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We assessed whether the earlier described 'thin-fat phenotype' is present in Surinam South Asian babies of the fourth to fifth generation after migration from India. In this observational study we collected data from 39 South Asian term neonates and their mothers in Paramaribo, Surinam. We compared the following data with data from an earlier study in Southampton, UK (338 neonates) and in Pune, India (631 neonates): maternal body mass index, neonatal weight, length, head, mid-upper arm and abdominal circumferences and subscapular skinfold thickness. The mothers in Paramaribo were older than the Southampton mothers; their body mass index was comparable. Mean birth weight was 3159 g (Southampton: 3494 g; Pune: 2666 g). Compared with Southampton babies, the Paramaribo babies were smaller in nearly all body measurements, the smallest being abdominal circumference at the umbilicus level (s.d. score: -1.62; 95% confidence interval (CI): -2.07 to -1.16) and mid-upper arm circumference (s.d. score: -1.08; 95% CI: -1.46 to -0.69). In contrast, subscapular skinfold thickness was similar (s.d. score: +0.08; 95% CI: -0.24 to +0.55). Except for subscapular skinfold thickness and length, all neonatal measurements were intermediate between those from Southampton and Pune. The thin-fat phenotype is preserved in Surinam South Asian neonates of the fourth to fifth generation after migration from India. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To determine the incidence and risk factors for neurodevelopmental impairment (NDI) in children with hemolytic disease of the fetus/newborn treated with intrauterine transfusion (IUT). STUDY DESIGN: Neurodevelopmental outcome in children at least 2 years of age was assessed using standardized tests, including the Bayley Scales of Infant Development, the Wechsler Preschool and Primary Scale of Intelligence, and the Wechsler Intelligence Scale for Children, according to the children's age. Primary outcome was the incidence of neurodevelopmental impairment defined as at least one of the following: cerebral palsy, severe developmental delay, bilateral deafness, and/or blindness. RESULTS: A total of 291 children were evaluated at a median age of 8.2 years (range, 2-17 years). Cerebral palsy was detected in 6 (2.1%) children, severe developmental delay in 9 (3.1%) children, and bilateral deafness in 3 (1.0%) children. The overall incidence of neurodevelopmental impairment was 4.8% (14/291). In a multivariate regression analysis including only preoperative risk factors, severe hydrops was independently associated with neurodevelopmental impairment (odds ratio, 11.2; 95% confidence interval, 1.7-92.7). CONCLUSION: Incidence of neurodevelopmental impairment in children treated with intrauterine transfusion for fetal alloimmune anemia is low (4.8%). Prevention of fetal hydrops, the strongest preoperative predictor for impaired neurodevelopment, by timely detection, referral and treatment may improve long-term outcome. [ABSTRACT FROM AUTHOR]

Background: Once a patient has produced a red blood cell (RBC) antibody, there is an increased risk of additional antibody formation after subsequent RBC exposure. Recently, we observed that HLA-DRB1*15 was overrepresented in 379 multiple RBC antibody responders compared to controls or 562 patients with a single RBC antibody (odds ratio [OR], 1.7; 95% confidence interval [CI], 1.3-2.3). In this study we evaluated whether the HLA-DRB1*15 represents a responder phenotype against HLA and/or RBC antigens., Study Design and Methods: HLA-DRB1*15 frequencies in single and multiple antibody responders were compared between three groups of individuals: 1) those with HLA antibodies, 2) those with RBC antibodies, and 3) those with both RBC and HLA antibodies., Results: A total of 3959 immunized patients (female-to-male ratio, 2.3) had been HLA-DRB1 typed. Among the 3275 individuals with HLA antibodies, the frequency of the DRB1*15 phenotype differed significantly from 19.7% in patients with a panel reactivity (PRA) of not more than 20% to 26.9% in patients with PRA of more than 80% (OR, 1.5; 95% CI, 1.2-1.9). This association between DRB1*15 and multiresponsiveness was mainly due to pregnancy-induced HLA immunization. In the 257 individuals with RBC and HLA antibodies, the frequency of DRB1*15 was 4.2 times (95% CI, 1.1-16) higher in those with multiple RBC antibodies and HLA-PRA of more than 50% compared to only single RBC responders with PRA of less than 20%., Conclusion: The HLA-DRB1*15 phenotype is associated with broad RBC and HLA immunization., (© 2016 AABB.)

Background: RhD negativity is distributed unevenly among different ethnicities. In this study we explored the frequencies of RhD negativity in pregnant women in multiethnic Suriname, along with screening results for red blood cell (RBC) antibodies in these women and their offspring. Results may help identify women at risk for hemolytic disease of the fetus and newborn., Study Design and Methods: A retrospective study was performed in pregnant women who delivered at three major hospitals in Suriname between January 1, 2013, and December 31, 2014., Results: The overall prevalences of RhD negativity among 8686 women was 4.3%. The percentages of RhD negativity in Maroons, Creoles, and Hindustani women were 7.2, 5.4, and 3.7%, respectively. Chinese and Javanese women had very low prevalences of 0.8 and 0.5%, respectively, and Amerindians showed no RhD negativity. Antibody screening was positive in six D- (five Maroons and one Creole) women and weakly positive in three women (two Creoles and one Maroon), making overall antibody prevalence 4.4%. In 15 (5.5%; 10 Maroons, four Creoles, one Chinese) newborns from D- mothers antibody screening was positive., Conclusion: In the multiethnic Surinamese population RhD negativity among pregnant women varied between 0.0 and 7.2% between ethnic groups. RBC antibodies were detected during pregnancy and in newborns in 4.4 and 5.5%, respectively., (© 2015 AABB.)

Several studies have shown that South Asian neonates have a characteristic thin-fat insulin-resistant phenotype. The aim of our study was to determine whether this phenotype is also present in South Asians who have migrated to a Western country (the Netherlands). South Asian and white Dutch pregnant women were included in our study. After delivery, cord blood was collected and neonatal anthropometry was measured within 72 h. Compared with white Dutch mothers, South Asian mothers were younger (28.5 v. 32.2 years, P<0.001) and had a higher prepregnancy body mass index (25.1 v. 23.0, P=0.001). Gestational age at delivery was on average 4 days shorter in South Asians (274.9 v. 278.8, P=0.001). To compare the two groups of neonates, we calculated sex- and gestation-specific s.d. scores using the values for mean and s.d. obtained from the white Dutch subjects as a reference. All measurements were smaller in South Asian neonates, except for those of the skinfolds. The largest difference was found in abdominal circumference (s.d. score 1.39, 95% CI -1.76 to -1.01). Triceps and subscapular skinfolds were similar in both groups (triceps s.d. score -0.34, 95% CI -0.88 to +0.20 and subscapular s.d. score -0.03, 95% CI -0.31 to +0.25). South Asian neonates had higher cord plasma levels of triglycerides (0.40 v. 0.36, P=0.614), glucose (5.4 v. 4.8, P=0.079) and insulin (6.3 v. 4.0, P=0.051). However, these differences were not statistically significant. After adjustment for birth weight, the difference in insulin became statistically significant (P=0.001). We therefore conclude that the thin-fat insulin-resistant phenotype is also present in South Asian neonates in the Netherlands.

