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Aseptically collected calf serum as an effective alternative to fetal calf serum in the culture of amniotic fluid cells.

The growth-promoting activities of three different bovine sera have been compared in primary and secondary amniotic fluid cell cultures. In secondary amniotic fluid cell microcultures, aseptically collected calf serum (CS) was slightly, though not significantly more effective than fetal calf serum (FCS) in promoting DNA synthesis, while newborn calf serum (NCS) was significantly less effective than either CS or FCS. All three sera were optimally effective at concentrations of 10 per cent (v/v). Significant variation in quality occurred within four batches of each of CS and FCS, but not within four batches of NCS. A selected batch of CS was significantly more effective in promoting the growth of primary amniotic fluid cell cultures than were a number of batches of FCS then in routine laboratory use. It is suggested that CS may serve as an effective and economical alternative to FCS in the culture of amniotic fluid cells, thereby expanding the scope of serum batch testing. A possible explanation for the varying growth-promoting activities of different sera is discussed.

Amniotic Fluid↗

[Significance of hormones in the amniotic fluid. II. STH and corticoids].

We measured the concentrations of growth hormone and corticosteroids in the amniotic fluid of 50 pregnant diabetic women between the 33rd and 36th week of gestation. These values were compared with normal values obtained by amniocentesis in 275 healthy patients. In the amniotic fluid the concentration of growth hormone decreases during pregnancy. Between the 12th and 16th week we measured 50 ng/ml, whereas in the last four weeks we found 30 ng/ml. In the diabetic patient the amniotic fluid content of STH is lower; however, hyperinsulinaemia of the foetus does not further decrease the concentration of growth hormone. The corticosteroid concentration in the amniotic fluid increases from 10 ng/ml in the 4th month of pregnancy to 35 ng/ml at the end of pregnancy. In diabetic patients we found lower cortisol concentrations; the concentration decreased further in the case of major symptoms of diabetic foetopathy in the newborn. We believe that low corticoid concentration in the amniotic fluid of the diabetic mother may be responsible for the respiratory distress syndrome in the newborn.

Adrenal Cortex Hormones↗

Amniotic fluid volumes in pregnant diabetics during the last trimester. A comparative study using ultrasound and PAH dilution.

Amniotic fluid was measured by ultrasound and PAH dilution technique in 15 insulin-dependent diabetics during the last trimester of pregnancy. Total uterine volume and fetal volume were estimated by ultrasound. The amniotic fluid volume was obtained by subtracting fetal volume and approximated values for placental volume obtained from the literature from the total uterine volume. The mean amniotic fluid volume determined by ultrasound was 1261 ml (range 278-3563). The mean amniotic fluid volume determined by PAH dilution was 1264 ml (range 344-3852). The values for amniotic fluid determined by these two methods were closely correlated (r = 0.96).

Aminohippuric Acids↗

Amniotic fluid embolism with survival.

Well-documented amniotic fluid embolism with survival is an uncommon occurrence. A case is reported with characteristic clinical findings in addition to electrocardiographic evidence of acute right heart strain, disseminated intravascular coagulation, and amniotic fluid debris in central venous blood. The impact of this complication on maternal mortality is emphasized. Aggressive therapy with attention to the basic principles of supportive care offers the best chance of survival.

Adult↗

Morphology of rapidly adhering amniotic-fluid cells as an aid to the diagnosis of neural-tube defects.

In 20 amniotic-fluid samples taken in the second trimester from pregnancies in which the fetus had a neural-tube defect, the proportion of the total viable cell which adhered to glass surfaces after 20 hours in culture ranged from 9 to 100%. In 92 normal amniotic fluids this proportion was less than 6%. Furthermore, the morphology of the rapidly adhering cells was characteristic in spina bifida (8 cases) and anencephaly (12 cases) and distinct from the epithelioid-like cell seen in normal amniotic fluids, including many which were grossly blood-stained. A sample of amniotic fluid from fetal exomphalos and one from a pregnancy in which the placenta had been repeatedly traversed during amniocentesis had proportions of adherent cells in the pathological range, but the morphologies were very different from those seen in neural-tube defects. It is suggested that the techniques described here will be useful adjuncts to amniotic-fluid alpha-fetoprotein determination in the early diagnosis of fetal abnormality, particularly in blood-stained samples.

Adhesiveness↗

Mechanisms by which amniotic-fluid alpha-fetoprotein may be increased in fetal abnormalities.

