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Study of the amniotic fluid from smokers and non-smokers in the Ames test.

Amniotic fluid from smokers and non-smokers was tested by the Salmonella/mammalian microsome test. Concentrated amniotic fluid from heavy smokers at term showed an increase in the number of revertants with increasing exposure to tar. However, some of the non-smokers had a higher number of revertants than the smokers. No significant differences were found between second-trimester samples from smokers and non-smokers, but the limited volumes available at this stage of pregnancy may be a source of error.

Adult↗

Intra-uterine growth retardation is associated with increased levels of magnesium in amniotic fluid.

Magnesium (Mg) and zinc (Zn) levels were measured in amniotic fluid of 30 normal pregnancies between the 16th and the 42nd week of amenorrhea and in 15 cases of intra-uterine growth retardation (IUGR) at 30-38th week of pregnancy. While Zn levels were stable throughout physiological pregnancy, the Mg concentration decreased progressively from 1.09 +/- 0.31 mg/100 ml (mean +/- SD) at the end of the second trimester to 0.75 +/- 0.20 (p less than 0.02) at term. In the case of IUGR, Mg levels (1.22 +/- 0.28) were higher than in controls (0.89 +/- 0.27, p less than 0.01) matched for gestational age. No differences in Zn levels were found between the two groups. Although the mechanisms leading to Mg accumulation in the amniotic fluid of growth-retarded fetuses remain unknown, these data support the role played by Mg in fetal growth.

Amniotic Fluid↗

Maternal plasma and amniotic fluid dehydroepiandrosterone-sulfate concentrations in preterm labor and delivery.

The purpose of this study was to determine whether preterm parturition is associated with changes in maternal plasma and amniotic fluid dehydroepian-drosterone-sulfate concentrations. A cross sectional study was constructed according to the gestational age at admission and response to tocolysis. Group 1 consisted of women admitted with preterm labor and intact membranes between 28 and 31 weeks and 6 days gestational age (n = 40). Group 2 included 40 patients with preterm labor between 32 and 36 weeks gestational age. Both groups were classified into two subgroups: preterm delivery within seven days of admission and term delivery. Commercially available immunoassay kits validated for amniotic fluid analysis of DHEA-S, were used to measure maternal plasma and amniotic fluid DHEA-S concentrations. Maternal plasma DHEA-S concentrations were significantly higher in women with preterm labor who delivered preterm than in those who delivered at term. (Group 1: median 800 ng/ml [range 100-1100] vs. median 200 ng/ml [70-800], P < 0.001; Group 2: median 850 ng/ml [300-1700] vs. median 300 ng/ml [90-1100], P < 0.001). In contrast, no significant differences were detected in amniotic fluid DHEA-S concentrations. Our data suggest that the rise in maternal plasma DHEA-S concentrations observed in patients with preterm labor may be related to the effects of stress during labor.

Adult↗

[Cadmium and lead concentrations in the amniotic fluid of pregnant smokers and non-smokers].

Many studies showed the unhealthy effect of smoking on the fetus. Several mechanisms for the unfavorable effect are discussed. In the last few years interest has been shown in the occurrence of heavy metals such as cadmium and lead in smoke. In animal experiments high doses of cadmium showed mutations, teratogenicity and low doses showed retardation of fetal growth. Amniotic fluid examinations of smoking and non-smoking pregnant women showed that there is no barrier to cadmium from the placenta. A direct correlation between cigarette smoking and cadmium concentration in the amniotic fluid was found. Cadmium was discussed as a reason for hypertension. But this was not verified in our study. There were no correlations between the cadmium and lead content of amniotic fluid and the gestational age, and disturbances of the pregnancy nor a correlation to the incidence of abortions. The lead concentration in the amniotic fluid of smokers and non-smokers showed no significant difference. This is attributed to the ubiquitous exposition to lead with increasing pollution of the environment. The relatively small additional amounts of lead in cigarettes do not increase the amount of lead in the amniotic fluid.

Abortion, Spontaneous↗

Amniotic fluid palmitic acid/stearic acid ratios. Lecithin/sphingomyelin ratios and palmitic acid concentrations in the assessment of fetal lung maturity in diabetic pregnancies.