Objective: To characterise pregnancies where the fetus or neonate was diagnosed with fetal and neonatal alloimmune thrombocytopenia (FNAIT) and suffered from intracranial haemorrhage (ICH), with special focus on time of bleeding onset., Design: Observational cohort study of all recorded cases of ICH caused by FNAIT from the international No IntraCranial Haemorrhage (NOICH) registry during the period 2001-2010., Setting: 13 tertiary referral centres from nine countries across the world., Participants: 37 mothers and 43 children of FNAIT pregnancies complicated by fetal or neonatal ICH identified from the NOICH registry was included if FNAIT diagnosis and ICH was confirmed., Primary and Secondary Outcome Measures: Gestational age at onset of ICH, type of ICH and clinical outcome of ICH were the primary outcome measures. General maternal and neonatal characteristics of pregnancies complicated by fetal/neonatal ICH were secondary outcome measures., Results: From a total of 592 FNAIT cases in the registry, 43 confirmed cases of ICH due to FNAIT were included in the study. The majority of bleedings (23/43, 54%) occurred before 28 gestational weeks and often affected the first born child (27/43, 63%). One-third (35%) of the children died within 4 days after delivery. 23 (53%) children survived with severe neurological disabilities and only 5 (12%) were alive and well at time of discharge. Antenatal treatment was not given in most (91%) cases of fetal/neonatal ICH., Conclusions: ICH caused by FNAIT often occurs during second trimester and the clinical outcome is poor. In order to prevent ICH caused by FNAIT, at-risk pregnancies must be identified and prevention and/or interventions should start early in the second trimester.

Objective: To test the hypothesis that the increased risk of type 2 diabetes mellitus and coronary artery disease in South Asian subjects could be caused by the presence of endothelial dysfunction in early life. We studied markers of endothelial dysfunction in umbilical cord blood of South Asian neonates and compared these with that of Caucasian control subjects., Study Design: From South Asian (n = 57) and Caucasian (n = 21) neonates, cord blood was collected and levels of glucose, insulin, lipids, and markers of endothelial dysfunction (E-selectin, intercellular adhesion molecule 1, vascular cell adhesion molecule 1) and inflammation (C-reactive protein) were measured., Results: Plasma E-selectin levels were significantly higher in South Asian neonates (46.7 versus 33.5 ng/mL, P < .001), and levels of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 did not differ. Furthermore, South Asian neonates had hyperinsulinemia (P = .043), dyslipidemia (with significantly higher triglyceride and lower high-density lipoprotein cholesterol levels), and higher C-reactive protein levels (75.7 versus 43.8 ng/mL, P = .009)., Conclusions: South Asian newborns are characterized by elevated E-selectin levels in line with the hypothesis that endothelial dysfunction is present early in life. In addition, hyperinsulinemia, dyslipidemia, and inflammation are present. Because many pathogenic variables for coronary artery disease and type 2 diabetes are already present at birth in South Asian patients, the question arises whether rigorous childhood lifestyle intervention could be beneficial., (Copyright © 2012 Mosby, Inc. All rights reserved.)

Background: The Leiden University Medical Center (LUMC) is the Dutch national referral centre for pregnancies complicated by haemolytic disease of the fetus and newborn (HDFN) caused by maternal alloimmunization. Yearly, 20-25 affected fetuses with severe anaemia are transfused with intra-uterine blood transfusions (IUT). Mothers of whom their fetus has undergone IUT for HDFN are considered high responders with regard to red blood cell (RBC) antibody formation. Most study groups report high perinatal survival, resulting in a shift in attention towards short- and long-term outcome in surviving children., Methods/design: We set up a large long-term observational follow-up study (LOTUS study), in cooperation with the Sanquin Blood Supply Foundation and the LUMC departments of Obstetrics, Neonatology and ImmunoHematology & Bloodtransfusion.The first part of this study addresses several putative mechanisms associated with blood group alloimmunization in these mothers. The second part of this study determines the incidence of long-term neurodevelopment impairment (NDI) and associated risk factors in children treated with IUT. All women and their life offspring who have been treated with IUT for HDFN in the LUMC from 1987-2008 are invited to participate and after consent, blood or saliva samples are taken. RBC and HLA antigen profile and antibodies are determined by serologic or molecular techniques. Microchimerism populations are tested by real time polymerase chain reaction (RT PCR).All children are tested for their neurological, cognitive and psychosocial development using standardised tests and questionnaires. The primary outcome is neurodevelopmental impairment (NDI), a composite outcome defined as any of the following: cerebral palsy, cognitive or psychomotor development < 2 standard deviation, bilateral blindness and/or bilateral deafness., Discussion: The LOTUS study includes the largest cohort of IUT patients ever studied and is the first to investigate post-IUT long-term effects in both mother and child. The results may lead to a change in transfusion policy, in particular future avoidance of certain incompatibilities. Additionally the LOTUS study will provide clinicians and parents better insights in the long-term neurodevelopmental outcome in children with HDFN treated with IUTs, and may improve the quality of antenatal counselling and long-term guidance.

Prophylactic anti-D is a very safe and effective therapy for the suppression of anti-D immunization and thus prevention of haemolytic disease of the foetus and newborn. However, migration from countries with low health standards and substantial cuts in public health expenses have increased the incidence of anti-D immunization in many "developed" countries. Therefore, this forum focuses on prenatal monitoring standards and treatment strategies in pregnancies with anti-D alloimmunization. The following questions were addressed, and a response was obtained from 12 centres, mainly from Europe.

Objective: The purpose of this study was to determine the long-term neurodevelopmental outcome in fetuses with severe tachy- or bradyarrhythmia., Study Design: This was a follow-up study to assess the neurologic, mental, and psychomotor development in cases with fetal cardiac arrhythmia., Results: A total of 44 fetuses were diagnosed with fetal tachy- or bradyarrhythmia: 28 fetuses had supraventricular tachycardia (SVT); 7 fetuses had atrial flutter (AF), and 9 fetuses had atrioventricular block (AVB). The mortality rate was low (6%; 2/35 fetuses) in the SVT and AF groups and high in the AVB group (78%; 7/9 fetuses). Six patients were lost to follow-up evaluation (14%). Neurodevelopmental outcome was normal in all survivors in the SVT and AF groups, except for 1 patient who experienced plexus brachialis injury because of shoulder dystocia. Two of the 3 survivors in the AVB group had severe developmental delay., Conclusion: The mortality rate and neurodevelopmental impairment in infants with SVT and AF are low, but the mortality rate in infants with AVB is elevated.

Up to 36% of pregnant women are colonized with Group B Streptococcus (GBS). Preterm delivery in colonized mothers is a risk factor for early onset neonatal GBS disease, but whether maternal GBS genital colonization is related to preterm delivery is unclear. The objective of this review was to determine the relationship between maternal colonization with GBS and preterm delivery. Pubmed searches and reference lists of all selected publications were used to find studies reporting on the relationship between maternal GBS colonization and preterm delivery. Study characteristics were abstracted, and validity scores were performed. To assess the relationship between GBS colonization and pregnancy outcome, four-fold prognostic tables were constructed for each study. Out of more than 60 full-text articles, 16 follow-up studies and four case control studies were included in this review. Follow-up studies were divided into 'cohort studies,' in which cultures were taken early in pregnancy and which reported on pregnancy outcome, and 'cross-sectional studies', in which cultures were collected during delivery. Studies differed widely in methods, validity score, and GBS prevalence. The combined estimate from a random effect meta-analysis of the 11 cohort studies was 1.06 (95% confidence intervals (CI) 0.95-1.19) and for the five cross-sectional studies 1.75 (95% CI 1.43-2.14). For the case control studies, the pooled odds ratio was 1.59 (95% CI 1.03-2.44). This systematic review did not show an association between maternal GBS colonization during pregnancy and preterm delivery. However, in case of preterm delivery, there is an increased risk of subsequent maternal GBS colonization.

Protective mechanisms are likely to be present at the fetomaternal interface because fetus-specific alloreactive T cells present in the decidua do not harm the fetus. We tested the immunosuppressive capacity of maternal and fetal multipotent stromal cells (MSC). Single cell suspensions were made from second-trimester amnion, amniotic fluid, and decidua. Culture-expanded cells were identified as MSC based on phenotype and multilineage potential. Coculture of MSC in a primary mixed lymphocyte culture of unrelated responder-stimulator combinations resulted in a dose-dependent inhibition of proliferation. Fetal MSC demonstrated a significantly higher inhibition compared with maternal MSC. This stronger inhibition by fetal MSC was even more prominent in a secondary mixed lymphocyte reaction (MLR) with primed alloreactive T cells. Analysis of cytokine production revealed that fetal MSC produced significantly more interleukin (IL)-10 and vascular endothelial growth factor than maternal MSC. Cell-cell contact is needed for part of the inhibitory effects of MSC. In addition, soluble factors play a role because blocking experiments with anti-IL-10 revealed that the inhibition of the MLR response by fetal MSC is mainly mediated by IL-10. For maternal MSC, other soluble factors seem to be involved. Fetal MSC derived from the fetomaternal interface have a stronger inhibitory effect on naive and antigen-experienced T cells compared with maternal MSC, which is probably related to their higher IL-10 production.

