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PubMed1996

Detection of adrenomedullin, a hypotensive peptide, in amniotic fluid and fetal membranes.

OBJECTIVE: Our purpose was to determine whether adrenomedullin, a multifunctional regulatory peptide involved in blood flow regulation and growth stimulation and with antimicrobial activity, was a component of amniotic fluid from second-trimester human fetus and to determine the source of this peptide. STUDY DESIGN: A prospective descriptive study was performed on 134 patients undergoing amniocentesis after genetic counseling, ultrasonography, and informed consent. Adrenomedullin expression was determined by immunocytochemical analysis, Western blot analysis, reverse transcriptase-polymerase chain reaction, and in situ reverse transcriptase-polymerase chain reaction in fetal membranes and with radioimmunoassay in amniotic fluids. RESULTS: Radioimmunoassay of the 134 amniotic fluid specimens revealed adrenomedullin-like immunoreactivity in all of them, ranging in concentration from 10 to 300 fmol/25 microliters (170 +/- 62 fmol/25 microliters). Immunocytochemical analysis, Western blot analysis, reverse transcriptase-polymerase chain reaction, and in situ reverse transcriptase-polymerase chain reaction further established the expression of adrenomedullin protein and messenger ribonucleic acid in fetal amniotic membranes, suggesting that this organ is the source of amniotic adrenomedullin. CONCLUSIONS: Our results clearly demonstrate the presence of adrenomedullin in second-trimester human amniotic fluid and adrenomedullin messenger ribonucleic acid and protein in amniotic membranes, suggesting that adrenomedullin is a hormone involved in the maintenance of normal pregnancy. Further studies with these molecular tools are in progress to determine the precise role of this hormone and whether adrenomedullin plays a role in the pathogenesis of various disorders of pregnancy.

Adrenomedullin↗
PubMed1975

Critical assessment of phospholipid measurement in amniotic fluid.

We assessed several methods of inorganic phosphate assay for their suitability in estimating phospholipids in digested extracts of amniotic fluids. The extraction and digestion procedures used for phospholipids from amniotic fluid were also examined critically. The effect of contamination by blood or obstetric cream has been examined. Accordingly, we suggest a method for measuring total phospholipids in amniotic fluids, and results of it are compared with the lecithin/sphingomyelin ratio measurement in some clinical situations.

Amniotic Fluid↗
PubMed1975

[Study of soluble proteins from human amniotic fluid by fraction chromatography (author's transl].

The nature and origin of soluble proteins from human amniotic fluid have been investigated by gel filtration chromatography on Sepharose 6B and ion-exchange chromatography on Ecteola-cellulose. Each fraction has been studied by immunoelectrophoresis. Specific antisera against serum proteins, amniotic fluid proteins, fetal proteins and salivary proteins have been used. These various antisera showed that the amniotic fluid contains maternal proteins, but also more specific proteins from fetal origin such as beta2-microglobulin, urinary mucopolysaccharides, salivary proteins and carcinoembryonic antigen. These results not only confirm that amniotic proteins are essentially from maternal origin, but prove the important fetal contribution and emphasize the importance of the fetoplacental unit in the monitoring of high risk pregnancies.

Amniotic Fluid↗
PubMed1995

Soluble tumor necrosis factor receptors and soluble interleukin-6 receptor in fetal and maternal sera, coelomic and amniotic fluids in normal and pre-eclamptic pregnancies.

