Prospective multicenter study of ultrasound-based measurements of fetal head station and position throughout labor.

OBJECTIVES: To assess the relationship between fetal head position and head station during labor, as measured using an ultrasound-based system, and the occurrence of occiput posterior (OP) position at delivery. METHODS: This was an international prospective observational study including women who delivered between January 2009 and September 2013 in four centers: one in Brooklyn, NY, USA; one in Haifa, Israel; and two in Paris, France. We used an ultrasound-based system (LaborPro) to monitor fetal head station and position non-invasively throughout labor. We collected data on demographics, labor parameters and outcome. RESULTS: A total of 595 women were included. In 563 (94.6%) women, fetal head position at delivery was occiput anterior (OA), in 31 (5.2%) it was OP and in one (0.2%) it was occiput transverse. In 89% of pregnancies with intrapartum OP when fetal head station was above -2, the head position turned to OA at delivery; the equivalent figures were 74% and 63% OA at delivery when intrapartum OP was diagnosed at head stations of -2 to < 0, and 0 and below, respectively. Cesarean delivery was performed in 35% of pregnancies with fetal head in OP position at delivery, as opposed to 10% of those with non-OP position at delivery. On retrospective analysis, all deliveries in OP were already in OP at station -2 and below. CONCLUSIONS: In this first assessment of fetal head position at delivery according to fetal head position at various station levels, our data show that 100% of OP positions at delivery were already in OP position at station -2 and below. We did not observe rotation from a non-OP to an OP position from station -2 and below. Nearly two-thirds of fetuses in OP at station 0 and below will rotate to an OA position for delivery.

Placental protein 13 and decidual zones of necrosis: an immunologic diversion that may be linked to preeclampsia.

We evaluated the role of placental protein 13 (PP13; galectin 13) in the process of trophoblast invasion and decidual necrosis. Immunohistochemical analysis for PP13, immune cells, human placental lactogen, cytokeratin, and apoptosis markers was performed on 20 elective pregnancy termination specimens between 6 and 15 weeks of gestation. Placental protein 13 was localized to syncytiotrophoblasts in the chorionic villi and to occasional multinucleated luminal trophoblasts within converted decidual spiral arterioles. Cytotrophoblasts, anchoring trophoblasts, and invasive trophoblasts did not stain for PP13. Extracellular PP13 aggregates were found around decidual veins associated with T-cell-, neutrophil- and macrophage-containing decidual zones of necrosis (ZONEs). We hypothesize that PP13 is secreted into the intervillus space, drains through the decidua basalis veins, and forms perivenous PP13 aggregates which attract and activate maternal immune cells. Thus, syncytiotrophoblast-derived PP13 may create a ZONE that facilitates trophoblast invasion and conversion of the maternal spiral arterioles.

Cervical varices complicated by thrombosis in pregnancy.

We present a case of a 36-year-old pregnant woman with varicose veins of the uterine cervix diagnosed during the second trimester. The cervical tissue was completely replaced with dilated venous plexus. Therapeutic decisions were further complicated by the presence of blood clots within the dilated vessels. Pregnancy proceeded uneventfully, no bleeding occurred and the patient underwent a planned Cesarean delivery. This is the first reported case in the medical literature where the rare phenomenon of cervical varices was accompanied by thrombosis.

Placenta-bound and body fluid PP13 and its mRNA in normal pregnancy compared to preeclampsia, HELLP and preterm delivery.

