Early amniocentesis: alphafetoprotein levels in amniotic fluid, extraembryonic coelomic fluid and maternal serum between 8 and 13 weeks.

OBJECTIVE--The aim was to establish a normal range of alphafetoprotein (AFP) concentrations in amniotic fluid from 8 to 12 weeks gestation, and to determine any difference between AFP levels in amniotic fluid and extraembryonic coelomic fluid. DESIGN AND SUBJECTS--150 women had a transvaginal ultrasound guided amniocentesis before termination of an apparently normal first trimester pregnancy. Separately identified samples of amniotic fluid and extraembryonic coelomic fluid were obtained and assayed by radioimmunoassay for AFP. RESULTS--In amniotic fluid, very high levels of AFP were present at 8 weeks, levels falling rapidly up to 10 weeks after which there was a slight rise. Thus over the period 8 to 10 weeks, there was a significant inverse correlation between amniotic fluid AFP and gestational age (r = 0.67; P less than 0.001). In extraembryonic coelomic fluid, by contrast there was no trend in AFP relative to gestational age. CONCLUSIONS--The rapidly changing levels of AFP from 8 to 10 weeks as well as the small volume of the amniotic cavity makes the use of amniocentesis impracticable before 11 weeks gestation. The lack of any relation between AFP levels in amniotic fluid and extraembryonic coelomic fluid emphasises the importance of identifying the site of amniocentesis in the first trimester.     

First-trimester amniotic fluid and extraembryonic coelomic fluid acetylcholinesterase electrophoresis.

Acetylcholinesterase (AChE) gel electrophoresis was performed on samples of amniotic fluid and extraembryonic coelomic fluid obtained by high resolution transvaginal ultrasound-guided amniocentesis from 38 women between 8 and 12 weeks of pregnancy. AChE was positive in 33 per cent (12/36) of the amniotic fluid samples; the percentage of positive results decreased as gestation advanced. AChE was positive in 32 per cent (9/28) of the extraembryonic coelomic fluid samples. In 81 per cent (21/26) of matched samples, the AChE results were identical in the two fluids. Amniotic fluid and extraembryonic coelomic fluid AChE electrophoresis cannot be used to diagnose neural tube defects prior to 12 weeks of gestation.     

Erythropoietin levels in amniotic fluid and extraembryonic coelomic fluid in the first trimester of pregnancy.

OBJECTIVE: The aim was to measure erythropoietin levels in amniotic fluid and extraembryonic coelomic fluid from 7-12 weeks' gestation. SUBJECTS: Twenty healthy women with ultrasonographically normal first trimester pregnancies prior to surgical termination. METHODS: Paired samples of amniotic fluid and extraembryonic coelomic fluid were collected by transvaginal ultrasound guided needling. Erythropoietin was measured in both pregnancy fluids using a radioimmunoassay. RESULTS: There was a highly significant difference between erythropoietin levels in extraembryonic coelomic fluid (median level 15.45 mU/ml; range 6.8-32.1 mU/ml) and those in amniotic fluid (median 5.0 mU/ml; range < 5.0-5.8 mU/ml) (P < 0.0001; Mann-Whitney U-test). The levels of erythropoietin in maternal serum (median 15.4 mU/ml; range 5.6-29.4 mU/ml) were similar to those in the extra-embryonic coelom (P = 0.81; Mann-Whitney U-test). No relation was demonstrated between erythropoietin levels in amniotic fluid or coelomic fluid and stage of gestation. CONCLUSION: High levels of erythropoietin in coelomic fluid suggests that the hormone is involved in the process of human extraembryonic erythropoiesis. The exact regulatory role remains unknown.     