Objectives: To compare fetal cardiac output (CO) in donor and recipient twins of twin-twin transfusion syndrome (TTTS) pregnancies after laser therapy with that of monochorionic twins without TTTS and normal singletons., Methods: In a longitudinal, prospective study, we sonographically assessed fetal CO in donors (n = 10) and recipients (n = 10) with TTTS after fetoscopic laser therapy, in monochorionic twins without TTTS (n = 20) and in normal singleton pregnancies (n = 20). The fetal CO of TTTS twins was determined 1 day and 1 week after laser treatment, and from then on every 2-4 weeks until birth. Twins without TTTS were examined biweekly until birth. Singletons were examined twice, with an 8-week interval, at different gestational ages between 17 and 35 weeks., Results: Absolute CO increased exponentially with advancing gestational age (P < 0.0001), and was significantly related to fetal weight in all groups (P < 0.0001). The median CO/kg in donors after laser therapy, recipients after laser therapy and non-TTTS monochorionic twins was significantly higher than that in singletons (all P < 0.001). Median CO/kg in donors after laser therapy, recipients after laser therapy, and non-TTTS monochorionic twins was not significantly different between groups., Conclusions: Monochorionic twins with TTTS have higher CO after laser therapy than normal singletons., ((c) 2008 ISUOG. Published by John Wiley & Sons, Ltd.)

Objective: The objective of the study was to evaluate the incidence of fetal thrombocytopenia and association with hydrops in Rhesus D alloimmunization., Study Design: The study was a retrospective chart review of 914 intrauterine transfusions in 314 pregnancies performed between 1988 and 2005 in a single institution. The incidence of thrombocytopenia and severity of hydrops at cordocentesis were assessed and correlated with perinatal mortality., Results: Thrombocytopenia (less than 150 x 10(9)/L) was found in 241 of 914 (26%) and severe thrombocytopenia (less than 50 x 10(9)/L) in 25 of 914 (3%) cordocentesis. Twenty-three percent of severely hydropic fetuses had severe thrombocytopenia, compared with 3% and 1% of mildly hydropic and nonhydropic fetuses, respectively. Thrombocytopenia was an independent risk factor for perinatal mortality. Mortality in fetuses that were severely thrombocytopenic and severely hydropic was 67%., Conclusion: Thrombocytopenia is common in hydropic anemic fetuses. Severe thrombocytopenia is associated with a poor prognosis, irrespective of the presence of hydrops. The option of platelet transfusion in severely hydropic anemic fetuses needs further study.

Background and Objectives: Kell haemolytic disease in pregnancies has been suggested to be associated with decreased fetal platelet counts. The aim of this study was to evaluate the incidence and clinical significance of fetal thrombocytopenia in pregnancies complicated by Kell alloimmunization., Materials and Methods: In this retrospective cohort study, fetal platelet counts were performed in 42 pregnancies with severe Kell alloimmunization prior to the first intrauterine blood transfusion. Platelet counts from 318 first intrauterine transfusions in RhD alloimmunized pregnancies were used as controls., Results: Fetal thrombocytopenia (platelet count < 150 x 10(9)/l) was found in 4/42 (10%) in the Kell group and in 84/318 (26%) in the RhD group. None of the fetuses in the Kell alloimmunized pregnancies, including 15 with severe hydrops, had a clinically significant thrombocytopenia defined as a platelet count < 50 x 10(9)/l. In the RhD alloimmunized pregnancies, 2/230 (1%) of the non-hydropic fetuses and 7/30 (23%) of the severely hydropic fetuses had a clinically significant thrombocytopenia., Conclusion: In contrast to fetuses with severe anaemia and hydrops due to RhD alloimmunization, fetuses with severe anaemia due to Kell alloimmunization are generally not at risk for substantial thrombocytopenia.

The two regions of the maternal decidua, decidua basalis and decidua parietalis, differ in the extent of trophoblast invasion and consequently in cytokines and other biological mediators, extracellular matrix and cellular components. Our aim was to compare the phenotypic features of macrophages from the two decidual regions across a broad gestational age range. We isolated macrophages by enzymatic digestion from healthy decidua samples obtained after elective abortions, at 9-18-week and at 19-23-weeks, or after term deliveries (caesarean sections at term and spontaneous term vaginal deliveries). Macrophages were analysed by flow cytometry applying the same instrument settings to all the samples to allow semi-quantitative comparison of the expression of a particular marker between different samples. We found higher expressions of CD80, CD86 and HLA-DR, suggestive of a more activated phenotype of decidual macrophages, at early/mid pregnancy than at term. Marginal differences were found between term decidual macrophages obtained after spontaneous vaginal deliveries or caesarean sections which imply that the parturient process is not associated with decidual macrophage activation. The expressions of CD105, DC-SIGN and MMR were the strongest in decidua basalis of mid pregnancy and indicate the importance of decidual macrophages in tissue homeostasis at the uteroplacental interface.

Background: In 1998 a national program for first-trimester screening for red cell (RBC) antibodies in all pregnant women was implemented. The aim of our study was to assess the impact on perinatal mortality caused by Kell alloimmunization, Study Design and Methods: Prospectively collected data on all pregnant women referred to our center from 1988 until 2005 for intrauterine transfusion (IUT) for fetal anemia due to Kell alloantibodies were analyzed. The cohort was divided into two groups, those treated before 1998 and those treated after 1998. The primary outcome was fetal and neonatal survival. Secondary outcome variables were gestational age, fetal hemoglobin (Hb) levels at first IUT, severity of hydrops, and total number of IUTs per pregnancy. Causes for mortality were analyzed in detail., Results: A total of 43 pregnancies were included, 18 before introduction of screening and 25 thereafter. Perinatal survival increased from 61 percent in the first period to 100 percent after introduction of screening. After 1998, fetal hydrops was generally less severe at first IUT, while gestational age and fetal Hb levels at first IUT were similar., Conclusion: Implementation of routine screening for Kell antibodies in pregnancy was associated with an increased referral rate for suspected fetal anemia, more timely referrals, and a higher perinatal survival rate after intrauterine treatment.

Objectives: To evaluate the influence of fetoscopic laser therapy on fetal cardiac size in monochorionic twins complicated by twin-twin transfusion syndrome (TTTS)., Methods: In a longitudinal, prospective study, we assessed fetal cardiac size sonographically in monochorionic diamniotic twins with TTTS treated by laser therapy and in monochorionic twins without TTTS. The fetal cardiothoracic ratio (cardiac circumference divided by thoracic circumference) of TTTS twins was determined within 24 h before, 12-24 h after and 1 week after laser treatment, and from then on every 2-4 weeks until birth. TTTS twins were classified into Quintero Stages 1-2 (n = 18) and Stages 3-4 (n = 16), and measurements were compared with biweekly measurements of non-TTTS monochorionic twins matched for gestational age (n = 38). Cardiomegaly was defined as a cardiothoracic ratio above the 97.5th percentile., Results: Before laser treatment, cardiomegaly was observed in 44% (8/18) and 50% (8/16) of recipients in Quintero Stages 1-2 and Stages 3-4, respectively. Cardiomegaly occurred in none of the donors before treatment. After laser treatment, cardiomegaly was observed in 76% (13/17) and 50% (7/14) of recipients in Stages 1-2 and Stages 3-4, respectively, and in 17% (3/18) and 13% (2/15) of donors in Stages 1-2 and Stages 3-4, respectively. Cardiomegaly was present in 18% (7/38) and 8% (2/25) of non-TTTS monochorionic twins and singletons. After laser therapy, the cardiothoracic ratio of recipients in Stages 1-2 and Stages 3-4 was not significantly changed (P = 0.34 and P = 0.67, respectively). The cardiothoracic ratio of donors in Stages 1-2 and Stages 3-4 was increased compared with that before laser therapy (P = 0.0002 and P = 0.005, respectively). Cardiothoracic ratios of non-TTTS monochorionic twins were not significantly different from our reference range in singletons throughout gestation, and were smaller than those in both recipients and donors after laser therapy., Conclusions: TTTS recipients show cardiomegaly before as well as after fetoscopic laser therapy for TTTS. Donors develop cardiomegaly only after laser treatment. Our findings emphasize the significant effect of TTTS and fetoscopic laser therapy on the fetal heart of both recipient and donor twins., (Copyright (c) 2008 ISUOG. Published by John Wiley & Sons, Ltd.)