OBJECTIVES: To compare the distribution of placental protein 13 (PP13) in fetal and maternal blood and amnionic fluid and to correlate it with PP13 protein and mRNA in the placenta. METHODS: Umbilical arterial serum, amnionic fluid, maternal venous serum and placental tissues were collected from normal outcome pregnancies (N = 63) (GA>37), early onset preeclampsia (PE) (N = 12, GA: 26-33), and HELLP syndrome (N = 5, GA: 27-29). Because PE and HELLP cases delivered preterm, cases of preterm delivery (PTD) (N = 6, GA: 31-36) served as additional control. PP13 was determined by ELISA, Western blot, and immunohistochemistry. PP13 mRNA was measured by PCR (RT-PCR). Continuous parameters were compared by t-test, P < 0.05 was considered significant. RESULTS: In women with normal pregnancy outcome significantly higher PP13 levels were found in maternal serum compared to amnionic fluid and negligible amount was found in fetal serum. A similar pattern was identified in cases of PTD with concentrations similar to term control. In PE and HELLP cases PP13 levels in amnionic fluid level were more than twice compared to maternal serum (P < 0.001). Umbilical cord level was negligible in PE but high in HELLP corresponding to the much higher level of PP13 in this patient group compared to all others. In the placenta PP13 level in term controls was higher compared to PTD. In PE and HELLP (similar early delivery time as PTD) the level was significantly higher (P < 0.01) compared to PTD or term controls. PP13 mRNA levels in term control and PTD were similar while PP13 mRNA levels in PE and HELLP placentas were significantly lower compared to term controls or PTD or the two combined. Syncytiotrophoblast labeling appeared stronger in PE and HELLP compared to term controls and PTD. CONCLUSIONS: In all cases but HELLP, PP13 in fetal blood is very low indicating that routing of PP13 to fetal blood is limited and that the fetus is unlikely to generate PP13. PP13 mRNA is lower in the third trimester at the time of disease while protein level accumulates and become higher creating an unparallel change in the level of the mRNA and the corresponding protein.

Placental and trophoblastic in vitro models to study preventive and therapeutic agents for preeclampsia.

In the field of preeclampsia, enormous efforts are ongoing to identify biomarkers predicting the syndrome already in the first trimester of pregnancy. At the same time, there is the need for in vitro models to test such biomarkers prior to their use in clinical trials. In addition, in vitro models may accelerate the development and evaluation of the benefit of any putative therapeutics. Therefore, in vitro systems have been established to evaluate the release of biomarkers and measure the effect of putative therapeutics using placental villous explants as well as the choriocarcinoma cell line BeWo. For explants, a cryogenic method to freeze, transport and thaw villous explants was developed to use such tissues for a multi-site tissue culture evaluation. Here we focus on three out of many in vitro models that have been established for human placental trophoblast. (1) Choriocarcinoma cell lines such as BeWo, Jeg-3 and Jar cells (2) isolated primary trophoblast cells, and (2) villous explants from normal placentas delivered at term. Cell lines were used to assess the effect of differentiation and fusion on the expression and release of a preeclampsia marker (placental protein 13; PP13) and beta-hCG. Moreover, cell lines were used to study the effect of putative preeclampsia therapeutics such as vitamins C and E, heparin and aspirin on marker release and viability. Cryopreservation of villous explants enabled shipment to a remote laboratory and testing of parameters in different countries using explants from one and the same placenta. Recently published data make it tempting to speculate that the choriocarcinoma cell line BeWo as well as fresh and cryogenically stored placental villous explants may well serve as in vitro models to study preventive and therapeutic agents in the field of preeclampsia.

Cryogenic and low temperature preservation of human placental villous explants - a new way to explore drugs in pregnancy disorders.

BACKGROUND: A major handicap in cell culture studies using human tissues is the insufficient availability of fresh material on site. A method was developed for cryogenic storage and low temperature preservation of human placental villous explants, facilitating multi-site distribution for functional studies. METHODS: Explants from term placentas were incubated with cryoprotectant agents (dimethyl-sulfoxide (DMSO), ethylene glycol, propanediol or Aedesta), frozen in liquid nitrogen, thawed and then cultured in-vitro. Viability was assessed by comparing frozen and thawed explants with non-frozen controls for morphological changes, lactate dehydrogenase (LDH) release, placenta protein 13 (PP13) secretion, and PCNA Western blotting. Functional studies determined the effect of oxygen and magnesium on explant viability. RESULTS: Cryoprotection by 3 M DMSO best maintained explants' viability, morphological integrity and PP13 release after freezing and thawing from liquid nitrogen. The effect of oxygen and magnesium was used to test the functional viability of cultured explants, after freezing in liquid nitrogen and transfer to dry ice for 1-5 days on site or for shipment to a remote lab. The tested parameters were similar between controls and cryogenically treated explants in the remote lab and the lab of origin, demonstrating the possibility of cryostoring explants for functional studies. CONCLUSION: Cryogenically stored placental villous explants shipped frozen can serve as a useful tool for comparative functional studies of placental villous tissues. The results of this pilot study also open the way for multi-site studies associated with drug tailoring for pregnancy disorders.

