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Capillary gas chromatographic-mass spectrometric profiles of trimethylsilyl derivatives of organic acids from amniotic fluids of different gestational age.

Amniotic fluid from different gestational age patients was partitioned into neutral, acidic and basic fractions. The organic acids were trimethylsilylated and analyzed by glass capillary gas chromatography-mass spectrometry. A marked difference in the level of hippuric acid was observed between samples from early (15-22 weeks) and late (30-38.5 weeks) pregnancy. This difference probably reflects the degree of maturity in the fetal liver and kidney. The procedures establish amniotic fluid profiles of substances of varying gestational age and should be useful in determining alterations caused by diseases.

Amniotic Fluid↗

[Determination of the acetylcholinesterase activities of the amniotic fluid on a centrifugal analyzer].

Amniotic fluid cholinesterase (ChE) activity measurements are realized to detect pre-natal neural tube defects. Their determinations on centrifugal analyzer are described. Total ChE activity is assayed at 412 nm with dithiobis-(nitrobenzoic-acid). Acetylcholinesterase activity is measured in the presence of ChE inhibitor, 14 X 10(-6)M lysivane; our results about the inhibitor concentration confirm the previous studies of Dale (Clin. Chim. Acta, 1977, 77, 407-413, Lancet, 1980, ii, 975) in opposition to those of Brock (Lancet, 1981, i, 95). Non NTD amniotic fluids from 92 women between the 17th and 20nd weeks of pregnancy were used to establish normal values. The technique proposed is rapid, accurate and needs only 40 microliters sample.

Acetylcholinesterase↗

Fetal lung maturity evaluation with fluorescence polarization of the amniotic fluid.

Fluorescence polarization of the amniotic fluid from 39 high risk pregnancies requiring preterm delivery was measured in order to assess the maturity of the fetal lung. The study population included 15 cases of intrauterine growth retardation, ten maternal hypertension, five maternal Hodgkin's disease, three placenta previa, two fetal malformation, two polyamnios, one untreated diabetes, one maternal nephropathy. All patients underwent a single amniocentesis before deciding whether to deliver a preterm baby and FP of the amniotic fluid was done within two hours from amniocentesis. In five cases this was > 0.311, the cut-off limit taken as an indicator of fetal pulmonary status, and three of these developed respiratory distress syndrome. In 34 cases FP values were < or = 0.311; in spite of the apparent lung maturity two of these newborns developed respiratory distress syndrome. On the basis of these results the FP sensitivity was calculated as 60%, specificity 94% and the overall accuracy 90%.

Amniotic Fluid↗

Epithelial cell-derived neutrophil-activating peptide-78 is present in fetal membranes and amniotic fluid at increased concentrations with intra-amniotic infection and preterm delivery.

Intra-amniotic secretion and abundance of epithelial cell-derived neutrophil-activating peptide (ENA)-78, a potent chemoattractant and activator of neutrophils, was studied in the context of term and preterm parturition. Staining of ENA-78 immunoperoxidase was localized predominantly to chorionic trophoblasts and amniotic epithelium in term and preterm gestational membranes, with weaker and less consistent staining in decidual cells. The abundance of ENA-78 in membrane tissue homogenates was significantly increased ( approximately 4-fold) with term labor in amnion (n = 15), and with preterm labor ( approximately 30-fold) in amnion and choriodecidua (n = 31). In amnion tissue homogenate extracts, ENA-78 levels were positively correlated with the degree of leukocyte infiltration (r2 = 0.481). In amniotic fluids, median ENA-78 levels from pregnancies with preterm labor without intra-amniotic infection were significantly lower (P < 0.01 by ANOVA) than those from pregnancies with preterm deliveries with infection; levels in samples derived from term pregnancies were similar before and after labor. Production of ENA-78 by amnion monolayers was stimulated in a concentration-dependent fashion by both interleukin-1beta and tumor necrosis factor alpha. Production of ENA-78 by choriodecidual explants was increased modestly after 2-4 h of exposure to lipopolysaccharide (5 microg/ml). An immunoreactive doublet ( approximately 8 kDa) was detected in choriodecidual explant-conditioned media by immunoblotting. We conclude that ENA-78, derived from the gestational membranes, is present in increased abundance in the amniotic cavity in response to intrauterine infection and, hence, may play a role in the mechanism of infection-driven preterm birth and rupture of membranes secondary to leukocyte recruitment and activation.

