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Identification of phospholipid platelet-activating factor (1-0-alkyl-2-acetyl-sn-glycero-3-phosphocholine) in human amniotic fluid and urine.

Human amniotic fluid and fetal urine were examined for the presence of phospholipid platelet-activating factor (PAF). PAF was detected in lipid extracts of some samples of amniotic fluid obtained from women in labor but it was undetectable in samples of amniotic fluid obtained before the onset of labor. PAF was identified by chromatographic mobility, platelet aggregation and chemical modifications. LysoPAF was also present in amniotic fluid at higher concentrations than those of PAF. Both PAF and lysoPAF were identified also in newborn and adult urine.

Amniotic Fluid↗

Ascorbic acid concentration in amniotic fluid in late pregnancy.

Amniotic fluid and venous blood specimens were obtained from 34 pregnant women and analyzed for the ascorbic acid concentration. The mean amniotic ascorbic acid concentration of smoking pregnant women was less than 50% of that of non-smoking women. Pregnant women who smoked had a significantly lower serum and amniotic fluid ascorbic acid concentration than those who did not smoke. No differences were observed between the groups with or without premature rupture of the fetal membrane. The results suggest that ascorbic acid in the amniotic fluid reflects the ascorbic acid status in the blood of pregnant women and smoking had a greater effect in decreasing the ascorbic acid concentration in amniotic fluid than in serum.

Adult↗

[Amnioreduction in treatment of increased amniotic fluid volume].

INTRODUCTION: Increased amniotic fluid volume may significantly increase the risk for preterm delivery. Amniodrainage is a symptomatic treatment by which excess amniotic fluid is reduced to provide fetal lung maturation. The aim of this stuldy is to estimate the efficacy and safety of this procedure, our results and give a literature review. MATERIAL AND METHODS: Sonografic criteria were used (AFI >400 ml, or the biggest amniotic fluid pocket >150 mm) to choose patients in whom 18G needle was used to allow leaking of excessive amniotic fluid. RESULTS: 10 patients underwent 26 procedures. The procedures were performed at 28.6 th week gestation, on average and 6.25 weeks average gain or 1000 g. We had two sets of monochorionic twins with twin-to-twin transfusion syndronme (TTTS), where one child survived. One procedure was followed by premature placental abruption, and premature delivery in 28th week. The rest of procedures were uneventful. CONCLUSION: In our series of 10 women, 26 procedures were performed to prolong pregnancies, enable fetal maturation and weight gain. In majority of cases amnioreduction was done without complications, so we could repeat the intervention and prolong the pregnancy. Survival of one child in two TTTS pregnancies should not be regarded unsulccessful in our conditions.

Amniotic Fluid↗

Monocyte chemotactic protein-1 in cervical and amniotic fluid: relationship to microbial invasion of the amniotic cavity, intra-amniotic inflammation, and preterm delivery.

OBJECTIVE: The purpose of this study was to evaluate the role of monocyte chemotactic protein-1 in cervical and amniotic fluid in women in preterm labor and with preterm premature rupture of membranes. STUDY DESIGN: Women with singleton pregnancies (<or=34 weeks) in preterm labor (n=75 women), with preterm premature rupture of membranes (n=47 women), and at term (n=45 women) who were undergoing elective cesarean delivery were included. Cervical and amniotic fluid were sampled. RESULTS: Monocyte chemotactic protein-1 in cervical and amniotic fluid was higher in women in preterm labor than in women at term. Cervical monocyte chemotactic protein-1 in women in preterm labor was associated with microbial invasion of the amniotic cavity, intra-amniotic inflammation, delivery within 7 days, and at <or=34 weeks. Amniotic monocyte chemotactic protein-1 correlated to microbial invasion of the amniotic cavity in women with preterm premature rupture of membranes, intra-amniotic inflammation in preterm labor, preterm premature rupture of membranes, delivery within 7 days, and delivery at <or=34 weeks in women in preterm labor. CONCLUSION: Monocyte chemotactic protein-1 in cervical and amniotic fluid levels are elevated in preterm labor and preterm premature rupture of membranes and correlate to intra-amniotic infection/inflammation.

