Hepatic histopathologic condition does not correlate with laboratory abnormalities in HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count). Our objective was to categorize the histologic findings in the liver in patients with HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count) and to correlate these findings with the severity of clinical laboratory abnormalities. Eleven patients with laboratory criteria for HELLP syndrome who required cesarean delivery underwent needle biopsy of the liver under direct visualization. Eight patients had periportal hemorrhage, and six had fibrin deposition. Fatty infiltration was seen in four, one with large-droplet fat, three with microvesicular fat. There was no statistically significant correlation between the severity of the histologic findings of periportal hemorrhage and fibrin deposition and the clinical laboratory findings. Fatty infiltration did not correlate with the severity of the HELLP syndrome's histologic condition, but, in contrast, did correlate with thrombocytopenia and aminotransferase elevations. Laboratory abnormalities do not accurately reflect the severity of the underlying histopathologic condition in HELLP syndrome. We propose that all patients with HELLP syndrome, regardless of the degree of their laboratory abnormalities, be treated aggressively, primarily with delivery.


The immunoglobulin G fraction from plasma containing antiphospholipid antibodies causes increased placental thromboxane production. Our objective was to evaluate whether the immunoglobulin G fraction from plasma containing high levels of antiphospholipid antibodies alters the production of prostacyclin or thromboxane when incubated with normal human placental tissue. The immunoglobulin G fraction was prepared from the pooled plasma of five volunteers with normal obstetric histories and no antiphospholipid antibodies. The immunoglobulin G fraction was prepared similarly from a patient with the antiphospholipid antibody syndrome. Doses of these immunoglobulin G fractions ranging from 0.3 to 3.0 mg were incubated with placental explants obtained from eight normal pregnancies, and prostacyclin and thromboxane production was assessed over 48 hours. Placental prostacyclin production was unaltered by incubation with either immunoglobulin G fraction at any of the doses tested. Placental thromboxane production tripled by 32 hours with the addition of 0.6, 1.5, and 3.0 mg of the antiphospholipid antibody fraction (p < 0.05) compared with baseline production but was unaltered by the addition of the normal pooled plasma fraction at any dose. The increase in thromboxane production with antiphospholipid antibody immunoglobulin G appeared to be dose related. The immunoglobulin G fraction prepared from plasma containing antiphospholipid antibodies caused increased placental thromboxane production without altering prostacyclin production.


The effect of magnesium sulfate on maternal retinal blood flow in preeclampsia: a randomized placebo-controlled study. Our purpose was to determine the effect of a 6 gm intravenous bolus of magnesium sulfate on maternal retinal blood flow velocity waveforms in patients with preeclampsia. Twenty-two patients were randomized to two groups and were studied with color flow Doppler before and immediately after infusion of either 6 gm of magnesium sulfate or placebo. The central retinal and posterior ciliary arteries were studied. Magnesium sulfate significantly reduced the pulsatility index (p < 0.005) in both vessels (change in pulsatility index = 0.26 +/- 0.23 for central retinal and -0.22 +/- 0.15 for posterior ciliary) when compared with placebo (change in pulsatility index = 0.02 +/- 0.10 for central retinal and -0.03 +/- 0.09 for posterior ciliary). These data suggest that magnesium sulfate dilates the vessels distal to the central retinal and posterior ciliary arteries. Changes in pulsatility index in the retinal arteries may be indicative of similar changes in other cerebral vessels.


Effect of magnesium sulfate on plasma endothelin-1 levels in normal and preeclamptic pregnancies. We attempted to determine the effects of magnesium sulfate on: (1) endothelin-1 concentration in preeclampsia, preterm labor, and term pregnancy and (2) endothelin-1 release from human umbilical cord endothelial cells. Plasma samples were prospectively collected from eight women with preeclampsia, six preterm labor patients, and eight term patients undergoing external cephalic version before and 2 hours after magnesium sulfate infusion. Supernatants were collected from human umbilical cord endothelial cells exposed to magnesium sulfate and controls. All samples were assayed with a specific radioimmunoassay for endothelin-1. Paired Student t test and analysis of variance were used for statistical analysis. Magnesium sulfate infusion in preeclampsia lowered endothelin-1 levels compared with preinfusion values (6.6 +/- 3.81 before and 4.75 +/- 2.28 after infusion, p < 0.02). Magnesium sulfate did not have an effect on endothelin-1 concentration in preterm and term pregnancies. Magnesium sulfate did not alter the endothelin-1 release from human umbilical cord endothelial cells. A significant reduction of endothelin-1 plasma levels after magnesium sulfate therapy is limited to preeclampsia. In contrast, this lowering effect was not exhibited in women without preeclampsia or in normal endothelial cells.


Correlation of nocturnal increase in plasma oxytocin with a decrease in plasma estradiol/progesterone ratio in late pregnancy. Our purpose was to determine whether plasma oxytocin concentrations show a daily rhythm. Ten women between 37 and 39 weeks of gestation volunteered for the study. They were admitted 1 to 2 days before the experiment. Three blood samples were taken with 2-minute intervals each time, at 8 AM, 4 PM, and 12 midnight. Oxytocin, 17 beta-estradiol, progesterone, and cortisol were measured by radioimmunoassay with highly specific antibodies. Statistical analysis of variance by Friedman's test was followed by multiple range testing, and p < 0.05 was considered significant. Significant daily rhythm was found for plasma cortisol, progesterone, and oxytocin and for the estradiol/progesterone ratio. Oxytocin showed a nocturnal peak and a strong negative correlation with the estradiol/progesterone ratio. The daily rhythm in plasma oxytocin parallels the rhythm in uterine activity (shown by others), suggesting a causal relationship between the two. Both may in turn be related to the ratio of circulating estradiol and progesterone.


Lack of normal increase in saliva estriol/progesterone ratio in women with labor induced at 42 weeks' gestation. Saliva specimens were collected approximately every 6 hours from day 280 to day 294 by 16 women who labored on day 294. There was a normal rise in the mean saliva estriol/progesterone ratio in the six who went into labor spontaneously but no change in the 10 whose labor was induced.


HLA-DQ beta 57 in Hispanic patients with insulin-dependent diabetes mellitus. The purpose of our study was to investigate the distribution of HLA-DQ beta-chain amino acid residue 57 (HLA-DQ beta 57) as a genetic marker of susceptibility for insulin-dependent diabetes mellitus in the Hispanic population. Fifteen patients of Puerto Rican descent with juvenile-onset insulin-dependent diabetes mellitus underwent human leukocyte antigen typing for HLA-DQ beta 57 by polymerase chain reaction amplification of the target genomic DQ sequence followed by hybridization of the polymerase chain reaction product to phosphorus 32-labeled allele-specific oligonucleotide probes. A control group of 44 Hispanic adults without diabetes who were undergoing human leukocyte antigen typing for tissue donation were concurrently typed for comparison. The Hispanic insulin-dependent diabetes mellitus group showed a significant increase in homozygosity for a non-aspartate amino acid (p = 0.023) over a control group of Hispanic subjects without diabetes. A high rate of heterozygosity for aspartate (53.3%) is found in Hispanic subjects with insulin-dependent diabetes mellitus as well. HLA-DQ beta 57 in the Hispanic population has a distribution distinct from HLA-DQ beta 57 in the Caucasian population. A single aspartate is not protective against insulin-dependent diabetes mellitus in Hispanic subjects.


Changes in T-lymphocyte subpopulations during pregnancy complicated by human immunodeficiency virus infection. We describe changes in T-lymphocyte subpopulations in pregnancies complicated by human immunodeficiency virus infection. T-lymphocyte counts were performed every trimester and post partum on all patients with human immunodeficiency virus infection. Patients with a CD4 count < 500 cells/mm3 were offered zidovudine after the first trimester. We performed a regression analysis of repeated measures on the total lymphocyte, CD4, and CD8 counts. Twenty-three patients were studied, 10 (43%) of whom took zidovudine. CD4, CD8, and total lymphocyte counts decreased in patients not taking zidovudine but remained stable in patients receiving zidovudine. The differences were not statistically significant. CD4 and CD8 counts may decrease during pregnancies complicated by human immunodeficiency virus infection, primarily because of decreases in the total lymphocyte count. The use of zidovudine may prevent this decline. The clinical use of absolute CD4 counts during pregnancy requires further study.


Use of esterase inhibitors and zone electrophoresis to define bacterial esterases in amniotic fluid. The purpose of our study was to define further the role of bacterial esterases in amniotic fluid obtained from women with chorioamnionitis. Amniotic fluid samples from 39 patients with chorioamnionitis were submitted for bacterial cultures and in vitro assay. Esterase inhibitors diisopropyl fluorophosphate and iodoacetic acid were added and the degree of inhibition calculated. These results were compared with the amniotic fluid culture results. Chi square analysis was performed to compare the results of the esterase assay and the inhibition assay between the uninfected and infected amniotic fluid samples. Thirty-one patients had positive bacterial cultures, with 21 being infected with gram-negative organisms. All samples showed significant inhibition (range 55% to 82%) with diisopropyl fluorophosphate. There was partial inhibition with iodoacetic acid (range 10% to 30%) in the gram-negative samples but no inhibition in the gram-positive and uninfected samples. Six infected and two uninfected samples were analyzed by using zone electrophoresis with human plasma as a control. Minimal esterase motility was noted in the amniotic fluid samples as compared with that in plasma. The esterases in amniotic fluid appeared to be of bacterial, not human, origin. Furthermore, different groups of bacteria appeared to produce different esterases in infected amniotic fluid.


Amniotic fluid tumor necrosis factor-alpha and interleukin-1 in a rabbit model of bacterially induced preterm pregnancy loss. The objective of this study was to determine whether the cytokines tumor necrosis factor-alpha, interleukin-1 alpha, and interleukin-1 beta were produced in the amniotic fluid of the rabbit after intracervical inoculation with Escherichia coli. Timed pregnant rabbits on day 21 (70% of gestation) were inoculated with a hysteroscope intracervically with 10(4) to 10(5) colony-forming units Escherichia coli or sterile saline solution. Escherichia coli-inoculated animals (N = 16) were put to death at 4, 8, 12, and 16 hours after inoculation. Control animals (N = 6) were put to death at similar intervals. At death, cultures were taken from endometrium, amniotic fluid, peritoneum, and blood. Amniotic fluid was collected and assayed for tumor necrosis factor bioactivity by a modified fibroblast cytotoxic assay in L929 cells, for interleukin-1 alpha, and interleukin-1 beta with a specific radioimmunoassay, and for prostaglandin E2 and prostaglandin F2 alpha by radioimmunoassay. Levels of amniotic fluid tumor necrosis factor-alpha, interleukin-1 alpha, and interleukin-1 beta were elevated as early as 4 hours after inoculation in some animals and by 12 to 16 hours after inoculation in all. Levels of all three cytokines correlated significantly with time from intracervical inoculation with Escherichia coli (p < 0.05). Levels of amniotic fluid prostaglandin E2 and prostaglandin F2 alpha correlated significantly with time from intracervical inoculation with Escherichia coli (p < 0.05). Levels of tumor necrosis factor-alpha, interleukin-1 alpha, interleukin-1 beta, prostaglandin E2 and prostaglandin F2 alpha are elevated in the amniotic fluid of rabbits after intracervical inoculation with Escherichia coli. Similarity exists between elevations of amniotic fluid cytokines in this model and in cases of intraamniotic infection and preterm labor unresponsive to tocolytics in humans. Modulation of cytokines may offer a strategy for improvement of outcome in this experimental model of infection-induced pregnancy loss.