Association between ultrasound-based assessment of fetal head station and clinically assessed cervical dilatation.

OBJECTIVES: To describe the association between ultrasound-based determination of fetal head station and clinical assessment of cervical dilatation during active labor. METHODS: From 427 women with singleton uncomplicated term pregnancies we obtained, during the active phase of labor, 907 pairs of measurements. Fetal head station and position were determined using the LaborPro system, based on position tracking and ultrasound imaging technology, and degree of cervical dilatation was determined by digital vaginal examination. The association between them was analyzed. RESULTS: The overall correlation between cervical dilatation and fetal head station was 0.64 (P < 0.001). Complete dilatation was observed in 78% of women with fetal head engagement, and in all women with a fetal head station of + 1.5 or more. CONCLUSIONS: There is good association between non-invasive ultrasound-based determination of fetal head station and clinically assessed cervical dilatation.

Pregenesys pre-eclampsia markers consensus meeting: What do we require from markers, risk assessment and model systems to tailor preventive strategies?

The Pregenesys Consensus Meeting held in Cambridge on 13 July 2009 was organized by the Pregenesys Consortium to review and critically discuss current knowledge regarding early markers of preeclampsia, to identify priorities and opportunities for future research, to consider issues that may need to be addressed in future recommendations and to highlight key issues in cost effectiveness and national policies concerning prediction and early screening for the risk of developing preeclampsia. This report summarizes the outcome of the Consensus Meeting and draws attention to issues for further investigation with specific regard to single versus multiple markers, early versus late risk identification, and the long-term effects on both maternal and perinatal health and the need to include these in any future cost-benefit assessment.

Effects of calcium, magnesium, low-dose aspirin and low-molecular-weight heparin on the release of PP13 from placental explants.

BACKGROUND: Preeclampsia is one of the leading causes for maternal and fetal morbidity. Attempts to prevent preeclampsia have already been made using low-dose aspirin, low-molecular-weight heparin (LMWH), and calcium supplementation. Magnesium sulphate is used at the time of disease to prevent eclampsia. Here we investigated the effect of these agents on PP13 release from placental explants. METHODS: Placentas harvested after C-section of term or preterm control and preeclampsia cases or first trimester terminations were used to obtain explants. Explants were incubated for 24h with/without respective agents, harvested, weighed and subjected to PP13 determination in the culture medium and the explant. LDH was used to determine viability. Dose response curves were obtained for each drug. P < 0.05 was considered significant. RESULTS: Exposure to magnesium (0.7-7g/day) slightly decreased PP13 release from controls, and slightly increased it in preeclampsia and first trimester termination. Calcium (0. 3-6g/day) showed a tendency to decrease the release in control and preeclampsia, whereas in first trimester release was increased in a bell-shaped manner. Aspirin (0-250 mg/day) tended to decrease the release in controls but increased it in a bell-shaped manner in first trimester and preeclampsia. LMWH showed no effect from 0 to 80 mg/day in controls but tended to decrease PP13 release in preeclampsia and first trimester. CONCLUSION: This data might point to a beneficial effect of aspirin and calcium supplementation in the first trimester of pregnancy and aspirin at the time of disease, although the interaction with the maternal system still needs to be elucidated.

Placental protein 13 as an early marker for pre-eclampsia: a prospective longitudinal study.

