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PubMed1975

The investigation of ovarian function by measurement of urinary oestrogen and pregnanediol excretion.

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.

Amenorrhea↗
PubMed1991

Progesterone and 5 beta-pregnanediol production by isolated fetal placental binucleate cells from sheep and goats.

Enzymic dispersion and density gradient separation were used for the isolation of enriched populations (60-90%) of cells from the corpus luteum, placenta and peripheral blood of pregnant sheep and goats. Analysis of the steroids produced from radioactive pregnenolone demonstrated that placental binucleate cells can produce progesterone and 5 beta-pregnanediol whereas white blood cells were relatively inactive. Thus, sheep binucleate cells converted pregnenolone predominantly to progesterone as did sheep luteal cells. However, goat binucleate cells produced 5 beta-pregnanediol as the major metabolite, which is consistent with its production in vivo during pregnancy. Production of progesterone (sheep) or 5 beta-pregnanediol (goat) by binucleate cells was shown to be proportional to the number and viability of the cells. In contrast with the binucleate cells there was no evidence that trophectodermal uninucleate cells play a significant role in placental progesterone or 5 beta-pregnanediol synthesis in either species.

Animals↗
PubMed1997

Does creatinine adjustment of urinary pregnanediol glucuronide reduce or introduce measurement error?

Correlation between urinary pregnanediol and serum progesterone measurements and the influence of age, race, smoking and urinary creatinine adjustment was determined during the luteal phase of the menstrual cycle in 175 volunteers. A decline in serum progesterone was observed with increasing age. Mean baseline urinary creatinine declined with increasing age in non-smokers and was not affected by race or baseline weight. An excellent correlation between urinary pregnanediol glucuronide and serum progesterone levels existed except when urinary pregnanediol concentrations were adjusted using creatinine measurements in older individuals. Adjustment of urinary pregnanediol glucuronide concentration using creatinine measurement is therefore discouraged.

Adult↗
PubMed1975

A modification of the Detter and Klingmüller's method for urinary alpha- and beta-pregnanediol and pregnanetriol determination: excretion patterns in men and in women during the normal menstrual cycle.

This report describes the modification of Detter and Klingmüller's method for the determination of urinary alpha- and beta-pregnanediol and pregnanetriol by means of thin -layer chromatography. The modifications were as follows: 1. The addition of formol prior to hydrolysis to prevent pigments penetration into urinary extracts. 2. The use of Kieselgel HF 254+366 nach Stahl (E. Merck, Darmstadt) which allows to localize the spots under UV-light at 366 nm without the use of colour reagents. The results concerning accuracy, sensitivity, precision and specificity in the modification described are profitable. Using this method in 7 healthy women (aged 28-37) with normal menstrual cycles the urinary excretion of alpha- and beta-pregnanediol and pregnanetriol were evaluated every second day (starting the 6th day of the cycle). The maximum of excretion of alpha- and beta-pregnandiol appeared on day 20 of the cycle, with the mean (+/- S.D.) 1.27 +/- 0.37 mg/24hr and 2.97 +/- 0.80 mg/24hr for alpha- and beta-pregnanediol, respectively. Mean values of pregnantriol were at the same level and ranged from 0.25 to 1.42 mg/24 hr. Single determination of these compounds in 8 healthy men (aged 19-42) revealed the mean excretion values (+/- S.D.) 0.96 +/- 0.17 mg/24 hr, 1.24 +/- 0.40 mg/24 hr, 1.12 +/- 0.65 mg/24 hr for alpha- and beta-pregnanediol and pregnanetriol, respectively.

Adult↗
PubMed1986

Steroid spectrum in human urine as revealed by gas chromatography. VI. Pregnanediol excretion values and 3 beta-hydroxy-delta 5-steroid dehydrogenase activity of girls at different stage of development.

Pregnanediol excretion in the urine of girls aged 2-14 years was found to be 0.13 +/- 0.17 mg/day. Before puberty the amount of pregnanediol excreted was low but a marked increase could be observed during puberty. In the case of premenarchal girls this increase correlated well with increasing body weight, while in the presence of cycle ovarian function (anovulational cycles) it exceeded the expected level. The activity of 3 beta-hydroxy-delta 5-steroid dehydrogenase was assayed in vivo indirectly, by measuring pregnanediol (pregnanediol and androsterone + ethiocholanone) DHA ratios from urine. A composition of the two ratios indicated that the utilization of the endogenous C21 intermediate was one order of magnitude less than that of the C19 intermediate. To interprete this finding it is assumed that the delta 5-3 beta-HSD of the liver is on allotype of the adrenal enzyme with a reduced activity, towards the C21 intermediate. The biological significance of this presumption is also discussed.

