March Of Dimes Foundation Research Articles

Magnesium levels in relation to rates of preterm birth: a systematic review and meta-analysis of ecological, observational, and interventional studies.

Experimental studies suggest that magnesium levels in pregnant women may affect the length of gestation, as magnesium affects the activity of smooth muscle in the uterus. Little is known about the association between magnesium levels or supplementation and the rate of preterm birth. The aim of this systematic review was to summarize the data on magnesium soil levels and preterm birth rates from ecological, observational, and interventional studies. Soil magnesium levels were obtained from US Geological Survey data, and preterm birth rates were acquired from the March of Dimes Foundation. Relevant epidemiological and clinical studies published until April 2019 in peer-reviewed journals were retrieved from PubMed, Google Scholar, and related reference lists. Original studies published in English, conducted in humans, and in which magnesium (dietary/supplemental intake or biomarkers) was an exposure and preterm birth was an outcome were included. Eleven studies were included in the systematic review. Meta-analysis was performed on 6 studies. Overall relative risk (RR) and corresponding 95%CIs for risk of preterm birth in relation to magnesium supplementation were estimated by a random-effects model. The ecological study revealed an inverse correlation between magnesium content in soil and rates of preterm birth across the United States (r = -0.68; P < 0.001). Findings from 11 observational studies generally support an inverse association between serum magnesium levels and rates of preterm birth. Of the 6 eligible randomized controlled trials, which included 3068 pregnant women aged 20 to 35 years and 352 preterm infants, the pooled RR was 0.58 (95%CI, 0.35-0.96) for women in the magnesium supplementation group compared with women in the control group. Accumulated evidence from ecological, observational, and interventional studies consistently indicates that adequate magnesium intake during pregnancy may help reduce the incidence of preterm birth.

Structure and mechanism of DNMT3A‐mediated DNA methylation

Mammalian DNA methylation is an important epigenetic mechanism crucial for gene silencing and imprinting, X‐inactivation, genome stability, and cell fate determination. It is established mainly at CpG dinucleotides by de novo methyltransferases DNMT3A and DNMT3B, and subsequently maintained by DNA methyltransferase 1 (DNMT1) in a replication‐dependent manner. Deregulation of this process is implicated in various diseases, notably cancer. However, the mechanisms underlying DNMT3 substrate recognition and enzymatic specificity remain elusive. We determined the crystal structure of the DNMT3A – DNMT3L tetramer in complex with a DNA substrate containing two target sites separated by fourteen base pairs. Structural analysis of the DNMT3A – DNMT3L – DNA complex reveals that the two DNMT3A monomers simultaneously attack the two target sites, supporting the notion that DNMT3A homodimer can co‐methylate two adjacent CpG dinucleotides in one DNA‐binding event. Importantly, we identified that a residue from the target recognition domain makes crucial contacts with CpG, ensuring DNMT3A enzymatic preference towards CpG sites in cells. Mutational, enzymatic and cellular analyses further demonstrate that the hematological cancer‐associated somatic mutations of the substrate‐binding residues decrease DNMT3A activity, induce CpG hypomethylation, and promote transformation of hematopoietic cells. Together, our study reveals the mechanistic basis for DNMT3A‐mediated DNA methylation.Support or Funding InformationSidney Kimmel Foundation for Cancer Research;March of Dimes Foundation (1‐FY15‐345);National Institute of Health (1R35GM119721)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Opioid Use in Pregnancy, Neonatal Abstinence Syndrome, and Childhood Outcomes: Executive Summary of a Joint Workshop by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, American College of Obstetricians and Gynecologists, American Academy of Pediatrics, Society for Maternal-Fetal Medicine, Centers for Disease Control and Prevention, and the March of Dimes Foundation.

Opioid Use in Pregnancy, Neonatal Abstinence Syndrome, and Childhood Outcomes: Executive Summary of a Joint Workshop by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, American College of Obstetricians and Gynecologists, American Academy of Pediatrics, Society for Maternal-Fetal Medicine, Centers for Disease Control and Prevention, and the March of Dimes Foundation.

Opioid Use in Pregnancy, Neonatal Abstinence Syndrome, and Childhood Outcomes: Executive Summary of a Joint Workshop by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, American College of Obstetricians and Gynecologists, American Academy of Pediatrics, Society for Maternal-Fetal Medicine, Centers for Disease Control and Prevention, and the March of Dimes Foundation

(Abstracted from Obstet Gynecol 2017;130(1):10–28) This article provides a summary of the “Opioid Use in Pregnancy, Neonatal Abstinence Syndrome, and Childhood Outcomes” workshop convened by the Eunice Kennedy Shriver National Institute of Child Health and Human Development on April 4 and 5, 2016, and cosponsored by the American College of Obstetricians and Gynecologists (ACOG), the American Academy of Pediatrics, the Society for Maternal-Fetal Medicine, the Centers for Disease Control and Prevention, and the March of Dimes. Expert speakers from diverse fields and workshop participants identified scientific opportunities to advance the understanding of opioid use disorders in pregnancy and to improve outcomes for affected women, their children, and their families.

