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Isolation and purification of pulmonary surfactant-associated protein A from human amniotic fluid and its bioactivity assessment.

OBJECTIVE: To isolate and purify pulmonary surfactant-associated protein A(SP-A) from human amniotic fluid. METHODS: Human amniotic fluid was collected from 11 parturients and SP-A was isolated by extraction and dialysis followed by purification procedures. The specificity of SP-A was detected by Western blotting and the bioactivity assessed by MTP-1 film balance. RESULTS: An amount of 12 mg (20 ml) purified SP-A was obtained, and the protein presented a single band upon SDS-PAGE with relative molecular mass of 31 000. Western blotting also displayed one specific band. Film balance demonstrated that the minimum surface tension alphamin of Exosurf, an synthetic pulmonary surfactant, was 7.7+/-0.3, significantly higher than alphamin of Exosurf and SP-A mixture(1.5+/-0.22, P<0.05), indicating enhanced bioactivity of the mixture. CONCLUSION: Dialysis can effectively purify SP-A from human amniotic fluid with little damage of its biological activity.

Journal Article↗

A study of the procoagulant properties of amniotic fluid and their correlation with the lecithin/sphingomyelin ratio.

The procoagulant properties of amniotic fluid have been studied in samples taken throughout pregnancy and the results correlated with the lecithin/sphingomyelin ratio: acceleration effects on the prothrombin time. Russell's viper venom test, activated partial thromboplastin time and specific factor VII and X assays and contact acceleration. Although amniotic fluid is shown to function as a complete thromboplastin and to activate factor X directly, none of these tests of procoagulant activity of amniotic fluid is a reliable index of fetal lung maturity.

Amniotic Fluid↗

Zinc concentration in gravida's serum and amniotic fluid during normal pregnancy.

Zinc concentration was determined in maternal serum and amniotic fluid during normal pregnancy. The nonpregnant zinc concentration in the serum of about 110 micrograms% decreases gradually during pregnancy to about half the normal level. Amniotic fluid zinc concentration shows a slight decrease and remains nearly constant until the 34th to 36th week of gestation, and then increases sharply (statistically significant) reaching the highest level at 40-42 weeks from the last menstrual period. The maternal-fetal metabolism of zinc, the various factors effecting zinc levels in maternal serum and amniotic fluid during pregnancy have been discussed in view of our results. These may serve as data base for further studies on the role of zinc in normal and pathological human pregnancies.

Amniotic Fluid↗

Changes in amniotic fluid glucose, beta-hydroxy-butyrate, glycerol, and lactate concentration in diabetic pregnancy.

Amniotic fluid glucose, beta OH butyrate, glycerol, and lactate concentrations were measured in 75 samples collected in the third trimester of pregnancy from 50 diabetic patients, all but four of whom required insulin. Increases in maternal fasting plasma sugar were accompanied by corresponding increases in amniotic fluid glucose and on occasion increases in amniotic fluid beta OH butyrate. These data correspond to previous reports of placental glucose transfer and in addition, provide statistically significant evidence of placental betaOH butyrate transfer since the hyperglycemic, hyperinsulinemic fetus of a diabetic mother would be a poor primary source for ketogenesis. Relatively poor correlation of elevated fluid levels of these solutes to fetal outcome probably reflects a low incidence of maternal hyperglycemia, ketogenesis. Relatively poor correlation of elevated fluid levels of these solutes to fetal outcome probably reflects a low incidence of maternal hyperglycemia, ketoacidosis, and over-all reduced neonatal morbidity-mortality rates in this group of metabolically well-controlled, predominantly insulin-requiring diabetic patients managed in a regional high-risk perinatal center.

Adult↗

Relationships among maternal and fetal amino acid blood concentrations and amniotic fluid amino acid levels in pregnant rhesus monkeys.