Association of cervicovaginal infections with increased vaginal fluid phospholipase A2 activity. The purpose of this study was to determine if phospholipase A2 was detectable within vaginal fluid and to correlate its presence with the presence of common lower genital tract infection or microbial conditions. Pregnant women were examined at the first prenatal visit with standard clinical evaluations and microbiologic cultures or tests. Vaginal fluid samples were evaluated for phospholipase A2 activity by means of a standardized enzyme fluorometric assay. Data were stratified to control for coexisting infections. Phospholipase A2 activity was detected among 29.8% of women and was independently associated with the presence of bacterial vaginosis (p < 0.001), Trichomonas. vaginalis (p < 0.04), and Chlamydia trachomatis (p < 0.02). The percentage of women with phospholipase A2 activity and the level of activity was increased in the presence of more than one infection. Elevated reproductive tract phospholipase A2 concentrations among pregnant women may play roles in the pathogenesis of preterm labor and birth. Identification of pregnant women with increased concentrations in vaginal fluid may allow for development of effective intervention strategies to reduce the risk of preterm birth.


Transfer of Timentin (ticarcillin and clavulanic acid) across the in vitro perfused human placenta: comparison with other agents. In vitro perfusion of human placentas was used to quantify the net placental transfer of ticarcillin and clavulanic acid. Placentas were obtained from uncomplicated pregnancies at term. The maternal and fetal circulations were reestablished at flow rates of 17.5 ml/min and 5 ml/min, respectively. Open circulations were used to evaluate steady-state pharmacodynamics and transplacental gradient formation. Drug levels were measured by high-pressure liquid chromatography. The clearance index of ticarcillin was 0.037 +/- 0.004. The fetal/maternal ratio was 0.91. Therapeutic concentrations of clavulanate (2 to 6 micrograms/ml) in the maternal media resulted in undetectable transfer to the fetal compartment. By using higher levels of clavulanate, a clearance index of 0.061 +/- 0.001 (mean +/- SEM) and 1:1 fetal/maternal gradient was obtained. These data correspond to relatively low transfer of ticarcillin with a cord/maternal ratio of < 1. Clavulanate transfer is slightly greater. Agents with similar activity and superior transfer would optimize intrauterine treatment.


Nitric oxide and prostacyclin inhibit fetal platelet aggregation: a response similar to that observed in adults. We evaluated the relative importance of two endothelium-derived substances, prostacyclin and nitric oxide, in their ability to inhibit aggregation of fetal and maternal platelets. The effects of various concentrations of prostacyclin and S-nitroso-N-acetylpenicillamine (which releases nitric oxide) on platelet aggregation were studied by means of platelet-rich plasma from at least five to six subjects per group. Fetal blood was collected from umbilical vein at delivery. Maternal venous blood was collected within 4 hours of delivery. Platelet aggregation was monitored with a platelet aggregation profiler. Adenosine diphosphate was used as the aggregating agent. Statistical differences between means were evaluated with two-way analysis of variance or Student t test. Prostacyclin and S-nitroso-N-acetylpenicillamine inhibited aggregation of fetal and maternal platelets, but prostacyclin was more potent. Fetal platelets were more sensitive than maternal platelets to prostacyclin and S-nitroso-N-acetylpenicillamine. Prostacyclin appears to be more important in preventing aggregation of platelets in the feto placental circulation.


Peripheral magnesium sulfate enters the brain and increases the threshold for hippocampal seizures in rats. Our objectives were to determine whether magnesium sulfate crosses the blood-brain barrier and whether it has central anticonvulsant action. In experiment 1 34 female Long-Evans rats were divided into six groups: control (n = 7); single magnesium sulfate injection and evaluation after 20 minutes in 3 conditions: normal rats (n = 7), sham-operated animals (n = 5), and after electrical stimulation by hippocampal electrode (n = 5); single injection and evaluation after 2 hours (n = 5); and prolonged (2 hours) serum magnesium elevation (n = 5). Serum, cerebrospinal fluid, and specific brain areas were analyzed for magnesium concentrations. In experiment 2 threshold for electrical seizure was measured in eight rats before and after intraperitoneal injections of magnesium sulfate versus saline solution. In experiment 1 there was a significant correlation between blood and cerebrospinal fluid magnesium concentrations (r = 0.80, p < 0.0001). Magnesium concentrations were increased in the cortex and hippocampus, with the largest changes occurring after two hours of sustained serum magnesium concentrations (p < 0.01). Induction of hippocampal seizure activity resulted in further elevations in cerebrospinal fluid magnesium concentrations but did not change brain concentrations. In experiment 2 magnesium sulfate increased the electrical threshold required to induce seizures by 34% (p = 0.01). Magnesium sulfate enters the cerebrospinal fluid and brain and has a central anticonvulsant effect.


Prenatal exposure to phenobarbital and quantifiable alterations in the electroencephalogram of adult rat offspring. Our purpose was to measure central nervous system insult from prenatal phenobarbital by quantified electroencephalographic evaluation of mature rat offspring. Twenty-four adult female rats were given phenobarbital as 0, 20, 40, or 60 mg/kg/day subcutaneously between 9 and 10 AM for 28 days before breeding and throughout gestation. The electroencephalograms of 90-day-old offspring were sampled over 24 hours and converted to the power spectra. Female offspring exposed prenatally to 20 mg and males exposed prenatally to 40 mg displayed an increase in delta and a decrease in alpha and middle-beta waveform activities. Females exposed prenatally to 40 mg and males exposed prenatally to 60 mg displayed decreases in delta, theta, alpha, and middle-beta activities. Prenatal phenobarbital exposure produced dose- and sex-dependent changes in the electroencephalogram of mature offspring consistent with a generalized deficit in the neuronal synchronization mechanisms that are critical for normal sleep and learning.


Fetal catecholamine, cardiovascular, and neurobehavioral responses to cocaine. Although maternal cocaine administration results in fetal cardiovascular and behavioral alterations, these responses have been attributed to hypoxia resulting from reduced uteroplacental blood flow. We studied the fetal catecholamine, cardiovascular, and neurobehavioral responses to direct fetal cocaine administration. Cardiovascular, electrocortical, and electroocular responses and plasma catecholamines were monitored in chronically catheterized fetal sheep (n = 7), 127 +/- 2 days' gestation, during a control period and after intravenous fetal injections of cocaine 0.5 and 1.0 mg/kg. Intravenous cocaine caused prompt increases in fetal plasma norepinephrine (372 +/- 73 to 531 +/- 112 and 842 +/- 233 pg/ml), epinephrine (27 +/- 7 to 46 +/- 13 and 49 +/- 11 pg/ml), and systolic blood pressure (46 +/- 2 to 53 +/- 2 and 55 +/- 2 mm Hg). Low-voltage electrocortical activity decreased from 61.7% +/- 3.0% to 38.4% +/- 3.9% in the first hour after the cocaine 0.5 mg/kg dose but recovered to baseline values during the second hour. After the cocaine 1.0 mg/kg dose, low-voltage electrocortical activity decreased to 40.7% +/- 2.0% and did not recover thereafter. Fetal blood gas values did not change. Direct fetal cocaine administration increases fetal plasma catecholamine levels and fetal blood pressure and suppresses low-voltage electrocortical activity. Chronic cocaine exposure may hamper central nervous system maturation and alter postnatal development.


Fetal hemodynamic response to maternal intravenous nicotine administration. Our study was designed to test the hypothesis that maternally administered nicotine has significant effects on fetal hemodynamics and umbilical systolic/diastolic ratios. Nine pregnant ewes received maternal intravenous infusions of 10, 20, and 30 micrograms/kg/min of nicotine. Maternal and fetal blood pressure, heart rate, and uterine and umbilical blood flow were recorded. Maternal intravenous administration of nicotine (10, 20, and 30 micrograms/kg/min of maternal body weight) produced significant (p < 0.05) increases in fetal blood pressure (2%, 11%, and 25%, respectively), decreases in fetal heart rate (0%, 8%, and 12%), and decreases in umbilical blood flow (0%, 0%, and 19%). Umbilical systolic/diastolic ratios increased slightly at the 30 micrograms/kg/min dose of nicotine, but these changes did not reach significance. Maternal blood pressure increased (10%, 25%, and 53%), and uterine vascular resistance increased (5%, 64%, and 344%) significantly (p < 0.05); uterine blood flow increased at the 10 micrograms/kg/min dose (+5%) and decreased by 23% and 42% at the highest two doses of nicotine. Maternal nicotine administration in late-term pregnant sheep produced significant increases in fetal arterial blood pressure and umbilical vascular resistance, decreased fetal heart rate, and umbilical blood flow but did not significantly alter systolic/diastolic ratios.


The concentration of estrogen receptors in rabbit uterine myocytes decreases in culture. The purpose of this study was to determine whether the concentration of estrogen receptors in cultured myocytes is preserved after dispersion. Primary myocytes were prepared from rabbit myometrium by collagenase dispersion after removing the endometrium and were isolated with Percoll density gradients. The cells were assayed for estrogen receptor concentration at intervals after dispersion by means of a whole-cell binding assay. Unpaired t test was used for comparisons. The concentration of estrogen receptors on the first day after dispersion was 12,058 +/- 1096 sites per cell (mean +/- SEM) and decreased to 4389 +/- 1223 site per cell within 9 to 14 days after dispersion (63% decline, p < 0.001). A similar decrease was observed when 2 nmol/L estradiol was present in the medium. The concentration of estrogen receptors in isolated rabbit uterine myocytes decreases after dispersion. This may partly explain the difficulty of demonstrating in vitro estrogen effects on myocytes, which are well established in vivo.


Myometrial contractile responsiveness to oxytocin after dexamethasone suppression of circadian uterine activity in pregnant rhesus macaques during late gestation. This study was designed to determine if dexamethasone alters myometrial responsiveness to oxytocin or oxytocin secretion. Studies were conducted in rhesus macaques (n = 6), between 144 and 148 days' gestation (term 167 days). The first study was conducted at 9 AM and repeated 36 hours later at 9 PM. At 9 AM the following morning a continuous maternal dexamethasone infusion (15 micrograms/kg/hr given intravenously) was initiated, and the study was repeated at 9 PM, 60 hours later. Four doses of oxytocin (500, 1000, 2000, and 4000 pg/kg/min) were administered as 1-minute pulses every 5 minutes for 30 minutes. The number of contractions per pulse (contraction/pulse ratio) was used to determine differences in myometrial responsiveness. Before dexamethasone infusion there was a circadian rhythm in uterine activity with peak contractile events between 8 and 10 PM (p < 0.01), whereas during infusion the rhythm was ablated. At oxytocin dose 1, the 9 AM contraction/pulse ratio (0.3 +/- 0.1) was lower than that for 9 PM (0.6 +/- 0.2) and for 60 hours later (0.6 +/- 0.1) (mean +/- SE, p < 0.05). Similar results were observed at dose 2, whereas no differences in the contraction/pulse ratio were noted at dose 3. Basal plasma oxytocin concentrations were unaffected by dexamethasone treatment, whereas plasma estradiol and cortisol concentrations were reduced compared with control values (p < 0.01). (1) There is a differential sensitivity to oxytocin between morning and evening and (2) the dexamethasone-induced loss of the uterine contractile rhythm is not the result of a loss of myometrial sensitivity to oxytocin or to a suppression of plasma oxytocin concentrations.


