Vesicular uptake of macromolecules by human placental amniotic epithelial cells.

INTRODUCTION: Studies in animal models have shown that unidirectional vesicular transport of amniotic fluid across the amnion plays a primary role in regulating amniotic fluid volume. Our objective was to explore vesicle type, vesicular uptake and intracellular distribution of vesicles in human amnion cells using high- and super-resolution fluorescence microscopy. METHODS: Placental amnion was obtained at cesarean section and amnion cells were prepared and cultured. At 20%-50% confluence, the cells were incubated with fluorophore conjugated macromolecules for 1-30 min at 22 °C or 37 °C. Fluorophore labeled macromolecules were selected as markers of receptor-mediated caveolar and clathrin-coated vesicular uptake as well as non-specific endocytosis. After fluorophore treatment, the cells were fixed, imaged and vesicles counted using Imaris® software. RESULTS: Vesicular uptake displayed first order saturation kinetics with half saturation times averaging 1.3 min at 37 °C compared to 4.9 min at 22 °C, with non-specific endocytotic uptake being more rapid at both temperatures. There was extensive cell-to-cell variability in uptake rate. Under super-resolution microscopy, the pattern of intracellular spatial distribution was distinct for each macromolecule. Co-localization of fluorescently labeled macromolecules was very low at vesicular dimensions. CONCLUSIONS: In human placental amnion cells, 1) vesicular uptake of macromolecules is rapid, consistent with the concept that vesicular transcytosis across the amnion plays a role in the regulation of amniotic fluid volume; 2) uptake is temperature dependent and variable among individual cells; 3) the unique intracellular distributions suggest distinct functions for each vesicle type; 4) non-receptor mediated vesicular uptake may be a primary vesicular uptake mechanism.

Maternal co-morbidities and neonatal outcomes associated with cystic fibrosis.

OBJECTIVE: To evaluate maternal co-morbidities and adverse perinatal outcomes associated with cystic fibrosis (CF). METHODS: This is a retrospective cohort study of 2 178 954 singleton pregnancies at ≥20 weeks' gestation with and without CF in the state of California during the years 2005-2008. ICD-9 codes and linked hospital discharge and vital statistics data were utilized. Rates of maternal co-morbidities, fetal congenital anomalies and adverse perinatal outcomes were compared in those with CF and those without. Maternal co-morbidities included gestational hypertension, preeclampsia, gestational diabetes and primary cesarean delivery. Perinatal outcomes included neonatal demise, preterm birth, intrauterine growth restriction, macrosomia, anomaly, fetal demise, asphyxia, respiratory distress syndrome, jaundice, intraventricular hemorrhage, hypoglycemia and necrotizing enterocolitis. RESULTS: The cohort included 2 178 954 pregnancies of which 77 mothers had CF. Mothers with CF were more likely to have pre-gestational diabetes and had higher rates of primary cesarean delivery. Neonates delivered to mothers with CF were more likely to be born preterm and have congenital anomalies but otherwise were not at increased risk for significant neonatal morbidity or mortality when adjusted for gestational age. CONCLUSION: Mothers with CF are more likely to have pre-gestational diabetes, deliver preterm (<37 weeks gestation) and have a primary cesarean delivery. Infants are more likely to have congenital anomalies. In addition to early diabetic screening and genetic counseling, a detailed fetal anatomy ultrasound should be performed in women with CF.

Correlation of serum fructosamine and recurrent pregnancy loss: Case-control study.

