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[Radioimmunological determination of insulin in the amniotic fluid].

Small amounts of immunoreactive insulin, at about one-eighth the concentrations found in maternal serum, have been detected in human amniotic fluid. A positive correlation between term amniotic fluid insulin levels and newborn birth weight was found. Insulin was studied in 42 samples of amniotic fluid obtained from normal and pathological pregnancies by radioimmunoassay. In only one sample of amniotic fluid of diabetic mother with insulin dependence we found high values of insulin and condition of insufficient fetal lung maturity. We believe that radioimmunoassay of insulin in amniotic fluid may be a parameter of evaluation of fetal maturity.

Adult↗

Immunoreactive endorphin peptides in amniotic fluid during labour.

Immunoreactive endorphin peptides (irEP) were measured in term amniotic fluids at induction by amniotomy (n = 8), during early and late first stage of labour (n = 52), at vaginal delivery (n = 30) and at caesarean section (n = 10). Individual values ranged from 50 to 222 pg/ml, but mean values did not differ significantly, and serial determinations revealed that irEP in the amniotic fluid remained almost constant in each woman from the first stage of labour through to delivery. There was no correlation between pathological changes in the cardiotocogram or low umbilical artery pH and irEP levels in the amniotic fluid at vaginal delivery. We conclude that immunoreactive endorphin peptides in the amniotic fluid probably are of fetal or placental origin and do not reflect fetal wellbeing or distress.

Amniotic Fluid↗

ED-A sequence containing fibronectin in human amniotic fluid and amnion epithelial cells.

1. Human amniotic fluid fibronectin (aFn) was studied by using a monoclonal antibody 52DHl (DH) that recognizes the extra domain (ED-A) sequence of cellular Fn (cFn). 2. In immunoblotting the DH antibody reacted with a sharp polypeptide band at the top of the bulk of the diffuse aFn. Another monoclonal antibody 52BF12 (BF) against the cell binding site of Fn, recognized the whole aFn. 3. The ED-A sequence containing cFn (EcFn) formed a constant proportion in aFns from all amniotic fluid preparations studied. 4. In amniotic membranes the DH antibody revealed bright subepithelial immunofluorescence. 5. Also isolated and cultured human amnion epithelial cells were strongly positive in immunofluorescence and secreted EcFn into the culture medium as revealed by immunoblotting. 6. The results indicate that aFn is a composition of at least two different Fn subtypes of which the EcFn most probably originates from amnion epithelial cells.

Amnion↗

gamma-Glutamyl transpeptidase of human amniotic fluid.

gamma-Glutamyl transpeptidase (GGTP) activity in normal amniotic fluids and corresponding maternal sera obtained at various gestational periods was measured. The ontogenic pattern of enzyme activity in amniotic fluid is very similar to alpha fetoprotein (AFP). However, the levels of these two proteins behaved differently in corresponding maternal sera. Also, in amniotic fluids obtained from pregnancies with neural tube defects (NTD), only AFP concentration was abnormally high whereas GGTP activity was normal.

Amniotic Fluid↗

Normal values for human umbilical venous and amniotic fluid pressures and their alteration by fetal disease.

Umbilical venous and amniotic fluid pressures were measured in 68 human pregnancies at the time that cordocentesis was performed. Normal umbilical venous pressure was unrelated to gestational age and remained within a tight range (5.3 +/- 2.3 mm Hg, mean +/- SD). Fetuses with an elevated umbilical venous pressure had disorders consistent with either hepatomegaly or congestive heart failure. Umbilical venous pressure was significantly increased before treatment in two fetuses with immune hydrops; it rapidly declined with treatment. Neither gestational age nor umbilical venous pressure was significantly different in the groups that received and did not receive pancuronium. There was a strong relationship between amniotic fluid pressure and gestational age in normal pregnancy (r = 0.54, p less than 0.0001). Women with hydramnios had amniotic fluid pressures greater than control subjects (p = 0.0007). This investigation documents normal human amniotic fluid and fetal umbilical venous pressures. These measurements are altered by disease and may prove to be of diagnostic and therapeutic value in the future.

Amniotic Fluid↗

Accuracy of portable glucose meters for rapid determination of amniotic fluid glucose levels.