Effect of endothelin-1 on the uterine vasculature of the pregnant and estrogen-treated nonpregnant sheep. This study was designed to evaluate the uterine vascular responses to endothelin-1 in pregnant and estrogen-treated nonpregnant sheep. Seven pregnant and five nonpregnant oophorectomized ewes received local uterine artery infusions of endothelin-1, norepinephrine, and phenylephrine. Arterial blood pressure, heart rate, and uterine blood flow were recorded. Endothelin-1 (0.01 to 3.0 micrograms/min), norepinephrine (0.1 to 3 micrograms/min), and phenylephrine (0.1 to 10 micrograms) produced significant dose-related decreases in uterine blood flow and increases in uterine vascular resistance. On a nanomoles infused per minute basis, endothelin-1 was much more potent than norepinephrine and phenylephrine as a uterine artery vasoconstrictor in both pregnant and nonpregnant sheep. The uterine vascular responses to norepinephrine and phenylephrine were similar in pregnant and nonpregnant ewes, whereas response to endothelin-1 was blunted in pregnancy. Endothelin-1 is an extremely potent uterine vasoconstrictor in both pregnant and nonpregnant ewes, but the uterine vascular responsiveness to endothelin-1 is decreased in pregnancy.


The comparative effects of big endothelin-1, endothelin-1, and endothelin-3 in the human fetal-placental circulation. This study compared the effects of big endothelin-1, endothelin-1, and endothelin-3 and whether endothelin-converting enzyme was present in the human fetal-placental circulation. Single cotyledons of term placentas were dually perfused in vitro, and increases in fetal-placental perfusion pressure to bolus injections of big endothelin-1, endothelin-1, and endothelin-3 (8 x 10(-10) to 1 x 10(-7) mol/L) were recorded. Responses to big endothelin-1 (10(-7) mol/L) were measured in the same placenta before and after perfusion of the fetal-placental circulation with the neutral metalloprotease inhibitor phosphoramidon (10(-5) mol/L), which acts as an endothelin-converting enzyme inhibitor. All experiments were performed in at least five separate placentas. Significant concentration-dependent increases in fetal-placental perfusion pressure were seen with endothelin-1 (p < 0.0005), endothelin-3 (p < 0.0256), and big endothelin-1 (p < 0.0034, analysis of variance). Big endothelin-1 always elicited transient vasodilatation before constriction. Phosphoramidon significantly inhibited the vasoconstrictor effect of big endothelin-1 (p < 0.039, paired t test). The three endothelins tested are vasoconstrictors, and endothelin-converting enzyme is present in the fetal-placental circulation.


Decrease in annexin I messenger ribonucleic acid expression in human amnion with labor. Annexins are a superfamily of proteins that are thought to inhibit phospholipase A2 activity and hence inhibit prostaglandin production. The purpose of this study was to test the hypothesis that annexin I concentration in human amnion is reduced with labor and that this reduction is mediated by a decrease in annexin I messenger ribonucleic acid expression. Amnion and choriodecidua were collected from term singleton pregnancies, eight after spontaneous vaginal delivery and eight from elective cesarean section without labor. Annexin I protein was quantitated by Western blotting. Ribonucleic acid was isolated from amnion, and then annexin I messenger ribonucleic acid was identified by Northern hybridization and quantitated by slot blotting. Annexin I (35 kd) was identified in amnion tissue. The concentration in the group undergoing labor (320 +/- 45 integrated optical density units, mean +/- SE) was significantly reduced (p < 0.05) compared with that in the group not undergoing labor (635 +/- 65 units). The size of the annexin I messenger ribonucleic acid was approximately 1.8 kb. The mean integrated optical density for the labor group (840 +/- 139 units, mean +/- SE) was significantly reduced (p < 0.05) compared with that of the nonlabor group (1912 +/- 464 units). There is a significant decrease in annexin I messenger ribonucleic acid expression in human amnion with labor, corresponding to a significant decrease in annexin I protein concentration. This may contribute to the increased phospholipase A2 activity, arachidonic acid mobilization, and prostaglandin production at labor in humans.


Validation of noninvasive fetal renal artery flow measurement by pulsed Doppler in the lamb. Our objective was to evaluate the accuracy of quantitative measurement of blood flow of the left renal artery in the fetal lamb by means of range-gated, two-dimensional pulsed Doppler ultrasonography. Doppler measurements were compared with invasive measurements obtained with a perivascular flow probe (Transonic) placed on the fetal renal artery. Renal blood flow was manipulated acutely either by volume expansion or depletion or by means of pharmacologic agents. With each manipulation values of the left fetal renal artery blood flow were obtained with both methods. The size of the fetal renal artery was determined by postmortem examination. A total of 36 paired flow measurements were obtained on five fetuses. One fetus had only Transonic determination of renal blood flow. Pulsed Doppler ultrasonography blood flow and transit-time ultrasonography blood flow estimations were correlated (F = 82.4, R2 = 0.73, p < 0.001). This study suggests that two-dimensional pulsed Doppler can reliably estimate renal blood flow in the fetal lamb.


Effects of maternal administration of 3% carbon dioxide on umbilical artery and fetal renal and middle cerebral artery Doppler waveforms. The null hypothesis is that umbilical, middle cerebral, and renal artery pulsed Doppler velocity waveforms in the normal term fetus may be affected during short-term maternal inhalation of 3% carbon dioxide gas mixture. Seventy-two observations were made on 14 term fetuses before and during maternal 3% carbon dioxide gas mixture inhalation. The umbilical, middle cerebral, and renal arteries of these fetuses were sampled with pulsed Doppler velocity waveforms and recorded on a strip chart at a preset speed of 50 mm/sec. Doppler waveforms were analyzed for differences in the systolic peak to end-diastolic velocity ratio for these three vascular beds. Peak flow velocity and time velocity integral were also analyzed for the cerebral and renal vascular beds. The data were analyzed with the paired t test. A significant decrease in the systolic-to-diastolic-velocity ratio of the middle cerebral artery occurred with 3% carbon dioxide inhalation (p < 0.02). The other vascular beds had no demonstrable change. Transient maternal breathing of 3% carbon dioxide gas mixture selectively causes a decrease in resistance in the fetal cerebral circulation.


Differential effects of intravenous hydralazine on myoendometrial and placental blood flow in hypertensive pregnant ewes. The differential vasoactive effects of hydralazine on the uteroplacental vascular bed were studied. After control measurements were taken, near-term chronically prepared pregnant sheep were continuously infused with angiotensin II. Maternal arterial pressure was increased by 32 mm Hg. Hydralazine was then administered; the effects on regional resistance and blood flow were evaluated with a radionuclide-labeled microsphere technique. Analysis of variance for repeated measures was used to compare observations. When compared with the hypertensive state, hydralazine caused the following changes by 40 minutes (mean +/- SEM): Although maternal blood pressure fell 31% +/- 5% (p = 0.0005), placental blood flow was unchanged, total uteroplacental blood flow increased 24% +/- 8% (p = 0.03), total uteroplacental resistance decreased 43% +/- 4% (p = 0.0002), placental resistance decreased 19% +/- 9% (p = 0.01), myoendometrial blood flow increased 390% +/- 82% (p = 0.0005), and myoendometrial resistance decreased 82% +/- 4% (p = 0.0005). In angiotensin II-induced hypertensive ewes, hydralazine is an effective dilator of the uteroplacental vascular bed and can maintain placental blood flow while blood pressure.


Endogenous modulation of the blunted adrenergic response in resistance-sized mesenteric arteries from the pregnant rat. We tested the hypothesis that during pregnancy the endothelium mediates the blunted response to adrenergic vasoconstriction. Mesenteric resistance arteries from late pregnant (n = 6) and age-matched virgin control (n = 6) Sprague-Dawley rats were studied in a myograph. Arteries from pregnant rats were 35% less sensitive to phenylephrine vasoconstriction than were those from nonpregnant rats (mean effective concentration that produced a 50% response 2.26 vs 1.48 mumol/L, pregnant vs nonpregnant, p < 0.01). Meclofenamate had no effect on the vasoconstrictor response in arteries from either group. Inhibition of endothelium-derived relaxing factor with N(o)-nitro-L-arginine methyl ester or endothelial cell removal had a similar twofold increase in phenylephrine sensitivity in arteries from both the pregnant and nonpregnant rats (mean effective concentration that produced a 50% response 2.26 vs 1.11 mumol/L for pregnant rats and 1.48 vs 0.72 mumol/L for nonpregnant rats, p < 0.01). However, methacholine relaxation response was potentiated in pregnant versus nonpregnant rats (mean effective concentration that produced a 50% response 0.030 vs 0.049 mumol/L, p < 0.01). Although the potential for endothelium-dependent relaxation is augmented in mesenteric arteries of the pregnant rat, the decreased sensitivity to phenylephrine during pregnancy is not modulated acutely by endothelium-derived relaxing factor or by prostaglandin products of the cyclooxygenase pathway.


A unique hypertonic response to hypotonic infusion in the pregnant ewe. The purpose of this study was to compare the responses of the maternal ewe to intravenous volume expansion with either sufficient lactated Ringer's solution to elevate maternal venous pressure or sufficient hypotonic fluid to reduce blood osmolality. Chronically catheterized pregnant sheep were intravenously infused over 4 hours with either commercial lactated Ringer's solution (5.55 +/- 0.50 L/hr, 255 mOsm/kg, mildly hypotonic) or diluted Ringer's solution (2.04 +/- 0.27 L/hr, 150 mOsm/kg, markedly hypotonic). Data were statistically analyzed with two- and three-factor analyses of variance and bivariate regression analysis. During the mildly hypotonic infusion (n = 8) the maternal blood osmolality changes were -5.1 +/- 1.2, +2.7 +/- 1.0 and +6.8 +/- 1.1 mOsm/kg at 1 and 4 hours of infusion and 1 hour after the infusion. In four of the eight animals in this group profuse diarrhea developed. During the markedly hypotonic infusion (n = 11) the maternal blood osmolality changes were -9.9 +/- 1.1, -15.9 +/- 2.5, and -10.4 +/- 2.2 mOsm/kg at 1 and 4 hours of infusion and 1 hour after the infusion. Although urine osmolalities were significantly less than the osmolality of the infusate in both groups, only during the mildly hypotonic infusion was there a net loss of free water by the kidneys. The renal free water loss, the venous pressure increase, and the blood osmolality decrease were not significantly different whether diarrhea did or did not develop. The infusion of large volumes of mildly hypotonic Ringer's solution to the pregnant ewe produces a paradoxic increase in maternal plasma osmolality as a result of the excretion of large volumes of free water by the kidneys, and if the venous pressure is increased more than about 6 mm Hg with this infusion, diarrhea develops in the animals.


