Using Quantitative Hormonal Fertility Monitors to Evaluate the Luteal Phase: Proof of Concept Case Study.

Several new quantitative fertility monitors are now available for at-home use that measure estrogen, luteinizing hormone (LH), and progesterone (PDG) in urine. This case report compares the Mira and Inito quantitative fertility monitors with the well-established qualitative ClearBlue fertility monitor. Three clinical scenarios were evaluated: a normal cycle, a prolonged luteinization cycle, and an anovulatory cycle. The identification of the luteal phase (or lack thereof in the case of anovulation) and the transition through the three processes of luteinization, progestation, and luteolysis were clearly demarcated with the help of quantitative LH and PDG. Quantitative fertility monitors have the potential to identify details of the luteal phase to help women with regular cycles and abnormal luteal phases to help target interventions for optimizing fertility.

Assessment of anovulation in eumenorrheic women: comparison of ovulation detection algorithms.

OBJECTIVE: To compare previously used algorithms to identify anovulatory menstrual cycles in women self-reporting regular menses. DESIGN: Prospective cohort study. SETTING: Western New York. PATIENT(S): Two hundred fifty-nine healthy, regularly menstruating women followed for one (n=9) or two (n=250) menstrual cycles (2005-2007). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Prevalence of sporadic anovulatory cycles identified using 11 previously defined algorithms that use E2, P, and LH concentrations. RESULT(S): Algorithms based on serum LH, E2, and P levels detected a prevalence of anovulation across the study period of 5.5%-12.8% (concordant classification for 91.7%-97.4% of cycles). The prevalence of anovulatory cycles varied from 3.4% to 18.6% using algorithms based on urinary LH alone or with the primary E2 metabolite, estrone-3-glucuronide, levels. CONCLUSION(S): The prevalence of anovulatory cycles among healthy women varied by algorithm. Mid-cycle LH surge urine-based algorithms used in over-the-counter fertility monitors tended to classify a higher proportion of anovulatory cycles compared with luteal-phase P serum-based algorithms. Our study demonstrates that algorithms based on the LH surge, or in conjunction with estrone-3-glucuronide, potentially estimate a higher percentage of anovulatory episodes. Addition of measurements of postovulatory serum P or urine pregnanediol may aid in detecting ovulation.

Bayesian analysis of risk factors for anovulation.

Two algorithms for assessing ovulatory status using daily urinary levels of oestrogen and progesterone metabolites have been applied to non-clinic-based, free-living populations of women. These relatively new methods for assessing ovarian function have been used to assess the potential adverse effects of occupational and environmental exposures, such as smoking, on the reproductive health of women. One algorithm has been validated against serum hormone measurements and gives good sensitivity and specificity for anovulation. However, a gold standard is generally not available in epidemiologic field studies in which these daily urine samples are collected. In this paper, we used Bayesian methods to estimate: (i) the probability of occurrence of anovulation, (ii) the sensitivity and specificity of the two algorithms, and (iii) the association between anovulation and smoking and other risk factors in the absence of a perfect test. We evaluated the two published algorithms for assessing ovulatory status, based on their cross-classified results applied to one randomly selected cycle from each woman in a sample of 338 employed women. We first assumed that the algorithms were independent, conditional on ovulatory status. Then, we used a dependence model to allow for correlation between the results of the two algorithms. We implemented a Bayesian logistic regression analysis that allowed the outcome measurement to be partially imperfect. We incorporated the posterior distributions for algorithm accuracy obtained from the dependence model as prior distributions for this logistic regression model. Then, we compared the results with those obtained from a standard multiple logistic approach using the algorithm determination of ovulatory status as if it were perfect. Our results indicated that increasing physical activity was associated with a significantly increased risk of anovulation; and smokers had a potentially, but not statistically significant, increased occurrence of anovulation.

Psychological stress in the workplace and menstrual function.

The relation between psychological stress at work and menstrual function was examined for 276 healthy, working, premenopausal women who participated in the California Women's Reproductive Health Study in 1990-1991. Subjects collected daily urine samples and completed a daily diary for an average of five menstrual cycles. Metabolites of estrogen and progesterone were measured in the urine, and computer algorithms were developed to characterize each cycle as ovulatory or anovulatory and to select a probable day of ovulation. A telephone interview collected information about psychological stress at work as well as other occupational, demographic, lifestyle, and environmental factors. Logistic regression was used to model stressful work and risk of anovulation (> or = 36 days without ovulating) and measures of within-woman cycle variability. Repeated measures analyses were performed on other menstrual cycle parameters. Stressful work (high demand in combination with low control) was not strongly related to an increased risk for anovulation or cycle variability or to any of the following cycle endpoints: short luteal phase (< or = 10 days), long follicular phase (> or = 24 days), long menses (> or = 8 days), or long cycle (> or = 36 days). However, women in stressful jobs had a more than doubled risk for short cycle length (< or = 24 days) compared with women not working in stressful jobs (adjusted odds ratio = 2.24, 95% confidence interval 1.09-4.59).

