The impact of micronized progesterone on the endometrium: a systematic review.

Postmenopausal women with an intact uterus using estrogen therapy should receive a progestogen for endometrial protection. International guidelines on menopausal hormone therapy (MHT) do not specify on progestogen type, dosage, route of application and duration of safe use. At the same time, the debate on bioidentical hormones including micronized progesterone increases. Based on a systematic literature review on micronized progesterone for endometrial protection, an international expert panel's recommendations on MHT containing micronized progesterone are as follows: (1) oral micronized progesterone provides endometrial protection if applied sequentially for 12-14 days/month at 200 mg/day for up to 5 years; (2) vaginal micronized progesterone may provide endometrial protection if applied sequentially for at least 10 days/month at 4% (45 mg/day) or every other day at 100 mg/day for up to 3-5 years (off-label use); (3) transdermal micronized progesterone does not provide endometrial protection.

Basics of sterile compounding: compounding by weight versus compounding by volume.

In compounding small volumes of sterile preparations, it is generally acceptable to use volumetric measurements. However, when compounding larger volumes or when the sterile preparation has a low concentration, it is best to compound by weight for greater accuracy. This article provides the procedures that can be followed when compounding by weight as compared to volume and provides an example of the steps involved in compounding 18.5 liters of progesterone 100 mg/mL injection.

The standardization of nonsterile compounding: a study in quality control and assessment for hormone compounding.

Sterile and nonsterile compounding of medication has attracted much attention over the last few years due to the onset of various infections and negative compounding practices. This paper reports on the standardization of compounded hormones utilizing the Wiley Protocol, which provides nonsynthetic bioidentical estradiol, progesterone, dehydroepiandrosterone, and testosterone in a transdermal topical cream base for women and men in a standardized dosing regimen. Here, we present data from 2008 through 2012, which details the process of standardization and quality testing of the hormones through submission of random compounded samples for quality control and assessment. Pharmacies delivering the Wiley Protocol were required to follow the same compounding formulation, as well as submit random samples for quarterly testing. Sample concentrations were tested using high-performance liquid chromatography. We found that pharmacies that submitted samples had a 91% passing rating with a percent of target of 98.6% +/- 8.4%. It was also determined that pharmacies that prepared more compounded cream had a higher passing rating than those that prepared limited quantities. We found that standardization across multiple pharmacies could be achieved through quarterly testing of submitted samples by a third-party laboratory when following necessary procedures as defined by the Wiley Protocol. It was also determined that experience and training were a critical factor in the mixing of compounded prescriptions, with high consistency and accuracy providing patient safety.

Progesterone level and progesterone/estradiol ratio on the day of hCG administration: detrimental cutoff levels and new treatment strategy.

OBJECTIVE: To identify if there are certain cutoff levels for P and or the P/E(2) ratio on the day of hCG that would be defined as detrimental for occurrence of pregnancy in women with normal ovarian reserve undergoing cleavage-stage embryo transfer (ET). Secondarily, to determine if these same cutoffs might have the same potential negative effect in women undergoing blastocyst ET. DESIGN: Prospective cohort study including two randomized cohorts. SETTING: Private and university fertility centers. PARTICIPANT(S): A total of 240 women undergoing long agonist protocol with at least four grade 1 day 3 embryos. INTERVENTION(S): Women were randomized in a 1:1 ratio to undergo day 3 or day 5 embryo transfer. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate (CPR) was the primary outcome. RESULT(S): Using receiver operator characteristics, cutoffs for P and P/E(2) ratio were 1.5 ng/mL and 0.55, respectively. Patients with P ≤ 1.5 ng/mL and P/E(2) ≤ 0.55 undergoing cleavage-stage ET had higher CPR. Using multiple regression, P/E(2) ratio was the only independent predictor for pregnancy. The P and P/E(2) cutoffs were not correlated with CPR in blastocyst transfers. CONCLUSION(S): Progesterone >1.5 ng/mL and P/E(2) >0.55 affect the CPR in women undergoing cleavage-stage, but not blastocyst ET. P/E(2) ratio is the only independent prognosticator for cycle outcome in women undergoing cleavage-stage ET.

Development of dual-enzyme-based simultaneous immunoassay for measurement of progesterone and human chorionic gonadotropin.

