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Erythropoietin levels in amniotic fluid and extraembryonic coelomic fluid in the first trimester of pregnancy.

OBJECTIVE: The aim was to measure erythropoietin levels in amniotic fluid and extraembryonic coelomic fluid from 7-12 weeks' gestation. SUBJECTS: Twenty healthy women with ultrasonographically normal first trimester pregnancies prior to surgical termination. METHODS: Paired samples of amniotic fluid and extraembryonic coelomic fluid were collected by transvaginal ultrasound guided needling. Erythropoietin was measured in both pregnancy fluids using a radioimmunoassay. RESULTS: There was a highly significant difference between erythropoietin levels in extraembryonic coelomic fluid (median level 15.45 mU/ml; range 6.8-32.1 mU/ml) and those in amniotic fluid (median 5.0 mU/ml; range < 5.0-5.8 mU/ml) (P < 0.0001; Mann-Whitney U-test). The levels of erythropoietin in maternal serum (median 15.4 mU/ml; range 5.6-29.4 mU/ml) were similar to those in the extra-embryonic coelom (P = 0.81; Mann-Whitney U-test). No relation was demonstrated between erythropoietin levels in amniotic fluid or coelomic fluid and stage of gestation. CONCLUSION: High levels of erythropoietin in coelomic fluid suggests that the hormone is involved in the process of human extraembryonic erythropoiesis. The exact regulatory role remains unknown.

Amniotic Fluid↗

Amniotic fluid tumor necrosis factor-alpha and the risk of respiratory distress syndrome among preterm infants.

OBJECTIVE: We examined the effect of exposure to amniotic fluid infection and cytokines on the pulmonary outcome of preterm infants. STUDY DESIGN: A cohort of 136 preterm infants born to women in preterm labor had amniotic fluid cultures and tumor necrosis factor-alpha assays performed. Amniotic fluid was collected by transabdominal amniocentesis. Outcome measures included respiratory distress syndrome and length of oxygen and ventilator support. Logistic regression and Cox proportional hazards regression adjusted for birth weight and other confounders. RESULTS: Respiratory distress syndrome developed in 67 (49%) of 136 infants. Elevated amniotic fluid tumor necrosis factor-alpha levels and amniotic fluid infection were significantly associated with respiratory distress syndrome. This association persisted after adjustment for birth weight. Infants exposed to tumor necrosis factor-alpha remained on supplemental oxygen and assisted ventilation longer and had longer hospital stays compared with nonexposed infants. CONCLUSION: Prenatal exposure to tumor necrosis factor-alpha may be a risk factor for respiratory distress syndrome and its complications.

Amniocentesis↗

Amniotic fluid OD 650 as a rapid indicator of fetal pulmonary maturity.

Amniotic fluid OD 650 (optical density at 650 nm) greater than 0.15 has been associated with mature L/S ratios. This study compared OD 650 with the L/S ratio on 235 uncontaminated amniotic fluid specimens. Follow-up was obtained on 138 neonates delivered within 72 hours of amniocentesis. Of 124 amniotic fluid specimens with OD 650 greater than 0.15, 109 had mature L/S ratios. Only 3 of 96 neonates with OD 650 greater than 0.15 developed respiratory distress syndrome; 8 of 42 neonates with OD 650 values less than 0.15 developed it. The false-positive rate (2.1%) compared favorably with that of the L/S ratio (2.9%). A higher false-negative rate (24.6%) decreased the efficiency of OD 650 as an indicator of fetal pulmonary maturity. An OD 650 greater than 0.15 appears to be a rapid, reliable indicator of fetal lung maturity.

Amniotic Fluid↗

Mechanisms involved in the stimulatory effect of amniotic fluid on prostaglandin production by human fetal membranes.