Urinary sex hormone excretions in premenopausal women and coronary heart disease risk: a nested case-referent study in the DOM-cohort.

The low incidence of coronary heart disease (CHD) in premenopausal women is partly ascribed to protection by endogenous estrogen production. As a consequence, we hypothesized that premenopausal women with low endogenous estrogen production or high androgen production might be at increased risk for CHD. We studied the relationship between urinary sex hormone excretions and CHD risk by means of a nested case-referent study within a cohort of premenopausal (ages 40-49 yrs) women (n = 11,284). This cohort was formed at a breast cancer screening project in 1982-1986 (The Diagnostisch Onderzoek Mammacarcinoom [DOM] Project). Baseline data included self-administered questionnaires and anthropometric measurements. At the time of screening the women were instructed to collect an overnight urine sample on day 22 of three separate cycles. These urine samples were stored at -20 degrees C. Up to June 1991, 45 subjects were admitted to local hospitals on diagnosis of CHD (29 with myocardial infarction, and 16 with angiographically confirmed coronary disease). Referents were sampled from the cohort, matched for age and year of screening in a 1:3 ratio. In a follow-up study, menopausal state of the subjects was assessed yearly by mailed questionnaires. Urinary excretions of estrone-glucuronide, pregnanediol-glucuronide, and testosterone-glucuronide adjusted by creatinine were similar for cases and referents. Cases had no earlier menopause than referents, although cases had more anovulatory cycles. The occurrence of CHD in middle-aged women is not preceded by a low premenopausal endogenous estrogen production or high androgen production. Anovulatory cycles appear more frequently in women who develop CHD many years later.

Identification of anovulation and transient luteal function using a urinary pregnanediol-3-glucuronide ratio algorithm.

The sensitivity and specificity of a urinary pregnanediol-3-glucuronide (PdG) ratio algorithm to identify anovulatory cycles was studied prospectively in two independent populations of women. Urinary hormone data from the first group was used to develop the algorithm, and data from the second group was used for its validation. PdG ratios were calculated by a cycles method in which daily PdG concentrations indexed by creatinine (CR) from cycle day 11 onward were divided by a baseline PdG (average PdG/Cr concentration for cycle days 6-10). In the interval method, daily PdG/CR concentrations from day 1 onward were divided by baseline PdG (lowest 5-day average of PdG/CR values throughout the collection period). Evaluation of the first study population (n = 6) resulted in cycles with PdG ratios > or = 3 for > or = 3 consecutive days being classified as ovulatory; otherwise they were anovulatory. The sensitivity and specificity of the PdG ratio algorithm to identify anovulatory cycles in the second population were 75% and 89.5%, respectively, for all cycles (n = 88); 50% and 88.3% for first cycles (n = 40) using the cycles method; 75% and 92.2%, respectively, for all cycles (n = 89); and 50% and 94.1% for first cycles (n = 40) using the interval method. The "gold standard" for anovulation was weekly serum samples < or = 2 ng/ml progesterone. The sensitivity values for all cycles and for the first cycle using both methods were underestimated because of apparent misclassification of cycles using serum progesterone due to infrequent blood collection. Blood collection more than once a week would have greatly improved the sensitivity and modestly improved the specificity of the algorithm. The PdG ratio algorithm provides an efficient approach for screening urine samples collected in epidemiologic studies of reproductive health in women.

Interrelation between Western type cancers and non-Western type cancers as regards their risk variation in time and space. IV. Hormonal transition of Japanese women from the pro-cervical cancer age through the pro-endometrial cancer age to the pro-hypogonadism age.

Chronological trend of urinary steroid excretions in Japanese women was investigated during the period of June 1972 to August 1986 using healthy women of urban and rural origins, patients with breast cancer and patients with either cervical cancer or endometrial cancer. The excretions of 14 neutral steroids were estimated by gas liquid chromatography, and the obtained data were tentatively correlated with the epidemiological backgrounds. In the course of the chronological transition from the 1st stage (1972-1974) to the 2nd stage (1975-79), the urinary steroid pattern of Japanese women with and without cancer experienced a common change to produce specific deviations that were in agreement with the hormonal characteristics of a pill user or of an endometrial cancer patient. At the 3rd stage (1980-86), patients with either cervical cancer or endometrial cancer were distinguished from 1st stage controls by non-specific depression of all androgens, progestins and corticosteroids in urine. Throughout the whole period, both the risk for cervical cancer and the reproductive activity (birth rate) were found to decrease continuously in Japanese women. Evidence was presented to suggest that the above deterioration of the hormonal environment in Japanese women could be related to the stress of modern life rather than to defects in the diet. On the basis of the above findings, the 1st, 2nd and 3rd stages of our investigation were tentatively termed the pro-cervical cancer age, the pro-endometrial cancer age and the pro-hypogonadism age. The relation between the chronological change of urinary steroids and that of the epidemiological background was analyzed from the view point of population ecology.