Objectives: In monoamniotic twin pregnancies discordant for fetal anomaly, parents may opt for selective feticide. However, the normal co-twin remains at risk of sudden demise from cord entanglement. We report on three cases of successful selective feticide by cord occlusion combined with cord transection., Methods: We describe technical details and outcome of three monoamniotic twin pregnancies discordant for fetal anomaly (two cases of anencephaly and one case of congenital heart block) in which cord occlusion was followed by transection of the cord using contact laser., Results: The fetoscopic cord occlusion and transection using laser was successfully performed at 15, 16 and 19 weeks gestation, respectively. In one case, amniotic fluid leakage occurred after fetoscopy. The surviving co-twins were born at 36, 38 and 36 weeks gestation, respectively; two of the three were born vaginally and they were all healthy., Conclusion: In monoamniotic twins, selective feticide using laser occlusion and transection of the umbilical cord is technically feasible and can lead to near-term vaginal birth of healthy co-twins., (Copyright 2007 S. Karger AG, Basel.)

Objective: To study the short-term morbidity in Rhesus hemolytic disease of infants treated either with or without intrauterine transfusions (IUT)., Study Design: All term and near term infants (gestational age > or = 36 weeks) with neonatal Rhesus hemolytic disease admitted to our center between January 2000-March 2005 were retrospectively included in the study. We recorded the duration of phototherapy, the need of exchange transfusions, and the need of top-up red blood cell transfusions until 6 months of age., Results: A total of 89 infants were included, of whom 52 received at least one IUT. Duration of phototherapy in the IUT and no-IUT group was 3.8 and 5.1 days, respectively (P = .01). The percentage of infants requiring an exchange transfusion in the IUT group was 71% compared to 65% in the no-IUT group (P = .64). The percentage of infants requiring a top-up transfusion in the IUT and no-IUT group was 77% and 26.5%, respectively (P < .01)., Conclusion: Infants with Rhesus hemolytic disease treated with IUT required less days of phototherapy and more top-up red blood cell transfusions than neonates without IUT. However, the need for exchange transfusion was similar in both groups.

We evaluated the effects of platelet transfusions and intravenous immunoglobulin (IVIG) in neonates with fetal and neonatal alloimmune thrombocytopenia (FNAIT) with and without antenatal treatment with IVIG. Records of neonates with FNAIT admitted between January 2000 and November 2005 were reviewed. The patients were divided into group I, treated antenatally with IVIG for known FNAIT, and group II, postnatally diagnosed with FNAIT. The primary outcome was the time interval to reach a platelet level above 100 x 10(9)/L in relation to the type of treatment. Nineteen neonates with FNAIT were identified, 13 in group I and 6 in group II. In group I, four children were born with a platelet count above 100 x 10(9)/L and never needed treatment, and four received a single matched platelet transfusion at birth with a maintained response. Five neonates received IVIG and one matched transfusion, with all but one rapidly responding. In antenatally treated cases, postnatal IVIG had no apparent effect on the platelet count. In group II, two neonates died on day 1 with severe intracranial hemorrhage. Two of the four other patients responded to a number of unmatched platelet transfusions, with one neonate rapidly responding after one matched transfusion, while another needed nine matched transfusions before a persistent adequate platelet count was reached after 9 weeks. Postnatal IVIG had no apparent effect on the platelet count in any of our cases. In neonates with FNAIT treated antenatally with IVIG, neonatal management using a single matched platelet transfusion was adequate in all cases. In neonatally diagnosed cases not treated before birth, multiple matched platelet transfusions may be required. We found no evidence to support the use of IVIG in neonates with FNAIT.

Objective: To compare fetoscopic laser surgery with amniodrainage in the treatment of twin-to-twin transfusion syndrome (TTTS) diagnosed after 26 weeks of gestation., Design: A retrospective cohort study., Setting: Leiden University Medical Centre, a tertiary referral hospital for fetal therapy., Population: Between January 1991 and February 2006, 21 TTTS cases were diagnosed and treated after 26 weeks of gestation., Methods: Treatment of TTTS consisted of either amniodrainage or fetoscopic laser coagulation of vascular anastomoses., Primary Outcome: adverse outcome (intrauterine or neonatal death, major neonatal morbidity and/or severe cerebral injury). Secondary outcome: gestational age at birth., Results: Eleven TTTS cases were treated with amniodrainage and ten with laser surgery. Median gestational age at birth in the amniodrainage group and in the laser surgery group was 29 and 31 weeks, respectively (P = 0.17) All infants were born alive. Major neonatal morbidity occurred more often in the amniodrainage group than in the laser surgery group, 27% (6/22) and 0% (0/20), respectively (P = 0.02). Severe cerebral injury in the amniodrainage group and in the laser surgery group occurred in 23% (5/22) and 15% (3/20) of infants, respectively (P = 0.70). Neonatal mortality in the amniodrainage group and in the laser surgery group was 14% (3/22) and 0% (0/20), respectively (P = 0.23). Overall adverse outcome was 36% (8/22) in the amniodrainage group and 15% (3/20) in the laser surgery group (P = 0.17)., Conclusion: In TTTS diagnosed after 26 weeks of gestation, amniodrainage and laser surgery both result in 100% survival. However, infants born after laser surgery have less major neonatal morbidity.

Objective: To describe the outcome of pregnancies with fetal and neonatal alloimmune thrombocytopenia (FNAIT) in relation to the invasiveness of the management protocol., Design: Retrospective analysis of prospectively collected data from a national cohort., Setting: Leiden University Medical Centre, the national centre for management of severe red cell and platelet alloimmunisation in pregnancy., Population: Ninety-eight pregnancies in 85 women with FNAIT having a previous child with thrombocytopenia with (n= 16) or without (n= 82) an intracranial haemorrhage (ICH)., Methods: Our management protocol evolved over time from (1) serial fetal blood samplings (FBS) and platelet transfusion (n= 13) via (2) combined FBS with maternal intravenous immunoglobulins (n= 33) to (3) completely noninvasive treatment with immunoglobulins only (n= 52 pregnancies, resulting in 53 neonates). Perinatal outcome was assessed according to the three types of management., Main Outcome Measures: Occurrence of ICH, perinatal survival, gestational age at birth and complications of FBS., Results: All but one of 98 pregnancies ended in a live birth; none of the neonates had an ICH. The median gestational age at birth was 37 weeks (range 32-40). In groups 1 and 2, three emergency caesarean sections were performed after complicated FBS, resulting in two healthy babies and one neonatal death., Conclusion: Noninvasive antenatal management of pregnancies complicated by FNAIT appears to be both effective and safe.

