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Stable isotope dilution analysis of galactitol in amniotic fluid: an accurate approach to the prenatal diagnosis of galactosemia.

A stable isotope dilution assay for galactitol in amniotic fluid has been developed using selected ion monitoring chemical ionization gas chromatography-mass spectrometry of the hexaacetate derivative. [1,1-2H2]Galactitol was synthesized for use as the internal standard. Galactitol is a component of normal amniotic fluid with a mean concentration of 0.70 +/- 0.18 mumol/liter (n = 5). The amniotic fluid of a fetus with galactosemia had a concentration of 7.96 mumol/liter. Mannitol, sorbitol, and inositol were also found to be normal constituents of amniotic fluid. This stable isotope dilution assay is a rapid accurate method for measurement of galactitol in amniotic fluid for prenatal diagnosis of galactosemia.

Amniotic Fluid↗

Effects of intrauterine growth restriction and intraamniotic insulin-like growth factor-I treatment on blood and amniotic fluid concentrations and on fetal gut uptake of amino acids in late-gestation ovine fetuses.

OBJECTIVES: To investigate, in the late-gestation ovine fetus: 1) amino acid concentrations in blood and amniotic fluid, 2) the effects of intrauterine growth restriction (IUGR) induced by placental embolization on these concentrations, 3) fetal gut uptake of glutamine in healthy and IUGR fetuses, and 4) the effects of intraamniotic insulin-like growth factor-I (IGF-1) treatment on these parameters. METHODS: Fetuses were randomly assigned to control (n = 9), IUGR + saline (n = 9), or IUGR + IGF-1 (n = 11) groups. IUGR was induced by uteroplacental embolization from 114 to 119 days (term = 145 days). IUGR fetuses received daily intraamniotic injections of saline or IGF-1 (20 microg/d) from 120 to 130 days. RESULTS: Baseline amino acid concentration was higher in fetal blood than amniotic fluid for all essential amino acids except lysine and histidine, but was lower for serine, alanine, and methylhistidine. Embolization reduced total amino acid concentration in blood and amniotic fluid by approximately 15%. Concentrations were reduced for serine, glutamine, and methylhistidine in blood and for serine in amniotic fluid, but were increased for glycine, alanine, and asparagine in blood and for alanine in amniotic fluid. Glutamine was taken up by the fetal gut (glutamine:oxygen quotient of 0.65) and citrulline was released by the gut. IGF-1 treatment did not alter amino acid concentration in blood or amniotic fluid, but reduced gut uptake of glutamine from blood and the gut glutamine:oxygen quotient by 15%. Citrulline release was unchanged. CONCLUSIONS: These data suggest that amniotic fluid amino acids are not simply filtered from fetal blood and may provide an important pool of nutrients for the fetus. They demonstrate for the first time that glutamine is taken up by the fetal gut. IGF-1 treatment may promote gut utilization of amino acids from the amniotic fluid pool.

Amino Acids↗

Growth-factor-like substance in amniotic fluid in the rat: effect on the development of fetal colonic goblet cells.

Extra- and intrauterine fetuses were studied to explore the effect of amniotic fluid on the development of colonic goblet cells. Significant differences in the density of goblet cells in developing colon were observed during fetal days 19-22 between these two groups, suggesting that amniotic fluid affects the development of the colonic goblet cells, especially during the period between days 19 and 21 when rat fetuses began to swallow amniotic fluid, as demonstrated by injecting India ink into the amniotic cavity. By immunological dot blot analysis, the amniotic fluid and gastric juice from intrauterine fetuses were positive for epidermal growth factor (EGF), whereas maternal abdominal serous fluid and gastric juice from extrauterine fetus were negative for EGF. The present results indicate that the amniotic fluid during late gestation greatly affects the development of the colonic goblet cells in rat fetuses and that the trophic factors for the cells seems to be EGF or an EGF-like substance in the amniotic fluid.

Abdomen↗

[Antiphagocytic activity of human amniotic fluid].

