Algorithms with Area under the Curve for Daily Urinary Estrone-3-Glucuronide and Pregnanediol-3-Glucuronide to Signal the Transition to the Luteal Phase.

Background and Objectives: Home fertility assessment methods (FAMs) for natural family planning (NFP) have technically evolved with the objective metrics of urinary luteinizing hormone (LH), estrone-3-glucuronide (E3G) and pregnanediol-3-glucuronide (PDG). Practical and reliable algorithms for timing the phase of cycle based upon E3G and PDG levels are mostly unpublished and still lacking. Materials and Methods: A novel formulation to signal the transition to the luteal phase was discovered, tested, and developed with a data set of daily E3G and PDG levels from 25 women, 78 cycles, indexed to putative ovulation (day after the urinary LH surge), Day 0. The algorithm is based upon a daily relative progressive change in the ratio, E3G-AUC/PDG-AUC, where E3G-AUC and PDG-AUC are the area under the curve for E3G and PDG, respectively. To improve accuracy the algorithm incorporated a three-fold cycle-specific increase of PDG. Results: An extended negative change in E3G-AUC/PDG-AUC of at least nine consecutive days provided a strong signal for timing the luteal phase. The algorithm correctly identified the luteal transition interval in 78/78 cycles and predicted the start day of the safe period as: Day + 2 in 10/78 cycles, Day + 3 in 21/78 cycles, Day + 4 in 28/78 cycles, Day + 5 in 15/78 cycles, and Day + 6 in 4/78 cycles. The mean number of safe luteal days with this algorithm was 10.3 ± 1.3 (SD). Conclusions: An algorithm based upon the ratio of the area under the curve for daily E3G and PDG levels along with a relative PDG increase offers another approach to time the phase of cycle. This may have applications for NFP/FAMs and clinical evaluation of ovarian function.

The Fertility Indicator Equation Using Serum Progesterone and Urinary Pregnanediol-3-Glucuronide for Assessment of Ovulatory to Luteal Phase Transition.

Background and Objectives: The Fertility Indicator Equation (FIE) has been shown to signal the fertile phase during the ovulatory menstrual cycle. It was hypothesized that this formulation, a product of two sequential normalized changes with a sign indicating direction of change, could be used to identify the transition from ovulatory to luteal phase with daily serum progesterone (P) and urinary pregnanediol-3-glucuronide (PDG) levels. Materials and Methods: Day-specific serum P levels from two different laboratories and day-specific urinary PDG levels from an additional two different laboratories were submitted for FIE analysis. These day-specific levels included mean or median, 5th, 10th, 90th and 95th percentile data. They were indexed to the day of ovulation, day 0, by ultrasonography, serum or urinary luteinizing hormone (LH). Results: All data sets showed a clear "cluster"-a periovulatory sequence of positive FIE values with a maximum. All clusters of +FIE signaled the transition from the ovulatory to luteal phase and were at least four days in length. The start day for the serum P and urinary PDG FIE clusters ranged from -3 to -1 and -3 to +2, respectively. The end day for serum P and PDG clusters went from +2 to +7 and +4 to +8, respectively. Outside these periovulatory FIE-P and FIE-PDG clusters, there were no consecutive positive FIE values. In addition, the maximum FIE-P and FIE-PDG values throughout the entire cycles were found in the clusters. Conclusions: FIE analysis with either daily serum P or urinary PDG levels provided a distinctive signature to recognize the periovulatory interval. The Fertility Indicator Equation served to robustly signal the transition from the ovulatory phase to the luteal phase. This may have applications in natural family planning especially with the recent emergence of home PDG tests.

Towards the Clinical Evaluation of the Luteal Phase in Fertile Women: A Preliminary Study of Normative Urinary Hormone Profiles.

Objective: To describe and evaluate urinary hormone profiles in the luteal phase. Setting and Patients: Twenty-five healthy fertile women, with regular ovulatory pattern cycles as assessed by temperature and cervical mucus, at a university based center. Methods: Daily urinary hormonal assessment of luteinizing hormone, estrone glucuronide, and pregnanediol glucuronide. This was done during 3 or more cycles, with 78 completed cycles. Samples were analyzed by both crude levels and levels adjusted for the hormone excretion rate. Correlation between measured parameters (LH surge, vulvar mucus) was assessed with regard to their ability to detect presumed ovulation. Results: An upper, middle, and lower tercile range for the main urinary reproductive hormones was determined and a classification system of zones proposed, considering profiles over or under the 10th percentile. Adjustment for the urine excretion rate proved useful for interpreting individual samples; this was less necessary with multiple samples over time where trends could be determined. This serial evaluation, in at least two cycles, lowered the possibility of finding an isolated luteal phase defect and helped identify the recurrence of such. Vulvar mucus findings performed well in determining the timing of ovulation. Despite the proven fertility of the study population, lower luteal phase hormones were detected in both an isolated and, in some situations, recurrent manner. Conclusion: A feasible method is proposed to accurately, thoroughly and reproducibly study the luteal phase in order to evaluate and treat identified abnormalities in a properly timed, restorative manner. This preliminary study provides the basis for future research, correlating urinary hormones with clinical findings, particularly those of luteal phase defects.

Monitoring of ovarian activity by measurement of urinary excretion rates using the Ovarian Monitor, Part IV: the relationship of the pregnanediol glucuronide threshold to basal body temperature and cervical mucus as markers for the beginning of the post-ovulatory infertile period.

