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M L Casey

Publications and source records attributed to M L Casey.

At least 73 records · Page 4Linked to original sources
PubMed1990

Myosin light chain phosphorylation in human myometrial smooth muscle cells.

Ca2+/calmodulin-dependent phosphorylation of the 20-kDa regulatory light chain of myosin is of signal importance in the initiation of contraction in a number of smooth muscle tissues. In this investigation, we evaluated the relationship between intracellular free Ca2+/concentration [( Ca2+]i) and the extent of myosin light chain phosphorylation in cultured human myometrial smooth muscle cells. Treatment of myometrial cells with ionomycin caused a concentration- and time-dependent increase in [Ca2+]i and phosphorylation of myosin light chain. Temporally, the increases in light chain phosphorylation and [Ca2+]i in response to ionomycin were similar. In myometrial cells treated with ionomycin (10(-5) M) for 10 s, [Ca2+]i increased from 138 to 800 nM; in these same cells, myosin light chain phosphorylation increased from 5% to a maximum value of 54%. Half-maximal phosphorylation of myosin light chain was attained at 300 nM [Ca2+]i. Treatment of myometrial smooth muscle cells with prostaglandin (PG) F2 alpha (10(-8) M) and PGE2 (10(-8) M) caused a proportionate increase in [Ca2+]i and myosin light chain phosphorylation. In addition, [Ca2+]i and myosin light chain phosphorylation increased in response to oxytocin and angiotensin II. These findings indicate that a number of uterotonic agents effect an increase in [Ca2+]i, which in turn causes phosphorylation of myosin light chain. Furthermore, the concentration of Ca2+ in the cytoplasm is a primary determinant for myosin light chain phosphorylation in human myometrial smooth muscle cells.

Angiotensin II↗
PubMed1989

Inactivation of prostaglandins in human decidua vera (parietalis) tissue: substrate specificity of prostaglandin dehydrogenase.

Prostaglandin dehydrogenase catalyzes the initial reaction in the inactivation of prostaglandin E2 and F2 alpha. To address the potential importance of this enzyme in regulating the tissue levels of active prostaglandins, we evaluated the kinetic properties of prostaglandin dehydrogenase in uterine decidua vera tissue of women. Specifically, we characterized the enzyme activity under optimal in vitro conditions in cytosolic fractions of uterine decidua vera tissue obtained at term and compared the substrate and cosubstrate specificities of prostaglandin dehydrogenase in cytosolic fractions of decidual tissues. The incubation conditions were optimized with either prostaglandin E2 or F2 alpha and nicotinamide-adenine dinucleotide or nicotinamide-adenine dinucleotide phosphate as substrates to ensure linearity of product formation with time of incubation and protein concentration. The apparent Michaelis-Menten constant of nicotinamide-adenine dinucleotide-dependent prostaglandin dehydrogenase for prostaglandin E2 was 5.5 mumol/L. The apparent Michaelis-Menten constant of nicotinamide-adenine dinucleotide phosphate-dependent prostaglandin dehydrogenase for prostaglandin F2 alpha was 15 mumol/L. Prostaglandin E2 serves as a better substrate for prostaglandin dehydrogenase than does prostaglandin F2 alpha, irrespective of the cosubstrate. In cytosolic fractions of decidual tissues, the specific activity (apparent Vmax) of nicotinamide-adenine dinucleotide-dependent prostaglandin dehydrogenase was greater than that of nicotinamide-adenine dinucleotide phosphate-dependent prostaglandin dehydrogenase. In addition, we found that in decidual tissue obtained before or after the onset of labor, the specific activity of prostaglandin dehydrogenase varied widely. In tissues obtained after delivery by cesarean section, no significant differences were apparent in the specific activity of the enzyme before (9.3 to 125.8 nmol/min/mg protein) and after (27.8 to 103.4 nmol/min/mg protein) the onset of labor. In cytosolic fractions of decidual tissue obtained after vaginal delivery, the specific activity of nicotinamide-adenine dinucleotide-dependent prostaglandin dehydrogenase ranged from undetectable levels to 38.4 nmol/min/mg protein. We speculate that nicotinamide-adenine dinucleotide-dependent prostaglandin dehydrogenase in decidua serves to regulate the levels of bioactive prostaglandins in decidua vera tissue and the amounts of prostaglandins (and metabolites) produced in decidua or fetal membranes that reach myometrium and fetal membranes and enter maternal blood and amniotic fluid.