Dehydration increases the renal response to atrial natriuretic peptide in fetal sheep. In sheep, maternal water deprivation results in urinary natriuresis in spite of suppression of plasma atrial natriuretic factor levels. Near-term fetal sheep also have a urinary natriuresis without change in plasma atrial natriuretic factor during maternal dehydration. This study was designed to explore the role of plasma atrial natriuretic factor levels in fetal dehydration-natriuresis. Eight chronically instrumented preterm (113 +/- 1 days) ovine fetuses received two atrial natriuretic factor infusions (3 and 15 ng/kg/min) in a euhydrated state and after 48 +/- 1 hours of maternal water deprivation. Dehydration significantly increased maternal plasma osmolality (302 +/- 2 to 313 +/- 2 mOsm/kg water), sodium (148.1 +/- 0.8 to 154.3 +/- 0.4 mEq/L), chloride (112.4 +/- 0.6 to 116.8 +/- 0.9 mEq/L), and arginine vasopressin (4.2 +/- 1.2 to 23.0 +/- 4.0 pg/ml) and significantly decreased plasma atrial natriuretic factor (36 +/- 6 to 19 +/- 4 pg/ml) concentrations. Fetal plasma osmolality (296 +/- 1 to 308 +/- 2 mOsm/kg), atrial natriuretic factor (128 +/- 16 to 241 +/- 36 pg/ml), and arginine vasopressin (3.5 +/- 0.8 to 12.3 +/- 4.8 pg/ml) concentrations and urine osmolality (170 +/- 10 to 253 +/- 10 mOsm/kg), osmolar clearance (0.80 +/- 0.02 to 0.14 +/- 0.02 ml/kg/min), and fractional sodium excretion (3.3% +/- 1.7% to 8.5% +/- 2.1%) increased significantly with dehydration, whereas the plasma atrial natriuretic factor clearance decreased from 127 +/- 27 to 63 +/- 10 ml/kg/min. Dehydration had no effect on fetal hematocrit, vascular pressures, glomerular filtration rate, urine flow, or free water clearance. In euhydrated fetuses plasma atrial natriuretic factor increased from 128 +/- 16 to 287 +/- 46 pg/ml with sequential atrial natriuretic factor infusion, and no significant increases were observed in urine flow, fractional sodium excretion, and glomerular filtration rate. In contrast, atrial natriuretic factor infusion to dehydrated fetuses significantly increased urine flow (0.17 +/- 0.03 to 0.32 +/- 0.07 ml/kg/min), osmolar clearance (0.14 +/- 0.02 to 0.28 +/- 0.06 ml/kg/min), and fractional sodium excretion (8.5% +/- 2.1% to 14.8% +/- 4.0%). These results demonstrate that in the fetus at 113 days' gestation plasma atrial natriuretic factor levels increase with dehydration, probably a result of decreased plasma atrial natriuretic factor clearance, and the fetal renal responsiveness to atrial natriuretic factor infusion increases during maternal dehydration.


Ovine maternal and fetal renal vasopressin receptor response to maternal dehydration. Arginine vasopressin secretion increases in response to increased plasma osmolality or hypovolemia. Dehydration-induced increases in plasma arginine vasopressin levels have been shown to down-regulate arginine vasopressin V2 receptors in adult rat kidneys. Our study determined ovine maternal and fetal renal arginine vasopressin receptor characteristics and receptor response to maternal dehydration. Eight pregnant ewes (113 +/- 1 days) were dehydrated for 72 hours; eight animals served as controls. Renal medullary tissue was isolated from maternal and fetal kidneys, and arginine vasopressin receptor characteristics determined with saturation and competition assays using tritiated arginine vasopressin, arginine vasopressin, and arginine vasopressin analogs. Euhydrated maternal and fetal renal medullary arginine vasopressin receptor dissociation constant (3.0 +/- 0.3 and 1.9 +/- 0.3 nmol/L) and maximal binding capacity (149 +/- 15 and 111 +/- 33 fmol/mg protein) values were similar. Pharmacologic profiles with selective agonists indicated a predominance of V2 receptors. Dehydration significantly increased maternal and fetal plasma osmolalities (304 +/- 2 to 320 +/- 2; 296 +/- 1 to 319 +/- 3 mOsm/kg water, respectively) and arginine vasopressin levels (3.8 +/- 1.4 to 29.3 +/- 4.6; 4.4 +/- 1.0 to 16.9 +/- 5.0 pg/ml, respectively) but had no effect on arginine vasopressin receptor binding. Specific, saturable, single-site tritiated arginine vasopressin binding is present in ovine maternal and fetal renal medullary membranes. Ovine maternal and fetal renal arginine vasopressin receptors do not down-regulate in response to dehydration-induced elevations in plasma arginine vasopressin levels.


Indomethacin-induced urinary flow rate reduction in the ovine fetus is associated with reduced free water clearance and elevated plasma arginine vasopressin levels. The purpose of our study was to explore the urinary responses of the ovine fetus to indomethacin levels comparable with those used therapeutically in the human fetus. After a 1-hour control period, chronically catheterized ovine fetuses between 125 and 139 days of gestation were given an intravenous bolus of indomethacin (0.05 mg/kg estimated fetal weight) followed by a 0.0025 mg/kg/min continuous infusion for 5 hours. The experimental group (n = 9) was compared with a vehicle-only infusion group (n = 10). There was a sustained 55.7% +/- 9.5% (mean +/- SEM) decrease in urinary output by 2 hours of indomethacin infusion (p < 0.00001, analysis of variance). Urinary osmolality, potassium, and chloride concentrations underwent sustained increases during the infusion period (p < 0.005). Free water clearance decreased by 67.5% +/- 12.0% (p < 0.001). Fetal arterial pressure increased only transiently (p < 0.05), and increases in venous pressure (p = 0.013) and heart rate (p < 0.0001) were sustained. Fetal plasma arginine vasopressin concentration increased during indomethacin infusion (p < 0.05) and was correlated with the fall in urinary flow rate and free water clearance (p = 0.002). During vehicle infusion no significant changes were observed in any of the variables. Our data indicate that the fetus undergoes antidiuresis when exposed to low levels of indomethacin and that the observed antidiuresis is mediated by a decrease in free water clearance. The reduction in free water clearance may be mediated by increases in plasma arginine vasopressin concentrations.


Amniotic fluid composition changes during urine drainage and tracheoesophageal occlusion in fetal sheep. Recently an intramembranous pathway was reported in the ovine fetus as a route for the rapid exchange of water, ions, and molecules between the amniotic fluid and the fetal blood that perfuses the fetal surface of the placenta and the fetal membranes. Our study was designed to test the hypothesis that the amniotic fluid composition would gradually equilibrate with fetal plasma when the major flows to and from the amniotic compartment were eliminated. Eleven near-term fetal sheep underwent ligation of the urachus to eliminate the allantoic fluid. An inflatable cuff was placed around the esophagus and trachea, and catheters were placed in the fetal urinary bladder, fetal circulation, and maternal circulation. At > or = 5 days after surgery the animals were subjected to either a control experiment or a continuous urine drainage plus tracheoesophageal occlusion for 8 hours. During the urine drainage plus occlusion study, amniotic fluid osmolality (p < 0.0001), Na+ (p < 0.0001), K+ (p < 0.01) Cl- (p < 0.001), and lactate (p < 0.001) increased compared with the control experiment. These corresponded to 50% reductions in the gradients for osmolality and Na+ between fetal plasma and amniotic fluid; the K+ gradient increased, and the Cl- gradient reversed. The percentage increases in amniotic Na+, K+, Cl-, and lactate were all 10% at 8 hours. These observations suggest that water is absorbed from the amniotic fluid through the intramembranous pathway into the fetal circulation at a rate of 1.25% of the total amniotic volume per hour or approximately 240 ml/day.


Rapid intramembranous absorption of water infused into the ovine allantoic cavity. Previously we found that water infused into the ovine amniotic cavity was rapidly absorbed into the fetal circulation through the vascularized fetal membranes and fetal surface of the placenta (i.e., the intramembranous pathway). The purposes of this study were to (1) estimate the conductance of the intramembranous pathway from the allantoic cavity and (2) determine if the conductance is adequate to offset the inflow of urine, which may be up to 500 ml/day in the near-term ovine fetus. Seven chronically catheterized fetal sheep averaging 132 +/- 2 (+/- SE) days' gestation underwent an infusion of warmed distilled water into the allantoic cavity at 6 ml/min. The infusions were continued until steady states were obtained in allantoic and amniotic fluid and in fetal and maternal blood osmolalities. During the steady state the conductance of the intramembranous pathway was estimated as the ratio of osmotic gradient to infusion rate. The allantoic and amniotic fluid and the fetal and maternal blood osmolalities decreased by 188 +/- 14, 36 +/- 8, 13 +/- 2, and 3 +/- 1 mOsm/kg, respectively, at steady state. From the fetal-allantoic osmolality gradients the conductance of the intramembranous pathway was 1.72 +/- 0.14 or 0.53 +/- 0.08 microliter/min/mm Hg/kg fetal weight. Assuming a similar conductance during the preinfusion period, the next volume movement would equal 0.67 ml/min (965 ml/day). The conductance of the intramembranous pathway in combination with the normal osmotic gradient is sufficient to remove the large volume of fetal urine that may enter the allantoic cavity each day.


Vascularization of the ovine amnion and chorion: a morphometric characterization of the surface area of the intramembranous pathway. The purpose of this study was to characterize the vascularization of the ovine amnion, allantois, and chorion. A white silicone vascular casting material was infused into both umbilical arteries of nine fetal sheep ranging in age from 58 to 142 days' gestation. A morphometric analysis of photomicrographs of the membranes was then performed with computerized image analysis techniques. After removal of the uterus, the fetus was surrounded by a layer of white silicone-filled microvessels in the chorion. The amniotic membrane after separation from the chorion was covered by a fine mesh of microvessels, whereas the allantois was avascular. The amniotic membrane readily separated into an outer vascularized layer and an inner, avascular layer containing the amnion. Approximately 50% of the surface of the chorionic membrane was covered by microvessels; this appeared independent of gestational age. At midgestation 30% of the surface of the amnion was covered by microvessels, and this decreased to 17% at 142 days. Relative to fetal weight, the amniotic and chorionic vascular surface areas decreased from 6 to 0.3 and from 15 to 1.5 cm2/gm fetal weight, respectively, over the last half of gestation. There is an extensive network of microscopic fetal blood vessels within the ovine chorion and covering the outer surface of the amnion. These vessels appear to be ideally situated to facilitate a direct exchange of water and solutes between amniotic or allantoic fluids and fetal blood through the intramembranous pathway.