Building Bridges for Collaboration: the University of Iowa Women's Health Tissue Repository

The University of Iowa Women's Health Tissue Repository (WHTR) is a large biobank in which specimens collected and have significant clinical annotations with the goal of facilitating research. This umbrella organization contains the Maternal Fetal Tissue Bank (IRB#200910874), Paternal Contributions to Children's Health (IRB# 201208773), Reproductive, Endocrinology and Infertility Tissue Bank (IRB# 201010769), Well Woman Bank (IRB# 201202714), and Gynecologic Malignancies Tissue Bank (IRB# 200209010). Patient consent and sample collection, processing, and storage is carried out by the Department of Obstetrics &amp; Gynecology.In 2013, the University of Iowa instituted the UI Bioshare program and it went live in 2015. UI Bioshare is an enterprise‐level biospecimen management system. This system established a common ontology for use across biobanks at our institution. Goals of the program include 1) users being able to explore aggregate inventory queries, 2) to develop resources to decrease manual data abstraction and entry and 3) to position the University of Iowa for large‐scale projects in clinical research and personalized medicine initiatives. A core group of biobanks, including the Women's Health Tissue Repository, participated in the purchase, design, and implementation of the program.As a result of the implementation, the WHTR now shares a common data dictionary with our institutional peer biobanks. The common definitions have allowed for an honest broker in UI Bioshare to query our databases to gather aggregate information for grant submissions. Our sample inventory management is significantly improved for data entry and for queries by investigators. In total, 180,000 samples were migrated into UI Bioshare and 200,000 have been newly entered since the go live. Of these 55,000 samples were migrated for the WHTR and 15,750 have been collected and entered by us since our go‐live in September of 2015. At the enterprise level, information form 18,000 unique participants were migrated and 3000 new participants have been added. The WHTR migrated information from 2700 participants and have enrolled 750 new participants. We have also experienced a reduction in manual data extraction from the electronic medical record. This has significantly reduced the time required to extract information such as demographics from weeks to minutes. Users of the WHTR have been able to perform queries of samples and data independently; the amount of information available to them is based on permissions and roles built into the software. We now have collaborations with other departments including Pharmacology, Health &amp; Human Physiology, Pediatrics, Toxicology, and Epidemiology. In addition, we collaborate with other groups within the US and internationally.Despite initial concerns regarding data ownership, changes to workflow, and the time investment required for migrating data, the experience of the Women's Health Tissue Repository in participating in an enterprise‐wide system has been positive. Our biorepository is no longer a silo of information, but is now bridged to other university biobanks which allows for larger‐scale collaborations.Support or Funding InformationUniversity of Iowa Carver College of Medicine, Holden Comprehensive Cancer Center, National Institutes of Health CTSA U54TR001356, Department of Obstetrics &amp; Gynecology, American Heart Association SFRN in Hypertension, Reproductive Scientist Development Program (NIH K12 HD000849), Burroughs Wellcome Fund (1015358), March of Dimes Foundation (4‐FY15‐415)

Antiphospholipid antibody-induced miR-146a-3p drives trophoblast interleukin-8 secretion through activation of Toll-like receptor 8.