This study was designed to evaluate the accuracy and feasibility of use of three commercially available portable blood glucose meters to measure amniotic fluid glucose(AFG) levels as compared to an accepted laboratory standard. A prospective study of amniotic fluid from 101 consecutive amniocenteses was performed. Glucose concentration in the amniotic fluid was assessed by hexokinase method in our hospital laboratory (control) and by using three portable meters: Advantage (ADV) (Boehringer Mannheim), Glucometer Elite (ELT) (Bayer), and One Touch II Hospital (T-2) (Lifescan). Twenty consecutive amniotic fluid samples were sent to the laboratory in two vials, the first without additive and the second with potassium oxalate to prevent metabolic activity, to assess the effect of cellular metabolism and time delay on amniotic fluid glucose concentrations. Data are reported as mean +/-SE and were assessed by one-way ANOVA. Of the 101 patients studied, 29 were of gestational age > or = 20 wks. The remaining 72 patients were < 20 wks. All three ambulatory meters demonstrated a linear relationship with control (all P < 0.001). Given a slope of almost 1.0 (m = 0.94) and a y-intercept approaching zero (b = 4.3), the OT2 proved to correlate best with control. ELT: (r2 = 0.55, m = 0.79, b = 22.2) and ADV: (r2 = 0.74, m = 1.45, b = 16.9) both overestimated amniotic fluid glucose. When AFG was < 30 mg/dl via laboratory standard, OT2: (r2 = 0.78, m = 1.05, and b = -2.20, P < 0.001), ADV: (m = 1.02, b = 24.1, r2 = 0.12, P = 0.133). The One Touch II Hospital accurately predicted amniotic fluid glucose at the bedside with excellent correlation including with laboratory standard glucose levels < 30 mg/dl. ADV and ELT proved too inaccurate for clinical use. Control samples were not affected by additives or time delay. These findings confirm that AFG determinations can be obtained rapidly with the OT2 meter at the bedside.

Adult↗

Function curve of the membranes that regulate amniotic fluid volume in sheep.

Seven singleton 120-day fetal lambs were prepared with a shunt from the lung to the gastric end of the esophagus, a bladder catheter, and multiple amniotic fluid and vascular catheters. The urachus was ligated. Beginning 7 days later, amniotic fluid volumes were determined by drainage, followed by replacement with 1 liter of lactated Ringer (LR) solution. Urine flow into the amnion was measured continuously. In 14 of 27 experiments, amniotic fluid volumes were determined again 2 days after the inflow into the amnion had consisted of urine only and in 13 experiments after the inflow of urine had been supplemented by an intraamniotic infusion of LR solution. Intramembranous absorption was calculated from the inflows and the changes in volume between the beginning and end of each experiment. The relations between absorption rate and amniotic fluid volume, the "function curves," were highly individual. Urine production during the infusion of LR solution did not decrease, fetal plasma renin activity decreased (P < 0.001), and amniotic fluid volume increased by 140% [SE (27%), P < 0.005], but the increase in the amniochorionic absorption rate of 411% [SE (48%), P < 0.001] was greater (P < 0.005) than the increase in volume. Each of the seven fetuses was proven capable of an average intramembranous absorption rate that exceeded 4.5 liters of amniotic fluid per day. During the infusion of LR solution, the increase in the rate of absorption matched the rate of infusion (both in ml/h), with a regression coefficient of 0.75 (P < 0.001). Thus, even for large amniotic fluid volumes, volume is not limited by the absorptive capacity of the amniochorion, and, at least in these preparations, the position of the function curve and not the natural rate of inflow was the major determinant of resting amniotic fluid volume.

Absorption↗

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.

Adult↗

The relationship between trypsin activity in amniotic fluid and premature rupture of membranes.

Trypsin activity and concentration of alpha 1-antitrypsin in amniotic fluid were measured in patients with and without premature rupture of membranes. Trypsin activity in amniotic fluid with premature rupture of membranes (127 +/- 57 U/L) was found to be significantly higher than in cases without premature rupture of membranes (14 +/- 20 U/L) (p less than 0.01). The concentration of alpha 1-antitrypsin in amniotic fluid with premature rupture of membranes (6.8 +/- 5.5 mg/dl) was significantly lower than in cases without premature rupture (25.2 +/- 20.2 mg/dl) (p less than 0.01). Trypsin activity and concentration of alpha 1-antitrypsin in amniotic fluid were not influenced by rupture of membranes and labor. These studies suggest that high trypsin activity and low concentration of alpha 1-antitrypsin in amniotic fluid is one cause of premature rupture of membranes.

Amnion↗

Amniotic fluid matrix metalloproteinase-8 and the development of cerebral palsy.