This study aims to investigate potential mechanisms involved in the stimulatory effect of amniotic fluid on prostaglandin production by fetal membranes. A cell culture study of amnion and chorion was obtained following elective caesarean section, incubated with amniotic fluid collected at term (37-42 weeks' gestation) following either spontaneous labour (n = 6) or elective caesarean section (n = 6). The effect of addition of cycloheximide and actinomycin D (inhibitors of translation and transcription respectively), and staurosporine and genistein (inhibitors of protein kinase C and tyrosine kinase respectively) to these cultures was investigated. ANOVA was employed for statistical analysis. Cycloheximide and staurosporine significantly inhibited the stimulatory effect of spontaneous labour and elective section amniotic fluid on PGE2 production by amnion, and PGEM production by chorion. Genistein significantly inhibited the stimulatory effect of spontaneous labour amniotic fluid on PGE2 and PGEM production by amnion and chorion respectively. The stimulatory effect of amniotic fluid on prostaglandin production is dependent on new protein synthesis, presumably cyclooxygenase (COX), and stimulation of cell signal transduction pathways involving protein kinase C and tyrosine kinase.

Amniotic Fluid↗

Demonstration of haemagglutinating antibody against food antigens in the amniotic fluid and cord blood.

1) Haemagglutinating antibody against egg white was demonstrated with higher incidence and with higher titres in the amniotic fluid than in sera obtained from the mother, cord blood, and newborn. On the other hand antibodies were barely detectable or absent from the meconium newborn infant's first voided urine. 2) In the amniotic fluid, haemagglutinating antibody was detected in highest titres against soybean and lactalbumin, respectively. 3) There was a similar trend of distribution of haemagglutinating antibody against egg white, lactalbumin or soybean in amniotic fluids from the same individual. 4) Haemagglutinating antibody against egg white in the amniotic fluid was identified as an immunoglobulin belonging to the IgM class; the possibility of the fetal origin of the haemagglutinating antibody in amniotic fluid was discussed.

Agglutinins↗

Amniotic fluid alpha-fetoprotein is not a useful biological marker of pregnancy outcome.

The aim of our study was to determine if the amniotic fluid alpha-fetoprotein (AFP) level could be a useful predictive biochemical marker of pregnancy outcome. Amniotic fluid AFP measurement was prospectively carried out over a three-year period. After excluding factors susceptible to modifying AFP measurements, 587 subjects with gestational age between 14 and 20 weeks were selected to compare the amniotic fluid AFP mean levels depending on the occurrence of an adverse outcome. No significant associations between amniotic fluid AFP level and poor pregnancy outcome, i.e. pre-eclampsia, preterm delivery, premature rupture of fetal membranes, fetal growth retardation and placental abnormalities were observed. The routine measurement of amniotic fluid alpha-fetoprotein during an amniocentesis procedure was not considered useful in predicting pregnancy complications.

Adult↗

Fluid transport across human fetal membranes affected by human amniotic fluid prolactin: an in vitro study.

The aim of the study was to investigate the effects of human amniotic fluid (AF) prolactin (PRL) on the transfer of fluids across human fetal membranes, detected in vitro by weight change. We have developed a system, consisting of an inner compartment (4 ml), suspended in an outer compartment (40 ml), and separated from it by a 2 cm2 circular piece of fresh human fetal membrane (amnion, chorion and adhering decidua). The weight of the inner compartment was continuously registered with an electronic precision balance. Osmolality changes or hydrostatic pressure differences did not affect the rate of mass transfer through the membranes. Potassium cyanide significantly influenced the mass transfer in favour of the applied hydrostatic pressure difference (+2 cm H2O to the amniotic side) or osmotic pressure difference (amniotic side 265 mosm/kg/decidual side 285 mosm/kg). The PRL preparations used were human and ovine pituitary PRL, as well as three fractions isolated from human AF by fractionated ammonium sulphate and ethanol precipitations, followed by Sephacryl chromatography. All PRL preparations were tested in physiologic concentrations (0.5 to 5 micrograms/ml). Only the two largest AF-PRL variants significantly disturbed the balance of fluid transfer across the membranes when added to the fetal (amniotic) side of the membrane. This resulted in a net increase in fetomaternal transfer of 120 to 180 microliters. This effect could be repeated and lasted for at least 25 minutes. Using an antibody against hPRL the effect was completely blocked. Neither AF-PRL added to the maternal (decidual) side of the membrane nor oPRL or human pituitary PRL added to the amniotic or decidual side changed the rate of mass transfer across the membranes.

