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Epidermal growth factor in urine of pregnant women and in amniotic fluid throughout pregnancy.

To investigate the role of epidermal growth factor (EGF) in feto-placental development, we measured the urinary and amniotic fluid EGF levels throughout pregnancy. Thirty urinary samples of non-pregnant women, 85 of normal pregnant women, 21 of women with toxemic pregnancy, 17 of postpartum women and 30 of newborns, and 55 amniotic fluid samples of pregnant women with a variety of conditions necessitating amniotomy and amniocentesis at 25-39 weeks of gestation were collected. EGF concentrations were measured by double-antibody radioimmunoassay. Urinary EGF levels of pregnant women reached their peak (24.6 +/- 6.7 ng/mg creatinine) at 19-22 gestational weeks; after that, they slightly decreased. Although there is no significant difference between the urinary EGF levels of non-pregnant women (19.0 +/- 5.1) and those of pregnant women (18.1 +/- 3.2), the EGF levels of toxemic women (12.2 +/- 1.5) were lower than those of normal pregnant women. The levels in puerperium women were similar to those found during pregnancy. However, the neonates had higher urinary EGF concentrations than those in pregnant women. On the other hand, EGF levels in amniotic fluid were higher according to gestational weeks and the levels of intrauterine growth retardation (IUGR) cases lower compared with normal pregnancy. Furthermore, EGF concentrations in amniotic fluid have a significant correlation with the creatinine levels in amniotic fluid. These data suggest that EGF plays an important role in fetoplacental development and it is possible that the measurement of amniotic fluid EGF might become available for the clinical assessment of fetal maturation.

Amniotic Fluid↗

Amniotic fluid and fetal tissues are not heated by obstetric ultrasound scanning.

Amniotic fluid temperature in first trimester pregnancies and fetal subcutaneous tissue and amniotic fluid temperature in second trimester pregnancies, measured by a thermocouple probe, did not increase during sector, linear-array or Doppler ultrasound scanning. The mean fetal muscle temperature was higher (36.9 degrees C) than mean amniotic fluid temperature (36.6 degrees C) during the second trimester.

Amniotic Fluid↗

Is induction of fetal diuresis with intraamniotic furosemide effective for the removal of intestinal waste products from amniotic fluid?

AIM: In gastroschisis, contact with amniotic fluid (AF) causes intestinal damage. Intraamniotic meconium has been shown to be responsible for the intestinal damage, and intestinal damage has been shown to correlate with intraamniotic meconium concentrations. Intraamniotic meconium below a threshold level does not cause intestinal damage. Intraamniotic meconium concentrations can be lowered by AF exchange. Can induction of foetal diuresis by an intraamniotic injection of furosemide be used as an alternative method for the same purpose? METHOD: Pregnant rabbits on the 23rd - 25th gestational days (normal gestation time: 31 - 33 days) were divided into two groups, the control group and the furosemide group. Initial AF samples were taken, then either 5 mg/kg furosemide or a placebo was injected into the amniotic cavity. Final AF samples were obtained 6 hours later. AF urea nitrogen, creatinine, amylase, alkaline phosphatase and bilirubin levels were determined. RESULTS: There was no significant difference between the initial and final levels of AF urea nitrogen, creatinine, bilirubin, amylase, and alkaline phosphatase in the control group, while the final AF urea nitrogen and creatinine levels of the furosemide group were not significantly different from the initial levels (p > 0.05). Final AF bilirubin, amylase and alkaline phosphatase levels of the furosemide group were significantly decreased compared with initial levels (p < 0.01). CONCLUSION: Induction of foetal diuresis with intraamniotic furosemide is effective for the removal of intestinal waste products from amniotic fluid.

Amnion↗

Infection and labor: the detection of endotoxin in amniotic fluid.

Intra-amniotic infections are frequently caused by gram-negative organisms. Lipopolysaccharide, or endotoxin, is a component of the cell wall of these bacteria. The Limulus amebocyte lysate assay is the most sensitive test clinically available for the detection of endotoxin in biologic samples. The diagnostic value of the Limulus amebocyte lysate assay test for the detection of intra-amniotic infection has been examined and compared with that of the Gram stain. The Gram stain had a 60.8% sensitivity and 9.6% false positive rate. The Limulus amebocyte lysate assay had a 69% sensitivity and a 4.8% false positive rate. When both tests were used in combination, the sensitivity improved to 95.6%, whereas the false positive rate was 14.3%. The Limulus amebocyte lysate assay test is a rapid, sensitive, inexpensive, and simple test for the detection of gram-negative intra-amniotic infections.