Objective: Intrauterine transfusion (IUT) is a life-saving therapy for the severely anemic fetus with hemolytic disease. However, maternal additional antibody formation is a complication of the procedure. In this study, we determined antibody formation after introduction of preventive Rh-D, -C, -c, -E, and -e and K matching of IUT donors., Study Design: This was a retrospective follow-up study., Results: During an 11-year period, 686 Rhesus- and K-matched IUTs were performed in 233 pregnancies and in 95% (652/686) posttransfusion antibody testing was performed after a median interval of 21 days. Twenty-five percent (53/212) of the women formed 64 new antibodies and, compared to our previous study, this incidence was not decreased by the use of Rhesus- and K-matched donors. After delivery, 72% (153/212) of the women had multiple RBC antibodies. Additional antibodies were in 48% (31/64) directed against Rhesus and K antigens, induced by the fetus, or as natural antibodies. In 52% (33/64) the antibodies were directed against non-Rhesus and -K antigens and in 65% (11/17) of eligible cases the IUT donor and not the fetus expressed the corresponding antigen(s)., Conclusion: Despite Rhesus- and K-matching, women treated with IUTs still show strikingly broad red cell alloimmunization. More extensive IUT donor red cell matching, including FY, JK, and S antigens, to reduce the formation of new red cell antibodies should be explored.

Placental vascular anastomoses in monochorionic twins may lead to acute or chronic inter-twin transfusion. We report an uncommon form of chronic inter-twin transfusion, referred to as twin anemia-polycythemia sequence (TAPS), with severe anemia in one twin and polycythemia in the other, without the characteristically associated twin oligo-polyhydramnios sequence (TOPS) seen in the classical twin-to-twin transfusion syndrome (TTTS). The clinical course and placental characteristics of two pairs of monochorionic twins with TAPS born at, respectively, 33 and 34 weeks' gestation were reviewed. Serial fetal ultrasound examinations revealed no signs of TOPS. At birth, both donor twins were severely anemic requiring blood transfusion and both recipients were polycythemic, one requiring partial volume exchange transfusions. Inter-twin difference in reticulocyte counts was extremely high, suggesting a chronic form of inter-twin blood transfusion. Placental injection studies revealed a preponderance of very small (<1mm) arterio-venous anastomoses in one direction. In conclusion, chronic inter-twin transfusion may lead to an uncommon form of inter-twin transfusion, named TAPS, not associated with TOPS, resulting in severe fetal or neonatal hematological complications. We hypothesize that TAPS is mediated through minuscule unidirectional anastomoses. TAPS can be diagnosed antenatally with Doppler studies and postnatally by hemoglobin and reticulocyte measurements.

Objective: In this prospective cohort study, we evaluated the initial results of fetoscopic laser surgery for severe second trimester twin-to-twin transfusion syndrome (TTTS) treated at our centre., Method: A total of 100 consecutive pregnancies with severe second trimester TTTS treated at our centre with selective fetoscopic laser coagulation of vascular anastomoses on the placental surface between August 2000 and November 2004 were included in the study. Perinatal survival was analysed in relation to Quintero stage., Results: Median gestational age was 20 weeks at fetoscopy (range: 16-26) and 33 weeks at delivery (range: 18-40). Perinatal survival rate was 70% (139/200). The treatment resulted in at least one survivor at the age of 4 weeks in 81% of pregnancies. Perinatal survival was significantly higher when treatment was performed in the early Quintero stages (95% in stage 1, 76% in stage 2, 70% in stage 3, 50% in stage 4) (p = 0.02)., Conclusion: Results of fetoscopic laser surgery for TTTS in our centre are similar to those in specialised centres in other countries. Diagnosis and treatment in the early Quintero stages resulted in significantly higher perinatal survival., ((c) 2007 S. Karger AG, Basel.)

Objectives: To assess the value of serial ultrasound examinations together with patient instructions to report the onset of symptoms in achieving timely detection of twin-to-twin transfusion syndrome (TTTS) in a cohort of monochorionic diamniotic twin pregnancies, and to evaluate sonographic TTTS predictors., Methods: Timely detection of TTTS was defined as diagnosis before severe complications of TTTS occurred, such as preterm prelabor rupture of membranes, very preterm delivery (24-32 weeks of pregnancy), fetal hydrops, or intrauterine fetal death. During a 2-year period, a prospective series of 23 monochorionic twin pregnancies was monitored from the first trimester until delivery. At least every 2 weeks we performed ultrasound and Doppler measurements (nuchal translucency thickness, presence of membrane folding, estimated fetal weight, deepest vertical pocket, bladder filling, and Doppler waveforms of the umbilical artery, ductus venosus and umbilical vein). Measurements of TTTS cases were compared with those of non-TTTS cases matched for gestational age. Furthermore, patients were informed about the symptoms caused by TTTS, and instructed to consult us immediately in case of rapidly increasing abdominal size or premature contractions., Results: In all four TTTS cases, the diagnosis was timely. At the time of diagnosis, one case was at Quintero Stage 1, two at Quintero Stage 2, and one at Quintero Stage 3. Two of the TTTS cases became apparent after the patients' feeling of rapidly increasing girth. The identification of TTTS predictors was successful with respect to one parameter: isolated polyhydramnios in one sac, without oligohydramnios in the other, preceded the ultimate diagnosis of TTTS in two of the four TTTS cases. All other ultrasound measurements of TTTS cases, prior to the diagnosis of TTTS, were within the range of measurements of non-TTTS cases., Conclusion: Biweekly ultrasound examinations, with special attention to the amniotic fluid compartments of both fetuses, combined with detailed patient instructions to report the onset of symptoms resulted in timely diagnosis of all TTTS cases and appears to be a safe program for monitoring monochorionic twin pregnancies., (Copyright 2006 ISUOG. Published by John Wiley & Sons, Ltd.)

Background: Pregnancies complicated by Rh alloimmunization have been evaluated with the use of serial invasive amniocentesis to determine bilirubin levels by measuring in the amniotic fluid the change in optical density at a wavelength of 450 nm (DeltaOD450); however, this procedure carries risks. Noninvasive Doppler ultrasonographic measurement of the peak velocity of systolic blood flow in the middle cerebral artery also predicts severe fetal anemia, but this test has not been rigorously evaluated in comparison with amniotic-fluid DeltaOD450., Methods: We performed a prospective, international, multicenter study including women with RhD-, Rhc-, RhE-, or Fy(a)-alloimmunized pregnancies with indirect antiglobulin titers of at least 1:64 and antigen-positive fetuses to assess whether Doppler ultrasonographic measurement of the peak systolic velocity of blood flow in the middle cerebral artery was at least as sensitive and accurate as measurement of amniotic-fluid DeltaOD450 for diagnosing severe fetal anemia. The results of the two tests were compared with the incidence of fetal anemia, as determined by measurement of hemoglobin levels in fetal blood., Results: Of 165 fetuses, 74 had severe anemia. For the detection of severe fetal anemia, Doppler ultrasonography of the middle cerebral artery had a sensitivity of 88 percent (95 percent confidence interval, 78 to 93 percent), a specificity of 82 percent (95 percent confidence interval, 73 to 89 percent), and an accuracy of 85 percent (95 percent confidence interval, 79 to 90 percent). Amniotic-fluid DeltaOD450 had a sensitivity of 76 percent (95 percent confidence interval, 65 to 84 percent), a specificity of 77 percent (95 percent confidence interval, 67 to 84 percent), and an accuracy of 76 percent (95 percent confidence interval, 69 to 82 percent). Doppler ultrasonography was more sensitive, by 12 percentage points (95 percent confidence interval, 0.3 to 24.0), and more accurate, by 9 percentage points (95 percent confidence interval, 1.1 to 15.9), than measurement of amniotic-fluid DeltaOD450., Conclusions: Doppler measurement of the peak velocity of systolic blood flow in the middle cerebral artery can safely replace invasive testing in the management of Rh-alloimmunized pregnancies. (ClinicalTrials.gov number, NCT00295516.)., (Copyright 2006 Massachusetts Medical Society.)