This study was designed to explore relationships among maternal and fetal amino acid blood concentrations and amniotic fluid amino acid levels in pregnant rhesus monkeys. Although there was no constant pattern for all amino acids in the three body fluids, we observed specific patterns for most neutral amino acids, imino acids, and acidic amino acids relative to their concentrations in maternal blood, fetal blood, and amniotic fluid. Moreover, there was no correlation between fetal phenylalanine blood levels and phenylalanine concentrations in amniotic fluid indicating that amniotic fluid phenylalanine levels would not be useful to monitor fetal phenylalanine blood concentrations during pregnancy.

Amino Acids↗

Proliferative stimulation of lymphocytes by calmodulin binding proteins isolated from amniotic fluid.

In view of the role of calmodulin and calmodulin binding proteins in modulating the second messenger functions of Ca2+, we studied the presence of such proteins in amniotic fluid, which may be considered as a dynamic medium for promoting foetal growth. Affinity chromatography of amniotic fluid proteins revealed the presence of calmodulin binding proteins in samples obtained either at 28 or 36 wk of pregnancy. The relative content of these proteins increased in amniotic fluid from 1.5 mg/g total protein at 28 wk to 3.6 mg/g at full term of pregnancy. Culturing murine splenocytes in presence of the isolated calmodulin binding proteins (10 micrograms protein/10(6) cells) resulted in nearly 4-fold enhancement of 3H-thymidine incorporation into them as compared to controls. In comparison, similar incorporation of the radiolabel into lymphocytes obtained from cord blood was enhanced only by 2 fold in presence of calmodulin binding proteins, though at a much lower protein concentration (50 ng/10(6) cells). SDS-PAGE on 12.5 per cent gels of eluates obtained from calmodulin-agarose columns, followed by overlay of corresponding western blots with biotinylated calmodulin revealed the presence of a 68 kDa calmodulin binding protein in samples collected either at 28 wk or at full-term of pregnancy. In addition, the amniotic fluid also contained 83 kDa calmodulin binding protein at 28 wk. This first-time demonstration of mitogenic, calmodulin binding proteins in amniotic fluid suggests that such mitogens may participate in promoting foetal growth.

Amniotic Fluid↗

Mifepristone (RU 486) and its incidence on amniotic fluid volume in near term fetal macaque monkeys.

OBJECTIVE: To evaluate a possible effect of Mifepristone on amniotic fluid volume in near term pregnant macaque monkeys. STUDY DESIGN: Six pregnant near term monkeys (144 +/- 11 days of gestation) were either treated intramuscularly by Mifepristone (10 mg/kg, n = 3) or a placebo (n = 3). Amniotic fluid volumes were measured by a dilution technique using blue Evans, just prior to the injection of Mifepristone or placebo (t0h) and 24 h later (t24h). RESULTS: Amniotic fluid volume (AFV) measured at t0h was found to be statistically the same in placebo and Mifepristone-treated macaque monkeys (95 +/- 31 vs. 126 +/- 25 ml, respectively, P > 0.05). AFV was found to be lower at t24h when compared to AFV at t0h in placebo-treated (88 +/- 31 vs. 95 +/- 31 ml, respectively, P < 0.05) and in Mifepristone-treated macaque monkeys (72 +/- 7 vs. 126 +/- 25 ml, respectively, P < 0.05). Moreover, the ratio of AFV (t24h/t0h) was found to be significantly lower in Mifepristone-treated monkeys when compared to the placebo group (57.9 +/- 5.7% vs. 92.2 +/- 2.1%, P < 0.05) indicating a specific effect of the antiprogestin on AFV. CONCLUSION: In near term macaque monkeys, Mifepristone at 10 mg/kg induces a significant decrease of amniotic fluid volume within 24 h.

Amniotic Fluid↗

[Correlation of cotinine level in amniotic fluid, umbilical artery blood and maternal blood].

The aim of the present study was to determine the correlation of cotinine levels in amniotic fluid and in fetal and corresponding maternal blood samples. Amniotic fluid samples (N = 130) were taken during second trimester amniocentesis, umbilical artery blood samples (N = 75) at birth, both together with corresponding maternal blood samples. Self-reported smokers showed maternal serum cotinine levels > 15 ng/ml in 93%, self-reported nonsmokers levels < 15 ng/ml in 89%. Correlation of corresponding values for cotinine was 0.81-0.92. Cotinine values were increased in fetal blood and amniotic fluid in comparison to maternal serum levels. Despite the fact that pathophysiology is not fully understood, an accumulation of nicotine and its metabolites both in the fetus and in the amniotic fluid appears to be evident.