The coelomic cavity — A reservoir for metals

OBJECTIVE: The concentrations of metals in fluids surrounding the first trimester fetus were measured.STUDY DESIGN: Atomic absorption spectrometry was used to measure concentrations of metals in matched samples of amniotic and extraembryonic coelomic fluids in 17 women between 9 and 12 weeks of pregnancy.RESULTS: Concentrations of calcium, magnesium, iron, copper, and manganese (but not zinc, cadmium, or lead) were significantly higher in coelomic than in amniotic fluid. There was no significant difference between levels of iron, manganese, and lead to controls and amniotic fluid or between concentrations of manganese, cadmium, and lead in controls and coelomic fluid. There was no relationship between the concentrations of each metal in amniotic and coelomic fluid.CONCLUSION: The extraembryonic coelom is an important site of concentration of metals in early pregnancy. This might represent a store of metals essential for normal embryonic and fetal development or constitute a defense mechanism against environmental conditions adverse to the fetus.     

Microvillar enzyme activity in amniotic fluid, extraembryonic coelomic fluid and maternal serum in the first trimester of pregnancy.

The activities of two microvillar enzymes, gamma-glutamyl transpeptidase and total alkaline phosphatase, have been measured in samples of amniotic fluid and extraembryonic coelomic fluid obtained by high-resolution transvaginal ultrasound-guided amniocentesis from 40 women between 7 and 12 weeks of gestation. There was a highly significant difference between gamma-glutamyl transpeptidase activity in amniotic fluid (median level 31 U/l; range 2-409 U/l) and extraembryonic coelomic fluid (median level 2 U/l; range less than 2-16 U/l) (P less than 0.001; Mann-Whitney U-test). Alkaline phosphatase activity was not detected in 84% of amniotic fluid samples and 97% of extraembryonic coelomic fluid samples. No difference was found between total alkaline phosphatase activity in these fluids (P = 0.14; Mann-Whitney U-test). Enzyme activities in amniotic fluid increased with gestational age. A significant linear correlation was found between amniotic fluid gamma-glutamyl transpeptidase activity and stage of gestation (r = 0.86; P less than 0.001) and total alkaline phosphatase activity in amniotic fluid and stage of gestation (r = 0.66; P less than 0.001).     

Human chorionic gonadotrophin and alpha-fetoprotein levels in matched samples of amniotic fluid, extraembryonic coelomic fluid, and maternal serum in the first trimester of pregnancy.

Separately identified samples of amniotic fluid and extraembryonic coelomic fluid obtained by high resolution transvaginal ultrasound-guided amniocentesis from 32 women between 7 and 12 weeks of pregnancy were analysed for human chorionic gonadotrophin (hCG) and alpha-fetoprotein (AFP). There was a highly significant difference between the hCG levels in amniotic fluid (median level 6.3 U/ml; range 1.6-310.0 U/ml) and those in extraembryonic coelomic fluid (median level 400.0 U/ml; range 135.0-2250.0 U/ml) (p less than 0.001; Mann-Whitney U-test). The levels of AFP were very similar in amniotic fluid (median 26.0 kU/ml; range 10.0-116.5 kU/ml) and extraembryonic coelomic fluid (median level 24.1 kU/ml; range 12.4-94.4 kU/ml).     

Maternal and fetal levels of methionine and homocysteine in early human pregnancy

Objective To investigate methionine metabolism during normal human embryonic development by measuring levels of methionine and total homocysteine in samples of maternal serum, extra-embryonic coelomic fluid, and amniotic fluid.
Design Cross-sectional observational study.
Setting Collaboration between St Bartholomew's Hospital, London, and the University Hospital of Nijmegen in The Netherlands.
Participants Twenty-three women with uncomplicated pregnancies between 8 and 12 weeks of gestation before surgical termination of an ultrasonographically normal fetus.
Methods Maternal serum samples were collected prior to surgery. Samples of extra-embryonic fluid and amniotic fluids were obtained by transvaginal ultrasound-guided coelocentesis and amniocentesis. Methionine was measured using an aminoacid analyser and total homocysteine by high performance liquid chromatography.
Results Levels of methionine were four times higher in extra-embryonic coelomic fluid and twice as high in amniotic fluid compared with maternal serum. In contrast, the total homocysteine concentrations were much lower in both extra-embryonic coelomic fluid and amniotic fluid than in maternal serum. All differences were significant (   P 0.01  ).
Conclusions The comparatively high concentrations of methionine in extra-embryonic coelomic fluid and amniotic fluid, and the concomitant low levels of total homocysteine in these fluids, suggest a role for methionine metabolism during early human pregnancy.     