3-Hydroxysteroid Dehydrogenases↗
PubMed1977

[Effect of spirolactones on the urinary excretion of individual 17-oxosteroids and of pregnanediol in man (author's transl)].

The effect of spirolactones on the urinary excretion of individual 17-oxosteroids and of pregnanediol in man was investigated under various conditions. After purification by thin-layer chromatography, the 17-oxosteroids and pregnanediol were determined by gas chromatography. Eleven male subjects (age 21-34 years) received a daily dose of 200 mg spironolactone (Aldactone) orally on seven consecutive days. Five female subjects (age 20-35 years) and ten pregnant women (age 20-35 years; 28th.-39th. week of gestation) were given a daily intravenous injection of 600 mg potassium canrenoate (Aldactone pro injectione) on three consecutive days. In the male subjects, the excretion of individual 17-oxosteroids and of pregnanediol was significantly reduced during administration of spironolactone. In the nonpregnant female subjects, the excretion of etiocholanolone and dehydroepiandrosterone was diminished, whereas in the pregnant women, the excretion of pregnanediol and dehydroepiandrosterone was decreased during treatment with potassium canrenoate. These results suggest that the reduced urinary excretion of steroids may be due to an inhibition of steroid metabolising enzymes by spirolactones. It seems likely that spirolactones affect the enzymatic conversion fo cholesterol to pregnenolone.

17-Ketosteroids↗
PubMed1990

Monoclonal antibodies to pregnanediol-3-glucuronide: application to a direct enzyme-linked immunosorbent assay of urine.

Monoclonal antibodies to pregnanediol-3-glucuronide were produced and characterized. One of three clones investigated provided antibody suitable for a direct urinary enzyme-linked immunosorbent assay (ELISA). The ELISA uses a pregnanediol-thyroglobulin conjugate adsorbed onto the wells of a standard 96-well microtiter plate. Pregnanediol-3-glucuronide in standards or diluted urine competes with the immobilized steroid for antibody-binding sites. After washing, mouse monoclonal antibody bound to the plate is probed with antimouse immunoglobulin peroxidase. After further washing, o-phenylenediamine substrate is added and, finally, the absorbance is read at 492 nm. The ELISA shows excellent performance and agreement with the previous gas chromatographic method. The ELISA is ideal for aiding the assessment of ovarian function in the routine laboratory.

Animals↗
PubMed1990

Comparison of endometrial biopsy and urinary pregnanediol glucuronide concentration in the diagnosis of luteal phase defect.

To determine if pregnanediol glucuronide (PG) excretion is useful in luteal phase assessment, we compared daily first morning urinary PG concentrations during the luteal phase in nine normal and nine deficient cycles. Total luteal pregnanediol excretion (44.1 +/- 11.3 versus 64.0 +/- 11.6 area units +/- SEM) was not different. However, significantly less pregnanediol was excreted by the abnormal group during the 1st 5 days of the luteal phase (12.7 +/- 1.2 versus 18.0 +/- 1.7 area units +/- SEM, respectively). Thus, delayed PG excretion may be characteristic of luteal phase defect and measurement of urinary PG may be useful only if daily samples during the early luteal phase are obtained.

Biopsy↗
PubMed1977

An improved method for the simultaneous determination of individual 17-oxosteroids and of pregnanediol in urine by gas-liquid chromatography.

An improved method for the simultaneous gas-chromatographic determination of individual 17-oxosteroids and of pregnanediol in the same sample of urine is described. The method involves the following steps. The urinary sample is chromatographed on Amberlite XAD-2. The fraction containing the steroid conjugates is evaporated; the residue is dissolved in n-heptane and the organic phase extracted with water. The steroid glucuronides are hydrolysed by a bacterial beta-glucuronidase; subsequently, the steroid sulphates are solvolysed in ethyl acetate. Free 17-oxosteroids and pregnanediol are separated by two-dimensional thin-layer chromatography. The zone containing the steroids to be investigated is eluted; after evaporation, the residue is treat with N-methyl N-trimethyl silyl trifluoroacetamide. The reaction mixture is injected into a gas chromatograph; the trimethyl silyl ethers of the individual 17-oxosteroids and of pregnanediol are quantitated by measuring the peak heights.