AIMS: To examine if increased concentrations of matrix metalloproteinase-8 (MMP-8) in amniotic fluid are associated with the development of cerebral palsy at the age of three years. METHODS: The relationship between amniotic fluid concentrations of MMP-8 and the development of cerebral palsy was examined in 116 preterm singleton newborns (gestational age at birth < 35 weeks) born to mothers who underwent amniocentesis and were followed for at least 3 years. Amniotic fluid was cultured for aerobic and anaerobic bacteria and mycoplasmas. MMP-8 concentrations were measured by specific immunoassays. Cerebral palsy was diagnosed by neuro developmental assessment at the age of three years. RESULTS: Median amniotic fluid concentration of MMP-8 was significantly higher in mothers whose newborns developed cerebral palsy than in mothers whose newborns did not develop cerebral palsy (median 153.9 [range < 0.3-1535.9] ng/ml vs median 6.4 [range < 0.3-3836.8] ng/ml; p < 0.01). Neonates who developed cerebral palsy were delivered at earlier gestational age than those without cerebral palsy. After adjustment for the gestational age at birth and the results of amniotic fluid culture, elevated concentrations of amniotic fluid MMP-8 significantly increased the odds of development of cerebral palsy (odds ratio, 6.0; 95% confidence interval, 1.1-33.0; p < 0.05). CONCLUSION: Increased concentrations of amniotic fluid MMP-8 are associated with the subsequent development of cerebral palsy at the age of 3 years.

Adult↗

Spectrophotometric determination of meconium concentration in amniotic fluid.

When a fetus is acutely distressed, for example as a result of strangulation by the umbilical cord, the bowel content, meconium, may be passed into the amniotic fluid. Assessment of meconial contamination of the amniotic fluid is important in the management of late pregnancy. Fresh or increasing meconium content is an indication for urgent delivery. 'Old' meconium is usually distinguished from 'new' by visual inspection of the stained amniotic fluid; a more objective method could have advantages. We consider the effects of meconium staining of amniotic fluid on the spectrophotometric properties of the liquor. The absorption spectra of meconial amniotic fluids reveal an excess absorption band, centred around 410 nm. Quantification of the band height is proposed and it is shown that the height is linearly related to meconium concentration, with a proportionality factor, or sensitivity, of 0.046 +/- 0.009 (mean +/- s.d.) proportionality factor, or sensitivity, of 0.046 +/- 0.009 (mean +/- s.d.) optical density (OD) units per g l-1. This sensitivity factor may be used to estimate meconium concentration in samples of amniotic fluid.

Amniotic Fluid↗

Cytologic and biochemical changes associated with inoculation of amniotic fluid and meconium into lungs of neonatal rats.

OBJECTIVE: To evaluate the effect of homologous amniotic fluid and meconium inoculated intratracheally into the lungs of neonatal rats. ANIMALS: 153 male 7-day-old Fischer-344 rats. PROCEDURE: Amniotic fluid was obtained by cesarean section from the uterus of pregnant rats and meconium was collected at the time of birth from the gastrointestinal tract of neonatal rats. Neonatal rats were randomly allocated into 5 treatment groups. Two groups received 0.05 ml of saline (0.9% NaCl) solution; the third and fourth groups received 0.05 ml of 50% or 100% amniotic fluid, respectively; the fifth group was inoculated with 0.05 ml of a 20% suspension of meconium. Six or 7 rat pups/group were euthanatized by exsanguination under halothane anesthesia at postinoculation days 1, 3, 7, and 14. The magnitude of injury and inflammatory response was determined by biochemical and cytologic analyses of bronchoalveolar lavage fluid. RESULTS: Inoculation with saline solution and amniotic fluid did not induce pulmonary injury or inflammatory response. Inoculation with meconium induced significant (P < 0.01) injury and inflammatory response, characterized by the release of cytosolic enzymes and recruitment of neutrophils in the lung. CONCLUSIONS: Saline solution is an innocuous vehicle that can be safely used in intratracheal inoculations in neonatal rats. Homologous amniotic fluid, despite containing keratin and epidermal cells, does not cause acute injury or inflammation in the lung. In contrast, meconium acts as a toxic substance injuring respiratory cells and causing a vigorous but transient leukocytic inflammatory reaction in the lungs.

Alkaline Phosphatase↗

[The value of various investigations of amniotic fluid in the estimation of fetal maturity (author's transl)].