What is the role of microRNAs (miRs) in antiphospholipid antibody (aPL)-induced trophoblast inflammation? aPL-induced up-regulation of trophoblast miR-146a-3p is mediated by Toll-like receptor 4 (TLR4), and miR-146a-3p in turn drives the cells to secrete interleukin (IL)-8 by activating the RNA sensor, TLR8. Obstetric antiphospholipid syndrome (APS) is an autoimmune disorder characterized by circulating aPL and an increased risk of pregnancy complications. We previously showed that aPL recognizing beta2 glycoprotein I (β2GPI) elicit human first trimester trophoblast secretion of IL-8 by activating TLR4. Since some miRs control TLR responses, their regulation in trophoblast cells by aPL and functional role in the aPL-mediated inflammatory response was investigated. miRs can be released from cells via exosomes, and therefore, miR exosome expression was also examined. A panel of miRs was selected based on their involvement with TLR signaling: miR-9; miR-146a-5p and its isomiR, miR-146a-3p; miR-155, miR-210; and Let-7c. Since certain miRs can activate the RNA sensor, TLR8, this was also investigated. For in vitro studies, the human first trimester extravillous trophoblast cell line, HTR8 was studied. HTR8 cells transfected to express a TLR8 dominant negative (DN) were also used. Plasma was evaluated from pregnant women who have aPL, either with or without systemic lupus erythematous (SLE) (n = 39); SLE patients without aPL (n = 30); and healthy pregnant controls (n = 20). Trophoblast HTR8 wildtype and TLR8-DN cells were incubated with or without aPL (mouse anti-human β2GPI mAb) for 48-72 h. HTR8 cells were also treated with or without aPL in the presence and the absence of a TLR4 antagonist (lipopolysaccharide from Rhodobacter sphaeroides; LPS-RS), specific miR inhibitors or specific miR mimics. miR expression levels in trophoblast cells, trophoblast-derived exosomes and exosomes isolated from patient plasma were measured by qPCR. Trophoblast IL-8 secretion was measured by ELISA. aPL significantly increased trophoblast cellular and exosome expression of miR-146a-5p, miR-146a-3p, miR-155 and miR-210. aPL-induced up-regulation of trophoblast miR-146a-5p, miR-146a-3p and miR-210, but not miR-155, was inhibited by the TLR4 antagonist, LPS-RS. While inhibition or overexpression of miR-146a-5p had no effect on aPL-induced trophoblast IL-8 secretion, miR-146a-3p inhibition significantly reduced this response. aPL-induced trophoblast IL-8 secretion was inhibited by the presence of the TLR8-DN. In the absence of aPL, transfection of trophoblast cells with a miR-146a-3p mimic significantly increased IL-8 secretion and this was inhibited by the presence of the TLR8-DN. Patients with aPL and adverse pregnancy outcomes (APOs) expressed significantly higher levels of circulating miR-146a-3p compared with healthy pregnant controls with no pregnancy complications (P < 0.05). While the enrichment of miR-146a-3p in trophoblast-derived exosomes support the role of this miR acting in a paracrine or endocrine manner through exosome delivery, this has not been demonstrated. However, miR-146a-3p may also exert its pro-inflammatory effect intracellularly within the same trophoblast cell targeted by aPL. These findings provide a novel mechanism of trophoblast inflammation through miRs activating RNA-sensing receptors. Furthermore, circulating exosomal-associated miR-146a-3p in APS patients may serve clinically as a biomarker for related APOs. This study was supported in part by grants from the American Heart Association (#10GRNT3640032 to V.M.A.), the March of Dimes Foundation (Gene Discovery and Translational Research Grant #6-FY12-255 to V.M.A.), NICHD, NIH (R01HD049446 to V.M.A.), the Gina M. Finzi Memorial Student Summer Fellowship from the Lupus Foundation of America (to S.M.G.), and the Yale University School of Medicine Medical Student Fellowship (to S.M.G.). The authors declare no competing financial interests. N/A.

Peri-ovulatory putrescine supplementation reduces embryo resorption in older mice.

Does peri-ovulatory putrescine supplementation of older mice improve oocyte quality and reduce the incidence of embryo resorption? Peri-ovulatory putrescine supplementation in older mice improved oocyte quality, as indicated by increased blastocyst cell numbers and reduced the incidence of embryo resorption. Rodents exhibit a transient rise of ornithine decarboxylase (ODC) and putrescine in the ovaries during ovulation. Older mice exhibit reduced ovarian ODC activity during ovulation. Supplementation of in vitro maturation medium with putrescine reduces oocyte aneuploidy rates of older mice. The rationale was to correct ovarian putrescine deficiency in older mice by peri-ovulatory putrescine supplementation in drinking water and to observe the reproductive consequences of this intervention. This project was conducted between 2010 and 2014. Older mice (9-11 months of age) were given regular drinking water (control) or drinking water with 1% putrescine dihydrochloride (62 mM) for 2-4 days before mating. Plugged mice were then withdrawn from putrescine supplementation. Blastocysts were retrieved on 3.5 days post coitum (dpc) for the determination of cell numbers. For resorption analyses, mice were killed on 9.5 dp or 12.5 dpc, and implantation sites were dissected to determine the embryo status. For birth studies, mice were examined every morning between 16.5 and 23.5 dpc. Births were recorded as live or stillbirth. We demonstrated that deficiency of ovarian putrescine in older mice can be restored by peri-ovulatory putrescine supplementation in drinking water. Putrescine supplementation in older mice increased blastocyst cell numbers (from 40 to 54; P < 0.0001, t-test), reduced embryo resorption rates (from 41.1 to 15.4% in old C57BL/6 mice, P < 0.0001, Fisher's exact test; from 14.2 to 6.4% in old CF1 mice, P = 0.004, Fisher's exact test), and doubled the number of live born pups. Furthermore, exogenous putrescine exhibited rapid absorption and excretion, and showed no toxicity to mothers or fetuses. The mechanism of putrescine action in oocytes and/or ovaries remains unclear. Peri-ovulatory putrescine deficiency in older mice appears to adversely impact on oocyte maturation resulting in poor quality embryos (as assessed by blastocyst cell numbers) and early embryo death. This study demonstrates a natural and simple remedy to improve oocyte quality in older women. This study was supported by the NSERC, the March of Dimes Foundation, and the National Natural Science Foundation of China. The authors declare no competing interest.

