High concentrations of CA 125 in uterine flushings: influence of cause of infertility and menstrual cycle day.

Uterine flushings were obtained under transvaginal ultrasonographic control from 132 women presenting for investigation and treatment of infertility. Levels of CA 125 were measured by radioimmunoassay and results expressed in relation to the total protein concentration of the same flushings. CA 125 was detected in uterine fluid at levels higher than those previously reported in peripheral blood. Uterine fluid CA 125 concentrations varied throughout the menstrual cycle, being highest in the mid-follicular phase (days 6 to 10). Uterine fluid CA 125 concentrations may reflect endometrial secretion of this protein more directly than serum levels. CA 125 concentrations did not vary according to the cause of infertility but further work in larger numbers of women is required.

Delayed luteo-placental shift of progesterone production in IVF pregnancy.

OBJECTIVE: To observe absolute and relative levels of progesterone, 17 alpha-hydroxyprogesterone (17-OHP) and human chorionic gonadotrophin (hCG) in in vitro fertilization (IVF) pregnancies after withdrawal of luteal support. METHOD: Single blood samples were obtained from 41 pregnant women following IVF treatment and 43 normal pregnant women at various weeks gestation within the first trimester. Progesterone, 17-OHP and hCG were measured by immunoassay. RESULTS: Serum levels of progesterone, but not of hCG, in IVF pregnancies were significantly greater than in normal pregnancies up to 8 weeks post-conception, despite discontinuing luteal support 2 weeks after conception. The ratio of progesterone to 17-OHP, a predominantly ovarian product, in normal pregnancies rose between 4 and 9 weeks but did not change over the same period in IVF pregnancies. CONCLUSION: The luteal contribution to maternal serum levels of progesterone is much higher in IVF pregnancies compared with normal pregnancies. This is sustained throughout the first trimester without the need for luteal support and obscures the placental contribution of progesterone for much longer than in normal pregnancies. Progesterone or hCG supplements may therefore be unnecessary in IVF pregnancy.

Cross-reaction with luteinizing hormone beta-core is responsible for the age-dependent increase of immunoreactive beta-core fragment of human chorionic gonadotropin in women with nonmalignant conditions.

BACKGROUND: The beta-core fragment of human chorionic gonadotropin (hCGbetacf), also termed "beta-core" and urinary gonadotropin peptide (UGP), has been reported to be present in the urine of healthy women and to increase in concentration after menopause. This could reflect cross-reaction with the equivalent metabolite of luteinizing hormone (LH), the beta-LH-core. METHODS: We measured immunoreactive LH, hCG, free alpha-subunit, and free beta-subunit hCG (hCGbeta), as well as beta-core, using the S504 RIA and Triton UGP enzyme immunoassay in 274 urine samples from women with nonmalignant gynecological conditions. The molar cross-reaction of each assay with purified beta-LH-core was determined. RESULTS: Cross-reaction with beta-LH-core was 100% in the LH and the S504 beta-core assay, 5% in the Triton UGP assay, and <0.1% in the hCG, free alpha-subunit, and free hCGbeta assays. Median urine concentrations of all analytes showed an age-dependent increase. LH and free alpha-subunit concentrations were approximately 10(3) pmol/mol creatinine; hCG and S504 beta-core were approximately 10(2) pmol/mol creatinine; free hCGbeta and Triton UGP beta-core were in the tens of pmol/mol creatinine. The S504 beta-core concentrations were 10% of those of LH. S504 beta-core was strongly correlated with LH, but not with hCG or with free hCGbeta (LH, r2 = 0.45; hCG, r2 = 0.26; free hCGbeta, r2 = 0.03). The concentrations of beta-core detected by the Triton UGP assay, which has a 5% cross-reaction with beta-LH-core, were 2% of LH and 5% of the S504 beta-core concentrations. Triton UGP values correlated strongly with LH concentrations, but less well with S504 beta-core, intact hCG, and free hCGbeta (LH, r2 = 0.44; S504 beta-core, r2 = 0.33; hCG, r2 = 0.32; free hCGbeta, r2 = 0.19). CONCLUSIONS: Immunoreactive beta-core in women free of malignancies reflects cross-reaction with concentrations of the metabolite of LH, beta-LH-core, within the health-related reference interval.