Amniotic Fluid↗

Raised hypoxanthine, xanthine and uridine concentrations in meconium stained amniotic fluid and during labour.

Amniotic fluid samples were obtained at induction of labour in 64 women; in 15 of these there was meconium staining of the amniotic fluid; the remainder showed no signs of fetal distress. Using high pressure liquid chromatography, compared to the samples from normal patients there were highly significantly raised levels of hypoxanthine, xanthine and uridine in the meconium stained samples; oxypurines in the meconium itself could not explain the difference. Where serial samples were obtained during labour by intrauterine catheter, a terminal rise in oxypurine levels was apparent. Where the proportion of oxypurine present as hypoxanthine exceeded one per cent in amniotic fluid at the time of induction, there was a significantly greater occurrence of late fetal heart rate decelerations in the ensuing labour. These findings are consistent with other evidence that when tissues become hypoxic the metabolic products of nucleotide breakdown escape from the cells and appear in extracellular fluid. Oxygen lack in the fetus probably causes loss of these compounds from the hypoxic kidneys to the urine so that they appear in amniotic fluid.

Amniotic Fluid↗

Antimicrobial effect of amniotic fluid against anaerobic bacteria.

Amniotic fluid samples were obtained at term and tested for their antimicrobial effect on anaerobes, Peptostreptococcus (Ps.) anaerobius, Peptococcus (Pc.) prevotii, Bacteroides (B.) fragilis, and B. coagulans with facultative Escherichia (E.) coli serving as control. Amniotic fluid had only temporary bacteriostatic effect on Pc. prevotii and B. fragilis for 8 to 16 hours. This effect lasted for only 8 hours on Ps. anaerobius. On the contrary, the bacteriostatic effect of amniotic fluid was well sustained on E. coli and B. coagulans, lasting for the entire test periods of 20 and 32 hours, respectively. At the end of the time intervals mentioned, Ps. anaerobius, Pc. prevotii, and B. fragilis exhibited logarithmic growth, confirming the earlier reports that it is not nutritionally deficient. Amniotic fluid exhibited temporary bacteriostatic effect on Ps. anaerobius, P. prevotii, and B. fragilis, but this effect was well sustained against B. coagulans.

Amniotic Fluid↗

Analysis for amniotic fluid crystallization in second-trimester amniocentesis.

A potential complication of second-trimester amniocentesis for genetic indications is inadvertent needle insertion into the maternal bladder, resulting in aspiration of urine rather than amniotic fluid. Amniotic fluid forms a characteristic crystalline arborization pattern when allowed to air dry. We utilized this property of amniotic fluid to distinguish amniotic fluid from maternal urine. In 24 of 25 cases studied in a randomized blind fashion the crystalline arborization test correctly identified amniotic fluid, whereas none of the 25 urine samples showed this pattern. Our study indicates that the crystalline arborization test is reliable in distinguishing amniotic fluid from maternal urine during the second trimester of pregnancy.

Amniocentesis↗

Meconium-stained amniotic fluid: a risk factor for microbial invasion of the amniotic cavity.

The purpose of this study was to determine whether meconium-stained amniotic fluid is a marker for microbial invasion of the amniotic cavity. Amniocentesis was performed on 707 patients presenting with preterm labor and intact membranes. Meconium-stained amniotic fluid was present in 4.2% (30/707) of patients with preterm labor. The prevalence of positive amniotic fluid cultures was significantly higher in women with meconium-stained amniotic fluid than in women with clear fluid (33% [10/30] vs 11% [75/677]; p = 0.001; odds ratio = 4.01; 95% confidence interval = 1.6 to 9.4). Patients with meconium-stained amniotic fluid were also more likely to have failed tocolysis and delivered a preterm neonate more frequently than patients with clear fluid (83% [25/30] vs 38% (258/677); p = 0.0001; odds ratio = 8.1; 95% confidence interval = 2.9 to 24.4). We conclude that meconium-stained amniotic fluid is a risk factor for microbial invasion of the amniotic cavity and preterm delivery in women with preterm labor and intact membranes.