The lecithin/sphingomyelin (L/S) ratio, palmitic acid concentration and palmitic to stearic acid (P/S) ratio were estimated on samples of amniotic fluid obtained from 66 patients with diabetes. These were compared with similar estimates on amniotic fluid obtained from 127 non-diabetic patients. At 35 to 40 weeks, significant differences were observed between the L/S ratio and palmitic acid concentration in diabetics and non-diabetics, whereas the P/S ratio was similar in the two groups. The amniotic fluid L/S ratio, palmitic acid concentration, and P/S ratio were estimated on amniotic fluid obtained from 20 diabetic patients within 48 hours of induction, and the clinical outcome of the newborn infant was used to assess the predictive value of the three parameters. In 19 out of 20 diabetics the P/S ratio correctly predicted fetal lung maturity, whereas the palmitic acid concentration was correct in 12 patients and the L/S ratio in only 10 patients.

Amniotic Fluid↗

Oral docosahexaenoic acid given to pregnant mice increases the amount of surfactant in lung and amniotic fluid in preterm fetuses.

OBJECTIVE: Our purpose was to determine whether docosahexaenoic acid increased surfactant production, as reflected by increased dipalmitoyl phosphatidylcholine, in mouse fetal lung and amniotic fluid. STUDY DESIGN: On day 9.5 of gestation, pregnant mice were given docosahexaenoic acid orally at 0, 5, 10, or 20 mg per day and were killed at day 16.5 (preterm) and day 19.5 (term) of gestation. Dipalmitoyl phosphatidylcholine was measured in fetal lung homogenates and amniotic fluid by high-performance thin-layer chromatography. RESULTS: Dipalmitoyl phosphatidylcholine values in lung were 0.22 +/- 0.27 microg/mg of total protein in preterm versus 1.96 +/- 0.57 microg/mg in term control fetuses. Pretreatment with 5, 10, or 20 mg docosahexaenoic acid increased dipalmitoyl phosphatidylcholine levels in preterm fetuses to 1.20 +/- 0.75, 1.60 +/- 0.67, and 3.28 +/- 0.44 microg/mg of protein, respectively. A similar trend was observed in amniotic fluid in which dipalmitoyl phosphatidylcholine levels were 1.86 +/- 3.70 microg/mL in preterm fetuses at baseline and increased to 7.81 +/- 1.21, 16.83 +/- 1.62 and 22.72 +/- 3.44 microg/mL after pretreatment for 7 days with 5, 10, and 20 mg docosahexaenoic acid (P<.05 compared to untreated mice). Dipalmitoyl phosphatidylcholine levels in amniotic fluid were 24.46 +/- 10.3 microg/mL in term control mice. CONCLUSION: The oral administration of docosahexaenoic acid to pregnant mice during pregnancy can induce dipalmitoyl phosphatidylcholine production and secretion, which is the major lipid component of surfactant.

Administration, Oral↗

Evaluation of gas-liquid chromatography for the rapid diagnosis of amniotic fluid infection: a preliminary report.

Gas-liquid chromatography has been proposed as a possible tool in the rapid diagnosis of amniotic fluid infections. The analysis is based on the identification of specific organic acids derived from bacterial metabolism when organisms are present within the amniotic fluid. We retrospectively subjected 69 samples of amniotic fluid which had been obtained by transabdominal amniocentesis to analysis by gas-liquid chromatography. Forty-seven samples were derived from patients who either were in premature labor or had premature rupture of membranes with associated premature labor. Twenty-two specimens which served as a comparison group were obtained from patients who underwent amniocentesis for assessment of fetal maturity. The results obtained from chromatographic analysis are presented, and the possible applications of this technique to the clinical situation are discussed.

Adult↗

Amniotic fluid inflammatory cytokines (interleukin-6, interleukin-1beta, and tumor necrosis factor-alpha), neonatal brain white matter lesions, and cerebral palsy.

