A circadian clock entrained by melatonin is ticking in the rat fetal adrenal.

The adrenal gland in the adult is a peripheral circadian clock involved in the coordination of energy intake and expenditure, required for adaptation to the external environment. During fetal life, a peripheral circadian clock is present in the nonhuman primate adrenal gland. Whether this extends to the fetal adrenal gland like the rat is unknown. Here we explored in vivo and in vitro whether the rat fetal adrenal is a peripheral circadian clock entrained by melatonin. We measured the 24-h changes in adrenal content of corticosterone and in the expression of clock genes Per-2 and Bmal-1 and of steroidogenic acute regulatory protein (StAR), Mt1 melatonin receptor, and early growth response protein 1 (Egr-1) expression. In culture, we explored whether oscillatory expression of these genes persisted during 48 h and the effect of a 4-h melatonin pulse on their expression. In vivo, the rat fetal adrenal gland showed circadian expression of Bmal-1 and Per-2 in antiphase (acrophases at 2200 and 1300 h, respectively) as well as of Mt1 and Egr-1. This was accompanied by circadian rhythms of corticosterone content and of StAR expression both peaking at 0600 h. The 24-h oscillatory expression of Bmal-1, Per-2, StAR, Mt1, and Egr-1 persisted during 48 h in culture; however, the antiphase between Per-2 and Bmal-1 was lost. The pulse of melatonin shifted the acrophases of all the genes studied and restored the antiphase between Per-2 and Bmal-1. Thus, in the rat, the fetal adrenal is a strong peripheral clock potentially amenable to regulation by maternal melatonin.

Melatonin exerts direct inhibitory actions on ACTH responses in the human adrenal gland.

In nonhuman primates and rodents, melatonin acting directly on the adrenal gland, inhibits glucocorticoid response to ACTH. In these species, an intrinsic adrenal circadian clock is involved in ACTH-stimulated glucocorticoid production. We investigated whether these findings apply to the human adrenal gland by determining i) expression of clock genes in vivo and ii) direct effects of melatonin in ACTH-stimulated adrenal explants over a) expression of the clock genes PER1 (Period 1) mRNA and BMAL1 [Brain-Muscle (ARNT)-like] protein, ACTH-induced steroidogenic acute regulatory protein (StAR), and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) and b) over cortisol and progesterone production. Adrenal tissue was obtained from 6 renal cancer patients undergoing unilateral nephrectomy-adrenalectomy. Expression of the clock genes PER1, PER2, CRY2 (Cryptochrome 2), CLOCK (Circadian Locomotor Output Cycles Kaput) and BMAL1, was investigated by RT-PCR in a normal adrenal and in an adenoma. In independent experiments, explants from 4 normal adrenals were preincubated in culture medium (6 h) followed by 12 h in: medium alone; ACTH (100 nM); ACTH plus melatonin (100 nM); and melatonin alone. The explants' content of PER1 mRNA (real-time PCR) and StAR, 3ß-HSD, BMAL1 (immuno slot-blot), and their cortisol and progesterone production (RIA) were measured. The human adrenal gland expresses the clock genes PER1, PER2, CRY2, CLOCK, and BMAL1. ACTH increased PER1 mRNA, BMAL1, StAR, and 3ß-HSD protein levels, and cortisol and progesterone production. Melatonin inhibited these ACTH effects. Our study demonstrates, for the first time, direct inhibitory effects of melatonin upon several ACTH responses in the human adrenal gland.

Cryptochrome 2 expression level is critical for adrenocorticotropin stimulation of cortisol production in the capuchin monkey adrenal.