The amniotic fluid lecithin-spingomyelin (L/S) ratio, creatinine and uric acid concentrations, percentage of organe-stained fat cells and the foam test by Clements and coworkers have been compared in 82 samples of amniotic fluid from 66 patients. The specimens were obtained by transabdominal amniocentesis or amniotomy between the 24th and the 42nd week of pregnancy. The determination of the L/S ratio and the foam test seem to be reliable methods of estimating pulmonary surfactant and, hence, of predicting the respiratory distress syndrome (RDS) in the newborn infant. There was no RDS with an L/S ratio greater than 1.6 to 1.8. Positive foam tests also seem to be a valuable indicator of pulmonary maturity, no cases of RDS being found. However, false negative foam tests are not rare. The amniotic fluid concentration of creatinine correlated well with gestational age and birth weight. The determination of uric acid in the amniotic fluid is an unreliable test of fetal maturity on account of the large scatter. The percentage of orange-stained cells did not rise above 10% before the 39th week of pregnancy in most cases.

Amniotic Fluid↗

Sonographic amniotic fluid measurement and fetal growth retardation: a reappraisal.

Sonographic estimation of amniotic fluid volume is commonly used as a diagnostic test for oligohydramnios associated with fetal malnutrition. However, methods of measurement and diagnostic criteria are controversial. We prospectively evaluated the reproducibility of amniotic fluid diameter measurement and studied various threshold definitions of oligohydramnios as predictors of small for gestational age fetuses or fetuses with low ponderal index values. An average of the vertical and two perpendicular horizontal diameters of the largest pocket of amniotic fluid was more reproducible than the maximum vertical pocket and better enabled a single observer to differentiate between patients. An average diameter of 10 mm was highly diagnostic of small for gestational age fetuses but was insufficiently sensitive (27%) to serve as a screening test. Although also of limited sensitivity (40%), an average diameter of approximately 30 mm was probably the best screening threshold. Reduced average diameter was significantly related to the occurrence of a ponderal index less than or equal to the tenth percentile for dates. Three of seven (43%) neonates with a ponderal index less than or equal to the tenth percentile and a birth weight greater than the tenth percentile for dates had a low average diameter, suggesting that reduced average amniotic fluid diameter may help to identify the malnourished fetus with birth weight greater than the tenth percentile for dates.

Amniotic Fluid↗

[Metabolic activation of benzo(a)pyrene by human amniotic fluid].

The in vitro activation of benzo(a)pyrene was studied in amniotic fluid from ten 4-month pregnant women. Benzo(a)pyrene monooxygenase and epoxide hydrolase activities were in the same range in amniotic fluid as in human liver. Glutathione epoxide transferase activity was markedly lower than in hepatocytes. Human amniotic fluid also catalyzed the formation of hydrocarbon metabolites mutagenic to Salmonella typhimurium TA98 (Ames system). Profiles of amniotic fluid aromatic hydrocarbons from non smokers exhibited low benzo(a)pyrene concentration (less than 0.1 ng/ml).

Amniotic Fluid↗

Amniotic fluid testosterone and follicle-stimulating hormone in the determination of fetal sex.

To determine whether hormone analysis could be used for accurate determination of fetal sex, we measured testosterone (T) and follicle-stimulating hormone (FSH) in 130 amniotic fluid samples at midgestation. The mean unconjugated T in amniotic fluid of 73 patients carrying male fetuses was 202 pg. per milliliter (95 per cent confidence limits [CL]: 70 to 580) and all but three had levels higher than 90 pg. per milliliter. The mean amniotic fluid unconjugated T for 49 patients carrying female fetuses of 41 pg. per milliliter (95 per cent CL: 11 TO 125) was fivefold lower than that for the male fetuses and all but three patients carrying female fetuses had amniotic fluid T levels of 90 pg. per milliliter or lower. The mean amniotic fluid FSH of 0.7 ml. U. per milliliter (95 per cent CL: less than 0.5 to 3.4) for subjects with male fetuses was tenfold lower than that for patients with female fetuses. Amniotic fluid FSH levels less than 2.0 ml. U. per milliliter were found in 88 per cent of patients carrying male fetuses and in only one patient with a female fetus, and levels greater than 10 ml. U. per milliliter were found in those with female fetuses only. In eight patients (7 per cent of cases), neither amniotic fluid T nor FSH determinations were indicative of fetal sex. Measurement of unconjugated T and FSH in amniotic fluid may be an adjunct to other methods of determining fetal sex.

Amniotic Fluid↗

[Association between amniotic fluid insulin in the second trimester, maternal glucose tolerance and fetal malformations].

