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PubMed2000

The influence of bowel atresia in gastroschisis on fetal growth, cardiotocograph abnormalities and amniotic fluid staining.

OBJECTIVES: To investigate the relationship between intestinal atresia and fetal growth, intrapartum cardiotocograph abnormalities and staining of the amniotic fluid in gastroschisis pregnancies. DESIGN: Observational study of 115 gastroschisis pregnancies conceived between 1980 and 1996. SETTING: South West Region of England. MAIN OUTCOME MEASURES: Birthweight, intrapartum cardiotocograph abnormality, staining of amniotic fluid and Apgar score. RESULTS: There was no statistically significant difference between the Apgar scores or the frequency of amniotic fluid staining of gastroschisis cases with or without atresia. Patent bowel gastroschisis was associated significantly with more cardiotocograph abnormalities (61% vs 26%, P = 0.026) and reduced growth (P = 0.02), when compared with cases with intestinal atresia. CONCLUSIONS: Bile vomiting may be an important cause of amniotic fluid staining. Bowel atresia protects against the increased incidence of cardiotocograph and growth abnormality found in cases of gastroschisis which may reflect their aetiology.

Amniotic Fluid↗
PubMed2007

Amniotic fluid induces rapid epithelialization in the experimentally ruptured fetal mouse palate--implications for fetal wound healing.

Cleft of the secondary palate is one of the most common congenital birth defects in humans. The primary cause of cleft palate formation is a failure of fusion of bilateral palatal shelves, but rupture of the once fused palate has also been suggested to take place in utero. The possibility of post-fusion rupture of the palate in humans has hardly been accepted, mainly because in all the cleft palate cases, the cleft palatal edge is always covered with intact epithelium. To verify whether the intrauterine environment of the fetus plays roles in wound healing when the once fused palate is torn apart, we artificially tore apart fetal mouse palates after fusion and cultivated them in culture medium with or without mouse or human amniotic fluid. We thereby found that the wounded palatal edge became completely covered with flattened epithelium after 36 hours in culture with amniotic fluid, but not in culture without amniotic fluid. Using histological and scanning electron microscopic analyses of the healing process, it was revealed that the epithelium covering the wound was almost exclusively derived from the adjacent nasal epithelium, but not from the oral epithelium. Such actions of amniotic fluid on the fetal wound were never simulated by exogenous epidermal growth factor (EGF), albumin, or both. In addition, the rapid epithelialization induced by amniotic fluid was not prevented by either PD168393 (an inhibitor of the EGF receptor-specific tyrosine kinase) or SB431542 (a specific inhibitor of TGFbeta receptor type I/ALK5). The present study provides new insights into the unique biological actions of amniotic fluid in the repair of injured fetal palate.

Activin Receptors, Type I↗
PubMed1994

[Intrauterine defensive mechanism of amniotic fluid and fetal membranes].

To determine the intrauterine defensive role of urinary trypsin inhibitor (UTI), we studied the effects of UTI in amniotic fluid, fetal membranes and myometrium. The level of UTI was 94 +/- 34 U/ml in neonatal urine (compared to adult urine 8.0 +/- 6.0 U/ml) and 88 +/- 37 U/ml in amniotic fluid. This may indicate that the main source of UTI in the amniotic fluid is the fetal urine. UTI was found to be concentrated in vernix, fetal intestine, amniotic membranes and uterine myometrium. Immunostaining of term amnion revealed a dark staining for UTI, whereas in premature deliveries UTI staining was markedly decreased. In myometrium, the concentration of UTI was found to be increased during pregnancy compared to non pregnant myometrium. Also, placentas were well stained for UTI in term pregnancy. Thus UTI has an important role in amniotic fluid, fetal membranes, placenta and uterine muscles. UTI has an inhibitory effect on several enzymes and cytokines. UTI was found to inhibit neutrophil elastase activity as well as trypsin activity. Its inhibitory activity was increased in the presence of lipid. LPS stimulated amnion cells trapped more UTI than unstimulated amnion cells. UTI in amnion cells was released after addition of 1% meconium solution. UTI was also found to inhibit the effect of IL-1, TNF and interleukin-8 on amnion. These results indicate that UTI localized in amnion is important in the protection of fetal membrane especially against bacterial infections and cytokines. It is known that endothelin (ET), prostaglandin F2 alpha (PGF2 alpha) and oxytocin can induce uterine contraction. UTI could inhibit uterine contractions stimulated by ET, PGF2 alpha and oxytocin in isometric contraction test. UTI could also inhibit cervical maturation induced by interleukin-8. Therefore UTI is essential for maintenance of pregnancy. From the isometric contraction tests, we assumed that UTI might works through regulation of calcium entry or availability in the cells. Initial increase in intracellular calcium was also inhibited by UTI pre incubation dose dependantly. We examined the change in intracellular calcium at single cell level by digital image analysis with Fura 2AM as a calcium probe. At resting level UTI incubation did not produce any significant changes in intracellular free calcium. Thrombin, LPS, interleukin-8 and ET-1, known calcium agonists could increase intracellular calcium in fibroblasts, amnion and uterine myocytes. Whereas as the same doses of those known calcium agonists could not change the intracellular free Ca2+ concentrations in UTI pre incubated fibroblasts, amnion cells and uterine smooth muscle cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Abortion, Threatened↗
PubMed1984