[Possibilities for using the LH color test in determining ovulation time]. / Einsatzmöglichkeiten des LH-Color-Tests zur Ovulationszeitbestimmung.

The LH color test was used to monitor 40 cycles of 32 patients. The urine test, which becomes positive at a LH-concentration of at least 50 I. U./ml, indicated the LH-peak in 34 out of 35 cycles with ascertained ovulation. In 24 cycles, the urine LH-peak preceded the serum LH-peak by one day. The urine LH-peak was found on cycle day 13.6 on average. The serum LH-peak on day 14.2 on average. The sensitivity of the LH color test was 97.2%, its specificity 100%.

Application of penicillinase linked ELISA of pregnanediol glucuronide for detection of ovulation and assessment of corpus luteal function.

A penicillinase linked enzyme immunoassay was developed for the estimation of pregnanediol-3 alpha-glucuronide (PdG) in urine. The immunoassay satisfied all the validity criteria and was used in detecting ovulation and in the assessment of corpus luteal function (CLF) during spontaneous or induced cycles. Reference values were established by estimating PdG levels in daily early morning urine samples during 31 menstrual cycles obtained from 17 regularly menstruating women. A PdG value of 1.7 micrograms/mg creatinine (micrograms/mgC) (90th Centile of follicular phase) in any MLP (mid-luteal phase) sample was considered as indicating ovulation. A value of 4.6 micrograms/mgC (20th centile of MLP) was considered to be evidence of sufficient CLF. When this approach was applied to 20 infertile cases, detection of the occurrence of ovulation/anovulation was made correctly in 19 out of 20 cases (95%). Accuracy was poor (55.6%) when the aim of the diagnosis was corpus luteal deficiency. Higher accuracy (88.9%) for corpus luteal deficiency/corpus luteal adequacy was obtained when the sum of PdG concentrations in three MLP samples were taken into consideration. A total of 13.8 micrograms/mgC (thrice the 20th centile for MLP) indicated probable corpus luteal deficiency, and values above this limit were considered to indicate corpus luteal adequacy.

Rapid immunochemical assay of pregnanediol-3-glucuronide in urine and its clinical application.

A rapid and simple technique for assessing ovulation and luteal function is needed in clinical practice. A sensitive immunoassay kit is presented here for clinical use. An immunoassay of pregnanediol-3-glucuronide (Pd-3G) in urine was established. The sensitivity of this assay was 0.4 mg/l of Pd-3G. The standard error range of the recovery test was within 0.8 mg/l, and the coefficients of variation of within-run and day-to-day precision test were within 4%. Cross reactions of various steroids were observed in free pregnanediol, pregnanetriol and progesterone, but their reactivity exhibited less than 4%. The Pd-3G/creatinine (mg/g) ratio was demonstrated as nearly constant over a 24-hour period. A significant rise of Pd-3G coincided with a luteinizing hormone surge and continued during the luteal phase in the ovulatory menstrual cycle. In an anovulatory cycle, Pd-3G was found to be under 4 mg/g creatinine. This Pd-3G direct-assay method could be widely used in infertility clinics.

[An immunochemical assay of pregnanediol 3-glucuronide in urine and its variation in the peri-ovulatory period].

A rapid, simple and sensitive immunochemical assay of pregnanediol 3-glucuronide (Pd-3G) in urine was established. The standard error range of the recovery test was within 0.8 microgram/ml, and the coefficient variation of the within run and day to day precision tests were within 4%. Cross reactivity of various steroids exhibited less than 4%. A significant rise of Pd-3G (mg)/creatinine (g) coincided with the LH surge and continued during the luteal phase in the ovulatory menstrual cycle. In an anovulatory cycle, Pd-3G was found to be under 4 mg/g. This Pd-3G direct assay method could be widely used for assessing ovulation and luteal function.

[The random urine estrogen, pregnanediol/creatinine ratio determinations for monitoring ovarian function].

