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PubMed1996

A novel calcium-binding protein in amniotic fluid, CAAF1: its molecular cloning and tissue distribution.

We found by using a 45Ca2+ overlay technique a large amount of Ca(2+)-binding activity in bovine amniotic fluid from which a novel calcium-binding protein (CaBP) was purified and is referred to as CAAF1 (calcium-binding protein in amniotic fluid-1), with an apparent molecular mass of 8 kDa determined by N-tris(hydroxymethyl)-methylglycine/ SDS-PAGE. It was structurally homologous with MRP/calgranulin proteins (MRP8/calgranulin A and MRP14/calgranulin B), members of the S100 protein family, which are abundantly found in the cytoplasm of granulocytes and macrophages. CAAF1 lacked the predicted signal peptide sequence, which is consistent with other CaBPs. The tissue and cellular distribution of CAAF1 was determined by monoclonal antibodies developed against this protein. Its immunoreactivity was found in squamous epithelial cells, neutrophils, and some macrophages throughout the fetal body. An especially characteristic staining pattern was obtained in the squamous epithelium, including that of the esophagus, skin and amnion: CAAF1 was detected in the suprabasal squamous epithelial cells undergoing differentiation, but not in the cells in the proliferating basal layer. Northern blot analysis also showed that CAAF1 mRNA was highly expressed in bovine fetal esophagus and skin. On the other hand, our ELISA studies showed that CAAF1 protein was present in amniotic fluid at a concentration of about 120 nM, which was over 30 times as high as that in the fetal serum. These results suggested that CAAF1 is one of the stage-specific proteins in the differentiation of squamous epithelial cells, and that CAAF1 is preferentially produced by fetal squamous epithelial cells, including epidermal keratinocytes and amniotic epithelial cells, and it is stored in the amniotic fluid during embryogenesis.

Amino Acid Sequence↗
PubMed1975

[Amniotic fluid arginase in women at term].

Study on 34 cases. A considerable arginasic activity is observed in amniotic fluid in women at the end of pregnancy. This activity is weak, but increased by Mn2+ (about 5 to 8 times). The total amniotic fluid is more active than the surpernatant, the deposit of which has been eliminated. An important part of arginasic activity results from the elements of the deposit (amniotic cells, foetal cells and eventualy erythrocytes in hemorragic amniotic fluid).

Amniotic Fluid↗
PubMed1994

Concentrations of vitamins A and E in amniotic fluid, extraembryonic coelomic fluid, and maternal serum in the first trimester of pregnancy.

Paired samples of amniotic fluid and coelomic fluid were obtained by transvaginal ultrasound guided amniocentesis from 15 women with an ultrasonographically normal pregnancy between 8 and 12 weeks' gestation. Vitamins A and E were measured in the two pregnancy fluids and in maternal serum by high performance liquid chromatography with detection by ultraviolet absorption and fluorimetry respectively. Concentrations of vitamins A and E were higher in maternal serum than in coelomic fluid and were generally undetectable in amniotic fluid. All differences in concentration were significant. The vitamin E/cholesterol ratio was similar in maternal serum and coelomic fluid. No correlation was shown between the vitamin concentrations in the three fluids. These findings suggest that the coelomic cavity plays a part in the maternofetal exchange of these vitamins.

Amniotic Fluid↗
PubMed1986

Determination of cholinesterase and acetylcholinesterase in amniotic fluid. Uses in prenatal diagnosis and quality control.