Stable isotope dilution analysis of 3-hydroxyisovaleric acid in amniotic fluid: contribution to the prenatal diagnosis of inherited disorders of leucine catabolism.

A quantitative assay for 3-hydroxyisovaleric acid in amniotic fluid was developed using D6-3-hydroxyisovaleric acid as an internal standard. 3-Hydroxyisovaleric acid was isolated by liquid partition chromatography and the amount determined by selected ion monitoring, ammonia chemical ionization gas chromatography-mass spectrometry of the trimethylsilyl derivatives. The concentration of 3-hydroxyisovaleric acid in ten normal amniotic fluid was 4.52 +/- 1.73 mumol/l. The level was elevated eight-fold in the amniotic fluid from a pregnancy resulting in the birth of a child with biotin-responsive multiple carboxylase deficiency. The stable isotope dilution assay of 3-hydroxyisovaleric acid in amniotic fluid is a rapid, sensitive and accurate method for the prenatal diagnosis of this disorder, and may be of value in the prenatal diagnosis of other inherited disorders of leucine catabolism.

Amino Acid Metabolism, Inborn Errors↗
PubMed1980

Diagnosis of fetal death in utero with amniotic fluid creatine kinase.

There is a need for a widely available and accurate method of diagnosing fetal death in utero. The accepted standard technique for making this diagnosis is real-time ultrasonography, but this procedure is not available at all hospitals. However, almost every hospital laboratory has the capability of measuring creatine kinase in body fluids. One hundred eight pregnant women underwent real-time ultrasonography and analysis of amniotic fluid for creatine kinase. Ninety-one women with conditions that required amniocentesis during the second and third trimesters had a life fetus by ultrasonographic examination and a low level of amniotic fluid creatine kinase (0 to 3 sigma units per milliliter). Seventeen women with a dead fetus by ultrasonographic examination had an abnormally high level of amniotic fluid creatine kinase (5 to 9,800 sigma units per milliliter). The only erroneous diagnosis in this series involved a set of twins, one of whom was shown by real-time ultrasonography to be alive and the other dead. Both sacs contained abnormally high, although different, levels of creatine kinase. Analysis of creatine kinase in amniotic fluid is a reliable technique for diagnosing fetal death.

Amniotic Fluid↗
PubMed2001

Amniotic fluid and hemodynamic model in monochorionic twin pregnancies and twin-twin transfusion syndrome.

We developed a mathematical model of monochorionic twin pregnancies and twin-twin transfusion syndrome (TTTS), combining both fetal fluid dynamics and fetoplacental growth and circulation alterations and assuming that transplacental fluid flow from mother to fetus accounts for normal fetal and amniotic fluid volumes. Ten coupled differential equations, describing fetal total body and amniotic fluid volumes, their osmolalities, and fetal blood colloid osmotic pressure, for both donor and recipient twins, were solved numerically. Amniotic flows are controlled by fetal plasma osmolality and hydrostatic and colloid osmotic pressures. We included varying placental anastomoses and placental sharing of the circulations. Consistent with clinical experience, model predictions are: fetofetal transfusion from unidirectional arteriovenous anastomoses cause oligo-polyhydramnios, a normal size recipient but hypovolemic donor; compensating oppositely directed deep and superficial anastomoses moderate discordant development; and anhydramnios results from mild and severe TTTS, where milder forms may even present earlier in gestation than severe TTTS. Unequal placental circulatory sharing may exacerbate discordant development. In conclusion, our model simulates a wide variety of realistic manifestations of amniotic fluid volume and fetal growth in TTTS related to placental angioarchitecture. The model may allow an assessment of the efficacy of current therapeutic interventions for TTTS.