17-Ketosteroids↗
PubMed1980

The effect of conception on the concentration of urinary oestrone-3-glucuronide, LH/hCG and pregnanediol-3 alpha-glucuronide.

The concentrations of oestrone-3-glucuronide, LH/hCG and pregnanediol-3 alpha-glucuronide have been measured in daily samples of early morning urine from 6 women throughout the normal ovarian cycle and from 5 subjects, who conceived during the period of study. In addition, the ratio of oestrone to pregnanediol glucuronide was calculate as an additional index of ovarian function. The results showed that the ratio during the follicular phase of the cycle was similar in both groups of women. Moreover, there was no significant difference between the groups in either the duration or intensity of the LH peak. Seven days later, however, the women who conceived excreted significantly higher amounts of oestrone-3-glucuronide (P less than 0.05) and 12 days after the LH peak the values for pregnanediol-3 alpha-glucuronide and LH/hCG were also elevated (P less than 0.05). A defined increment, in the concentration of hCH (3 times the mean basal level) occurred 11 to 15 days after the peak of LH and appeared 3 to 5 days after the increased excretion of the steroid glucuronides.

Adult↗
PubMed1988

Pregnanediol excretion in fertile women: age-related changes.

In normal women reproductive capacity diminishes with age; the decline has been detected before the start of the menopausal transition. It is known that in premenopausal women most menstrual cycles are ovulatory. An investigation was set up to examine the possibility that there is an age-related decline in the ability of the corpus luteum to secrete progesterone at this time. Once-weekly urine samples for the measurement of pregnanediol were collected from 100 women aged 20-48 years, all of whom had regular 20- to 35-day menstrual cycles (1124 samples collected during the course of 312 menstrual cycles of which 96.8% were ovulatory). Pregnanediol excretion rates parallel the levels of progesterone in plasma. Examination of the rank correlation between age and pregnanediol excretion identified a significant negative correlation during the early and mid-follicular phases, but failed to detect any age-related change during the luteal phase. The evidence does not support the concept of an age-related increase in luteal phase defects before the start of the menopausal transition.

Adult↗
PubMed1975

Serotonin, 5-hiaa, total estrogen and pregnanediol excretion in urine during therapeutic saline abortion.

In 24 patients, who underwent therapeutic abortion for various reasons between the 17th and 26th week of pregnancy, urinary excretion of serotonin, 5-HIAA, total estrogens, and pregnanediol were measured before, during and after the intra-amniotic injection of hypertonic saline. 20% hypertonic saline solution (160-500 ml) was given by transabdominal injection over a period of 5 min. The four hormones or metabolites were measured during six periods: I; 12-24 hrs, and II: 0-12 hrs before saline administration, III: 0-12 hrs after saline administration, IV: 0-12 hrs during aborion, V: 0-12 hrs and VI: 12-24 hrs after abortion. The results point to the active participation of serotonin in the process of fetal expulsion, as serotonin was increased by over 100% (from 20-22 to 43-47 mu-g/12 hrs) during periods III-IV, and its metabolite 5-HIAA, too, increased by nearly 60% (from 2.4-2.5 to 3.3-3.9 mg/12 hrs). They decreased during the post-abortive periods V-VI. On the other hand, total estrogens decreased only slowly, but continuously, during all 6 periods (4.9, 4.3, 3.4, 3.1, 1.8 and 1.4 mg/12 hrs). Pregnanediol, beginning with 12 mg/12 hrs showed a slight increase during periods III-IV (14.5 and 15.6 mg/12 hrs) and a decrease during periods V-VI (8.0 and 5.8 mg/12 hrs). These findings are interpreted as indicating the disruption of feto-placental function affecting estrogens during periods III-VI. They might demonstrate an accelerated hydrogenation of progesterone into pregnanediol during periods III-IV, followed by a sharp decreased in progesterone/pregnanediol production during periods V-VI.

Abortion, Therapeutic↗
PubMed1976

[Use of an ion exchanger as a rapid means of extraction, applied to the determination of urinary estrogens and pregnanediol in pregnancy, using colorimetry or gas chromatography].