Timely production of glucocorticoid hormones in response to ACTH is essential for survival by coordinating energy intake and expenditure and acting as homeostatic regulators against stress. Adrenal cortisol response to ACTH is clock time dependent, suggesting that an intrinsic circadian oscillator in the adrenal cortex contributes to modulate the response to ACTH. Circadian clock gene expression has been reported in the adrenal cortex of several species. However, there are no reports accounting for potential involvement of adrenal clock proteins on cortisol response to ACTH. Here we explored whether the clock protein cryptochrome 2 (CRY2) knockdown modifies the adrenal response to ACTH in a primate. Adrenal gland explants from adult capuchin monkey (n = 5) were preincubated for 6 h with transfection vehicle (control) or with two different Cry2 antisense and sense probes followed by 48 h incubation in medium alone (no ACTH) or with 100 nm ACTH. Under control and sense conditions, ACTH increased cortisol production, whereas CRY2 suppression inhibited ACTH-stimulated cortisol production. Expression of the steroidogenic enzymes steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase at 48 h of incubation was increased by ACTH in control explants and suppressed by Cry2 knockdown. Additionally, we found that Cry2 knockdown decreased the expression of the clock gene brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein (Bmal1) at the mRNA and protein levels. Altogether these results strongly support that the clock protein CRY2 is involved in the mechanism by which ACTH increases the expression of steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase. Thus, adequate expression levels of components of the adrenal circadian clock are required for an appropriate cortisol response to ACTH.

Clock gene expression in adult primate suprachiasmatic nuclei and adrenal: is the adrenal a peripheral clock responsive to melatonin?

The circadian production of glucocorticoids involves the concerted action of several factors that eventually allow an adequate adaptation to the environment. Circadian rhythms are controlled by the circadian timing system that comprises peripheral oscillators and a central rhythm generator located in the suprachiasmatic nucleus (SCN) of the hypothalamus, driven by the self-regulatory interaction of a set of proteins encoded by genes named clock genes. Here we describe the phase relationship between the SCN and adrenal gland for the expression of selected core clock transcripts (Per-2, Bmal-1) in the adult capuchin monkey, a New World, diurnal nonhuman primate. In the SCN we found a higher expression of Bmal-1 during the h of darkness (2000-0200 h) and Per-2 during daytime h (1400 h). The adrenal gland expressed clock genes in oscillatory fashion, with higher values for Bmal-1 during the day (1400-2000 h), whereas Per-2 was higher at nighttime (about 0200 h), resulting in a 9- to 12-h antiphase pattern. In the adrenal gland, the oscillation of clock genes was accompanied by rhythmic expression of a functional output, the steroidogenic enzyme 3beta-hydroxysteroid dehydrogenase. Furthermore, we show that adrenal explants maintained oscillatory expression of Per-2 and Bmal-1 for at least 36 h in culture. The acrophase of both transcripts, but not its overall expression along the incubation, was blunted by 100 nm melatonin. Altogether, these results demonstrate oscillation of clock genes in the SCN and adrenal gland of a diurnal primate and support an oscillation of clock genes in the adrenal gland that may be modulated by the neurohormone melatonin.

Maternal melatonin effects on clock gene expression in a nonhuman primate fetus.

In the adult mammal the circadian system, which allows predictive adaptation to daily environmental changes, comprises peripheral oscillators in most tissues, commanded by the suprachiasmatic nucleus (SCN) of the hypothalamus. The external environment of the fetus is provided by its mother. In primates, maternal melatonin is a candidate to entrain fetal circadian rhythms, including the SCN rhythms of metabolic activity. We found in the 90% of gestation capuchin monkey fetus expression of the clock genes Bmal-1, Per-2, Cry-2, and Clock in the SCN, adrenal, pituitary, brown fat, and pineal. Bmal-1, Per-2, and the melatonin 1 receptor (MT1) showed a robust oscillatory expression in SCN and adrenal gland, whereas a circadian rhythm of dehydroepiandrosterone sulphate was found in plasma. Maternal melatonin suppression changed the expression of Bmal-1, Per-2, and MT1 in the fetal SCN. These effects were reversed by maternal melatonin replacement. In contrast, neither maternal melatonin suppression nor its replacement had effects on the expression of Per-2 and Bmal-1 or MT1 in the fetal adrenal gland or the circadian rhythm of fetal plasma dehydroepiandrosterone sulphate. Our data suggest that maternal melatonin is a Zeitgeber for the fetal SCN but probably not for the adrenal gland.