OBJECTIVE: Ultrasonographically detectable neonatal brain white matter lesions are the most important identifiable risk factor for cerebral palsy. Inflammatory cytokines released during the course of intrauterine infections have been implicated in the genesis of brain white matter lesions and subsequent cerebral palsy. This study was undertaken to determine whether fetuses who subsequently were diagnosed to have periventricular brain white matter lesions could be identified by determining the concentrations of inflammatory cytokines in the amniotic fluid. STUDY DESIGN: Women with complicated preterm gestations underwent amniocentesis for clinical indications. Amniotic fluid concentrations of tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, and the natural interleukin-1 receptor antagonist were determined by immunoassay. Periventricular white matter lesions of the neonate were diagnosed by neurosonography. Univariate and multivariate analyses were conducted. RESULTS: Ninety-four women and their neonates were included in the study; white matter lesions were diagnosed in 24% (23/94) of the newborns. The mothers of newborns with brain white matter lesions had higher median concentrations of tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 (but not interleukin-1 receptor antagonist) in amniotic fluid than did those who were delivered of newborns without white matter lesions (p < 0.01 for each). Acute histologic chorioaminionitis was more common in the placentas of neonate with white matter lesions than in those without these lesions (82% [18/22] vs 42% [30/71], p < 0.005). Neonates with white matter lesions were delivered at a lower mean gestational age and birth weight and had a higher rate of significant complications (including respiratory distress syndrome, intraventricular hemorrhage, and infection-related complications) than did those without white matter lesions. The differences in median interleukin-1beta and interleukin-6 levels between these two groups remained significant after adjustment for gestational age and birth weight (interleukin-6: odds ratio 5.7, 95% confidence interval 1.3 to 24.4; interleukin-1beta: odds ratio 4.4, 95% confidence interval 1.1 to 17.0). Of the 94 newborns included in this study, 11 died before age 6 months and eight had cerebral palsy; all eight had white matter lesions and elevated cytokine levels in amniotic fluid. Histologic chorioamnionitis was more common in the placentas of neonates with cerebral palsy than in those without cerebral palsy (86% [6/7] vs 44% [33/75], p < 0.05). CONCLUSIONS: Infants at risk for development of brain white matter lesions can be identified by the concentrations of interleukin-6 and interleukin-1beta in amniotic fluid. Our findings support the hypothesis that inflammatory cytokines released during the course of intrauterine infection play a role in the genesis of brain white matter lesions.

Amniocentesis↗

Determination of phosphatidylglycerol in amniotic fluid for prediction of foetal lung maturity by microbore-column liquid chromatography.

Phosphatidylglycerol (PG) has been determined in amniotic fluid from 55 patients by using a microbore-column liquid chromatographic system. The present analysis time is 1 h 40 min for pretreatment of amniotic fluid and 20 min for a chromatographic run. From 2 ml of amniotic fluid, the PG content has been determined between 1.0 and 0.05 mg/dl. The detection limit of PG is 10 ng. As the injection volume (0.5 microliter) is small, repeated analyses are possible if necessary. It is concluded that, in the case of PG values over 0.10 mg/dl, a mother can deliver an infant without respiratory distress syndrome. This method is useful not only for the prenatal evaluation of lung maturity, but also for the assessment of any therapeutic effect.

Amniotic Fluid↗

Elevated second-trimester amniotic fluid interleukin-6 levels predict preterm delivery.

OBJECTIVE: Our purpose was to determine whether early second-trimester amniotic fluid interleukin-6 levels predict delivery before 34 weeks' gestation. STUDY DESIGN: We used stored second-trimester amniotic fluid samples obtained from women undergoing genetic amniocentesis from 1988 to 1996. Interleukin-6 levels were measured by enzyme-linked immunosorbent assay in samples from every case known to result in delivery from 20 to 34 weeks' gestation (n = 290), and 290 matched controls delivering at > or =37 weeks. Fetal aneuploidies, anomalies, and all cases delivering within 30 days of the amniocentesis (which were thought to be possibly procedure related) were excluded. RESULTS: Interleukin-6 levels were higher in cases than controls (1.9 +/- 5.2 vs 1.0 +/- 2.4 ng/ml, p = 0.004). Cases were grouped according to whether the preterm delivery was indicated or spontaneous: The mean interleukin-6 levels were significantly higher than controls in the spontaneous group (1.6 +/- 3.2 vs 0.8 +/- 1.2 ng/ml, p = 0.01) but not in the indicated group (1.4 +/- 4.0 vs 0.8 +/- 1.2 ng/ml, p = 0.12). In all samples the interleukin-6 level was negatively correlated with the gestational age at delivery (R = -0.11633, p = 0.007). CONCLUSION: Elevated early second-trimester amniotic fluid interleukin-6 levels are associated with preterm delivery, confirming that in some women this indicator of very early intrauterine inflammation predicts birth before 34 weeks' gestation.

Adult↗

Pro-EPIL forms are present in amniotic fluid and maternal serum during normal pregnancy.

