Whole mount immunofluorescence analysis of placentas from normotensive versus preeclamptic pregnancies.

INTRODUCTION: Defects in placental angiogenesis and spiral artery remodeling have been proposed to play essential roles in the development of preeclampsia. However, the specific molecular mechanism(s) responsible for aberrant placental angiogenesis in preeclampsia are incompletely understood. The vascular endothelial growth factor receptors (VEGFR1, R2, R3) and STAT3 have critical functions in normal blood vessel development, but their potential roles in preeclampsia are currently unclear. In this study, we utilized a novel whole mount immunofluorescence (WMIF) method to compare expression of VEGFR1, R2, R3 and activated, phosphorylated STAT3 (pSTAT3) in placentas of preeclamptic (PE) versus normotensive (NT) pregnancies. METHODS: Placental biopsies collected from NT and PE pregnant women were fixed and stained with fluorochrome-conjugated antibodies to identify specific cell populations as follows: CD31 for blood vessel endothelial cells, cytokeratin-7 for trophoblast cells, and CD45 for immune cells. Expression of the VEGFRs and pSTAT3 were subsequently characterized by WMIF in conjunction with confocal microscopy. RESULTS: A total of 18 PE and 18 NT placentas were evaluated. No significant differences in the cell type-specific expression patterns or expression levels of VEGFR1, VEGFR2 or VEGFR3 were detected between NT and PE placentas. In contrast, statistically significant increases in pSTAT3 staining were detected in endothelial cells of PE placentas versus NT controls. DISCUSSION: Our study demonstrates that increased pSTAT3 expression in placental endothelial cells is associated with PE. We speculate that elevated pSTAT3 expression in the blood vessels of PE placentas may be due to aberrant angiogenesis, increased pro-inflammatory cytokine expression, and/or placental stress.

Maternal diet but not gestational weight gain predicts central adiposity accretion in utero among pregnant adolescents.

BACKGROUND: Modifiable risk factors during pregnancy, such as diet and weight gain, are associated with fetal birth weight but little is known about how these factors influence fetal fat acquisition in utero among pregnant adolescents. OBJECTIVE: To determine whether maternal pre-pregnancy BMI (ppBMI), gestational weight gain (GWG) and dietary intake during pregnancy influence fetal fat accretion in utero. METHODS: Longitudinal data were obtained from 121 pregnant adolescents enrolled in a study designed to identify determinants of maternal and fetal bone changes across gestation. Adolescents (ages 13-18 years) completed up to three study visits during early, mid- and late gestation. Maternal anthropometrics, 24 h dietary recalls and measures of fetal biometry were obtained at each visit. Fetal abdominal wall thickness (abdominal subcutaneous fat thickness, AbFat), a measure of fetal subcutaneous fat, was calculated by sonography at each visit. Statistical determinants of AbFat during late pregnancy were explored using simple and multiple regression. RESULTS: During late pregnancy (34.8±2.0 weeks; range 31.0-40.6 weeks of gestation), the median (inter-quartile range) fetal AbFat and GWG were 0.44 (0.39, 0.55) cm and 14.6 (9.5, 18.3) kg, respectively. After adjusting for infant birth weight, variables significantly associated with fetal AbFat included gestational age (P<0.0001, 95% confidence interval, CI: 0.01, 0.03), maternal race (P=0.029, 95% CI: -0.04, -0.002) and dietary intake of added sugar (P=0.025, 95% CI: 1.42e-6, 2.06e-5). Fetal AbFat had a significant positive quadratic relationship with total maternal dietary sugar intake such that both low and high extremes of sugar consumption were associated with significantly higher fetal AbFat. Birth weight was not significantly associated with maternal intake of added sugars. CONCLUSION: Extreme sugar intakes among pregnant adolescents may lead to increased accumulation of fetal abdominal fat with little net effect on birth weight. This finding suggests that increased sugar consumption during pregnancy promotes shifts in fetal body composition.

Fetal nuchal translucency thickness evaluation in the overweight and obese gravida.