Objectives: To evaluate the safety of vaginal delivery in pregnancies with fetal and neonatal alloimmune thrombocytopenia (FNAIT)., Design: Prospective data collection., Setting: Leiden University Medical Centre, the national centre for management of severe red cell and platelet alloimmunisation., Population: Thirty-two pregnancies with FNAIT, with a sibling with thrombocytopenia but without an intracranial haemorrhage (ICH)., Methods: The mode of delivery, platelet count in cord blood and neonatal outcome were analysed. All women received weekly intravenous immunoglobulin from 32 to 38 weeks of gestation. Head ultrasound scan was performed in all neonates., Main Outcome Measures: Signs of ICH or other bleeding in the neonates., Results: Twenty-three women delivered vaginally. Nine caesarean sections were performed, all for obstetric reasons. Median platelet count at birth was 142 x 10(9)/l (range, 4-252 x 10(9)/l), with severe thrombocytopenia (<50 x10(9)/l) in four neonates, of which three were born vaginally. None of the neonates showed signs of ICH or other bleeding., Conclusions: In pregnancies with FNAIT and a thrombocytopenic sibling without ICH, vaginal delivery was not associated with neonatal intracranial bleeding. These initial results support our noninvasive management of these pregnancies with FNAIT.

During pregnancy several maternal and fetal mechanisms are established to prevent a destructive immune response against the allogeneic fetus. Despite these mechanisms, fetus specific T-cells persist throughout gestation but little is known about the regulation of these T-cells. Recently, CD4(+)CD25(+) regulatory T-cells have been identified in human decidua. Human decidua forms the maternal part of the fetal-maternal interface and is subdivided in two distinct regions: the decidua (d.) basalis and the decidua (d.) parietalis. The aim of this study was to determine the distribution of specific T-cell subsets in d. basalis and d. parietalis in early and term pregnancy, with a special emphasis on the presence of CD4(+)CD25(bright) (regulatory) T-cells and CD8(+)CD28(-) (suppressor) T-cells. In addition, we compared phenotypic characteristics of decidua derived T-cell subsets with maternal peripheral blood (mPBL) T-cells and T-cells from non-pregnant controls. We identified significantly higher percentages of CD4(+)CD25(bright) and CD8(+)CD28(-) T-cells in decidua compared to peripheral blood suggesting an important role for these T-cell subsets locally at the fetal-maternal interface. The major differences in T-cell subset distribution and the presence of additional phenotypic differences between T-cells in d. basalis, d. parietalis and mPBL may reflect specific immunomodulatory functions of these T-cell subsets at these different sites during pregnancy.

Objective: To investigate whether chorionic villus sampling (CVS) results in a proportional increase of alpha-fetoprotein (AFP) and fetal red cells in maternal blood., Methods: Blood samples were collected before and after CVS. The AFP concentration was measured and supervised automated microscopy of Kleihauer-Betke slides was applied to quantify fetal red cells., Results: AFP analysis was performed in 53 paired samples and automated microscopic scanning in 59 paired samples. Median AFP concentrations before and after CVS were 12.0 microg/L (range 6.4-36.4) and 18.7 microg/L (range 8.2-668.9), respectively, indicating a significant increase (p < 0.0001). Median numbers of fetal red cells detected before and after CVS were 0 (range 0-36) and 0 (range 0-31), respectively. No significant increase of fetal cells was observed (p = 0.72). The delta (Delta) fetal red cells and the Delta AFP correlated poorly (rho = -0.22, p = 0.11). The amount of villi correlated moderately with the Delta AFP (rho = 0.32, p = 0.02) and did not correlate with the Delta fetal red cells (rho = -0.11, p = 0.43)., Conclusions: Although the AFP concentration after CVS increased, no increase of fetal red cells was detected. These findings suggest that CVS results in a leakage of proteins due to placental tissue damage, rather than increased trafficking of fetal cells., (2006 John Wiley & Sons, Ltd.)

Objective: To report on a less invasive treatment strategy in alloimmune fetal and neonatal thrombocytopenia (FNAIT) at high risk for either in utero or neonatal intracranial hemorrhage (ICH)., Methods: In 7 pregnancies, with a history of ICH in the older sibling, weekly intravenous immunoglobulin (IVIG) therapy to the mother (1 g/kg) without initial cordocentesis was started at a median gestational age of 16 weeks., Results: In 4 pregnancies cordocentesis was avoided. One predelivery cordocentesis with platelet transfusion was performed in 3 further cases. Although none of the cases had a platelet count of >50 x 10(9)/l at cordocentesis, predelivery or birth, no ICHs were observed. The neonatal periods of the infants were uncomplicated., Conclusion: IVIG treatment alone might be considered in patients with both severe platelet alloimmunization and an increased risk for morbidity and mortality at cordocentesis., ((c) 2006 S. Karger AG, Basel)

Objective: To evaluate the effect of fetal anemia and intrauterine transfusion on ventricular shortening fraction., Methods: The end-diastolic and end-systolic transverse dimensions of the left and right ventricles were obtained using M-mode ultrasonography. The shortening fractions of both ventricles were calculated at three time points: before, immediately after and one day after intrauterine transfusion. The blood volume given at intrauterine transfusion was expressed as a percentage of estimated fetoplacental blood volume., Results: Complete measurements were obtained from 49 transfusions in 23 fetuses. Intrauterine transfusion was performed at a median gestational age of 31 (range, 19-35) weeks. Median hemoglobin concentration before and after intrauterine transfusion was 7.9 (range, 2.7-13.7) g/dL and 14.3 (range, 12.7-16.1) g/dL, respectively. Both left and right ventricular shortening fractions differed significantly between the three time points. Left ventricular shortening fraction decreased immediately after transfusion in 43 (88%) of the 49 procedures. Right ventricular shortening fraction decreased immediately after transfusion in 42 (86%) of the 49 procedures. At the first intrauterine transfusion, there was only a weak correlation between the decrease in shortening fraction of both ventricles and the transfused volume (left: R(2) = 0.15; P = 0.20/right: R(2) = 0.005; P = 0.81)., Conclusion: Transfusion significantly decreases the shortening fraction of both ventricles of the fetal heart. There is, however, little correlation between the decrease in shortening fraction and the volume of red cells given at intrauterine transfusion. Published by John Wiley & Sons, Ltd., (Copyright (c) 2005 ISUOG.)

Objective: To compare the electrolyte composition of pregnancies complicated with twin-twin transfusion syndrome (TTTS) with that of physiologic pregnancies., Materials and Methods: Amniotic fluid samples from 16 pregnancies were studied. Specimens were obtained from recipient sacs in 10 pregnancies undergoing fetoscopy for severe midtrimester TTTS. Additionally, 6 amniotic fluid samples were obtained transcervically from legal second-trimester pregnancy terminations. The concentrations of sodium, potassium, chloride, bicarbonate, calcium, glucose, osmolality, pH, total protein content and albumin were determined in each sample., Results: The mean gestational age at sampling was 20.2 weeks (range 17.2-27.1) in the TTTS group and 18.4 (range 16.0-22.0) in the control group (p = NS). We found significant lower levels of albumin (0.22 +/- 0.04 vs. 0.39 +/- 0.11, p = 0.01) and total protein (0.19 +/- 0.08 vs. 0.51 +/- 0.17, p < 0.001) and higher levels of bicarbonate (16.90 +/- 1.45 vs. 14.50 +/- 2.17, p = 0.02) in amniotic fluid samples taken from recipient sacs of TTTS pregnancies., Conclusion: Amniotic fluid from the receptor in severe midtrimester TTTS differs significantly from control amniotic fluid samples in bicarbonate concentration, total protein content and albumin concentration. These findings may help to understand the pathophysiology of TTTS and to optimise therapeutic modalities., (Copyright (c) 2005 S. Karger AG, Basel.)