Fetectomy alters maternal pituitary-adrenal function in pregnant rhesus macaques.

The interplay between the fetus and mother may play a key role in the regulation of primate pregnancy and parturition. This study was designed to test the hypothesis that fetectomy alters maternal pituitary-adrenal function. Between 117 and 122 days of gestation (term = 167 days), six rhesus macaques underwent surgery for catheter implantation. At surgery the fetuses were removed while the membranes and placenta were left in situ. Six additional intact catheterized pregnant animals served as controls. Animals were maintained under a 12L:12D cycle with lights-on from 0700 to 1900 h. Beginning at least 1 wk after surgery, maternal arterial blood samples were collected at 3-h intervals for 24 h for hormone and catecholamine analysis. This sampling protocol was repeated at weekly intervals until cesarean delivery at 151-157 days of gestation. Following fetectomy, plasma ACTH, dehydroepiandrosterone sulfate (DHEAS), and cortisol levels were significantly lower (36%, 35%, and 44%, respectively) compared with control animals (P;lt 0.05). Despite a significant reduction in overall levels, the rhythm in maternal plasma cortisol was maintained following fetectomy. Plasma dopamine and norepinephrine were also depressed (P;lt 0.05), whereas epinephrine remained unaffected. Our data clearly demonstrate the role of the fetus in the regulation of the maternal pituitary-adrenal axis during gestation. This interaction plays a significant role in the regulation of maternal endocrine function that may influence the initiation of labor.

Twenty-four-hour pattern of cortisol in the human fetus at term.

OBJECTIVES: Indirect evidence suggests that adrenal steroid production in the human fetus may have a circadian rhythm. To assess whether there is a 24-hour rhythm of fetal cortisol in the human fetus, we investigated the relationship between fetal and maternal cortisol and cortisone concentrations in maternal, umbilical arterial, and umbilical venous blood samples over a 24-hour period. STUDY DESIGN: Elective cesarean sections were scheduled every 2 hours around the clock in 57 term (38-41 weeks' gestation) nonlaboring pregnant women. Plasma cortisol and cortisone concentrations were measured by high-pressure liquid chromatography. RESULTS: The mean 24-hour cortisol concentration was higher in umbilical arterial than in umbilical venous blood samples, 63.6 +/- 4.6 ng/mL (SEM) versus 48.7 +/- 3.2 ng/mL, respectively (P <.05). Fetal plasma cortisol showed a rhythm in the umbilical artery (acme from noon to 4 PM ) (1-way analysis of variance and least significant difference test; P <.05) but not in the umbilical vein. Umbilical arteriovenous differences showed no net transfer of cortisol to the fetus at any time of the day and net fetal production of cortisol from 8 AM to 6 PM. There was limited transfer of cortisone to the fetus and only in the 2 AM -to-noon time interval. CONCLUSION: These data suggest the presence of a 24-hour rhythm of fetal adrenal cortisol secretion that may be controlled by a fetal circadian pacemaker.

The development of circadian rhythms in the fetus and neonate.

The circadian time-keeping system is the neural system that allows predictive adaptation of individuals to the reproducible 24-hour day/night alternations of our planet. A biological clock, the suprachiasmatic nucleus, receives environmental information and imposes a circadian pattern to physiological functions. Since the suprachiasmatic nucleus develops early in gestation and circadian rhythms are present in the fetus and newborn, the circadian system seems to be functional in fetal life and can receive circadian inputs through the mother. The neonate moves to an environment in which the main time giving signal is the light:dark cycle. Teleologically, a term newborn should be fit to face this challenge. But this may be quite different for a preterm infant that trades the circadian environment to which it was previously exposed for the timeless environment of the Neonatal Intensive Care Nursery. Scientists and physicians should seek new experimental and clinical approaches to answer the challenging questions of perinatal chronomedicine.