OBJECTIVE: To evaluate the time required and failure rate for completion of nuchal translucency thickness (NT) measurements with increased maternal body mass index (BMI). METHODS: This was a retrospective review of ultrasound examinations for NT measurement in 11-14-week singleton pregnancies performed at a single site from 2004 to 2007. Women were stratified by prepregnancy BMI into normal weight (BMI, 18.5-24.9 kg/m(2)), overweight (BMI, 25.0-29.9 kg/m(2)) and obese Class I (BMI, 30.0-34.9 kg/m(2)), Class II (BMI, 35.0-39.9 kg/m(2)) and Class III (BMI >or= 40.0 kg/m(2)) groups. The failure rate, the time required for measurement, and the total study time in min were evaluated by BMI class for the first attempt and for all attempts at NT measurement. RESULTS: A total of 2508 women underwent attempted NT screening with complete data available on 1678 women (1707 examinations). The failure rate for NT screening varied significantly according to BMI (P < 0.001). At the first attempt, the median time for NT measurement varied significantly according to BMI (normal weight group, 9.7 (interquartile range (IQR) 4.4, 19.0) min; overweight group, 8.8 (4.0, 19.8) min; obese Class I, 9.6 (4.8, 20.4) min; Class II, 14.1 (5.0, 28.2) min; Class III, 12.3 (4.6, 22.7) min; P < 0.01), as did the total study time (normal group, 16.4 (10.1, 26.6) min; overweight group, 15.7 (9.8, 25.0) min, Class I, 17.3 (10.3, 29.2) min; Class II, 23.0 (12.2, 36.1) min; Class III, 18.7 (11.0, 30.8) min; P = 0.002). For all attempts also, the median time for NT measurement varied significantly according to BMI (normal weight group, 9.7 (IQR 4.4, 19.0) min; overweight group, 8.8 (4.0, 19.9) min; obese Class I, 9.6 (4.8, 21.0) min; Class II, 14.1 (5.0, 28.7) min; Class III, 12.3 (4.6, 22.5) min; P < 0.01), as did the total study time (normal weight group, 16.4 (10.2, 26.7) min; overweight group, 15.7 (9.8, 25.1) min; Class I, 17.6 (10.4, 29.9) min; Class II, 23.2 (12.0, 37.5) min; Class III, 18.7 (11.9, 31.9) min; P < 0.001). CONCLUSION: As maternal BMI increases, the time required to obtain NT measurements and the failure rate increase. Before the ultrasound examination, patients with a BMI over 30 should be counseled regarding the need for additional time and failure rates for first-trimester screening.

Fetal anatomic evaluation in the overweight and obese gravida.

OBJECTIVES: To evaluate the rate of completion of anatomic surveys of fetuses in overweight and obese gravid patients as compared with normal controls. METHODS: This was a retrospective review of anatomic ultrasound scans performed between 2004 and 2007. Women were stratified by prepregnancy body mass index (BMI) into normal weight (BMI, 18.5-24.9 kg/m(2)), overweight (BMI, 25.0-29.9 kg/m(2)) and obese Class I (BMI, 30.0-34.9 kg/m(2)), Class II (BMI, 35.0-39.9 kg/m(2)) and Class III (BMI >or= 40.0 kg/m(2)) groups. Rates of completion of basic and comprehensive scans, gestational age at completion and number of scans required were compared. RESULTS: For the 7140 patients included, completion rates for both the basic (normal weight, n = 2253 (79%); overweight, n = 1771 (76%); obese Class I, n = 767 (72%), Class II, n = 323 (61%) and Class III, n = 171 (49%)) and comprehensive (normal weight, n = 1234 (43%); overweight, n = 930 (40%); obese Class I, n = 404 (38%), Class II, n = 215 (41%) and Class III, n = 108 (31%)) surveys decreased significantly with increasing BMI (P < 0.00001). For surveys completed, the mean number of scans required was significantly higher for obese patients (basic: normal weight 1.3 vs. obese Class III 1.9; comprehensive: normal weight 1.7 vs. obese Class III, 2.2)(P < 0.00001). The overall completion rate improved at each gestational week, but was best between 20 and 23 weeks for obese patients. CONCLUSIONS: As maternal BMI increases, the rate of completion of anatomic surveys decreases and the number of scans required increases. Delaying the initial survey until 20 weeks' gestation may improve the capacity to complete the examination in a single visit. It should be noted that these results represent completion rates at a tertiary referral center, and therefore may not reflect community experience.

