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Study Question: What is the effect of oestrogen and progesterone at the beginning of the menstrual cycle in delaying entry into the fertile window?, Summary Answer: Both oestrogen and progesterone contribute to a delay in the onset of the fertile window., What Is Known Already: Oestrogen enhances cervical mucus secretion while progesterone inhibits it., Study Design, Size, Duration: Observational study. Daily observation of 220 menstrual cycles contributed by 88 women with no known menstrual cycle disorder., Participants/materials, Setting, Methods: Women recorded cervical mucus daily and collected first-morning urine samples for analysis of oestrone-3-glucuronide, pregnanediol-3-alpha-glucuronide (PDG), FHS, and LH. They underwent serial ovarian ultrasound examinations. The main outcome measure was the timing within the cycle of the onset of the fertile window, as identified by the appearance of mucus felt or seen at the vulva., Main Results and the Role of Chance: Low oestrogen secretion and persistent progesterone secretion during the first week of the menstrual cycle both negatively affect mucus secretion. Doubling oestrogen approximately doubled the odds of entering the fertile window (OR: 1.82 95% CI=1.23; 2.69). Increasing PDG from below 1.5 to 4 µg/mg creatinine was associated with a 2-fold decrease in the odds of entering the fertile window (OR: 0.51 95% CI=0.31; 0.82). Prolonged progesterone secretion during the first week of the menstrual cycle was also statistically significantly associated with higher LH secretion. Finally, the later onset of the fertile window was associated with statistically significant persistently elevated LH secretion during the luteal phase of the previous menstrual cycle., Limitations, Reasons for Caution: This post hoc study was conducted to assess the potential impact of residual progesterone secretion at the beginning of the menstrual cycle. It was conducted on an existing data set because of the scarcity of data available to answer the question. Analysis with other datasets with similar hormone results would be useful to confirm these findings., Wider Implications of the Findings: This study provides evidence for residual progesterone secretion in the early latency phase of some menstrual cycles, which may delay the onset of the fertile window. This progesterone secretion may be supported by subtly increased LH secretion during the few days before and after the onset of menses, which may relate to follicular waves in the luteal phase. Persistent progesterone secretion should be considered in predicting the onset of the fertile window and in assessing ovulatory dysfunction., Study Funding/competing Interest(s): The authors declare no conflicts of interest. No funding was provided for this secondary data analysis., Trial Registration Number: N/A., (© The Author(s) 2024. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: The impact of testosterone (T) treatment on antidoping detection tests in female-to-male (F2M) transgender men is unknown. We investigated urine and serum sex steroid and luteinizing hormone (LH) profiles in T-treated F2M men to determine whether and, if so, how they differed from hypogonadal and healthy control men., Method: Healthy transgender (n = 23) and hypogonadal (n = 24) men aged 18 to 50 years treated with 1000 mg injectable T undecanoate provided trough urine and blood samples and an additional earlier postinjection sample (n = 21). Healthy control men (n = 20) provided a single blood and urine sample. Steroids were measured by mass spectrometry-based methods in urine and serum, LH by immunoassay, and uridine 5'-diphospho-glucuronosyltransferase 2B17 genotype by polymerase chain reaction., Results: Urine LH, human chorionic gonadotropin, T, epitestosterone (EpiT), androsterone (A), etiocholanolone (Etio), A/Etio ratio, dehydroepiandrosterone (DHEA), dihydrotestosterone (DHT), and 5α,3α- and 5β,3α-androstanediols did not differ between groups or by time since last T injection. Urine T/EpiT ratio was <4 in all controls and 12/68 (18%) samples from T-treated men, but there was no difference between T-treated groups. Serum estradiol, estrone, and DHEA were higher in transgender men, and serum T and DHT were higher in earlier compared with trough blood samples, but serum LH, follicle-stimulating hormone, and 3α- and 3β,5α-diols did not differ between groups., Conclusion: Urine antidoping detection tests in T-treated transgender men can be interpreted like those of T-treated hypogonadal men and are unaffected by time since last T dose. Serum steroids are more sensitive to detect exogenous T administration early but not later after the last T dose.

Luteinizing hormone (LH) is one of the main pituitary hormones that regulate ovulation, however its role has not been studied in giant panda. In this study, we developed an ELISA method for the detection of panda urinary LH. We analyzed urinary hormones of 24 female pandas during 36 breeding periods, we found females could easily be impregnated if the first mating occurred within 10 hours after LH peak. We also found the patterns of the ratios of urinary LH and progestagen in pandas that bred and successfully gave birth were significantly different from those that bred but failed to give birth. These data was the first to provide the urinary LH profiles during the estrous and gestational periods in pandas, and demonstrated that the appearance of the urinary LH peak indicated the timing of ovulation. The LH detection together with estrogen analysis makes the window for successful mating narrower than previously reported. Moreover, detection of urinary LH and progestagen can be used to discriminate between pregnancies and pseudopregnancies/miscarriages in the species. Thus, our findings suggest that LH not only plays a critical role in regulating ovulation but also plays an important role in maintaining pregnancy in the giant panda.