An oxytocin receptor antagonist (atosiban) in the treatment of preterm labor: a randomized, double-blind, placebo-controlled trial with tocolytic rescue.

OBJECTIVES: This study was designed to evaluate the efficacy and safety of the oxytocin receptor antagonist atosiban in the treatment of preterm labor. STUDY DESIGN: A multicenter, double-blind, placebo-controlled trial with tocolytic rescue was designed. Five hundred thirty-one patients were randomized to receive, and 501 received, either intravenous atosiban (n = 246) or placebo (n = 255), followed by subcutaneous maintenance with the assigned agent. Standard tocolytics as rescue tocolysis were permitted after 1 hour of either placebo or atosiban if preterm labor continued. The primary end point was the time from the start of study drug to delivery or therapeutic failure. Secondary end points were the proportion of patients who remained undelivered and did not receive an alternate tocolytic at 24 hours, 48 hours, and 7 days. RESULTS: No significant difference was found in the time from start of treatment to delivery or therapeutic failure between atosiban and placebo (median, 25.6 days vs 21.0 days, respectively; P =.6). The percentages of patients remaining undelivered and not requiring an alternate tocolytic at 24 hours, 48 hours, and 7 days were significantly higher in the atosiban group than in the control group (all P < or =.008). A significant treatment-by-gestational age interaction existed for the 48-hour and 7-day end points. Atosiban was consistently superior to placebo at a gestational age of > or =28 weeks. Fourteen atosiban-treated patients and 5 placebo-treated patients were randomized at <24 weeks; the incidence of fetal-infant deaths was higher for the atosiban group at <24 weeks. Maternal-fetal adverse events were similar except for injection-site reactions, which occurred more often with atosiban. CONCLUSIONS: In this trial the treatment of patients in preterm labor with atosiban resulted in prolongation of pregnancy for up to 7 days for those at a gestational age > or =28 weeks, and this occurred with a low rate of maternal-fetal adverse effects. In addition, at a gestational age > or =28 weeks, the infant morbidity and mortality of atosiban-initiated standard care were similar to those with placebo-initiated standard care. Given that all patients in this study were eligible for tocolysis and that, in practice, nearly all patients who are eligible for a tocolytic receive one, the benefit of using atosiban is the placebo-like maternal-fetal side effect profile. These observations support the use of this oxytocin receptor antagonist in the treatment of patients in preterm labor with intact membranes. Efficacy and infant outcome data at <28 weeks are inconclusive.

Maintenance treatment of preterm labor with the oxytocin antagonist atosiban. The Atosiban PTL-098 Study Group.

OBJECTIVES: Patients admitted with an acute episode of preterm labor who respond to early intravenously administered tocolysis remain at risk of having subsequent episodes of preterm labor and preterm delivery. Several pharmacologic agents have been used in an attempt to reduce subsequent episodes of preterm labor, and all are associated with significant side effects. Atosiban, an oxytocin receptor antagonist, is effective in the treatment of an acute episode of preterm labor. This study was designed to compare the efficacy and safety of atosiban with those of placebo maintenance therapy in women with preterm labor who achieved uterine quiescence with intravenous atosiban. STUDY DESIGN: A multicenter, double-blind, placebo-controlled trial was designed for patients in preterm labor who responded to early intravenous treatment with atosiban. Five hundred thirteen patients were randomly assigned to receive maintenance therapy, 252 to receive atosiban, and 251 to receive matching placebo. Maintenance therapy was administered as a continuous subcutaneous infusion, via pump, of 30 microg/min to the end of 36 weeks' gestation. The primary end point was the number of days from the start of maintenance therapy until the first recurrence of labor. A secondary end point was the percentage of patients receiving subsequent intravenous atosiban therapy. RESULTS: The time (median) from the start of maintenance treatment to the first recurrence of labor was 32.6 days with atosiban and 27.6 days with placebo (P =.02). At least one subsequent intravenous atosiban treatment was needed by 61 atosiban patients (23%) and 77 placebo patients (31%). Except for injection site reactions, adverse event profiles of atosiban and placebo were comparable. There were 4 neonatal deaths reported in the atosiban group and 5 in the placebo group after the start of maintenance therapy. Infant outcomes (including birth weight) were comparable between maintenance and treatment groups. CONCLUSIONS: Maintenance therapy with the oxytocin receptor antagonist atosiban can prolong uterine quiescence after successful treatment of an acute episode of preterm labor with atosiban. Treatment was well tolerated.