Amniotic Fluid↗

Amniotic fluid B12 and folate levels associated with neural tube defects.

Amniotic fluid levels of B12 and folate in neural tube defect (NTD) affected pregnancies were compared with the weekly group mean +/-SD changes in amniotic fluid B12 and folate levels of 10 unaffected pregnancies each week between 15 and 20 weeks' gestation age. Comparison was by analysis of variance (ANOVA) and Pearson's correlation to B12 and folate levels and to the NTD samples of corresponding gestation age. Amniotic fluid B12 and folate decreased 67 and 62%, respectively, between 15 and 20 weeks' gestation in the unaffected pregnancies, associated with an increase in amniotic fluid volume. The mean +/-SD B12 and folate of the NTD affected pregnancies (308+/-156 pg/mL and 3.1+/-1.6 ng/mL) were below the mean +/-SD B12 and folate of the total population of unaffected pregnancies (453+/-155 pg/mL and 3.9+/-1.2 ng/mL). The correlation between gestation age and amniotic fluid B12 was -0.9914 (p< or =.0001) and -0.9599 (p< or =.002) for amniotic fluid folate. The B12 levels of the affected pregnancies were below the range of unaffected pregnancies in four of the nine affected pregnancies, and folate levels in two of the nine affected pregnancies.

Amniotic Fluid↗

Dye-determined amniotic fluid volume and intrapartum/neonatal outcome.

OBJECTIVE: To ascertain if a dye-determined amniotic fluid volume was predictive of intrapartum and perinatal outcome. MATERIALS AND METHODS: The low and normal amniotic fluid volumes (< 5th percentile and > or =5th percentile for gestational age) and the raw dye-determined amniotic fluid distributions were correlated with 10 clinical outcome measures in 74 pregnancies. RESULTS: In this observational study, median gestational age at delivery was 36 weeks (range 26 to 41) and 16 deliveries were for fetal distress (14 Cesarean and two forceps). There were no differences between the outcomes of pregnancies with low and normal amniotic fluid volumes for any of the clinical outcomes (variable decelerations influencing delivery, p=0.381; late decelerations, p=0.875; Cesarean births for fetal intolerance of labor, p=0.259; intrauterine growth restriction, p=0.998; or umbilical cord arterial pH< 7.2, p=0.259). Analogous results were obtained when the gestational age-adjusted amniotic fluid volumes were compared directly between the pregnancies with normal and abnormal outcomes. There was no difference between the mean amniotic fluid volumes in those pregnancies with variable decelerations influencing delivery (p=0.287), late decelerations (p=0.555), Cesarean births for fetal intolerance of labor (p=0.310), intrauterine growth restriction (p=0.267) or umbilical cord arterial pH< 7.2, and the pregnancies without these intrapartum events. Reduced variability was more commonly observed in pregnancies with higher amniotic fluid volumes (p=0.038, 771 ml, 95% CI 468 to 1269, compared to those without normal variability 444 ml, 95% CI 374 to 526). CONCLUSIONS: Dye-determined amniotic fluid volume does not appear to be predictive of adverse intrapartum and neonatal outcome.

Adolescent↗

Neural tube defects and elevated homocysteine levels in amniotic fluid.