OBJECTIVE: To assess the value of placental protein 13 (PP13) as an early marker of pre-eclampsia. DESIGN: Sequential blood samples were obtained from women with singleton viable pregnancies at 6-10, 16-20 and 24-28 weeks of gestation. Samples were tested for PP13 using a solid-phase sandwich enzyme-linked immunosorbent assay. Levels were expressed as multiples of the medians (MoM) of the unaffected population. The slope or rate of change in PP13 concentration per week of gestation was also calculated. SETTING: Thirty-five prenatal care community clinics. SAMPLE: In total, 1,366 women were recruited, and subsequently, 20 were diagnosed with pre-eclampsia, 41 with gestational hypertension and 1,178 were unaffected. MAIN OUTCOME MEASURES: Sensitivity and specificity of screening with PP13 at each gestational period and of PP13 level combined with the slope of PP13 between two testing periods. RESULTS: At 6-10 gestational weeks, PP13 levels were significantly lower among the pre-eclampsia group with a median 0.28 MoM (95% CI 0.15-0.39, P < 0.004). Using a cutoff of 0.40 MoM, the sensitivity was 80%, false-positive rate (FPR) was 20% and odds ratio was 16.0 (95% CI 5.3-48.4). Combining MoM of 6-10 weeks and slope between 6-10 and 16-20 weeks, the sensitivity was 78%, the FPR was 6% and odds ratio was 55.5 (95% CI 18.2-169.2). The gestational hypertension group was not different from the normal group. CONCLUSIONS: PP13 in the first trimester alone or in combination with the slope between the first and the second trimesters may be a promising marker for assessing the risk of pre-eclampsia.

Determination of the solubility and size distribution of radioactive aerosols in the uranium processing plant at NRCN.

Inhalation is the main route of internal exposure to radioactive aerosols in the nuclear industry. To assess the radiation dose from the intake of these aerosols, it is necessary to know their physical (aerodynamic diameter distribution) and chemical (dissolution rate in extracellular lung fluid) characteristics. Air samples were taken from the uranium processing plant at the Nuclear Research Center, Negev. Measurements of aerodynamic diameter distribution using a cascade impactor indicated an average activity median aerodynamic diameter value close to 5 microm, in accordance with the recent recommended values of International Commission on Radiological Protection (ICRP) model. Solubility profiles of these aerosols were determined by performing in vitro solubility tests over 100 d in a simultant solution of the extracellular fluid. The tests indicated that the uranium aerosols should be assigned to an absorption between Types M and S (as defined by the ICRP Publication 66 model).

Normalisation of spot urine samples to 24-h collection for assessment of exposure to uranium.

For dose assessment of workers at Nuclear Research Center Negev exposed to natural uranium, spot urine samples are analysed and the results are normalised to 24-h urine excretion based on 'standard' man urine volume of 1.6 l d(-1). In the present work, the urine volume, uranium level and creatinine concentration were determined in two or three 24-h urine collections from 133 male workers (319 samples) and 33 female workers (88 samples). Three volunteers provided urine spot samples from each voiding during a 24-h period and a good correlation was found between the relative level of creatinine and uranium in spot samples collected from the same individual. The results show that normalisation of uranium concentration to creatinine in a spot sample represents the 24-h content of uranium better than normalisation to the standard volume and may be used to reduce the uncertainty of dose assessment based on spot samples.

An improved method for Organic Bound Tritium (OBT) determination in urine samples.

The Organic Bound Tritium (OBT) amount in the body may induce changes in the evaluation of the internal exposure to tritium, due to its different retention time relative to HTO. OBT measurements for urine are not performed routinely, mainly because of the lengthy work needed in preparation of the samples, when using the standard oxygen combustion method. A simpler and more rapid method based on liquid scintillation counting (LSC) was employed, and an evaluation was performed to check its suitability for urine samples. The principle of the proposed method is based on subtracting the LSC counts of the water phase sample (HTO) from the total activity of the urine sample (OBT+HTO). A good correlation was found between the results obtained when applying the proposed direct method and the Gold standard method. The minimum detectable activity when using this method was determined.

Three-dimensional power Doppler in the evaluation of painful leiomyomas and focal uterine thickening in pregnancy.

OBJECTIVE: To determine the usefulness of 3-dimensional (3D) power Doppler ultrasound in distinguishing painful leiomyomas from focal myometrial contractions or nonpainful leiomyomas in pregnancy. METHODS: A 2D section of the area of interest in the uterine wall was first obtained in 29 patients to determine whether the pain originated from a myoma or a uterine contraction. Then, volume acquisition was activated and Doppler indices (vascularization index, flow index, and vascularization-flow index) were calculated for thickened and normal uterine wall. RESULTS: Of 15 patients found to have uterine myomas, 11 had multiple tumors and 4 had single tumors. In the remaining 14 patients the painful uterine thickening proved to be a focal contraction. Painful segments of uterine thickening were associated with lower Doppler indices. Painful myomas were found to have significantly lower indices than normal uterine wall (P=0.008, 0.03, and 0.01 for VI, FI, and VFI, respectively, vs. 0.001, 0.003, and 0.01). However, the differences in indices between nonpainful myomas and uterine wall on the one hand, and nonpainful myomas and focal uterine contractions on the other, were not statistically significant. CONCLUSION: Three-dimensional power Doppler ultrasound may be a sensitive and reliable tool for distinguishing painful uterine myomas from focal myometrial contractions and nonpainful myomas.