STUDY QUESTION: Do the basal body temperature (BBT) shift and the cervical mucus markers for the beginning of the post-ovulatory infertile phase (POIP) of a menstrual cycle agree with the corresponding urinary pregnanediol glucuronide (PdG) threshold value? SUMMARY ANSWER: Perfect agreement between the cervical mucus markers and BBT shift and the hormonal definition of the start of post-ovulatory infertility occurred for only 7-17% of the cycles. WHAT IS KNOWN ALREADY: The PdG threshold of 7.0 µmol/24 h is an objective and accurate marker for the beginning of the POIP. The rise in serum progesterone also produces the BBT shift and changes in cervical mucus which determine the mucus peak. Serum progesterone and urinary PdG are closely correlated when variations in urine volume are taken into account. STUDY DESIGN, SIZE, DURATION: Individual menstrual cycle profiles of urinary PdG excretion rates for 91 fertile cycles from normally cycling women were analysed to identify the day of the beginning of the POIP. These days were compared with those determined by the day of the BBT shift +2 days, the day of the mucus peak +4 days and the later of these two indicators. The study lasted 3 years. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 62 women with normal menstrual cycles were recruited from three centres: Palmerston North, New Zealand; Sydney, Australia and Santiago, Chile. The cycles were displayed individually in a proprietary database program which recorded the PdG excretion rates, the BBT shift day and the cervical mucus peak day. A group of 15 women from a separate Chilean study had PdG urinary data measured as well as their day of ovulation determined by ultrasound. MAIN RESULTS AND THE ROLE OF CHANCE: The BBT and cervical mucus markers differed significantly in their identification of the beginning of the POIP when compared with the PdG excretion rate of 7.0 µmol/24 h. The observation that the BBT shift day and the mucus peak day could be identified even though the PdG excretion rates were still at baseline levels in some cycles could lead to an unexpected pregnancy for women using these natural family planning (NFP) indicators. LIMITATIONS, REASONS FOR CAUTION: The study consisted only of fertile cycles from women with regular cycles of 20-40 days duration. All the women were intending to avoid a pregnancy during the study, thus the limits of the fertile window were not tested. WIDER IMPLICATIONS OF THE FINDINGS: The NFP signals occurring earlier than the PdG threshold day could lead to an unexpected pregnancy. The signals occurring on the same day or later than the PdG threshold would not lead to unexpected pregnancies, but would require extra abstinence that could lead to non-compliance with the NFP method. A possible improvement in reliability of NFP methods is suggested. STUDY FUNDING/COMPETING INTERESTS: This study (project #90905) was funded by the NDP/UNFPA/UNICEF/WHO/World Bank Special Programme of Research, Development and Research Training in Human Reproduction (HRP). D.G.C. currently works for a diagnostic development company, Science Haven Ltd. The other authors have nothing to declare.

Cervicovaginal fluid changes to detect ovulation accurately.

OBJECTIVE: The purpose of this study was to evaluate changes in cervicovaginal fluid characteristics to identify ovulation. STUDY DESIGN: Several ovulation indicators were studied in a university-based natural family planning center. Fifteen parous women during 29 ovulatory cycles detected cervicovaginal fluid at the vulva. They self-aspirated their upper vaginal fluid, described it, and kept it for later checking. They also took basal body temperature, collected timed first morning urine samples for estrone and pregnanediol glucuronide enzyme immunoassays, and submitted to serial ovarian transvaginal ultrasound scans. RESULTS: Considering a +/-1-day period since ultrasound ovulation detection or allowing an extra day (-1 to +2), women perceived ovulation from cervicovaginal fluid at the vulva in 76% or 97% of cycles, on the basis of their visual description of vaginally extracted fluid in 76% or 90%, which rose to 90% or 97% for the instructor's description, and in 76% or 86% with a rapid drop in glucuronide ratio. Basal body temperature was less precise (71% or 79%). CONCLUSION: Evaluation of cervicovaginal fluid changes is an accurate ovulation indicator.

Mean versus individual hormonal profiles in the menstrual cycle.

OBJECTIVE: To evaluate hormonal profiles of normal menstrual cycles. DESIGN: Prospective, descriptive study of a case series. SETTING: University-based natural family planning center. PATIENT(S): Twenty-five natural family planning users for three or more cycles (n = 78). These women were healthy, contraception-free, parous, with regular ovulatory cycles. INTERVENTION(S): Immunoassays for estrone glucuronide, LH, and pregnanediol glucuronide were done in daily timed and measured samples of early morning urine. MAIN OUTCOME MEASURE(S): Estrone glucuronide, LH, and pregnanediol glucuronide levels were measured during the menstrual cycle. RESULT(S): All cycles showed an ovulatory pattern configuring classic hormonal mean curves. Most (77%) differed from the mean curve pattern. All had estrone glucuronide peaks, LH peaks, and pregnanediol glucuronide increases. Estrone glucuronide and LH peaks were not always clear; some lasted more than 1 day (long peak: estrone glucuronide 19%, LH 9%) or fluctuated (double peak: estrone glucuronide 4%, LH 6%; small LH peak: 19%). There were also prepeak estrone glucuronide surges, and pre- and postpeak LH surges. Pregnanediol glucuronide increased more clearly (6% fluctuated 1 day). Some women had repeated cycles with long estrone glucuronide peaks (16%) and fluctuations in LH surge (44%). CONCLUSION(S): Normal menstrual cycle hormonal profiles generally differ from mean curves, which are usually considered standard.