Amniotic Fluid↗

The effect of filtration of amniotic fluid on the growth of Chlamydia trachomatis and Escherichia coli.

There is increasing concern about Chlamydia trachomatis infection during pregnancy, because of reports of increased maternal, fetal, and neonatal risks. Amniotic fluid is known to possess antibacterial activity and has recently been shown to inhibit formation of chlamydial inclusions in McCoy cell culture. To further characterize the anti-chlamydial factor, we investigated the effect of filtering the fluid (0.45 microns pores) prior to incubation. Amniotic fluid was obtained from 12 women at term gestation, either by amniocentesis, or at cesarean section, Chlamydial inclusion formation was studied in McCoy cell cultures, and Escherichia coli growth was studied by a plate-count method. Filtered amniotic fluid had significantly less inhibitory activity against chlamydial inclusion formation than nonfiltered fluid did. Both filtered and nonfiltered amniotic fluid were equally effective in inhibiting E. coli colony growth. These data suggest that the chlamydial inhibitor in amniotic fluid does not pass through 0.45 microns pores and is larger than the bacterial inhibitor that was reported to be a peptide of low molecular weight.

Amniotic Fluid↗

Studies on human sexual development. III. Fetal pituitary and serum, and amniotic fluid concentrations of LH, CG, and FSH.

Fetal sera (9-20 weeks fetal age, n = 80) and pituitary glands (9.5-20 weeks, n = 36) obtained from hysterotomy specimens, and amniotic fluids (amniocentesis; 8-40 weeks, n = 123) were assayed for FSH, LH (betaLH assay) and CG (betaCG assay). Results are expressed as mass of pure hormone. Prior to 12 weeks fetal age, pituitary, serum and amniotic fluid concentrations of LH and FSH were low or unmeasurable. In contrast, levels of CG in serum and in amniotic fluid were clearly measurable prior to 12 weeks. There was a definite CG peak at 11-14 weeks with levels up to 550 ng/ml in serum and 7400 ng/ml in amniotic fluid. Although LH levels began to rise at 12 weeks, when CG levels started to decline, serum levels of LH from 14-20 weeks in males (2-13 ng/ml) were still lower than the majority of CG levels at this time (6-115 ng/ml). These observations suggest that CG is the primary stimulus to the fetal Leydig cell which results in testosterone secretion (peak 11-17 weeks) and masculine differentiation of the genital tract. Significantly lower concentrations of both FSH and LH were observed in pituitary, serum and amniotic fluid between 12-20 weeks fetal age in males compared to females. This may be a result of feedback inhibition by the higher concentrations of testosterone in males at this time. Amniotic fluid FSH and LH concentrations correlated with their respective serum and pituitary values (P less than 0.01) indicating that amniotic fluid may provide a convenient index of fetal serum concentrations.

Amniotic Fluid↗

Total antioxidant capacity and reactive oxygen species in amniotic fluid.

OBJECTIVE: To identify the presence and/or absence of reactive oxygen species and total antioxidant capacity in amniotic fluid and to determine changes in the levels of reactive oxygen species and total antioxidant capacity with advancing gestational age of pregnancy and fetal or neonatal weights. METHODS: Amniotic fluid was collected from a total of 26 nonsmoking patients. Nine specimens were collected in the third trimester of pregnancy, and 17 specimens were collected in the second trimester. Amniotic fluid reactive oxygen species and total antioxidant capacity levels were determined by chemiluminescence and spectrophotometric assays, respectively. Reactive oxygen species and total antioxidant capacity levels were established and then compared for advancing gestational ages and estimated fetal weights or neonatal weights. RESULTS: Reactive oxygen species levels were present in some but not all specimens, and total antioxidant capacity was present in all specimens. Total antioxidant capacity but not reactive oxygen species levels increased from the second to the third trimester (347.0 mmol/L versus 776.0 mmol/L, P <.001). There was a positive Spearman correlation between total antioxidant capacity and gestational age (r = 0.72, 95% confidence interval 0.43, 1.0, P <.001) and between total antioxidant capacity and estimated fetal weights or neonatal birth weights (r = 0.70, 95% confidence interval 0.40, 1.0, P <.001). There was no correlation between reactive oxygen species and advancing gestational age or weight. CONCLUSION: Total antioxidant capacity levels are present in amniotic fluid at least as early as the second trimester and increase with advancing gestational age and fetal or neonatal weights. Reactive oxygen species levels are not necessarily present in amniotic fluid, and they do not appear to be influenced by gestational age or estimated fetal or neonatal weights.