Cesarean Section↗
PubMed1989

Transforming growth factor-beta inhibits prostaglandin production in amnion and A431 cells.

We studied the effect of transforming growth factor-beta (TGF-beta) on prostaglandin E2 (PGE2) production and mitogenesis in human amnion cells and compared the response in amnion cells with that in A431 cells. Both amnion cells and A431 cells respond to epidermal growth factor (EGF) with increased production of PGE2 whereas EGF promotes mitogenesis in amnion cells but not in A431 cells. In amnion cells, TGF-beta was not mitogenic, and did not alter the mitogenic response of cells to EGF. Treatment of amnion cells with TGF-beta did, however, cause a decrease in PGE2 production relative to untreated cells, although EGF stimulated PGE2 production was not attenuated. In A431 cells, TGF-beta acted to decrease PGE2 production relative to untreated cells and to attenuate the stimulation of PGE2 production effected by EGF. The inhibitory action of TGF-beta on PG production in amnion and A431 cells is contrary to the stimulation of PG production in mouse calvaria reported by others and is suggestive that the effect of TGF-beta on prostaglandin production, like its effect on growth, varies between different cell types. Inhibition of PG production by treatment of amnion or A431 cells with mefenamic acid did not alter thymidine incorporation into DNA in response to EGF; similarly, the addition of PGE2 or PGF2 alpha to culture media of amnion or A431 cells had no effect on mitogenesis (in the absence or presence of EGF). Based on these findings, we conclude that PG production and EGF action on proliferation (stimulation in amnion cells; inhibition in A431 cells) are dissociated.

Amnion↗
PubMed1989

Cachectin/tumor necrosis factor-alpha formation in human decidua. Potential role of cytokines in infection-induced preterm labor.

This study was conducted as part of an investigation to evaluate the hypothesis that bacterial toxins (LPS or lipoteichoic acid), acting on macrophage-like uterine decidua to cause increased formation of cytokines, may be involved in the pathogenesis of infection-associated preterm labor. We found that cachectin/tumor necrosis factor-alpha (TNF-alpha) was synthesized and secreted into the culture medium by human decidual cells and explants in response to treatment with LPS. LPS treatment also caused an increase in PGF2 alpha production by decidual cells and explants. In amnion cells in monolayer culture, TNF-alpha stimulated PGE2 formation, and TNF-alpha was cytostatic (inhibited [3H]thymidine incorporation into DNA) but not cytolytic in amnion cells. TNF-alpha was not detectable (less than 0.34 ng/ml) in the amniotic fluid of normal pregnancies at midtrimester or at term before or after the onset of labor (n = 44); but TNF-alpha was present at concentrations between 2.8 and 22.3 ng/ml in amniotic fluids of 4 of 20 pregnancies with intact membranes complicated by preterm labor (less than 34 wk gestational age). LPS was present in 10 of the 20 amniotic fluids of preterm labor pregnancies, including all four in which TNF-alpha was present. Bacteria were identified in only one of the four LPS-positive, TNF-alpha-positive fluids. Cytokine formation in macrophage-like decidua may serve a fundamental role in the pathogenesis of preterm labor, including increased prostaglandin formation and premature rupture of the membranes.

Amniotic Fluid↗
PubMed1988

Epidermal growth factor stimulation of prostaglandin E2 biosynthesis in amnion cells. Induction of prostaglandin H2 synthase.

Amnion is believed to be a tissue of signal importance, anatomically and functionally, in the maintenance of pregnancy and during the initiation of parturition. Epidermal growth factor (EGF)-like agents cause a striking increase in the secretion of prostaglandin E2 (PGE2) in human amnion cells but only if arachidonic acid is present in the culture medium. To investigate the regulation of arachidonic acid metabolism by EGF-like agents in amnion, we used mEGF and human amnion cells in primary monolayer culture as a model system. The amount of PGE2 secreted into the culture medium was quantified by radioimmunoassay and the rate of conversion of [14C]arachidonic acid to [14C]PGE2 (PGH2 synthase activity) in cell sonicates was determined under optimal in vitro conditions. Treatment of amnion cells with mEGF led to a marked increase in the rate of production of PGE2. The specific activity of PGH2 synthase (viz. the combined activities of prostaglandin endoperoxide (PGH2) synthase and PGH2-PGE isomerase) was increased by 2-5-fold in cells treated with mEGF. Treatment of amnion cells with mEGF for 4 h did not affect the specific activities of phospholipase A2 or phosphatidylinositol-specific phospholipase C. By immunoisolation of newly synthesized, [35S]methionine-labeled PGH2 synthase, we found that mEGF stimulated de novo synthesis of the enzyme. Thus, mEGF acts in human amnion cells in primary monolayer culture to increase the rate of PGE2 biosynthesis by a mechanism that involves induction of PGH2 synthase; the manifestation of EGF action on PGE2 biosynthesis is dependent on the presence of nonesterified arachidonic acid.