Increased lymphatic pumping ability in pregnant ewes. Daily the lymphatic system returns several liters of fluid to the systemic circulation. Unlike blood, lymph is pumped against a pressure gradient. Because vessels in the systemic circulation are hyporesponsive to vasoconstrictor hormones during pregnancy, we decided to assess whether lymphatic pumping ability was decreased during pregnancy. Ten pregnant ewes were chronically catheterized in the left thoracic duct and jugular vein. Four days after surgery thoracic duct pumping ability was determined by measuring the lymph flow rate as a function of outflow pressure by varying the height at which lymph flow drains in an open-flow system. Lymph flow was plotted against outflow pressure, and the value at which lymph flow decreases is defined as the "breaking point." The outflow pressure at which lymph flow equals zero is the "stopflow" pressure; this value defines the pumping ability. The pregnant ewes had a breaking point of -1 +/- 2.7 versus 14.4 +/- 3.0 mm Hg (mean +/- SD) for the nonpregnant sheep. Stopflow pressures were 62 +/- 7.3 versus 23.9 +/- 4.7 mm Hg (mean +/- SD) for the nonpregnant ewes (p < 0.05). The ability of the lymphatic system to generate pressure is increased during pregnancy, but the breaking point is diminished. Alterations of lymphatic pumping ability could explain fluid retention under pathophysiologic conditions.


Prediction of fertilization in vitro with human gametes: is there a litmus test? The objectives of this study were as follows: (1) to evaluate the relationships between sperm concentration, morphologic pattern, motion parameters, and sperm-zona pellucida binding capacity and (2) to assess their ability to predict fertilization outcome under in vitro fertilization conditions. Semen samples from 44 infertile men were prospectively evaluated for density, morphologic pattern (strict criteria), computerized motion parameters (motility, velocity, and linearity), and hemizona assay (outcome expressed as hemizona assay index), and results were correlated with fertilization outcomes of preovulatory oocytes during in vitro fertilization. Of all sperm parameters, morphologic pattern was the best predictor of the ability of the sperm to bind to the zona pellucida. Hemizona assay index was the best predictor of fertilization rate. Stepwise regression analysis provided a model of hemizona assay index plus motility with highest predictability (R2 = 53.4%). (1) The influence of sperm morphologic pattern as a prognosticator of fertilization outcome is established in major part as a determinant of sperm binding ability to the zona pellucida; (2) the hemizona assay provides a robust index that is highly predictive of the potential of human gametes to achieve fertilization and highlights its use in infertility and contraception testing.


Gonadotropins enhance HLA-DR antigen expression in human granulosa cells. To determine whether HLA-DR expression in the human ovary is controlled by endocrine or immune mediators, we examined the effects of several lectins, lymphokines, and hormones on granulosa cells in vitro. HLA-DR antigens were detected by indirect immunofluorescence on granulosa cells obtained at oocyte retrieval for in vitro fertilization. When cultured with concanavalin A, pokeweed mitogen, phytohemagglutinin, interleukin-1, or interleukin-2, granulosa cells failed to express HLA-DR antigens. Both interferon alfa and interferon beta induced only a modest expression of HLA-DR molecules (4.1% +/- 0.9% and 2.5% +/- 0.8%, respectively) in comparison with the marked increase in HLA-DR expression (25.2% +/- 2.8%), when granulosa cells were exposed to interferon gamma. The gonadotropins follicle-stimulating hormone and human chorionic gonadotropin alone were unable to induce HLA-DR antigens, but acted synergistically with interferon gamma to enhance expression in a dose-dependent manner. The addition of 0.1 and 10 IU/ml of follicle-stimulating hormone with 10,000 IU/ml of interferon gamma significantly increased expression by 11.5% +/- 2.5% and 15.5% +/- 2.7%, respectively, over that of interferon gamma alone. Similarly, 1 and 10 IU/ml of human chorionic gonadotropin produced a 7.8% +/- 2.0% and 12.4% +/- 2.2% increase in HLA-DR expression over baseline. These studies demonstrate that factors from both the immune and endocrine systems can interact to control HLA-DR expression in the ovary.


A human endometrial explant system: validation and potential applications. Our objective was to describe an in vitro explant system to study the regulation of prostaglandin production by human endometrium. Segments of late-luteal-phase endometrium were obtained aseptically at the time of endometrial sampling. The endometrium was cut into 1 mm3 pieces and applied to the polycarbonate membrane of tissue-culture-well inserts for 12-well plates (Costar Transwell cell culture chamber inserts, 0.4 microns pore size). After placing the well inserts, culture medium was carefully applied. The explants were incubated at 37 degrees C in 5% carbon dioxide in air, and the culture medium was changed daily. Electron microscopic examination and lactate dehydrogenase determinations of the explants revealed cellular viability for < or = 5 days of culture. Endometrial explants responded to treatment with interleukin-1 beta and tumor necrosis factor by a concentration-dependent increase in the production of prostaglandin E2. Costimulation of late luteal endometrial explants with interleukin-1 beta (10 ng/ml) and progesterone (10(-6) mol/L) resulted in variable production of prostaglandin E2, suggesting that the histologic examination of the endometrium does not necessarily reflect the functional properties of the endometrium. Our data show that when used with human endometrial tissue this explant system maintains tissue viability and responsiveness for < or = 5 days. This approach to explant methods is simple and provides a flexible model to study the regulation of the production of bioactive substances by human endometrial tissue.


Effects of chronic nocturnal opiate antagonism on the menstrual cycle. Opioids generally suppress gonadotropin secretion. We investigated whether chronic opiate antagonism would affect the occurrence or timing of menstrual cycle events. Opiate blockade was maintained in nine normal volunteers from cycle days 1 through 13 with naltrexone. A double-blind, placebo-controlled crossover design was used. Serum estradiol and progesterone were measured daily. Vaginal ultrasonography was performed daily beginning cycle day 10. Endometrial biopsies were performed 10 to 12 days after ovulation. Nonparametric paired data analysis was used. No significant differences in the following menstrual cycle parameters were detected between naltrexone and placebo cycles: cycle length (25.4 +/- 0.9 vs 25.7 +/- 0.5 days), maximum follicle size (21.5 +/- 1.0 vs 20.8 +/- 1.1 mm), peak serum progesterone (15.8 +/- 1.8 vs 15.2 +/- 2.2 ng/ml), or integrated serum progesterone (108.1 +/- 12.8 vs 101.2 +/- 14.6 ng/ml/day). Histologic endometrial development was normal in all subjects during naltrexone cycles. These data suggest that naltrexone-suppressible opioid activity does not play a significant role in the regulation of follicular events in normal women.


Cocaine impairs gonadotropin secretion in oophorectomized monkeys. Our objective was to determine whether cocaine alters gonadotropin secretion in oophorectomized monkeys. Oophorectomized monkeys with elevated gonadotropin levels were chronically cannulated to allow blood sampling every 15 minutes. Monkeys received either saline solution or 2 or 4 mg/kg cocaine hydrochloride as an intravenous bolus. Other oophorectomized monkeys were pretreated with either saline solution or 4 mg/kg cocaine 2 hours before bolus gonadotropin-releasing hormone administration, and plasma luteinizing hormone and follicle-stimulating hormone levels were measured every 15 minutes for 3 hours. Monkeys were also given either saline solution or 4 mg/kg of cocaine with gonadotropin-releasing hormone simultaneously, and plasma gonadotropin levels were measured every 15 minutes for 3 hours. Serum luteinizing hormone and follicle-stimulating hormone levels were measured by radioimmunoassay. Both doses of cocaine resulted in a significant decrease in luteinizing hormone levels compared with controls. Follicle-stimulating hormone levels were significantly decreased only with the 4 mg/kg dose of cocaine. There was no difference in luteinizing hormone and follicle-stimulating hormone responses to gonadotropin-releasing hormone in the cocaine-treated monkeys compared with saline solution-treated monkeys by using repeated-measures analysis of variance. These findings demonstrate that acute cocaine administration to oophorectomized primates inhibits basal luteinizing hormone-follicle-stimulating hormone secretion but not gonadotropin-releasing hormone-stimulated luteinizing hormone and follicle-stimulating hormone release. In the absence of an effect on gonadotropin-releasing hormone-stimulated gonadotropin release, we conclude that the impaired luteinizing hormone-follicle-stimulating hormone secretion after cocaine administration is due in part to a direct effect of cocaine on gonadotropin-releasing hormone neurons or on hypothalamic neurotransmitter modulation of gonadotropin-releasing hormone release.


Absence of the testicular determining factor gene SRY in XX true hermaphrodites and presence of this locus in most subjects with gonadal dysgenesis caused by Y aneuploidy. The purpose of our study was to discover whether the testicular determining factor gene SRY (sex-determining region on Y) is present or absent in XX true hermaphrodites and in subjects with gonadal dysgenesis caused by Y aneuploidy. We screened five XX true hermaphrodites and 24 subjects with gonadal dysgenesis caused by Y aneuploidy for the presence or absence of SRY. With the polymerase chain reaction technique, the sequence coding the 80 amino acid-conserved motif was amplified. The 0.9 kb Hincll pY53.3 subclone, which covers the open reading frame of SRY, serves as a probe for Southern blot analysis. Test results for all five XX true hermaphrodites were negative for SRY. Conversely, 22 of the 24 individuals with 45,X/46,XY gonadal dysgenesis were positive for SRY, including the 10 subjects with only bilateral streak gonads. The absence of SRY in XX true hemaphrodites and the presence of SRY in 10 subjects with 45,X/46,XY constitution who harbored only bilateral streak gonads seem to indicate that multiple genes are involved in gonadal differentiation.


Increased risk for polycystic ovary syndrome associated with human leukocyte antigen DQA1*0501. The objective of this investigation was to identify genes that confer susceptibility to polycystic ovary syndrome. Nineteen subjects with polycystic ovary syndrome, hirsutism, and elevated plasma androgen levels and 46 fertile, female control subjects were studied. Alleles at the human leukocyte antigen DQA1 locus were identified with dot-blot hybridizations with allele-specific oligonucleotide probes. Associations between human leukocyte antigen DQA1 alleles and polycystic ovary syndrome were examined with logistic regression analysis. The frequency of the human leukocyte antigen DQA1*0501 was 0.50 and 0.26 in subjects with polycystic ovary syndrome and control subjects, respectively (corrected for multiple comparisons, p = 0.067). Homozygosity for this allele was also increased among subjects with polycystic ovary syndrome (p = 0.054). The odds ratios for having polycystic ovary syndrome associated with the *0501 allele and the *0501/*0501 homozygous genotype were 2.8 and 5.8, respectively. These data suggest that a polycystic ovary syndrome susceptibility allele is linked to human leukocyte antigen and that the susceptibility allele is recessive.