Amniocentesis↗

Disposition of ethanol in maternal venous blood and the amniotic fluid.

The disposition of ethanol and acetaldehyde in maternal venous blood and amniotic fluid was studied over a time course following ingestion of 0.3 g/kg of ethanol by six healthy pregnant women at 16- to 18-weeks gestation. A time lag occurred in appearance and elimination of ethanol in and from the amniotic fluid when compared with maternal venous blood. Acetaldehyde was present in the maternal venous blood of four subjects but was found in the amniotic fluid in only one subject. Data from this clinical trial can be used by nurses who counsel pregnant women.

Acetaldehyde↗

[The transfer of drugs into the amniotic fluid].

On the basis of the present literature and our own results a review will be given about the importance of the physico-chemical properties of drugs and of different factors of biological system "mother, fetus and amniotic fluid" for the passage of drugs into amniotic fluid. The amniotic fluid is relieved as a possible distribution volume for drugs.

Amniotic Fluid↗

Estimation of surfactant lecithin in amniotic fluid and prediction of respiratory distress syndrome.

The quantitative and qualitative analyses of lecithin in the human amniotic fluid in relation to the antenatal prediction of developing respiratory distress syndrome were described. Surfactant lecithin (disaturated and/or dipalmitoyl lecithin) was estimated for the first time in 42 amniotic fluid specimens of 21-42 weeks gestation. Surfactant lecithin was present in considerable amounts in specimens obtained near or at term except in the ones associated with infants who subsequently developed respiratory distress syndrome. One infant developed typical respiratory distress syndrome despite the normal total lecithin concentration of amniotic fluid at birth. The fatty acid analysis, however, revealed that the surfactant lecithin concentration was extremely low, showing a great discrepancy between total lecithin and surfactant lecithin concentrations. Thus the estimation of disaturated and/or dipalmitoyl lecithin in the amniotic fluid seems to be the most precise sign of a potential risk of respiratory distress syndrome.

Amniotic Fluid↗

N-terminal peptide of pro-opiomelanocortin in human amniotic fluid.

N-terminal peptide of pro-opiomelanocortin (N-POMC) was measured in the human amniotic fluid. At the gestational age of 16 to 20 weeks, the radioimmunoassay with three different antibodies demonstrated the respective values of 2.39 +/- 0.78, 4.69 +/- 2.27, and 5.92 +/- 2.66 ng/ml. These values are approximately 10 times higher than the measurements in the plasma of women at the corresponding gestational period. The amniotic fluid collected during the delivery had significantly lower concentrations of N-POMC than the amniotic fluid at 16 to 20 weeks' gestation. However, the plasma values of N-POMC had increased approximately three times when measured at delivery and compared with the plasma values at 16 to 20 weeks' gestation. Adrenocorticotropic hormone, measured simultaneously with N-POMC in some of the samples, showed changes similar to those in N-POMC. The N-POMC immunoreactivity from the amniotic fluid has the same retention time on reversed-phase high-performance liquid chromatographic separation as the peptide purified from the human pituitary gland, thus indicating the identity of both peptides.

Adrenocorticotropic Hormone↗

Predictive power of maternal serum and amniotic fluid CRP and PAPP-A concentrations at the time of genetic amniocentesis for the preterm delivery.

OBJECTIVE: To investigate whether maternal serum and amniotic fluid CRP and PAPP-A concentrations at the time of genetic amniocentesis are markers of preterm delivery. STUDY DESIGN: One hundred and forty-one pregnant women were included in this prospective study. Amniotic fluid and maternal serum CRP and PAPP-A concentrations were determined by using commercially available kits. Receiver-operating characteristic (ROC) analysis was performed to determine the efficacy of maternal serum and amniotic fluid CRP and PAPP-A levels in predicting women with preterm delivery. RESULTS: The prevalence of spontaneous preterm delivery before 37 weeks of gestation was 9.9%. ROC analysis revealed that amniotic fluid CRP level was the only parameter, which had a significant power in the prediction of preterm delivery. The optimum cut-off level was 0.65 mg/L. The sensitivity and specificity were 92.9% and 78.7%, respectively. CONCLUSION: The amniotic fluid CRP level has a high sensitivity and specificity in the prediction of preterm delivery and this may be helpful in predicting preterm delivery during genetic amniocentesis.