Assuming that urination is the principle route by which alpha-fetoprotein (A.F.P.) passes into the amniotic fluid from the normal fetus, and that A.F.P. is mainly degraded by a process of fetal swallowing and digestion, 4 possible mechanisms by which amniotic-fluid A.F.P. is increased by fetal abnormalities are postulated.(1) Amniotic-fluid A.F.P. is increased in open neural-tube defects by leakage of A.F.P. from fetal serum and cerebrospinal fluid.(2) In exomphalos exposure of blood-vessels in the extruding viscera permits transudation of A.F.P. into the amniotic fluid.(3) In nephrosis, fetal proteinuria increases amniotic-fluid A.F.P.(4) Impaired fetal swallowing or digestion would account for increased amniotic-fluid A.F.P. in congenital malformations of the orogastrointestinal tract.

Amniotic Fluid↗

Sonographic assessment of amniotic fluid in normal twin pregnancy.

A sonographic standard for assessment of amniotic fluid volume in normal twin pregnancy is provided. In 210 uncomplicated twin pregnancies, amniotic fluid volume was assessed sonographically, by measuring the total amniotic fluid index (AFI), as well as the deepest single vertical pocket in each sac. The mean AFI was greater than values found in singleton gestations; the AFI increased up to 27 weeks' gestation, and decreased thereafter. It correlated well with measurement of the deepest vertical pocket in each sac (r = 0.71; p < 0.0001). In twin pregnancy at 26 to 32 weeks' gestation a fluid pocket of at least 8 cm was found in 9.8% of cases and did not indicate pathologic states. Either AFI or measurement of the deepest fluid pocket in each sac can be used to assess fluid volume in twin pregnancy. A table specific to twin pregnancy should be used when evaluating twin gestations sonographically.

Amniotic Fluid↗

The concentration of the 35-kDa surfactant apoprotein in amniotic fluid from normal and diabetic pregnancies.

A specific, enzyme-linked immunoabsorbent assay was used to determine the concentration of the 35,000 mol wt surfactant apoprotein (SP-A) in samples of amniotic fluid obtained from nondiabetic (n = 358) and diabetic (n = 29) women. The enzyme-linked immunoabsorbent assay was performed with rabbit antibodies directed against SP-A present in lavage fluid from a patient with alveolar proteinosis. Amniotic fluid SP-A concentrations increased as a function of gestational age, from less than 3 micrograms/ml at 30-31 wk to 24 micrograms/ml at 40-41 wk, and were positively correlated with the lecithin to sphingomyelin ratio (p less than 0.01). SP-A concentrations also increased as a function of gestational age in shake test positive samples (p less than 0.05), but were unchanged in shake test-negative samples. There was no difference in the surfactant apoprotein concentration of male compared with female fetuses at any gestational age. In amniotic fluid obtained from 20 diabetic women, SP-A levels were significantly less than in nondiabetic pregnancies that were matched for gestational age and sex of the fetus (p less than 0.05). The SP-A concentrations in amniotic fluids obtained from nine women who were diabetic and hypertensive and from 10 hypertensive women were not different from matched controls. The relationships described above were valid whether the SP-A concentration was expressed per mg protein or per ml amniotic fluid. These data are suggestive that the concentration of amniotic fluid SP-A is decreased in diabetic pregnancies.

Amniotic Fluid↗

Longitudinal measurement of amniotic fluid index in postterm pregnancies and its association with fetal outcome.

OBJECTIVE: Our purpose was to study the association between dynamic changes in serial amniotic fluid index measurements and adverse fetal outcome in postterm pregnancies. STUDY DESIGN: Serial amniotic fluid index values were obtained semiweekly in 139 reliably dated, uncomplicated, singleton pregnancies at > 41 weeks' gestation. Adverse fetal outcome was defined by the presence of moderate or thick meconium, fetal heart rate decelerations, cesarean delivery for fetal distress, neonatal intensive care unit admission, and perinatal mortality. RESULTS: Prominent changes in the amniotic fluid index (i.e., > 50% increase or decrease) had no association with adverse fetal outcome irrespective of rate of change, provided the final value remained > 5.0 cm. A significant association with fetal heart rate decelerations and the presence of meconium was detected only in patients whose final amniotic fluid index was < or = 5.0 cm (p = 0.007 and p = 0.003, respectively). CONCLUSION: Irrespective of the rate of change in amniotic fluid index, postterm pregnancies are significantly associated with potential complications such as fetal heart rate decelerations and meconium staining when the final amniotic fluid index is < or = 5.0 cm.

Amniotic Fluid↗

Phospholipase C activity and phosphatidylinositol in amniotic fluid. A possible contribution of the fetus to the initiation of human parturition.