Maternal DHA supplementation during pregnancy and offspring blood pressure at age 5y: follow‐up of a randomized controlled trial in Mexico (804.21)

Docosahexaenoic acid (DHA) supplementation in adults results in blood pressure improvements. The influence of prenatal DHA supplementation on blood pressure (BP) in childhood remains relatively unexplored. We investigated the effect of DHA supplementation during pregnancy on offspring blood pressure at age 5 y. In a double‐blind, randomized, controlled trial conducted in Mexico (POSGRAD), pregnant women were supplemented daily with 400 mg DHA or placebo from 18‐22 wk gestation to parturition. In the present follow up study, the blood pressure of 802 offspring children at age 5 years (82.4% of the birth cohort) was measured, using an automated blood pressure monitor (OMRON 711). Systolic (SBP) and diastolic blood pressure (DBP) were recorded as the average of three readings (SBP; mean +SD 93.1 + 8.2mmHg and DBP; 56.7 + 6.4mmHg).Between group differences in mean BP (DHA ‐ placebo) were: SBP, 0.56 (95% CI ‐0.50, 1.61) mmHg; and DBP, 0.15 (95% CI ‐0.72, 1.02) mmHg, after adjustment by side of the arm in which the measurement was performed, age at follow‐up, gender, child height and overweight status at age 5y, and maternal overweight at recruitment. Despite improvements in DHA status at birth, this study shows that prenatal DHA supplementation did not affect offspring blood pressure in early childhood.Grant Funding Source: Supported by NIH (HD058818; HDO43099), March of Dimes Foundation and CONACYT no.SALUD‐2008‐01‐87121

Born too soon: care before and between pregnancy to prevent preterm births: from evidence to action.

Providing care to adolescent girls and women before and between pregnancies improves their own health and wellbeing, as well as pregnancy and newborn outcomes, and can also reduce the rates of preterm birth. This paper has reviewed the evidence based interventions and services for preventing preterm births; reported the findings from research priority exercise; and prescribed actions for taking this call further. Certain factors in the preconception period have been shown to increase the risk for prematurity and, therefore, preconception care services for all women of reproductive age should address these risk factors through preventing adolescent pregnancy, preventing unintended pregnancies, promoting optimal birth spacing, optimizing pre-pregnancy weight and nutritional status (including a folic acid containing multivitamin supplement, and ensuring that all adolescent girls have received complete vaccination. Preconception care must also address risk factors that may be applicable to only some women. These include screening for and management of chronic diseases, especially diabetes; sexually-transmitted infections; tobacco and smoke exposure; mental health disorders, notably depression; and intimate partner violence. The approach to research in preconception care to prevent preterm births should include a cycle of development and delivery research that evaluates how best to scale up coverage of existing, evidence-based interventions, epidemiologic research that assesses the impact of implementing these interventions, and discovery science that better elucidates the complex causal pathway of preterm birth and helps to develop new screening and intervention tools. In addition to research, policy and financial investment is crucial to increasing opportunities to implement preconception care, and rates of prematurity should be included as a tracking indicator in global and national maternal child health assessments.DeclarationThis article is part of a supplement jointly funded by Save the Children's Saving Newborn Lives programme through a grant from The Bill & Melinda Gates Foundation and March of Dimes Foundation and published in collaboration with the World Health Organization (WHO). The original article was published in PDF format in the WHO Report "Born Too Soon: the global action report on preterm birth (ISBN 978 92 4 150343 30). The article has been reformatted for journal publication and has undergone peer review according to Reproductive Health's standard process for supplements and may feature some variations in content when compared to the original report. This co-publication makes the article available to the community in a full-text format.

Born too soon: preterm birth matters.