Amniotic Fluid↗

Maternal smoking induced cotinine levels and genotoxicity in second trimester amniotic fluid.

Cotinine concentrations in amniotic fluid samples from 22 smoking and 37 non-smoking pregnant women and induction of sister-chromatid exchanges (SCE) in Chinese hamster ovary (CHO) cells by samples from 15 smokers and 15 non-smokers were studied as indicators of exposure to potential genotoxic activity during pregnancy. Analysis of cotinine revealed one individual in the non-smoking group with a high cotinine level apparently due to non-reported smoking. The mean cotinine concentration of smokers was 85 ng/ml whereas non-smokers had a concentration of 0.3 ng/ml. According to interview data 16 persons announced some passive exposure to tobacco smoke at home or at work; however this group did not differ from unexposed non-smokers in their amniotic fluid cotinine concentration. SCE inducing activity was tested with and without metabolic activation. The mean SCE frequency in CHO cells induced in the presence of exogenous metabolic activation by concentrated amniotic fluid of heavy smokers (> or = 10 cigarettes/day) was significantly higher (9.7 +/- 0.6 SCE/cell) than among non-smokers (8.9 +/- 0.6 SCE/cell) with metabolic activation. The results show that amniotic fluid cotinine measurements and induction of SCEs in CHO cells can be used to indicate fetal exposure by maternal smoking and support earlier studies suggesting a potential genotoxic hazard to the fetus of heavy smokers.

Adult↗

Relationship of amniotic fluid markers of intra-amniotic infection with histopathology in cases of preterm labor with intact membranes.

OBJECTIVE: To evaluate the correlation of amniotic fluid (AF) markers (AFMs) of intra-amniotic infection with histopathologic findings in cases of preterm labor with intact membranes, between 22 and 36 weeks' gestation. STUDY DESIGN: We reviewed the charts of patients admitted in preterm labor with intact membranes between January 1993 and December 1996. Those having amniocentesis were identified, and AFMs were compared with histopathology in patients who delivered within 48 hours of the amniocentesis. The AFMs evaluated were glucose, polymorphonuclear leukocytes, Gram stain, and culture. All placentae were reviewed by a single pathologist blinded to the AF findings. Histologic evidence of acute inflammation was defined by findings of both subchorial intervillositis and marginating choriodeciduitis. The sensitivities, specificities, and positive and negative predictive values of the various AFMs were calculated. RESULTS: Of 556 women with intact membranes presenting in preterm labor, 181 (32.6%) had amniocentesis and 88 delivered within 48 hours of the amniocentesis. Histopathologic chorioamnionitis was seen in 53 patients (60.2%). The findings (with their sensitivity, specificity, and positive and negative predictive values) were: polymorphonuclear leukocytes at > 10/high-power field (22.6%, 97.2%, 92.3%, and 46.1%), positive Gram stain (26.4%, 94.6%, 87.5%, and 47.3%), culture (28.3%, 92.1%, 83.3%, and 47.9%), and glucose of < 15 mg/dl (28.3%, 94.6%, 88.2%, and 47.9%), respectively. Using a receiver-operator characteristic curve for different level of AF glucose, a glucose level of < 20 mg/dl was the most sensitive AF predictor of histologic chorioamnionitis. CONCLUSION: Histopathologic evidence of chorioamnionitis was present in 60.2% of cases of preterm births due to preterm labor in women who at our institution were offered and accepted amniocentesis and subsequently delivered within 48 hours. AFMs may be useful predictors of histologic chorioamnionitis. The most efficient AFM for chorioamnionitis in this group of patients was glucose at < 20 mg/dl.