Background and Objectives: Characterization of the different sites of fetal hematopoiesis during the second trimester of pregnancy can provide important information for the timing of in utero stem cell transplantation (SCT), as an experimental treatment for congenital hematologic disorders., Design and Methods: We analyzed the distribution of the different hematopoietic precursor cells in fetal blood, liver, bone marrow (BM), spleen and thymus from 66 fetuses between the ages of 13 to 23 weeks of gestation by flow cytometry and culture of hematopoietic progenitor cells (HPC) in semi-solid media., Results: During the second trimester the percentages of CD34+ cells did not change and were 4.0% (1.0-12.0%) (median [min.-max.]) in blood, 16.5% (3.0-32.0%) in BM, 6.0% (2.0-16.0%) in liver, 5.0% (2.0-14.0%) in spleen, and 1.1% (0.9-3.0%) in the thymus. Each tissue contained all subsets of CD34+ cells at various levels. Within the CD34+ population, in BM the main sub-population was CD34+CD19+ (38% (11-67%)), in thymus CD34+ CD7+ (83% [45-98%]), and in blood and liver CD34+ CD33+ (57% (30-80%) and 48% (20-82%), respectively). In all tissues approximately 1 % of nucleated cells were non-committed CD34+ CD38- cells. The frequencies of both committed CD34+ cells and non-committed CD34+ CD38- cells were constant from 13 to 23 weeks in fetal blood, BM, liver and spleen. The frequencies of cultured HPC were high in fetal liver, low in fetal BM, and increasing in fetal blood., Interpretation and Conclusions: During the second trimester of gestation, all CD34+ subsets were present in each hematopoietic compartment at different levels. An exchange of stem cells between organs is likely, but no major shift of the hematopoietic stem cell compartment from the liver to other hematopoietic organs was found during the mid-trimester. No arguments for a specific time window for performing in utero SCT were found, but if engraftment of donor stem cells in the human fetus is influenced by competition of endogenous stem cells or fetal immune competence, in utero SCT should be performed as early as possible during fetal development.

Objective: The purpose of this study was to establish the true procedure-related complication rate of intrauterine transfusion therapy., Study Design: A cohort study of 254 fetuses treated with 740 intrauterine blood transfusions for red-cell alloimmunization in a single center in the years 1988 to 2001. Our database was searched for perinatal deaths, emergency deliveries, infections, and preterm rupture of membranes associated with intrauterine blood transfusion. Complications were categorized by two independent obstetricians as procedure-related (PR) or not procedure-related (NPR). Logistic regression analysis was used to identify risk factors for complications., Results: Overall survival was 225/254 (89%). Fetal death occurred in 19 cases (7 PR) and neonatal death in 10 cases (5 PR). There were two cases of intrauterine infection with Escherichia coli (both PR) and two other cases of preterm premature rupture of membranes (1 PR) within a week of a procedure. Emergency delivery after a transfusion was performed in 18 pregnancies (15 PR). The total PR complication rate was 3.1%, resulting in an overall PR loss rate of 1.6% per procedure. Arterial puncture, transamniotic cord puncture, refraining from fetal paralysis, and advancing gestational age were associated with the occurrence of PR complications., Conclusion: Our study shows that intrauterine transfusion is a safe procedure, with a relatively low PR perinatal loss rate. Arterial puncture and transamniotic cord needling carry a high risk for serious complications, whereas fetal paralysis improves the safety of the procedure. This information on risks of intrauterine transfusion therapy may help to further improve the safety of intrauterine transfusions. Data on complication rates of intrauterine transfusions are essential in counseling patients.

Objective: To study the effects of oral carbohydrate ingestion on clinical outcome and on maternal and fetal metabolism., Design: Prospective, double-blind, randomised study., Setting: Leyenburg Hospital, The Hague, The Netherlands., Population: Two hundred and two nulliparous women., Methods: In labour, at 8 to 10 cm of cervical dilatation, the women were asked to drink a solution containing either 25 g carbohydrates or placebo. In a subgroup of 28 women, metabolic parameters were measured., Main Outcome Measures: Number of instrumental deliveries, fetal and maternal glucose, free fatty acids, lactate, pH, Pco2, base excess/deficit and beta-hydroxybutyrate., Results: Drinking a carbohydrate-enriched solution just before starting the second stage of labour did not reduce instrumental delivery rate (RR 1.1, 95% CI 0.9-1.3). Caesarean section rate was lower in the carbohydrate group, but the difference did not reach statistical significance (1% vs 7%, RR 0.2, 95% CI 0.02-1.2). In the carbohydrate group, maternal free fatty acids decreased and the lactate increased. In the umbilical cord there was a positive venous-arterial lactate difference in the carbohydrate group and a negative one in the placebo group, but the differences in pH and base deficit were comparable., Conclusion: Intake of carbohydrates just before the second stage does not reduce instrumental delivery rate. The venous-arterial difference in the umbilical cord suggested lactate transport to the fetal circulation but did not result in fetal acidaemia.

Objective: To evaluate the long-term follow-up of surviving offspring after antenatal treatment for fetal or neonatal alloimmune thrombocytopenia (FNAIT)., Patients: Fifty children at risk of FNAIT were antenatally treated with maternal intravenous immunoglobulins (IVIG) (n=11), IVIG with intrauterine platelet transfusions (IUPT) (n=26) or IUPT alone (n=9). In four cases (n=4), only fetal blood sampling (FBS) was performed. One child died in the neonatal period and one was lost to follow up., Methods: The remaining 48 children, aged 1.3-11.6 years (median 5.1 years), were given both general and neurological examinations and assessed on their development and susceptibility for infections or atopic constitution. In addition, immunoglobulin levels were measured in 17 infants, aged 5 years and older., Results: Intracranial hemorrhage (ICH) was not observed. The general health and neurodevelopmental outcome in the children was comparable to a normal Dutch population. Children not exposed to maternal IVIG treatment had significantly more infections and hearing problems than children exposed to IVIG treatment or the normal population. Immunoglobulin G, A and M levels were within the normal range, independent of treatment and severity of FNAIT. A high IgE level was more frequently seen in children exposed to IVIG, but did not result in clinical consequences such as allergy or atopy., Conclusions: Antenatal treatment of children for FNAIT did not affect general health or neurodevelopmental outcome. In particular, exposure to IVIG in utero showed no adverse effect on the clinical outcome of these children.

Objective: The purpose of this study was to evaluate whether maternally administered intravenous immunoglobulins (IVIG) and intrauterine platelet transfusions (IUPT) for fetal/neonatal alloimmune thrombocytopenia (FNAIT) affect the development of the fetal immune system., Study Design: The lymphocyte subset distribution of mononuclear cells of cord blood of 20 FNAIT newborns was analyzed by flow cytometry and compared with a control group of healthy newborns and a reference group treated with intrauterine erythrocyte transfusions (IUET) for hemolytic disease., Results: The percentage of monocytes, natural killer (NK) cells, ratios of mature and immature T cells and B cells, and resting or activated cells were not significantly different compared to the control group. In addition, the B-cell and T-cell populations showed a normal in vitro antibody production and T-cell proliferation when compared with the control group., Conclusion: Antenatal treatment for FNAIT with maternal IVIG with or without IUPT is not associated with lymphocyte activation or premature maturation of the neonatal immune system.

Objective: The purpose of this study was to evaluate the quantification of fetomaternal hemorrhage by the manual and automated microscopic analysis of Kleihauer-Betke stained slides and by flow cytometry., Study Design: Blood smears were stained and evaluated manually according to the Kleihauer-Betke test. The same slides were used for automated microscopy. In addition, blood flow cytometry was performed by anti-hemaglobin F immunostaining., Results: Fetomaternal hemorrhage >0.1% was detected in 4 patients by manual and automated Kleihauer-Betke test and by blood flow cytometry. Fetomaternal hemorrhage was absent according to all 3 methods in 13 patients; fetomaternal hemorrhage<0.1% was detected in 27 patients by either manual or automated Kleihauer-Betke test or both. Moderate agreement was observed between the manual and automated Kleihauer-Betke test (weighted kappa, 0.56; 95% CI, 0.33-0.78). Agreement between the manual Kleihauer-Betke test and blood flow cytometry was fair (weighted kappa, 0.40; 95% CI, 0.15-0.66)., Conclusion: Automated microscopic detection of fetal blood cells in clinical samples provides accurate quantification that is comparable to the manual Kleihauer-Betke test in both small and large fetomaternal hemorrhage. Blood flow cytometry is capable only of quantifying fetomaternal hemorrhage of >0.1%.

