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Urinary progesterone and pregnanediol: Use for monitoring progesterone treatment
Abstract
To compare progesterone (P) and pregnanediol glucuronide (PDG) levels in urine with respect to their potential use for monitoring luteal activity and P treatment.
Two different experiments were carried out. In the first experiment, each of seven normal, ovulatory women collected first morning urines daily throughout an entire menstrual cycle. The day of ovulation was determined by transvaginal ultrasound scanning. P, PDG, estrone glucuronide, luteinizing hormone and creatinine were measured in each urine specimen. In the second experiment, each of three normal, ovulatory women was given a single oral dose of 200 mg of micronized P and, 2 days later, a single intramuscular injection of 25 mg of P during days 2-5 of the cycle. Blood and urine were collected prior to each treatment and 1, 4, 8, 12 and 24 hours after treatment. P was measured in both serum and urine; PDG was quantified only in urine.
The mean initial rises and peak days, as well as the patterns of urinary excretion of P and PDG during the menstrual cycles, were similar; however, the variability of PDG was much greater. Concentrations of PDG were 1,000-4,000 times greater than those of P. A significant correlation was observed between urinary P and PDG. Following either intramuscular or oral P treatment, serum P levels rose rapidly and reached peak levels (7.0-11.8 ng/mL) by one hour. In contrast, both urinary P and PDG peaked considerably later (4-12 hours). Twenty-four hours after intramuscular treatment, serum and urinary P and urinary PDG were still elevated. Following oral treatment the levels of these compounds decreased after peaking and were 10- to 20- fold lower at 24 hours. Also, after oral P treatment, urinary PDG levels were considerably higher and more variable as compared to urinary P levels.
Urinary P appears to be as good a clinical marker of luteal activity and therapeutic P administration as PDG and may be measured in place of this metabolite. There appears to be greater variability in urinary PDG as compared to urinary P when P is administered.
... Serial transvaginal ultrasound (the gold standard) or blood sampling would not have been feasible, while menstrual diaries alone are not sufficiently accurate . Pregnanediol glucuronide (PDG), the major metabolite of P4, is 3,000 to 5,000 times more abundant than P4 in urine and assessed by manual ELISA (Stanczyk et al., 1997). We anticipated over 4,500 urine samples from the FEAT Study, which would be onerous to complete by manual ELISA. ...
... urine (Stanczyk et al., 1997). In practice, PDG assays often require a relatively onerous manual competitive ELISA, and the related expense limits the availability of testing in a clinical setting. ...
... The Architect method demonstrated a greater luteal rise for P4 than reported by Stanczyk et al. (1997), however this was still significantly less than was seen for PDG. A specific ELISA for PDG remains superior in confirming ovulation using urine samples to either automated P4 assay. ...
... Such elution was developed in order to apply this method for separation of other steroid-related hormones in untargeted metabolomics profiling approach. Taking into account the structure of steroid hormones and their derivates possibly detected in urine samples, the gradient elution set to 80% of B seems to be enough to chromatographically separate other steroid-related compounds (Stanczyk et al., 1997;Boyaci et al., 2016). ...
... The reason of that decreased level can be likely associated with another pathways of enzymatic biotransformation of these compounds. Progesterone can be metabolized to its main metabolite pregnanediol-3-glucuronide (PDG) (Stanczyk et al., 1997) so the balance between progesterone level itself can be moved to formation of other metabolites after enzymatic deconjugation like pregnanediol. Above all, the application of enzymatic hydrolysis step before the TF-SPME approach can be utilized in order to ensure better metabolome coverage in other untargeted metabolomics profiling studies. ...
In the present study, the development and optimization of a thin film solid phase microextraction method (TF-SPME) was conducted for metabolomics profiling of eight steroid compounds (androsterone, dihydrotestosterone, dihydroepiandrosterone, estradiol, hydroxyprogesterone, pregnenolone, progesterone and testosterone) from urine samples. For optimization of extraction method, two extraction sorbents (PAN-C18 and PS-DVB) were used as they are known to be effective for isolation of low-polarity analytes. The stages of sample extraction and analyte desorption were considered as the most crucial steps in the process. Regarding the selection of the most suitable desorption solution, six different mixtures were analyzed. As a result, the mixture of ACN: MeOH (1:1, v/v) was chosen in terms of the highest analytes’ abundances that were achieved using the chosen solvent. Besides other factors were examined such as the volume of desorption solvent and the time of both extraction and desorption processes. The analytical determination was carried out using the ultra-high performance liquid chromatography coupled with high resolution tandem mass spectrometry detection in electrospray ionization and positive polarity in a scan mode (UHPLC-ESI-QTOF/MS). The developed and optimized TF-SPME method was validated in terms of such parameters as extraction efficiency, recovery as well as matrix effect. As a result, the extraction efficiency and recovery were in a range from 79.3% to 99.2% and from 88.8% to 111.8%, respectively. Matrix effect, calculated as coefficient of variation was less than 15% and was in a range from 1.4% to 11.1%. The values of both validation parameters (recovery and matrix effect) were acceptable in terms of EMA criteria. The proposed TF-SPME method was used successfully for isolation of steroids hormones from pooled urine samples before and after enzymatic hydrolysis of analytes.