Amniotic Fluid↗

Fetal lung maturatio. III. Amniotic fluid phosphatidic acid phosphohydrolase (PAPase) and its relation to the lecithin/sphingomyelin ratio.

The purpose of this study was to establish 1) the sequential changes in specific activity of phosphatidic acid phosphohydrolase (PAPase) in amniotic fluid and its relation to the lecithin/sphingomyelin (L/S) ratio; and 2) the origin of amniotic fluid PAPase. the increase in the amniotic fluid L/S ratio is preceded by an increase in PAPase activity, rising from 15 nmoles of phosphate released per milligram of protein per hour at 30 weeks to 100 nmoles at 37 weeks. The mean PAPase activity in the nasopharyngeal fluid of the infant is 456 nmoles of phosphate released per ml per hour, the amniotic fluid mean PAPase activity at delivery being 129 nmoles (P less than 0.01). These findings are consistent with the view that amniotic fluid PAPase originates, in part, from the fetal lung and likely participates in the regulation of the synthesis of lecithin.

Amniotic Fluid↗

Assessment of fetal pulmonic maturity by measurement of the surface tension of amniotic fluid lipid extract.

Fetal pulmonic maturity is currently predicted by the lecithin/sphingomyelin (L/S) ratio which utilizes the changes with increased gestation in the lecithin component of amniotic fluid surfactant. The surface tension lowering property of lipids extracted from the amniotic fluid of 45 patients was measured directly. The surface tension (gamma) of various aliquots of the amniotic fluid lipid extract was measured by a modification of the Wilhelmy plate method. At 80 gamma of the extract, a gamma of 67 dynes/cm or less indicated pulmonic maturity, while a gamma of greater than 67 dynes/cm suggested immaturity, provided that the gamma at 200 gamma was greater than 56 dynes/cm. The fluid was termed transitional if the gamma was greater than 67 dynes/cm at 80 gama, but less than 56 dynes/cm at 200 gamma. A comparison was made with the L/S ratio; there were no false positive values and only 13.6% false negative surface tension values. Addition of blood to amniotic fluid, mature and immature, yielded the same results as with the L/S ratio. Two infants with respiratory distress syndrome had both immature L/S ratios and surface tension results. Results indicate that the surface tension of the amniotic fluid lipid extracts is a reliable and rapid means of antenatally predicting fetal pulmonic maturity.

Amniotic Fluid↗

Developmental patterns of intestinal disaccharidases in human amniotic fluid.

A close relationship exists between relative disaccharidase activities (maltase, sucrase, trehalase, palatinase, turanase, lactase, and cellobiase) in amniotic fluid and corresponding jejunal mucosa of five human fetuses (16 to 21 weeks of gestation) suggesting that these intestinal enzymes pass into amniotic fluid. Serial determination of disaccharidase activities in amniotic fluid samples collected between 10 and 42 weeks of gestation showed maximum mean activities at 14 to 17 weeks of gestation and a rapid drop to less than 12 per cent maximum values at about 22 weeks. This drop is probably caused by combined effects of decreased extrusion rate of intestinal disaccharidases and increased reabsorption of the enzymes in swallowed amniotic fluid with fetal development.

Amniotic Fluid↗

Atrial natriuretic factor in amniotic fluid and in maternal venous blood of pregnancies with fetal cardiac malformations and chromosomal abnormalities.

