The adipokine FABP4 is a key regulator of neonatal glucose homeostasis.

During pregnancy, fetal glucose production is suppressed, with rapid activation immediately postpartum. Fatty acid-binding protein 4 (FABP4) was recently demonstrated as a regulator of hepatic glucose production and systemic metabolism in animal models. Here, we studied the role of FABP4 in regulating neonatal glucose hemostasis. Serum samples were collected from pregnant women with normoglycemia or gestational diabetes at term, from the umbilical circulation, and from the newborns within 6 hours of life. The level of FABP4 was higher in the fetal versus maternal circulation, with a further rise in neonates after birth of approximately 3-fold. Neonatal FABP4 inversely correlated with blood glucose, with an approximately 10-fold increase of FABP4 in hypoglycemic neonates. When studied in mice, blood glucose of 12-hour-old WT, Fabp4-/+, and Fabp4-/- littermate mice was 59 ± 13 mg/dL, 50 ± 11 mg/dL, and 43 ± 11 mg/dL, respectively. Similar to our observations in humans, FABP4 levels in WT mouse neonates were approximately 8-fold higher compared with those in adult mice. RNA sequencing of the neonatal liver suggested altered expression of multiple glucagon-regulated pathways in Fabp4-/- mice. Indeed, Fabp4-/- liver glycogen was inappropriately intact, despite a marked hypoglycemia, with rapid restoration of normoglycemia upon injection of recombinant FABP4. Our data suggest an important biological role for the adipokine FABP4 in the orchestrated regulation of postnatal glucose metabolism.

Maternal Therapy with Ad.VEGF-A165 Increases Fetal Weight at Term in a Guinea-Pig Model of Fetal Growth Restriction.

In a model of growth-restricted sheep pregnancy, it was previously demonstrated that transient uterine artery VEGF overexpression can improve fetal growth. This approach was tested in guinea-pig pregnancies, where placental physiology is more similar to humans. Fetal growth restriction (FGR) was attained through peri-conceptual nutrient restriction in virgin guinea pigs. Ad.VEGF-A165 or Ad.LacZ (1 × 1010vp) was applied at mid-gestation via laparotomy, delivered externally to the uterine circulation with thermosensitive gel. At short-term (3-8 days post surgery) or at term gestation, pups were weighed, and tissues were sampled for vector spread analysis, VEGF expression, and its downstream effects. Fetal weight at term was increased (88.01 ± 13.36 g; n = 26) in Ad.VEGF-A165-treated animals compared with Ad.LacZ-treated animals (85.52 ± 13.00 g; n = 19; p = 0.028). The brain, liver, and lung weight and crown rump length were significantly larger in short-term analyses, as well as VEGF expression in transduced tissues. At term, molecular analyses confirmed the presence of VEGF transgene in target tissues but not in fetal samples. Tissue histology analysis and blood biochemistry/hematological examination were comparable with controls. Uterine artery relaxation in Ad.VEGF-A165-treated dams was higher compared with Ad.LacZ-treated dams. Maternal uterine artery Ad.VEGF-A165 increases fetal growth velocity and term fetal weight in growth-restricted guinea-pig pregnancy.

Gene Targeting to the Uteroplacental Circulation of Pregnant Guinea Pigs.

Our study aimed to target adenoviral gene therapy to the uteroplacental circulation of pregnant guinea pigs in order to develop a novel therapy for fetal growth restriction. Four methods of delivery of an adenovirus encoding ß-galactosidase (Ad.LacZ) were evaluated: intravascular injection using phosphate-buffered saline (PBS) into (1) uterine artery (UtA) or (2) internal iliac artery or external administration in (3) PBS or (4) pluronic F-127 gel (Sigma Aldrich). Postmortem examination was performed 4 to 7 days after gene transfer. Tissue transduction was assessed by X-gal histochemistry and enzyme-linked immunosorbent assay. External vascular application of the adenovirus vector in combination with pluronic gel had 91.7% success rate in terms of administration (85% maternal survival) and gave the best results for maternal/fetal survival and local transduction efficiency without any spread to maternal or fetal tissues. This study suggests an optimal method of gene delivery to the UtAs of a small rodent for preclinical studies.