Adult↗

Elevated amniotic fluid C-reactive protein at the time of genetic amniocentesis is a marker for preterm delivery.

OBJECTIVE: A pre-existing intrauterine inflammation in the first half of gestation has been proposed as a possible condition that leads to preterm delivery. Indeed, elevated levels of inflammatory mediators (eg, interleukin-6, tumor necrosis factor) in midtrimester amniotic fluid have been found in cases of preterm delivery and/or spontaneous abortion. The objective of this study was to investigate whether the amniotic fluid C-reactive protein level at the time of genetic amniocentesis is a marker for spontaneous preterm delivery before 34 and 37 weeks of gestation. STUDY DESIGN: Women who underwent genetic amniocentesis between 15 and 18 weeks of gestation with (1) singleton gestation, (2) uneventful pregnancy course before the amniocentesis, and (3) absence of fetal abnormalities were included in the study. Patients with abnormal karyotype were excluded. C-reactive protein concentration was measured in amniotic fluid and in maternal blood immediately after genetic amniocentesis. All patients were followed until delivery for the occurrence of pregnancy complications. Nonparametric tests and receiver-operating characteristic curve analysis were used for statistical purposes. RESULTS: The prevalence of spontaneous preterm delivery before 34 and 37 weeks was 3.3% (10 of 306 pregnancies) and 8.5% (26 of 306 pregnancies), respectively. Women with preterm delivery at <37 weeks had a higher median (range) of amniotic fluid C-reactive protein concentration than those women who delivered at term (median, 113.3 ng/mL [range, 16-623 ng/mL] vs median, 57.8 ng/mL [range, 0-808.9 ng/mL]; P <.005). Women with preterm delivery at <34 weeks had a higher median (range) amniotic fluid C-reactive protein concentration than those women who delivered at term (median, 183.8 ng/mL [range, 46.5-447 ng/mL] vs median, 57.8 ng/mL [range, 0-808.9 ng/mL]; P <.005]. No correlation was found between amniotic fluid C-reactive protein and maternal blood C-reactive protein concentrations. No relationship was found between maternal blood C-reactive protein concentration and preterm delivery before either 34 or 37 weeks. Amniotic fluid C-reactive protein concentration of >110 ng/mL had a sensitivity of 80.8% and a specificity of 69.5% in the prediction of spontaneous preterm delivery at <34 weeks. CONCLUSION: This study supports the theory that a subclinical intrauterine/fetal inflammatory process early in gestation may be important for the occurrence of preterm delivery in the second half of gestation.

Adult↗

Expectant management of premature rupture of membranes with amniotic fluid cultures positive for Ureaplasma urealyticum alone.

OBJECTIVE: Our purpose was to determine maternal and neonatal outcome in patients with premature rupture of membranes and amniotic fluid cultures positive for Ureaplasma urealyticum alone. STUDY DESIGN: Amniocentesis was performed on 225 patients with premature rupture of membranes at < or = 34 completed weeks of gestation. Amniotic fluid was cultured for aerobic and anaerobic bacteria, Mycoplasma hominis, and Ureaplasma urealyticum. Patients with cultures positive for Ureaplasma urealyticum alone were managed expectantly, including erythromycin and daily biophysical profile examinations. RESULTS: Thirty-three patients (15%) had amniotic fluid cultures positive for Ureaplasma urealyticum alone, and clinical chorioamnionitis developed in three of these patients. There were no suspected or documented cases of neonatal sepsis. The incidence of infectious morbidity in patients with amniotic fluid cultures positive for Ureaplasma urealyticum alone was not significantly different from that seen in patients with negative cultures. CONCLUSION: An amniotic fluid culture positive for Ureaplasma urealyticum alone may not be an indication for a preterm delivery in patients with premature rupture of membranes.