Objective: To compare daily sex hormone levels and rates of change between women with history of migraine and controls., Methods: History of migraine, daily headache diaries, and daily hormone data were collected in ovulatory cycles of pre- and early perimenopausal women in the Study of Women's Health Across the Nation. Peak hormone levels, average daily levels, and within-woman day-to-day rates of decline over the 5 days following each hormone peak were calculated in ovulatory cycles for conjugated urinary estrogens (E1c), pregnanediol-3-glucuronide, luteinizing hormone, and follicle-stimulating hormone. Comparisons were made between migraineurs and controls using 2-sample t tests on the log scale with results reported as geometric means., Results: The sample included 114 women with history of migraine and 223 controls. Analyses of within-woman rates of decline showed that E1c decline over the 2 days following the luteal peak was greater in migraineurs for both absolute rate of decline (33.8 [95% confidence interval 28.0-40.8] pg/mgCr vs 23.1 [95% confidence interval 20.1-26.6] pg/mgCr, p = 0.002) and percent change (40% vs 30%, p < 0.001). There was no significant difference between migraineurs and controls in absolute peak or daily E1c, pregnanediol-3-glucuronide, luteinizing hormone, and follicle-stimulating hormone levels. Secondary analyses demonstrated that, among migraineurs, the rate of E1c decline did not differ according to whether a headache occurred during the cycle studied., Conclusions: Migraineurs are characterized by faster late luteal phase E1c decline compared to controls. The timing and rate of estrogen withdrawal before menses may be a marker of neuroendocrine vulnerability in women with migraine., (© 2016 American Academy of Neurology.)

Objectives: Is the saliva test, Geratherm ovu control, as accurate as the established urinary luteinizing hormone (LH) test for detecting ovulation and the following the fertile period?, Study Design: The voluntary participants were 74 healthy women with regular menstrual cycles and not using any hormonal contraceptives. The women used Geratherm ovu control, a small plastic hand-held microscope, for detecting the fertile period. A drop of saliva from sublingual was put onto the lens of the microscope. Three results were possible: non-fertile (dot pattern), transitional and fertile (ferning pattern). The participants performed the saliva test from the 5th till the 22nd day of the menstrual cycle and noted the respective result in a table. In addition to Geratherm ovu control, the EXACTO test for determining urinary LH concentration and the time of peak fertility was also performed., Results: Positive LH shows a sharp increase beginning on the 10th cycle day with a maximum on the 17th cycle day. The curve for positive saliva and questionable positive saliva (one curve) is almost parallel with the curve for positive LH, reaching a maximum on the 16th cycle day. There is a high level of conformity for the same test results from the 5th (100%) till the 14th (84%) cycle day and from the 18th (80%) till the 22nd (96%) cycle day which corresponds to the pre- and post-ovulatory period., Conclusion: The saliva and the LH test both detect the fertile window of a menstrual cycle. Caused by the different hormones (estrogen for the saliva and LH for the LH test) leading to the respective positive test results, saliva turns positive 24h before LH. Consequently, the saliva test can be used as an ovulation test and help women maximize their chances of conceiving. There is also a high congruence between LH and saliva in the pre- and post-ovulatory period, indicating that the saliva test can also be used for contraception purposes., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

The aims of this study were to validate an enzyme immunoassay (EIA) for the measurement of luteinizing hormone (LH) in urine samples of Amazonian manatees (Trichechus inunguis; Mammalia: Sirenia) and to monitor urinary LH and reproductive steroids during the ovarian cycle in this species. Urine samples were collected from two captive males following a hormonal challenge with a gonadotropin-releasing hormone (GnRH) analogue. The urinary LH results from hormonal challenge were compared with urinary androgens for the purpose of EIA validation. Furthermore, urine samples were collected daily, over a 12-wk period, from two captive adult females, for 2 consecutive yr. The urinary LH pattern from females was compared with the patterns of urinary progestagens and estrogen conjugates throughout the ovarian cycle. An LH peak was observed in both male Amazonian manatees after the hormonal challenge, occurring prior to or together with peak androgen levels. In the females, the ovarian cycle ranged from 40 to 48 days (mean of 43.7 days). Two distinct peaks of estrogen conjugates were observed across all cycles analyzed, and the urinary LH peaks observed were accompanied by peaks of urinary estrogen conjugates. The EIA was validated as a method for the quantification of urinary LH from Amazonian manatees, as it was able to detect variations in the levels of LH in urine samples. These results suggest that T. inunguis exhibits a peculiar hormonal pattern during the ovarian cycle. Therefore, further studies are desirable and necessary to clarify the relationship between this hormonal pattern and morphological changes, as well as mating behavior, in Amazonian manatee.

Objective: It was hypothesized that aromatase inhibitor (AI)-induced interruption of estradiol negative feedback would modulate the reproductive hormone profile of obese women., Methods: Regularly cycling women aged 18-40 years with a BMI of 18-25 kg/m(2) (normal weight, n = 10) or >30 kg/m(2) (obese; n = 12) were given AI daily for 7 days. Urinary hormone profiles were compared between groups. Fourteen eumenorrheic, normal weight women not receiving AI stimulation served as historical controls. Urinary metabolites for LH, FSH, estradiol (E1c), and progesterone (Pdg) were measured and normalized to a 28-day cycle. Serum estrone and estradiol were measured in the late follicular phase., Results: Whole-cycle LH, FSH, and luteal Pdg excretion did not differ between obese (BMI = 37.1 + 7 kg/m(2) ) and normal weight women treated with AIs, although LH was greater in stimulated compared with unstimulated normal weight women. Whole cycle mean E1c was lower in AI-stimulated obese and normal weight participants compared with nonstimulated normal weight controls, but obese women treated with AI excreted far less E1c (467.7 ± 217.4 μg/mg Cr) than AI-treated normal weight women (911.4 ± 361.8 μg/mg Cr; P = 0.02). Follicular phase serum estrone and estradiol were also lower in AI-treated obese women versus AI-treated normal weight women (61.7 ± 22.8 and 18.3 ± 3.7 pg/ml versus 99.1 ± 30.5 and 37.7 ± 5.9 pg/ml, respectively; P = 0.034 and 0.005)., Conclusions: Normal gonadotropin output and luteal function occur at the expense of reduced E1c excretion in AI-treated women, and this discrepancy is particularly evident in obese women., (Copyright © 2014 The Obesity Society.)