Does ethnicity influence the prevalence of adrenal hyperandrogenism and insulin resistance in polycystic ovary syndrome? Our purpose was to determine the prevalence of adrenal hyperandrogenism and insulin resistance in patients with hyperandrogenic chronic anovulation, also called polycystic ovary syndrome, living in the United States, Italy, and Japan. Seventy-five women with polycystic ovary syndrome, 25 each from the United States, Italy, and Japan, and 10 ovulatory controls were studied. Hirsutism, obesity, and the presence of cystic ovaries were assessed, as were blood levels for estrogen, luteinizing hormone, testosterone, adrenal androgens, and insulin. All patients received an insulin tolerance test to assess insulin resistance. Women from Japan were less obese (p < 0.05) and did not have hirsutism, although the percentage of cystic ovaries (68% to 80%) was comparable. Serum luteinizing hormone, testosterone, and estradiol were similar, but levels of 3 alpha-androstanediol glucuronide, which was elevated in women from the United States and Italy, was normal in women from Japan. The adrenal androgens, dehydroepiandrosterone sulfate and 11 beta-hydroxyandrostenedione were elevated in 48% to 64% of the patients and by a similar percentage in the three groups. Fasting insulin was elevated in all groups, but was significantly higher in women from the United States and Italy compared with women from Japan (p < 0.05). However, insulin resistance as assessed by dissociation constant of insulin tolerance test values was significantly elevated but similar in the three groups and occurred in 68% to 76% of patients. In polycystic ovary syndrome, although obesity and hirsutism vary according to dietary, genetic, and environmental factors, the prevalence of adrenal androgen excess and insulin resistance appear to be fairly uniform. These results suggest that these factors may be involved in the pathophysiologic features of the disorder.


Lipoprotein abnormalities in hirsute women. II. Compensatory responses of insulin resistance and dehydroepiandrosterone sulfate with obesity. The purpose of our study was to further understand interrelationships between insulin, androgens, obesity, and apolipoprotein metabolism. In this University cross-sectional observational study for hypothesis generation, 53 women with hirsutism-hyperandrogenism were ternately divided according to body mass index into the following groups: 22 to 28 (n = 17), 29 to 36 (n = 19), 37 to 61 (n = 17), and 16 controls. Mean hormone and clinical parameters were compared by using the Student t test, analysis of variance, and Pearson correlation. Dehydroepiandrosterone sulfate correlated negatively with fasting insulin (r = 0.4, p < 0.05) and reached significance in those most obese. Unbound testosterone concentrations were 24.7, 38.9, and 31.9 ng/dl, respectively. Dehydroepiandrosterone sulfate concentrations were 2.8, 2.3, and 2.3 micrograms/dl, respectively; their ratios were 13.4, 18.6, and 20.4, respectively, even though mean fasting insulin concentrations (reflecting insulin resistance) were 13.0, 20.4, and 42.6 microU/ml, respectively. Although more atherogenic apolipoprotein profiles and higher fasting insulin concentrations were found with greater body weight, lipid-insulin interrelationships may reflect dehydroepiandrosterone sulfate interaction. We hypothesize that dehydroepiandrosterone sulfate dampens the effect of insulin resistance, which in turn is reflected in apolipoprotein profiles in women with hirsutism/hyperandrogenism.


Increased circulating levels of immunoreactive beta-endorphin in polycystic ovary syndrome is not caused by increased pituitary secretion. Our purpose was to investigate the source and role of elevated levels of immunoreactive beta-endorphin in polycystic ovary syndrome. We wished to determine whether immunoreactive beta-endorphin secretion in patients with polycystic ovary syndrome is influenced by body weight and whether the pituitary release of immunoreactive beta-endorphin with corticotropin-releasing hormone is related to luteinizing hormone levels or adrenal androgen secretion. Eighteen patients with polycystic ovary syndrome and 10 ovulatory controls were studied. Each subject received 1 microgram/kg intravenous corticotropin-releasing hormone and an oral glucose tolerance test on alternate days. Levels of plasma immunoreactive beta-endorphin, corticotropin, luteinizing hormone, cortisol, adrenal androgens, and insulin were measured. Although immunoreactive beta-endorphin levels were elevated in patients with polycystic ovary syndrome (p < 0.01), incremental responses after corticotropin-releasing hormone were similar to controls and were not influenced by body weight. Serum luteinizing hormone levels were not affected by corticotropin-releasing hormone and did not correlate with immunoreactive beta-endorphin levels. Adrenal androgen responses after corticotropin-releasing hormone were increased in patients with polycystic ovary syndrome (p < 0.01) but were not correlated with immunoreactive beta-endorphin secretion. After oral glucose was given, elevated fasting insulin levels increased significantly in patients with polycystic ovary syndrome (p < 0.01), as did immunoreactive beta-endorphin levels (p < 0.05). The increases in insulin and immunoreactive beta-endorphin levels were correlated (p < 0.05). Pituitary secretion of immunoreactive beta-endorphin is normal in patients with polycystic ovary syndrome, and pancreatic secretion appears to be increased. Corticotropin-releasing hormone does not influence luteinizing hormone levels, and adrenal androgen sensitivity is not influenced by immunoreactive beta-endorphin secretion.


Increased bioactive luteinizing hormone levels in postmenopausal women with endometrial cancer. The purpose of our study was to investigate the secretion of immunoactive and bioactive luteinizing hormone in postmenopausal women with endometrial cancer. Seventeen postmenopausal women with endometrial cancer and nine without cancer were studied. Blood samples were collected at 15-minute intervals for 360 minutes. All samples were assayed for bioactive luteinizing hormone by rat interstitial cell testosterone assay and for immunoactive luteinizing hormone by radioimmunoassay. Serum pooled from 6-hour sampling was assayed for follicle-stimulating hormone, total estradiol, unbound estradiol, and estrone. Patients with endometrial cancer had significantly (p < 0.01) higher bioactive luteinizing hormone levels (mean +/- SE 276 +/- 26 IU/L) as compared with those of control women (144 +/- 18 IU/L). Bioactive/immunoactive ratios of luteinizing hormone were significantly higher (p < 0.01) in women with cancer (5.8 +/- 0.7) than in those without cancer (2.5 +/- 0.5). There was a significant (p < 0.001) positive correlation (r = 0.582) between unbound estradiol levels and bioactive luteinizing hormone concentrations. There is an increase in bioactive luteinizing hormone secretion in postmenopausal women with endometrial cancer. This could lead to an increase in ovarian androgen production resulting in increased prehormone availability for estrogen formation from peripheral conversion.


Behavioral stress responses in premenopausal and postmenopausal women and the effects of estrogen. Our purpose was to determine the pattern of reactivity to stress in premenopausal and postmenopausal women and to assess the effects of estrogen. A behavioral stress test was given to premenopausal (n = 13) and postmenopausal women (n = 36). Biophysical and neuroendocrine responses were measured during and on completion of the stress test. The postmenopausal women were then randomized to placebo or transdermal estradiol treatment for 6 weeks, at which time another behavioral stress test was given. Stress reactivity to math and speech tasks elicited significantly greater systolic blood pressure responses in postmenopausal women compared with premenopausal women (p < 0.05). On retesting, significant biophysical responses that were present during the initial stress testing were still present (p < 0.05) in the placebo group but were blunted with estrogen treatment. Plasma corticotropin, cortisol, androstenedione, and norepinephrine increased during testing to a similar degree in premenopausal and postmenopausal women; this response was maintained after placebo treatment. Postmenopausal women treated with estrogen had blunted responses. Significant differences in responses to psychologic stress exist in premenopausal and postmenopausal women. The lack of adaptation may account in part for the increased risk of cardiovascular disease in postmenopausal women. Estrogen appears to blunt the stress-induced response.


Aldose reductase inhibition prevents galactose-induced ovarian dysfunction in the Sprague-Dawley rat. Our objective was to determine whether impaired ovarian function induced by short-term creation of a galactosemic state in the rat might be prevented by the coadministration of an aldose reductase inhibitor. Prepubertal Sprague-Dawley rats were fed four different diets including (1) control, (2) 40% galactose, (3) 40% galactose and an aldose reductase inhibitor, and (4) an aldose reductase inhibitor with the control diet. Percentage germinal vesicle breakdown, postovulatory oocyte quantities, hormonal parameters, ovarian histologic evaluation, and ovarian galactitol concentrations were determined. The galactose-fed animals (group 2) had decreased germinal vesicle breakdown (47%) versus control (69%, p < 0.05). Galactose-exposed animals had significantly decreased quantities of postovulatory eggs (6.4 per animal) after menotropin ovarian stimulation in comparison with controls (14.1, p < 0.01). In rats exposed to high dietary levels of galactose (group 2) ovarian galactitol concentrations were significantly higher (protein 42.12 mumol/gm versus 0.0 for controls, p < 0.005). When galactose-fed animals received the aldose reductase inhibitor, ovarian accumulation of galactitol was significantly reduced and the observed detrimental effects on the oocyte were prevented. Galactitol accumulation or metabolic flux through aldose reductase in galactosemic rodents may be involved in the demonstrated ovarian dysfunction.


Immunologic tests in the evaluation of reproductive disorders: a critical review. The purpose of this study was to evaluate the clinical usefulness of immunologic assays proposed to assist in the diagnosis of alloimmune causes of recurrent spontaneous abortion. Human leukocyte antigen typing, maternal antipaternal lymphocytotoxic antibody testing, and mixed lymphocyte culture assays were performed on 609 couples with recurrent spontaneous abortion, 92 infertile couples, and 43 fertile couples. The frequency of HLA antigen sharing and the presence of lymphocytotoxic antibodies and mixed lymphocyte culture inhibitors was compared among the populations. Sharing of two or more HLA-A, HLA-B, HLA-C, or HLA-DR antigens was observed in 41% (252/609) couples with recurrent spontaneous abortion and in 34% (31/92) with infertility compared with 63% (27/43) of fertile couples. The frequency of lymphocytotoxic antibodies to paternal cells was significantly greater in the fertile population (27/143, 63%) (p < 0.0001) than in couples with recurrent spontaneous abortion (160/609, 26%) and in infertile couples (7/92, 8%). Mixed lymphocyte culture inhibitors were also more prevalent in sera from fertile women (19/43, 44%) compared with those with recurrent spontaneous abortion (122/609, 20%) and infertile women (4/92, 4%). Human leukocyte antigen sharing does not predict pregnancy outcome, and the presence of lymphocytotoxic antibodies and mixed lymphocyte culture inhibitors is a function of the number and duration of pregnancies. More sensitive and specific assays are needed to identify alloimmunologic causes of reproductive disorders.


Photodynamic therapy of human ovarian epithelial carcinoma, OVCAR-3, heterotransplanted in the nude mouse. Our objective was to develop an animal model for the study of photodynamic therapy in the treatment of human ovarian epithelial carcinoma. The human ovarian carcinoma OVCAR-3 was heterotransplanted into nude, athymic mice and treated with photodynamic therapy consisting of the hematoporphyrin derivative Photofrin II 10 ng/kg and argon-pumped dye laser light at 630 nm (200 J/cm2). Growth of tumors on one side of seven mice (treated tumors, n = 7) was compared with contralateral tumors (control tumors, n = 8) not exposed to laser. Hematoporphyrin derivative uptake was determined in tumor and other tissues. Photodynamically treated tumors were completely ablated, and all remained absent. Hematoporphyrin derivative uptake was nonselective for tumor compared with other tissues. This model provides reproducible parameters for the study of photodynamic therapy in the treatment of gynecologic malignancies and demonstrates the need for methods to increase selective uptake of hematoporphyrin derivatives.


