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PubMed1977

Studies on steroid hormone receptors (5alpha-dihidrotesterone, estradiol, and dexamethasone) in cultured human fibroblasts and amniotic fluid cells.

Cultured human fibroblasts and amniotic fluid cells (AF cells) were examined for the presence of steroid hormone receptors. In both cell types, the androgen (DHT) or glucocorticoid (dexamethasone) receptor was detected, but not the estrogen receptor. Binding parameters in fibroblasts were: for androgen: KD = 3.7 X 10(-9) M, Bmax = 13 fmol/mg; for dexamethasone: KD = 4.5 X 10(-9) M, Bmax = 120 fmol/mg. Binding parameters in AF cells were: for androgen: KD = 4 X 10(-9) M, Bmax = 8 fmol/mg; for dexamethasone: KD = 1.9 X 10(-8) M, Bmax = 375 fmol/mg. Cultured cells derived from the gonads (of a patient with 17beta-ketosteroid reductase deficiency) seem to have more receptors than cells from extragenital body parts (Bmax = 21 fmol). With the aid of gel chromatography, the molecular weight of the androgen receptor was estimated to be 30-40 000D.

Amniotic Fluid↗
PubMed1996

Prenatal diagnosis of organic acidemias based on amniotic fluid levels of acylcarnitines.

Acylcarnitines in amniotic fluid samples were analyzed for the prenatal diagnosis of propionic acidemia, methylmalonic aciduria, isovaleric acidemia, and glutaric aciduria by electrospray tandem mass spectrometry. Although the levels of the specific acylcarnitine between affected and unaffected cases showed an overlap, the ratios of propionylcarnitine to 4-carbon acylcarnitine levels for propionic acidemia and methylmalonic aciduria, those of isovalerylcarnitine to propionylcarnitine for isovaleric acidemia, and those of glutarylcarnitine to propionylcarnitine for glutaric aciduria type I were shown to be reliable indicators in the prenatal diagnosis. In addition, it is suggested that the combination of the ratios of glutarylcarnitine, isovaleryl-carnitine, and hexanoylcarnitine to propionylcarnitine may be useful for the prenatal diagnosis of glutaric aciduria type II.

Acetylcarnitine↗
PubMed1979

[Method of culturing amniotic fluid cells].

The influence of pregnancy term and the concentration of cell suspension, introduced into culture, on the growth of amniotic fluid cells have been studied on 29 specimens. Optimum term of amniotic fluid cells sampling for the best growth was 17 weeks of pregnancy. It is recommended to take into account the concentration of cell suspension in the amniotic fluid before introducing the cells into the culture depending on pregnancy terms.

Amniotic Fluid↗
PubMed1976

Partial characterization of somatomedin bioactivity in term human amniotic fluid.

Somatomedin (SM) activity in human amniotic fluid (AF) was partially characterized. Aliquots of pooled AF, obtained at term gestation, were acidified to pH 2.3 and sequentially studied by ultrafiltration on various membranes, Sephadex gel filtration, and starch gel electrophoresis (SGE). SM activity was measured by a rat cartilage bioassay. SM activity in AF, under these conditions, was present in several stable indicated molecular size (IMS) forms; large (greater than 45,000) and small (near 10,000). The small SM was found in basic and acidic forms after SGE. These studies suggest that SM activity in term human AF is somewhat similar to SM in unextracted plasma. In addition, immunoreactive GH and insulin were measured in AF but at lower concentrations than usually found in plasma. After chromatography of AF, GH was found in large and small IMS forms, whereas most of the insulin was found in the monomer insulin IMS area.

Amniotic Fluid↗
PubMed1980

Phospholipid studies of the amniotic fluid in diabetic pregnancy.

The lecithin/sphingomyelin ration (L/S) of the amniotic fluid is a reliable indicator of fetal lung maturity in most disorders complicating pregnancy. False positive results of L/S occur especially in diabetic pregnancies. Phosphatidylglycerol (PG) improves the ability of the surface active lecithin to prevent lung atelectasis. Respiratory distress syndrome of the newborn infant does not seem to occur when PG is present. By determining PG of the amniotic fluid, false positive L/S can be excluded in diabetic pregnancies.

Amniotic Fluid↗
PubMed1989

Detection of bisected biantennary form in the asparagine-linked oligosaccharides of fibronectin isolated from human term amniotic fluid.