Adult↗

Is there a relationship to dye determined or ultrasound estimated amniotic fluid volume adjusted percentiles and fetal weight adjusted percentiles?

OBJECTIVE: This study was undertaken to determine whether there is a correlation between neonatal birth weight and an estimated or dye-determined amniotic fluid volume. STUDY DESIGN: A prospective observational study of women undergoing an amniocentesis and subsequent delivery within 72 hours. Amniotic fluid volume was estimated using the amniotic fluid index (AFI) and single deepest pocket technique and calculated by the dye-dilution technique. RESULTS: One hundred seventy-five women participated in the study. Linear regression modelling showed no significant relationship between birth weight and amniotic fluid volume adjusted for gestational age (P=.062). Similarly, there was no evidence of an association between birth weight and a dye determined amniotic fluid volume (P=.180), fixed cutoffs for the AFI (P=.224), percentiles for the AFI (P=.112), or fixed cutoffs for the single deepest pockets (P=.867). CONCLUSION: Neonatal birth weight is not correlated with a dye-determined or ultrasound estimated amniotic fluid volume.

Adolescent↗

Influence of tobacco smoking on metallothionein isoforms contents in human placenta, amniotic fluid and milk.

SDS-PAGE was used for separation of heat-stable proteins in placenta, amniotic fluid and milk of women who smoke actively and those exposed to tobacco smoke (passive smokers). In the number of protein bands stained with Coomassie brilliant blue R-250, several bands were stained with silver nitrate. The presence of low-molecular band with molecular weight of 6.5 kDa, corresponding to the mobility of electrophoretic MT-1 was detected in placenta, amniotic fluid and milk. The involvement of MT-2 isoform was much lower, although more evident in amniotic fluid and milk of active smokers. In this group of women an enhanced concentration of the band with molecular weight of 25-30 kDa was observed in placenta, colostrum and milk on the second day after delivery; several protein bands emerged in the area of the same mobility in amniotic fluid; the presence of the band with molecular weight of 12.5 kDa and the absence of protein bands of 25-30 kDa were found in milk excreted on the third day post partum. Our results show the apparent differences between proteingrams of placenta, amniotic fluid and milk of active and passive smokers.

Adult↗

[Study of the amniotic fluid of sheep in the normal course of pregnancy and in abortion].

Cytologic, biochemical and immunoelectrophoretic studies were carried out of amniotic fluids in 100 ewes with normal pregnancy and 40 ewes that had miscarried. Each month of pregnancy a total of 20 and 8 animals of the two groups, respectively were studied. It was found that the biochemical and metabolic processes taking place in the fetus lead to changes in the amniotic fluids altering the pH value, the alkali reserve, the content of potassium, calcium, phosphorus, alkaline phosphatase as well as their bactericidal activity. More characteristic changes linked with pregnancy were observed in the cell composition of the amniotic fluids. With advancing in age the increase in cell count was accompanied (staining with Nile blue sulfate) with a rise of the "orange cell" content. The amniotic fluids of ewes with normal pregnancy were found to contain proteins which precipitated with hyper immune sera against blood serum and kidney, heart, and placenta proteins. In ewes that had miscarried the pH values of the amniotic fluids dropped in the months when abortions took place: 7.36, 7.11, 6.90, 6.80 and 6.90, as against 7.41, 7.36, 7.28, 7.17 and 7.18, respectively. Along with pH the alkali reserve also dropped to 37.9 in the first month and 14.20 in the fifth month. In ewes that had miscarried in the 2nd, 3rd, and 4th month these values were 18.90, 14.90, and 13.80 cu. cm, respectively. In cases of abortions the protein composition of the amniotic fluids showed higher levels of the alfa and beta globulins.