Amniotic Fluid↗
PubMed1989

[Changes in 13,14-dihydro-15-keto-prostaglandin F2 alpha and dehydroepiandrosterone-sulfate (DHA-S) levels in amniotic fluid and maternal peripheral venous blood in late pregnancy and during labor].

To study the role of prostaglandin (PG) and steroids on the mechanism of parturition, levels of 13,14-dihydro-15-keto-prostaglandin F2 alpha (dhk.PGF2 alpha) and dehydroepiandrosterone-sulfate (DHA-S) in amniotic fluid and plasma during late pregnancy, labor, and puerperium, were measured by radioimmunoassay and gas chromatography-mass spectrometry. Seven patients at term were selected for this study and labor was induced by amniotomy. Amniotic fluid and maternal peripheral blood were obtained simultaneously from each individual at A) the artificial rupture of the membranes before the onset of labor, B) the onset of labor, C) the second stage of labor, D) the delivery, and E) 2 hours postpartum. No increase in either amniotic fluid or plasma dhk-PGF2 alpha was noticed when compared before and at the onset labor. However, there was a steady increase during labor and the maximum was reached at delivery. The dhk-PGF2 alpha concentration in amniotic fluid correlated well with that in maternal plasma. A similar pattern was observed in the DHA-S levels both in amniotic fluid and plasma. No apparent change was observed before the onset of labor, but there was a tendency to increase during labor. A significant correlation between the levels of dhk.PGF2 alpha and DHA-S in amniotic fluid was noticed. From the results obtained above, it is suggested that PG and DHA-S may be involved not in the initiation of labor but in the promotion of parturition. The biological significance of DHA-S on PG levels remains to be clarified.

Amniotic Fluid↗
PubMed1979

The peroxidase content and the antibacterial activity of the amniotic fluid.

A possible relationship between the antibacterial activity of amniotic fluid and its peroxidase content was examined. Antibacterial activity, assessed by counting colonies of S. aureus following 24 hour incubation, was present in 76% of the samples studied. It was not related to gestational age. Peroxidase activity, assessed by the O-dianisidine method, was not found in any of the amniotic fluid samples examined.

Amniotic Fluid↗
PubMed1985

Zinc and alpha-fetoprotein in amniotic fluid from early pregnancies with fetal malformations.

Zinc and alpha-fetoprotein concentrations were quantitated in second-trimester amniotic fluid from 111 normal pregnancies and 29 pregnancies with various fetal malformations. The zinc level varied between 0.3 and 4.0 mumol/L (median 1.3) in the normal samples and between 1.8 and 17.9 mumol/L (median 5.0) in cases with malformations. The level was above 2.5 times the normal median in 23 of the 29 (79%) gestations with the malformations. Amniotic fluid zinc and alpha-fetoprotein levels showed a positive correlation (r = 0.78), and the alpha-fetoprotein level was elevated in all gestations with fetal disorders and elevated zinc levels, as well as in five of the six cases with fetal defects but normal zinc concentrations. The elevation of zinc was more marked than that of alpha-fetoprotein in three cases of fetal malformations. Five samples were false positive on alpha-fetoprotein assay but contained normal zinc levels. Removal of alpha-fetoprotein from amniotic fluid had no effect on the zinc concentration, indicating that zinc is not bound to amniotic fluid alpha-fetoprotein.

Adult↗
PubMed1992

The disposition of valproate and its metabolites in the late first trimester and early second trimester of pregnancy in maternal serum, urine, and amniotic fluid: effect of dose, co-medication, and the presence of spina bifida.

We have studied 52 pregnancies in epileptic women taking long-term valproate and have measured the concentrations of the parent compound and 13 of its metabolites by gas chromatography-mass spectrometry in amniotic fluid, maternal serum, and 24 h maternal urine samples. All metabolites of valproate present in the serum could also be detected in the amniotic fluid, although at much lower concentrations. Amniotic fluid concentrations of valproate and several of its metabolites ((E) delta 2-valproate, (2E,3'E) delta 2,3'-valproate, and 3-keto-valproate) correlated with total valproate concentrations as well as with unbound valproate concentrations in maternal serum. We suggest that the amniotic fluid acts as a deep compartment, with slow appearance and disappearance of valproate and its main metabolites. The data further suggest that during the first and early second trimesters of pregnancy the beta-oxidation of valproate decreases. In pregnancies associated with fetal neural tube defects (n = 5) significantly higher daily doses of valproate were used compared with normal pregnancies (n = 47). This resulted in higher concentrations of valproate in maternal serum. However, the metabolite patterns in maternal serum, 24 h urine samples, and amniotic fluid did not show any significant differences in pregnancies with neural tube defects.