The method described permits, after classical hydrolysis, extraction without a solvent of estrogens during pregnancy. It also permits estimation of pregnanediol which is eluted in a different fraction. Twenty estimations may be carried out in half a day on test samples of from 2 to 10 ml of urine. The estimation of total estrogen by colorimetry takes into consideration the efficiency of extraction by the use of a standard range treated under the same conditions. For gas chromatography, two internal standards are added to the urine after hydrolysis: these are 16,17-epiestriol and 5 beta pregnane-3 alpha, 17 alpha, 20 beta triol (PGTS) have respectively the same behaviour as estriol on the one hand and pregnanediol and pregnanetriol, on the other hand. They take into consideration resin extraction and chromatographic behaviour and may be used for quantitative estimation. The graphs of standardisation and recovery have been studied for the main estrogens together with pregnanediol. The various eluates were controlled by gas chromatography coupled to mass spectrometry.

Chromatography, Gas↗
PubMedDate not supplied

[The urinary excretion of pregnanediol during pregnancy determined by gas-liquid chromatography. III. Its relation with the obstetric results (author's transl)].

269 determinations of urinary pregnanediol by gas-liquid chromatography were made in 140 patients during pregnancy. A relation between the pregnandioluria and the fetal results (adequacy of the weight to gestational age, Apgar score at one and five minutes of birth, umbilical artery blood PH and perinatal mortality) as well as the global evaluation of the placenta (macroscopic aspect, vascular index of Kawai et al. and Scott and Jordan index) was studied. The results show that in the cases with fetal pathology (small for date, depression, acidosis, perinatal death) as well as in the cases with placental pathology, the values of pregnanediol were significantly decreased from the first months of pregnancy, which indicate that the placental insufficiency is a very precoz chronic process and on the other hand, that the determination of urinary pregnanediol is a good method for its detection.

Apgar Score↗
PubMed1982

Radioimmunoassay of pregnanediol concentrations in early morning urine specimens for assessment of luteal function in women.

A radioimmunoassay has been developed that employs a readily available radioligand 3H-20alpha-hydroxy-4-pregnen-3-one for the determination of pregnanediol glucuronide in urine. The unextracted steroid is assayed directly after dilution of urine with the use of an antiserum produced to a thyroglobulin conjugate of pregnanediol glucuronide. The assay has been validated, and the ability to assess luteal function by measurement of the concentration of pregnanediol glucuronide in early morning urine specimens has been demonstrated.

Adult↗
PubMed1980

[Human placental lactogen, total oestrogen and pregnanediol. a comparison of the clinical relevance on a quantitative basis (author's transl)].

of urinary total oestrogen and pregnanediol were undertaken in 65 high risk pregnancies which resulted in the delivery of 65 hypotrophic infants. The relative clinical relevance of the three examined parameters was investigated by graphical comparison of the pathological ranges with the normal ranges (degree of overlapping). In respect of hypotrophy the ratio of pregnanediol : total oestrogen : HPL is equal to 7 : 5 : 1. This shows a sevenfold improved correlation between pregnanediol and fetal weight than between HPL and fetal weight.

Estrogens↗
PubMed1977

Analysis of pregnanediol in urine: trental interference.

Patients given Trental (pentoxifyllin, Hoechst A.G.) for the treatment of vascular disorders excrete a substance which interferes with the gas chromatographic anlaysis of pregnanediol. The interfering substance appeared in urine within 4-5 h of the ingestion of 100 mg Trental, and was cleared during the next 4-10 h. The administration of 300 mg/24 h to an anovulatory woman caused an apparent increase in her output of pregnanediol of from less than 1 to 65.8 micronmol/24 h.

Chromatography, Gas↗
PubMed1988

Urinary estrone glucuronide, pregnanediol glucuronide and human chorionic gonadotrophin in threatened abortion.

Estimations of urinary estrone glucuronide, pregnanediol glucuronide and human chorionic gonadotrophin were carried out by ELISA to see their potential in predicting an abnormal outcome in cases with vaginal bleeding in early pregnancy. Reference values were set up with samples from women without bleeding in present or past pregnancies and with normal ultrasonic findings. None of the parameters were found to be sensitive enough to predict an abnormal outcome. However, predictability of an abnormal value was found to be 95% for estrone-3-glucuronide (E1G), 93% for pregnanediol glucuronide (PdG) and 87% for human chorionic gonadotrophin (hCG).

Abortion, Missed↗