Amniotic Fluid↗

Interleukin-6 and interleukin-8 in cervical and amniotic fluid: relationship to microbial invasion of the chorioamniotic membranes.

OBJECTIVE: To evaluate the levels of interleukin (IL)-6 and IL-8 in cervical and amniotic fluid in relation to the presence of bacteria in the membranes in women in preterm labour (PTL). DESIGN: A prospective follow up study. SETTING: Sahlgrenska University Hospital, Göteborg, Sweden. Sample Women with singleton pregnancies (<34 weeks) presenting with PTL (n = 30). METHODS: Amniotic fluid was retrieved transabdominally and cervical fluid was sampled from the uterine cervix at admission and analysed for IL-6 and IL-8 with enzyme-linked immunosorbent assay (ELISA). At birth, the chorioamniotic membranes were separated and samples for polymerase chain reaction (PCR) for Ureaplasma urealyticum and Mycoplasma hominis and general culture were obtained. MAIN OUTCOME MEASURE: IL-6 and IL-8 in relation to microbial invasion of the chorioamniotic membranes. RESULTS: Bacteria were found in the membranes in 8 of 21 patients in PTL for whom chorioamnion as well as amniotic fluid PCR and cultures were available. Cervical IL-6 was associated with detectable bacteria in the chorioamniotic membranes in women in PTL (median 8.2 ng/mL vs 0.73 ng/mL; P = 0.01). The IL-6 (median 13 ng/mL vs 1.7 ng/mL; P = 0.004) and IL-8 (median 7.2 ng/mL vs 0.28 ng/mL; P = 0.01) levels in amniotic fluid were higher in PTL cases in which bacteria were found in the chorioamniotic membranes. CONCLUSION: IL-6 in cervical fluid and IL-6 and IL-8 in amniotic fluid were higher in those PTL cases in which bacteria were found in the chorioamniotic membranes.

Adult↗

The production mechanism of amniotic fluid dopamine in rats.

The production of dopamine in amniotic fluid was studied using Wistar rats. The dopamine concentrations in the amniotic fluid increased significantly between each day of gestation from day 18 to day 21, and a marked increase was observed during the last two days. The concentrations of dopamine in the fetal plasma, however, did not change with the gestational age. There was a significant increase in the fetal plasma L-dopa, which is the precursor of dopamine, without any changes in maternal plasma L-dopa levels during late gestation. Dopa decarboxylase (DDC) activity was also measured in the fetal rat kidney from gestation day 17 to day 21. DDC activity, which was low on day 17, increased with advanced gestation. DDC activity on day 21 was 0.68 nmol/mg tissue/h, about 13-fold higher than on day 17. These results suggest that fetal plasma L-dopa may be converted to dopamine by fetal kidney DDC, and that the dopamine is voided into the amniotic cavity in fetal urine.

Amniotic Fluid↗

The effects of partial and total amniotic fluid exclusion on excisional fetal rabbit wounds.

Previous studies have consistently demonstrated that fetal rabbit excisional wounds exposed to amniotic fluid do not heal in utero. This study examines fetal wounds covered to exclude amniotic fluid. Full-thickness excisional wounds were created on fetal rabbits on day 25 of gestation. The following three treatment groups were examined: uncovered (n = 19 live); covered, a silicone cover was sutured over the wound (n = 14 live); and donut, a silicone cover containing a 2-mm central hole was sutured over the wound (n = 6 live). Fetuses were returned to the uterus and harvested by cesarean section 72 hr postoperatively. Fetal (covered) wounds excluded from amniotic fluid contracted and reepithelialized up to confluence. Sixteen of 19 uncovered wounds expanded, 3 were unchanged; minimal reepithelialization was present in all wounds. Partially (donut) covered wounds exhibited an intermediate response. This study shows that excisional wounds in the rabbit fetus respond to injury by contracting and reepithelializing when excluded from amniotic fluid.

Amniotic Fluid↗