Sonographic birth-weight prediction in obese patients using the gestation-adjusted prediction method.

OBJECTIVES: Ultrasound birth-weight prediction may be more accurate if assessed at 34 to 36 + 6 weeks' gestation and extrapolated using the gestation-adjusted projection (GAP) method than if done at term. Because ultrasound is less accurate in women with elevated body mass index (BMI), we assessed the accuracy of GAP birth-weight prediction in obese as compared to non-obese women. METHODS: We performed a retrospective review of 1382 women with singleton pregnancies who had undergone fetal ultrasound examination at between 34 + 0 and 36 + 6 weeks, subclassified by pre-pregnancy BMI. Analysis of variance was used to compare predicted and actual birth weight. RESULTS: 1025 controls and 357 obese women were included. The obese women were divided by BMI: 159 in Class I (BMI, 30-34.9 kg/m(2)); 105 in Class II (BMI, 35-40 kg/m(2)) and 93 in Class III (BMI > 40 kg/m(2)). Mean systematic (percent) birth-weight prediction error was within 4% for all groups, with a 95% error range between - 5% and + 5%. The GAP method was able to predict actual birth weight within 20% for all groups in over 90% of cases. For all groups, the GAP method correctly excluded the presence of macrosomia with >or= 90% specificity. Negative likelihood ratios for the prediction of macrosomia were between 0.4 and 0.6 for all groups, regardless of obesity. CONCLUSIONS: The GAP method of birth-weight prediction using ultrasound measurement at 34 to 36 + 6 weeks predicts birth weight within 20% error in over 90% of cases, and is able to exclude macrosomia with over 90% accuracy regardless of maternal BMI.

Pregnancy and birth outcomes in women with facioscapulohumeral muscular dystrophy.

Obstetric risk in facioscapulohumeral muscular dystrophy (FSHD) is not known. We surveyed 38 women with FSHD reporting 105 gestations and 78 live births. Review of medical records showed that pregnancy outcomes were generally favorable. The rates for low birth weight and total operative deliveries were statistically higher than the national rates in the general population. Worsening of FSHD was reported in 24% of gestations and did not usually resolve after delivery.

Cross-correlation of fetal cardiac and somatic activity as an indicator of antenatal neural development.

OBJECTIVE: In this study, we wanted to model the emergence of coupling between fetal cardiac and somatic activity in normal and at-risk fetuses. STUDY DESIGN: One hundred six fetuses of uncomplicated pregnancies were longitudinally monitored at 20, 24, 28, 32, 36, and 38 weeks of gestation by using a fetal actocardiograph and computerized data collection. Twenty-six fetuses of complicated pregnancies were also included. Statistical time series analysis techniques were used to examine the relation between fetal movement and fetal heart rate. RESULTS: A linear increase was found in the magnitude of the cross-correlation function between fetal movement and fetal heart rate as gestation advanced, with coalescence around a peak lag of 5 seconds by 32 weeks. Fetuses that delivered before term evidenced accelerated fetal movement and fetal heart rate coupling, whereas fetuses affected by deleterious conditions showed a decline in developmental trajectory. CONCLUSIONS: The cross-correlation between fetal cardiac and somatic activity is an indicator of neuroregulation in human fetuses.

University hospital-based prenatal care decreases the rate of preterm delivery and costs, when compared to managed care.

OBJECTIVE: Our purpose was to compare the rate of recurrent preterm delivery and the combined costs of mother/infant care for patients with a history of preterm delivery cared for in an inner city hospital house staff (HS) clinic versus an inner city managed care organization (MCO). METHODS: A retrospective cohort study was conducted. The groups consisted of 96 patients with a history of preterm delivery who were cared for by the HS clinic and 164 patients cared for in a neighborhood MCO. All patients with a history of previous preterm delivery who delivered at the Johns Hopkins Hospital between 1 January 1994 and 31 December 1996 were included in the study. The groups were examined regarding source of prenatal care for the subsequent pregnancy (HS vs. MCO), baseline demographics, intensity of prenatal care, maternal and neonatal outcomes and total cost of the provision of care. RESULTS: There were no differences in baseline demographics between the groups. There was a higher rate of recurrent prematurity for patients cared for by the MCO (HS, 24% vs. MCO, 36%, p = 0.04). Mean total mother/infant costs were also higher in the MCO group ($13565) when compared to the HS group ($9082), (p = 0.02). CONCLUSION: While some MCOs may provide cost savings for some low-risk populations, our study demonstrates that this health-care delivery system resulted in greater total expenditures for patients with a history of preterm delivery.