Induction of labor for term small-for-gestational-age fetuses: what are the consequences?

OBJECTIVE: To evaluate whether early term labor induction for suspected intrauterine growth restriction (weeks 37-39) improves neonatal outcome for small-for-gestational-age (SGA) neonates. STUDY DESIGN: Delivery room data for 2004-2008 from a single tertiary medical center were linked to neonatal discharge data from the same institution. Data were limited to all singleton, liveborn SGA neonates born at 37-42 weeks of gestation and their mothers. Births with known congenital anomalies were excluded. Women undergoing induction of labor for suspected growth restriction between 37 and 39 weeks' gestation (early induction SGA) were compared with women who gave birth to term SGA neonates without early induction. SGA (<10th percentile for gestational age and gender) was used as a surrogate for intrauterine growth restriction. Associations between early term labor induction and neonatal morbidities were estimated using logistic regression. RESULTS: A total of 2378 SGA neonates meeting study criteria were identified. Of these, 445 underwent early term induction and 1933 were in the non-early induction SGA group. Intrauterine demise among term (37-42 weeks) SGAs occurred in one case at 37 weeks. Early term induction for SGA was associated with an increased risk of cesarean delivery. Several neonatal complications, including hyperbilirubinemia, hypoglycemia and respiratory complications were more prevalent in the early induction SGA group. The increased odds for neonatal complications persisted after controlling for possible confounders. CONCLUSIONS: Early term induction for SGA fetuses results in an increased risk of cesarean deliveries as well as neonatal metabolic and respiratory complications, with no apparent neonatal benefit.

Subsequent pregnancy after stillbirth: obstetrical and medical risks.

OBJECTIVE: To evaluate obstetric outcome after stillbirth according to placental and prothrombotic risk factors. METHODS: Obstetric outcomes of women with prior stillbirth and subsequent pregnancies were reviewed. Data on the immediate subsequent pregnancy included fetal loss, stillbirth, obstetric/medical complications, gestational age and birth weight at delivery, mode of delivery, thrombophilia, and prescribed medication. Placental stillbirth was defined as stillbirth associated with placental abruption, intrauterine growth restriction (IUGR), or histological evidence of placental infarcts. Controls were unselected women who gave birth at our center during a single calendar year. Factors influencing recurrence risks were estimated. RESULTS: Seventy-three subsequent pregnancies were identified. Five out of 73 (6.8%) women had a repeat stillbirth, significantly higher than controls (relative risk 22.2, 95% confidence interval 8.9-55.4). Four out of five repeat stillbirth cases occurred <37 weeks gestation. Hypertensive complications, diabetes and abruption were higher, while gestational age and birth weight at delivery were significantly lower than controls. Prior placental stillbirth was associated with a 10.5 times higher risk of IUGR in the subsequent pregnancy compared with non-placental stillbirth. All five repeat stillbirth cases occurred in thrombophilic women. CONCLUSION: Women with prior stillbirth face an increased risk of pregnancy complications and stillbirth recurrence, especially with concurrent thrombophilia. Most repeat stillbirth cases occur preterm.

Thrombophilic risk factors for placental stillbirth.