Urgent action is needed to address preterm birth given that the first country-level estimates show that globally 15 million babies are born too soon and rates are increasing in most countries with reliable time trend data. As the first in a supplement entitled "Born Too Soon", this paper focuses on the global policy context. Preterm birth is critical for progress on Millennium Development Goal 4 (MDG) for child survival by 2015 and beyond, and gives added value to maternal health (MDG 5) investments also linking to non-communicable diseases. For preterm babies who survive, the additional burden of prematurity-related disability may affect families and health systems. Prematurity is an explicit priority in many high-income settings; however, more attention is needed especially in low- and middle-income countries where the invisibility of preterm birth as well as its myths and misconceptions have slowed action on prevention and care. Recent global attention to preterm birth hit a tipping point in 2012, with the May 2 publication of Born Too Soon: The Global Action Report on Preterm Birth and with the 2nd annual World Prematurity Day on November 17 which mobilised the actions of partners in many countries to address preterm birth and newborn health. Interventions to strengthen preterm birth prevention and care span the continuum of care for reproductive, maternal, newborn and child health. Both prevention of preterm birth and implementation of care of premature babies require more research, as well as more policy attention and programmatic investment.DeclarationThis article is part of a supplement jointly funded by Save the Children's Saving Newborn Lives programme through a grant from The Bill & Melinda Gates Foundation and March of Dimes Foundation and published in collaboration with the World Health Organization (WHO). The original article was published in PDF format in the WHO Report "Born Too Soon: the global action report on preterm birth (ISBN 978 92 4 150343 30). The article has been reformatted for journal publication and has undergone peer review according to Reproductive Health's standard process for supplements and may feature some variations in content when compared to the original report. This co-publication makes the article available to the community in a full-text format.

Born too soon: care during pregnancy and childbirth to reduce preterm deliveries and improve health outcomes of the preterm baby.

Pregnancy and childbirth represent a critical time period when a woman can be reached through a variety of mechanisms with interventions aimed at reducing her risk of a preterm birth and improving her health and the health of her unborn baby. These mechanisms include the range of services delivered during antenatal care for all pregnant women and women at high risk of preterm birth, services provided to manage preterm labour, and workplace, professional and other supportive policies that promote safe motherhood and universal access to care before, during and after pregnancy. The aim of this paper is to present the latest information about available interventions that can be delivered during pregnancy to reduce preterm birth rates and improve the health outcomes of the premature baby, and to identify data gaps. The paper also focuses on promising avenues of research on the pregnancy period that will contribute to a better understanding of the causes of preterm birth and ability to design interventions at the policy, health care system and community levels. At minimum, countries need to ensure equitable access to comprehensive antenatal care, quality childbirth services and emergency obstetric care. Antenatal care services should include screening for and management of women at high risk of preterm birth, screening for and treatment of infections, and nutritional support and counselling. Health workers need to be trained and equipped to provide effective and timely clinical management of women in preterm labour to improve the survival chances of the preterm baby. Implementation strategies must be developed to increase the uptake by providers of proven interventions such as antenatal corticosteroids and to reduce harmful practices such as non-medically indicated inductions of labour and caesarean births before 39 weeks of gestation. Behavioural and community-based interventions that can lead to reductions in smoking and violence against women need to be implemented in conjunction with antenatal care models that promote women's empowerment as a strategy for reducing preterm delivery. The global community needs to support more discovery research on normal and abnormal pregnancies to facilitate the development of preventive interventions for universal application. As new evidence is generated, resources need to be allocated to its translation into new and better screening and diagnostic tools, and other interventions aimed at saving maternal and newborn lives that can be brought to scale in all countries.DeclarationThis article is part of a supplement jointly funded by Save the Children's Saving Newborn Lives programme through a grant from The Bill & Melinda Gates Foundation and March of Dimes Foundation and published in collaboration with the Partnership for Maternal, Newborn and Child Health and the World Health Organization (WHO). The original article was published in PDF format in the WHO Report "Born Too Soon: the global action report on preterm birth" (ISBN 978 92 4 150343 30), which involved collaboration from more than 50 organizations. The article has been reformatted for journal publication and has undergone peer review according to Reproductive Health's standard process for supplements and may feature some variations in content when compared to the original report. This co-publication makes the article available to the community in a full-text format.

Born too soon: care for the preterm baby.