Recent observations based on immunohistochemistry and RT-PCR demonstrate that early placenta insulin-like peptide (EPIL), encoded by the INSL4 gene, is present in the placenta during gestation. In the present study, we report on the development of a specific immunoassay, entirely based on two monoclonal anti-peptide antibodies (mAbs), and its use for the detection of pro-EPIL forms in biological fluids during pregnancy. One mAb directed against the C-connecting peptide was used to capture pro-EPIL forms and their binding was revealed by a radiolabeled anti-A chain mAb as the indicator. A composite synthetic peptide, encompassing the C- and A-domains, was utilized as the standard. Under these experimental conditions, the assay displays a sensitivity limit of 2 ng/mL. Pro-EPIL molecular forms were detected in both amniotic fluid and maternal serum of pregnant women. At 12 to 16 weeks of pregnancy, the pro-EPIL level was higher in amniotic fluid (246 +/- 50.8 ng/mL) than in maternal serum (5 +/- 2.0 ng/mL). As gestation advanced, so the concentration of pro-EPIL forms decreased in amniotic fluid while its level increased in maternal serum. Interestingly, in amniotic fluid, the pattern of pro-EPIL concentration during pregnancy is very similar to that observed for human chorionic gonadotropin (hCG) and its free subunits. The pattern of serum pro-EPIL concentration is similar to that of the free alpha-subunit. Together with our previous immunohistochemical observations, these results indicate that pro-EPIL is preferentially secreted by cytotrophoblasts in amniotic fluid and that the biosynthesis of hCG subunits and EPIL may be regulated by common pathways. Overall, our observations strongly suggest that EPIL may play a critical physiological role during embryonic and foetal development.

Adult↗

Characterization of a type IV procollagen synthesized by human amniotic fluid cells in culture.

Fetal epithelioid cells, isolated from human amniotic fluid, synthesize and secrete a type IV-like procollagen characterized by a unique pattern of cyanogen bromide (CNBr)-produced peptides. The procollagen is disulfide-bonded and, after reduction, migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a doublet between collagen beta components and pro-alpha 1(I) chains. No conversion of the procollagen to collagen or to procollagen intermediates is observed in cell culture. The procollagen was purified by salt fractionation and ion exchange chromatography; its amino acid composition resembles that of collagenous proteins extracted from basement membranes, with a high 3- and 4-hydroxyproline and hydroxylysine content and low levels of alanine and arginine. The major products obtained after limited proteolytic digestion of the protein retain interchain disulfide bonds and, after reduction, migrate on sodium dodecyl sulfate-polyacrylamide gel electrophoresis near intact pro-alpha 1(I) chains. The procollagen is secreted efficiently by amniotic fluid cells despite almost complete inhibition of peptidyl hydroxylation but, unlike type I procollagen, the secreted underhydroxylated chains lack interchain disulfide bonds. Since these cells also secrete fibronectin and elaborate an extensive extracellular matrix, the system should prove useful in the study of cell-matrix interactions.

Amino Acids↗

[Congenital human cytomegalovirus infection: value of human cytomegalovirus DNA quantification in amniotic fluid].

A quantitative PCR assay (RS Elosa CMV, Lambdatech) was used to quantitate HCMV DNA in maternal amniotic fluid of 12 fetuses with congenital infection (group 1) and of 10 fetuses without congenital infection (group 2). HCMV detection was performed for both groups using culture and qualitative PCR. Histologic examinations of fetal tissues and placenta were carried out for 9 patients from group 1. The amniotic fluid viral loads were negative in all patients of group 2. In group 1, all viral loads were high (from 1.105 to > 107 cop/mL) and no difference was observed between symptomatic and asymptomatic foetuses. Further evaluation on larger samples is needed to define more precisely the pronostic value of HCMV DNA quantification in amniotic fluid.

Amniotic Fluid↗

Amniotic fluid beta-endorphin: a prognostic marker for gastroschisis?

PURPOSE: The aim of this work was to study amniotic fluid beta-endorphin as a potential predictor for postnatal morbidity in gastroschisis. METHODS: Beta-endorphin was assayed in 43 amniotic fluid samples from 13 pregnant women with fetal gastroschisis undergoing diagnostic amniocentesis or therapeutic amnioinfusion and compared with 33 controls. Within the gastroschisis group, the authors investigated the relationship between postnatal morbidity and the peak value of amniotic fluid beta-endorphin (AFBE). RESULTS: Ten AFBE values in 6 cases of gastroschisis were above the upper limit of the 95% confidence interval derived from controls. Postnatal morbidity was significantly higher when peak AFBE exceeded 10 microg/L (n = 4 pregnancies) compared with below 5 microg/L (n = 9 pregnancies), as shown by mean duration of mechanical ventilation (15.2 v 3 days; P =.01), of parenteral feeding (77 v. 18.7 days; P =.04), and of hospitalization (84 v 32.2 days; P =.04). There was no statistically significant association between postnatal morbidity markers and prenatal dilation of fetal bowel. CONCLUSIONS: The most severe cases of gastroschisis are associated with high levels of AFBE. The authors speculate that this fetal hormonal response could result from stress or pain caused by prenatal bowel damage.