Folate and vitamin B12 concentrations in maternal and fetal blood, and amniotic fluid in second trimester pregnancies complicated by neural tube defects.

OBJECTIVE: To investigate maternal and fetal folate and vitamin B12 concentrations in pregnancies affected by neural tube defects (NTD). DESIGN: Measurement of folate and vitamin B12 concentrations in amniotic fluid, fetal blood and maternal blood samples in midgestation. SUBJECTS: 32 women undergoing termination of pregnancy at 14-21 weeks gestation for social reasons (n = 24) or for fetuses with neural tube defects (n = 8). INTERVENTIONS: Fetoscopy before intra-amniotic injection of prostaglandins. RESULTS: In normal pregnancies there was a positive correlation between maternal and fetal serum folate, and the fetal serum and red blood cell folate concentrations were higher than the maternal. There were no differences in amniotic fluid, maternal blood or fetal blood folate concentrations between pregnancies with NTD and normal pregnancies. Although amniotic fluid vitamin B12 was lower in pregnancies with NTD, maternal serum vitamin B12 concentration was not reduced. CONCLUSION: In this small group of pregnancies with NTD at mid-gestation there is no evidence to suggest folate or vitamin B12 deficiency.     

ADAM12-s in coelomic fluid and maternal serum in early pregnancy

OBJECTIVES: ADAM12-s is a placental protein. In early pregnancy, reduced maternal levels of ADAM12-s have been reported in association with foetal trisomy 21 or 18 and in cases that subsequently develop pre-eclampsia and foetal growth restriction. The aim of this study is to investigate the distribution of ADAM12-s in early pregnancy by comparing its concentration in maternal serum, amniotic fluid and coelomic fluid. METHODS: Coelomic fluid was obtained by coelocentesis from 13 singleton pregnancies with live foetuses at 6.9-9.3 weeks of gestation. Maternal serum was also obtained in all cases and in six cases amniotic fluid was also obtained. The concentration of ADAM12-s was measured by dissociation enhanced lanthanide fluoro-immunoassay. RESULTS: The median concentration of ADAM12-s in maternal serum was 132.7 (range 33.8-254.5) ng/mL and in coelomic fluid it was 10.5 (range 1.3-15.8) ng/mL; there were no detectable levels in five of the six amniotic fluid samples. The concentration of maternal serum ADAM12-s increased significantly with gestation (r = 0.862, p < 0.0001). There was no significant association between coelomic fluid ADAM12-s and either gestation (r = 0.255, p = 0.401) or maternal serum ADAM12-s (r = 0.302, p = 0.316). CONCLUSION: The distribution of ADAM12-s in maternal serum and the early embryonic fluid compartments is consistent with its syncytiotrophoblastic origin.     

Selenium status in pregnancy: studies in amniotic fluid from normal pregnant women

Selenium concentrations were determined in amniotic fluid samples obtained from 111 healthy normal pregnant women (median age, 27.5 years) between 12 and 42 weeks of gestation using hydride generation technique coupled with atomic absorption spectroscopy. There was a gradual decrease in the amniotic fluid selenium concentration with the progress of pregnancy. The negative correlation between the gestational age and amniotic fluid selenium concentration was highly significant (p less than 0.001). The implications of these findings in normal pregnancy are discussed.     