Urinary steroid hormones and luteinizing hormone (LH) were monitored in a female pileated gibbon. The LH concentration almost peaked the day after estrogens peaks. The progesterone increased sharply after the LH peaks. Our results revealed for the first time the relationship between daily changes in urinary hormones throughout normal menstrual cycles in pileated gibbon., (© 2010 John Wiley & Sons A/S.)

Background: Alterations in circulating steroids are believed to be important mediators of the impact that diet and exercise have on breast cancer risk and changes in bone density. This study aimed to test the hypothesis that moderate exercise training combined with caloric restriction would produce significant menstrual disturbances and alterations in ovarian steroids in premenopausal women., Methods: Sedentary premenopausal women (25-40 years; body mass index: 23.6 +/- 0.6 kg/m(2)) assigned to either a light conditioning (LC, n = 9) or an exercise combined with caloric restriction group (EX + CR, n = 24) were studied for one screening, one baseline and four intervention periods equivalent to the length of subjects' menstrual cycles. Exercise consisted of supervised training sessions, i.e. two LC or four EX + CR times per week, 30-60 min at a moderate intensity. The EX + CR group was prescribed a diet representing a caloric restriction of 20-35% below baseline energy requirements, whereas the LC group remained eucaloric. Ovarian steroid exposure was determined with daily urinary estrone-1- and pregnanediol glucuronides (E1G and PdG, respectively) and mid-cycle urinary LH measures. Fitness, body composition, and serum sex hormone binding globulin (SHBG) and serum estradiol (E2) were assessed repeatedly., Results: The intervention produced significant increases in VO(2) max and decreases in both body weight (-3.7 +/- 0.5 kg; ranged from -8.8 to +1.8 kg) and percent body fat (-4.5 +/- 0.7%; ranged from -12 to +0.3%), which were attributable primarily to changes in the EX + CR subjects (time x group; P < 0.05). Serum E2 and urinary E1G and PdG concentrations declined significantly across the intervention period (time; P < 0.05), whereas SHBG increased transiently (time; P < 0.05) in the EX + CR subjects, with no significant changes observed in the LC group. The decrease in E1G area under the curve was significantly related to the daily energy deficit (R =0.61; P = 0.003), not the amount of weight lost. There was no significant impact of the intervention on menstrual cyclicity or the incidence of menstrual disturbances in either group., Conclusions: A moderate aerobic exercise training program combined with modest weight loss in accordance with recommended guidelines produces significant reductions in ovarian steroid exposure without disrupting menstrual cyclicity in premenopausal women aged 25-40 years. Exposure to a daily energy deficit is a stronger predictor of the decline in estrogen exposure than decreases in body weight.

Objective: To elucidate the reproductive hormone profiles in association with a diagnosis of diminished ovarian reserve (DOR)., Design: Prospective observational study., Setting: Academic tertiary care infertility practice., Patient(s): Eight regularly cycling infertile women diagnosed with DOR as the underlying contributor to infertility and 14 age-comparable healthy controls., Intervention(s): Daily morning urine voids were collected during one menstrual cycle., Main Outcome Measure(s): Urinary excretion of gonadotropins (FSH, LH) and metabolites of estrogen (E; estrone conjugate) and P (pregnanediol 3 glucoronide) during an entire menstrual cycle in women with DOR and healthy controls., Result(s): Women with DOR demonstrated significantly higher urinary FSH levels in the early follicular phase, exaggerated amplitudes, significantly protracted durations of LH, concomitant FSH surges, and significantly impaired luteal phase urinary excretions of estrone conjugate and pregnanediol 3 glucoronide compared with the controls., Conclusion(s): Women with DOR demonstrate reproductive hormone profiles that are distinct from age-comparable controls, share similarities with profiles previously described during menopause transition, as well as exhibit unique features not previously recognized in the context of reproductive aging., (Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