Antenatal origins of individual differences in heart rate.

This study examines prenatal-to-postnatal stability in heart rate and variability from mid-gestation through the first year of life. Fetal heart rate data were collected from 52 healthy fetuses at 24, 30, and 36 weeks gestation, and again at 2 weeks and 12 months of age. Fetal heart rate measures were stable during gestation and positively associated with neonatal and infant measures. Maternal pulse rate and oxygen saturation were moderately associated with fetal heart rate. Together, fetal cardiac (heart rate and variability) and maternal physiologic measures (blood pressure and oxygen saturation) explained 40 and 48% of the variance in heart rate and variability, respectively, at 1 year of age. These common measures of individual differences in autonomic function are enduring characteristics that originate during fetal development.

Prediction of birth weight by ultrasound in the third trimester.

OBJECTIVE: To compare the accuracy of predicted birth weight by the gestation-adjusted projection method using ultrasonographic measurements obtained just before and at term. METHODS: The study group comprised patients with singleton pregnancies who underwent sonograms between 34.0 and 36.9 weeks' gestation (period 1) and at 37 weeks and beyond (period 2). The mean error in birth weight prediction, absolute birth weight error, and signed and absolute percent errors were compared with paired t tests. Thus, each patient served as her own control. RESULTS: The study included 138 patients undergoing 276 sonograms. The mean absolute error of the predicted birth weight was smaller for period 1 than for period 2 (197 +/- 167 g compared with 235 +/- 209 g, P =.019). The mean absolute percent error was 6.2 +/- 5.2% for period 1 compared with 7.4 +/- 6.3% for period 2 (P =.019). These same trends were observed when fetuses with suspected growth abnormalities were examined separately. Averaging data from both gestational periods did not improve the prediction of birth weight. CONCLUSION: Sonograms between 34.0 and 36. 9 weeks' gestation allow for more accurate prediction of birth weight than sonograms later in gestation. Though these differences are small and not clinically significant, this study indicates that serial sonograms in the late third trimester do not improve the ability to predict birth weight, even in abnormally grown fetuses. A single sonogram between 34 and 37 weeks' gestation is recommended for prediction of birth weight.

Fetal movement detection: comparison of the Toitu actograph with ultrasound from 20 weeks gestation.

OBJECTIVE: This study evaluates the validity of Doppler-detected fetal movement by a commercially available monitor and investigates whether characteristics of maternal body habitus and the intrauterine environment affect its performance. METHODS: Fetal movement was evaluated in normal pregnancies using both ultrasound visualization and a fetal actocardiograph (Toitu MT320; Tofa Medical Inc., Malvern, PA). Data were collected for 32 min on 34 fetuses stratified by gestational age (20-25 weeks; 28-32 weeks; 35-39 weeks). Fetal and maternal characteristics were recorded. Comparisons between ultrasound-detected trunk and limb movements and actograph records were conducted based both on 10-s time intervals and on detection of individual movements. RESULTS: Time-based comparisons indicated agreement between ultrasound and actograph 94.7% of the time; this association rose to 98% when movements of less than 1 s duration were excluded. Individual movements observed on ultrasound were detected by the actograph 91% of the time, and 97% of the time when brief, isolated movements were excluded. The overall kappa value for agreement was 0.88. The actograph was reliable in detecting periods of quiescence as well as activity. These findings did not vary by gestational age. The number of movements detected by the actograph, but not the single-transducer ultrasound, significantly increased over gestation. Maternal age, parity, weight, height, or body mass index were not consistently associated with actograph validity. Characteristics of the uterine environment, including placenta location, fetal presentation, and amniotic fluid volume also did not affect results. CONCLUSIONS: The Toitu actograph accurately detects fetal movement and quiescence from as early as 20 weeks gestation and has utility in both clinical and research settings. Actographs are most useful for providing objective and quantifiable measures of fetal activity level, including number and duration of movements, while visualization through ultrasound is necessary for studies of movement quality, source, or mechanics.