OBJECTIVE: We evaluated the levels of atrial natriuretic factor (ANF) in amniotic fluid and in maternal venous blood in pregnancies with fetal cardiac malformations and chromosomal abnormalities. METHOD: Between the 16th and 18th week of pregnancy, 151 women were divided into three groups. Group A included patients at lowest risk, carrying a fetus with a normally developing heart and normal karyotype (control group). Group B included women with a fetus suffering from cardiac malformations, with or without associated chromosomal abnormalities. Group C included women carrying a fetus affected with chromosomal abnormalities without congenital cardiopathies. ANF was evaluated by radioimmunoassay. RESULTS: In maternal venous blood, the mean levels of ANF were 42.1, 53.1 and 38.7 pg/ml in groups A, B and C, respectively. In amniotic fluid, the mean levels of ANF were 34.2, 101.8 and 35.8 pg/ml in groups A, B and C, respectively. In group A (control group) there was no statistical difference in ANF levels across the gestational age range of 16-18 weeks, either in amniotic fluid or in maternal venous blood. A significant difference of ANF content in maternal venous blood was revealed in comparing group A with group B (p < 0.01), and group C with group B (p < 0.01). A statistically significant difference in ANF levels was also found in amniotic fluid between group A and group B (p < 0.01), and between group C and group B (p < 0.01). No statistically significant differences were found between group C and group A in comparing ANF levels in maternal venous blood and amniotic fluid. CONCLUSION: ANF levels in amniotic fluid and in maternal venous blood are increased early in the case of fetuses with cardiac malformations, with or without associated karyotype alteration. Chromosomally abnormal fetuses without heart malformations have normal ANF levels. These results could be useful for elucidating fetal pathophysiology mechanisms.

Adult↗

The relationship between acute inflammatory lesions of the preterm placenta and amniotic fluid microbiology.

OBJECTIVE: Our objective was to determine the relationship between microbial invasion of the amniotic cavity and the presence and severity of acute inflammatory lesions in the placenta. STUDY DESIGN: Placental histologic and amniotic fluid microbiologic studies were performed in 92 consecutive patients who were admitted with preterm labor and intact membranes and delivered within 48 hours after amniocentesis. RESULTS: The prevalence of a positive amniotic fluid culture was 38% (35 of 92). There was a strong association between the presence and severity of inflammation in the amnion, chorion-decidua, umbilical cord, and chorionic plate and the results of the amniotic fluid culture (p less than 0.0001 for each tissue section). Three patterns of inflammation of the chorion-decidua were identified: marginating, nonmarginating, and a mixed pattern. The marginating and the mixed patterns of inflammation were strongly associated with the presence of a positive amniotic fluid culture. Acute inflammation of the chorionic plate was the most sensitive indicator of a microbial invasion of the amniotic cavity (sensitivity 96.6%), and funisitis and umbilical vasculitis had the highest specificity (85.7%). CONCLUSION: The presence of acute inflammatory lesions of the chorioamniotic membranes can serve as a marker of microbial invasion of the amniotic cavity.

Amnion↗

Effect of human and recombinant IFN-alpha and IFN-beta on the sister-chromatid exchange (SCE) frequency in amniotic fluid cells in vitro.

Sister-chromatid exchange (SCE) and rates of proliferation in human amniotic fluid cells from healthy donors exposed to human IFN-alpha and IFN-beta and recombinant IFN-alpha and -beta were investigated. Amniotic fluid cells were obtained from pregnant women undergoing genetic amniocentesis. For 46 h, cells were treated with IFNs at concentrations of 10(3)-10(5) U/l. A dose-depending decrease of SCE rate with IFN-alpha and IFN-beta was observed. Our studies in amniotic fluid cells show that the mean SCE frequencies are reduced after incubation with IFN-alpha as well as with IFN-alpha. In contrast to IFN-gamma, the type I IFNs IFN-alpha and IFN-beta cause a genetic effect on DNA repair or a protection from DNA damage. Previously we had shown that a significant dose-depending increase of SCE rates was found in amniotic fluid cultures after addition of IFN-gamma. Therefore, IFN-alpha and IFN-beta (both human IFNs) and also recombinant IFN-alpha and IFN-beta, also in high doses, are neither genotoxic/clastogenic nor embryotoxic. Amniotic cells are vulnerable human cells, which may be well suited for examining the effects of agents like interferon.