As part of a supplement entitled "Born Too Soon", this paper focuses on care of the preterm newborn. An estimated 15 million babies are born preterm, and the survival gap between those born in high and low income countries is widening, with one million deaths a year due to direct complications of preterm birth, and around one million more where preterm birth is a risk factor, especially amongst those who are also growth restricted. Most premature babies (>80%) are between 32 and 37 weeks of gestation, and many die needlessly for lack of simple care. We outline a series of packages of care that build on essential care for every newborn comprising support for immediate and exclusive breastfeeding, thermal care, and hygienic cord and skin care. For babies who do not breathe at birth, rapid neonatal resuscitation is crucial. Extra care for small babies, including Kangaroo Mother Care, and feeding support, can halve mortality in babies weighing <2000 g. Case management of newborns with signs of infection, safe oxygen management and supportive care for those with respiratory complications, and care for those with significant jaundice are all critical, and are especially dependent on competent nursing care. Neonatal intensive care units in high income settings are de-intensifying care, for example increasing use of continuous positive airway pressure (CPAP) and this makes comprehensive preterm care more transferable. For health systems in low and middle income settings with increasing facility births, district hospitals are the key frontier for improving obstetric and neonatal care, and some large scale programmes now include specific newborn care strategies. However there are still around 50 million births outside facilities, hence home visits for mothers and newborns, as well as women's groups are crucial for reaching these families, often the poorest. A fundamental challenge is improving programmatic tracking data for coverage and quality, and measuring disability-free survival. The power of parent's voices has been important in high-income countries in bringing attention to preterm newborns, but is still missing from the most affected countries.DeclarationThis article is part of a supplement jointly funded by Save the Children's Saving Newborn Lives programme through a grant from The Bill & Melinda Gates Foundation and March of Dimes Foundation and published in collaboration with the Partnership for Maternal, Newborn and Child Health and the World Health Organization (WHO). The original article was published in PDF format in the WHO Report "Born Too Soon: the global action report on preterm birth" (ISBN 978 92 4 150343 30), which involved collaboration from more than 50 organizations. The article has been reformatted for journal publication and has undergone peer review according to Reproductive Health's standard process for supplements and may feature some variations in content when compared to the original report. This co-publication makes the article available to the community in a full-text format.

Inflammation biomarkers in vaginal fluid and preterm delivery.

Which inflammation biomarkers detected in the vaginal fluid are most informative for identifying preterm delivery (PTD) risk? Elevated interleukin (IL)-6 at mid-trimester was associated with increased odds of spontaneous PTD at <35 weeks and with PTD plus histologic chorioamnionitis (HCA), and had the greatest sensitivity for detecting these two PTD subtypes. Maternal and/or fetal inflammation play a role in some preterm deliveries, therefore inflammation biomarkers might help to identify women at greater risk. We examined 1115 women from the Pregnancy Outcomes and Community Health Study, a cohort study conducted from September 1998 through June 2004, for whom data were available on mid-pregnancy inflammatory biomarkers. At enrollment at 16-27 weeks gestation, vaginal fluid samples were collected from a swab and 15 eluted biomarkers were measured using the Meso Scale Discovery multiplex electrochemiluminescence platform. Associations of biomarkers with PTD were examined, according to clinical circumstance, week at delivery and presence/absence of HCA. Weighted logistic regression was used to determine odds ratios (OR) and 95% confidence intervals (CI) adjusted for race. Sensitivity and specificity were compared between individual and multiple biomarkers, identified by a bootstrapping method. Elevated IL-6 (>75th percentile) displayed the strongest association with spontaneous PTD <35 weeks (OR 2.3; CI 1.3-4.0) and PTD with HCA (OR 2.8; CI 1.4-6.0). The sensitivity of IL-6 to detect spontaneous PTD <35 weeks or PTD with HCA was 0.43 and 0.51, respectively, while specificity was 0.74 and 0.75, respectively. IL-6 plus IL1β, IL-6r, tumor necrosis factor-alpha or granulocyte-macrophage colony-stimulating factor increased specificity (range 0.84-0.88), but decreased sensitivity (range 0.28-0.34) to detect both PTD subtypes. Results were similar when a combination of IL-6 and bacterial vaginosis (BV) was explored. Thus, the use of multiple biomarkers did not detect PTD subtypes with a greater sensitivity than IL-6 alone, and IL-6 is a specific but non-sensitive marker for the detection of spontaneous PTD. Our ability to find small effect size associations between PTD and inflammation biomarkers (OR <2.0) might have been limited by the modest number of less common PTD subtypes in our population (e.g. spontaneous delivery <35 weeks, PTD accompanied by HCA) and by relatively higher variability for some cytokines, for example tumor necrosis factor-α, IL-12p70, IL-10 and granulocyte-macrophage colony-stimulating factor, that are less stable and commonly undetectable or detectable at low levels in human vaginal secretions. Larger studies are needed to further explore a role of inflammation biomarkers in combination with other risk factors, including specific BV-associated organisms, for the prediction of PTD subtypes. This work was supported by the National Institute of Child Health and Human Development, National Institute of Nursing, March of Dimes Foundation, Thrasher Research Foundation and Centers for Disease Control and Prevention. The authors have no conflicts of interest.