The determination of acetylcholinesterase (AChE) has been shown to be as specific as alphafetoprotein (AFP) for the prenatal detection of open neural tube defects although AFP remains the method of choice. This paper describes a semi-automated technique for the analysis of acetylcholinesterase in amniotic fluid that: A) reduces the cost of the procedure; B) allows for a larger number of samples to be run at a time; and C) provides for more accurate and reproducible procedures and results. Six fetuses with neural tube defects (2 with gastroschisis and 3 where one twin was dead) were detected and found to have elevated AChE, TChE and 2 bands by electrophoresis. Quality control procedures using both pure enzyme and amniotic fluid with low and high levels of the enzyme are described. The analysis of 340 amniotic fluids of normal pregnancies indicates that the normal value for AChE is 5.17 +/- 2.63 mU/ml (97% confidence interval for the mean 4.84-5.49 mU/ml. A group of 27 abnormal pregnancies provides evidence that fetal vomiting and regurgitation, fetal demise, multiple cysts syndrome, idiopathic IUGR, arthrogryposis multiplex, hydrocephaly (stenosis of aqueductus), trisomy 21, trisomy 18, hydronephrosis, pyloric stenosis, heart malformation, ectopia cordis and multiple gestation produce elevated levels of pseudocholinesterase (PChE) in amniotic fluid. The use of pseudocholinesterase levels in amniotic fluid for prenatal diagnosis is proposed and discussed in view of its elevated levels in abnormal pregnancies where AChE is normal. The normal values for PChE are 23.86 mU/ml (mean) and 5.83 for standard deviation. Electrophoretic analysis was performed on all samples with values higher than one standard deviation above the mean.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholinesterase↗
PubMed1982

Determination of phosphatidylglycerol in amniotic fluid by a simple one-dimensional thin-layer chromatography method.

Phosphatidylglycerol (PG) in amniotic fluid is the second important component of lung surfactant phospholipids and may be clinically useful in assessing fetal lung maturity in utero. Although methods for PG determination are available, there are shortcomings in clinical application. We developed an alternative reliable one-dimensional thin-layer chromatography(TLC) method for separating and quantitating PG in amniotic fluid. A mini-TLC plate (8 X 10 cm) was prepared from silica gel H containing 5% ammonium sulfate. The plate was first developed in tetrahydrofuron/dimethoxymethane/methanol/2N ammonium hydroxide (30.0:20.6:5.6:3.0 v/v) and then in chloroform/methanol (60.9, v/v) in the same direction. PG was clearly separated from other phospholipids and neutral lipids, even when large amounts of other phospholipids were present on the TLC plate. The density of the charred PG was directly proportional to the amount of PG up to 8 nmol. The content of PG in nmol in the specimen can be quantitated by comparing with a standard PG. Up to 10% of blood serum or 3% meconium showed no detectable PG, nor did these substances affect PG quantitation in amniotic fluid. This method is sensitive and accurate. It is also time-saving and economical.

Amniotic Fluid↗
PubMed1980

Intra-amniotic or intravenous injection of dehydroepiandrosterone sulphate: simultaneous changes in steroid levels in amniotic fluid and maternal serum.

Levels of unconjugated oestradiol-17 beta (E2) and testosterone (T) and total oestriol (E3) were measured hourly to 6 h after intra-amniotic (i.a.) (100--200 mg) or intravenous (i.v.) (100 mg) injection of dehydroepiandrosterone sulphate (DHEA-S) in a series of twenty-six women with mid-trimester abortion. Amniotic fluid E2 and T levels were elevated significantly 1--6 h after the i.a. injection of DHEA-S, while the E3 level in amniotic fluid did not change. The i.a. administration also induced significant rises in serum E2 and T levels between 1 and 6 h and in the serum E3 level between 2--6 h. The i.v. injection of DHEA-S was accompanied by significantly elevated serum levels of E2 and T, while the E3 concentration remained unchanged. It appears that following maternal i.v. administration or an amniotic fluid injection, DHEA-S is rapidly converted to E2 and T, and these accumulate in both the maternal circulation and the amniotic fluid. E3, however, is formed only after i.a. administration of DHEA-S, and is selectively transported to the maternal blood.

Amnion↗
PubMed1987

Ultrasound placental grading and amniotic fluid quantitation in prolonged pregnancy.