OBJECTIVE: Our purpose was to study maternal blood and amniotic fluid concentrations of homocysteine and relevant vitamins in relation to neural tube defects. STUDY DESIGN: Concentrations of total homocysteine, folate, and vitamins B12 and B6 were measured in maternal blood and amniotic fluid of 27 women carrying a fetus with a neural tube defect and 31 control women carrying a healthy fetus. RESULTS: The mean total homocysteine concentration in amniotic fluid of the study group was significantly higher than that of the control group. The mean concentrations of total homocysteine in blood and the vitamins folate, B12, and B6 in, respectively, blood and amniotic fluid were not significantly different between the groups. The mean concentrations of homocysteine and vitamin B6 were significantly lower in amniotic fluid than in blood in both groups, whereas vitamin B12 in amniotic fluid was higher than in blood. CONCLUSION: These results support the hypothesis that at least the cause of a subset of neural tube defects could reside in a primary or secondary maternal or fetal derangement of homocysteine metabolism.

Adult↗

The effect of meconium staining of amniotic fluid on the growth of Escherichia coli and group B streptococcus.

OBJECTIVE: To compare the effect of meconium staining on the growth rate of Escherichia coli and group B streptococci (GBS) in amniotic fluid. STUDY DESIGN: Sterile meconium was added in different concentrations to pooled sterile amniotic fluid obtained from term mothers. Meconium concentration was equal to: 1, 1.5, 3, 6, and 12 mg meconium/ml amniotic fluid. Amniotic fluid was quantitatively inoculated with E. coli and GBS type II and type III organisms (10(3) organisms/ml). Clear amniotic fluid and Todd-Hewitt broth served as controls. Growth rates of organisms were measured at 2, 4, 6, 8, and 24 hours after inoculation. RESULTS: Clear amniotic fluid was completely inhibitory for growth of E. coli even after 24 hours of incubation. In contrast, after 6 hours of incubation, significantly increased growth of GBS occurred (10(5) organisms/ml) (p<0.0001). The inhibitory effect on E. coli was observed even with meconium concentration of up to 1.5 mg/ml. In contrast, even the smallest concentration of meconium (1 mg/ml) resulted in a 2-log fold increase of GBS within 4 hours. The more rapid growth rate of GBS compared to E. coli persisted even at moderate staining of amniotic fluid (6 mg/ml) in the first 8 hours of incubation (p<0.005). In the presence of thick meconium (12 mg/ml), during the first 6 hours of incubation, the growth rates of GBS and E. coli were nearly similar. CONCLUSIONS: GBS (type II and III) growth, in contrast to E. coli, was less inhibited by amniotic fluid, occurred at a more rapid rate, and was enhanced at lower concentrations of meconium. As such, the presence of even light meconium staining in cases of rupture of membranes of even less than 6 hours in a mother who is a GBS carrier should be considered as a risk factor for the development of perinatal GBS infection.

Amniotic Fluid↗

[Clinical application of ultrasonographic indexes for assessment of amniotic fluid volume].

OBJECTIVE: To find the favorable interval of assessment of amniotic fluid volume and the optimal standard for oligohydramnios. METHODS: To determine actual amniotic fluid volume (AAFV) by collecting amniotic fluid during cesarean section and compare it with maximum vertical pocket (MVP) and amniotic fluid index (AFI) in different interval between cesarean section and measurement and test the sensitivity and specificity of MVP and AFI of different standard. RESULTS: The ultrasonographic results were divided into three groups according to the interval between measurement and delivery: within 24 hours group, 24-96 hours group and longer than 96 over hours group. The results of the within 24 hours and 24-96 hours groups were positively correlated with AAFV, while the results of the group longer than 96 hours were not statistically significant. The correlation coefficients were 0.840 and 0.869 for MVP, 0.789 and 0.874 for AFI respectively. The receiver-operator characteristic curve showed that MVP < 4 cm and AFI < 10 cm were superior to any other standards to scan oligohydramnios. The sensitivity of these standards was 0.76 and 0.61, and the specificity was 0.83 and 0.91. CONCLUSION: It is highly recommended to estimate the amniotic fluid of high risk pregnant women, women with MVP < 4 cm or AFI < 10 cm and with a pregnancy week > 40 weeks should be monitered by ultrasonography every four days.

Amniotic Fluid↗

The impact of amniotic fluid volume assessed intrapartum on perinatal outcome.