Adult↗

Enzymatic measurement of phosphatidylglycerol in amniotic fluid.

We describe an enzymatic method for measuring phosphatidylglycerol (PG) at concentrations as low as 0.2 mumol/L in amniotic fluid. Amniotic fluid (1.5 mL) is centrifuged at 10,000g for 20 min to obtain a lamellar body pellet, which is reconstituted with 0.5 mL of buffer. The PG is measured by a two-step enzymatic scheme. Recovery studies demonstrated that the pellet contains > 97% of the PG present in amniotic fluid. Between-run CVs were 28%, 5.7%, and 2.6% for amniotic fluid controls with means of 0.32, 3.9, and 10.7 mumol/L, respectively (n = 20). The enzymatic procedure was not significantly affected by blood, meconium, bilirubin, or other phospholipids. Lecithin/sphingomyelin ratio (n = 101) and fluorescence polarization (n = 127) compared with log(PG) showed correlation coefficients of 0.832 and -0.866, respectively. This test's ability to detect low concentrations of PG in amniotic fluid may make it a better predictor of fetal lung immaturity than previous methods.

Amniotic Fluid↗

Evidence for somatostatin precursors in human stomach, placenta, and amniotic fluid.

Human stomach, placenta, and amniotic fluid have previously been shown to contain immunoreactive somatostatin (IRS). The present studies were undertaken to further characterize this IRS. Gel chromatography of amniotic fluid revealed only one peak of somatostatin-like immunoreactivity (SLI; mol wt, 15,000) regardless of gestational age. Extracts of human fetal stomach contained three peaks of SLI: 87% of the total IRS coeluted with synthetic tetradecapeptide somatostatin (SRIF), 12% coeluted with synthetic somatostatin-28 (S-28), and 4% coeluted with amniotic fluid SLI. Extracts of 9- to 13-week-old placentas contained 38.9 +/- 5.3 pg IRS/mg protein (range, 21-62 pg IRS/mg protein). Chromatography revealed that 57% of the total IRS coeluted with SRIF, 19% coeluted with S-28, and 23% eluted in a position indicating a molecular weight of 12,000. Serial dilutions of amniotic fluid SLI and material from each peak of stomach and placental SLI showed parallelism with synthetic SRIF. Treatment with 8 M urea and dithiothreitol did not convert any of these SLIs to smaller immunoreactive forms. Incubation of purified amniotic fluid SLI with 1% (wt/wt) L-(tosylamido 2-phenyl)ethyl chloromethyl ketone-trypsin for 90 min resulted in partial conversion to immunoreactive material coeluting with SRIF. When synthetic S-28 was incubated in fresh amniotic fluid at 37 degrees C, it was rapidly degraded (t 1/2 approximately or equal to 25 min). These studies indicate that human amniotic fluid IRS is composed of 15K SLI only, whereas human stomach and placental IRS are heterogeneous, comprising SRIF as well as larger forms of SLI which probably represent SRIF precursors.

Amniotic Fluid↗

Amniotic fluid soluble fas levels in intra-amniotic infection.

OBJECTIVE: Membrane Fas can induce apoptosis in sensitive cells. It has been reported that soluble Fas (sFas) is elevated in septicemia. We examined amniotic fluid (AF) sFas levels in patients with and without intra-amniotic infection. METHODS: Forty-two AF specimens were studied. Intra-amniotic infection was defined as the presence of a positive AF culture. Twenty-one specimens were from patients with intra-amniotic infection and 21 were from patients without intra-ammotic infection. Amniotic fluid sFas was determined by an enzyme immunoassay and normalized by AF creatinine levels. The Mann-Whitney U test, contingency table method, and Spearman's rank correlation test were used for statistical analyses. Data were expressed as median with ranges. RESULTS: There were no significant differences in maternal age, gestational age, parity, and race between the groups. The median AF sFas was significantly higher with intra-amniotic infection than without it (5.07 U/mL, range 0.32-13. 25 compared with 1.95 U/mL, range 0.01-5.35; P =.004). After normalizing to AF creatinine, infected fluids also had significantly higher median sFas/creatinine than uninfected amniotic fluids (289.1 U/mg creatinine, range 16.6-920.5 compared with 126.8 U/mg creatinine, range 0.5-546.2; P =.01). Amniotic fluid sFas and sFas/creatinine were positively correlated with AF leukocytes and negatively correlated with AF glucose. CONCLUSION: Elevated AF sFas is associated with intra-amniotic infection. High production of AF sFas in intra-amniotic infection may play a role in the inhibition of apoptosis of AF leukocytes, leading to the persistence of inflammation.

