Simultaneous measurement of adenosine and hypoxanthine in human umbilical cord plasma using reversed-phase high-performance liquid chromatography with photodiode-array detection and on-line validation of peak purity.

A new, robust and sensitive reversed-phase high-performance liquid chromatographic method was developed for concomitant measurement of plasma concentrations of the ATP catabolites adenosine and hypoxanthine in human umbilical cord blood. Deproteinized cord plasma was chromatographed on Hypersil C18 columns, using UV photodiode-array detection, spectral analysis of peaks and on-line confirmation of peak purity. Elution with a gradient of acetonitrile-tetrahydrofuran in ammonium dihydrogen phosphate buffer pH 4.7, yielded sharp, well-resolved peaks of adenosine and hypoxanthine within 16 min. Peak areas were quantified from external calibration curves and converted to plasma concentrations via cord blood hematocrits. In seven deliveries, gestational ages 32-40 weeks, adenosine (range, 0.1-2.1 microM) was less than hypoxanthine (range, 1.6-18.5 microM) in the same cord plasma sample. Arteriovenous levels of each purine were similar, except in an abruptio placenta delivery.

Resolution of unilateral fetal hydrothorax with associated non-immune hydrops after intrauterine thoracentesis.

Congenital hydrothorax has been successfully managed in utero by two different treatment modalities: thoracentesis and pleuroamniotic shunting. Unfortunately, there is a paucity of data as to which method is better for the management of this problem. This case report supports the use of thoracentesis as the initial procedure for primary fetal hydrothorax. We report a pregnancy complicated by primary fetal hydrothorax with non-immune hydrops that completely resolved after two thoracenteses in the early third trimester. Complete resolution was maintained throughout pregnancy without the need for further antenatal or neonatal intervention, i.e. further thoracentesis, pleuroamniotic shunt placement, intubation, or chest and/or abdominal tube placement. The child was delivered at 42 weeks and is doing well without problems at 1 year of age. We believe that intrauterine thoracentesis should be the initial procedure of choice for the treatment of primary fetal hydrothorax with mediastinal shift, and pleuroamniotic shunting should be reserved for cases that require repetitive thoracenteses.

Abruptio placentae and disseminated intravascular coagulation: use of thrombelastography and sonoclot analysis.

We report a case in which thrombelastography (TEG) and Sonoclot analysis were used for diagnosis and treatment decisions in a patient with abruptio placentae and disseminated intravascular coagulation. In addition to providing enough information for evaluation and treatment of this patient, the TEG and Sonoclot results were obtained more quickly, with smaller blood samples and with less expense to the patient in comparison to the standard coagulation tests in our institution. Although the TEG and Sonoclot results were assessed independently of the other test and each provided similar information in this case, the TEG may be the preferred tool in evaluating coagulopathies, based on computerization and strong literature support.

Hyperglycemia delays rostral initiation sites during neural tube closure.

Neural tube defects contribute greatly to perinatal loss, physical handicap, mental retardation and other developmental defects, yet the mechanisms through which they occur are poorly understood. One hindrance to the study of these defects at the cellular and molecular levels is the low frequency with which they arise in susceptible animals. The present study utilizes a culture system for the study of rodent exencephaly, an animal model of human anencephaly, in which a high frequency of affected animals are obtained by culture in hyperglycemic rat serum. Rat embryos were dissected at day 9.5 from timed-pregnant Sprague-Dawley dams and cultured under standard conditions developed by New [Biol. Rev. (1978) 53,81-122]. Embryos cultured under elevated glucose conditions are able to close the caudal neural tube with the failure of neural tube closure limited to the rostral neuralepithelium. In this report we present the novel finding that, although at the end of culture frequently only the hindbrain region remains open, the normal sequence of events expected during rostral closure anterior to the hindbrain is markedly delayed. In embryos cultured in hyperglycemic serum, both rostral initiation sites II and III are significantly delayed. The degree of delay increases with increasing glucose concentration in the culture medium. These studies support the use of this defined in vitro model of anencephaly for studies of the molecular and cellular bases underlying the failure of hindbrain closure and demonstrate that sufficient numbers of affected animals can be produced to obtain significant results.(ABSTRACT TRUNCATED AT 250 WORDS)

Terminal deletion of 7q presenting in utero with a truncus arteriosus and nonimmune hydrops.

Terminal deletion of 7q presents with variable anatomical and developmental findings. This case is the first reported in utero diagnosis based on cytogenetic findings and in utero demonstration of resolving congestive heart failure due to a truncus arteriosus.

Development of the Eagle-Barrett (prune belly) syndrome and a thickened, poorly functional bladder wall after early second-trimester decompression of fetal megacystis.

The management of bladder outlet obstruction in the fetus remains a debated issue. Recent data suggest the use of cystocentesis for the possible relief of the bladder neck obstruction or for the opening of posterior urethral valves. This case report adds further support for the use of cystocentesis and observation prior to the placement of an indwelling bladder shunt. The case further demonstrates that relief of the megacystis may not prevent prune belly syndrome, bladder dysfunction or renal damage. Finally, the case demonstrates that severe oligohydramnios in the second trimester need not invariably result in pulmonary hypoplasia.

Effects of hyperglycemia on mitochondrial morphology in the region of the anterior neuropore in the explanted rat embryo model: evidence for a modified Reid hypothesis as a mechanism for diabetic teratogenesis.