OBJECTIVE: Our purpose was to determine the value of routine intrapartum amniotic fluid volume assessment on perinatal outcome. STUDY DESIGN: Patients admitted for labor and delivery who were ultimately delivered between January 1988 and June 1989 with a gestational age > or = 26 weeks and who had an intrapartum amniotic fluid index composed the study group. The amniotic fluid index was determined by the four-quadrant technique on admission to labor and delivery. Oligohydramnios was defined as an amniotic fluid index < or = 5 cm (n = 170), borderline oligohydramnios as an amniotic fluid index 5.1 to 8.0 cm (n = 261), and normal amniotic fluid volume as an amniotic fluid index 8.1 to 20 cm (n = 336). Nine patients with an amniotic fluid index > 20 cm were excluded from data analysis. The oligohydramnios and borderline oligohydramnios groups were compared with the normal group with regard to antenatal, intrapartum, and postpartum variables. RESULTS: The groups had similar maternal age, parity, gestational age at delivery, and antenatal complications. Meconium-stained amniotic fluid occurred significantly less often in the oligohydramnios group compared with the normal group (relative risk 0.67, 95% confidence interval 0.49 to 0.92). However, variable decelerations occurred significantly more often in the oligohydramnios group compared with the normal group (relative risk 1.44, 95% confidence interval 1.12 to 1.87), and cesarean delivery for fetal distress also occurred significantly more often (relative risk 6.83, 95% confidence interval 1.55 to 30.4). There was no difference in Apgar scores or neonatal complications between groups. The efficacy of intrapartum-determined oligohydramnios predicting cesarean delivery for fetal distress gave a sensitivity of 78%, a specificity of 74%, a positive predictive value of 33%, and a negative predictive value of 95%. CONCLUSION: The amniotic fluid index for detecting intrapartum oligohydramnios is a valuable screening test for subsequent fetal distress requiring cesarean delivery.

Amniotic Fluid↗

Parathyroid hormone-related protein (PTHrP) concentrations in human amniotic fluid during gestation and at the time of labour.

To establish the changes associated with gestational age and labour status in parathyroid hormone-related protein (PTHrP) concentrations in the amniotic fluid, human amniotic fluid was collected from non-labouring and labouring women at < 37 weeks of gestation (preterm) and at term (> or = 37 weeks). PTHrP was assayed by a specific N-terminal radioimmunoassay. PTHrP concentrations in amniotic fluid obtained from non-labouring women were significantly lower at preterm (15-36 weeks; 14.1 +/- 2.5 pmol L(-1); n = 11) than at term (37-42 weeks; 39.3 +/- 7.6 pmol L(-1); n = 16; P < 0.0009). Concentrations of PTHrP in amniotic fluid obtained from labouring women were also significantly lower at preterm (27-36 week; 12.2 +/- 4.7 pmol L(-1); n = 4; P < 0.01) than at term (37-42 weeks; 63. 8 +/- 19.6 pmol L(-1); n = 9). There were no significant changes in concentration associated with labour status, either at preterm or at term. The physiological significance of elevated amniotic fluid concentrations of PTHrP has yet to be established, but the data are consistent with the suggestion that PTHrP plays a role in fetal membrane function during late gestation.

Amniotic Fluid↗

Correction of gamma-glutamyl transpeptidase deficiency in amniotic fluid of some cystic fibrosis fetuses by mixing with nondeficient fluids.