Preterm labor: placental pathology and clinical correlation.

OBJECTIVE: To determine the relevance of ischemia in the incidence of preterm labor. A second objective was to document perinatal outcomes for patients with preterm labor classified according to its clinical, functional, and pathologic characteristics (infectious, ischemic, mixed, or idiopathic). METHODS: Perinatal outcomes were evaluated for 145 consecutive patients with preterm labor, subdivided into etiologic categories according to clinical, functional (Doppler), and morphologic (placental pathology) characteristics. A group of 44 normal pregnancies delivered at term served as controls. RESULTS: Of the preterm labor group, 28.3% were classified as ischemic, compared with 4.5% of the control group (odds ratio and 95% confidence interval = 8.28 [1.8, 51.8]; P < .05). Compared with the control group, the preterm labor patients who delivered preterm had higher rates of ischemia (31.4% compared with 4.5%; P < .05) and infection (16.1% compared with 2.3%; P < .05). Among the preterm labor group, patients classified in the infectious or ischemic subgroups had a higher rate of preterm delivery (95.0% and 90.2% compared with 73.2%; P < .05), admission to the neonatal intensive care unit (75.0% and 61.0% compared with 40.0%; P < .05), and newborn weight under 1500 g (35.0% and 19.5% compared with 3.7%; P < .05) than the idiopathic subgroup. CONCLUSION: Preterm labor resulting from infection or ischemia is associated with a higher perinatal complication rate than idiopathic preterm labor.

Diurnal changes in light intensity inside the pregnant uterus in sheep.

Penetration of light into the pregnant sheep uterus was studied in 9 ewes, gestational ages 40 to 142 days (term 147 days). Light sensors were placed inside the pregnant horn and over the flank skin overlying the position of the uterine horn. To perform the experiments, the ewes were placed in a study cage outdoors and light sensors were connected to a luxometer. Simultaneous measurements were obtained from the intrauterine and the external sensors in the shade at noon. The amount of light detected inside the uterus increased with gestational age from two lux at 40 days to 51.1 +/- 16.5 (n = 5) lux at 142 days (0.2 and 5.4% of the amount of light detected at the maternal flank). Measurements through the 24 h were done in four pregnant ewes at 142 days gestation under natural photoperiod (13.5 light:10.5 dark). In these experiments, the intensity of intrauterine light changed through the 24 h, reflecting the changes in the intensity of the sunlight. Maximal intrauterine light values were observed at noon, corresponding to 4.7% of incident light. Small but detectable values were observed at 0900 and 1800 h. Our data show that, at mid gestation, light reaches the pregnant uterus and that, at late gestation, changes in intrauterine lighting throughout the 24 h are present reflecting the changes in external daylight. Therefore the sheep fetus is exposed to light-dark transitions at dawn and dusk, and to a peak of light at midday.

Dose ranging study of the oxytocin antagonist atosiban in the treatment of preterm labor. Atosiban Study Group.