The development of a simultaneous multianalyte immunoassay for the detection of progesterone and human chorionic gonadotropin (hCG) in serum is described. In this simultaneous multianalyte assay, two different enzymes, viz. horse radish peroxidase (HRP) and alkaline phosphatase (ALP), were used as markers. To the simultaneous immobilized progesterone and hCG antibody microwells, 50 microL of different concentrations of combined standards or serum samples was added in duplicate and then 100 microL of combined conjugate reagent, composed of 17-alpha-OH-P-ALP and hCG-biotin was added to all the wells and incubated for 1h at 37 degrees C. After incubation, the contents of the wells were decanted and washed thoroughly with running tap water. After washing, 100 microL alkaline phosphatase substrate along with streptavidin-horseradish peroxidase was added to all the wells and incubated for 0.5 h at 37 degrees C. After incubation, the developed color was measured at 405 nm. The absorbency at this stage provides the result for the progesterone assay. The contents of the wells were decanted and washed. In the next step, 100 microL of tetramethylbenzidene/H2O2 reagent was added to all the wells. After 15 min of incubation, 100 microL of 0.5 M H2SO4 was added to all the wells and the color was read at 450 nm. The absorbency at this stage provides the result for the hCG assay. Sensitivity of the progesterone and hCG assays were 0.118 ng/ml and 0.124 IU/ml respectively. Intra- and inter assay percentage coefficients of variation ranged from 1.8 to 7.1 and 9.1 to 11.5 for progesterone and from 2.1 to 10.4 and 7.2 to 11.3 for hCG. There was good correlation between the discrete and the simultaneous assays. For progesterone assay, R2 was 0.99 and for hCG R2 was also 0.99. The developed dual assay for progesterone and hCG may be useful for the diagnosis of abnormal pregnancies such as miscarriages and ectopic pregnancies.

Studies on the influence of the structural modifications in the tracer on the immunoassay of progesterone.

The main objective of the present study was to examine the influence of different bridges in radioiodinated tracers on the assay performance of progesterone using antibodies. Three homologous and two heterologous immunoassay systems for the measurement of progesterone in human serum are described. Using an antiserum raised against progesterone-11alpha-hemisuccinate-bovine serum albumin (BSA), assays with homologous radioligands, namely progesterone-11alpha-hemisuccinate-125I-tyrosine methyl ester (TME) and progesterone-11alpha-hemisuccinate-125I-histamine, heterologous bridge radioligand, namely progesterone-11alpha-hemiphthalate-125I-TME, and a heterologous site radioligand namely progesterone-3-(O-carboxymethyl) oxime (CMO)-125I-histamine were optimized. A homologous assay system, using antiserum raised against progesterone-3-carboxymethyl oxime-BSA and progesterone-3-CMO-125I-histamine as the radioligand was also optimized to develop a radio-immunoassay (RIA) for serum progesterone. Amongst the two homologous radioligands, viz., progesterone-11alpha-hemisuccinate-125I-histamine and the corresponding TME conjugate tracer, the former yielded a standard curve with a higher slope (-0.6) as compared to the latter (-0.5). The heterologous bridge system with progesterone-11alpha-hemiphthalate-125I-TME resulted in a more sensitive assay (slope of -0.8) than the homologous tracers, whilst the heterologous site radioligand, viz., progesterone-3-CMO-125I-histamine gave the most sensitive assay (slope of -1.2). The homologous assay with antiserum against progesterone-3-CMO-BSA and progesterone-3-CMO-125I-histamine tracer gave a standard curve having a slope of -0.97. The two antibodies developed against progesterone, viz., progesterone-11alpha-hemisuccinate-BSA and progesterone-3-CMO-BSA were characterized for their titre, sensitivity, and specificity. Considering the slope, sensitivity, cross-reactivity, and the quality of tracer, the assay system using antiserum against progesterone-11alpha-hemisuccinate-BSA and progesterone-3-CMO-125I-histamine was found to be suitable for the development of RIA for serum progesterone. The bridges used in an immunogen for production of antibodies, as well as in the preparation of tracer, have a great influence on the assay characteristics.

A comparison of the effects of progesterone sponges and ear implants, PGF2alpha, and their combination on efficacy of estrus synchronization and fertility of Mashona goat does.

Efficacy of estrus synchronization and fertility after synchronization of 60 multiparous Mashona goat does using intravaginal progesterone (P4) sponges (Group 1), norgestomet ear implants (Group 2), cloprostenol (Group 3), or a combination of P4 sponges and cloprostenol (Group 4) was compared with untreated does (Group 5). At the end of treatments, all does were mated to intact fertile bucks for 21 d. The number of does bred within 11 to 96 h was significantly higher (P < 0.05) in the treated groups than the untreated control, with rates of 80, 80, 64, 67 and 30% for Groups 1 to 5, respectively. There were no differences (P > 0.05) among treated does. Kidding rates ranged from 64 to 83% but were not different (P > 0.05) between groups. Prolificacy and overall fecundity were similar (P > 0.05) among the groups. The results indicate that all 4 treatment methods were effective in synchronizing estrus and that none of the methods affected overall fertility of the does.

[The Progesterone Reference Standard (Control 931) of the National Institute of Health Sciences].

Raw progesterone material was tested for preparation of the "Progesterone Reference Standard (Control 931)". Analytical data obtained were as follows: melting point, 131.6 degrees C; infrared spectrum, the same as that of the JP Progesterone Reference Standard; optical rotation, [alpha]D20 = +181.1 degrees; thin-layer chromatography, no impurity was detected; high-performance liquid chromatography (HPLC), trace amounts of two impurities were detected; loss on drying, 0.23%; assay results, 99.4% by UV spectrophotometry in the JP XII and 100.5% by HPLC. Based on the above findings, the raw material was authorized as the JP Progesterone Reference Standard (Control 931).