The deficiency of gamma-glutamyl transpeptidase activity, which was evident in some but not all cystic fibrosis amniotic fluids, could be corrected by mixing with either normal fluids or nondeficient cystic fibrosis fluids. Incubation of any amniotic fluid for 20 min at 62 degrees C resulted in total loss of gamma-glutamyl transpeptidase activity, but the activity could be restored by mixing with untreated nondeficient fluids. In contrast, no restoration could be obtained by mixing with untreated deficient cystic fibrosis fluids. Dialysis of amniotic fluids did not diminish their corrective capacity. Only the transpeptidation reaction was corrected and no correction was observed for the hydrolysis or autotranspeptidation of gamma-glutamyl p-nitroanilide in the absence of the glycylglycine or methionine acceptor. Plasma specimens did not have any corrective activity, although their gamma-glutamyl transpeptidase activity could be restored after heat inactivation by mixing with untreated nondeficient amniotic fluids. No correction was found for aminopeptidase or disaccharidase activities. These findings suggest that the deficient cystic fibrosis amniotic fluids probably contain normal quantities of the gamma-glutamyl transpeptidase enzyme but lack a heat-labile nondialyzable activator that is necessary for its transpeptidation catalytic performance. An assay for this transpeptidase activator may provide a valuable approach to identify at least a subgroup of cystic fibrosis patients.

Aminopeptidases↗

Human amniotic fluid mathematical model: determination and effect of intramembranous sodium flux.

OBJECTIVE: A recently described mathematical model of human amniotic fluid dynamics used known and estimated rates of fetal fluid production (lung liquid and urine) and composition (osmolality) to enable calculation of previously unmeasured routes of amniotic fluid resorption, including fetal swallowing and intramembranous (across the amnion) water flow. This "osmolar" model assumed that only free water resorption occurred across the intramembranous route. We hypothesized that intramembranous flow also may include solutes and electrolytes because significant concentration gradients exist between amniotic fluid and fetal plasma. We used mass balance analysis to determine the direction and magnitude of intramembranous sodium flux and to assess the ability of a newly described "sodium" model to predict changes in amniotic fluid volume in response to changes in intramembranous electrolyte flow. Mathematical modeling was used to predict changes in amniotic fluid volume in response to changes in intramembranous electrolyte flow. STUDY DESIGN: Model predictions were calculated using published values for human amniotic fluid and fetal urine composition and volume. Ovine studies were used to derive lung fluid volumes and composition. Fetal swallowing and intramembranous flow were independently determined using net amniotic fluid osmolar (osmolality model) and sodium (sodium model) balance. Differences between osmolality and sodium model predictions were normalized to calculate the net intramembranous sodium flux, assuming a net balance of intramembranous osmotic solute flow. RESULTS: Both sodium and osmolality models predicted swallowed volume to be greater than intramembranous flow until 28 to 32 weeks' gestation, after which the relationship reversed. However, the sodium model predicted greater intramembranous flow and lower swallowing rates compared with the osmolality model at all gestational ages. Osmolar mass balance required daily intramembranous sodium flux into the amniotic fluid, which increased with gestational age. Furthermore, assuming stable swallowing and intramembranous water flow, the model predicts that 5% increases or decreases in amniotic fluid solute concentrations caused by intramembranous flux result in polyhydramnios or oligohydramnios, respectively. CONCLUSION: Sodium and osmolality models demonstrate similarities in determinations of amniotic fluid dynamics. However, mass balance equations demonstrate a net intramembranous flow of sodium into the amniotic fluid under normal conditions. Mathematical modeling suggests that small alterations in daily intramembranous sodium flux may evoke large changes in amniotic fluid volume.

Amniotic Fluid↗

The sources and biochemical characteristics of cobalamin-binders in human amniotic fluid.

The sources and biochemical characteristics of cobalamin-binders in amniotic fluid were investigated. Using gel-permeation chromatography, cobalamin-binder, extracted from amniotic fluid at term, was recovered in a single peak with the molecular size of haptocorrin. Neonatal saliva also contained only haptocorrin. However, neonatal gastric juice contained two types of cobalamin-binders: haptocorrin and intrinsic factor. Amniotic fluid in midtrimester contained three types of cobalamin-binders: haptocorrin, intrinsic factor, and transcobalamin II. The cultured amnion cells secreted mainly apo-transcobalamin II. Concentrations of both apo-haptocorrin and salivary amylase in amniotic fluid increased as gestation advanced. These results suggest that cobalamin-binder in amniotic fluid in midtrimester originates from the fetal salivary gland, fetal gastric mucosa, and amnion cells, and that the contribution by the fetal salivary gland increases at term, when almost all cobalamin-binders in amniotic fluid are haptocorrin.