Fibronectin purified from human term amniotic fluid contains 10 asparagine-linked sugar chains in one molecule. The sugar chains were quantitatively liberated as radioactive oligosaccharides from the polypeptide moiety by hydrazinolysis followed by N-acetylation and NaB3H4 reduction and fractionated by anion-exchange column chromatography and serial lectin affinity chromatography. The structures of these sugar chains were determined by sequential exoglycosidase digestion in combination with methylation analysis. The results indicated that they are a mixture of bisected and non-bisected bi- and triantennary complex-type sugar chains with and without a fucose on the proximal N-acetylglucosamine residue and with Gal beta 1----4GlcNAc beta 1----, GlcNAc beta 1----, Neu5Ac alpha 2----3Gal beta 1----4GlcNAc beta 1----, and Neu5Ac alpha 2----6Gal beta 1----4GlcNAc beta 1---- groups in their outer chain moieties.

Amniotic Fluid↗
PubMed1983

Profile of phospholipids in amniotic fluid to assess fetal lung maturity in high-risk pregnancy.

In assessing fetal lung maturity, an amniotic fluid profile, consisting of a total phospholipid (as phosphorus) (TPP) and total lecithin (LT) concentration, has been shown to yield a predictive accuracy of 97.5% with a false-negative rate of 2.4%. The LT had a range of specificity of 93% to 99%, whereas the one for the TPP was 88% to 96%. The cut-off values were 0.14 mg P/dl and 3.5 mg lecithin/dl of amniotic fluid. The analytical variables for laboratory tests for measuring amniotic fluid phospholipids led to the development of the quantitative tests used for this study. The proposed profile appears to offer the clinician an excellent means of judging how to manage the high-risk pregnancy.

Amniocentesis↗
PubMed1991

[Immunological properties of the amniotic fluid in physiological pregnancy and gestoses].

Immunologic analysis of the amniotic fluid included measurements of immunoglobulins A and M, circulating immune complexes, rheumatoid and antinuclear factors, immunophoresis and electrophoresis, and tests for the presence of donor lymphocytes, Changed immunologic parameters of the amniotic fluid indicate a complicated course of pregnancy, development of gestosis among other things. Basing on their findings, the authors suggest that gestosis involve dysfunction of the adaptive mechanisms, that may result from disordered interactions of the immunity system components.

Adolescent↗
PubMed1991

Amniotic-fluid ingestion enhances morphine analgesia during morphine tolerance and withdrawal in rats.

Ingestion of placenta and amniotic fluid has been shown to enhance opioid-mediated analgesia in rats produced by morphine injection, footshock, vaginal/cervical stimulation, and during late pregnancy. The present study was designed to investigate the effects of amniotic fluid ingestion on the characteristics of morphine dependency and withdrawal. Tail-flick latencies in Long-Evans rats were determined before and after repeated daily injections of morphine sulfate. It was found that ingestion of amniotic fluid after establishment of the morphine dependency, coupled with an injection of an otherwise ineffective dose of morphine, enhanced analgesia in morphine-dependent rats, and reversed hyperalgesia seen during withdrawal from morphine dependency.

Amniotic Fluid↗
PubMed2006

[Origin and significance of leptin and neuropeptide Y (NPY) in amniotic fluid between the 14th and 18th weeks of gestation].

INTRODUCTION: Leptin and neuropeptide Y (NPY) play important roles in the regulation of food intake, energy expenditure, hematopoesis and reproduction. The biological functions of leptin and NPY in fetal development, and their regulation during pregnancy by fetal and maternal factors remain poorly understood. PATIENTS AND METHODS: From 55 women undergoing diagnostic amniocentesis between the 14th and 18th weeks of gestation samples of amniotic fluid were collected. In accord with the circadian rhythm of leptin secretion all amniocenteses were performed between 8.00 and 12.00 a. m. The concentrations of leptin and NPY in amniotic fluid were analysed using commercially available RIA's. RESULTS: The amniotic fluid samples of 32 male and 23 female fetuses were determined and demonstrated no gender-dependent differences in leptin and NPY levels. No correlation was found between leptin/NPY and the maternal body mass index. NPY concentrations are lower in advanced gestational age pregnancies. Leptin levels revealed no differences with respect to gestational age. CONCLUSION: Leptin and NPY levels were independent of fetal gender and maternal BMI. This provides clues for alternative regulatory mechanisms in leptin and NPY secretion. Compared to the data of adults, our findings show high leptin concentrations in the amniotic fluid in the presence of only minor amounts of white adipose tissues which might include the placenta. Our data including the observation of lower levels of NPY in more advanced gestational age support the idea of specific factors regulating leptin and NPY secretion into the amniotic fluid.