We investigated the mutual role between amniotic fluid and fetus in the mechanism of initiating human parturition. Phosphatidylinositol in amniotic fluid began to increase from around 30 weeks to 36 or 37 weeks of gestation and then gradually decreased toward term. On the other hand, we demonstrated phospholipase C activity in amniotic fluid, which increased toward term, and we also demonstrated the high phospholipase C activity in neonatal urine, which was 58-fold higher than that in amniotic fluid. The molecular weight of phospholipase C from neonatal urine was estimated to be 33,000 daltons. It was concluded that the fetus relates to onset of labor by producing arachidonic acid in amniotic fluid by the reaction between phosphatidylinositol from fetal lung and phospholipase C from fetal urine.

Amniocentesis↗

[Perinatal outcome in patients with meconial amniotic fluid in labor].

PURPOSE: To evaluate the perinatal morbidity and mortality with the presence of meconial amniotic fluid. MATERIAL AND METHODS: Retrospective study of case review, performed from 1st of June 1995 to May 1997. The patients included were at delivery, with a pregnancy of 32 weeks or older and had meconial amniotic fluid. The variables analyzed were: motherhood age, pre-existing associated illness, resolution of the pregnancy, PSS interpretation, fetal weight, Apgar and final destiny of the product. RESULTS: Of the 432 patients the motherhood aged varied from 13 to 43 years old, mean 27.4; with a number of pregnancies from 1 to 10, mean 2.25. The gestational age went from 32.2 to 42.4 weeks. The fetal weight varied form 1025 to 5080 g. The Apgar grade mean was 7 at the first minute and 8 at the fifth. The pregnancy was interrupted by cesarean in 52.5%. Although there was not a significant difference with the arterial gas, the density of the amniotic fluid did determine the final destiny of the product. CONCLUSIONS: There is a relation between the presence of amniotic fluid and the Apgar grade; both determining the final destiny of the product. When the amniotic fluid had thick meconium the products had a greater morbidity.

Adolescent↗

Gas chromatographic determination with flash methylation of palmitic acid in amniotic fluid in prediction of fetal lung maturity.

A new method for determination of palmitic acid in amniotic fluid was developed. The dipalmitoyl lecithin was hydrolysed, and after extraction the palmitic acid was flash methylated in the injector of the gas chromatograph. The between batch coefficient of variation was 6.0% at 40 and 70 mumol/l of palmitic acid concentrations in 500 microliters amniotic fluid. The accuracy of the method was demonstrated by linearity, recovery and correlation studies. The correlation coefficient was 0.98 between the new and bortrifluoride methylating method. The concentrations of lecithin and palmitic acid in amniotic fluid showed a correlation coefficient of 0.97. If the concentration of palmitic acid falls below 80 mumol/l respiration distress syndrome is likely. The new method was simple and easy to carry out and could be adjusted for haemoglobin contamination of the amniotic fluid. It was concluded that the determination of palmitic acid in amniotic fluid was a reliable method for the assessment of fetal lung maturity that could replace the determination of lecithin concentration.

Amniotic Fluid↗

Biophysical profile with amniotic fluid volume assessments.

OBJECTIVE: To compare the amniotic fluid index (AFI) with the single deepest pocket technique along with the other components of the biophysical profile (BPP) in predicting an adverse pregnancy outcome. METHODS: Prospective, randomized trial of amniotic fluid assessment by AFI or single deepest pocket during a BPP. Cesarean delivery for fetal distress was the primary outcome evaluated. RESULTS: The AFI was used in 273 pregnancies and the single deepest pocket in 264. The AFI significantly increased the number of pregnancies labeled as oligohydramnios, 102 women (38%) compared with 46 women (17%; P <.001), odds ratio (OR) = 2.84, 95% confidence interval 1.90-4.25 in the single deepest pocket group. There was no difference in the number of women with oligohydramnios in the AFI group, 16 of 102 (16%), undergoing a cesarean delivery for fetal intolerance of labor compared with the single deepest pocket group, 6 of 46 (13%; P =.676). More women with normal fluid by the AFI method (AFI > 5), 20 of 170 (12%), underwent a cesarean delivery for fetal distress than the women with normal fluid by the single deepest pocket technique (2 cm x 1 cm pocket present) group, 12 of 218 (6%; P =.037, OR = 2.22, 95% confidence interval 1.05-4.70). CONCLUSION: The AFI offers no advantage in detecting adverse outcomes compared with the single deepest pocket when performed with the BPP. The AFI may cause more interventions by labeling twice as many at-risk pregnancies as having oligohydramnios than with the single deepest pocket technique.

Amniotic Fluid↗

Metabolism of amniotic fluid dopamine by fetal membranes.