Amniotic Fluid↗

Amniotic fluid cortisol after premature rupture of membranes.

Total amniotic fluid cortisol concentration was studied in 12 patients at 24 to 34 weeks of gestation after premature rupture of membranes of one to eight days duration. Rupture of the membranes of less than 24 hours duration was associated with normal amniotic fluid cortisol concentrations. Rupture of membranes for more than 24 hours was associated in 10 of 12 patients with higher than normal amniotic fluid cortisol levels. The rise in amniotic fluid cortisol concentrations following premature rupture of the membranes may reflect increased fetal or maternal cortisol production or both, and could explain enhanced lung maturation in such infants.

Amniotic Fluid↗

Demonstration of tumor necrosis factor in human amniotic fluids and supernatants of placental and decidual tissues.

Human amniotic fluid samples obtained from normal pregnancies and supernatants of normal full-term placental and decidual tissues were assayed in a specific radioimmunoassay (RIA) and in a sensitive bioassay for tumor necrosis factor (TNF). TNF was detected in 91% of amniotic fluid samples and in all placental and decidual supernatants assayed by the RIA. Amniotic fluid samples collected during the second trimester of pregnancy contained significantly higher concentrations of TNF than samples collected during the third trimester, average concentrations being 2.5 ng/ml and 0.9 ng/ml, respectively. Parallel testing in the bioassay showed that most of the amniotic fluid TNF is biologically inactive, whereas placental and decidual supernatants had biologic activity correlating with TNF levels detected by the RIA. The quantities of TNF detected by the RIA ranged from 1.1 to 2.8 ng/ml in placental supernatants and from 3.9 to 8.5 ng/ml in decidual supernatants. The presence of TNF in normal amniotic fluids and in placental and decidual supernatants suggest that this cytokine has a physiologic role in human pregnancy.

Amniotic Fluid↗

Meconium-stained amniotic fluid.

The presence of meconium-stained amniotic fluid is a matter of importance when considering intrapartum management. Meconium-stained amniotic fluid is known to be associated with increased fetal and maternal risks, especially when fetal exposure has been extended and the fluid is concentrated. Selected primary and secondary sources are referenced in a summary that examines the implications and management of meconium-stained amniotic fluid for the newborn. The use of amnioinfusion in selected cases with oligohydramnios or meconium-stained fluid is discussed.

Amnion↗

Amniotic fluid volume reflects recent glycemic status in gestational diabetes mellitus.

OBJECTIVE: Our purpose was to determine the association between amniotic fluid volume and recent glucose status in gestational diabetes. STUDY DESIGN: Serial amniotic fluid index values, mean blood glucose levels, and percent hyperglycemia (> or = 120 mg/dl) 1 day and 1 week before the ultrasonographic examinations were prospectively collected in 399 gestational diabetics. Patients demonstrating at least one amniotic fluid index measurement within the normal range (i.e., 5 cm < amniotic fluid index < or = 20 cm) and at least one elevated measurement (i.e., amniotic fluid index > 20 cm) formed the study group. With each patient serving as her own control, glucose index values preceding normal and elevated amniotic fluid index values were compared. RESULTS: Significantly higher mean blood glucose values 1 day (114.7 mg/dl vs 102.8 mg/dl, p < 0.01) and 1 week before (111.0 mg/dl vs. 102.0 mg/dl, p < 0.05) were calculated for examinations resulting in elevated amniotic fluid index values compared with normal amniotic fluid index values, respectively. Similarly, significantly higher percents of hyperglycemia 1 day (32% vs 16.5%, p < 0.05) but not 1 week (30.8% vs 21.7%, p > 0.05) before the elevated amniotic fluid index were documented. CONCLUSION: Amniotic fluid volume reflects recent glycemic status in gestational diabetes mellitus.

Adult↗