exerted by BFP does not seem to be mediated by direct oestrogen-like activity. Regulation of CFTR expression by both oestrogen and gynaecological medication BFP indicates an important role of CFTR in reproductive functions.     

Pharmacokinetics of the progesterone-containing vaginal tablet and its use in assisted reproduction

Natural progesterone, which is devoid of androgenic activity, is widely used in assisted reproduction for luteal and pregnancy support. The vaginal route has become the most established way to deliver natural progesterone because it is easily administered, avoids liver first-pass metabolism, and has no systemic side-effects. The vagina has a large potential for absorption, and through the 'uterine first-pass effect' vaginal administration results in higher uterine progesterone concentrations. We have investigated the pharmacokinetics of natural progesterone in the form of a vaginal tablet. A single dose of 100 mg resulted in a mean C(max) of 31.53 +/- 9.15 nmol/l with a T(max) of 6.92 +/- 3.12 h. The terminal half-life was 16.39 +/- 5.25 h. The pharmacokinetic data are discussed in relation to dose, age, and estrogen priming. Single-dose pharmacokinetics of 100 mg of progesterone vaginal tablets and gelatin capsules were evaluated over 24 h. Results indicated a similar mean T(max) of 6.92 +/- 3.12 and 6.23 +/- 6.57 h, respectively. However, a significantly higher C(max) was achieved by the vaginal tablet (31.95 +/- 9.15 and 23.85 +/- 9.57 nmol/l, respectively, P < 0.05). Continuous use of vaginal progesterone did not influence the hormonal, liver, or lipid profiles evaluated. There was no case of endometrial hyperplasia. The vaginal tablet was found to be well-tolerated, safe, and easily administered. In conclusion, progesterone-containing vaginal tablets have good pharmacokinetic properties and should be used for progesterone supplementation in IVF.     

Lordosis behavior in castrated male rats treated with estradiol benzoate or testosterone propionate in combination with an estrogen antagonist, MER-25, and in intact male rats

Lordosis behavior could be elicited by manual stimulation in castrated male rats after treatment with estradiol benzoate (15 μg for 10 days) or testosterone propionate (1 or 3 mg for 10 days). The effect was antagonized by treatment with the estrogen antagonist MER-25 (10mg for 10 days). Prolonged treatment with testosterone propionate (1 mg for 26 days) resulted in display of male (nine of ten rats) as well as female (seven of ten rats) sexual behavior. Eleven of 32 intact male rats (age 120 days) and 22 of 37 other intact males (age 75 days) displayed lordosis in response to manual stimulation without hormonal treatment. Seven intact males which showed lordosis without hormone treatment were injected with MER-25 (10 mg/day × 10 days) and lordosis was abolished in six cases. The results suggest that estrogen is involved in the regulation of lordosis behavior in TP-treated and intact male rats.     

Menstrual cycles in rhesus monkeys (Macaca mulatta) are unaffected by a single dose of the anesthetics ketamine and xylazine administered during the midfollicular phase at laparoscopy

To identify an anesthetic regimen that produces more complete relaxation and analgesia than ketamine hydrochloride (Ketaset®) alone, a combination of ketamine (15 mg/kg body weight) and the hypnotic xylazine (Rompun®, 0.33 mg/kg) was evaluated. Since the desired experimental application required that the anesthetic not interfere with normal hormonal events during the menstrual cycle, this combination administered on day 6 of the cycle was tested to determine whether hormonal surges, incidence of ovulation, or cycle length would be altered relative to the use of ketamine alone. In five of six animals, ketamine plus xylazine had no effect on the occurrence of timely surges of estrogen, luteinizing hormone (LH), or follicle-stimulating hormone (FSH), or on ovulation as determined by the presence of a corpus luteum at laparoscopy and normal serum concentrations of progesterone. There were no significant differences between the cycle during treatment and previous cycles in the same animal for length of the menstrual cycle (26.0 ± 2.3 [5] days; X? ± S.D. [n] or luteal phase (13.4 ± 2.4 [5] days). Likewise, these values did not differ from those of ten control monkeys treated with ketumine only on day 5 or 6 of the cycle (incidence of ovulation, 10/10; cycle length, 27.9 ± 1.8 [10]; luteal phase length, 15.1 ± 1.4 [10], P > 0.05). Patterns of circulating progesterone were not altered by the addition of xylazine anesthesia. These findings indicate that xylazine, given in the midfollicular phase, did not alter ovulatory events or menstrual cycle characteristics in rhesus monkeys. Ketamine plus xylazine apparently provides anesthesia appropriate for laparoscopy.     