Adult↗
PubMed1998

Amnioinfusion in labor induction of term pregnancies with premature rupture of the membranes and low amniotic fluid.

OBJECTIVE: To analyze the utility of prophylactic amnioinfusion in term pregnancies with PROM and a low amniotic fluid index during labor induction. METHOD: Forty-two women with amnioinfusion and 42 in a control group with amniotic fluid index (AFI) below 10 cm when admitted to labor induction were studied. All patients had electronic fetal heart rate and intrauterine pressure continuous monitoring. Amnioinfusion of normal saline (37 degrees C) was realized in the study group, using a continuous perfusion pump at 600 ml/h for 1 h, after which the AFI was again recorded; if this was < 15, the perfusion was continued at 180 ml/h until full cervical dilatation was achieved or until uterine baseline activity reached 20 mm Hg. The control group received identical obstetric care except in respect of amnioinfusion. RESULT: Both groups were similar in age, primiparity, gestational age, initial AFI, interval from rupture of membranes until delivery and length of labor. The amnioinfusion of 600 ml in 1 h significantly increased the AFI (an increase of 7.2 +/- 3.9 vs. a decrease of 1.1 +/- 1.6, P < 0.01). In the amnioinfusion group, there was a significantly lower rate of cesarean deliveries (0 vs. 6, P < 0.05) and a better mean umbilical arterial pH at delivery (7.24 +/- 0.07 vs. 7.21 +/- 0.08, P < 0.05). No differences were observed in maternal or neonatal hospitalization days or infectious morbidity. CONCLUSION: It is concluded that prophylactic amnioinfusion improves neonatal metabolic state when used in labor induction of term pregnancies with PROM and a low amniotic fluid index.

Adult↗
PubMed1981

Diagnosis of neural tube defects by estimation of amniotic fluid acetylcholinesterase.

Acetylcholinesterase (AChE) activity in amniotic fluid was assayed directly by a reaction rate method at 30 degrees C using acetylthiocholine iodide as substrate and ethopropazine as a safe 'pseudo' cholinesterase inhibitor. Fresh samples from 101 normal pregnancies of 14-24 weeks gestation had a mean AChE activity of 2.56 u/l (SD 1.10). Elevated levels of AChE were found in association with open spina bifida (5.5-20.4 u/l), anencephaly (10.2-19.5 u/l), exomphalos (2.7-15.6 u/l), intrauterine death (30.2-59.3 u/l) and Turner's syndrome (36.4 u/l) but not with closed spina bifida or rhesus isoimmunisation. When AChE activity was expressed as a percentage of the total cholinesterase activity ('percentage AChE'), there was a good correlation between AChE activity and 'percentage AChE' in normal pregnancies and the values associated with the 37 pregnancies affected by open neural tube defects (NTD) fell outside the 99.9 per cent confidence limits of the normal group. Qualitative differences in cholinesterase activity could be demonstrated between the groups with exomphalos and open NTD. It is suggested that the assay might be used satisfactorily to demonstrate the presence of open NTD in affected pregnancies.

Acetylcholinesterase↗
PubMed1997

Open neural tube defects: immunocytochemical demonstration of neuroepithelial cells in amniotic fluid.

Cytologic evaluation of second trimester amniotic fluid (AF) is a rapid, inexpensive adjunct to prenatal diagnosis of open neural tube defects (ONTDs). Our goal was to determine whether the neural-appearing cells and/or large foamy macrophages in the AF of anencephalics are indeed of neural and/or glial origin. In two second trimester patients with elevated serum alpha-fetoprotein (AFP) and polyhydramnios, fetal sonogram studies showed anencephaly; amniocentesis was performed for AF-AFP, cytogenetic, and cytologic studies. AF sediment smears were initially Papanicolaou-stained; next, the same smears were immunoperoxidase (IP)-stained for glial fibrillary acidic protein (GFAP). If GFAP negative, slides were restained for synaptophysin (SYN) and neuron-specific enolase (NSE). Both AFs contained small neural-appearing cells (5-10 microns) singly and in clusters, with dense, round, homogeneous nuclei, an occasional nucleolus, and scant cytoplasmic rim. These were GFAP negative and SYN and NSE positive; the large vacuolated, lipid-laden macrophages (20-40 microns) were negative for all three IP stains. In conclusion, positive IP staining for SYN and NSE supports the morphologic impression that small dark cells in AF are of neural origin, while negative IP staining of large foamy macrophages suggests nonneural, nonglial origin.