Amniocentesis↗
PubMed1990

Ferning of amniotic fluid contaminated with blood.

The diagnosis of rupture of the membranes by traditional methods of nitrazine paper determination and fern crystallization has been reported to be 93-98% accurate. Although the presence of contaminants in amniotic fluid has been reported to reduce this accuracy, the literature is conflicting concerning these effects. We examined the effect of blood contamination on the accuracy of diagnosing ruptured membranes. Amniotic fluid was obtained from 38 patients between 16-38 weeks' gestational age who underwent amniocentesis. Amniotic fluid was immediately mixed with freshly obtained, heparinized fetal cord blood in varying concentrations (blood to amniotic fluid 1:5, 1:10, 1:20). The slides were examined microscopically for the presence of ferning. All samples were fern-positive, but many had atypical ferns described as "skeletonized." As the concentration of blood to amniotic fluid increased, the number of atypical ferns increased (32 of 38 at 1:5, 22 of 38 at 1:10, and nine of 38 at 1:20). We conclude that the presence of blood may alter the morphology of the fern, but does not act as a contaminant that would affect the accuracy of the test.

Amniotic Fluid↗
PubMedDate not supplied

Maternal serum and amniotic fluid hydroxyproline levels in neural tube defects.

OBJECTIVE: The present study was aimed to evaluate the alterations of maternal serum and amniotic fluid OH- proline levels in neural tube defects (NTD). METHODS: 38 patients and 23 control subjects were selected from the patients followed up by the Departments of Genetics and Antenatal Care, Zekai Tahir Burak Women's Health, Education and Research Hospital. Amnion fluid and blood samples were synchronously obtained from NTD and control subjects. OH-proline levels were detected spectrophotometrically. RESULTS: Amniotic fluid OH-proline levels were found to be increased in NTD patients compared to control subjects (p < 0.05). There was no statistically significant difference in serum OH-proline levels between the groups. CONCLUSION: The results showed an increase in amniotic fluid OH-proline levels in NTD compared to the control group. It may be suggested that the increased OH-proline levels reflect increased collagen turnover in NTD patients. Indeed, amniotic fluid OH-proline levels may be a candidate as an additional diagnostic parameter in NTD.

Amniotic Fluid↗
PubMed1981

[Causes of the antibacterial action of amniotic fluid (author's transl)].

The activity of E. coli, Group B Streptococci, Strept. faecalis, Stap. epidermidis and Staph. aureus in human amniotic fluid was studied. In comparison to Standard I broth an inhibited growth of test organisms was observed. The bacteriostatic effect of amniotic fluid was most significant with Staph. epidermidis. We further investigated the causes for this bacterial growth inhibition with Staph. epidermidis. We showed that it is independent from the initial number of bacteria as well as from zinc and phosphate content, the phosphate-to-zinc ratio and from the protein content in amniotic fluid samples. It could neither be influenced by heating nor by autoclaving, but was immediately lost by adding Standard I both to amniotic fluid samples. In the concentrations we investigated the bacteriostatic effect was independent from the influence of steroid hormones.

Amniotic Fluid↗
PubMed2001

Sonographic assessment of amniotic fluid volume between 11 and 24 weeks of gestation: construction of reference intervals related to gestational age.