Serial ultrasound examinations were performed every three to five days to monitor 200 singleton pregnancies with gestations of 42 weeks or longer. Scanning was used to measure the deepest amniotic fluid pool and to grade placental echogenic changes. Oligohydramnios, defined as an amniotic fluid pool less than 30 mm, was detected in 33 patients (17%) and was an indication for intervention. Grade III placenta was found in 53 patients (27%), and no grade could be assigned in 13 (7%). Intrauterine growth retardation and meconium-stained or absent amniotic fluid at amniotomy occurred significantly more often in association with oligohydramnios than with grade III placenta, which was found before spontaneous labor in only 24% of cases. Ultrasound placental grading appears to offer little useful additional information to amniotic fluid quantitation in satisfactory monitoring of prolonged pregnancies.

Amniotic Fluid↗
PubMed1977

[Antibacterial and antimycotic activity of the amniotic fluid against selected germs (author's transl)].

60 samples of amniotic fluid from 60 patients were obtained between 14 and 42 weeks gestation by dates and tested for their antibacterial activity against staphlococcus aureus strains and brucella abortus and their antimycotic activity against Candida albicans. The antibacterial and antimycotic activity of the amniotic fluid was confirmed. This activity increases steadily from the 15th week and reaches its maximum around term between 36 and 42 weeks. The amniotic samples obtained at 20 weeks gestation showed a weak antibacterial activity especially against staphlococcus aureus of human origin. The above results are important because of the increasing frequency of early and late diagnostic amniocentesis in pregnancy.

Amniotic Fluid↗
PubMed1986

Qualitative amniotic fluid volume versus amniocentesis in predicting infection in preterm premature rupture of the membranes.

Qualitative amniotic fluid volume assessment and amniocentesis were performed on admission in 54 patients who presented with premature rupture of the membranes and no clinical signs of infection or labor. Comparison of these two methods in predicting infection outcome--as reflected by the development of clinical amnionitis and/or neonatal sepsis--suggests them to have the same efficacy in predicting infection outcome in patients with premature rupture of the membranes. Qualitative amniotic fluid volume was found to have sensitivity 50%, specificity 92.8%, positive predictive value 66.6%, and negative predictive value 86.6%; amniocentesis was found to have sensitivity 58.3%, specificity 88%, positive predictive value 58.3%, and negative predictive value 88%. The use of ultrasonically estimated amniotic fluid volume could replace or be used in addition to amniocentesis in identifying patients with ruptured membranes at particular risk for developing infection.

Adult↗
PubMed1983

[Secretory immunoglobulin A (S-IgA) in amniotic fluid and in pharyngeal mucus and urine of newborn infants].

S-IgA was estimated in amniotic fluid and in mucus samples of pharyngeal cavities and urine samples of newborns for assistance of the hypothesis that there is a connection between S-IgA content in amniotic fluid and fetal pulmonary maturity. It was used an antiserum against human secretory component and an S-IgA standard in the single radial immunodiffusion according to Mancini and coworkers. High S-IgA concentrations in the mucus samples of pharyngeal cavities refer to fetal bronchial-pulmonary system as the probable essential part in the synthesis of S-IgA established in amniotic fluid of the 3rd trimenon in pregnancy.

Amniotic Fluid↗
PubMed1981

Alpha-fetoprotein and beta-human chorionic gonadotropin in amniotic fluid.

A series of data on amniotic fluid levels of alpha-fetoprotein (AFP) and beta-human chorionic gonadotropin (HCG) are presented. Both hormones were observed to decrease with advancing gestational age. The diagnostic importance of AFP levels is confirmed, and the very high levels of beta-HCG in twin gestations are documented. A statistically significant correlation between the concentration of both hormones is demonstrated.

Amniotic Fluid↗
PubMed1999

Vascular endothelial growth factor is bound in amniotic fluid and maternal serum.