OBJECTIVE: To evaluate the minimal effective dose regimen of the oxytocin antagonist atosiban in the treatment of acute preterm labor and the effect of a bolus on uterine activity within the first 2 hours compared with no bolus and the same infusion rate. METHODS: A randomized, double-blind (except the ritodrine group), parallel group, multicenter study compared four different intravenous atosiban regimens (6.5 mg bolus plus 300 micrograms/minute, placebo bolus plus 300 micrograms/minute, 2mg bolus plus 100 micrograms/minute, and 0.6 mg plus 30 micrograms/minute) and intravenous ritodrine with respect to the cessation of uterine contractions for 1 hour or more during infusion, four or fewer contractions per hour in the last hour of therapy, and discontinuation because of adverse experiences. Three hundred two patients were enrolled. RESULTS: The lowest dose of atosiban (0.6 mg plus 30 micrograms/minute) was significantly less effective than ritodrine with respect to cessation of contractions and four or fewer contractions per hour in the last hour of therapy. Other atosiban regimens were comparable to ritodrine, except for the drug discontinuation rate for adverse experiences. Bolus therapy with high-dose atosiban resulted in a significantly greater proportion of patients who stopped contracting within the first 2 hours of treatment (17 of 63) compared with those not receiving a bolus (six of 58, P = .017). Because of adverse experiences, the study drug was discontinued in one of 244 atosiban patients and 15 of 58 ritodrine patients. CONCLUSION: Atosiban's effect on uterine activity in preterm labor was enhanced by bolus infusion and was similar to the effect of ritodrine, but with fewer side effects.

Effect of constant light on fetal and maternal prolactin rhythms in sheep.

A 24-h rhythm of plasma PRL is present in fetal sheep. This rhythm is synchronized to an environmental clue (zeitgeber). We determined whether the light-dark cycle (L:D) is a zeitgeber for the fetal PRL rhythm and, if so, whether the mother might convey this zeitgeber to the fetus. We kept nine ewes (twin pregnancies) in constant light (L:L) and five ewes (singleton) in 14:10 L:D from 110 days gestation. Fetuses and mothers were catheterized at 119 days gestation. Blood samples were taken hourly for 24 h after 16 days under L:L or L:D. A mean 24-h rhythm of PRL was found (by RIA) in fetuses under L:D, but not in those under L:L. However, fetuses under L:L showed individual 24-h PRL rhythms (cosinor analysis) whose acrophases were distributed around the clock. Nonsynchronized rhythms persisted after 23 and 30 days of L:L. Acrophases of PRL rhythms within a set of twins were closer than those between sets, suggesting that twins were responding to a common signal. These findings indicate that the L:D cycle is a zeitgeber for the PRL rhythm in fetal sheep and suggest that the mother might convey the zeitgeber.

Effect of chronic hypoxia on myometrial responsiveness in the pregnant rat.

Mechanisms involving the timing of normal parturition are not well understood in most animal species. To gain a greater understanding of the mechanisms, we employed hypoxia to perturb the normal system of parturition. The present study was designed to investigate the effects of chronic hypoxia on myometrial contractility in the near-term pregnant rat. Rats were exposed to room air (control) or to continuous hypoxia (10.5% O2) either from experimental days 19 through 21 (2-day exposure) or from experimental days 15 through 21 (6-day exposure). On day 21, blood was collected for hormone assays, and the uterine horns were collected from each dam. One horn was snap-frozen in liquid nitrogen for oxytocin (OT) receptor analysis, and the other was used for in vitro assessment of myometrial contractile responses to cumulative doses of OT or arginine vasopressin (AVP). Hypoxic exposure resulted in approximately 60% reduction of the maximal myometrial contractile response to OT and a significant reduction in OT binding sites from 256.9 +/- 34.9 to 84.9 +/- 21.3 fmol/mg protein (P<0.01). In contrast, the contractile response to AVP was unaffected after exposure to chronic hypoxia (P> 0.05). Additionally, we observed no difference in the plasma concentrations of estrogen, progesterone, and corticosterone. We conclude that chronic hypoxia decreased the effectiveness of OT-specific contractile mechanisms, at least partially through a decrease in OT binding sites.