... From a sensor performance perspective, the LOD is 55 nM and the device is suitable for clinically relevant sample analysis. 59 Such a QD-based FRET sensor is also advantageous as it enables repeated analyte measurements. By contrast, immunoassays are considered effectively irreversible, stemming from fixation of antibody−analyte binding. ...
Immobilization of biosensors in or on a functional material is critical for subsequent device development and translation to wearable technology. Here we present the development and assessment of an immobilized quantum dot - transcription factor - nucleic acid complex for progesterone detection as a first step toward such device integration. The sensor is composed of a polyhistidine-tagged transcription factor linked to a quantum dot and a fluorophore-modified cognate DNA, and embedded within a hydrogel as an immobilization matrix. The hydrogel is optically transparent, soft, and flexible as well as traps the quantum dot - transcription factor DNA assembly but allows free passage of the analyte, progesterone. Upon progesterone exposure, DNA dissociates from the quantum dot - transcription factor DNA assembly resulting in an attenuated ratiometric fluorescent output via Förster resonance energy transfer. The sensor performs in a dose-dependent manner with a limit of detection of 55 nM. Repeat analyte measurements are also similarly successful. Our approach combines a systematically characterized hydrogel as an immobilization matrix and a transcription factor - DNA binding as a recognition/ transduction element, offering a promising framework for future biosensor devices based upon allosteric transcription factor.
... The detection of surges in luteinizing hormone (LH) and the rise in estrogen prior to the LH surge predict but do not verify ovulation 42 . Detecting progesterone surges confirms ovulation but typically requires blood testing 34,43,44 . Two at-home urine tests are currently being marketed 43,45 to measure pregnanediol-glucuronide. ...
Bacteria are an enormous and largely untapped reservoir of biosensing proteins. We describe an approach to identify and isolate bacterial allosteric transcription factors (aTFs) that recognize a target analyte and to develop these TFs into biosensor devices. Our approach utilizes a combination of genomic screens and functional assays to identify and isolate biosensing TFs, and a quantum-dot Förster Resonance Energy Transfer (FRET) strategy for transducing analyte recognition into real-time quantitative measurements. We use this approach to identify a progesterone-sensing bacterial aTF and to develop this TF into an optical sensor for progesterone. The sensor detects progesterone in artificial urine with sufficient sensitivity and specificity for clinical use, while being compatible with an inexpensive and portable electronic reader for point-of-care applications. Our results provide proof-of-concept for a paradigm of microbially-derived biosensors adaptable to inexpensive, real-time sensor devices. Bacteria represent an unexploited reservoir of biosensing proteins. Here the authors use genomic screens and functional assays to isolate a progesterone sensing allosteric transcription factor and use a FRET-based method to develop an optical progesterone sensor.
... The Architect method demonstrated a greater luteal rise for P4 than reported by Stanczyk et al. 12 , however this was still significantly less than was seen for PDG. A specific ELISA for PDG remains superior in confirming ovulation using urine samples to either automated P4 assay. ...
Urinary concentrations of the major progesterone (P4) metabolite pregnanediol-3-glucuronide (PDG) are used to confirm ovulation. We aimed to determine whether automated immunoassay of urinary P4 was as efficacious as PDG to confirm ovulation. Daily urine samples from 20 cycles in 14 healthy women in whom ovulation was dated by ultrasound, and serial weekly samples from 21 women in whom ovulation was unknown were analysed. Daily samples were assayed by two automated P4 immunoassays (Roche Cobas and Abbott Architect) and PDG ELISA. Serial samples were assayed for P4 by Architect and PDG by ELISA. In women with detailed monitoring of ovulation, median (95% CI) luteal phase increase was greatest for PDG, 427% (261–661), 278% (187–354) for P4 Architect and least for P4 Cobas, 146% (130–191), p < 0.0001. Cobas P4 also showed marked inaccuracy in serial dilution. Similar ROC AUCs were observed for individual threshold values and two-sample percent rise analyses for P4 Architect and PDG (both >0.92). In serial samples classified as (an)ovulatory by PDG, P4 Architect gave ROC AUC 0.95 (95% CI 0.89 to 1.01), with sensitivity and specificity for confirmation of ovulation of 0.90 and 0.91 at a cutoff of 1.67 μmol/mol. Automated P4 may potentially be as efficacious as PDG ELISA but research from a range of clinical settings is required.