Amniocentesis↗

Amniotic fluid prealbumin as a potential marker of fetal abnormalities.

Normal amniotic fluid prealbumin (PAB) levels between 16 and 41 weeks of pregnancy are demonstrated in 175 samples from various gestational ages. PAB was measured quantitatively by immunonephelometry and qualitatively estimated by electrophoresis while total protein content was measured by colorimetry. The percent of PAB values in the total protein content is inversely correlated to the gestational age. Fetal abnormalities and abnormal conditions in pregnancy diagnosed in the second trimester and early third trimester (16-32 weeks) show significantly (p < 0.01) higher than expected PAB values (25.9 +/- 4.6 mg/l, range 19.5-35.6, n = 23) in comparison to normal fetuses and normal pregnancies of the same age (13.6 +/- 3.6 mg/l, range 4.3-19.8, n = 87). The remaining 65 samples of normal third trimester amniotic fluid were evaluated in comparison to the second trimester normal values, and inverse correlation in respect to advancing pregnancy was observed. PAB values were corrected for the total protein content and the resulting ratio had the lowest variation compared to PAB and total protein values. It is proposed that PAB, although not specific for a particular disorder, expressed as a ratio of the total protein content is a potential biochemical marker for certain fetal defects and complications of pregnancy.

Amniotic Fluid↗

Amniotic fluid optical density: relationship to L:S ratio, phospholipid content, and desquamation of fetal cells.

The relationship between amniotic fluid optical density at 650 nm (OD650) and the lecithin:sphingomyelin (L:S) ratio was compared in 174 samples of amniotic fluid from 143 patients. An OD650 greater than 0.15 accurately predicted and L:S ratio greater than 2.0 with an overall reliability of 67.9%. There were 19.5% false-negative and 12.6% false-positive predictions. The OD650 was found to have close correlation with the number of fetal squames in suspension in amniotic fluid. Pure surfactant phospholipid standards were found to exhibit negligible absorbance at 650 mm. OD650 may be more closely related to fetal cutaneous maturity than to pulmonary surfactant content and bears only an indirect relationship with pulmonary preparedness for independent existence. The OD650 test does not appear to be sufficiently reliable and specific to replace phospholipid determination as a data phase upon which to make clinical decisions regarding fetal pulmonary surfactant maturity.

Amniotic Fluid↗

Antibacterial activity in human amniotic fluid: relationship to zinc and phosphate.

Antibacterial activity, tested with B. subtilis, was present in all 44 human amniotic fluids at between 36 and 42 weeks of pregnancy and was unrelated to gestational age within these limits, or to maternal age or parity. High-molecular-weight (beta-lysin) activity, present in all samples, was directly related to concentration of bound zinc, but not to that of free zinc. Low-molecular-weight antibacterial activity (less than 1000 Daltons) was only present in nine specimens, but sub-threshold concentrations were demonstrable by concentrating the ultrafiltrates in more than half of the remaining specimens. Low-molecular-weight activity in the ultrafiltrates was directly related to the concentration of bound zinc in the original amniotic fluid. Neither high- nor low-molecular-weight antibacterial activities were associated with amniotic fluid phosphate levels.

Adult↗

Efficacy of bulb versus DeLee suction at deliveries complicated by meconium stained amniotic fluid.

A combined obstetric-pediatric approach to the management of deliveries complicated by meconium-stained amniotic fluid has been recommended to decrease the incidence of meconium aspiration syndrome. Although pharyngeal suction with a DeLee catheter is generally recommended, many obstetricians prefer to use a bulb syringe for suction at the perineum. A thorough review of the literature reveals no prospective studies in humans comparing the two techniques. In the present study, these two techniques of pharyngeal suction were prospectively compared at deliveries complicated by meconium-stained amniotic fluid. Of the total 2874 deliveries, 127 (4%) were complicated by moderate or thickly meconium-stained amniotic fluid, and 107 of these were included in the study. There were 53 infants in the DeLee-suctioned group and 54 in the bulb-suctioned group. Of the 107 studied infants, four (4%) developed meconium aspiration syndrome, three in the DeLee group and one in the bulb group. There was no significant difference in the amount of meconium found below the vocal cords, comparing pharyngeal DeLee suction to bulb suction (0.22 cc versus 0.24 cc; p = NS). In conclusion, the data fail to support a significant difference in efficacy between bulb and DeLee suction in clearing the naso- and oropharynx of the neonate of meconium in cephalic-presenting vaginal or cesarean delivery.

Adult↗

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