The aim of this study was (a) to measure soluble tumor necrosis factor receptors (sTNF-Rs) and soluble interleukin-6 receptor (sIL-6-R) in coelomic and amniotic fluids, cord and maternal sera in pregnancy and labor, (b) to examine whether the changes in concentrations of biologically active TNF and IL-6 are related to changes in their soluble receptors, and (c) to determine if levels of soluble receptors in pre-eclamptic disorders differ from normal pregnancies at delivery. Materials collected from 206 women during pregnancy and at delivery were analyzed for soluble receptors by enzyme-linked immunosorbent assay (ELISA). All receptors were present in higher concentrations in coelomic than in the corresponding amniotic fluid. Concentrations increased in amniotic fluid from first to second trimester. The level of sIL-6-R then remained unchanged to term, but there was a decrease in the sTNF-Rs which might account for the simultaneous appearance of bioactive TNF. Labor did not affect the concentration of any receptor in amniotic fluid. In maternal serum, sTNF-Rs increased with gestational age and labor in parallel with IL-6. The origin and physiological importance of these soluble receptors are still unknown. In pre-eclamptic disorders p55 sTNF-R was elevated in maternal serum before initiation of labor compared to normal pregnancy.

Adolescent↗
PubMed1998

Changes in plasma and amniotic fluid endothelin levels during pregnancy: facts or artefacts?

OBJECTIVE: To describe the evolution of immunoreactive endothelins (irETs) in maternal peripheral plasma and in amniotic fluid at different stages of pregnancy using two different radioimmunoassays. STUDY DESIGN: Peripheral blood samples were obtained from ten non-pregnant and eighty-four pregnant patients at different stages of pregnancy, but not in labor. Amniotic fluid samples were obtained in ten patients during the second trimester of pregnancy and in twenty-two patients at term. Endothelin concentrations were assayed using two different kits: by antigenic cross-reactions, the RPA 545 assay allowed the detection of ET-1, ET-2 and big ET-1; the RPA 555 assay allowed the detection of ET-1, ET-2 and ET-3. RESULTS: Using the RPA 555 kit, no differences were observed in irET plasma levels between non-pregnant and pregnant patients whatever the gestational age. With the RPA 545 kit, the irET levels were significantly lower in pregnant patients during the first and second trimesters of pregnancy when compared to non-pregnant patients. Immunoreactive ET levels then increased significantly in the last month of pregnancy when compared to mid-pregnancy levels. In amniotic fluid, irET levels were significantly higher at term than during the early second trimester, without any difference between the two RIA kits. CONCLUSION: Our results are indicative of a differential evolution in ET isoforms during pregnancy in maternal peripheral blood. The increase in irET observed towards the end of pregnancy in maternal plasma and in amniotic fluid suggests that ET could play a role in the onset of parturition.

Amniotic Fluid↗
PubMed2002

Ultrasound estimation of amniotic fluid volume using the largest vertical pocket containing umbilical cord: measure to or through the cord?

OBJECTIVES: The purpose of this investigation was to determine the preferable method, either measuring to the umbilical cord or through the umbilical cord to the base of the pocket, of ultrasonically estimating amniotic fluid volume. SUBJECT AND METHODS: This was a prospective study carried out in singleton pregnancies undergoing a third-trimester amniocentesis. The amniotic fluid index (AFI) and single deepest pocket (SDP) were measured prior to amniocentesis. If measured spaces contained umbilical cord, measurements were made to and through the cord. Actual amniotic fluid volume was determined by the dye-dilution technique. RESULTS: One-hundred pregnancies were evaluated. Low dye-determined volume was identified in a significantly greater number of pregnancies using the AFI to the cord (7/28, 25%) compared to through the cord (2/28, 7%) (P = 0.025). The SDP technique to the cord was superior in low volumes (2/28, 7%) vs. (0/28, 0%) through the cord (although statistical significance could not be determined because there were no low through-the-cord measurements). CONCLUSIONS: For the detection of low amniotic fluid volumes, the AFI to the cord is better than through the cord. Measurement to the cord and through the cord had similar accuracy for both the AFI and SDP techniques in normal and high dye-determined amniotic fluid volumes.

Adolescent↗
PubMed1990

Cotinine concentrations in amniotic fluid and urine of smoking, passive smoking and non-smoking pregnant women at term and in the urine of their neonates on 1st day of life.