Abortion, Veterinary↗

Longitudinal study of the amniotic fluid index in post-dates pregnancy.

Amniotic fluid (AF) was measured in 511 post-dates pregnancies (at least 41 weeks of gestational age) with the use of the AF index. Ultrasonographic evaluations were conducted on a semiweekly basis. Only patients with reliable gestational ages calculated from certain last menstrual period and confirmed by early sonographic estimates participated in the study. Oligohydramnios (AF index of 5.0 cm or less) was detected in 11.5% of the study population. Longitudinal data were available from 121 patients who demonstrated a mean 25% decrease in AF index per week beyond 41 weeks' gestation. The longitudinal change in AF index was statistically significant (P less than .0005). Amniotic fluid index measurements ranged from 1.7-24.6 cm, with a mean of 12.4 at 41 weeks' gestation. Compared with previous cross-sectional studies, this longitudinal study provides a more accurate estimate of changes in AF levels as a function of gestational age.

Amniotic Fluid↗

Clinical implications of amniotic fluid alpha-fetoprotein in twin pregnancy.

Amniotic fluid alpha-fetoprotein (AF-AFP) was measured in 68 pairs of normal twins. Acetylcholinesterase (ACHE) was measured in 23 of those pairs. A significant difference in AF-AFP levels was found in 9/49 (19.4%) of twin pairs of the same sex and 9/19 (47.3%) of twin pairs of opposite sexes (P = .015). Differences in fetal size or gestational age at amniocentesis did not influence the rate of discordant AFP results. ACHE activity was identical in all amniotic fluid samples in twin pairs. In five additional patients, fetal anomalies (three), fetal death or fetal blood in the sample could have affected AFP levels, which were found to be discordant in three of them. Using fetal sex to represent zygosity, the study indicated that discordance in AF-AFP is more common in dizygous twins. The odds ratio for having a discordant AFP result if twins are of different sexes was 4.0. Implications of our data are: (1) in same-sex twins, discordance between sacs is less common, even if only one fetus is affected; (2) the difference in AFP results in twins of the same sex and of different sexes suggests variability in AFP transfer between twin gestational sacs; and (3) ACHE readily diffuses between sacs and cannot be used to determine which twin has an abnormality.

Acetylcholinesterase↗

Establishment of reference values of five amniotic fluid enzymes. Analytical performances of the Hitachi 911. Application to complicated pregnancies.

OBJECTIVES: to evaluate the analytical performances of the Hitachi 911 analyzer (Roche Diagnostics, F) for gamma-glutamyl-transferase, 5'nucleotidase, amylase, aspartate-amino-transferase and total-alkaline-phosphatase assay in amniotic fluid. To establish reference intervals for these five enzymes throughout gestation. To determine their antenatal diagnostic value. DESIGN AND METHODS: amniotic fluid samples were collected between 14 and 35 weeks of gestation. Weekly numbers ranged from 31 to 92. Techniques developed for serum enzyme assays were applied to amniotic fluid. Two pools of amniotic fluid containing low and high marker levels were used. Within-day and between-day variations were calculated, together with the limits of detection and linearity. Reference ranges were established on 508 amniotic fluid samples, including 23 samples from pregnancies with chromosomal aberrations, 14 with gastrointestinal tract defects and 5 with gastroschisis. RESULTS: the assay technique for total-alkaline-phosphatase assay had to be adapted to amniotic fluid, but no adaptation was necessary for the other markers. The within-day CV ranged between 2.2 and 11.2% for low-level samples and from 1.1 to 3.4% for high-level samples. The between-day CV ranged from 6.3 to 13.3% for low-level samples and 1.2 to 4.7% for high-level samples. Total-alkaline-phosphatase activity fluctuated throughout gestation. Amylase levels and aspartate-amino-transferase levels increased whereas gamma-glutamyl-transferase and 5'nucleotidase levels fell until delivery. All trisomy 18 and trisomy 13 pregnancies, 65% of Down's syndrome pregnancies and all pregnancies with digestive tract defects were associated with marked changes in the level of at least one enzyme. CONCLUSION: The Hitachi 911 is suited to rapid, reliable quantification of amniotic fluid enzymes with only minor adaptation. Useful reference intervals can be obtained throughout gestation. Gamma-glutamyl-transferase, 5'nucleotidase, total-alkaline-phosphatase and amylase assay can help to confirm echographic evidence of bowel disorders and thereby improve patient management especially in case of gastroschisis.