Amniotic Fluid↗
PubMed1983

Differences in the carbohydrate moieties of plasma and amniotic fluid fibronectins as revealed by crossed immunoaffinity electrophoresis with concanavalin A.

Human fibronectins isolated from pooled human plasma and amniotic fluid were studied as to differences in their carbohydrate moieties. The chemical analyses showed that amniotic fluid fibronectin is different from adult plasma fibronectin in carbohydrate content and composition while there seems to be no significant differences in amino acid composition. Crossed immunoaffinity electrophoresis with free concanavalin A, as well as rocket immunoelectrophoresis with immobilized concanavalin A intermediate gel, indicated that amniotic fluid fibronectin has little or no reactivity with this lectin while adult plasma fibronectin is strongly reactive. Fetal cord plasma fibronectin apparently interacted with concanavalin A, but its reactivity was weaker than that of adult plasma fibronectin. Fibronectin isolated from ascitic fluid of an ovarian cancer patient which was examined in additional experiments showed much weaker Con A-reactivity than fetal cord plasma fibronectin. These results suggest that fibronectins from various body fluids differ in their carbohydrate structures.

Amino Acids↗
PubMed2002

Interleukin-6 and interleukin-1 receptor antagonist in amniotic fluid and cord blood in patients with pre-term, premature rupture of the membranes.

OBJECTIVE: We investigated the role of interleukin (IL)-6 and IL-1 receptor antagonist (IL-1ra) in pre-term premature rupture of the membranes (PROM). METHOD: Amniotic fluid samples were collected from 10 patients with pre-term PROM (group 1a), 13 patients undergoing genetic amniocentesis (group 2), seven patients with normal vaginal delivery (group 3a), and 11 patients with elective cesarean section (group 4a). Umbilical venous blood was collected from nine cases of pre-term PROM (group 1b), 19 cases of normal delivery (group 3b) and nine cases of elective cesarean section (group 4b). RESULTS: The concentration of IL-6 in the amniotic fluid in group 1a was significantly higher than in group 2 (P<0.001), and that in group 3a was significantly higher than in group 4a (P<0.001). The concentration of IL-1ra in amniotic fluid in group 1a was significantly higher than in group 2 (P<0.001). The concentrations of IL-6 in umbilical venous blood were not significant among the groups. The concentration of IL-1ra in umbilical venous blood in group 1b was significantly higher than in group 3b (P<0.05) and group 4b (P<0.05). CONCLUSIONS: High concentrations of amniotic fluid IL-6 present in pre-term PROM, are indicative of intrauterine inflammation, probably due to sub-clinical infection. The high concentrations of IL-1ra in amniotic fluid and umbilical venous blood suggest the possibility that the presence of this anti-inflammatory cytokine serves to prevent the development of overt inflammation.

Amniotic Fluid↗
PubMed2003

Effect of adeno-associated virus-specific immunoglobulin G in human amniotic fluid on gene transfer.

Intra-amniotic administration of adeno-associated virus (AAV) vector may be an effective way to deliver gene therapy for treatment of congenital pulmonary and intestinal disorders. In an effort to understand potential barriers to intra-amniotic gene therapy better, we determined whether human amniotic fluid (AF) could act as an inhibitor of AAV2-mediated gene transfer. AF samples were obtained from 21 different human pregnancies during routine amniocentesis at 16-20 weeks of gestation. An immortalized fetal human tracheal epithelial cell line (FHTE) was infected with AAV2 containing a luciferase reporter gene driven by the SV40 promoter in the presence and absence of each AF sample. Inhibition of transgene expression was observed in 8 (38%) of the AF samples (inhibitory AF) and resulted in luciferase levels of only 1.4% +/- 0.6% of those obtained with infection in normal media. Infections in 13 samples (62%) resulted in transgene expression comparable or in excess of infection in media alone (noninhibitory AF). Removal of immunoglobulin G (IgG) from inhibitory AF samples with Protein A returned luciferase expression to control levels (119% +/- 37% of control), suggesting the possible presence of inhibiting antibody. Eleven of the AF samples were evaluated by enzyme-linked immunosorbent assay (ELISA) for specific anti-AAV antibodies. All noninhibitory AF samples were negative (titers of < 1:20; n = 3), and 6 of the 8 inhibitory samples contained specific anti-AAV antibodies at titers ranging from 1:40 to 1:160. These studies demonstrate that AF from some individuals contains AAV-specific IgG that can inhibit gene transfer.