Transvascular albumin transport and protein replenishment after haemorrhage in the chronically catheterized pregnant rabbit.

Pregnancy is characterized by increases in both blood and interstitial volumes, but the mechanisms are unknown. To test the hypotheses that blood volume (BV) recovery after haemorrhage in pregnant (P) is faster than in nonpregnant (NP) rabbits, and that this can be explained by a higher capillary filtration, a 20% BV haemorrhage was produced in a group of 7 P and 7 NP rabbits. We determined the BV recovery (measured by 99Tc), the total plasma protein mass and the haematocrit at 10, 20, 30, 60 and 1440 min after haemorrhage. Arterial and venous pressures and heart rate were measured during a 30-min control period, during the 10 min of haemorrhage, and for the following 70 min. In a separate group of P and NP rabbits, 125I-labelled albumin was infused and blood samples were obtained at 10, 20, and 30 min. Capillary filtration was expressed as the percentage of radioactive albumin that left the intravascular space after 30 min. No differences were found in the rate of BV recovery, arterial and venous pressures, or heart rate response between P and NP animals (P > 0.1). By 24 h total plasma protein mass (TPPM) was significantly increased above baseline in the P rabbits by an average+SEM of 13 + 3.7%, but not in NP rabbits (5 + 4.5%). At 30 min, the amount of labelled albumin in the intravascular space decreased by 15.4 + 3.4% for Pv. 8.2 + 1.2% for NP rabbits (P < 0.05). There was a high correlation between TPPM recovery and BV recovery in the P (r = 0.96) and NP rabbits (r = 0.95). In conclusion, we did not find differences in the rate at which the BV recovered during pregnancy, despite the increased capillary permeability. During pregnancy, the combination of the latter with an increase in the TPPM elevation above pre-haemorrhage levels suggests a faster rate of protein mobilization.

Endothelin-1 potentiates the in vitro contractile response of pregnant human myometrium to oxytocin.

OBJECTIVE: This study was designed to test the hypothesis that endothelin-1 pretreatment of human myometrium at subcontractile doses in vitro will enhance the contractile response to oxytocin. STUDY DESIGN: In vitro contractile oxytocin dose-response curves were generated by use of myometrial strips collected from nonpregnant women (n = 7), pregnant patients at elective cesarean section (n = 7), and patients in active labor (n = 7) in the presence or absence of 10(-9) mol/L endothelin-1. Contractile responses were analyzed by on-line computer, and data were normalized to the maximum response to potassium. RESULTS: Pretreatment with endothelin-1 significantly increased the maximal contractile response of pregnant myometrium (p < 0.01 compared with control). In marked contrast myometrium from nonpregnant patients was unaffected by endothelin-1 pretreatment. Values for the two-point discrimination and Hill coefficient were not different among the treatment groups. CONCLUSION: Endothelin-1 potentiates the oxytocin response of myometrium from pregnant but not nonpregnant women. We speculate that a high circulating level of a uterotonin-like oxytocin may not be necessary to initiate labor. The synergistic interaction between different uterotonins may be sufficient.

Placental passage of the oxytocin antagonist atosiban.