Endometrial ablation by means of photodynamic therapy with photofrin II. Photodynamic therapy is a technique in which tissue is irradiated with light after the use of a photosensitizing drug that produces singlet oxygen, which has a cytotoxic effect. The feasibility of using photodynamic therapy with photofrin II for endometrial ablation was studied. Fifty-eight rabbits were studied. Preferential uptake of photofrin II by endometrial tissue, compared with the myometrium, was established by drug extraction and fluorescence microscopy after administration of photofrin II intravenously. Dosimetry for endometrial ablation was established by administering photofrin II in 1, 2, 5, and 10 mg/kg doses and laser light (630 nm) at radiant exposures of 100 and 200 J/cm2. Histologic examination was performed at 24 hours, 5 days, and 10 days after treatment. There were two control groups. One group received laser light but no photofrin II, and the other received photofrin II without laser light. The concentration of photofrin II was three times higher in the endometrium than in the myometrium at doses of 1 and 2 mg/kg. Fluorescence microscopy of frozen sections of endometrium and myometrium showed a predominantly perivascular fluorescence from photofrin II. A dose of 1 and 2 mg/kg and a flow of 100 J/cm2 was adequate for endometrial ablation in rabbits. At 24 hours after treatment there was extensive hemorrhage and evidence of cell death in the entire endometrium and mild hemorrhage in 10% to 50% of the inner circular layer of the myometrium. At 5 days after treatment necrosis of the entire endometrium and the inner half of the myometrium was seen, but the outer half of the myometrium and the serosa were normal. There were no cases of uterine perforation. Similar results were seen at 10 days after treatment, except for the additional presence of inflammatory cells. Neither control group (drug without light, light without drug) showed any injury to the endometrium at 24 hours. We conclude that endometrial ablation can be effectively achieved in rabbits by means of photodynamic therapy with photofrin II without significant complications.


Quantitation of tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6 in the effusions of ovarian epithelial neoplasms. Our objective was to determine the presence and quantities of multifunctional cytokines in cyst and ascites fluids obtained from patients with ovarian cancer. Cyst and ascites fluids obtained from 35 patients with ovarian epithelial neoplasms were analyzed for the multifunctional cytokines tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6. The fluids were classified on the basis of the pathologic diagnosis of the tumor tissue. Data were evaluated by analysis of variance. The fluids from patients with papillary forms of adenocarcinoma had high levels of all three cytokines when compared with all other pathologic groups. Interleukin-6 was significantly higher than tumor necrosis factor-alpha or interleukin-1 beta in fluids from all diagnostic categories (p < 0.01). Interleukin-6 levels were significantly higher in fluids from patients with papillary serous cystadenocarcinoma (817 +/- 137 pg/ml) and papillary adenocarcinoma (838 +/- 171 pg/ml) (p < 0.01). Interleukin-1 beta was significantly elevated (p < 0.01) in neoplastic effusions from papillary serous cystadenocarcinomas (53 +/- 8 pg/ml), mucinous cystadenomas (51 +/- 12 pg/ml), and endometrioid carcinomas (54 +/- 18 pg/ml). Tumor necrosis factor-alpha was highest in fluids from patients with papillary adenocarcinoma (46.2 +/- 22.8 pg/ml); however, these levels were not significantly different from the mean quantities of tumor necrosis factor-alpha in other fluids. The detection of interleukin-1 beta and interleukin-6 in neoplastic effusions provides possible evidence for a host immune response to ovarian cancer. These multifunctional cytokines have been implicated in growth stimulation and cytotoxicity of ovarian tumor cells.


Telomere reduction in endometrial adenocarcinoma. Some of the genomic instability that is observed in solid tumors may be due to the loss of telomeric sequences. These experiments were designed to compare the number of telomeric repeat sequences in endometrial adenocarcinoma with that found in adjacent normal tissue. Deoxyribonucleic acid was extracted from normal and malignant uterine tissue of 11 patients undergoing hysterectomy for treatment of endometrial adenocarcinoma and also from five endometrial carcinoma cell lines. The relative number of telomeric repeat sequences in each sample was measured by hybridization of these deoxyribonucleic acids to a probe specific for the human telomeric repeat. Hybridization signals were quantified by autoradiography and a beta-particle detection system. A reduction of telomeric repeat sequences in tumor versus normal tissue was found in 10 of 11 cases. Telomere reduction was also seen in endometrial carcinoma cell lines. Telomere reduction is a genetic characteristic of many endometrial tumors. Telomere reduction may contribute to the genesis and progression of endometrial carcinoma, or it may be a secondary effect of the tumorigenesis process.


Culture characteristics of human endometrial glandular epithelium throughout the menstrual cycle: modulation of deoxyribonucleic acid synthesis by 17 beta-estradiol and medroxyprogesterone acetate. The purpose of this study was to evaluate the culture characteristics and cell proliferation of human endometrial glandular epithelial cells in primary culture on extracellular matrix and to evaluate sex steroid modulation of this process. We examined the culture characteristics in 53 endometrial gland preparations obtained throughout the menstrual cycle and determined the incorporation of tritiated thymidine in endometrial epithelial cells as a measure of deoxyribonucleic acid synthesis (cell proliferation) in the presence and absence of 17 beta-estradiol and medroxyprogesterone acetate. Good culture maintenance of endometrial epithelial monolayers on extracellular matrix was observed from glands derived from proliferate phase endometrium (21 of 23), whereas poor adherence and culture maintenance was observed in all 30 of specimens from the secretory phase. With 17 beta-estradiol treatment for 22 hours, tritiated thymidine incorporation was not different from control, but medroxyprogesterone acetate treatment for 22 hours was associated with diminished tritiated thymidine incorporation by 36% (p < 0.004). When 17 beta-estradiol (10(-8) mol/L) was included in the incubation medium from the time of initial culture, tritiated thymidine incorporation on day 4 was 40% of control (p < 0.006), and tritiated thymidine was not suppressed further by 22 hours of treatment with medroxyprogesterone acetate (10(-7) mol/L). In spite of cellular spread of endometrial epithelial cells to subconfluence, deoxyribonucleic acid content per well did not increase over time in culture. We conclude that exposure of endometrial glands to progesterone in vivo inhibits adherence and the establishment of cell monolayers cultured on a thin layer of extracellular matrix. Because 17 beta-estradiol treatment in culture for 22 hours does not increase tritiated thymidine incorporation, it is possible that 17 beta-estradiol exerts its proliferative effect on the endometrial epithelium by an indirect action mediated by stromal cells. The effects of 17 beta-estradiol and medroxyprogesterone acetate on deoxyribonucleic acid synthesis in endometrial epithelial cells may represent an action on a stem cell population of dividing cells or a terminally differentiated cell population that undergoes programmed cell death.


Beta-carotene levels in exfoliated cervicovaginal epithelial cells in cervical intraepithelial neoplasia and cervical cancer. The purpose of this study was to measure beta-carotene levels in exfoliated epithelial cervicovaginal cells collected by a lavage technique in normal women and patients with histopathologically diagnosed cervical intraepithelial neoplasia and cervical cancer. In a cross-sectional sampling of women (n = 105), cervicovaginal cells and plasma beta-carotene levels were assayed with high-pressure liquid chromatography. In addition, beta-carotene levels were measured in exfoliated epithelial samples of cervicovaginal cells obtained from women (n = 24) enrolled in an ongoing oral beta-carotene supplementation clinical trial. Cervicovaginal cells and plasma beta-carotene levels were found to be significantly decreased in women with cervical intraepithelial neoplasia and cervical cancer as compared with controls (p < 0.0001, analysis of variance). Retinol levels in cervicovaginal cells were undetectable. The beta-carotene levels in cervicovaginal cells were markedly increased in the majority of patients (79%) after oral supplementation as compared with baseline levels in women enrolled in the beta-carotene clinical trial. The study demonstrates that changes of in situ cellular beta-carotene concentrations are measurable in samples of exfoliated epithelial cells obtained by a noninvasive saline lavage harvesting technique. The current findings further support our previous hypothesis that beta-carotene deficiency may have an etiologic role in the pathogenesis of cervical intraepithelial neoplasia and/or cervical cancer.


Factors influencing the exfoliation of cervicovaginal epithelial cells. Our objective was to investigate the association of smoking and exfoliation of cervicovaginal epithelial cells while controlling for other factors that may potentially influence cell exfoliation (e.g., presence of cervical intraepithelial neoplasia or koilocytes, the use of oral contraceptives, age, and the phase of the menstrual cycle). Cervicovaginal lavage specimens and epidemiologic questionnaires were obtained with informed consent from 190 women. The cervicovaginal lavage samples were processed to separate other contaminants. The number of squamous epithelial cells counted was expressed as cells per milliliter of lavage. Multiple linear regression analysis revealed that the number of exfoliated epithelial cells was significantly higher in smokers (p < 0.01) and also in women with cervical intraepithelial neoplasia (p < 0.05). The other studied variables had no detectable effect. The findings suggest that smoking or the presence of cervical intraepithelial neoplasia may induce an acceleration in the exfoliation of cervicovaginal epithelial cells. This may alter cell maturation and may be a factor in the oncogenic process.


Cytoskeletal immunohistochemistry of central neurocytomas. Central neurocytomas are rare intraventricular tumors. Patients with such tumors have a favorable prognosis after surgical removal. These tumors may be misdiagnosed as neuroblastomas or gliomas, risking the complications of adjuvant therapy. Diagnosis of central neurocytoma requires that the tumor shows the ultrastructural features of mature neuronal differentiation, including the presence of synapses and dense-core and clear vesicles in addition to profiles of neuritic processes with microtubules. The cytoskeletal phenotype of central neurocytomas has not been previously characterized, but it may facilitate their definitive recognition when ultrastructural examination is not possible. Ten central neurocytomas were examined by immunohistochemistry for phosphorylation-dependent/independent neurofilament epitopes, neuron-associated class III beta-tubulin, microtubule-associated proteins (MAP2, tau), and glial fibrillary acidic protein (GFAP). The neuronal nature of all neoplasms was documented by immunoreactivity for synaptophysin in nine tumors and for phosphorylation-independent neurofilament-H/M in the remaining case. Electron microscopy in four cases showed synapses and dense core vesicles. All tumors were immunoreactive for class III beta-tubulin and MAP2, which were seen in cytoskeletal structures by immunoelectron microscopy. Two thirds of the cases were immunohistochemically positive for neurofilament epitopes. None of the tumor cells displayed GFAP immunoreactivity, although reactive astrocytes were present. These data suggest that central neurocytomas may be recognized by synaptophysin immunoreactivity and that the expression of cytoskeletal epitopes indicates that these tumors are well-differentiated neuronal neoplasms.