OBJECTIVES: To define the characteristics of placental stillbirth and the possible contribution of thrombophilic risk factors. STUDY DESIGN: A prospective cohort study was performed. Women diagnosed with antenatal stillbirth (>20 weeks) of singleton pregnancies between 2006 and 2008 were referred postpartum for evaluation. Maternal risk factors, fetal, placental and cord abnormalities, and a detailed thrombophilia screening, including inherited and acquired thrombophilia, were evaluated. Fetal autopsy and placental pathology were encouraged. Placental stillbirth was defined as death of a normally-formed fetus with evidence of intrauterine fetal growth restriction, oligohydramnios, placental abruption and/or histological evidence of placental contribution to fetal death. Pregnancy characteristics and thrombophilia profiles were compared between placental and non-placental stillbirth cases. RESULTS: Sixty-seven women with stillbirth comprised the study group. Placental stillbirth was evident in 33/67 (49.3%). Significantly more women with placental stillbirth were nulliparous, when compared with non-placental stillbirth women (21/33 vs. 9/34, p=0.002). Mean gestational age was lower for placental, compared with non-placental stillbirth (31.1 ± 6.1 weeks vs. 33.9 ± 4.8 weeks, p=0.04), as was birth weight. Thirty-six of the 67 women (53.7%) tested positive for at least one thrombophilia. The prevalence of maternal thrombophilia was higher for placental stillbirth women (63.6%), and even higher (69.6%) for women after preterm (<37 weeks) placental stillbirth. Factor V Leiden and/or prothrombin G20210A mutation were much more prevalent in placental versus non-placental stillbirth women (OR 3.06, 95% CI 1.07-8.7). CONCLUSIONS: Placental stillbirth comprises a unique subgroup with specific maternal characteristics. Maternal thrombophilia is highly prevalent, especially in preterm placental stillbirth. This may have implications for the management strategy in future pregnancies in this subgroup.

Production of bioactive, post-translationally modified, heterotrimeric, human recombinant type-I collagen in transgenic tobacco.

Collagen's biocompatibility, biodegradability and low immunogenicity render it advantageous for extensive application in pharmaceutical or biotechnological disciplines. However, typical collagen extraction from animal or cadaver sources harbors risks including allergenicity and potential sample contamination with pathogens. In this work, two human genes encoding recombinant heterotrimeric collagen type I (rhCOL1) were successfully coexpressed in tobacco plants with the human prolyl-4-hydroxylase (P4H) and lysyl hydroxylase 3 (LH3) enzymes, responsible for key posttranslational modifications of collagen. Plants coexpressing all five vacuole-targeted proteins generated intact procollagen yields of approximately 2% of the extracted total soluble proteins. Plant-extracted rhCOL1 formed thermally stable triple helical structures and demonstrated biofunctionality similar to human tissue-derived collagen supporting binding and proliferation of adult peripheral blood-derived endothelial progenitor-like cells. Through a simple, safe and scalable method of rhCOL1 production and purification from tobacco plants, this work broadens the potential applications of human recombinant collagen in regenerative medicine.

Versatile protein microarray based on carbohydrate-binding modules.

Non-DNA microarrays, such as protein, peptide and small molecule microarrays, can potentially revolutionize the high-throughput screening tools currently used in basic and pharmaceutical research. However, fundamental obstacles remain that limit their rapid and widespread implementation as an alternative bioanalytical approach. These include the prerequisite for numerous proteins in active and purified form, ineffectual immobilization strategies and inadequate means for quality control of the considerable numbers of multiple reagents. This study describes a simple yet efficient strategy for the production of non-DNA microarrays, based on the tenacious affinity of a carbohydrate-binding module (CBM) for its three-dimensional substrate, i.e., cellulose. Various microarray formats are described, e.g., conventional and single-chain antibody microarrays and peptide microarrays for serodiagnosis of human immunodeficiency virus patients. CBM-based microarray technology overcomes many of the previous obstacles that have hindered fabrication of non-DNA microarrays and provides a technically simple but effective alternative to conventional microarray technology.

Uterine rupture: differences between a scarred and an unscarred uterus.

OBJECTIVES: This study aimed to compare risk factors, site of rupture, and outcome of uterine rupture among patients with a scarred versus an unscarred uterus. Study design We conducted a comparison between all cases of uterine rupture (n=53) in women with a scarred versus an unscarred uterus, occurring between January 1988 and July 2002. RESULTS: During the study period, there were ruptures among 26 patients with a scarred uterus and 27 patients without a uterine scar. No significant differences were noted between the scarred and unscarred groups while comparing risk factors such as birth order, birth weight, hydramnios, oxytocin induction, diabetes, and malpresentation. The main site of involvement in both groups was the lower uterine segment representing 92.6% of the ruptures in the unscarred group and 92.3% of the ruptures in the scarred uterus group. Cervical involvement was significantly more common among patients without a previous uterine scar (33.3% vs 7.7%; odds ratio [OR]=6.0, 95% CI, 1.16-31.23, P=.04). Conversely, uterus corpus involvement did not differ between the groups. Perinatal mortality did not differ between the groups. In addition, no significant differences were noted regarding maternal morbidity such as the need for hysterectomy, blood transfusion, or length of hospitalization. CONCLUSION: Although cervical involvement was significantly more prevalent in the rupture of an unscarred uterus, no significant differences in maternal or perinatal morbidity were noted between rupture of a scarred versus an unscarred uterus.