Measures of urine concentration in maternal urine screening for Down syndrome.

A study was carried out to assess eight methods of normalizing the level of urinary beta-core human chorionic gonadotropin (hCG) for variable urine concentration. We compared the standard approach--creatinine determination by the Jaffe method--with high performance liquid chromatography (HPLC) measurement of creatinine, osmolarity and optical density at five wavelengths. Urine samples were included from a total of 472 women with unaffected singleton pregnancies at 15 weeks' gestation. The median beta-core hCG value was determined for each decile group when the results were ranked in turn according to the different measures of urine concentration. Creatinine using the Jaffe method had a much stronger relationship with median beta-core hCG than the other measures. Linear regression across the decile groups gave an R2 value for Jaffe of 0.85 compared with HPLC of 0.53, osmolarity of 0.52, optical density at 405 nm of 0.72, at 450 nm of 0.57, at 490 nm of 0.33, at 570 nm of 0.34 and at 630 nm of 0.33. We conclude that when screening with urinary beta-core hCG measuring creatinine appears to be an adequate method of allowing for variable urine concentration.

Concentrations of placental protein 14 in uterine flushings from infertile women: validation of the collection technique and method of expression of results.

Concentrations of various proteins in uterine flushings have been described as a direct method for assessment of the secretory activity of the endometrium. We investigated levels of the endometrial protein known as placental protein 14 (PP14) in flushings obtained from 271 infertile women. Under transvaginal ultrasonographic control, 2 ml of 0.154 M sodium chloride solution were injected into the uterine cavity and re-aspirated, five times. In contrast to previous studies the recovered volume of each flushing was not consistent (range: 0.05-2.1 ml); the volume varied significantly between serial samples obtained from an individual (P = 0.02, one-way ANOVA), different cycle days (P < 0.0001, one-way ANOVA) and women with bilaterally blocked versus patent Fallopian tubes (P < 0.05, Student's t-test). Concentrations of PP14 showed a better correlation with protein content (r = 0.506, P < 0.0001) than with the recovered volume (r = 0.087, P = 0.095). We therefore corrected PP14 concentrations for total protein content as an indicator of the efficiency of the flushing process. Corrected PP14 concentrations varied significantly relative to time since the onset of menstruation (P = 0.001, Kruskal Wallis ANOVA) with higher levels on days 1-8, as previously observed in plasma samples. No significant difference in PP14 levels was found with different causes of infertility. This study shows that uterine flushing is not a consistent process in women with differing physical characteristics and at varying times throughout the menstrual cycle.

IGFBP-1 in the placenta, membranes and fetal circulation: levels at term and preterm delivery.

Samples of maternal blood, amniotic fluid, placenta, fetal membranes and umbilical venous blood were collected from 59 women at vaginal delivery (32-41 weeks gestation) and 15 women at delivery by Caesarean section (37-41 weeks gestation). Umbilical vein levels of IGFBP-1 were significantly lower in deliveries prior to the onset of labour (elective Caesarean section) than those during normal vaginal delivery. These levels were, in turn, significantly lower than those delivered by emergency Caesarean section. This difference was not seen in any of the other tissues examined. Concentrations of IGFBP-1 were lower in placenta and membrane extracts from preterm deliveries than in term deliveries. This difference was not observed in maternal or fetal serum or in amniotic fluid. This study confirms that the fetal membranes are a major source of IGFBP-1 and that fetal circulating levels are raised where there is evidence of fetal hypoxia. The absence of a comparable rise in levels in placenta, membranes or amniotic fluid suggests that the origin of this increase is from fetal tissue.