The cardiovascular consequences of hypoestrogenism in premenopausal women are unclear. Accordingly, the influence of menstrual status and endogenous estrogen (E(2)) exposure on blood pressure (BP), heart rate (HR), and calf blood flow in young (18-35 yr) regularly exercising premenopausal women with exercise-associated menstrual aberrations was investigated. Across consecutive menstrual cycles, daily urinary ovarian steroid levels were analyzed, and the area under the curve was calculated to determine menstrual status and E(2) exposure. BP, HR, blood flow, vascular conductance, and resistance were measured at baseline and following ischemic calf exercise. Exercising subjects consisted of 14 ovulatory (ExOv), 10 short-term (anovulatory and 100 days amenorrhea; LT-E(2) Def) E(2)-deficient women. Nine sedentary ovulatory subjects (SedOv) were also studied. All groups were similar in age (24.8 +/- 0.7 yr), height (164.8 +/- 1.3 cm), weight (57.9 +/- 0.9 kg), and body mass index (21.3 +/- 0.3 kg/m(2)). E(2)-deficient groups had lower (P < 0.002) E(2) exposure compared with ovulatory groups. Resting systolic BP, HR, blood flow, and vascular conductance were lower (P < 0.05) and vascular resistance higher (P < 0.05) in LT-E(2) Def compared with both ovulatory groups. Peak ischemic blood flow, vascular conductance, and HR were also lower (P < 0.05) and vascular resistance higher (P < 0.05) in LT-E(2) Def compared with all other groups. Our findings show that exercising women with long-term E(2) deficiency have impaired regional blood flow and lower systolic BP and HR compared with exercising and sedentary ovulatory women. These cardiovascular alterations represent markers of altered vascular function and autonomic regulation of which the long-term effects remain unknown.

Purpose: Exposure to residential magnetic fields may disrupt the normal nocturnal rise in melatonin levels, resulting in increased risk for breast cancer, possibly through increased levels of reproductive hormones. We investigated whether exposure to a 60-Hz magnetic field under controlled conditions is associated with a decrease in urinary nocturnal 6-sulfatoxymelatonin level and increase in luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estrogen levels in healthy premenopausal women., Methods: Using a crossover design, half the participants were assigned to magnetic field exposure of 5 to 10 mG greater than ambient levels for 5 consecutive nights during the early to midluteal phase of the menstrual cycle. On the last night of exposure, a nocturnal urine sample was collected. The next month, participants were sham exposed. The other half of participants were assigned the reverse order of exposure., Results: Magnetic field exposure was associated with decreased 6-sulfatoxymelatonin levels, but no changes in reproductive hormone levels were observed. Participants using prescription medications and anovulatory participants had more pronounced decreases in 6-sulfatoxymelatonin levels with magnetic field exposure., Conclusion: This study provides further evidence that exposure to magnetic fields is associated with decreased nocturnal melatonin levels, but does not support the hypothesis that such exposure results in increased urinary levels of estrogens, LH, or FSH.

Although reproductive aging has been separately related to elevated FSH and shorter follicular phase (FP), the direct association between both parameters has not been investigated. Also, the exact effects of increased FSH on estrogen production are yet to be established.A large database of daily urinary concentrations of FSH, LH, and estrone 3-glucuronide (E1G) from 37 regularly menstruating women (median 11 cycles per patient) was used. Initial FSH levels (iFSH) were estimated as the mean value of d 1-5. The day of E1G take-off (ETO) was determined by an algorithm, and accordingly, the FP was divided into early (d 1 to ETO) and late (ETO+1 to LH peak). FP maximum and integrated E1G were calculated. Subjects were distributed according to their mean iFSH into three categories (5 to 10, and >10 IU/liter). There was a gradual decrease in FP length with increasing category (15.2 +/- 3.8, 14.1 +/- 3.6, and 13 +/- 2.6 d, respectively; P < 0.0001). A similar effect occurred in early FP (7.5 +/- 4, 6.4 +/- 3.7, and 5.4 +/- 2.7; P < 0.0001); in contrast, late FP was unaffected (7.7 +/- 2.1, 7.7 +/- 2.1, and 7.6 +/- 2.4; P = 0.86). No consistent increase in E1G was found with advancing iFSH category; however, women with mean initial LH higher than 6 IU/liter had significantly elevated maximum (P < 0.0001) and integrated (P = 0.002) E1G.FP length decreases in parallel with increasing iFSH, with a selective effect on the early FP. Increased FSH does not affect E1G; however, elevated initial LH level was related to higher E1G.

The purpose of this study was to compare the fertile phase of the menstrual cycle as determined by the Clearplan Easy Fertility Monitor (CPEFM) with self-monitoring of cervical mucus. One-hundred women (mean age = 29.4 years) observed their cervical mucus and monitored their urine for estrogen and luteinizing hormone metabolites with the CPEFM on a daily basis for 2-6 cycles and generated 378 cycles of data; of these, 347 (92%) had a CPEFM peak. The beginning of the fertile window was, on average, day 11.8 (SD = 3.4) by the monitor and day 9.9 (SD = 3.0) by cervical mucus (r = 0.43, p < 0.001). The average first day of peak fertility by the monitor was 16.5 (SD = 3.6) and by cervical mucus 16.3 (SD = 3.7) (r = 0.85, p < 0.001). The mean length of the fertile phase by the monitor was 7.7 days (SD = 3.1) and by cervical mucus 10.9 days (SD = 3.7) (t = 12.7, p < 0.001). The peak in fertility as determined by the monitor and by self-assessment of cervical mucus is similar but the monitor tends to underestimate and self-assessment of cervical mucus tends to overestimate the actual fertile phase.