Development, validation, and certification by isotope dilution gas chromatography-mass spectrometry of lyophilized human serum reference materials for cortisol (CRM 192 and 193) and progesterone (CRM 347 and 348).

The Community Bureau of Reference of the European Communities has produced four batches of lyophilized serum Certified Reference Materials, two for cortisol (CRM 192 and 193) and two for progesterone (CRM 347 and 348). For cortisol, one of the pools consisted of serum from healthy blood donors, whereas the second batch was supplemented with pure cortisol. The progesterone Reference Materials contained only endogenous hormone concentrations. Assessment of vial-to-vial variability in the cortisol and progesterone concentrations showed no between-sample inhomogeneity, and the materials were stable. The quality of the materials was therefore considered sufficient for certification of the values for the cortisol and progesterone concentrations by a collaborative study involving several laboratories from the European Communities, using isotope dilution gas chromatography-mass spectrometry. Inaccuracy in reconstitution of the lyophilized materials was less than 0.3%; imprecision of sampling was less than 0.2%. For determinations of cortisol and progesterone concentrations, the mean within-laboratory coefficients of variation (CVs) were 1.76% (CRM 192), 1.19% (CRM 193), 1.64% (CRM 347), and 1.75% (CRM 348). The between-laboratory CVs were greater: CRM 192, 1.79%; CRM 193, 1.48%; CRM 347, 2.08%; and CRM 348, 2.16%. The concentrations in the reconstituted Reference Materials were certified to be 273 nmol/L in CRM 192 and 763 nmol/L in CRM 193 for cortisol and 10.13 nmol/L in CRM 347 and 40.3 nmol/L in CRM 348 for progesterone. Uncertainties at the 0.95 confidence level--6 (CRM 192), 14 (CRM 193), 0.21 (CRM 347), and 1.0 nmol/L (CRM 348)--were considered compatible with the intended use of the materials.

[Serum concentrations of delta 5C21 steroids during pregnancy and at delivery].

In order to study the changes of C21-steroid levels which included Pregnenolone (P5), 20 alpha dihydropregnenolone (20P5), 16 alpha hydroxypregnenolone (16P5), progesterone (P4) and 20 alpha dihydroprogesterone (20P4) in maternal peripheral blood during pregnancy and at delivery, these steroids were measured by GC-MASS with application of deuterated steroids as internal standard. The accuracy of GC-MASS method of these steroids was satisfactory with C.V. value of less than 6%. Total delta 5C21 steroid concentrations in course of pregnancy and at delivery were as follows; P5 (mean +/- S.D. ng/ml): 66.6 +/- 36.2 (1st trimester), 80.9 +/- 24.6 (2nd trimester), 147.7 +/- 30.1 (3rd trimester) and 299.7 +/- 178.3 ng/ml (at delivery), 20Ps: 212.6 +/- 102.5, 143.4 +/- 53.9, 248.9 +/- 58.8, 563.4 +/- 198.2 ng/ml, 16P5: 8.6 +/- 8.6, 8.1 +/- 5.2, 124.3 +/- 40.3, 378.5 +/- 180.0 ng/ml, respectively. P4 (43.0 +/- 28.0 ng/ml) and 20P4 (8.0 +/- 4.0 ng/ml) in 1st trimester showed gradual increase to maximum level (P4: 138.2 +/- 30.1 ng/ml, 20P4: 105.4 +/- 21.6 ng/ml) at pre-pain period, afterward decreased rapidly (P4: 70.9 +/- 23.2 ng/ml, 20P4: 59.8 +/- 19.3 ng/ml) at delivery. P5, 20P5 and 16P5 levels were found to be significantly higher in umbilical artery (UA) as well as in umbilical vein (UV) than those in maternal vein (MV) regardless of labor pain. P4 and 20P4 did not show any differences in MV regardless of labor pain. P4 in UV (pain+) and 20P4 in UA (pain-), however, showed significantly higher than P4 in UV (pain-) and 20P4 in UA (pain+). P5, 20P5, 16P5 and 20P4 levels were significantly lower in the case of anencephalic pregnancy (ANC) at 3rd trimester than in normal pregnancy, especially 16P5 levels (22.2 +/- 5.0 ng/ml) showed 1/5 of those in normal pregnancy. From the results obtained above, it is suggested that these delta 5C21 steroids are actively produced in the feto-placental unit in the course of pregnancy. The levels of these steroids reached maximum at delivery, but the levels of P4, 20P4 decreased toward delivery after maximum levels were shown in the stage of pre-labor pain. No significant difference of P4 level in the case of ANC suggested that P4 production correlated with placenta as well as maternal and fetal precursor. Decreasing of 20P4 and P4 level after the stage of pre-labor pain suggested that activity of 3 beta hydroxysteroid dehydrogenase was reflected by uterine contraction during labor.