Changes in fetal heart rate and uterine patterns associated with uterine rupture.

OBJECTIVE: To determine changes in fetal heart rate (FHR) and uterine patterns preceding complete uterine rupture. STUDY DESIGN: FHR and uterine patterns of 50 women with uterine rupture were compared with 601 tracings of controls without scarred uteri. Tracings were interpreted using the National Institute of Child Health and Human Development Research Planning Workshop guidelines. RESULTS: Interobserver and intraobserver agreements of FHR and uterine tracings in the uterine rupture group were excellent (kappa of .96 for both variables). Comparing tracing patterns during the first stage, higher rates of severe fetal bradycardia (4.0% vs. 1.0%, P = .064), fetal tachycardia (8.0% vs. 2.3%, P = .042), reduced baseline variability (24.0% vs. 12.5%, P = .021), uterine tachysystole (10.0% vs. 0.8%, P < .001) and disappearance of contractions (6.0% vs. 0, P < .001) were noted among patients with uterine rupture as compared to the controls. During the second stage of labor, patients with uterine rupture had significantly higher rates of reduced baseline variability (47.8% vs. 7.7%, P < .001), severe variable decelerations (26.1% vs. 6.4%, P = .004), uterine tachysystole (22.0% vs. 0.5%, P < .001) and disappearance of contractions (13.0% vs. 0, P < .001). Using a backward, stepwise multiple logistic regression model, severe fetal bradycardia (OR = 8.2, 95% CI 2.2-31.0, P = .002) and uterine tachysystole (OR = should alert the 8.0, 95% CI 1.7-37.9, P = .008) were found to be independent patterns preceding uterine rupture during the first stage of labor. Likewise, during the second stage, reduced baseline variability (OR = 4.2, 95% CI 1.4-12.3, P = .009) and uterine tachysystole (OR = 42.3, 95% CI 10.6-168.3, P < .001) were independently associated with uterine rupture in another multivariable analysis. CONCLUSION: Abnormal monitor patterns among women presenting with risk factors for uterine rupture, specifically uterine tachysystole, reduced baseline variability and severe bradycardia, should act as warning signs to the obstetrician.

Uterine rupture: risk factors and pregnancy outcome.

OBJECTIVES: This study aimed at determining risk factors and pregnancy outcome in women with uterine rupture. STUDY DESIGN: We conducted a population-based study, comparing all singleton deliveries with and without uterine rupture between 1988 and 1999. RESULTS: Uterus rupture occurred in 0.035% (n=42) of all deliveries included in the study (n=117,685). Independent risk factors for uterine rupture in a multivariable analysis were as follows: previous cesarean section (odds ratio [OR]=6.0, 95% CI 3.2-11.4), malpresentation (OR=5.4, 95% CI 2.7-10.5), and dystocia during the second stage of labor (OR=13.7, 95% CI 6.4-29.3). Women with uterine rupture had more episodes of postpartum hemorrhage (50.0% vs 0.4%, P<.01), received more packed cell transfusions (54.8% vs 1.5%, P<.01), and required more hysterectomies (26.2% vs 0.04%, P<.01). Newborn infants delivered after uterine rupture were more frequently graded Apgar scores lower than 5 at 5 minutes and had higher rates of perinatal mortality when compared with those without rupture (10.3% vs 0.3%, P<.01; 19.0% vs 1.4%, P<.01, respectively). CONCLUSION: Uterine rupture, associated with previous cesarean section, malpresentation, and second-stage dystocia, is a major risk factor for maternal morbidity and neonatal mortality. Thus, a repeated cesarean delivery should be considered among parturients with a previous uterine scar, whose labor failed to progress.