Placental protein 13 (PP-13): effects on cultured trophoblasts, and its detection in human body fluids in normal and pathological pregnancies.

Placental tissue protein 13 (PP-13), one of the 56 known placental proteins identified till today, was purified from placentas obtained from women at delivery, and used to evoke antibodies against it. The purified PP-13 was lysed to peptides, which were sequenced, leading to the full-length cDNA sequencing and its expression in Escherichia coli. Sequence analysis in databases showed homology to the galectin family. Of the various antibody preparations developed, a pair of monoclonal antibodies (MAbs) coupled to the recombinant PP-13 (PP-13-R) was used for the immunodetection of PP-13 in pregnant women's serum with the solid-phase ELISA format. With a dynamic range of 25-500 pg/mL with no background in non-pregnant women's serum and men's serum, the ELISA test was suitable for the detection of PP-13 in the 1st, 2nd, and 3rd trimesters. PP-13 levels slowly increase during pregnancy. In the 1st trimester, lower than normal PP-13 levels were found in fetal growth restriction (IUGR), preeclampsia (PE), and particularly in early PE (<34 weeks of gestation). In the 2nd and 3rd trimesters, higher than normal concentrations were found in PE, IUGR and in preterm delivery (PTD). Application of PP-13 to cultured trophoblasts elicited depolarization carried by calcium ions, followed by liberation of linoleic and arachidonic acids from the trophoblast membrane, and a subsequent elevation of prostacyclin and thromboxane. These effects were negligible when PP-13 derived from the placentas of patients with IUGR, PE or PTD was used. The results are discussed in view of the potential utilization of PP-13 for early serum screening to assess the risk to develop placental insufficiency, coupled to a differential analysis of the various pathologies by analyzing cultured trophoblasts.

Obstetricians' opinions regarding patient choice in cesarean delivery.

OBJECTIVE: To survey the opinions of Israeli obstetricians regarding their position on patient choice cesarean delivery. METHODS: Short anonymous questionnaires were sent by mail. The questionnaire included basic demographic data followed by three short case presentations of patients requesting cesarean delivery without a medical indication; the respondents were asked if they would consent to the patients' request. Respondents were then asked about their personal preferences on the mode of delivery, their attitude towards women's right to choose cesarean delivery, and whether obstetricians should inform their patients of this right. RESULTS: Of the 650 questionnaires sent, 257 were returned. Most of the respondents were specialists, aged 35 years or older, and worked in hospitals. Seventy-five percent were male, and 27% had a teaching academic degree. The consent rate for patient choice cesarean delivery in the three case presentations ranged from 40% to 79%. Only 9% of the respondents said they would prefer cesarean delivery for themselves (if female) or for their partners. Forty-five percent supported women's right to choose cesarean delivery, and half of them stated that obstetricians should inform their patients of this right. CONCLUSION: Although the vast majority (91%) of the Israeli respondents personally prefer vaginal delivery, almost half of them support women's autonomy to choose cesarean delivery. Consequently, approximately 50% of the respondents were willing to perform cesarean delivery on request because of their support of women's autonomy, despite the fact that they believe that vaginal delivery is a better option.

Ultrasound evaluation of the fetal skull base throughout pregnancy.