Congenital malformations are the leading cause of the increased perinatal mortality in the infants of insulin-dependent diabetic mothers. The mechanisms(s) of diabetic teratologic development has yet to be defined. Hyperglycemia is known to depress aerobic metabolism in many organisms and cell lines. Reid hypothesized that exposure to hyperglycemia could result in decreased mitochondrial biogenesis in embryonic cells. Should these cells suddenly be changed to an environment of lower glucose concentration, decreased energy capabilities would exist until sufficient numbers of mitochondria could be regenerated. Such cells may not be capable of meeting temporal-spatial requirements, thereby resulting in structural abnormalities. In our study the explanted rat embryo model demonstrated that in the head-fold region hyperglycemia produced morphologic alterations of mitochondria but no difference in the number of mitochondria per cell. Specifically, embryos cultured in euglycemia demonstrated orthodox mitochondrial configuration, whereas those cultured in hyperglycemia had mitochondria in a condensed configuration. These findings were reversible. A modification of Reid's original hypothesis may provide an explanation for the mechanism of diabetic teratologic development.

Hypomandibular faciocranial dysostosis: report of an affected sib and follow-up.

Hypomandibular faciocranial dysostosis is a condition heretofore described only in a single case. We report the birth of an affected sister along with follow-up information on the initial surviving patient. While a primary error in neural crest development was postulated in this syndrome, subsequently discovered anatomical abnormalities suggest a more complex pathogenesis.

Measurement of hypoxanthine and xanthine in late-gestation human amniotic fluid by reversed-phase high-performance liquid chromatography with photodiode-array detection.

A robust analytical method was developed for measurement of hypoxanthine and xanthine in late-gestation human amniotic fluid by reversed-phase high-performance liquid chromatography using diodearray detection. Purity of analyte peaks was confirmed via on-line analysis of peak spectra utilizing the purity parameter treatment of spectral data. Amniotic fluid obtained by amniocentesis was deproteinized by centrifugal ultrafiltration and chromatographed on an octadecylsilica column using isocratic elution with 1% (v/v) acetonitrile in 0.05 M ammonium dihydrogenphosphate pH 6.0; hypoxanthine and xanthine were resolved, but the hypoxanthine peak was not pure. Chromatography on a column of polar endcapped octadecylsilica, using similar mobile phase conditions, yielded spectrally pure peaks of hypoxanthine xanthine. Hypoxanthine and xanthine levels in amniotic fluid from fourteen patients, gestational age 34-39 weeks, ranged from 0.56 to 2.74 microM and 1.62 to 5.52 microM, respectively.

Acute coagulopathy in pregnancy.

Pregnancy alters both the fibrinolytic system and coagulation cascade. In addition, pregnancy presents unique triggering mechanisms for DIC. Management of DIC in pregnancy should include removal of the triggering mechanism, blood, and factor replacement. Inherited coagulation defects, while rarely resulting in bleeding diathesis in the pregnant patient, do require monitoring of maternal factor levels. Genetic counseling should be offered to all patients with inheritable coagulation disorders.

Emergent cesarean delivery in patients undergoing a trial of labor with a transverse lower-segment scar.

The American College of Obstetricians and Gynecologists recommends that women with previous low transverse cesarean sections who are candidates for a trial of labor should undergo such a trial only in a special environment. Such recommendations imply that these women are at greater risk than the general population for developing complications necessitating emergent operative intervention. A retrospective review of 5 years' experience reveals that in 1156 term pregnancies in which the patients were given a trial of labor, 18 (1.56%) patients underwent emergent abdominal delivery. This rate and the indications for the emergent operative delivery were not found to be different from those cited for patients without a uterine scar. The incidence of symptomatic uterine rupture was higher in patients scheduled for repeat cesarean section than for patients who underwent a trial of labor. Recommendations for a trial of labor might be reconsidered as they imply an increased risk for maternal and fetal morbidity and mortality that has yet to be demonstrated.

Breast stimulation in cycling women, pregnant women and a woman with induced lactation: pattern of release of oxytocin, prolactin and luteinizing hormone.

Levels of oxytocin (OT) and PRL were measured in plasma drawn before and during intermittent mechanical pump and tactile stimulation of the breast in five normal cycling women and 19 women in the third trimester of pregnancy. OT was significantly increased above baseline in response to breast stimulation in two of five cycling women and PRL increased in one of the two OT responders. In pregnant women, mean OT post nipple stimulation was significantly higher than pre nipple stimulation whereas PRL did not increase significantly. The response of OT to nipple stimulation occurred in 18 of 19 pregnant women compared to only two of five normal cycling women but the magnitude of the OT response in pregnant women was less than in cycling women or post-partum lactating women previously studied in this laboratory. In one non post-partum woman who induced lactation for the purpose of breast-feeding an adopted infant, OT and PRL were measured before and during mechanical pump and tactile stimulation before initiation of breast-feeding. OT increased during mechanical pump and tactile stimulation of the breast, as well as suckling, whereas PRL increased only in response to suckling. Levels of LH were measured in plasma every 20 min for 160 min at the following times: before initiation of breast-feeding, during induced lactation while breast-feeding, and 30 d after discontinuation of breast-feeding. Despite the development of oligomenorrhoea during the period of breast-feeding, levels of progesterone were not suppressed and LH was released in a normal pulsatile fashion.

Oxytocin and prolactin responses associated with nipple stimulation contraction stress tests.

Levels of oxytocin in plasma of pregnant women in the third trimester undergoing nipple stimulation-contraction stress tests were found, by parametric (paired t test) statistical analysis to be elevated. No differences in the plasma increases in oxytocin levels were found between those patients undergoing successful and those undergoing unsuccessful nipple stimulation-contraction stress tests. No significant differences in plasma levels of prolactin were associated with nipple stimulation in these patients.