Cotinine was measured in the amniotic fluid and urine of 31 pregnant women and in the urine of their offspring. Amniotic fluid cotinine was 8 times higher in active and 2.5 times higher in passive smokers than in non-smokers. In general, amniotic fluid cotinine was considerably higher than urinary cotinine both in active and in passive smokers. Estimation of cotinine both in amniotic fluid immediately before delivery and in urine of the newborn on the 1st day of life aids in assessing the degree of prenatal exposure to tobacco smoke.

Amniotic Fluid↗
PubMed2002

ICAM-1 in maternal serum and amniotic fluid as an early marker of preeclampsia and IUGR.

OBJECTIVE: To determine if plasma and amniotic fluid levels of intercellular adhesion molecule-1 (ICAM-1) at 16 weeks' gestation could be predictive of preeclampsia or intrauterine growth retardation (IUGR). STUDY DESIGN: A retrospective analysis was undertaken in 44 serum samples stored for Down's syndrome screening at 16 weeks' gestation and 44 amniotic fluid samples obtained by midtrimester amniocentesis. RESULTS: No significant difference was found between women who subsequently developed preeclampsia or IUGR and the control group. CONCLUSION: This study failed to demonstrate that ICAM-1 may be an early serum marker of preeclampsia or an amniotic fluid marker of IUGR.

Adult↗
PubMed1979

Free and protein-bound steroids in amniotic fluid of midpregnancy.

Ninety-seven amniotic fluid (AF) specimens, 67 with male fetuses and 30 with female fetuses, were obtained by amniocentesis at 14 to 18 weeks of gestation for analysis of free and protein-bound estrone (E1), estradiol (E2), androstenedione (A), testosterone (T), and progesterone (P). Ten maternal plasma (MP) samples of comparable gestational age were analyzed for the same free and protein-bound steroids. Significantly lower total E1, E2, A, and T, but not P, levels were found in AF when compared to MP. The percentage of free steroids was higher in AF than in MP, while the percentage of specific protein-bound steroids was lower in the AF than in MP. Free (unbound) steroid levels of E1 and P were higher in AF than in MP, whereas levels of E2 were lower in AF than in MP. The free A and T levels in MP were lower than in AF with male fetuses, yet higher than in AF with female fetuses. Possible biologic activity of steroids in this fluid may depend on the dynamic balance between the free, the specific protein-bound, and the nonspecific protein-bound steroids.

Adult↗
PubMed1979

Characteristics of galactokinase and galactose-1-phosphate uridyltransferase in cultivated fibroblasts and amniotic fluid cells.

The kinetic characteristics of galactose-1-phosphate uridyltransferase and galactokinase in cultivated fibroblasts and amniotic fluid cells were investigated. The Km values of galactokinase for galactose at 2.0 mM ATP are 0.34 mM in amniotic fluid cells and 0.48 mM in fibroblasts. The Km values for ATP at 0.5 mM galactose are 1.25 mM and 2.10 mM. Transferase and galactokinase activities and protein content increase logarithmically during the growth of cultivated cells. The specific activity of both enzymes also increases and reaches a maximum level 10--15 days after subculture. The specific activity of transferase increases faster than that of galactokinase in the case of amniotic fluid cells. In the case of fibroblasts the specific activity of galactokinase increases faster than that of transferase.

Amniotic Fluid↗
PubMed1981

Comparison of amniotic fluid lung maturity profiles in paired vaginal and amniocentesis specimens.

Amniotic fluid was obtained by transvaginal amniocentesis from 27 pregnant women in the third trimester. Subsequent to amniocentesis, membranes were ruptured and vaginal pool amniotic fluid was taken by bulb aspiration. The samples were analyzed for lung maturity profile, consisting of the lecithin/sphingomyelin (L/S) ratio and the percentages of phosphatidylinositol and phosphatidylglycerol. The L/S ratio was higher in the vaginal pool than in the amniotic sac in 22% of the paired samples. Similarly, the percentage of phosphatidylinositol was greater in the vaginal pool than in the amniotic sac in 48%. Phosphatidylglycerol, however, is invariably present in the amniotic sac when it is detected in the vaginal pool. The possible usefulness of these results in the management of patients with premature rupture of membranes is discussed.