Amnion↗
PubMed1988

Assay of bacterial endotoxin (lipopolysaccharide) in human amniotic fluid: potential usefulness in diagnosis and management of preterm labor.

The long-range goal of our research is to determine whether the presence of bioactive agents of infection in amniotic fluid may serve as sensitive indexes of the existence of infection as the cause of preterm labor in a given pregnancy. The aim of this study was to explore the possibility that bacterial endotoxin (lipopolysaccharide) could be detected and quantified in amniotic fluid. In particular, we sought to ascertain (1) if amniotic fluid could be collected in a manner to prevent endotoxin contamination, (2) whether there were inhibitors of lipopolysaccharide in amniotic fluid, and (3) if Limulus amebocyte lysate-based assays could be used to identify and quantify lipopolysaccharide in this fluid. We found that the Limulus amebocyte lysate assays (gelation assay and chromophore generation assay) were useful in the qualitative and quantitative analysis of lipopolysaccharide in amniotic fluids. Lipopolysaccharide-mediated generation of chromophore in the presence of amniotic fluid was accelerated strikingly compared with that of lipopolysaccharide in endotoxin-free water. In three amniotic fluid samples obtained during preterm labor, lipopolysaccharide was detectable in aliquots of 1 to 10 microliters by use of the gelation assay and lipopolysaccharide was quantifiable by use of chromophore generation assays. Two of these amniotic fluid samples were sterile as determined by bacteriologic examination; in the third sample, Fusobacterium species was identified. We suggest that these assays may be extremely useful in the identification of lipopolysaccharide in amniotic fluid. Indeed, lipopolysaccharide may serve as one marker of infection useful in establishing the cause of preterm labor.

Amniotic Fluid↗
PubMed1988

Prostacyclin biosynthesis by cultured human myometrial smooth muscle cells: dependency on arachidonic or linoleic acid in the culture medium.

Myometrial smooth muscle cells in culture were incubated for 18 hours in medium that contained serum (10%); under these conditions, there was a six to 26-fold increase in the amount of 6-keto-prostaglandin F1 alpha that accumulated in the medium (i.e., prostacyclin production) compared with that present after incubation in serum-free medium. In serum-free and serum-containing media, treatment of these cells with dexamethasone (10(-8) mol/L) or cortisol (10(-7) mol/L) suppressed the biosynthesis of prostacyclin by approximately 80% and 64%, respectively. Arachidonic acid (bound to fatty acid-free human serum albumin) added to serum-free medium caused a concentration-dependent increase in the production of prostacyclin by myometrial cells. Arachidonic acid was maximally effective at a concentration of 10(-5) mol/L and caused a five- to 28-fold increase in the biosynthesis and secretion of prostacyclin. Linoleic acid (bound to albumin) in serum-free medium also caused a concentration-dependent increase in the production of prostacyclin; however, the amount of prostacyclin produced in the presence of linoleic acid was lower than that produced in the presence of an equimolar concentration of arachidonic acid. In the presence of arachidonic (10(-5) mol/L) or linoleic acids (10(-4) mol/L) in serum-free medium, the addition of dexamethasone (10(-8) mol/L) or cortisol (10(-7) mol/L) suppressed but did not inhibit completely prostacyclin production. These findings are indicative that arachidonic and linoleic acids in the culture medium support prostacyclin biosynthesis by human myometrial smooth muscle cells. The inhibition of prostacyclin production by glucocorticosteroids in the absence or presence of extracellular arachidonic (or linoleic) acid may be caused by inhibition of phospholipase A2 activity.

6-Ketoprostaglandin F1 alpha↗
PubMed1988

Dehydroepiandrosterone therapeutics: acetylation of DHA in mouse liver.