Amniotic Fluid↗

Cortisol levels in amniotic fluid in prostaglandin F2alpha-induced midtrimester abortion.

Cortisol concentrations in amniotic fluid and maternal plasma were determined by a competitive protein-binding assay in 14 patients in the midtrimester of pregnancy before and after intra-amniotic administration of prostaglandin F2alpha. Samples were obtained at three-hour intervals until abortion or until the fetal heartbeat ceased. The mean plasma levels of cortisol increased from 324 +/- 173 ng. per milliliter at hour 0 to 524 +/- 272 ng. per milliliter at 6 hours (the peak elevated value). In amniotic fluid, the mean levels of cortisol increased from 4.71 +/- 2.5 ng. per milliliter to a maximum at 9 hours of 10.24 +/- 2.5 ng. per milliliter. We concluded that at 16 to 20 weeks' gestation the fetoplacental unit is capable of responding to prostaglandin F2alpha instillation by increased levels of cortisol in the amniotic fluid.

Abortion, Induced↗

Are amniotic fluid C-reactive protein and glucose levels, and white blood cell counts at the time of genetic amniocentesis related with preterm delivery?

OBJECTIVE: To compare women with spontaneous preterm delivery before 37 weeks and women who delivered at term with respect to amniotic fluid C-reactive protein (CRP), glucose levels, and white blood cell counts at the time of genetic amniocentesis. STUDY DESIGN: The study was conducted on 216 pregnant women who underwent genetic amniocentesis between the 15th and 18th weeks of gestation at Baskent University Obstetrics and Gynecology Department. All patients were followed until delivery for the occurrence of pregnancy complication. Indications for amniocentesis included abnormal triple test results showing increased risk for Down's syndrome, advanced maternal age and sonographic findings indicative for chromosomal abnormalities. The samples were carried immediately to the laboratory for cytogenetic and biochemical examination. Women with spontaneous preterm delivery before 37 weeks (n = 20) and those who delivered at term (n = 196) were compared with respect to some maternal and infant characteristics, amniotic fluid C-reactive protein, glucose levels, and amniotic fluid white blood cell counts. RESULTS: During the study period 244 patients underwent amniocentesis. A chromosomal abnormality was present in 11 patients. 1 patient had a spontaneous pregnancy loss within 3 weeks after the procedure and 16 patients were delivered for fetal or maternal indications (preeclampsia, fetal growth restriction, placenta previa). The remaining 216 women were included in the study and investigated for the risk of preterm delivery. The prevalence of spontaneous preterm delivery before 37 weeks was 9.3% (20/216). There were no significant differences between the preterm delivery and the term delivery groups with respect to C-reactive protein levels and white blood cell counts. Mean amniotic glucose levels were significantly lower in the preterm delivery group (P<0.05). Amniotic fluid glucose levels of < or = 46 mg/dL had a sensitivity of 100% and NPV of 100%. CONCLUSION: Amniotic fluid glucose levels at the time of genetic amniocentesis are lower in women with spontaneous preterm delivery before 37 weeks compared to those who delivered at term. Amniotic fluid glucose levels of < or = 46 mg/dL at the time of genetic amniocentesis may be more sensitive, cheaper and have higher negative predictive value than C-reactive protein levels and white blood cell counts for the prediction of patients in spontaneous preterm labor. The greatest benefit of amniotic fluid glucose testing might be when the physician judges the patient to be at low risk for preterm delivery.

Amniocentesis↗

Detection of hepatitis C virus RNA (HCV RNA) in amniotic fluid: a prospective study.