To study vascular endothelial growth factor (VEGF) and placenta growth factor (PlGF) concentrations and their possible binders, serum from 22 non-pregnant and 55 pregnant women (15 at weeks 10-13; 40 at term), umbilical vein (n = 24) and artery (n = 13) and amniotic fluid (a pool of 50 at weeks 15-17; 11 at term) were assessed for VEGF and PlGF by an enzyme-linked immunosorbent assay. In amniotic fluid and maternal serum VEGF concentrations were <16 ng/ml and added VEGF was not recovered. VEGF was detected in serum from mothers post-partum (137 +/- 142 ng/l, mean +/- SD), umbilical artery (421 +/- 288 ng/l) and vein (502 +/- 339 ng/l) and non-pregnant controls (182 +/- 147 ng/l), and added VEGF was fully recovered. PlGF was detected in pregnancy serum (52 +/- 23 ng/l early pregnancy; 439 +/- 217 ng/l term pregnancy) and in amniotic fluid (early pregnancy 56 ng/l; term pregnancy 30 +/- 18 ng/l). PlGF was fully recovered in all samples. Gel filtration and isoelectric focusing revealed that in maternal serum and amniotic fluid [125I]VEGF was bound to a protein with an Mr of 400-700 kDa and an isoelectric point of approximately 8. This protein was not identical with alpha-2-macroglobulin (by an immunofluorometric assay), pregnancy zone protein or pregnancy associated plasma protein-A (by immunodiffusion). In conclusion, VEGF-binding activity is present in amniotic fluid and maternal blood. It disappears after delivery and is not detectable in fetal or non-pregnant serum.

Adolescent↗
PubMed1981

Apolipoproteins in human fetal blood and amniotic fluid in mid-trimester pregnancy.

To examine the potential for prenatal diagnosis of genetic lipoprotein metabolic defects (e.g. abetalipoproteinemia. Tangier disease) we determined the normal concentrations of apolipoproteins (apo) A-I, A-II, B, and E in mid-trimester amniotic fluid and fetal plasma. The concentrations of apo A-I and apo A-II in amniotic fluid were 1--2 per cent of the respective levels in the mother's plasma, whereas apo B and apo E were undetectable in amniotic fluid. In contrast to amniotic fluid, all four apolipoproteins were detectable in fetal plasma, and the levels of apo A-I, apo B and apo E were in the range observed in the mothers: 160.2 +/- 103.1, 59.8 +/- 35.7 and 5.7 +/- 3.5 mg/dl respectively (mean +/- SD, n = 13). The fetal plasma level of apo A-II (28.3 +/- 12.4 mg/dl) was two-thirds that observed in the mother's plasma. The normal levels of these apolipoproteins in fetal plasma are well above the sensitivity of the methods, and their quantification requires only 10--20 microliters of fetal plasma. Determination of apolipoproteins in fetal blood obtained by fetoscopy thus may provide a method for the prenatal diagnosis of congenital apolipoprotein deficiencies.

Adult↗
PubMedDate not supplied

Unexplained elevated serum hCG is associated with raised amniotic fluid erythropoietin levels in second-trimester pregnancies.

OBJECTIVE: This study was designed to investigate the association between the concentrations of maternal serum hCG and amniotic fluid erythropoietin during the second trimester of pregnancy. METHODS: In a prospective case-control study, 42 consecutive singleton pregnancies showing unexplained elevated serum hCG concentrations (>2.0 multiples of the median, MoM) in Down's syndrome screening and 27 control pregnant women undergoing midtrimester amniocentesis because of a previous cytogenetic abnormality were studied. RESULTS: The mean amniotic fluid erythropoietin concentration in the study group was 1.8 (range 0.61-8.7) MoM, whereas it was 1.1 (range 0.71-3. 96) MoM in the controls (p = 0.035). A significantly increasing relationship (p < 0.05) was found between the concentrations of maternal serum hCG and amniotic fluid erythropoietin. CONCLUSIONS: The results of the current study revealed in vivo the association between elevated hCG and amniotic fluid erythropoietin levels which, in turn, supports the concept of early placental damage. The underlying pathology seems to be sufficient to cause an erythroblastic response.

Adult↗
PubMedDate not supplied

Ultrasound-detected free-floating particles in amniotic fluid: correlation with maternal serum alpha-fetoprotein.