The preincubation of PMN leukocytes in amniotic fluid obtained at the end of the gestation period resulted in decreased values of phagocytic activity (about 37%), phagocytic index (about 21.5%) and mainly of the candidacidal capacity to C. albicans (about 72%). The level of lysozyme (11.8 +/- 7.2 mg.l-1) and the level of alpha 1-antitrypsin (0.745 +/- 0.53 g.l-1) were also determined in amniotic fluid. The level of alpha 2-macroglobulin and the proteolytic activity at acidic and mildly alkaline pH could not be established by the methods applied. (Fig. 1, Ref. 23.)

Amniotic Fluid↗

Exposure to amniotic fluid inhibits closure of open wounds in the fetal rabbit.

In many species, open cutaneous fetal wounds do not heal in utero. Such open wounds have been shown to close only after their exclusion from amniotic fluid, thus leading to the hypothesis that amniotic fluid inhibits open wound healing. Therefore the effect of amniotic fluid exposure on the healing of open fetal skin wounds was studied. Fetuses of New Zealand White rabbits received a full-thickness circular 4 mm diameter skin punch biopsy wound. Wounds were left uncovered, covered with a latex patch, or covered with a latex patch with a central hole (doughnut). This third group provided for wound exposure to amniotic fluid while controlling for any wound splinting effect of the patch. Wounds were harvested after 5 days, the wound area was determined planimetrically, and wound edges were examined by means of light microscopy. Analysis of glycosaminoglycans in the wound extra-cellular matrix was performed on a separate group of wounds treated similarly. Uncovered wounds enlarged by an average of 60%, whereas wounds covered with the doughnut patch enlarged by an average of 24%. In contrast, the wounds in the patch-covered group decreased in size by an average of 84%. Histologically all groups contained proliferating fibroblasts and epithelial migration at the wound edge but also an absence of granulation tissue. The patch-covered wounds, which had decreased wound area, were significantly enriched in hyaluronic acid. These results suggest that the healing of the patch-covered wounds occurs without the formation of granulation tissue, presumably through a process of cellular migration and proliferation and that healing was inhibited by exposure to amniotic fluid. Hyaluronic acid has been shown to be permissive of cellular migration and to play a key role in tissue regeneration. Therefore, we speculate that direct exposure of open wounds to amniotic fluid during the late stages of fetal development in the rabbit prevents hyaluronic acid deposition, which in turn may alter wound closure.

Journal Article↗

Correlation of maternal plasma volume and composition with amniotic fluid index in normal human pregnancy.

OBJECTIVE: Early in human pregnancy, maternal plasma osmolality decreases and plasma volume begins to increase. Continued plasma volume expansion is linked to fetal fluid acquisition, and ultimately normal amniotic fluid volume. Although the mechanisms for these physiological responses are uncertain, we hypothesized that the time course of maternal plasma hypotonicity, plasma volume expansion and amniotic fluid index (AFI) were interrelated. We sought to examine these interrelationships during human pregnancy. METHODS: Normal primiparous pregnant patients had monthly determinations of plasma volume using Evans blue from the second trimester to term, and 8 weeks after delivery. At each visit, fasting, morning blood samples were also drawn to measure hematocrit, osmolality, electrolytes, urea, glucose and creatinine. AFI was determined by ultrasound. RESULTS: Plasma volume, blood volume and AFI demonstrated significant increases with advancing gestational age. There were no significant changes in plasma sodium or osmolality from the second trimester to term. AFI was not associated with either plasma volume or osmolality. CONCLUSION: Although similar factors may therefore be responsible for the initiation of plasma volume expansion and plasma hypotonicity, the magnitude and degree of change of these processes are probably under independent regulation. Individual patient variation in AFI is a result of factors other than maternal blood volume or composition.

Amniotic Fluid↗

Sonographic estimation of amniotic fluid volume. Subjective assessment versus pocket measurements.

Amniotic fluid volume (AFV) estimation is an important part of routine obstetric sonography. Despite the clinical importance placed upon excessive or diminished AFV in pregnancy, there is little uniformity in the way it is estimated sonographically. We compared AFV estimations obtained using two commonly employed sonographic methods "subjective" visual assessment and amniotic fluid pocket measurements. Estimates obtained using both methods correlated closely. In addition, there was excellent intraobserver and interobserver agreement among estimates obtained using subjective criteria. This supports the belief that experienced observers tend to agree on the sonographic appearance of normal, excessive or decreased AFV. Methods for AFV estimation and potential pitfalls are discussed.