AIM: Pre-gestational diabetes is associated with an elevated risk of pregnancy loss, but it is unclear whether subclinical glucose intolerance is associated with pregnancy loss, especially recurrent pregnancy loss (RPL). The aim of this study was therefore to compare maternal serum fructosamine (a marker of glycemic control) in patients with and without RPL. METHODS: A case-control study was carried out of 117 women with unexplained RPL, defined as two or more pregnancy losses with no more than one live birth, and 117 age-matched controls with at least one full-term uncomplicated pregnancy and no more than one pregnancy loss. No RPL patients or controls had a clinical diagnosis of pre-gestational or gestational diabetes. Maternal serum was analyzed for fructosamine on quantitative spectrophotometry. RESULTS: Mean body mass index (BMI) of RPL patients was 26.0 ± 6.4 kg/m(2) compared with 26.6 ± 5.8 kg/m(2) (P = 0.40). Fructosamine was higher in women with RPL (224.1 ± 28.79 µmol/mL) compared with controls (188.9 ± 19.3 µmol/mL, P < 0.001). This difference persisted when RPL patients and controls were stratified by BMI. The proportion of women with elevated fructosamine considered diagnostic of diabetes (>285 µmol/L) was similar in RPL patients and controls. CONCLUSION: The RPL patients and controls had a similar proportion of women with elevated fructosamine considered diagnostic of diabetes. Serum fructosamine was increased in women with RPL compared with controls. Thus, subclinical glucose intolerance may be associated with an increased risk of RPL. These data support further investigation into the mechanisms of RPL associated with glucose intolerance, but do not support testing for subclinical glucose intolerance in women with RPL.

Fetal malposition: impact and management.

Fetal malposition, either occiput posterior or transverse (OT), leads to greater risk of cesarean delivery, prolonged labor, and increased perinatal morbidity. Historically, there is a known association between epidural use and malposition that was assumed to be due to the increased discomfort of laboring with a fetus in the occiput posterior position. However, evidence now suggests that the epidural itself may contribute to fetal malposition by impacting the probability of internal rotation. Fetal malposition may be impacted by manual rotation. Manual rotation has been associated with greater rates of delivering in the occiput anterior position and lower rates of cesarean delivery.

Management of fetal malpresentation.

Fetal malpresentation is an important cause of the high cesarean delivery rate in the United States and around the world. This includes breech, face, brow, and compound presentations as well as transverse lie. Risk factors include multiparity, previously affected pregnancy, polyhydramnios, and fetal and uterine anomalies. Appropriate management can reduce the need for cesarean delivery in some cases. This review discusses management options and focuses specifically on external cephalic version and vaginal breech delivery.

Celiac disease serum markers and recurrent pregnancy loss.

Celiac disease has been associated with numerous unfavorable health outcomes, including pregnancy complications such as infertility, preterm birth, and preeclampsia. However, the association between celiac disease and recurrent pregnancy loss (RPL) remains uncertain. Our purpose was to compare serum markers of celiac disease in women with and without RPL. Therefore, we performed a case-control study of 116 women with unexplained recurrent pregnancy loss and 116 age-matched controls. Maternal sera were analyzed for immunoglobulin A (IgA) and immunoglobulin G (IgG) tissue transglutaminase (tTG) antibodies and endomysial (EM) antibodies. Groups were similar with regard to age, race and ethnicity, and BMI. One case and one control tested positive (≥20 Units) for IgA tTG antibodies and mean levels of IgA tTG antibodies were similar in cases and controls (5.5±2.86 versus 6.0±12.45; p=0.16). No cases or controls were positive for IgG tTG antibodies. However, cases had higher levels of IgG tTG antibody compared with controls (4.0±2.40 versus 3.3±1.30; p=0.0064). One subject (a control) tested positive for IgA EM antibodies and no subjects tested positive for IgG EM antibodies. In conclusion, positive results for tTG and EM antibodies were similar in women with and without RPL. Given these results, testing for occult celiac disease is not recommended in the evaluation of women with idiopathic RPL.

Staged cleft sternum repair using dermal allograft and synthetic mesh.

Cleft sternum is a rare congenital chest deformity that develops during the first trimester. Failure of the process of midline mesenchymal strip fusion leads to absence of the sternum, resulting in cleft formation. Multiple surgical approaches have been described in the closure of sternal clefts. An optimal surgical approach is still debatable. We describe 2 cases of complete sternal clefting treated with staging of the repair. Dermal allograft and synthetic mesh along with myofasciocutaneous flaps are used a bridging method to future definitive treatment. Most patients will require secondary cardiothoracic procedures for underlying cardiac conditions, and disruption of any primary repair is compromised on reentry into the chest. Staging this procedure avoids this potential problem. Also, concerns regarding chest wall constriction and cardiopulmonary compromise are minimized. Once the child has matured, definitive treatment can be pursued with more abundant autologous donor tissue. Our approach is safe with minimal complications and is well tolerated by the patients.