OBJECTIVE: To construct nomograms for cranial base parameters of normal fetuses. STUDY DESIGN: A cross-sectional study of 386 normal singleton fetuses at 14-40 weeks' gestation. Measurements of the length and width of the sphenoid bone anteriorly and the otic cartilage posteriorly and of the angles between the ridges were obtained. RESULTS: Nomograms of the length and width of sphenoid ridge and otic cartilage, and of the angles in the cranial fossae are presented. A linear growth function was observed between cranial base measurements (sphenoid ridge length, otic cartilage length, cranial base angles) and gestational age, femur length and biparietal diameter. CONCLUSION: Measurements of the length of cranial base ridges and the angles between them can be obtained easily in the second and third trimesters and might prove useful in the evaluation of pregnancies at high risk for associated fetal abnormalities.

A new guidance system for freehand, obstetric ultrasound-guided procedures.

OBJECTIVE: To assess the performance of the UltraGuide 1000 system, and to compare ultrasound-guided freehand mid-trimester amniocentesis with and without the new guidance system. METHODS: One hundred and sixty-nine women referred for mid-trimester genetic amniocentesis were divided into two groups: a control group of 99 women who underwent the procedure by the freehand technique with scored needles and 70 patients who had the procedure carried out with the aid of a guidance system (UltraGuide 1000) with non-scored needles. The procedures were compared for duration, number of punctures and repositionings of the needle, the visibility of the needle during the puncture and the number of bloody taps. RESULTS: The study group had significantly lower rates of reinsertion (none vs. 7.1%), repositioning (7.1% vs. 17.7%), bloody taps (none vs. 6.1%), touching the fetus (5.7% vs. 22.2%) and prolonged duration of the procedure (4.3% vs. 14.4%) compared with the control group. There was one fetal loss in the control group. Non-visibility of the needle before reaching the amniotic sac occurred in 18.6% of cases in the study group and in 38.4% of cases in the control group. CONCLUSIONS: The new guidance system combines the benefits of an attached guide with the flexibility of the 'freehand' technique. Use of the new guidance system for mid-trimester genetic amniocentesis increases needle visibility and lowers the incidence of common complications.

Cardiac function in fetuses with intracardiac echogenic foci.

OBJECTIVE: To evaluate cardiac dimensions and function in euploid fetuses with intracardiac echogenic foci. STUDY DESIGN: Forty-eight fetuses with a single cardiac echogenic focus situated in the left ventricle had echocardiography performed at 22-24 weeks of gestation. Fifty normal fetuses at 22-24 weeks' gestation served as controls. Two-dimensional and M-mode directed fetal echocardiography were used to exclude cardiac anomalies and measure right and left ventricular free walls and interventricular septal thickness and ventricular systolic and diastolic dimensions. Cardiac size was expressed as a ratio of ventricular wall thickness/biparietal diameter, and cardiac function was expressed as ventricular shortening fraction. Doppler fetal echocardiography measurements included pulmonary and aortic maximum systolic velocities and time to peak velocities as indices of ventricular systolic function, and the ratio between early ventricular filling (E-wave) and active atrial filling (A-wave) peak velocities at the level of the atrioventricular valves as an index of ventricular diastolic function. RESULTS: Early ventricular filling/active atrial filling peak velocity ratios were significantly lower in fetuses with intracardiac echogenic foci than in control fetuses. In the mitral valve the ratio was 0.37 +/- 0.14 (0.039) (mean +/- SD (95% confidence interval for difference between the means)) vs. 0.59 +/- 0.19 (0.052) and in the tricuspid valve it was 0.42 +/- 0.16 (0.045) vs. 0.62 +/- 0.21 (0.058). No significant differences were found in cardiac dimensions, ventricular shortening fraction and Doppler systolic indices. CONCLUSION: Euploid fetuses with intracardiac echogenic foci show low E/A ratio values in midtrimester echocardiography. This finding might indicate cardiac diastolic dysfunction.

Candida sepsis following transcervical chorionic villi sampling.

BACKGROUND: The use of invasive devices and broad spectrum antibiotics has increased the rate of candidal superinfections. Candida sepsis associated with pregnancy is rare. Candida sepsis following chorionic villi sampling (CVS) has never been reported. CASE: A 31 -year-old pregnant woman presented with signs of sepsis one day after undergoing transcervical CVS. Blood culture and curettage material yielded C. albicans. She was treated with 400 mg of fluconazole daily for 4 weeks and completely recovered. CONCLUSION: Candida sepsis should be considered in the differential diagnosis of sepsis following CVS.