Study was made to assess the potential clinical value of a random urine estrogen, pregnanediol/creatinine ratio (E/C, P/C) determinations as an index of monitoring ovarian function. Radioimmunoassay of total estrogens and pregnanediol was carried out on 24 hour and paired early morning urine samples in 14 healthy volunteers and 4 infertile patients treated at our clinic during 20 menstrual cycles. Estriol-16-glucuronide and pregnanediol-3-glucuronide antisera were used for radioimmunoassay of total estrogens and pregnanediol, and assays were made without any treatment of urine samples such as hydrolysis and/or extraction. E/C and P/C ratios correlated well with 24 hour excretion of these hormones: Total estrogens (micrograms/24 hr) = 0.89 spot E/C (ng/mg/ml) + 0.21, r = 0.924, p less than 0.01, pregnanediol (mg/24 hr) = 1.28 spot P/C (micrograms/mg/ml) -0.37, r = 0.791, p less than 0.01. The data indicate that ovarian activity in women could be predicted by measuring estrogen, pregnanediol to creatinine ratios in small samples of urine, and the collection of 24 hr samples is unnecessary. In view of this, the conventional basal body temperature method, used up to the present time as an infallible guide in monitoring ovarian function, needs to be reappraised.

Anovulation and increased androgenic activity as breast cancer risk in women with fibrocystic disease of the breast.

The urine of 26 otherwise healthy women with fibrocystic disease of the breast was assayed by gas chromatography for testosterone and androstanediol (5alpha-androstane-3alpha, 17beta-diol), the major metabolite of dihydrotestosterone. The mean values for both androgens were significantly higher than in 18 normal women in the same age range. Sixteen of the 26 fibrocystic disease patients also had endometrial hyperplasia. Since the endometrial specimen was obtained in the premenstrual period, the presence of hyperplasia proved that the menstrual cycle in over two-thirds of the fibrocystic disease patients was nonovulatory.

PIP: Urine of 26 otherwise healthy women with fibrocystic breast disease (FCD) was assayed by gas chromatography for testosterone (Tes) and androstanediol (Ans), the major metabolite of dihydrotestosterone. The 26 patients under study had been subjected 3-5 months previously to biopsies of breast lumps, diagnosed as FCD. The histological features varied from simple cystic formations with moderate epithelial proliferation in 11 women (Group 1) to pronounced intraductal epithelial hyperplasia accompanied by epithelial cell atypia in 15 women (Group 2). The urinary Tes and Ans difference in FCD patients and controls (18 normal women) was significant at the level of p .01 for both androgens. In controls the mean excretion levels of Tes and Ans were 6.5 and 35 mcg/24 hours, respectively. In FCD patients, the mean Tes and Ans values were 17.4 and 68.5 mcg/24 hours, respectively. Group 2 presented a higher urinary Tes level than patients in Group 1, but the difference was not significant. The Ans level of Group 1 patients was significantly above normal (p .01) and near significantly higher (p .08) than that of the Group 2 patients; whereas the Ans level of Group 2 patients did not differ significantly from the normal value. Endometrial specimens showed that 16/26 FCD patients had endometrial hyperplasia. Since the endometrial specimen was obtained in the premenstrual period (Days 20-22), the presence of hyperplasia proved that the menstrual cycle in over two-thirds of the FCD patients was nonovulatory.

[Uncommonly high concentration of estrone of adrenal origin in a case of androgenic obesity, anovulation and amenorrhea]. / Niezwykle wysokie wartosci estronu pochodzenia nadnerczowego w przypadku androgennej otylosci,braku owulacji i miesiaczki.

PIP: The case of a 35-year-old woman who demonstrated androgenic obesity, absence of ovulation, and amenorrhea is examined. This patient showed arterial hypertension, diabetes mellitus, hirsutism, and anovulatory cycles. A very high concentration of estrone was noted in the urine, originating in the adrenal glands. These indications are generally considered during evaluation of breast or uterine cancer threat. Administration of dexamethasone led to a decrease in urinary estrone to insignificant levels. Stimulation with human chorionic gonadotropin caused an increase in ovarian activity. The disruptions this patient suffered were attributed to hormonal imbalances attributed to her obesity, primarily in regard to estrogen metabolism.^ieng

Analysis of variables in treatment of anovulation with human gonadotrophins.

Twenty seven women with anovulation were given 121 courses of gonadotrophins monitored by excretion of estrogen and pregnanediol. The FSH, ovulating dosage of HCG, time interval between FSH and HCG and subsequent supporting doses of HCG were varied in a statistical design. None significantly affected ovulation or pregnancy rates. The preparation with an FSH:LH ratio of 1 required a lower dosage to initiate an estrogen response than that with a ratio of 5. After adjustment it still produced a faster rise in estrogen excretion. The amount of estrogen also varied with the time interval between FSH and HCG, the ovulating dose of HCG and supporting doses of HCG. The amount of pregnanediol varied with the ovulating dose of HCG and the length of the luteal phase varied with the ovulating dose of HCG and supporting injections of HCG. The dosage of FSH needed to initiate a response increased on average by 8% from one course to the next. Simplicity, economy and risk of multiple pregnancy are discussed in relation to these variables.