Objective: To monitor gonadotropin and ovarian hormone levels in relation to sleep duration in normally cycling women., Design: Observational and cross-sectional study., Setting: Multicentric collaborative study., Patient(s): One hundred six healthy and normally cycling women, aged 19 to 44 years, with cycle lengths of 24 to 34 days., Intervention(s): Follow-up during one to four consecutive cycles with daily urine collection., Main Outcome Measure(s): Urine concentrations of LH, FSH, estrone-3-glucuronide (E1-3-G), and pregnanediol-3-alpha-glucuronide (Pd-3alpha-G). Ultrasound determination of day of ovulation and estimation of sleep duration., Result(s): We found a significant association between FSH levels and sleep duration (P=.008). Follicle-stimulating hormone levels were 20% higher in long-time sleepers than in short-time sleepers. This association persisted whatever the age or the body mass index. There was no significant association between belonging to any group and LH, E1-3-G, or Pd-3alpha-G levels., Conclusion(s): Our results suggest that sleep duration could be related to FSH levels, although the study design did not allow us to establish a causal relationship nor to explain the physiological basis of the observed relationship.

Objective: To improve prediction of ovulation in normal cycles., Design: Collection of women's characteristics and their menstrual cycles. Monitoring and analysis of time relationships between several indicators of ovulation: transvaginal ultrasonography, cervical mucus, basal body temperature, urinary luteinising hormone, and ratio of urinary oestrogen to progesterone metabolites., Setting: Each of eight natural family planning clinics was to study 12 women for at least three cycles., Population: One hundred and seven normally fertile and cycling women aged 18 to 45., Methods: Daily measurements of urinary luteinising hormone, follicle stimulating hormone, oestrone-3-glucuronide and pregnanediol-3alpha-glucuronide. Basal body temperature recording and cervical mucus checking. Transvaginal ultrasound examination of the ovaries., Main Outcome Measures: Delays between the expected day of ovulation according to the luteinising hormone peak or to ultrasound evidence and the expected days according to the other indices of ovulation., Results: Ultrasonography was able to show evidence of ovulation in 283 out of 326 cycles. The average time lag between luteinising hormone peak and ultrasound evidence was less than one day (+0.46) but premature and late luteinising hormone-expected date of ovulation were observed in nearly 10% and 23% of cycles, respectively. Basal body temperature rise was observed in 98% of cycles. Cervical mucus peak symptom, rapid drop in the ratio of urinary metabolites, and luteinising hormone initial rise were all close to ultrasonographic evidence in more than 72% of cycles., Conclusions: For accuracy and practical reasons, the cervical mucus peak symptom, the ratio of urinary metabolites and luteinising hormone initial rise might be better indices of ovulation than the luteinising hormone peak.

To identify the developmental changes in monthly urinary gonadotropin and ovarian hormone excretion, consecutive 30-d first morning void urinary specimens were collected from 36 normal girls, one normal woman, and 15 female patients with idiopathic precocious puberty. Of these children, three normal girls and three patients with precocious puberty volunteered to collect these specimens on 2-3 occasions over a time interval of 0.5-3.2 y. When sampled, six were early prepubertal, nine late prepubertal, eight early pubertal, eight mid-pubertal, and eight late pubertal normal girls, and six were early pubertal and 14 mid-pubertal patients with precocious puberty. The mean level of monthly urinary LH, FSH, and total estrogen excretions increased with pubertal maturation. In prepuberty, the mean LH level was lower than the mean FSH level, and neither showed significant episodic fluctuations. In early puberty, mean FSH levels increased with remarkable fluctuations, and mean LH levels were low with few variations in the course of a month. At the onset of puberty, gonadotropin excretory patterns underwent specific changes, showing at the same time periodically and every other day fluctuating patterns. Urinary total estrogen and pregnanediol excretion fluctuated independently from these periodic variations in urinary gonadotropins. These patterns were observed in six out of 16 patterns in normal pubertal girls and 10 out of 20 patterns in precocious puberty. Once the urine LH level exceeded the urine FSH level, however, these periodic variations disappeared. The cycle of a normal postmenarcheal girl aged 14 y showed a pattern similar to that of a normal adult. In patients with precocious puberty, the hormonal patterns were similar to those of sexual stage-matched normal girls.

We have developed a method of estimating day of ovulation using urinary ovarian hormone data. The method identifies a day of luteal transition that occurs at the shift from production of follicular oestrogen to luteal progesterone. The algorithm for identifying this shift was evaluated and judged better than specified alternatives in that it resulted in (1) a high concordance between the day of luteal transition and peaks in urinary luteinizing hormone (LH) for cycles with well-defined peaks, (2) a low variance in the length of the luteal phase of the menstrual cycle, which presumably reflects a low measurement error in estimating day of ovulation, and (3) a high proportion of cycles for which an approximate day of ovulation could be determined. To validate the new algorithm, it was applied to an independent data set. The algorithm identified a day of luteal transition in 88 per cent of these cycles, and the identified day occurred within two days of the urinary LH peak for all the cycles with clear LH peaks. Determination of the day of luteal transition to estimate ovulation requires only first-morning urine specimens, requires no correction for day-to-day variations in urine concentration, and can be applied to a mid-cycle window of data.