Adolescent↗

[Cervix reconstruction after amniotic fluid puncture in amniotic sac prolapse in the second trimester].

The possibility and the problems of an surgical technique because of cervical insufficiency in the 2. trimester with a hourglass amniotic prolapse is shown in three cases. A new concept including amniocentesis and doubleclerclage is developed with a wide physiological reconstruction of the cervix and its occlusion mechanism at the inner and outer cervix. We show an alternative to previous surgical techniques that makes a prolongation of the pregnancy possible and causes additionally because of the minimal surgical procedure no disadvantage for the cervical function in coming pregnancies. Small surgical strain, quick surgical procedure and simultaneously continue of diagnostic possibilities as well as the reconstructive technique with small trauma of the cervical tissue are the advantages of this method.

Adult↗

Do semiquantitative amniotic fluid indexes reflect actual volume?

OBJECTIVE: Our objective was to determine how well semiquantitative ultrasonographic measures of amniotic fluid, i.e., maximal amniotic fluid vertical pocket and amniotic fluid index, reflect actual amniotic fluid volumes in 50 near-term patients whose amniotic fluid volume estimates were normal by visual inspection. STUDY DESIGN: Before amniocentesis for fetal lung maturity, each patient had visual amniotic fluid volume estimates, maximal amniotic fluid vertical pocket, and amniotic fluid index performed by the same examiner, and then each received intraamniotic injection of a 10% paraaminohippurate solution. Amniotic fluid volume was quantitated by spectrophotometric assay of paraaminohippurate concentration. Oligohydramnios and polyhydramnios were defined as < 300 and > 2000 ml, respectively. RESULTS: Quantitative amniotic fluid volume was positively related to both amniotic fluid index and maximal amniotic fluid vertical pocket (r = 0.75 and 0.60, respectively). True-positive rates for oligohydramnios (amniotic fluid index < 5 cm or maximal amniotic fluid vertical pocket < 2 cm) were 100% and 0%, respectively; false-positive rates with either method were 0%. True-positive rates for polyhydramnios (amniotic fluid index > 20 cm and maximal amniotic fluid vertical pocket > 8 cm) were 0%; false-positive rates were 16% and 24%, respectively. CONCLUSIONS: Amniotic fluid index appears to be slightly better than maximal amniotic fluid vertical pocket for reflecting actual amniotic fluid volume. Both indirect methods tend to overestimate actual amniotic fluid volume at the upper end of its extremes.

Amniotic Fluid↗

Rapid determination of fetal sex in coelomic and amniotic fluid by fluorescence in situ hybridisation.

Coelomic fluid (n = 32), amniotic fluid (n = 26) and placental tissue were obtained from 32 women undergoing termination of pregnancy at 7-11 weeks of gestation. Fluorescence in situ hybridisation (FISH) was performed to determine fetal sex using a heterochromatic Y probe and an alpha satellite repeat probe for chromosome X. In each case there was concordance in the fetal sex from the three compartments. However, no result could be obtained from 4 of the 32 coelomic fluids and 4 of the 26 amniotic fluid samples. The hybridisation efficiency was significantly lower in coelomic and amniotic fluid compared with placental tissue.

Amniotic Fluid↗