Retinoid Acid Specifies Neuronal Identity through Graded Expression of Ascl1

SummaryCell diversity and organization in the neural tube depend on the integration of extrinsic signals acting along orthogonal axes. These are believed to specify distinct cellular identities by triggering all-or-none changes in expression of combinations of transcription factors [1]. Under the influence of a common dorsoventral signal, sonic hedgehog, and distinct anterior-posterior (A-P) inductive signals [2, 3], two topographically related progenitor pools that share a common transcriptional code produce serotonergic and V3 neurons in the hindbrain and spinal cord, respectively [4–7]. These neurons have different physiological properties, functions, and connectivity [8, 9]. Serotonergic involvement in neuropsychiatric diseases has prompted greater characterization of their postmitotic repertoire of fate determinants, which include Gata2, Lmx1b, and Pet1 [10], whereas V3 neurons express Sim1 [4]. How distinct serotonergic and V3 neuronal identities emerge from progenitors that share a common transcriptional code is not understood. Here, we show that changes in retinoid activity in these two progenitor pools determine their fates. Retinoids, via Notch signaling, control the expression level in progenitors of the transcription factor Ascl1, which selects serotonergic and V3 neuronal identities in a dose-dependent manner. Therefore, quantitative differences in the expression of a single component of a transcriptional code can select distinct cell fates.

Is Cell-Free Fetal DNA From Maternal Blood Finally Ready for Prime Time?

See related article on page 890. Dr. Simpson is from the Florida International University, Miami, FL, and is Senior Vice President of Research and Global Programs with the March of Dimes Foundation, White Plains, NY; e-mail: [email protected]. Financial Disclosure Dr. Simpson has been an advisor to Bio Dx, Novartis, and Bayer.

Growth Factor Signaling Pathways in Lung Development and Cancer

Lung mesenchyme is a critical determinant of the shape and size of the lung, the extent and patterning of epithelial branching, the formation of the pulmonary vasculature, and the formation of mesenchymal components of the adult lung. Fibroblast Growth Factor 9 (FGF9) is expressed in developing lung epithelium and mesothelium and has an essential primary role in regulating mesenchymal growth and differentiation through signaling to mesenchymal FGF receptors (FGFRs) 1 and 2.Pleuropulmonary Blastoma (PPB) is a pediatric lung cancer syndrome with onset in late gestation and early childhood. PPB is a unique lung cancer in that its cellular origins are derived from lung mesenchyme. Germline loss‐of‐function mutations in Dicer1, an essential gene for the production of microRNAs, predisposes to PPB in humans. In this talk I will discus mechanisms by which FGF9 regulates mesenchymal growth and differentiation, evidence that Fgf9 may be regulated by microRNAs during lung development, and evidence that FGF9 may be an important factor for the initiation PPB.This work is funded by a grant from the March of Dimes foundation.

Challenges in defining and classifying the preterm birth syndrome

In 2009, the Global Alliance to Prevent Prematurity and Stillbirth Conference charged the authors to propose a new comprehensive, consistent, and uniform classification system for preterm birth. This first article reviews issues related to measurement of gestational age, clinical vs etiologic phenotypes, inclusion vs exclusion of multifetal and stillborn infants, and separation vs combination of pathways to preterm birth. The second article proposes answers to the questions raised here, and the third demonstrates how the proposed system might work in practice.

Fetal Loss of Adrenomedullin Causes Maternal Vascular Pathology Typical of Preeclampsia.