This investigation was designed to evaluate the possibility that the therapeutic benefits of dehydroepiandrosterone (DHA)-feeding in mice is mediated by (1) a metabolite of DHA formed in liver or (2) by way of the obligatory hepatic metabolism of DHA fed in large amounts. We found that the pattern of metabolism of DHA is strikingly different when DHA in low (tracer) quantities is incubated with mouse liver compared with that found when DHA in high concentrations is incubated with this tissue. In the former case, the principal metabolites are sulfoconjugates and other polar compounds, e.g. hydroxylated products. In the case of DHA metabolism when the substrate is present in high concentrations (1-100 microM), the principal metabolite formed is delta 5-androstenediol. And in this case, there is the formation of very nonpolar metabolites, which we have identified as the acetates of DHA and delta 5-androstenediol. We find further that the acetates are formed by a transacetylation mechanism in which performed acetylated compounds, e.g. pregnenolone acetate and ethyl acetate, can serve directly as co-substrates; but (at least in short-term incubations), Na+-acetate and acetyl CoA do not serve as co-substrates. We suggest that the therapeutic benefits of DHA-feeding in mice may be, in part, the result of alterations in hepatic intermediary metabolism that is obliged by the metabolism of DHA when this otherwise inert agent is fed in large amounts. Thus, DHA-feeding may serve to cause changes similar to those that are beneficial or therapeutic with dietary manipulations including caloric restriction.

Acetylation↗
PubMed1988

Origin of deoxycorticosterone and deoxycorticosterone sulfate in human pregnancy: absence of steroid 21-sulfatase activity in sulfatase-deficient placenta.

The activity of steroid 21-sulfatase, the enzyme that catalyzes the hydrolysis of deoxycorticosterone sulfate (DOC-SO4) is demonstrable in human placenta. Thus, it is possible that this placental enzyme, by way of the hydrolysis of either DOC-SO4 or 21-hydroxypregnenolone mono- or di-sulfate of fetal origin, may be important in the biosynthesis of DOC, which is present in the plasma of pregnant women in high concentration. To investigate this issue further, we evaluated steroid 21-sulfatase activity in microsomal preparations of a sulfatase-deficient placenta. Immediately after delivery, at term, of a living male fetus with sulfatase deficiency, a microsome-enriched fraction of placental tissue was prepared; sulfatase activity was evaluated by use of three substrates, viz. dehydroisoandrosterone sulfate (DS), estrone sulfate (E1-SO4), and DOC-SO4, in various concentrations. Similar incubations were conducted with aliquots of a microsome-enriched fraction prepared from placental tissue of a normal fetus that was delivered, at term, within minutes of the time of delivery of the infant with sulfatase deficiency. In microsomal fractions from the normal placenta, each of the steroid sulfates was hydrolyzed. In the absence of microsomes, and in the presence of microsomal fractions from the sulfatase-deficient placenta, the hydrolysis of DOC-SO4 and DS was not detected. Moreover, in microsomes prepared from the sulfatase-deficient placenta, E1-SO4 was hydrolyzed at a rate that was only 10% of that in incubations with microsomal preparations of the normal placenta. We conclude that with sulfatase deficiency, the placenta is deficient not only in sulfatase activity for steroid-3-sulfates but for steroid 21-sulfates, e.g. DOC-SO4, as well.

17-alpha-Hydroxypregnenolone↗
PubMed1988

Epidermal growth factor, vanadate, and 12-O-tetradecanoylphorbol-13-acetate inhibit growth and stimulate prostaglandin E2 production in A431 cells.

The cell line designated as A431 is characterized by an extraordinary capacity for binding of epidermal growth factor (EGF); but paradoxically, these cells also are characterized by failure of replication in response to treatment with EGF and other mitogens. In this study, we took advantage of this unique response of A431 cells to EGF and other mitogenic agents. Specifically, we investigated the dependency of mitogen-stimulated prostaglandin production on mitogenesis. The effects of treatment of A431 cells with epidermal growth factor and two other mitogenic agents, viz., sodium orthovanadate and 12-O-tetradecanoylphorbol-13-acetate (TPA), on DNA synthesis and prostaglandin production were evaluated. The rate of prostaglandin production in A431 cells that were treated with each of these agents increased in a manner that was dependent upon the duration of treatment and the concentration of the agent tested. Thus, by use of A431 cells, we find that the effects of 'mitogenic' agents on cell replication and prostaglandin production are clearly dissociable.

Cell Line↗
PubMed1987

Biochemical markers of contraction in human myometrial smooth muscle cells in culture.