Coelomic fluid leptin concentration in normal first-trimester pregnancies and missed miscarriages

OBJECTIVE: Investigation of the possible role of leptin in early pregnancy failure. METHODS: Leptin concentration was measured in maternal serum, coelomic fluid and amniotic fluid from 15 singleton pregnancies with live fetuses and 7 missed miscarriages at 7-10 weeks of gestation. RESULTS: In the pregnancies with live fetuses, the median leptin concentration was significantly higher in coelomic fluid (median 33.1 ng/ml) than in maternal serum (median 8.1 ng/ml) or amniotic fluid (median 0.5 ng/ml). In the pregnancies with missed miscarriage, compared to those with live fetuses, the median leptin concentration in coelomic fluid was higher (median 45.3 ng/ml), but in maternal serum it was not significantly different (median 5.5 ng/ml). CONCLUSIONS: The high coelomic fluid leptin concentration suggests that embryonic death may be preceded by impaired oxygenation of the placenta that stimulates production of leptin.     

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.     

Physiological distribution of placental growth factor and soluble Flt-1 in early pregnancy

OBJECTIVE: To examine the distribution of placental growth factor (PlGF), vascular endothelial growth factor (VEGF) and soluble VEGF receptor-1 (sFlt-1) in maternal and embryonic fluid compartments in early pregnancy. METHOD: The concentrations of PlGF, VEGF and sFlt-1 were measured in coelomic fluid and maternal serum from 16 singleton pregnancies at 7.0-9.3 weeks. In six cases, amniotic fluid was also examined. RESULTS: The median concentration of PlGF was 14.1 (range 8.9-27.6) pg/mL in maternal serum, 13.9 (range 9.5-31.4) pg/mL in coelomic fluid and 8.9 (range 3.9-15.3) pg/mL in amniotic fluid. The concentration of PlGF increased between 7.0 and 9.3 weeks in maternal serum (p = 0.001) and decreased in coelomic and amniotic fluid (p = 0.001). The median concentration of sFlt-1 was 8561 (range 6724-10 673) pg/mL in coelomic fluid, 523 (range 244-986) pg/mL in maternal serum, 30 (range 12-83) pg/mL in amniotic fluid (p = 0.0001), and it did not change significantly with gestation. VEGF was undetectable in most of the samples, and therefore, no further analysis was performed. CONCLUSION: PlGF and sFlt-1 are present in the maternal and fetal fluid compartments in very early pregnancy, and their distribution is consistent with their site of production and the local conditions of transport.     

Oxytocinase activity in human amniotic fluid and its relationship to gestational age

Oxytocinase (EC 3.4.11.3) activity in amniotic fluid samples obtained from 200 normal pregnant women (mean age, 28 years) between 14 and 40 weeks of gestation and during active labor was assayed using S-benzyl-L-cysteine-p-nitroanilide (BCN) and L-leucine-p-nitroanilide (LN) separately as substrates. With both substrates, amniotic fluid oxytocinase activity correlated inversely with gestational age; the level of the enzyme, which was highest in early pregnancy, decreased exponentially to a minimum near term and during labor. However, whether oxytocinase activity in the amniotic fluid is of any significance for the maintenance and/or termination of pregnancy remains to be established.     

Reduced nitric oxide in amniotic fluid of patients with chorioamnionitis

OBJECTIVE: Levels of nitric oxide (NO) and cytokines were assessed in amniotic fluid obtained from patients with severe chorioamnionitis (CAM) and appropriate controls. METHODS: Amniotic fluid was obtained from 12 patients with CAM (17-24 weeks of gestation) and 89 patients undergoing diagnostic amniocentesis (16-18 weeks of gestation). The concentrations of NO, interleukin-6 (IL-6), and leukocyte elastase (LE) in amniotic fluid were then measured and compared. RESULTS: The concentrations of NO, IL-6, and LE were all higher in CAM cases than in normal pregnant women. Furthermore, an inverse correlation between NO and LE was suggested in the CAM group. CONCLUSIONS: These results indicate that in severe CAM, the action of NO might be reduced, not only due to blockage of action but also by degradation, despite increased production.     