Lack of evidence for the production of interferon-alpha-like species by the cultured human pre-embryo.

The aim of this study was to determine if the human pre-embryo produces a substance similar to the trophoblast interferon secreted by ruminant trophoblasts. Human embryos surplus to in-vitro fertilization (IVF) treatment were cultured up to 14 days following IVF. Viable cultures were determined by microscopic examination and by assay of the culture medium for human chorionic gonadotrophin (HCG). Four stages of development were visually identified: pre-blastocyst, unhatched, part-hatched and fully hatched blastocyst. HCG was detected in medium which had contained the more developmentally advanced embryos. A total of 62 samples were assayed for human interferon-alpha (IFN-alpha), including all cultures presumed viable. None contained detectable IFN-alpha immunoreactivity. Out of 14 candidate samples subjected to cytopathic effect reduction assay, none contained antiviral activity. We suggest that a trophoblast-derived interferon, unlike HCG, does not play a significant role in the maternal recognition of pregnancy in humans.

Potential sources of artefact in the co-axial bioassay.

The apparent release of relaxant activity from airway epithelium (epithelium-derived relaxing factor, EpDRF) has been examined in a co-axial bioassay system. The endothelium-denuded rat aorta, placed inside either the epithelium-intact guinea-pig trachea or rabbit bronchus relaxed in response to acetylcholine. In a modification of the standard preparation, the airway was slit longitudinally and immobilised inside a silicone rubber tube. Under these conditions, the acetylcholine-induced relaxation was abolished. Under the conditions of the co-axial bioassay, the oxygen tension in the lumen of either airway tube was lower than that of the bathing fluid. Upon addition of acetylcholine at concentrations which caused relaxation in the co-axial bioassay, the oxygen tension inside the epithelium-intact, but not the epithelium-denuded guinea-pig trachea was depressed to levels which would have affected the contractile response of a rat aorta. We suggest that the assay of relaxant activity from airways using co-axial preparations may be complicated by changes in volume and oxygen tension in the lumen of the donor airway and discuss how such problems might be avoided.

Central inhibition by gamma-aminobutyric acid and muscimol of the release of vasopressin and oxytocin by an osmotic stimulus in the rat.

1. In water-loaded rats under ethanol anaesthesia, the injection of 2-4 microliters 1.54M NaCl solution (hypertonic saline:HS) into a lateral cerebral ventricle (i.c.v.) produced an antidiuretic and a pressor response, together with increased urinary excretion of vasopressin and 'oxytocin-like radioimmunoreactivity' (OLRI). In lactating rats HS also produced a milk-ejection response which was shown to be due to the release of oxytocin. 2. The injection of 20-40 micrograms gamma-aminobutyric acid (GABA) or 40-80 ng muscimol i.c.v. 2 min before HS inhibited the antidiuretic, pressor and milk-ejection responses and reduced the urinary excretion of vasopressin and OLRI. 3. The pressor response to HS was abolished by a ganglion blocking agent but it was not reduced by a vasopressin antagonist. After the antagonist, the antidiuretic response to HS was abolished and the pressor response was accompanied by a diuresis both of which were blocked by muscimol. 4. The threshold dose of HS for an antidiuretic response was 4-8 times higher on injection into the cisterna magna (i.cist.) than when injected i.c.v. GABA, i.v. or i.cist, did not inhibit the response to HS i.c.v. 5. The results confirm other evidence that, in the rat, in contrast some other species, an osmotic stimulus causes release of both vasopressin and oxytocin. This release is blocked by GABA and muscimol. These drugs and HS act at a site reached not from the subarachnoid space but from the cerebral ventricles, probably the hypothalamus. The pressor response to HS under the experimental conditions used is due entirely to central sympathetic stimulation and this effect, as well as the release of vasopressin and oxytocin, is blocked by muscimol.