INTRODUCTION: Maternal hyperglycemia during gestational diabetes leads to fetal hyperinsulinemia, which is associated with increased perinatal morbidity and mortality. Amniotic fluid insulin levels are therefore considered by some researchers to be ideal parameters to use in diagnosing gestational diabetes and making decisions about correct therapy. There are various recommendations about determining gestational diabetes early in pregnancy (< 24 weeks) by measuring amniotic fluid insulin. This study tests this association -- taking additional risk factors into account -- in a group of pregnant women who had genetic indications requesting for amniocentesis (AC). MATERIALS AND METHODS: All pregnant women who came to our clinic for genetically-determined amniocentesis from April 10, 1995 - Jan. 31, 2000 and who were between 12 and 24 weeks were included in our study. After a sample of amniotic fluid was taken, the laboratory performed a competitive radio-immuno-assay to determine the insulin concentration. O'Sullivan's cut-off values were used in diagnosing gestational diabetes. Since not all pregnant woman in our clinic were screened for gestational diabetes, we gathered our data retrospectively by checking all birth records; these were available in our clinic's data archive. RESULTS: A total of 483 pregnant women were included in our study. 22 (4.6 %) of them were classified as gestational diabetics. The average value for amniotic fluid insulin was 1.21 mU/L +/- 0.89. The insulin values for the entire study population exhibited a weekly increase of 0.1 mU/L from the 12th through the 24th week. The insulin concentrations for the 22 gestational diabetics were not significantly higher than those of the non-diabetics (1.05 mU/L vs. 1.0 mU/L; p = 0.34). In the 90 (th) percentile and above of the amniotic fluid insulin levels (2.2 mU/L) for the entire study population, the rate of gestational diabetics was at 11.8 % three times that of the non-diabetics, at 3.7 % (p = 0.021). Among the risk factors for gestational diabetes, an increased body mass index (BMI) value correlated significantly with increased insulin concentration (p < 0.001). The patients at and above the 90th percentile also had significantly higher BMI values (p = 0.002). In the multivariate analysis, the following influences were determined to be significant: maternal body mass index (p < 0.001) and the gestational age (p < 0,001), not the mere diagnosis of "gestational diabetes". A significant association was not found between elevated insulin values in amniotic fluid and the child's birth weight, APGAR values, pH-levels and blood glucose values. However, a significant association was found regarding fetal malformations and chromosome abnormalities. CONCLUSION: Even very low concentrations of insulin can be identified in amniotic fluid early in the pregnancy. The values increase during the course of the pregnancy. There is a positive correlation between maternal weight (BMI) and insulin levels in the amniotic fluid. Pregnant women with gestational diabetes have higher insulin levels in their amniotic fluid. The multivariant analysis shows, however, that this association can be traced to the maternal BMI and the time point during the pregnancy when the AC was performed. Malformations, especially those with a neural tube defect, are an additional cause for elevated insulin values in amniotic fluid.

Adult↗

The relationship between spontaneous rupture of membranes, labor, and microbial invasion of the amniotic cavity and amniotic fluid concentrations of prostaglandins and thromboxane B2 in term pregnancy.

OBJECTIVE: The purpose of this study was to examine the relationship between rupture of membranes, labor, and microbial invasion of the amniotic cavity and amniotic fluid concentrations of eicosanoids in patients with spontaneous rupture of membranes at term. STUDY DESIGN: Amniotic fluid was retrieved by transabdominal amniocentesis from patients with rupture of membranes and patients with intact membranes at term. Studies to determine the microbial state of the amniotic cavity included culture for bacteria and mycoplasmas, Gram stain, amniotic fluid white blood cell count, and Limulus amebocyte lysate. Eicosanoids (prostaglandin E2, prostaglandin F2 alpha and its stable metabolite, 6-keto-prostaglandin F1 alpha, and thromboxane B2) were determined with sensitive and specific radioimmunoassays validated for human amniotic fluid. Statistical inference was conducted with analysis of variance and linear contrast. RESULTS: (1) Spontaneous rupture of membranes at term was associated with a significant increase in amniotic fluid concentrations of all eicosanoids measured in this study except 6-keto-prostaglandin F1 alpha. (2) Early labor in patients with rupture of membranes was associated with a significant increase in the amniotic fluid concentration of all eicosanoids. (3) A significant increase in amniotic fluid eicosanoids in women with microbial invasion of the amniotic cavity could not be documented. CONCLUSIONS: Whereas preterm labor in the absence of microbial invasion of the amniotic cavity is not associated with a significant increase in amniotic fluid concentrations of prostaglandins, a clear increase was documented in women with early labor after spontaneous rupture of membranes. These observations suggest that there are fundamental differences in the biochemistry of term and preterm parturition.

Amnion↗