Amniotic Fluid↗
PubMed1986

Trichloroacetic acid accumulates in murine amniotic fluid after tri- and tetrachloroethylene inhalation.

The distribution of trichloroethylene (Tri) and tetrachloroethylene (Tetra) and their metabolites have been studied in pregnant mice by means of whole-body autoradiography (14C-labelled Tri and Tetra) and gas chromatography, with special emphasis on possible uptake and retention in the foetoplacental unit. Volatile (non-metabolized) activity appeared at short intervals after a 10 min. or 1 hr inhalation period in foetus and amniotic fluid. Most notable, however, was a strong accumulation and retention (peak at 4 hrs) in amniotic fluid of the metabolite trichloroacetic acid (TCA) after inhalation of either of the solvents. The main metabolite of Tri, trichloroethanol (TCE) (or conjugates), did not accumulate specifically as compared to maternal plasma. TCA infused intravenously in the maternal plasma was accumulated in amniotic fluid, but less pronounced than after Tri and Tetra inhalation, indicating that some metabolism of Tri and Tetra to TCA may occur in the foetoplacental unit. The results suggest that TCA may be transported to the foetus partly paraplacentally through foetal membranes and amniotic fluid, with the possibility of foetal swallowing or absorption through the skin. Foetal urinary activity also suggests that circulation between foetus and amniotic fluid may contribute to the long-term retention in the foetoplacental unit. In the mother, after inhalation exposures, and in intraperitoneally injected newborn mice, non-extractable radioactivity was found in the respiratory tract, liver, and kidney, indicating binding to these organs through metabolism.

Amniotic Fluid↗
PubMed2002

Decreased laboratory testing for lecithin-to-sphingomyelin ratio and phosphatidylglycerol after fetal lung maturity assessment from lamellar body count in amniotic fluid.

The objectives of this study were to do inexpensive lamellar body count (LBC) in amniotic fluid, to do statistical analysis to evaluate cutoff values for fetal lung maturity (FLM) and fetal lung immaturity (FLI), to derive a threshold for obtaining a lecithin-to-sphingomyelin (L/S) ratio and phosphatidylglycerol percentage (%PG), and to determine the potential cost savings to the hospital if they use this new method. Testing (LBC, L/S ratio, and %PG) was done on 123 specimens of amniotic fluid. Receiver operating characteristic (ROC) curve, discriminant, linear regression, chi2, and cost analyses were used to evaluate the laboratory and financial parameters. Lamellar body counts of greater than 41,500 (Coulter MAXM: sensitivity, 90.5%; specificity, 87.7%; positive predictive value, 79.2%; negative predictive value, 94.7%) and greater than 32,000 (Coulter Gen.S: sensitivity, 90.5%; specificity, 85.2%; positive predictive value, 76.0%; negative predictive value, 94.5%) were the best threshold for biochemical FLM. Similarly, LBC of less than 24,000 (MAXM: sensitivity, 78.6%; specificity, 100%; positive predictive value, 100%; negative predictive value, 90.0%) and less than 21,000 (Gen.S: sensitivity, 71.4%; specificity, 100%; positive predictive value, 100%; negative predictive value, 87.1%) provided the best statistical cutoff for biochemical FLI from discriminant analysis. The authors concluded that FLM and FLI can be predicted with reasonable accuracy from LBC in amniotic fluid specimens. The expensive and not easily accessible L/S ratio and %PG can then be done only in cases in which LBC indicates transitional FLM. A cascade approach results in 86% savings to the hospital if the L/S ratio and %PG are not sent to a reference laboratory.

Adult↗