Amniocentesis↗
PubMed2003

The usefulness of ultrasound assessment of amniotic fluid in predicting adverse outcome in prolonged pregnancy: a prospective blinded observational study.

OBJECTIVE: To determine whether a single ultrasound scan at or beyond 40 weeks of gestation to detect a single deepest pool of amniotic fluid <2 cm and amniotic fluid index (AFI) <5 cm is clinically useful in the prediction of subsequent adverse pregnancy outcome. DESIGN: A prospective double blind cohort study. SETTING: A university teaching hospital delivering approximately 6000 women annually. POPULATION: One thousand and five hundred and eighty-four pregnant women at or beyond 40 weeks of gestation. METHODS: Ultrasound assessment of liquor to detect the single deepest pool of amniotic fluid and derive the AFI at or after 40 weeks of gestation. MAIN OUTCOME MEASURES: Perinatal death, meconium aspiration, birth asphyxia, intervention in labour for fetal distress, a cord arterial pH <7 and admission to the neonatal unit. RESULTS: An AFI <5 cm but not a single deepest pool <2 cm was significantly associated with birth asphyxia or meconium aspiration. An AFI <5 cm was also significantly associated with caesarean section for fetal distress in labour, a cord arterial pH <7 at delivery and low Apgar scores. Despite there being a statistically significant association with adverse outcomes the sensitivity of AFI was low at 28.6%, 12% and 11.5% for major adverse outcome, fetal distress in labour or admission to the neonatal unit, respectively. CONCLUSIONS: The AFI is superior to a measure of the single deepest pool as an assessment of the fetus at or after 40 weeks but has a poor sensitivity for adverse pregnancy outcome. Routine use is likely to lead to increased obstetric intervention without improvement in perinatal outcomes.

Adult↗
PubMed1989

A glycosylated prolactin species is covalently bound to immunoglobulin in human amniotic fluid.

Western immunoblots performed during separation of glycosylated prolactin from amniotic fluid revealed that some of the glycosylated prolactin is covalently bound to another protein. Using high performance liquid chromatography and other protein isolation techniques we have demonstrated that a glycosylated prolactin species is linked to immunoglobulin by disulfide bonds in amniotic fluid.

Amniotic Fluid↗
PubMedDate not supplied

Detection of cocaine and its metabolites in amniotic fluid and umbilical cord tissue.

The increased use of cocaine by women of child-bearing age has left many health care scientists searching for improved methods of detecting prenatal cocaine exposure. To that end, a study of the determination of cocaine and its metabolites in amniotic fluid and umbilical cord tissue was undertaken. Amniotic fluid (n = 32) and umbilical cord tissue (n = 70) specimens were collected from pregnant subjects admitted to labor and delivery at Shands Hospital at the University of Florida (Gainesville, FL). Subjects were interviewed regarding drug use during each trimester. Subjects reporting cocaine use were designated as target subjects, and those denying use were control subjects. The specimens were subjected to solid-phase extraction and analyzed for cocaine and its metabolites by gas chromatography-mass spectrometry. Cocaine analytes (predominantly benzoylecgonine) were detected in 28.1 and 18.5% of the amniotic fluid and umbilical cord tissue specimens, respectively. Other cocaine analytes frequently detected included ecgonine methyl ester and m-hydroxy-benzoylecgonine in amniotic fluid specimens and ecgonine methyl ester, norcocaine, and m-hydroxybenzoylecgonine in umbilical cord tissue specimens. This study has shown that cocaine and its metabolites are readily detected in specimens of maternal and fetal origin.

Adolescent↗
PubMed1978

Somatomedin bioactivity in serum and amniotic fluid during pregnancy.