Phosphorylation of a light chain subunit of myosin by Ca2+ and calmodulin-dependent myosin light chain kinase is believed to be essential for smooth muscle contraction. The biochemical properties of the myosin phosphorylation system in human myometrial smooth muscle cells in monolayer culture were compared with those of human myometrial tissue and nonmuscle cells in culture. Native myosin was isolated from other cellular proteins of crude homogenates by polyacrylamide gel electrophoresis (in the presence of pyrophosphate) and quantified by densitometry. The myosin content of myometrial smooth muscle cells in culture and that of myometrial tissue were similar and four- to five-fold greater than that of human endometrial stromal cells or skin fibroblasts in culture. The specific activities of myosin light chain kinase in homogenates of myometrial smooth muscle cells that were maintained in culture and in myometrial tissue were similar (2.05 +/- 0.18 and 1.60 +/- 0.37 nmol phosphate incorporated per min per mg protein, respectively). On the other hand, enzyme activity in skin fibroblasts was only 5% of that in myometrial smooth muscle cells. Myosin light chain kinase activity in myometrial smooth muscle cells was dependent upon Ca2+ and was inhibited reversibly by the calmodulin antagonist, calmidazolium. The intracellular Ca2+ concentration measured by quin2 fluorescence was 0.12 microM in resting cells and increased in a concentration-dependent manner with KCl to a maximal value of 0.47 microM. These results indicate that biochemical processes important for smooth muscle contraction are retained in human myometrial smooth muscle cells in culture.

Calcium↗
PubMed1987

Mineralocorticosteroids and pregnancy: regulation of extra-adrenal deoxycorticosterone production by estrogen.

During pregnancy, deoxycorticosterone (DOC) in maternal plasma is produced principally by 21-hydroxylation of circulating progesterone in a number of extra-adrenal tissues. Estrogen acts, directly or indirectly, to increase the transfer constant of conversion of progesterone to DOC. In this study, we evaluated the regulation of DOC production during pregnancy by considering the plasma levels of progesterone, the kinetics of extra-adrenal steroid 21-hydroxylase, and the stimulation of the conversion of progesterone to DOC by estrogen.

Anencephaly↗
PubMed1987

Epidermal growth factor-stimulated prostaglandin E2 production in human amnion cells: specificity and nonesterified arachidonic acid dependency.

The production of prostaglandin E2 (PGE2) by human amnion cells in primary monolayer culture was stimulated 2- to 150-fold by mouse epidermal growth factor (mEGF). The effect of mEGF on amnion cell PGE2 production was dependent on the time of treatment and the concentration of mEGF used. The rate of PGE2 production by these cells was maximal between 2 and 4 h of treatment with mEGF; thereafter, the rate of production of PGE2 declined. The stimulation of PGE2 production was maximal at concentrations of mEGF of greater than 5 ng/ml. A number of other growth factors, steroid and protein hormones, and various other test agents were ineffective or only minimally in stimulating PGE2 production by amnion cells. The stimulatory effect of mEGF on PGE2 production in these cells was dependent on the presence of serum in the culture medium; alternatively, mEGF was effective in stimulating PGE2 production by amnion cells in serum-free medium that was supplemented with arachidonic acid bound to albumin. Thus, we conclude that the marked stimulation of PGE2 production by amnion cells treated with mEGF is not due to an action of mEGF to stimulate the release of arachidonic acid from cellular glycerophospholipid storage forms; rather, these data are supportive of the conclusion that mEGF-stimulated PGE2 production is dependent on the presence of nonesterified, i.e., free, arachidonic acid in the medium.

Amnion↗
PubMed1986

Separation and quantitation of fatty acids by high-performance liquid chromatography.

To facilitate the determination of the fatty acid composition of tissues and the investigation of fatty acid metabolism, we developed a method for the rapid separation by high-performance liquid chromatography and quantitation (by ultraviolet light absorption) of p-bromophenyl esters of fatty acids which vary in chain length from 10 to 22 carbon atoms. The utility of the method was demonstrated by evaluating the fatty acid composition of human uterine decidua vera tissue and human endometrial stromal cells that are maintained in monolayer culture.

Chromatography, High Pressure Liquid↗
PubMed1986

Response of human amnion cells in culture to 1,25-dihydroxycholecalciferol: increased 25-hydroxycholecalciferol 24-hydroxylase activity and prostaglandin E2 formation.