Repeatability of preovulatory follicular diameter and uterine edema pattern in two consecutive cycles in the mare and how they are influenced by ovulation inductors

Follicular diameter is used as a guiding tool to predict ovulation in the mare. However, the great range in preovulatory follicular diameter makes prediction of optimal breeding time based on follicular diameter unreliable. Uterine edema pattern is also useful to determine the best time to breed, since intensity of edema tends to dissipate as ovulation approaches, however, not every mare follows this pattern. The aims of this study were to assess the repeatability of preovulatory follicular diameter and uterine edema pattern in two consecutive spontaneous cycles and to determine how induction treatments (hCG, PGF(2)alpha and GnRH analogues) influence them. Fifty-three mares were followed during two consecutive cycles and scanned three times a day from 2 to 3 days before ovulation. During the first cycle, mares had a spontaneous ovulation and in the consecutive cycle mares received either: (a) no hormonal treatment; (b) 1500 IU hCG; (c) 125-250 microg Cloprostenol or (d) 2.1 mg Deslorelin implant. Mares ovulated consistently from similar follicular diameters in two consecutive spontaneous cycles (r=0.89; P<0.000). All three induction treatments had a significant effect on reducing the preovulatory follicular diameter (P<0.005). Mares showed fair correlation in uterine edema patterns in both consecutive non-induced cycles (r=0.71; P<0.005). In conclusion mares in consecutive cycles ovulated from consistent follicular diameters. Follicular diameters recorded from previous ovulations can be relied on to predict the optimal breeding time in successive cycles especially in mares that ovulate from unusually small follicles.     

Histological analysis of fat body development and molting events during soldier differentiation in the damp-wood termite, Hodotermopsis sjostedti (Isoptera, Termopsidae)

The caste system of termites is well defined, with a high degree of polyphenism among colony members. Polyphenic caste characteristics are hormonally regulated, and juvenile hormone (JH) is particularly involved in caste determination, as is the case with many other social insects. In the present study, soldier differentiation in the damp-wood termite, Hodotermopsis sjostedti, was induced by treatment with a JH analog (pyriproxyfen) in order to establish the chronology of tissular modifications appearing in response to the hormone.The fat body is involved in the physiological events that prepare the insect for the molting transition. The development of the fat body started within three days after hormonal treatment, and it filled the entire abdominal cavity for about four days prior to the molt to presoldier, maintaining this state until the next molt to soldier. Fat body development was accompanied by the accumulation of protein granules in the cytoplasm, but these granules disappeared during the few days preceding the molt to presoldier. The timing of consumption of these storage proteins corresponded to the window of epidermal growth, which was conspicuous about 14 days after hormonal treatment, and synthesis of the new cuticle, which was initiated 10 days after treatment. We summarize the chronology of the histological events under hormonal control.     

Opiate antagonist treatment reinstates estrous cycles in middle-aged persistent-estrous rats

Middle-aged female rats display luteinizing hormone (LH) surge deficits and cycle irregularity followed by the onset of persistent estrus (PE). The central nervous system has been identified as a primary locus of failure in PE rats, but the particular neural elements involved have not been determined. The goal of the present study was to identify a role for endogenous opioid peptides (EOP) in age-related acyclicity by evaluating the effect of opiate antagonist treatment on vaginal cytology in PE rats. PE rats were administered, s.c., saline (SAL), naloxone (NAL) or naltrexone (NTX) once daily for 20 days, or repetitively on Day 1 and on successive proestrus days if cyclicity was resumed. Single NTX (50 mg/kg), but not NAL (2 mg/kg), treatment interrupted the PE state in almost half of treated animals. Daily or repetitive proestrus NTX (10 mg/kg) treatment interrupted PE more frequently, and many animals displayed repeated estrous cycles and ovulation. Afternoon LH surges were observed after initial NTX treatment in animals displaying PE interruption. This demonstration that LH surges and ovulatory cycles can be reinitiated in PE rats with NTX suggests that dysfunction in the 'brake' on EOP secretion during proestrus may be one of the neuroendocrine impairments mediating acyclicity in aging female rats.