Somatomedin bioactivity (SMA) in maternal serum, as measured by hypophysectomized rat cartilage assays, was low during early and late pregnancy but was similar to normal nonpregnant control levels near 28 weeks and at term pregnancy. At term, the mean level of SMA in cord serum was significantly less than in maternal serum, and greater than in amniotic fluid. The mean level of SMA in amniotic fluid was higher at term than in early pregnancy. No significant correlation was noted between SMA levels in amniotic fluid, maternal serum or cord serum, or between the SMA levels in these samples and the usual amniotic fluid parameters used as indices of fetal maturity.

Amniotic Fluid↗
PubMed2006

Annexin A5 levels in midtrimester amniotic fluid: association with intrauterine growth restriction.

OBJECTIVE: The purpose of this study was to determine whether amniotic fluid levels of annexin A5 (AF-AnxA5) may be associated with intrauterine growth restriction (IUGR). STUDY DESIGN: Amniotic fluid was obtained in a cohort of pregnant patients (n = 172) undergoing amniocentesis at 15 to 24 weeks' gestation and annexin A5 (AnxA5) levels were determined with enzyme-linked immunosorbent assay (ELISA). Patients who developed IUGR were compared with the remainder of the cohort. RESULTS: AF-AnxA5 levels significantly increased through gestation (P = .003). The mean level of AF-AnxA5 in IUGR patients (n = 15) was significantly higher (P = .028) than in the remainder of the cohort (n = 130). Elevated AF-AnxA5 > 20 ng/mL was associated with a relative risk of 3.5 for the development of IUGR (P = .023). CONCLUSION: AnxA5 is present in amniotic fluid and increases through gestation from 15 to 24 weeks. Elevated AF-AnxA5 levels were present in patients who developed IUGR. AF-AnxA5 may be a useful marker for identifying pregnancy abnormalities.

Adult↗
PubMed1989

Ceramide lactoside in amniotic fluid: high concentration in chorioamnionitis and in preterm labor.

The mechanisms responsible for the onset and progression of preterm labor are poorly understood. In the present study a total of 115 amniotic fluid specimens were analyzed for a lipid that has not previously been detected in amniotic fluid. This glycolipid was identified as ceramide lactoside. It was found in two-dimensional thin-layer chromatograms for evaluation of lung maturity and quantified by gas chromatography. Ceramide lactoside concentrations in amniotic fluid were low in spontaneous labor at term (1.7 +/- 0.7 nmol/ml) and in pregnancies that were not associated with spontaneous preterm labor (1.4 +/- 0.6 nmol/ml). The concentrations were high in chorioamnionitis with signs of infection (11.8 +/- 5.8 nmol/ml) and in preterm labor without clinical signs of chorioamnionitis (5.4 +/- 4.0 nmol/ml). A high ceramide lactoside (greater than or equal to 5 nmol/ml) predicted chorioamnionitis with signs of infection at a sensitivity and a specificity of 94% and 95%, respectively. A moderately high ceramide lactoside concentration (greater than or equal to 2.5 nmol/ml) predicted spontaneous preterm labor: sensitivity, 82%; specificity, 95%. Little, if any, ceramide lactoside was present in urine, vernix, normal fetal membranes, or lung effluent, whereas this glycolipid was present in large amounts in granulocytes and in inflamed fetal membranes. We propose that phagocytosing granulocytes release ceramide lactoside into amniotic fluid.

Amniocentesis↗
PubMed1969

Lesch-Nyhan mutation: prenatal detection with amniotic fluid cells.

Cells cultured from the amniotic fluid of a 22-week fetus in a heterozygote for the X-linked Lesch-Nyhan mutation, which results in neurological and developmental disorders, lacked sex chromatin and were unable to incorporate hypoxanthine. The diagnosis of a mutant male was confirmed upon birth of enzyme-deficient, hyperuricemic twin boys whose amniotic membrane cells failed to incorporate hypoxanthine.

Amniotic Fluid↗