The objective of this study was to determine the rate of ultrasound-detected free-floating particles in amniotic fluid during the early second trimester and their relationship with maternal serum alpha-fetoprotein (MSAFP) in patients with normal amniotic fluid alpha-fetoprotein (AFAFP). Ninety-eight consecutive patients undergoing second-trimester amniocentesis for various indications were prospectively studied. Before undergoing amniocentesis, each patient had a level II ultrasound examination and evaluation of the presence of free-floating particles. A subjective estimate of the particle amount and measurement of the size of the largest particle seen were made. Patients were stratified into three groups according to their MSAFP level (low, normal, high). Statistical significance of results was assessed by analysis of variance and multiple comparison procedure, and by nonparametric procedures, as appropriate. MSAFPs (mean +/- 1 SD) were 0.41 +/- 0.18 and 4.88 +/- 2.22 multiples of the median for the low and the high groups, respectively. All AFAFPs were within normal limits. Ninety-four percent of patients with high MSAFP had free-floating particles in amniotic fluid as compared to 43% in the low and normal groups (p < 0.01). Patients with high MSAFP had significantly greater density and size of particles. The presence of ultrasound-detected free-floating particles in amniotic fluid of normal patients during the early second trimester may preclude its use as a reliable indicator for fetal lung maturity, or suggest that the source of these particles may differ by trimester. High MSAFP is significantly correlated with the ultrasonographic appearance of free-floating particles, as well as with larger size and higher amount.

Adult↗
PubMed1985

Prostaglandin biosynthesis stimulatory and inhibitory substances in human amniotic fluid during pregnancy and labor.

Human amniotic fluid has been separated into two fractions; one fraction inhibits prostaglandin biosynthesis and the other fraction is stimulatory. The activity of the stimulatory fraction increased with increasing gestational age and was greater still during labor. The activity of the inhibitory fraction decreased with increasing gestational age and was smaller still during labor. We speculate that these changes may play a significant role in parturition.

Amniotic Fluid↗
PubMed1993

Anti-D concentrations in fetal and maternal serum and amniotic fluid in rhesus allo-immunised pregnancies.

OBJECTIVE: To investigate the relation between anti-D concentrations in maternal serum, fetal serum and amniotic fluid, and the development of fetal anaemia. DESIGN: Observational cross sectional and longitudinal study. SETTING: Regional referral centre. SUBJECTS: Sixty-one women undergoing fetal blood sampling at 19 to 36 weeks' gestation for fetal blood and haematocrit estimation for the management of Rh (D) allo-immunisation. Thirty-eight pregnancies (7 with an Rh (D) negative fetus) were tested only once but the rest had two to five fetal blood samplings. INTERVENTIONS: Ultrasound guided fetal blood sampling and amniocentesis, and automated analysis of anti-D antibody quantitation. RESULTS: There were strong correlations between maternal serum, fetal serum and amniotic fluid anti-D concentrations. Analyses of both longitudinal and cross sectional data demonstrated a decrease of the maternal/fetal serum anti-D ratio with gestation. In pregnancies with Rh (D) negative fetuses the maternal/fetal anti-D ratio was significantly lower (P < 0.0001) than in those with Rh (D) positive fetuses. The degree of fetal anaemia (delta haematocrit) was correlated with maternal serum and amniotic fluid anti-D concentrations (r = -0.55, n = 54, P < 0.0001; r = -0.57, n = 44, P < 0.0001, respectively) but there was a weaker correlation with fetal serum anti-D (r = 0.37, n = 54, P < 0.01). CONCLUSION: Anti-D concentrations in maternal serum, fetal serum and amniotic fluid are correlated with fetal anaemia. The decrease in maternal/fetal anti-D ratio with gestation suggests an increase in placental permeability for anti-D with advancing pregnancy.

Amniotic Fluid↗
PubMedDate not supplied

Lack of syringe toxicity to amniotic fluid cells in culture.

Toxicity to amniotic fluid from many brands of syringes has been reported in the past few years. Thirty amniocentesis samples were exposed to Becton-Dickinson syringes for varying lengths of time and evaluated for growth rate. Six amniocentesis samples were exposed to the syringes for prolonged periods at varying temperatures and also evaluated for growth rate. No evidence of syringe toxicity was found for exposure periods up to 24 h, regardless of storage temperature.

Amniotic Fluid↗