Alkaline Phosphatase↗

Inhibition of mitogen and allogeneic stimulated lymphocyte growth by human amniotic fluid: lack of correlation between alpha-fetoprotein level and in vitro immunosuppression.

The influence of human amniotic fluid from women at 16-18 wk gestation on the in vitro proliferation of human peripheral lymphocytes stimulated with various mitogens (PHA, Con A and PWM) and with allogeneic cells in the one- and two-way mixed lymphocyte reaction (MLR) was studied. It was seen that amniotic fluid inhibits nonspecifically the responsiveness of the various subpopulations of cells induced to proliferate. It was also demonstrated that the suppression depends on the concentration of the immunosuppressive agent(s), since diluted amniotic fluids showed less inhibitory capacity. In order to detect a possible immune action for alpha-fetoprotein (AFP), the levels of the protein in the amniotic fluid supplemented cultures were correlated with the inhibitory properties of the fluids. No correlation was encountered, indicating that AFP is not mainly responsible for the in vitro immunosuppressive properties of human amniotic fluid on the in vitro test of cell-mediated immunity here utilized.

Amniotic Fluid↗

Trisomy 18 and maternal serum and amniotic fluid alpha-fetoprotein.

Maternal serum and amniotic fluid alpha-fetoprotein levels were studied retrospectively in a total of 58 pregnancies with trisomy 18. In those pregnancies uncomplicated by either fetal exomphalos or neural tube defect the midtrimester maternal serum alpha-fetoprotein (MSAFP) levels were markedly reduced, the median value for 38 such pregnancies being 0.6 multiples of the median (MoM). Trisomy 18 with exomphalos was associated with a higher median MSAFP, but still within the normal range: 1.1 MoM, (nine pregnancies); trisomy 18 with exomphalos and neural tube defect (NTD) was associated with grossly raised levels: median MSAFP was 4.5 MoM (three pregnancies). Amniotic fluid alpha-fetoprotein (AFAFP) levels were normal in uncomplicated trisomy 18 pregnancies: median AFAFP, for 19 pregnancies, was 1.1 MoM. Exomphalos alone, or together with neural tube defect, was associated with greatly elevated levels of AFAFP; for exomphalos alone median AFAFP was 9.59 MoM (four pregnancies), and for exomphalos with neural tube defect the median AFAFP was 23.95 Mom (three pregnancies). Screening with low and high MSAFP, routine ultrasound, and amniocentesis on all women aged 35 years or over, together might identify over 50 per cent of pregnancies with trisomy 18.

Adult↗

Amniotic fluid tumor necrosis factor-alpha and interleukin-1 in a rabbit model of bacterially induced preterm pregnancy loss.

OBJECTIVE: The objective of this study was to determine whether the cytokines tumor necrosis factor-alpha, interleukin-1 alpha, and interleukin-1 beta were produced in the amniotic fluid of the rabbit after intracervical inoculation with Escherichia coli. STUDY DESIGN: Timed pregnant rabbits on day 21 (70% of gestation) were inoculated with a hysteroscope intracervically with 10(4) to 10(5) colony-forming units Escherichia coli or sterile saline solution. Escherichia coli-inoculated animals (N = 16) were put to death at 4, 8, 12, and 16 hours after inoculation. Control animals (N = 6) were put to death at similar intervals. At death, cultures were taken from endometrium, amniotic fluid, peritoneum, and blood. Amniotic fluid was collected and assayed for tumor necrosis factor bioactivity by a modified fibroblast cytotoxic assay in L929 cells, for interleukin-1 alpha, and interleukin-1 beta with a specific radioimmunoassay, and for prostaglandin E2 and prostaglandin F2 alpha by radioimmunoassay. RESULTS: Levels of amniotic fluid tumor necrosis factor-alpha, interleukin-1 alpha, and interleukin-1 beta were elevated as early as 4 hours after inoculation in some animals and by 12 to 16 hours after inoculation in all. Levels of all three cytokines correlated significantly with time from intracervical inoculation with Escherichia coli (p < 0.05). Levels of amniotic fluid prostaglandin E2 and prostaglandin F2 alpha correlated significantly with time from intracervical inoculation with Escherichia coli (p < 0.05). CONCLUSIONS: Levels of tumor necrosis factor-alpha, interleukin-1 alpha, interleukin-1 beta, prostaglandin E2 and prostaglandin F2 alpha are elevated in the amniotic fluid of rabbits after intracervical inoculation with Escherichia coli. Similarity exists between elevations of amniotic fluid cytokines in this model and in cases of intraamniotic infection and preterm labor unresponsive to tocolytics in humans. Modulation of cytokines may offer a strategy for improvement of outcome in this experimental model of infection-induced pregnancy loss.