The concentrations of various estrogen glucuronides and pregnanediol-3 alpha-glucuronide have been measured in daily samples of early morning urine (EMU) and 24 h pooled collections throughout a total of 70 menstrual cycles (58 subjects). The immediate objective was to identify and assess the potential value of measuring hormone metabolites to locate the fertile period in women, with the ultimate aim of developing a non-invasive immunochemical test. The project was undertaken in five centres and the results showed that: (i) there was good agreement in the patterns and mean concentrations of metabolites between the centres; (ii) estrone-3-glucuronide should be the estrogen glucuronide selected for further study; (iii) the analysis of EMU provided information that was as useful as the corresponding measurements on pooled collections for 24 h; (iv) a rise in the concentration of estrone-3-glucuronide (50% over the mean of three preceding values) could be used to locate the start of the fertile period in greater than 90% of the ovarian cycles (from day LHmax -- 3 to day LHmax -- 7); attempts to locate the end of the fertile period by a threshold value or a defined rise in the level of pregnanediol-3 alpha-glucuronide were disappointing. A 50% rise over the mean of three preceding values in the derived concentration ratio of estrone-3-glucuronide/pregnanediol-3 alpha-glucuronide could be used to indicate the start of the fertile period, and a 50% decrease plus two days the start of the infertile period in greater than 80% of the ovarian cycles. The limitations, implications and future developments are discussed.

We have evaluated a new micrometering kit using hemagglutination inhibition reaction for determination of total estrogens in urine of normal fertile women and women undergoing in vitro fertilization and embryo transfer (IVF-ET). The micrometering kit was sensitive enough to monitor follicular maturation, and its technique was simple and rapid. Results were obtained in 2.5 hours. By the analysis of the individual estrogen levels in urine collected every 4 hours from 20 IVF-ET patients, the following results were found, with a few exceptions: once the level had exceeded 60 ng/ml, the onset of the luteinizing hormone surge was induced within the next 72 hours; when the level had been under 60 ng/ml, the onset was not induced within the next 24 hours

In vitro fertilization was attempted during a spontaneous cycle in 17 out of 97 patients. Preovulatory urinary estrogen and LH maximas were assayed in order to time celioscopy 24 hours after the onset of the triggering urinary LH maxima. In 8 patients, comparison with plasmatic LH shows that the timing of spontaneous ovulation can be precisely predicted from urine assays. However, when the triggering LH release is weak, LH must be above or equal to 25 IU/g of creatinine with a creatininuria above or equal to 0.4 g/l in two urine samples at a three-hour interval.

Urinary oestrogen and pregnanediol excretion was measued daily ("daily monitoring") for a complete cycle in 20 normally menstruating women, in one patient with an anovulatory cycle and for 28 days in a patient with secondary amenorrhoea. The measurements were also performed on urine specimens collected at weekly intervals for 4 to 6 weeks ("weekly tracking") from 506 patients with evidence of abnormal ovarian function. These included 9 patients with primary amenorrhoea, 132 patients with secondary amenorrheoa, 138 patients with oligomenorrhoea and 227 patients with evidence of ovarian dysfunction and cycle lengths of 25 to 42 days. The results were subjected to statistical analysis. In the normal cycles, ovulation could be identified on the criteria of a rising pregnanediol value reaching or exceeding 2-0 mg. per 24 hours for a period of 7 days or more. Valid conclusions on the overall mean oestrogen and pregnanediol values for a complete cycle could be made from the results of weekly tracking, irrespective of which day the tracking commenced. Correlations were obtained by comparing the mean and maximum urinary oestrogen values and the variability of the values with the evidence of ovarian function indicated by the clinical classifications of the patients, the duration of the disorders and the subsequent occurrence of uterine bleeding. Mean oestrogen values of 10 mug. per 24 hours or less were associated with lack of ovarian function. For values higher than this a discriminant function based on both the mean oestrogen value and the variability of the oestrogen values was useful in predicting onset of spontaneous menstruation. A single urine specimen collected 4 to 8 days before onset of menstruation showing a raised pregnanediol value of 2-0 mg. per 24 hours or more provided a valid test for ovulation in women with regular cycles, and a single urine specimen giving an oestrogen value of 10 mug per 24 hours or less gave a valid indication of absent ovarian function in women with amenorrhoea for two years or more. In all other circumstances serial sampling at weekly intervals provided a valid assessment of ovarian activity. Application of these principles allows the greatest amount of information on ovarian function to be obtained with the greatest economy of effort.

Recent progress in the assay of urinary hormones has opened new opportunities for epidemiologists to study hormones and health outcomes. This is especially true for studies of female reproduction. The cyclic nature of female reproduction can be fully described only by continuous frequent measurements that, in order to be practical, require easily collected biological specimens. We describe our experience in collecting and analyzing daily urine specimens from 301 healthy women. We conclude that this approach is not only feasible but potentially of great value to epidemiologists for studying fertility, early pregnancy, the effects of toxic exposures on reproduction, and the relationships between reproduction and later risk of chronic diseases.

Immunoreactive estrogen and luteinizing hormone/chorionic gonadotropin (LH/CG) bioactivity have been measured throughout pregnancy in diverse primate species with a sample methodological approach utilizing small volumes of urine collected randomly. Total immunoreactive estrogen levels and the relative abundance of component estrogens (estrone, estradiol and estriol) at various stages of gestation were compared in the human, orangutan, pygmy chimpanzee, Douc langur and capuchin. Measurement of urinary LH/CG bioactivity enables an earlier detection of pregnancy when compared to the nonhuman primate pregnancy test. The methods described provide a practical and non-stressful way to detect and monitor pregnancy in widely divergent primate species.