We investigated the possibility that 1,25-dihydroxycholecalciferol acts in human amnion as a physiologic calcium ionophore to effect increased prostaglandin E2 production in this tissue. This possibility was based on the propositions that this metabolite of vitamin D3 acts in other target tissues, that is, bone, intestine, chorioallantoic membrane, kidney, and intestine, to effect increased calcium absorption, that 1,25-dihydroxycholecalciferol is present in amniotic fluid, which bathes the amnion, and that the mobilization of calcium in amnion is associated with stimulation of enzymes, namely, phospholipase A2 and phosphatidylinositol-specific phospholipase C that act ultimately to effect the release of arachidonic acid, which in turn leads to increased prostaglandin E2 production. We found that human amnion cells, maintained in primary monolayer culture, are responsive to 1,25-dihydroxycholecalciferol. Treatment of these cells with 1,25-dihydroxycholecalciferol for 18 hours brought about the induction, in a dose-dependent manner, of 25-hydroxycholecalciferol 24-hydroxylase, a marker of 1,25-dihydroxycholecalciferol action. In addition, treatment of human amnion cells in monolayer culture with 1,25-dihydroxycholecalciferol caused, within 12 hours, a significant increase in the synthesis of prostaglandin E2, which was maintained for the duration of treatment, that is, 48 hours. These findings may be indicative of a significant physiologic role for 1,25-dihydroxycholecalciferol in metabolic processes that are important in amnion, including transport, amniotic fluid volume homeostasis, and the initiation of parturition.

Amnion↗
PubMed1986

Responsiveness of human carcinoma cells of gynecologic origin to 1,25-dihydroxycholecalciferol.

In responsive tissues the induction of 25-hydroxycholecalciferol 24-hydroxylase activity is a marker of 1,25-dihydroxycholecalciferol action. In this study we investigated the responsiveness of various malignant cells, which were derived from carcinomas of gynecologic origin, to 1,25-dihydroxycholecalciferol; we did so because it has been demonstrated that replication of certain malignant human cells that are 1,25-dihydroxycholecalciferol-responsive is inhibited by treatment with this substance. We found that 1,25-dihydroxycholecalciferol treatment caused an increase in 25-hydroxycholecalciferol 24-hydroxylase activity in gynecologic carcinoma cells that were maintained in monolayer culture. The cells were established from ovarian, endometrial, cervical, and vaginal neoplasms. After treatment with 1,25-dihydroxycholecalciferol (10 nmol/L) for 18 hours, the specific activity of 25-hydroxycholecalciferol 24-hydroxylase increased between 2,4- and 49-fold in these cells. This increase in 25-hydroxycholecalciferol 24-hydroxylase activity is suggestive that certain gynecologic malignancies are responsive to 1,25-dihydroxycholecalciferol. We also found, however, that 1,25-dihydroxycholecalciferol did not act to decrease the rate of replication of these cells; indeed in ovarian adenocarcinoma cells, 1,25-dihydroxycholecalciferol caused a decrease in doubling time; in the other cells of this study, there was no effect of 1,25-dihydroxycholecalciferol on cell replication.

Adenocarcinoma↗
PubMed1986

The effect of fetal urine on arachidonic acid metabolism in human amnion cells in monolayer culture.

Human amnion cells in primary monolayer culture were used as a model system to evaluate the regulation of arachidonic acid metabolism and prostaglandin E2 production in amnion. Amnion cells were incubated with carbon 14-labeled arachidonic acid, and after various times the distribution of radiolabeled arachidonic acid in the lipids of these cells were determined. After incubation for 72 hours, 91% of the total radiolabeled arachidonic acid incorporated into cellular lipids was present in glycerophospholipids, and 9% was present in neutral lipids. The formation of [14C]prostaglandin E2 from [14C]arachidonic acid was maximal after 8 hours. In these studies the effect of human fetal urine on arachidonic acid metabolism in these cells was investigated (the production of prostaglandin E2 by amnion cells is increased by treatment with fetal urine). In cells incubated with [14C]arachidonic acid in the incubation medium, treatment with fetal urine for 4 hours caused a threefold to fourfold increase in [14C]prostaglandin E2 production, yet there were no detectable differences in the content of [14C]arachidonic acid in specific glycerophospholipids or neutral lipids of the cells after such treatment. In other studies, amnion cells were preincubated for 72 hours with [14C]arachidonic acid, and thereafter the cells were treated with fetal urine for 24 hours. With fetal urine treatment the amount of [14C]arachidonic acid in triacylglycerols decreased significantly compared with that in nontreated cells, but the formation of [14C]prostaglandin E2 was not increased. Thus we suggest that in response to fetal urine, prostaglandin E2 is formed from arachidonic acid that is released from a highly specific, stimulus-sensitive lipid pool or else from arachidonic acid that is derived from extracellular sources.

Amnion↗