Amniotic Fluid↗

Increased matrix metalloproteinase activity and reduced tissue inhibitor of metalloproteinases-1 levels in amniotic fluids from pregnancies complicated by premature rupture of membranes.

OBJECTIVES: It has been suggested that increased matrix metalloproteinases activity promotes the weakening of the amniochorion during normal and premature rupture of membranes. This study was designed to determine whether levels of matrix metalloproteinases and the tissue inhibitor of metalloproteinases-1 in amniotic fluid change in a pattern consistent with this hypothesis. STUDY DESIGN: Gelatinolytic activity, measured by a soluble substrate assay and zymography, and the concentrations of tissue inhibitor of metalloproteinases-1 were estimated in amniotic fluid obtained from (1) normal early gestations, (2) normal term pregnancies with labor, (3) normal term pregnancies without labor, and (4) pregnancies complicated by premature rupture of membranes. The 92 kd type IV collagenase (matrix metalloproteinase-9) was also detected in amniotic fluid by Western blotting. RESULTS: Matrix metalloproteinase activities were higher in amniotic fluid from normal term pregnancies with labor and pregnancies complicated by premature rupture of membranes than from early pregnancies and term gestations without labor. The amniotic fluid from term pregnancies with labor or pregnancies with premature rupture of membranes contained several gelatinases, as revealed by zymography. The major amniotic fluid gelatinolytic activity in premature rupture of membranes and term pregnancies with labor corresponded to matrix metalloproteinase-9. Tissue inhibitor of metalloproteinases-1 concentrations were highest in early-pregnancy amniotic fluid, followed by term gestation with labor, term gestation without labor, and premature rupture of membranes. CONCLUSIONS: Normal labor and premature rupture of membranes are associated with increased levels of matrix metalloproteinases, particularly matrix metalloproteinase-9 in amniotic fluid. Premature rupture of membranes is associated with reduced levels of tissue inhibitor of metalloproteinases-1. The imbalance between matrix metalloproteinases and tissue inhibitor of metalloproteinases-1 in amniotic fluid may reflect a disorder that promotes premature rupture of membranes.

Amniotic Fluid↗

Sonographic detection of echogenic amniotic fluid and its clinical significance.

It has been suggested that meconium-stained amniotic fluid can be detected in the antepartum period by means of ultrasound, based on the following findings: (1) a diffuse echogenic pattern throughout the amniotic cavity, (2) a clear contrast between the amniotic fluid and the umbilical cord, and (3) layering in the more dependent areas. Reported is a case in which a similar picture was obtained in a third trimester, high-risk pregnancy, but representing vernix instead of meconium. We are aware of six cases, including ours, of sonographic detection of echogenic amniotic fluid. The review of these cases indicates that the findings refer more often to a considerable amount of vernix rather than meconium, and it is not necessarily associated with postmaturity. If echogenic amniotic fluid is identified in an obstetrical ultrasound, we recommend: (1) rule-out meconium by amniocentesis or amnioscopy, or (2) study fetal well-being by means of nonstress test and/or biophysical profile. Thus, a false positive diagnosis of fetal distress in these cases could be easily avoided.

Amniotic Fluid↗

Is amniotic fluid material in the central circulation of peripartum patients pathologic?

Cytologic findings of amniotic fluid material (AFM) in pulmonary arterial blood (PAB) of survivors of amniotic fluid embolism (AFE) are assumed to be pathologic. However, no cytologic studies of central blood from patients without clinical AFE have been reported. To address this question PAB samples from peripartum patients without clinical AFE were examined for the presence and extent of AFM (including squames, mucin, and lanugo hair). Ten samples were obtained from five patients. All patients had at least one sample postpartum. Peripheral blood from a nonpregnant adult female control was processed similarly. Results were compared to a PAB sample from a patient with clinical AFE. The patient with clinical AFE had many squames, clumps of lanugo hair, and mucin in one sample. In six of ten study samples, there were squames, accompanied in two cases by lanugo hair or trophoblast. In nine of ten samples there was mucin. There appeared to be no difference in cytologic findings in patients according to mode of delivery or sampling time. The control blood sample was negative for amniotic fluid-like material. AFM may be found in peripartum patients without clinical amniotic fluid embolism. A quantitative difference was seen between the index patient and each of our five study patients. These findings suggest that there is a quantitative continuum of AFM transported to the central circulation in peripartum patients which may, in part, explain the varied clinical presentations and severity of AFE.