... Ovarian hormonal activity can be accurately monitored with early morning urine samples (5,6). Serum LH, estradiol and progesterone correlate very well with urinary LH (7)(8)(9), estrone glucuronide (EG), and PG [ (10)(11)(12)(13)(14)]. Lately serum LH, estradiol and progesterone have been confirmed to correspond well with urinary LH, EG, and PG (15). ...
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.
... Though urine levels of progesterone have not been reported previously in any cetacean, the progesterone metabolite, pregnanediol-3 alpha-glucuronide (PDG), has been measured in several odontocete species (Walker et al. 1988, Kjeld 2001, Robeck et al. 2004). There appears to be a strong relationship between urine progesterone and PDG, suggesting a relatively simple, though crude, transformation that links the 2 values (Stanczyk et al. 1997, Falk et al. 1999. The urine progesterone values were similar to other mammals (Herrick et al. 2000, Khan et al. 2008) and comparable to the transformed PDG values reported in other cetaceans (Robeck et al. 2004(Robeck et al. , 2005. ...
ABSTRACT: Bowhead whale Balaena mysticetus progesterone concentrations were measured in different sample matrices (serum, blubber, and urine) to investigate (1) concordance among sample type and (2) variation among life-history class. Samples were collected from subsistence-hunted whales (n = 86) taken from 1999 to 2009. In general, irrespective of sample matrix, pregnant females had the highest concentrations by orders of magnitude, followed by mature animals of both sexes, and subadults had the lowest concentrations. Subadult males and females had similar progesterone concentrations in all sample matrices measured. When pregnant animals were included in our analyses, permuted regression models indicated a strong positive relationship between serum and blubber progesterone levels (r<sup>2</sup> = 0.894, p = 0.0002). When pregnant animals were not included, we found no significant relationship between serum and blubber levels (r<sup>2</sup> = 0.025, p = 0.224). These results suggest that progesterone concentrations are mirrored in these sample types over longer periods (i.e. on the order of weeks to months, time frame of reproductive changes) but not shorter periods (i.e. on the order of hours to days, time frame of daily fluctuations). This conclusion is consistent even for progesterone concentrations measured in females that had recently changed pregnancy states (either new mothers or newly pregnant animals), for which blubber progesterone levels seem to lag those in the serum. Finally, urine progesterone had statistically significant positive relationships with serum (r<sup>2</sup> = 0.136, p = 0.0460) and blubber progesterone (r<sup>2</sup> = 0.150, p = 0.0421). Our results suggest that progesterone concentrations first peak in the serum, then in the urine, and finally in the blubber.
Mobile point-of-care diagnostics are paramount for the provision of healthcare. Hormonal diagnostics are powerful tools to monitor timely changes in human physiology. Hormone concentrations in serum directly correlate with urine excretions with minor time delays. Therefore, rapid tests for hormones in urine have been widely used for decades as means of early diagnostics, particularly in lateral flow immunoassay formats. However, the challenge of reading and interpreting these binary tests remains. Here we present a method for utilizing mobile technologies to quantitatively read and interpret hormonal test strips. The method demonstrates the detection of a urinary by-product of progesterone, pregnanediol glucuronide (PdG), and its relation to ovulation and the fertility cycle.
Large intersubject variability is found in serum or plasma levels and pharmacokinetic parameters, including bioavailability and half-life, of all the progestogens. Higher bioavailability of a progestogen may result in a lower required dosage and less intersubject and intrasubject variability. A long serum half-life may be associated with a greater protective effect in the event of missed pills and may provide more consistent cycle control. Assessment of progestogen potency is problematic due to the large number of variables and assumptions associated with quantitative and qualitative tests of potency. Difficulties also arise when potency estimates from animal tests are extrapolated to humans. Nevertheless, certain clinical tests in which potency of orally administered progestogens is assessed in women can provide useful information from which optimal therapeutic progestogen doses can be determined. Biologic activities of progestogens, other than progestational effects, have been poorly studied. There is a misconception about progestin androgenicity. This is due primarily to extrapolation of data from rat studies to the human and misinterpretation of data that show effects of progestins on sex hormone-binding globulin and free testosterone. A better understanding of the pharmacokinetics and potency of progestins will help clinicians to choose the optimal type and dose of progestogen for individualized treatment of patients.
As discussed in previous papers at this symposium, steroids or other low-molecular-weight compounds are not inherently antigenic (Goodfriend and Sehon, 1970; and Gross, 1970). Landsteiner, however, demonstrated in the early 1900’s that antibodies directed against these low-molecular-weight compounds can be produced if they are chemically conjugated to a substance which is antigenic (see Landsteiner, 1946, for review). When conjugated to the antigen, these low-molecular-weight substances function as haptens.
Urinary excretions of progesterone (P) during normal menstrual cycle have been measured by radioimmunoassay. The excretion pattern mimics that of plasma P. Measurements of P in a single voided urine may be a reliable substitute for plasma analyses in the evaluation of ovulation and corpus luteal function.