Consent documents, reproductive issues, and the inclusion of women in clinical trials.

PURPOSE: To describe language used in consent documents at one academic medical center to inform women participating in studies of potential reproductive and fetal risks. METHOD: The authors reviewed consent document language describing reproductive and fetal risks in 114 approved protocols. Protocols were identified as being of high, low, or unknown risk based upon FDA drug-risk and radiation-risk categories. RESULTS: Although most consent documents advised women against participating for one or more pregnancy-related reasons, specific information about reproductive or fetal risks was included in fewer consent documents: 8 (73%) of the high-risk studies, 12 (40%) of the low-to-moderate-risk studies, and 29 (40%) of the unknown-risk studies. CONCLUSIONS: Investigators often omit fetal risk information from consent documents. Full disclosure of reproductive and fetal risks in consent documents and discussions can be taught and modeled during the research training period. The authors present a template with language that can be used in consent documents and recommend ongoing discussion of reproductive and fetal risks with women subjects throughout the study period.

[Protein E 98-113 sequence is a fusion site of tick-borne encephalitis virus with cellular membrane]. / Posledovatel'nost' 98-113 belka E virusa kleshchevogo éntsefalita iavliaetsia uchastkom sliianiia virusa s kletochnoi membranoi.

The synthetic peptide with the conservative 98-113 sequence of protein E of tick-borne encephalitis virus was studied in order to elucidate its role in the functioning of flaviviruses. The peptide was shown to inhibit the in vitro infection of macrophages with the virus. An antibody that specifically binds this peptide was found among the set of monoclonal antibodies produced against protein E. This antibody was found to prevent penetration of the virus into liposomes. A correlation was found between our results and data on the spatial structure of protein E and its interspecies homology. The protein E 98-113 sequence of the tick-borne encephalitis virus was found to be the fusion site of the viral envelope with a cellular membrane.

[Preparation and properties of monoclonal antibodies to tick-borne encephalitis viral nonstructural proteins]. / Poluchenie i svoistva monoklonal'nykh antitel k nestrukturnym belkam virusa kleshchevogo entsefalita.

Hybridomas secreting monoclonal antibodies (Mab) to tick-borne encephalitis (TBE) virus are obtained. Immunodiffusion showed that 3 Mabs to TBE protein NS3 belong to class IgM and the rest to IgG1. Mabs to TBE protein NS1 were tested in hemagglutination inhibition, complement fixation, neutralization, and protection tests. Only 1 hybridoma produced Mab specific for protein NS1 of TBE strain Sofyin, the rest reacted with the common antigenic determinants of nonstructural TBE complex.

Fetal neurobehavioral development: associations with socioeconomic class and fetal sex.

This longitudinal study investigated neurobehavioral development in the human fetus from 24 to 36 weeks gestation. Subject (N=103) were stratified by socioeconomic class. Fetal data were collected for 50 min at three intervals, and included measures of heart rate, movement, and biobehavioral patterns. Repeated measures analysis of variance by fetal sex and maternal socioeconomic status was used to detect maturation effects and group differences. With advancing gestation, fetuses exhibited reduced heart rate, increased heart rate variability and coupling between movement and heart rate, increased movement vigor, and more biobehavioral concordance. Male fetuses displayed higher heart rate variability throughout gestation and somewhat earlier emergence of biobehavioral organization than females. Fetuses of women of lower socioeconomic status had reduced heart rate variability, moved less often and with less vigor, showed less coupling between movement and heart rate, and had fewer episodes of synchronous quiescence/activity. Results are discussed in terms of development of the central nervous system.

Managed care does not lower costs but may result in poorer outcomes for patients with gestational diabetes.

OBJECTIVE: Our purpose was to compare the costs of prenatal care and subsequent maternal and neonatal outcomes in patients with gestational diabetes cared for in an inner-city university hospital house staff clinic versus an inner-city managed care organization. STUDY DESIGN: A retrospective cohort study was conducted. The groups consisted of 115 patients with gestational diabetes who were cared for in a house staff clinic and a demographically similar group of 85 patients cared for in a neighborhood managed care organization. The groups were examined regarding baseline demographics, intensity of prenatal care, maternal and neonatal outcomes, and total cost of the provision of care. RESULTS: There was no difference between groups in the total cost of maternal-infant care. A larger percentage of patients in the house staff group saw the physician frequently. In contrast, patients cared for in the managed care organization underwent more tests of fetal well-being. There was a greater rate of neonatal macrosomia in the managed care organization group compared with the house staff group. CONCLUSIONS: Managed care does not decrease the cost of caring for patients with gestational diabetes but does lead to a greater rate of neonatal macrosomia, which may reflect poorer glucose control.