Adult↗

Noradrenaline to adrenaline ratio in amniotic fluid as an index of fetal maturity.

Amniotic fluid lecithin to sphingomyelin ratio (L/S) and adrenaline concentration, noradrenaline concentration, and noradrenaline to adrenaline ratio (NA/A) were measured in 50 normal women late in pregnancy. Between gestation weeks 37 and 43 the maturity-linked increase in L/S ratio was accompanied by similar increases in NA/A ratio and noradrenaline but by decreases in adrenaline concentration. Although the amniotic fluid NA/A ratio reflects fetal sympathetic nervous system development rather than pulmonary development as mirrored by the L/S ratio, a high correlation existed between log NA/A and L/S, indicating that the NA/A ratio could provide a further index of fetal maturity.

Amniotic Fluid↗

Isolation of pulmonary pressor substances from bovine amniotic fluid.

The pulmonary pressor activity of bovine amniotic fluid resides in substances that appear to be non-polar lipids as suggested by the various chemical and chromatographic procedures. In preliminary experiments, it was found that the pressor activities of the hexane extracts were blocked by prior administration of aspirin to the calf (1 mg/kg), suggesting that pressor effect was mediated via the prostaglandin system. The amniotic fluid lipids, however, differed from the known prostaglandins in being less polar as indicated by thin layer chromatography and in possessing greater pressor activity. It is possible, of course, that the lipids with pulmonary pressor activity participate in the metabolism of the prostaglandins.

Amniotic Fluid↗

Amniotic fluid composition in normal baboon pregnancies.

In order to compare the chemical composition of baboon and human amniotic fluid, 70 amniotic fluid samples were obtained from 58 normal baboon pregnancies at times ranging from 88 to 178 days (term, 180 days). Protein content averaged (+/- S.E.) 0.67 +/- 0.26 gm/dl, osmolality averaged 263 +/- 1.1 mOsm/kg, and pH averaged 7.74 +/- 0.034 units. These variables were not affected by length of gestation. Creatinine content increased parabolically with duration of gestation (r = 0.74; p less than 0.001). Forty percent of the samples had delta OD450 values of 0; the others had delta OD450 values ranging from 0.002 to 0.145. No decline in delta OD450 with gestational maturity was evident. These data suggest that the chemical environment of the baboon fetus is similar but not identical to that of the human fetus.

Amniotic Fluid↗

[Study of interleukin-6 and tumor necrosis factor-alpha levels in maternal serum and amniotic fluid in patients with preterm premature rupture of membrane].

OBJECTIVE: To study the change and clinical significance of maternal serum and amniotic fluid interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) levels in patients with chorioamnionitis and premature rupture of membrane (PROM). METHODS: Twenty normal term-pregnant women as control, and fourty-six women with PROM were enrolled for study. Maternal serum and amniotic fluid IL-6, TNF-alpha levels were measured by radioimmunoassay and enzyme-labeled immunosorbent assay (ELISA). Chorioamnionitis was diagnosed histologically. RESULTS: The maternal serum IL-6 levels and amniotic fluid IL-6, TNF-alpha levels were higher in PROM group than that of control (P < 0.01). There was significant positive correlation between maternal IL-6, amniotic fluid IL-6, TNF-alpha levels and the duration from rupture of membrane, (the longer was time, the higher were the maternal IL-6 and the amniotic fluid IL-6. TNF-alpha). There were 12 cases with chorioamnionitis in patients with PROM and their maternal and amnotic fluid IL-6, TNF-alpha levels were higher than that of non-chorioamnionitis (P < 0.01-0.05). CONCLUSION: IL-6 and TNF-alpha levels in maternal serum and amniotic fluid are valuable clinical indices for identification of chorioamnionitis in patients with PROM.

Adult↗

Amniotic fluid concentrations of renin and aldosterone during development in the fetal sheep.