The concentrations of L-dopa, three catecholamines and 3,4-dihydroxyphenylacetic acid (DOPAC) were measured both in the amniotic fluid and the fetal urine obtained at term elective cesarean section, using high performance liquid chromatography with electrochemical detection. Dopamine was the major compound in the fetal urine, while DOPAC predominated in the amniotic fluid. Monoamine oxidase (MAO) activity was detected in the fetal membranes by the method of Wurthman and Axelrod. MAO activities in the amnion, chorion and decidua were 0.023 +/- 0.003, 0.180 +/- 0.05 and 2.69 +/- 0.66 nmol/mg protein/min (mean +/- SE), respectively. Incubation experiments, using fetal membranes with human amniotic fluid, demonstrated that dopamine in amniotic fluid was metabolized to DOPAC by fetal membranes. Additional incubation experiments with pargyline, a MAO inhibitor, showed that it strongly inhibited the conversion of dopamine into DOPAC. These results suggest that the dopamine in amniotic fluid was metabolized to DOPAC by MAO in the fetal membranes. This mechanism may regulate the dopamine concentration in amniotic fluid.

3,4-Dihydroxyphenylacetic Acid↗

Detection of proteins in human amniotic fluid using two-dimensional gel electrophoresis.

Samples of human amniotic fluid from 48 pregnancies were examined by high resolution two-dimensional gel electrophoresis and the positions of the major peptides were mapped. Many of the proteins in amniotic fluid also occur in adult and fetal serum. Three regions in the amniotic fluid maps could be defined containing peptides which were not found in adult or fetal serum. The concentration of these peptides is variable and their origin is as yet unknown. Many differences were seen between adult and fetal serum proteins.

Adult↗

Fetal redistribution of blood flow and amniotic fluid volume in growth-retarded fetuses.

Our purpose was to assess the relationship between the fetal redistribution of blood flow and the amount of amniotic fluid in appropriate-for gestational-age fetuses and growth-retarded fetuses. Blood flow velocity waveforms of the umbilical artery, descending aorta, middle cerebral artery, renal artery and uterine artery were recorded using pulsed Doppler ultrasonography in 100 appropriate-for gestational age fetuses and 39 growth-retarded fetuses. The pulsatility index (PI) values and the amount of amniotic fluid were compared between the two groups. The PI values of the umbilical artery and renal artery were significantly higher in appropriate for gestational-age-fetuses with oligohydraminos than in fetuses with an adequate amount of amniotic fluid. The PI values of the umbilical artery and renal artery were significantly higher and the PI of the middle cerebral artery was significantly lower in growth-retarded fetuses with oligohydramnios than in fetuses with an adequate amount of amniotic fluid. Furthermore, there was a significant negative correlation between the PI value of the renal artery and the vertical diameter of amniotic fluid, and between the PI value of the renal artery and the amniotic fluid index. The PI value of the renal artery was related to the amount of amniotic fluid in growth-retarded fetuses, and the same relationship was demonstrated in appropriate-for gestational age fetuses.

Amniotic Fluid↗

Rhesus D genotyping in amniotic fluid.

Rhesus D genotyping by PCR of amniotic fluid offers a very valuable tool for prenatal care of Rhesus D-alloimmunized women. Misleading results have been reported for the method of Bennett et al. analysing the 3'-terminal end of the RhD gene. We have observed also problems with Rhesus D genotyping of DVI-variant samples in the assay of Simsek et al. utilizing size differences of intron 4 between the Rhesus D and the Rhesus CcEe gene. We, thus, prefer to amplify two different regions including the 3'-terminal region of the Rhesus D gene to further minimize the very low risk of false-negative results. This rapid Rhesus D genotyping usually does not require more than 2 ml of amniotic fluid.

Amniocentesis↗

Studies on the elevated amniotic fluid SP1 in Meckel's syndrome: modified glycosylation of SP1.

Pregnancy-specific beta 1-glycoprotein (Schwangerschaftsprotein, SP1) is elevated in the amniotic fluid in Meckel's syndrome. In an attempt to clarify this phenomenon, we studied fetal tissues by immunohistochemistry for the distribution of SP1 in normal fetuses and fetuses with Meckel's syndrome. The amount of SP1 was increased in two out of ten Meckel placentae, as semiquantitatively shown by the immunoperoxidase method, but was normal in the rest of the cases. There was no difference in the SP1 distribution or content in the other fetal tissues studied. Fetal cerebrospinal fluid contained only very low levels of SP1 and thus cannot provide a source for elevated amniotic fluid SP1. In addition, binding of amniotic fluid SP1 to concanavalin A (Con-A) was studied in pregnancies with normal fetuses and Meckel's syndrome using affinity chromatography. The proportion not bound to Con-A in normal amniotic fluids decreased with advancing gestation and was lowest when the total SP1 concentrations were highest. An especially low percentage of SP1 not bound to Con A was found in Meckel's syndrome, suggesting an altered carbohydrate structure of SP1 in these amniotic fluids.

Amniotic Fluid↗