Objective: To assess pregnancy outcome after intrauterine transfusion (IUT) for fetal anemia due to red-cell alloimmunization in the Netherlands over 11 years, in order to improve care and counseling., Methods: A retrospective cohort study was conducted from January 1, 1988, to January 1, 1999. Data were collected prospectively on all red-cell alloimmunized pregnancies requiring intrauterine blood transfusions. Primary outcome variables were fetal and neonatal survival in relation to the type of antibody, gestational age and presence or absence of hydrops. In addition, we studied short-term neonatal morbidity and procedure-related complications., Results: A total of 210 fetuses from 208 pregnancies received 593 transfusions. Overall survival rate was 86%. Survival of hydropic fetuses (78%) was significantly different from those without hydrops (92%). Low survival rates were especially found in hydropic fetuses with the first transfusion at gestational ages of 20 weeks or less (55%) or between 28 and 32 weeks (59%). In maternal rhesus D [Rh(D)] immunization 89% of fetuses survived, whereas survival in the case of Kell immunization was 58%. All fetuses with anemia due to Rh(c) immunization survived. The overall fatal procedure-related complication rate was 1.7% per procedure, resulting in a fetal loss rate of 4.8%., Conclusions: Intrauterine intravascular transfusions are effective in the management of fetal alloimmune anemia. Fetal hydrops, mostly associated with late referral, decreases the chance of survival. To improve the outcome of red-cell alloimmunized pregnancies early diagnosis of fetal anemia and referral to a specialized center are important, enabling the start of treatment before hydrops develops.

Maternal anti-HPA-1a antibodies can cause severe fetal and neonatal alloimmune thrombocytopenia (FNAIT), complicated by intracranial haemorrhage (ICH). Antenatal treatment with maternal intravenous immunoglobulin (IVIG) seems to protect against ICH even when thrombocytopenia persists. The aim of this study was to investigate if anti-HPA-1a antibodies and IVIG potentially affect vascular endothelial cells (ECs) in order to identify susceptibility for ICH. Human umbilical cord endothelial cells (HUVEC) were incubated with anti-HPA-1a antibodies with or without polyclonal IVIG and evaluated for EC activation. Maternal sera with anti-HPA-1a antibodies affected neither the EC expression of intracellular adhesion molecule-1 (ICAM-1), vascular adhesion molecule-1 (VCAM-1) and tissue factor (TF) nor the release of van Willebrand factor (vWF) or interleukin (IL)-8 nor the integrity of ECs. Maternal sera obtained after IVIG treatment and polyclonal IVIG decrease constitutive and cytokine-induced ICAM-1 and VCAM-1 expression on ECs. The results show that maternal anti-HPA-1a antibodies cause no activation or damage of ECs in this model. The clinical relevance of the de-activating properties of IVIG on EC activation with respect to ICH deserves further investigation.

Objective: Evaluation of the initial results of selective fetoscopic laser coagulation of placental blood vessels for severe second-trimester twin-to-twin transfusion syndrome in the Netherlands., Design: Prospective cohort study., Method: Between August 2000 and December 31, 2002, 49 patients were included with severe second-trimester twin-to-twin transfusion syndrome. Treatment took place in the Leiden University Medical Centre and consisted of selective fetoscopic laser coagulation of anastomoses along the vascular equator on the placental surface. Follow-up was complete until six weeks after delivery., Results: Median gestational age was 20 weeks at fetoscopy and 34 weeks at birth. Perinatal survival rate was 65% (64/98). The treatment resulted in at least one surviving child in 80% of pregnancies., Conclusion: The results of fetoscopic laser treatment for severe second-trimester twin-to-twin transfusion syndrome in the Netherlands are similar to the recent results in specialised foreign centres.

The aim of this study was to quantify and compare leukocyte populations in term decidua basalis and parietalis obtained after spontaneous vaginal delivery (SVD) or elective cesarean section (CS) without labor. Decidua basalis and parietalis samples were obtained from placentas after SVD (n = 20) and after CS (n = 30). Following mechanical disaggregation, leukocytes were purified and stained with monoclonal antibodies. Percentages of leukocyte subclasses within the CD45(+) cell fraction and activated T cells were determined by flow cytometry. No differences were found in the percentages of CD45(+) cells or CD56(bright)CD16(-) uterine natural killer (NK) cells between decidua basalis from SVD and CS or between decidua parietalis from SVD and CS. In decidua basalis and parietalis from SVD, a significantly higher number of CD56(dim)CD16(+) NK cells was found compared to CS. In decidua basalis from SVD, there was a significantly lower percentage of CD14(+) cells and higher percentage of CD19(+) cells compared to CS. The percentage of CD3(+) T cells expressing CD25 or human leukocyte antigen (HLA)-DR was significantly decreased in decidua basalis and parietalis from SVD compared to CS. Comparison of decidua collected after SVD or CS suggests that labor is associated with dynamic changes in the distribution of decidual leukocytes, specifically NK and T cell subpopulations. In particular, the disappearance of the CD4(+)CD25(+) T cell population, which possibly contains a subpopulation of regulatory T cells, may contribute to the initiation of labor. Further investigation into factors affecting decidual leukocytes may expand our understanding of the immunological events at the maternal-fetal interface.

We studied the relationship between bilirubin concentrations in amniotic fluid and fetal blood in 68 non-hydropic rhesus d-alloimmunized anemic fetuses at first blood sampling. In these alloimmunized fetuses, the amniotic fluid/fetal blood ratio for bilirubin decreased from 0.09 at 28 weeks to 0.05 at 33 weeks. In normal fetuses, amniotic fluid/fetal blood ratios for bilirubin, and for albumin, are in the same range and show a similar decrease during gestation. We conclude that amniotic fluid bilirubin concentration is determined, firstly, by fetal blood bilirubin concentration and, secondly, by the amniotic fluid/fetal blood ratio of albumin. Among five possible pathways bilirubin could take to build up a concentration in amniotic fluid (fetal kidneys, lungs, skin, bowel, membranes), the intramembranous pathway is the only one that is compatible with the amniotic fluid/fetal blood ratios for bilirubin that we found and must therefore be the most important.

Objective: To determine the opinion of Dutch gynaecologists and registrars on caesarean section (CS) on request., Study Design: Anonymous postal survey., Methods: A structured survey was send to all 900 gynaecologists and registrars in The Netherlands. They were asked to what extent they were willing to accept a request for an elective caesarean section, without evident medical reason. The survey contained eight simulated cases in which the reason for this request differed (obstetrical history and course of the present pregnancy). In two cases, there was no medical indication at all to perform a caesarean section; and in a third case caesarean section was due to excessive maternal weight relatively contraindicated., Results: The response rate was 65%. Willingness to perform an elective caesarean section ranged from 17 to 81% between the cases. Main reasons to perform a caesarean section were: (a). autonomy; (b). an unfavourable course of delivery in the absence of motivation for a natural childbirth; (c). litigation. The main reasons to refuse a request for a caesarean section were: (a). higher maternal morbidity and mortality; (b). no indication for caesarean section. A logistic regression analysis on personal characteristics showed that an experienced doctor is more willing to perform an elective caesarean section then a consultant or registrar with limited experience. The sex of the doctor was of no influence and the same held for the University at which they had been trained. Furthermore, it seems that doctors are more willing to accept the request if it is based upon unfounded, but understandable fear., Conclusion: In The Netherlands, a woman can always find a gynaecologist willing to perform a caesarean section for non medical reasons. This willingness increases with the age of the doctor. There is a need for guidelines when handling these cases.