Simultaneous plasma renin activity and aldosterone levels were measured in the chronically catheterized fetal lamb and pregnant ewe between 100 days gestation and term (140 to 150 days). Amniotic fluid renin levels were not statistically different after the addition of sheep renin substrate [0.76 +/- 0.18 ng/ml/hr (mean and S.E.) under 120 days and 1.33 +/- 0.29 after 125 days]. Amniotic fluid renin levels were unmeasurable without the addition of renin substrate. Basal plasma renin activity levels increased from 4.27 +/- 0.66 ng/ml/hr (P less than 0.01) (mean and S.E.) between 95 to 120 days of gestation in the fetal lamb to 10.14 +/- 1.46 between 125 to 150 days; these levels were greater than the amniotic fluid levels (P less than 0.001). Basal plasma renin activity levels in the pregnant ewe increased from 2.61 +/- 0.40 ng/ml/hr under 120 days of gestation to 7.13 +/- 1.41 (P less than 0.01) after 125 days; these levels were greater than the simultaneous amniotic fluid levels (P less than 0.001). The basal amniotic fluid, fetal, and maternal aldosterone levels of 5.0 +/- 0.53, and 5.83 +/- 0.76 ng/dl (mean and S.E.) increased to 8.97 +/- 0.86, 6.67 +/- 1.21, and 12.28 +/- 2.87 ng/dl, respectively, (P less than 0.05). Neither the fetal nor the maternal levels were statistically different from the amniotic fluid aldosterone levels during development. These data suggest that active renin and renin substrate pass poorly into the amniotic fluid; however, aldosterone from both the fetus and mother pass easily and contribute to the level of this steroid in amniotic fluid.

Aldosterone↗

Selenium status in pregnancy: studies in amniotic fluid from normal pregnant women.

Selenium concentrations were determined in amniotic fluid samples obtained from 111 healthy normal pregnant women (median age, 27.5 years) between 12 and 42 weeks of gestation using hydride generation technique coupled with atomic absorption spectroscopy. There was a gradual decrease in the amniotic fluid selenium concentration with the progress of pregnancy. The negative correlation between the gestational age and amniotic fluid selenium concentration was highly significant (p less than 0.001). The implications of these findings in normal pregnancy are discussed.

Amniotic Fluid↗

The association of birthweight with maternal and cord serum and amniotic fluid growth hormone and insulin levels, and with neonatal and maternal factors in pregnant women who delivered at term.

AIM: To investigate the influence of maternal and cord serum and amniotic fluid growth hormone (GH) and insulin and other neonatal and maternal factors on birthweight. METHODS: A total of 160 pregnant women at 38-42 weeks' gestation were studied. All infants were categorized as small for gestational age (SGA) (n = 50), large for gestational age (LGA) (n = 50) or average for gestational age (AGA) (n = 60). GH and insulin levels were measured in maternal and cord serum and amniotic fluid at birth. RESULTS: GH levels in maternal and cord serum and amniotic fluid showed no differences among the three weight groups (P > 0.05). The cord insulin level was significantly lower in SGA (P < 0.01). The insulin level in venous cord blood correlated with birth and placental weights and neonatal height, whereas maternal serum and amniotic fluid insulin levels, and maternal and cord serum and amniotic fluid GH levels did not show any correlation with birthweight. The cord GH level at birth was correlated with GH levels after 4 postnatal weeks in the SGA group (P < 0.01). In addition, birthweight showed a correlation with prepartum maternal weight, maternal weight gain, maternal height, neonatal length and placental weight in all three weight groups. CONCLUSIONS: Cord GH, maternal serum and amniotic fluid GH and insulin levels did not correlate with birthweight in all three weight groups. The lack of correlation for GH levels in maternal and cord serum and amniotic fluid suggests that these compartments may be non-communicating separate units.

Adult↗

Prolactin in amniotic fluid: its correlation with maternal plasma prolactin.

Prolactin concentrations in amniotic fluid from 319 women with normal pregnancies and 29 women with complicated pregnancies were determined by radioimmunoassay. Prolactin levels varied from 36 ng/ml to 1800 ng/ml mean +/- S.D. = 408 +/- 297) in the normal pregnancy group but showed no definite pattern of rise or fall during pregnancy. No difference in levels was found in complicated pregnancies. Prolactin concentrations in the plasma from 203 of these women were also assayed. The levels in the amniotic fluid were about 9 fold higher than those in the plasma. There was no significant correlation between amniotic fluid and plasma levels of prolactin.

Amniotic Fluid↗