BACKGROUND/AIMS: Mother-to-infant transmission of hepatitis C virus (HCV) has been reported, but the transmission route is unknown. The aim of our study was to detect HCV RNA in amniotic fluid of pregnant women seropositive for HCV. METHODS: Twenty-two HCV seropositive women were included in the study (median age: 39 years). An amniocentesis was performed in all patients during the 4th month of pregnancy. Sixteen women also tested positive for HCV RNA in serum. The range of HCV RNA titers was 0.3 to 15.1x10(6) Eq/ml (Quantiplex HCV RNA 2.0 Assay, Chiron Diagnostics). Of these 16 viremic patients, four had an anterior placenta, ten had a posterior placenta and the position of the placenta was not determined in two cases. PCR (Amplicor HCV, Roche Diagnostics) was used to detect HCV RNA in the amniotic fluid. We also studied 11 HCV seronegative women as a control group. RESULTS: In the viremic group (n = 16), HCV RNA was detected once in amniotic fluid. The positive specimen was collected from a patient with an HCV RNA serum value equal to 1.1x10(6) Eq/ml. The placenta was in an anterior position. A PCR inhibitor was detected in one case. No HCV RNA was detected in the amniotic fluid of six seropositive non-viremic patients, nor in the control group. Serum HCV RNA was negative in the ten children tested. The woman whose amniotic fluid contained HCV RNA was the mother of one of them. CONCLUSIONS: HCV RNA detection in amniotic fluid is rarely positive. The anterior position of the placenta in the only positive detection cannot rule out contamination of the amniotic fluid during the transplacental amniocentesis.

Adult↗

Amniotic fluid phospholipid analysis in normal and complicated pregnancies.

Amniotic fluid lecithin/sphingomyelin (L/S) ratios were determined at least once in 190 pregnancies. In 127 pregnancies an amniotic fluid specimen was obtained within 72 hours of delivery. This constitutes the corrected group. Respiratory distress was encountered 21 times with 13 of these being clinical hyaline membrane disease (CHMD). The remaining cases were either transient respiratory distress or felt to be aspiration pneumonia. When the L/S ratio was positive and the infant delivered vaginally, there was only one case of CHMD. However, when the patients had grave enough disease to warrant cesarean section, CHMD was encountered in seven cases out of 68 cesarean sections. Of the eight cases of CHMD, five were in mothers whose pregnancy was complicated by diabetes mellitus.

Amniocentesis↗

Volume and glucose concentration of rat amniotic fluid: effects on embryo nutrition and axis rotation.

In order to pursue our previous studies of the changes in neural tube microvilli produced by glucose, we developed a micro method of measuring glucose concentration in the very small volumes of amniotic fluid during neurulation. The volume of amniotic fluid was found to increase nearly 10-fold during major neurulation (day 10 to day 11 in the rat). This increase in volume and our repeated observations that physical removal of the restraining amnion initiates embryonic rotation leads us to propose that the growth of the amniotic cavity is essential for conversion from the ventral- to dorsi-flexion of the embryonic axis. Amniotic fluid volume continues to increase until day 18 but dropped by day 20. A method for glucose determination was developed using the color reaction on glucose oxidase indicator paper. The intensity of the color was analyzed with a color scanner. Amniotic glucose was 27.1 +/- 1.6 mg/dl on day 10 and continued in this range with some fluctuation until day 20 when it decreased. We isolated days 10, 11, and 16 embryonic sites from their decidua and incubated them at 0 degrees C and 38 degrees C while measuring glucose concentration. The glucose concentration did not show significant decrease at 0 degrees C on day 10 or 11 or on day 16 at 38 degrees C. At 38 degrees C the day 10 embryo amniotic fluid glucose disappeared after 22 minutes and the day 11 amniotic fluid glucose was gone in 34 minutes. These depletion times were statistically different. The magnitude of glucose depletion on day 10 was shown by calculation to be approximately 323 mumoles/gm protein per hour which is a substantial portion of the glucose utilized by the embryo as determined in previous experiments (731 mumoles/gm protein per hour). This model may serve as a way to study glucose utilization by embryos after their exposure to various teratogens.

Amniotic Fluid↗