Abortion, Veterinary↗

Circulating intercellular adhesion molecule-1 in amniotic fluid, maternal serum alpha-fetoprotein levels, and intrauterine growth retardation.

OBJECTIVE: Our purpose was to determine if circulating intercellular adhesion molecule-1, a marker of chronic inflammation, is present in amniotic fluid in midtrimester, is increased in patients with elevated maternal serum alpha-fetoprotein level, and is associated with intrauterine growth retardation. STUDY DESIGN: Amniotic fluid circulating intercellular adhesion molecule-1 levels were assayed by enzyme-linked immunoassay in 273 samples obtained by midtrimester amniocentesis in gestations involving a single, structurally normal fetus. The control group consisted of 108 patients with normal maternal serum alpha-fetoprotein levels and 165 patients with elevated levels. Intrauterine growth retardation was diagnosed if birth weight was < 10th percentile for the clinically estimated gestational age. RESULTS: Circulating intercellular adhesion molecule-1 was detectable in amniotic fluid in 105 of 273 samples (38%). In the control group it was detectable in amniotic fluid in seven of 108 (6%). In patients with elevated maternal serum alpha-fetoprotein 97 of 164 (59%) had detectable levels (p < 0.001). Of the 273 cases 38 (14%) had intrauterine growth retardation. Of these 23 (59%) had detectable circulating intercellular adhesion molecule-1 levels (p < 0.001). Of the seven cases of intrauterine growth retardation with normal maternal serum alpha-fetoprotein levels, one (14%) had detectable circulating intercellular adhesion molecule-1. Of the 31 cases of intrauterine growth retardation with elevated maternal serum alpha-fetoprotein 22 (71%) had detectable circulating intercellular adhesion molecule-1. When circulating intercellular adhesion molecule-1 was detectable in amniotic fluid, increasing levels was significantly related to decreasing gestational age at delivery (p < 0.005). CONCLUSIONS: Midtrimester amniotic fluid from normal pregnancies does not generally contain detectable circulating intercellular adhesion molecule-1. Detectable amniotic fluid levels are significantly related to a birth weight < 10th percentile at delivery and to elevated midtrimester maternal serum alpha-fetoprotein levels. Increasing circulating intercellular adhesion molecule-1 levels are related to shortened length of gestation. This test may contribute to risk assessment for intrauterine growth retardation and prematurity. Circulating intercellular adhesion molecule-1 is a known marker of inflammatory processes; its further study may also improve understanding of the pathophysiologic mechanisms of certain cases of intrauterine growth retardation and prematurity.

Amniotic Fluid↗

Antibacterial activity of amniotic fluid against Staphylococcus aureus, Candida albicans and Brucella abortus.

Sixty amniotic fluid samples from sixty patients between 14--42 weeks gestation were studied for antibacterial activity against Staphylococcus aureus, Brucella abortus and antifungal activity against Candida albicans. Both antibacterial and antifungal activity of the amniotic fluid were observed. When antimicrobial activity was correlated with gestational age, it was found to increase with the period of gestation. Maximum antimicrobial activity of the amniotic fluid was found to be present in the samples obtained between 36--42 weeks of gestation. All samples before 20 weeks gestation showed week antibacterial activity against Staphylococcus aureus, but comparatively strong reaction against Candida albicans and Brucella abortus.

Amniocentesis↗