[A comparative analysis of proteins from persistent and antigen-defective strains of tick-borne encephalitis virus]. / Sravnitel'nyi analiz belkov persistiruiushchikh i antigenno-defektnykhshtammov virusa kleshchevogo éntsefalita.

Structural (E) and three nonstructural (NS1, NS3, and NS5) proteins of persistent and antigen-defective strains of tick-borne encephalitis virus are compared by immunoblotting with monoclonal antibodies to the corresponding proteins of strain Sofyin. Appreciable phenotypical differences were revealed between antigen-defective strains, but no immunological modifications as concerns the studied antigenic structures. The size of the reference NS3 protein differed from that of the persistent virus strains. Similar proteins detected in the antigenic preparations of strain Zausaev may have notable modifications.

A prospective randomized trial of two solutions for intrapartum amnioinfusion: effects on fetal electrolytes, osmolality, and acid-base status.

OBJECTIVE: Our purpose was to compare the effects of intrapartum amnioinfusion with normal saline solution versus lactated Ringer's solution plus physiologic glucose on neonatal electrolytes and acid-base balance. STUDY DESIGN: Patients undergoing amnioinfusion for obstetric indications were randomized to receive normal saline solution or lactated Ringer's solution plus physiologic glucose at standardized amnioinfusion rates. Data were collected prospectively on maternal demographics, course of labor, and maternal and neonatal outcome. Arterial cord blood was obtained for analysis of electrolytes, glucose, osmolality, lactic acid, and blood gases. Control subjects with normal fetal heart rate patterns, and clear amniotic fluid not receiving amnioinfusion were studied concurrently. RESULTS: Data were collected on 59 patients (21 normal saline solution, 18 lactated Ringer's solution plus physiologic glucose, and 20 controls). Maternal demographics, course of labor, and neonatal outcome were similar in all three groups. Cesarean sections were performed more often in the amnioinfusion groups (33.3% for normal saline solution, 38.9% for lactated Ringer's solution plus physiologic glucose) than in the control group (5.0%), p < 0.05. Cord arterial electrolytes, glucose, osmolality, lactic acid, and blood gases were not altered by amnioinfusion with either solution. CONCLUSION: Intrapartum amnioinfusion with normal saline solution or lactated Ringer's solution plus physiologic glucose has no effect on neonatal electrolytes or acid-base balance.

Differentiating lymphocytic adenohypophysitis from pituitary adenoma in the peripartum patient.

Lymphocytic adenohypophysitis (LAH) is an autoimmune disorder of the pituitary gland with a predilection for the peripartum period and often mimics a pituitary adenoma. We sought to define the clinical, endocrinologic and radiographic characteristics differentiating peripartum LAH from pituitary adenoma to enable the use of noninvasive diagnosis and appropriate therapy. From published reports and our own case, the clinical histories and laboratory and radiographic studies of 45 patients fulfilling the diagnosis of peripartum LAH were reviewed. History of infertility or menstrual irregularity, symptomatology, endocrinologic evaluation, diagnostic imaging and associated medical conditions were analyzed. For comparison, 806 patients with pituitary adenoma and pregnancy from published series were evaluated. The spontaneous pregnancy rate in pituitary adenoma patients was 2.4% vs. 100% in LAH patients. Visual disturbances and headaches were significantly more frequent in patients with LAH. Prolactin levels were significantly lower in patients with LAH than in those with pituitary adenomas (34.6 +/- 46.3 [SD] vs. 393.0 +/- 300.4, P < .0001). Abnormalities in thyroid and/or adrenal function were also more common in patients with LAH (57.5% vs. 2.5%, P < .001). There were no distinguishing characteristics on radiographic studies. History and endocrinologic evaluation can differentiate between LAH and pituitary adenoma in the peripartum patient.