OBJECTIVE: We wanted to determine the degree of placental transfer of atosiban (Antocin), an oxytocin antagonist, in pregnant women at term. We also assessed the effects of the infusion on umbilical cord blood gases at birth and the maternal hematocrit drop after cesarean section. STUDY DESIGN: Eight women undergoing elective cesarean section at term were studied. Each received an infusion of 300 micrograms/min of atosiban over 208 to 443 minutes; the infusion was continued up to the time of cord clamping. Uterine vein and umbilical blood samples were obtained simultaneously. They were assayed by specific radioimmunoassay. Cord blood gases were obtained and compared with those from a control group of women undergoing elective cesarean section. RESULTS: The mean (+/- SD) maternal uterine vein concentration was 331.9 +/- 42.9 ng/ml, compared with 42 +/- 13 ng/ml in the umbilical vein (p < 0.05). The mean maternal/fetal was 12 +/- 0.03, which was not affected by the length of infusion. There was no significant difference in the hematocrit drop between the cesarean delivery groups: 5.9 +/- 0.4 for the control group versus 5.8 +/- 1.1 for the atosiban group (p > 0.1). The mean cord pH was 7.27 for the atosiban group versus 7.27 for the control group (n = 141) (p > 0.1). One year follow-up of the infants (n = 7) was normal. CONCLUSIONS: Our results show minimal placental transfer of atosiban. Drug levels did not increase with longer infusions, and no effect was seen on umbilical cord gases. Administration of atosiban even at high doses up to the time of delivery did not increase maternal blood loss at cesarean section.

Fetectomy alters maternal endocrine and uterine activity rhythms in rhesus macaques during late gestation.

OBJECTIVE: The current study was designed to test the hypothesis that fetectomy will eliminate or substantially alter rhythms in maternal estradiol concentrations and subsequently reduce or eliminate uterine activity rhythms. STUDY DESIGN: Six rhesus macaques underwent surgery for catheter implantation between days 117 and 122 of gestation (term = 167 days). At surgery the fetuses were removed and the membranes and placenta remained intact. Thirteen additional catheterized pregnant animals served as controls. Maternal arterial blood samples were collected for hormone analysis at 3-hour intervals for 24 hours, starting at 9 AM. This sampling protocol was performed four times at weekly intervals until 151 to 157 days' gestation. RESULTS: A significant rhythm (p < 0.01) in estradiol was determined in the control animals with peak concentrations observed in the morning hours whereas the progesterone peak was observed at night. In the fetectomy group mean plasma estradiol concentrations decreased significantly from 312 +/- 34 to 110 +/- 8 pg/ml throughout the study (p < 0.01). Despite a trend toward elevated morning levels, the estradiol rhythm was ablated. The uterine contractile rhythm observed in the control animals with peak activity between 10 PM and midnight (p < 0.01) was also ablated after fetectomy. Basal concentrations of progesterone were significantly lower than control values. CONCLUSIONS: (1) Fetectomy resulted in the elimination of the maternal estradiol rhythm. (2) The uterine activity rhythm was lost after fetectomy. These data suggest that the fetus, by supplying precursors of estrogen, may play an indirect role in the regulation of maternal estradiol rhythms, which in turn appear to play a key role in regulating uterine activity rhythms.

Is uterine blood flow controlled locally or systemically in the pregnant rabbit?

OBJECTIVE: We tested the hypothesis that uterine blood flow is regulated by systemic circulating factors. The alternative hypothesis is that uterine blood flow is regulated by local factors. STUDY DESIGN: Adult female New Zealand White rabbits were subjected to a unilateral tubal ligation and thereafter allowed to become pregnant (n = 9). A group of nonpregnant one-tube-ligated animals served as controls (n = 8). On day 21 of gestation uterine blood flow in the pregnant and nonpregnant uterine horns were measured with 15 microns microspheres. The concentration of prostaglandin E2 metabolites were measured in blood from the uterine veins and from the arterial circulation. RESULTS: Absolute uterine blood flow in the pregnant uterine horn was 12.9 +/- 4.7 versus 5.2 +/- 1.4 ml in the nonpregnant horn (p < 0.05). However, when expressed by blood flow per gram of tissue they were not different (p > 0.1). The uterine blood flow for the nonpregnant uterine horn in the pregnant animals was the same as that of the horns from nonpregnant animals. The level of prostaglandin E metabolites was greater in the uterine vein draining the pregnant horn compared to the nonpregnant horn (p < 0.05). CONCLUSION: These data support the conclusion that the increase in uterine blood flow observed during pregnancy is controlled largely by local factors induced by pregnancy.