The development of radioimmunoassay of steroid hormones in peripheral venous blood has led to a more direct index of gonadal production and concurrent monitoring of glandular function. While serial analyses of total urinary oestrogens can be correlated with luteinizing hormone (LH) excretion in treating amenorrhoea, radioimmunoassay of plasma oestrogens measures acute changes in ovarian responsiveness. For studies of androgens radioimmunoassay of plasma testosterone is the method of choice, but fractionation of urinary 17-oxosteroids may be useful and can be extended to include the pregnane series and corticoids. Urinary pregnanediol, however, may not reflect the endogenous secretion of progesterone and should be reserved for assessing induction of ovulation. Luteal function can be evaluated rapidly and specifically by radioimmunoassay of plasma progesterone. Although radioimmunoassay offers a new approach to ovarian physiology, its systematic application to disease is still in the future.

Estrogens and LH are necessary among other assays for ovulation diagnostic and ovarian monitoring during stimulations. They can be both measured with a method based on a bioluminescent reaction. This method measures, with a standard luminometer, the reduced NAD produced and accumulated by the reaction of two dehydrogenase enzymes: the estradiol dehydrogenase for direct assay of estrogens (estrone + estradiol), the glucose-6-phosphate dehydrogenase for gonadotrophin determination. These techniques can be applied to all biological fluids. They are simple and straightforward, do not need extraction and can be automated. Urinary assays are very useful in clinical practice to appreciate ovarian function, either during a spontaneous cycle or under stimulation, and are mandatory to decide the timing of hCG injection. The reported studies can be listed as follows: determination of preovulatory LH rise (LH greater than 10 Ul/g of creatinine) prior to embryo transfer after cryopreservation and thawing, detection of LH surges during ovarian stimulation, either premature surges causing premature luteinization, either normal surges when follicular maturation is adequate (296 cycles), confirmation of pituitary desensitization when using GnRH agonists (43 cycles), study of the initial stimulatory effect of GnRH agonists (13 patients). This effect can be responsible for the inadequate results obtained with the so-called "short protocol" in this experience when compared with the "long protocol" in the author's experience with compared with the "long protocol" (8 p. cent pregnancy rate per stimulation cycle versus 20 p. cent respectively, intrafollicular LH in 129 follicular fluids (86 with GnRH agonists and 43 without) has no correlation with the fecundability of the ovum. These results lead to extend bioluminescent techniques to the study of other parameters, and in particular FSH.

Twenty normally menstruating women volunteered for a study in which plasma samples were collected daily during an entire menstrual cycle. On the same days, samples of morning urine were also collected, as well as random samples of urine voided at the visit to the Outpatient Clinic. Progesterone (P), estradiol (E2) and lutropin (LH) were assayed in plasma, and pregnanediol-3-glucuronide (PdG), estrone-glucuronide (E1G), estriol-16-glucuronide (E3G), P, and E2 were measured in urine using radioimmunoassays. Progesterone in urine was assayed both with and without preceding chromatography. All urinary glucuronides and progesterone exhibited cyclic patterns similar to those of E2 or P in plasma. Seven-fold increases from early follicular to luteal phase values (for PdG and urinary P; the latter both with and without chromatography), or to peak levels (for E1G and E3G) were observed. The difference between the baseline and peak levels was less distinct (approximately 5-fold) for E2 in urine. The day-to-day coefficient of variation of early follicular phase values decreased from 40% to 25% by calculating the ratios of the glucuronides or P to creatinine (C). The peaks of estrogen glucuronides were delayed mostly by 1 day in comparison to the peaks of E2 in plasma. The urinary peaks of estrogens were in most cases more closely clustered around the day of the LH-peak when the measurements were corrected for C. For the determination of the first significant rise of steroid levels in a cycle, the calculation of a sustained rise (leading to a significant cumulative sum - CUSUM) was found superior when compared to other recommended indices, such as a 50% increase over the mean of 3 preceding values, or the increase over the baseline level plus 2 standard deviations. Sustained rises were calculated for all indices studied (including the ratio of urinary E1G to PdG). The ratio of E1G to C in morning urine gave consistently the most compact distribution of sustained rises. It is concluded that daily measurements of urinary PdG (or P) and E1G (or, possibly, E2) could substitute the serial assays of P and E2 in peripheral blood in the retrospective assessment of the ovarian functionn. The day-to-day variation can be significantly reduced, if results are expressed per concentration of C. For the prediction of ovulation or fertile period, the best index of urinary steroids appears to be the sustained rise in the ratio of E1G to C. However, this "best" method is still not good enough in terms of overall reliability and practicability.

Ten patients with oligomenorrhoea were investigated using increasing doses of clomiphene citrate in each cycle. The starting dose was 50 mg on day 1 of the cycle for 5 days and was increased by 50 mg per day for 5 days per cycle, to a maximum dose of 200 mg per day. Urinary total oestrogen concentration was measured on Day 1 to 3, 12 to 14 and on Day 21. Peripheral venous plasma concentrations of immunoreactive LH & FSH on Days 1 to 3 and 12 to 14 and the peripheral venous plasma concentration of immunoreactive progesterone on Day 21 were measured. The response to treatment is discussed in relation to hormone levels in the early follicular phase and to the dose of clomiphene administered. No correlation was found between hormone levels in the early follicular phase and increments in oestrogen and LH concentrations at midcycle. On a low dose regime the increment in FSH at midcycle was significantly greater in patients with a normal FSH concentration in the early follicular phase than in those with a low FSH level at this phase in the cycle. The results of treatment with clomiphene were not changed by an increase in dose from 50 mg to 150 mg but an increase to 200 mg significantly reduced the magnitude of response.