Adrenomedullin (AM) is a 52-amino acid peptide vasodilator that is elevated 3 to 5-fold in normal human pregnancies but is often blunted in pregnancy complications such as fetal growth restriction, gestational diabetes, and preeclampsia. Our previous studies have shown that genetic reduction of maternal AM leads to poor pregnancy outcomes, but the role of fetal AM remains unknown. Therefore, we examined placentas of Adm-/- and Adm+/+ pups at midgestation for structural, functional, and developmental phenotypes. Histologic and morphometric analysis revealed normal differentiation and layer formation in the Adm-/- placentas. Doppler ultrasound of umbilical cord blood flow showed that placental impedance was unaffected by fetal loss of Adm. Although histology and lectin staining showed no difference in the total cross-sectional area of fetal and maternal vessels, the fetal vessels of the Adm-/- placentas were large and underbranched compared to those of wildtype littermates. Electron microscopy of methyl-methacrylate casts confirmed that Adm-/- placentas had reduced fetal vessel branching; a hallmark feature of preeclampsia. Strikingly, the Adm-/- placentas also had reduced numbers of decidual natural killer (NK) cells and reduced remodeling of maternal spiral arteries; other known features of preeclampsia. To further investigate the effects of AM signaling on NK cells, isolated primary NK cells were treated with AM, which resulted in significantly altered cytokine expression compared to untreated controls. Primary smooth muscle cell cultures exposed to supernatant from AM-treated NK cells also showed a significant increase in smooth muscle cell apoptosis, indicating that the AM-mediated change in NK cell cytokine expression may be required for normal spiral artery remodeling. To confirm that this phenotype is specific to lack of fetal AM, ovarian transplantation was performed from Adm+/- to Adm+/+ females. The placentas of Adm-/- pups from wildtype mothers receiving heterozygote ovary transplants recapitulated the reduced NK cell invasion and failed spiral artery remodeling originally seen in heterozygous mothers, demonstrating that lack of AM specifically from the fetal compartment is responsible for the observed preeclampsia-like pathology. Furthermore, we were able to rescue this phenotype by generating a transgenic mouse model with increased fetal AM expression, termed Admhi/hi. High fetal AM led to recruitment of increased decidual NK cells and normal spiral artery remodeling. RT-PCR screening revealed that numerous cytokines known to play important roles in the placenta had opposite regulation in Admhi/hi versus Adm-/- placentas. Taken together, these data indicate that loss of fetal AM causes reduced fetal vascularization and maternal spiral artery remodeling characteristic of preeclampsia, and that these phenotypes can be rescued by overexpressing fetal AM. These findings demonstrate that the dosage of fetal AM is a critical factor for adapting the maternal vasculature to pregnancy and may be important in preventing the development of preeclampsia. This work is supported by The Burroughs Wellcome Fund, NIH/NICHD RO1 HD46970, HD060860, NIH/NHLBI F30 HL104778, and the March of Dimes Foundation. (poster)

Effect of prenatal supplementation with docosahexanoic acid on infant development: a randomized placebo‐controlled trial in Mexico.

The omega‐3 fatty acid docosahexanoic acid (DHA) is deposited in the brain early in life and plays an important role in neural and visual function. Little is known however about the benefits of ingesting DHA supplements during pregnancy for infant development. We conducted a randomized double‐blind placebo‐controlled trial of 400 mg/day algal DHA (Martek) from week 20 of gestation through delivery (n=1094 pregnant women) and followed the offspring (n=973 live births). Child development was measured in 730 children (75% of birth cohort) using the Spanish version of the Bayley Scales of Infant Development (BSID)‐II at 18 mo of age; we calculated standardized psychomotor development (PDI) and mental development indices (MDI). PDI and MDI scores (mean ± S.D.) were 94.3 ± 10.7 and 93.0 ± 8.9 in the DHA group (n=365) and 95.2 ± 9.3 and 93.3 ± 9.8 in the placebo group (n=365), respectively. Intent to treat differences in PDI and MDI scores at 18 mo were 0.90 (95% CI: −0.55, 2.35; p=0.23) and 0.26 (95% CI: −1.10, 1.63; p=0.70), respectively. Prenatal supplementation with 400 mg algal DHA did not result in improved global development scores at 18 mo of age in a study population (Mexico) with low intakes of DHA. Follow‐up of the cohort is ongoing. Funding: NIH (HD043099) and March of Dimes Foundation.

126 Adipose tissue engineering by human adipose-derived stromal cell seeded gelatin sponges

The large bulk of lost craniofacial soft tissue is mainly adipose tissue. Current surgical approaches following tumor resection and trauma have limitations. Adipose derived-stromal cells (ADSCs), also named as preadipocytes, have been demonstrated to be a good cell candidate for adipose tissue engineering. However, characteristics of scaffold played important role in a successful tissue engineered adipose tissue. Objectives: The present study is to investigate whether in vitro adipogenesis could be induced both in a monolayer and 3-D by ADSCs. Methods: Healthy adult ADSCs were isolated by liposuction from fat pads. After discarding the unattached cells, the ADSCs were cultured in DMEM supplemented with 10% FBS and 1% antibiotics until they reached confluence. After trypsinization, ADSCs were subcultured at a density of 105 cells/100 mm-dish as a first passage. The monolayer and 3-D adipogenesis were assessed after seeding 105 ADSCs into each well of 6-well plates and 5 × 106 ADSCs/ml into biodegradable collagen sponges, respectively, followed by replacement of basic medium with adipogenic one. Results: In both monolayer and 3-D scaffolds, lipids containing adipocytes were observed 1 week after adipogenic stimulation, and further increased over the time. By contrast, no adipogenesis was observed in ADSCs cultured in basic medium. Conclusions: Human adipose-derived stromal cells isolated from fat pads can readily be induced toward adipogenesis in vitro. The in vivo adipogenesis potential of ADSCs is being investigated. (Supported by Whitaker Biomedical Engineering Foundation, March of Dimes Foundation)