OBJECTIVE: At present, most of the methods for sonographic assessment of amniotic fluid volume are unreliable in the second trimester of pregnancy, or else they do not present nomograms related to gestational age. DESIGN: The aim of this prospective cross-sectional study was to construct normal reference ranges of four ultrasound parameters for the evaluation of amniotic fluid volume which could be applied in the second trimester. For these parameters we calculated normal curve limits suitable for use in clinical practice. SUBJECTS: From a population of normal pregnant women between the 12th and the 24th weeks of gestation undergoing a routine ultrasound examination during 1997 at our institute, 273 were found to be suitable for the study, after the exclusion of all cases which presented any feto-maternal pathology or complications up to the 24th week. METHODS: The largest 'amniotic pocket' in a vertical direction, free of small fetal parts and umbilical cord, was measured: the maximum vertical and transverse diameters were measured on the same scan; the mean diameter and the product of the two diameters were calculated. The 'mean amniotic fluid diameter', the 'two-diameter pocket', the 'largest vertical pocket' and the 'largest transverse pocket' were the four sonographic parameters considered. RESULTS: The four parameters correlated well with gestational week and with the biparietal diameter; the normal reference intervals and normal curve were then calculated. All these parameters were found to have good intra- and interoperative reproducibility. CONCLUSIONS: We conclude that the use of an ultrasound semiquantitative method based on the measurement of a single amniotic fluid pocket and involving normal reference intervals according to gestational age could improve the early diagnosis of amniotic fluid variations during the second trimester, although this has yet to be confirmed by extensive clinical trials.

Amniotic Fluid↗
PubMed1989

Variation in some enzymes in amniotic fluid and maternal serum during pregnancy.

The following 10 enzymes were assayed in 187 amniotic fluid and maternal serum samples at 15-42 weeks of gestation: alkaline phosphatase, heat-stable alkaline phosphatase (only in amniotic fluid), acid phosphatase, alanine aminotransferase, aspartate aminotransferase, alpha-amylase, gamma-glutamyltransferase, creatine kinase, lactate dehydrogenase, and lysozyme. The normal reference ranges are reported for amniotic fluid and maternal serum enzymes, together with the abnormal values accompanying neural tube defects and EPH-gestosis. The determination of gamma-glutamyltransferase, heat-stable alkaline phosphatase and creatine kinase was found to be of appreciable diagnostic significance in clinical practice.

Amniotic Fluid↗
PubMed1986

Use of parenteral antibiotic therapy to eradicate bacterial colonization of amniotic fluid in premature rupture of membranes.

The successful eradication of bacterial colonization of the amniotic fluid with parenteral maternal antibiotic therapy in a preterm gestation with premature rupture of membranes is reported. Amniotic fluid, obtained by amniocentesis, of a pregnancy complicated by premature rupture of membranes at 29 weeks was found to have three different bacterial species (Bacteroides bivius, Veillonella parvula, and Peptococcus). Because of prematurity and clinical quiescence, parenteral antibiotics were administered to the mother. Repeat amniocentesis was performed during treatment and 48 hours after the cessation of antibiotics failed to demonstrate bacterial growth in the amniotic fluid. Fetal lung maturity was attained, and an induction of labor was performed when antepartum heart rate testing was equivocal. This was accomplished without maternal or fetal/neonatal mishap.

Adult↗
PubMed2001

Maternal or fetal origin of rhesus monkey (Macaca mulatta) amniotic fluid leukocytes can be identified by polymerase chain reaction using the zinc finger Y gene.

Leukocytes can be found in substantial numbers within the intrauterine tissues and amniotic fluid of women, and play a central role in the pathophysiology of infection-related preterm labor by their production of proinflammatory mediators. It remains unclear whether these leukocytes represent a fetal immune response, a maternal response, or a combination of the two. The objective of this study was to develop a test in the rhesus monkey (Macaca mulatta) suitable for determining the percentage of male fetal cells present in a population of leukocytes recovered from blood or amniotic fluid. We found inadequate specificity for rhesus monkey cells using commercial human Y-chromosome paint kits (fluorescence in situ hybridization (FISH)). Human-specific primers for the repetitive Y chromosome DYZ-1 locus employed in the polymerase chain reaction (PCR) produced an unacceptable percentage of false positives. However, we successfully developed a PCR-based test using rhesus-specific primers for the zinc finger Y (ZFY) locus. Densitometry of PCR products from known ratios of male and female adult peripheral leukocytes generated a linear standard curve which provided quantitative results and required only 400 cells per sample. The rhesus beta globin (RBG) gene served as an internal control. The PCR test correctly discriminated the sex of peripheral leukocytes in 20 adult males, 20 adult females, two male fetuses, and one female fetus. Serial samples of amniotic fluid from four chronically catheterized rhesus monkeys bearing male fetuses were used to confirm the utility of this assay for quantifying fetal cells in amniotic fluid. In conclusion, we have developed a PCR test which is suitable for distinguishing male from female cells in adult and fetal blood and in amniotic fluid, which lends itself to a variety of diagnostic and biologic applications in the rhesus monkey and potentially in other nonhuman primates.

Amniotic Fluid↗