A practical approach for monitoring urinary estrogen and luteinizing hormone (LH) in various primate species is described. Total immunoreactive estrogen (Et) levels, measured without chromatography, provided a convenient estimation of estrone, estradiol and estriol as the sum of their separated parts. Bioactive LH in urine was measured by the rat interstitial cell testosterone (RICT) in vitro bioassay. Correction for urinary creatinine content allowed for the comparison of hormone production from small volumes of urine collected from single daily voidings. Using these methods, urinary Et and LH profiles through the reproductive cycles of a human, gorilla, chimpanzee and capuchin monkey have been determined.

The urinary gonadotrophin and ovarian steroid excretion pattern was studied in five women taking an oral contraceptive formulation consisting of mestranol 50 mug and norethisterone 1 mg. Both the pretreatment and post-treatment cycles were normal. The ovulatory peak of luteinizing hormone (LH) during the treatment cycles was uniformly suppressed, but LH continued to be excreted within the normal range. In one fifth of the treatment cycles there was a pronounced and sustained rise of oestrogen output in the absence of ovulation, and in many of the other treatment cycles oestrogen levels suggested that active ovarian steroidogenesis was taking place.

A practical approach for monitoring the reproductive cycle of primates is described. Ovulation, implantation, gestation and post-partum amenorrhoea were identified from measurements of total immunoreactive oestrogen and LH bioactivity in incomplete daily urine samples collected from various species, including a gorilla, a chimpanzee and an orang-utan. Oestrogen values were determined by radioimmunoassay of hydrolysed urine using a non-specific oestriol antiserum and LH was assessed by measuring testosterone output from dispersed rat Leydig cells. Both hormone measurements were indexed by urinary creatinine levels to help adjust for differences in urine concentration and volume.

Three women recorded their daily basal body temperature and collected 24-h urine specimens for total oestrogens and LH measurement during 12 consecutive cycles and one women did so for six consecutive menstrual cycles. The variability in the time relationship between these indices of ovulatory events in serial cycles is presented. In the pooled data the duration of the premenstrual phase from oestrogen peak, LH peak, and temperature rise, to the end of the cycle is 12.9, 12.1, and 10.8 days, respectively. The time differences between these indices are found to be biological constants.

The purpose of the present study was to find out whether there is a correlation between the urinary levels of testosterone or of estrogens (estrone and estradiol) and ICSH urinary concentration in normal postmenopausal women. Examination of 35 normal subjects at different postmenopausal ages (from one year to over 10 years after menopause) showed a highly significant inverse correlation between ICSH and testosterone urinary excretion levels (coefficient of correlation "r" equals minus 0.548), but none between the urinary estrogen excretion levels and ICSH urinary concentrations.

The preovulatory LH surge induces final maturation of the ovocyte which makes it suitable for in vitro fertilization. The spontaneous LH peak can be detected in blood or in the urine. Whatever the method of determination, the diagnosis of the triggering LH surge can be done only on strict criteria. This work defines these criteria when urinary Higonavis is used: a rise in the plus signs in two successive samples or a positive result in urine diluted fourfold.

The effect of 0.35 mg of norethisterone daily on cervical mucus and on urinary LH, pregnanediol and oestrogen levels was studied during two treatment cycles in five subjects who had been observed during a control cycle. Variable changes in hormone excretion patterns were found and are discussed.

Changes in the excretion of immunoreactive oestrogens and biologically active LH were assessed from measurements on small samples of urine collected from a capuchin, a tamarin, a spider monkey and a squirrel monkey. The hormone profiles were used to time matings of the capuchin and spider monkey; conception and pregnancy ensued. Detailed analysis of one urine sample from each female by using partition chromatography and reverse-phase high-pressure liquid chromatography showed that oestrone was more abundant than oestradiol-17 beta and oestriol combined, but the relative contribution of individual oestrogens to the total oestrogen complement differed. In the sample from the capuchin, an immunoreactive oestrogen slightly less polar than oestradiol appeared to be the most abundant urinary oestrogen.

Once weekly observations of the excretion of FSH, LH, oestrogens and pregnanediol have been used to monitor the changes which occur as New Zealand women approach and pass through the menopause. There were 3 patterns of hormone excretion. (1) Premenopausal women (aged 40-51 yr) had regular menstrual cyclicity with hormone patterns similar to those seen in the ovulatory cycles of fertile young women. (2) Women in the menopausal transition (40-55 yr) had irregular menstrual cyclicity with erratic hormone fluctuations. There were ovulatory cycles, postmenopausal episodes in which amenorrhoea was associated with high gonadotrophin levels and low urinary oestrogens, and times when the excretion of both gonadotrophins and oestrogens soared. Ovarian activity did not cease at the menopause, and postmenopausal women in the 6 months following final menstruation (44-55 yr) had hormone patterns which were indistinguishable from those observed in the long anovulatory cycles of the menopausal transition. (3) Older women (57-67 yr) had senescent ovaries with the unvarying high gonadotrophin and low oestrogen levels which are a consequence of ovarian failure.