Acinic cell carcinoma of the lung ("Fechner tumor"). A clinicopathologic, immunohistochemical, and ultrastructural study of five cases. The clinical, pathological, immunohistochemical, and ultrastructural features in five cases of primary acinic cell carcinoma of the lung are presented. The patients' ages ranged from 44 to 75 years (mean, 56); four were women and one a man. The lesions were discovered incidentally on routine chest x-ray and ranged from 1.2 to 4 cm in greatest diameter. Three tumors were located in the right middle lobe, one in the right upper lobe, and one in the left upper lobe. In three cases, the lesions presented as asymptomatic subpleural nodules in proximity to secondary bronchi, one case presented as an endobronchial tumor that led to obstructive symptoms, and one case as a well-circumscribed deep parenchymal nodule. Histologically, the tumors were composed of clear cells with abundant granular cytoplasm growing as solid sheets with focal acinar, microcystic, and papillocystic areas. Immunohistochemical stains showed strong positivity of the tumor cells for low-molecular-weight cytokeratins and epithelial membrane antigen (EMA). Focal weak cytoplasmic positivity was observed in three cases with alpha-1-antichymotrypsin and in one case with amylase. Stains for vimentin, S-100 protein, chromogranin, and lysozyme were negative in all cases examined. Electron microscopy performed in four cases showed abundant zymogen-type cytoplasmic granules of variable electron density characteristic of acinar-type secretory cells. All patients were treated by lobectomy alone. Follow-up of 3 to 10 years in four cases revealed that all patients were alive and well, with no evidence of recurrence or metastases. Because of their relatively indolent behavior and favorable prognosis, primary acinic cell carcinoma of the lung must be distinguished from other primary and metastatic clear cell tumors of the lung.


Well-differentiated liposarcoma (atypical lipoma) of deep soft tissue of the extremities, retroperitoneum, and miscellaneous sites. A follow-up study of 92 cases with analysis of the incidence of "dedifferentiation". Ninety-two cases of well-differentiated liposarcoma of deep soft tissue of the extremity, retroperitoneum, and groin with follow-up information of at least 2 years and no evidence of dedifferentiation at the time of diagnosis were studied to determine their long-term behavior. The tumors occurred most commonly in the muscles of the extremity (46 cases), followed by the retroperitoneum (23 cases), groin (14 cases), and miscellaneous sites (nine cases). Tumors in the retroperitoneum recurred in nearly all cases (21 of 23 cases), occasionally caused patient death, and dedifferentiated in four cases (median time to dedifferentiation, 8 years). Tumors in the groin had a similar high recurrence rate (11 of 14 cases), caused death of patients (two of 14 cases), and dedifferentiated (four of 14 cases). In contrast, those in the extremity recurred less frequently (20 of 46 cases) and had no disease-related mortality. Three of 46, however, underwent dedifferentiation (median time to dedifferentiation, 7 years). Of the 11 cases that underwent dedifferentiation, the interval between diagnosis and dedifferentiation ranged from 2 to 18 years (median time, 9 years; average time, 11 years). Six of the 11 patients showed dedifferentiated foci in the first recurrence, and three died of metastatic disease. Our study indicates that the behavior of well-differentiated liposarcomas is strongly influenced by location. Although the prevailing view is that dedifferentiation is restricted to tumors of the retroperitoneum, our study indicates that it is not a site-specific phenomenon, but is more likely a time-dependent phenomenon seen in situations with a high likelihood for clinical persistence of disease for a long period. Dedifferentiation identifies a tumor that is potentially more aggressive; yet the progression of the disease following dedifferentiation may be highly variable and probably depends on a number of factors, including the amount of dedifferentiation and type of therapy. Although atypical lipoma is an acceptable term for well-differentiated liposarcomas of the subcutis, it fails to convey the potentially life-threatening properties of retroperitoneal tumors. For these lesions as well as those in the deep soft tissues of the extremity, retention of the term well-differentiated liposarcoma is advocated.


Aggressive angiomyxoma. A report of four cases occurring in men. Aggressive angiomyxoma is a distinctive, locally aggressive but nonmetastasizing soft tissue tumor of the pelvic soft tissues and perineum. This rare tumor occurs almost exclusively in adult women. We report four cases occurring in adult men, one each located in the scrotum, inguinal region, spermatic cord, and pelvis. The tumors were infiltrative, and were composed of fibromyxoid matrix sparsely populated by bland-looking spindled and stellate cells with delicate cytoplasmic processes. There were haphazardly scattered small and large blood vessels, some of which exhibited hypertrophy or hyalinization of the wall. Immunohistochemically, the stromal cells stained consistently for vimentin and variably for muscle-specific actin, but not alpha-smooth muscle actin, desmin, and S-100 protein. None of the four tumors recurred in follow-up times from 11 months to 6 years, although two previously reported cases in men recurred. This uncommon tumor occurring around the genital region in men merits wider recognition because of its potential for recurrence. It should be distinguished from benign tumors with low risk of recurrence on one hand, and from malignant myxoid tumors with metastatic potential on the other.


Plasmacytic differentiation in parafollicular (monocytoid) B-cell lymphoma. A study of 12 cases. Parafollicular (or monocytoid) B-cell lymphoma (PBCL) is a recently described low grade lymphoma. The relationship of parafollicular B cells to other B lymphocytes is not known, but the authors observed plasmacytic differentiation in the initial case of PBCL. In this report 12 cases of PBCL were studied by light microscopy and immunophenotypic analysis, and plasmacytic differentiation was found in four cases. This plasmacytic differentiation and the anatomic relationship of the neoplastic cells to reactive follicular centers suggest a functional relationship between these cell types.


Prethymic adult lymphoblastic lymphoma. A clinicopathologic and immunohistochemical analysis. The clinical, histologic, and immunohistologic features of three cases of pre-T-cell (CD7+/CD2-) lymphoblastic lymphoma in adults are reported. The patients were adults over age 50 years who had a relatively indolent nodal disease, partial involvement of lymph nodes, and primitive immunophenotype. The phenotype of the three cases was TdT+, HLA-DR+, CD34+, CD71+, CD38+, and CD7+, most resembling the normal prothymocyte, and in contrast to normal thymocytes, which generally coexpress CD1+, CD4+, and CD8+. The prethymic T-cell character was further supported by germline T-cell receptor beta and gamma chain genes. In contrast to most reported cases with this early immunophenotype, these patients had nodal disease but not peripheral blood involvement. Two of the three cases were associated with Langerhans' cell histiocytosis (histiocytosis X), a previously unreported association. Because of the Langerhans' cell histiocytosis and the relatively indolent clinical presentation, the differential diagnosis in all cases included both a benign process and a lower-grade lymphoma. Recognition of this unusual form of adult lymphoblastic lymphoma is essential for correct diagnosis and treatment.


Uterine carcinomas simulating microglandular hyperplasia. A report of six cases. Six uterine adenocarcinomas, one cervical and five endometrial, that simulated microglandular hyperplasia histologically are described. The cervical neoplasm occurred in a 34-year-old woman who had been taking oral contraceptives. The five endometrial cancers occurred in postmenopausal patients 57 to 69 years of age; two of them were receiving, or had been receiving, premarin and provera and two were receiving only premarin. The patient with the cervical neoplasm had an abnormality observed on pelvic examination. The five postmenopausal patients presented because of vaginal spotting, bleeding, or discharge; their carcinomas were discovered in endometrial curettage specimens. On initial pathologic examination two specimens were misinterpreted as benign and in the remaining four cases there was uncertainty as to whether the lesion was benign or malignant. All the neoplasms had conspicuous microglandular patterns, with the gland lumens typically containing eosinophilic mucinous secretion and numerous acute inflammatory cells, which were also characteristically prominent in the stroma. Subsequent hysterectomies in all the cases showed residual adenocarcinoma with more typical features. Three of the endometrial tumors were mucinous adenocarcinomas and two mixed mucinous and endometrioid adenocarcinomas; the cervical tumor was a moderately differentiated adenocarcinoma of endocervical type. These cases illustrate that microglandular hyperplasia should be diagnosed with caution in a postmenopausal patient, particularly if lesional tissue is present in an endometrial curettage specimen, and, rarely, cervical adenocarcinomas have foci that simulate microglandular hyperplasia.


Pancreas rejection. Significance of histopathologic findings with implications for classification of rejection. To determine the significance of various histopathologic features of pancreatic rejection, we reviewed the pathology of 53 biopsies taken to rule out rejection [32 bladder drained, (BD); 18 non-BD]. Twenty-six biopsies from 23 patients with allografts which ultimately failed (FLD) (7 BD, 16 non-BD) were compared with 27 biopsies from 27 patients with allografts which continue to function (FXN) (25 BD, 2 non-BD). The groups are similar in regard to age, sex, and time after transplant to biopsy. The mean follow-up is 13 months for FLD grafts versus 35 months for FXN grafts (p < 0.0001). In BD grafts, decreases in urine amylase usually led to biopsy, while in non-BD grafts, hyperglycemia usually prompted biopsy. More patients with ultimately FLD organs (17 of 26) presented with elevated blood glucose (BG) than patients with FXN grafts (2 of 27) (p < 0.0001). Multiple histologic features were examined related to the acinar tissue, pancreatic ducts, islets, vessels, and nerves. Features which strongly correlated with a negative outcome included moderate to severe inflammation of acinar tissue (p < 0.0001), acinar tissue loss and fibrosis (p < 0.0087) and vascular luminal narrowing due to chronic rejection (p < 0.003). Twenty-one pancreases showed chronic rejection and were treated with OKT3 or anti-lymphocytic globulin (ALG), six of these continue to function 1.6-9 years after biopsy, including two who presented with elevated BG levels. A normal biopsy was found in nine pancreases, all of which continue to function. Vasculitis was only seen in biopsies with moderate to severe inflammation, whereas endothelialitis was also seen in association with mild inflammation, suggesting that vasculitis is a more aggressive lesion. A rejection classification is proposed with endothelialitis partly defining mild rejection and vasculitis defining severe rejection. We conclude that several biopsy features and elevation of BG are strongly correlated with a high probability of failure; however, antirejection therapy is justified because recovery of function occurs in some cases. A normal biopsy obviates the need for therapy and predicts a good outcome, as do mild histological findings of rejection.


Basic epidemiology of tuberculosis in Peru: a prevalence study of tuberculin sensitivity in a Pueblo joven. Tuberculosis continues to cause significant morbidity and mortality in developing nations. As a first step in defining the magnitude of the problem in Peru, we determined the prevalence of tuberculin sensitivity in an age-stratified, community-based population on the outskirts of Lima in December 1990. The overall prevalence of 10 mm or more induration in 368 individuals was 34%. When stratified by age, the prevalence was 12% in the 0-1-year-old group, 18% in the 2-4-year-old group, 24% in the 5-14-year-old group, 60% in the 15-24-year-old group, and 68% in the > or = 25-year-old group. Vaccination with bacillus Calmette-Guerin (87% of the study population) caused significant increases in weak (5-9 mm) reactions to purified protein derivative, but did not cause strong (10 mm or more) reactions. The prevalence of tuberculous infection in this population is higher than that previously reported in Peru and in most other high-risk populations. Unfortunately, the current political and economic situation in Peru makes it difficult to implement public health measures to prevent infection and progression of infection to disease.