Meconium-stained amniotic fluid across gestation and neonatal acid-base status

OBJECTIVE: To estimate whether the acid-base status of neonates born to women with meconium-stained amniotic fluid varies across gestation. METHODS: We carried out a retrospective cohort study of all pregnancies that were complicated by meconium-stained amniotic fluid in 2004. Cases were identified from a perinatal pathology database that contained data on all pregnancies complicated by meconium-stained amniotic fluid. Data abstracted from the charts included gestational age at delivery, umbilical arterial pH, birth weight, and the presence or absence of labor. Cases were stratified according to gestational age at delivery. The distribution of meconium-stained amniotic fluid across gestation was computed. The mean umbilical arterial pH values (with 95% confidence intervals) across gestation were assessed by analysis of variance. RESULTS: The mean umbilical arterial pH in women with meconium-stained amniotic fluid did not differ across gestation. The overall incidence of meconium-stained amniotic fluid was 12.0% (766 of 6,403 deliveries). The rates of meconium-stained amniotic fluid increased from 1.2% at 32 weeks to 100% at 42 weeks. CONCLUSION: The rising incidence of meconium-stained amniotic fluid with gestational age is consistent with the hypothesis that fetal maturation is a major etiologic factor in meconium passage. Also, the lack of variation of mean umbilical arterial pH across gestation suggests that fetal acidemia is not increased when meconium passage occurs earlier in pregnancy rather than at later gestational ages.     

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.     

Evaluation of neutrophile elastase and isoprostane 8epiPGF2alpha concentrations in maternal and umbilical cord blood serum and in amniotic fluid in pregnancies complicated by premature rupture of membranes

OBJECTIVES: To evaluate the total isoprostane 8-epi-PGF2alpha and neutrophil elastase (NE) concentrations in pregnancies complicated by premature rupture of membranes (PROM). MATERIAL AND METHODS: 128 pregnant women were divided into four groups: pregnancies complicated by PROM between 24.-36.(PPBP-N) and between 38 a 41 weeks of gestation (PPBP-D), uncomplicated pregnancies between 24-36 gestation weeks (K1) and pregnancies delivered by cesarean section (before uterine contractions had started) after 38 weeks (K2). The concentrations of NE and isoprostane 8-epi-PGF2alpha were measured in maternal serum, cord blood serum and in the amniotic fluid. RESULTS: The following study revealed higher concentrations of NE in maternal serum and in the amniotic fluid than in the umbilical cord blood in PROM cases, and lower amniotic fluid than maternal serum concentrations in the control groups. Also, the levels of isoprostane differentiated between compartments in particular groups. In both groups complicated with PROM, higher maternal serum and amniotic fluid NE concentrations than in controls were found. There were no differences in isoprostane levels between the groups. CONCLUSIONS: 1. Higher concentrations of NE in maternal blood serum and in the amniotic fluid than in the umbilical cord blood in PROM cases, as well as lower amniotic fluid than maternal serum concentrations in the controls, may be connected with pathogenesis of PROM. 2. Differentiated maternal serum, cord serum and amniotic fluid isoprostane concentrations may suggest various intensity of oxidative stress in particular compartments. 3. Lack of differences in maternal serum, cord serum and amniotic fluid isoprostane concentrations may suggest similar intensity of oxidative stress in cases with PROM and intact membranes.     

Serum and amniotic fluid lactic dehydrogenase in pregnant women

No significant differences were found when the serum and amniotic fluid total lactic dehydrogenase (LDH) and LDH isoenzymes in women with newborn infants involved by Rh hemolytic disease were compared with women with uninvolved infants. During the twenty-fourth to thirty-second weeks of gestation, the serum LDH 1 level was higher and the serum LDH 3 level was lower than levels of the nonpregnant control subjects. Serum LDH 3 levels were lower than those of the nonpregnant control subjects during the thirty-third to forty-second weeks of pregnancy. It was noted that there was an increase in amniotic fluid LDH 5 with increasing gestational age.